Articles de revues : « Moi-pro »

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Cardona-Mendoza, Andrés, Nelly Stella Roa Molina, Diana Marcela Castillo, Gloria Inés Lafaurie et Diego Fernando Gualtero Escobar. « Human Coronary Artery Endothelial Cell Response to Porphyromonas gingivalis W83 in a Collagen Three-Dimensional Culture Model ». Microorganisms 12, n^o 2 (24 janvier 2024) : 248. http://dx.doi.org/10.3390/microorganisms12020248.

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P. gingivalis has been reported to be an endothelial cell inflammatory response inducer that can lead to endothelial dysfunction processes related to atherosclerosis; however, these studies have been carried out in vitro in cell culture models on two-dimensional (2D) plastic surfaces that do not simulate the natural environment where pathology develops. This work aimed to evaluate the pro-inflammatory response of human coronary artery endothelial cells (HCAECs) to P. gingivalis in a 3D cell culture model compared with a 2D cell culture. HCAECs were cultured for 7 days on type I collagen matrices in both cultures and were stimulated at an MOI of 1 or 100 with live P. gingivalis W83 for 24 h. The expression of the genes COX-2, eNOS, and vWF and the levels of the pro-inflammatory cytokines thromboxane A2 (TXA-2) and prostaglandin I2 (PGI2) were evaluated. P. gingivalis W83 in the 2D cell culture increased IL-8 levels at MOI 100 and decreased MCP-1 levels at both MOI 100 and MOI 1. In contrast, the 3D cell culture induced an increased gene expression of COX-2 at both MOIs and reduced MCP-1 levels at MOI 100, whereas the gene expression of eNOS, vWF, and IL-8 and the levels of TXA2 and PGI2 showed no significant changes. These data suggest that in the collagen 3D culture model, P. gingivalis W83 induces a weak endothelial inflammatory response.

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Militzer, Burkhard, et William B. Hubbard. « Relation of Gravity, Winds, and the Moment of Inertia of Jupiter and Saturn ». Planetary Science Journal 4, n^o 5 (1 mai 2023) : 95. http://dx.doi.org/10.3847/psj/acd2cd.

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Abstract We study the relationship of zonal gravity coefficients, J 2n , zonal winds, and axial moment of inertia (MoI) by constructing models for the interiors of giant planets. We employ the nonperturbative concentric Maclaurin spheroid method to construct both physical (realistic equation of state and barotropes) and abstract (small number of constant-density spheroids) interior models. We find that accurate gravity measurements of Jupiter’s and Saturn’s J 2, J 4, and J 6 by the Juno and Cassini spacecraft do not uniquely determine the MoI of either planet but do constrain it to better than 1%. Zonal winds (or differential rotation (DR)) then emerge as the leading source of uncertainty. For Saturn they are predicted to decrease the MoI by 0.4% because they reach a depth of ∼9000 km, while on Jupiter they appear to reach only ∼3000 km. We thus predict DR to affect Jupiter’s MoI by only 0.01%, too small by one order of magnitude to be detectable by the Juno spacecraft. We find that winds primarily affect the MoI indirectly via the gravity harmonic J 6, while direct contributions are much smaller because the effects of pro- and retrograde winds cancel. DR contributes +6% and −0.8% to Saturn’s and Jupiter’s J 6 value, respectively. This changes the J 6 contribution that comes from the uniformly rotating bulk of the planet that correlates most strongly with the predicted MoI. With our physical models, we predict Jupiter’s MoI to be 0.26393 ± 0.00001. For Saturn, we predict 0.2181 ± 0.0002, assuming a rotation period of 10:33:34 hr that matches the observed polar radius.

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Rice, J. Hunter, Margaret M. McDaniel, Alyson Holland et Shigetoshi Eda. « Modelling Bovine Granuloma Formation In Vitro upon Infection with Mycobacterium Avium Subspecies Paratuberculosis ». Veterinary Sciences 6, n^o 4 (12 octobre 2019) : 80. http://dx.doi.org/10.3390/vetsci6040080.

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Mycobacterium avium subspecies paratuberculosis (Map) causes chronic granulomatous disease in cattle and ruminant livestock, causing substantial economic losses. Current vaccines delay clinical signs but cannot train the immune system to fully eradicate latent Map. During latency, Map uses host defenses, cage-like macrophage clusters called granuloma, as incubators for months or years. We used an in vitro model to investigate the early coordination of macrophages into granuloma upon Map infection over ten days. We found that at multiplicities of infection (MOI; Map:macrophages) of 1:2 and below, the macrophages readily form clusters and evolve pro-inflammatory cytokines in keeping with a cell-mediated immune response. At higher MOIs, viability of host macrophages is negatively impacted. At 1:4 MOI, we quantified viable Map in our model and confirmed that intracellular Map reproduced over the first five days of infection. Host cells expressed Type 1-specific cytokines, and Map-infected macrophages displayed reduced motility compared to Map-exposed, uninfected macrophages, suggesting an important role for uninfected macrophages in the early aggregative response. Reported is the first in vitro JD granuloma model capturing Map and macrophage viability, size distribution of resulting clusters, motility of monocyte-derived macrophages, and cytokine response during clustering, allowing quantitative analysis of multiple parameters of the Map-specific granulomatous response.

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Maldonado, David, Vivian Ouyang, Jade Owens, Andrew Jimenez, Benjamin Saks, Payam Sabetian, Ajay Lall et Benjamin Domb. « Paper 57 : Labral Tear Management in Patients Aged 40 Years and Older Undergoing Primary Hip Arthroscopy : A Propensity-Matched Case-Control Study Comparing Labral Reconstruction to Labral Repair with Minimum Two-Year Follow-Up. *ACCEPTED TO AJSM* ». Orthopaedic Journal of Sports Medicine 10, n^o 7_suppl5 (1 juillet 2022) : 2325967121S0062. http://dx.doi.org/10.1177/2325967121s00620.

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Objectives: To report minimum two-year patient-reported outcome (PRO) scores, survivorship, and secondary surgeries, in patients aged ≥ 40 years who underwent primary hip arthroscopy with labral reconstruction compared to a propensity-matched primary labral repair group. Methods: Data were prospectively collected and retrospectively reviewed for patients who underwent a primary hip arthroscopy for femoroacetabular impingement syndrome from January 2014 to June 2018. Patients aged ≥ 40 years who underwent a labral reconstruction or a labral repair and had preoperative and minimum two-year PROs for the modified Harris Hip Score (mHHS), Non-arthritic Hip Score (NAHS), and visual analog scale (VAS) for pain were included. Patients with prior ipsilateral hip conditions and surgery, Tonnis grade > 1, hip dysplasia, or worker’s compensation status were excluded. Patients in the reconstruction group were propensity-matched 1:2 to patients in the repair group based on age, sex, and body mass index. Secondary surgeries and the achievement of the minimal clinically importance difference (MCID), patient acceptable symptomatic state (PASS), and maximum outcome improvement (MOI) were recorded. Results: Fifty-three and 106 hips were included in the labral reconstruction and repair groups, respectively. The average follow-up time was 37.6 months. The average age for the reconstruction and repair groups were 48.01 ± 5.4 years and 48.61 ± 6.0 years, respectively. Both groups achieved significant improvements in all PROs at minimum two-years with similar achievements of MCID, PASS, and MOI. Both groups showed comparable secondary surgeries rates. Conclusions: Patients aged 40 years and older who received primary labral repair and primary labral reconstruction, achieved similar significant improvements in all PROs, VAS pain, and patient satisfaction at minimum two-year follow-up, with comparable rates of secondary surgeries and achieving MCID, PASS, and MOI. Based on these findings, labral repair remains the gold standard treatment for viable labrum in this population group, while reconstruction is a useful alternative for irreparable labrum.

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Maldonado, David R., Vivian W. Ouyang, Jade S. Owens, Andrew E. Jimenez, Benjamin R. Saks, Payam W. Sabetian, Ajay C. Lall et Benjamin G. Domb. « Labral Tear Management in Patients Aged 40 Years and Older Undergoing Primary Hip Arthroscopy : A Propensity-Matched Case-Control Study With Minimum 2-Year Follow-up ». American Journal of Sports Medicine 49, n^o 14 (15 octobre 2021) : 3925–36. http://dx.doi.org/10.1177/03635465211046915.

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Background: Previous literature has suggested that primary acetabular labral reconstruction leads to lower secondary surgery rates than does labral repair for patients aged ≥40 years. Purpose: To report minimum 2-year patient-reported outcome (PRO) scores, survivorship, and secondary surgeries in patients aged ≥40 years who underwent primary hip arthroscopy with labral reconstruction compared with a propensity-matched primary labral repair group. Study Design: Case-control study; Level of evidence, 3. Methods: Data were prospectively collected and retrospectively reviewed for patients who underwent a primary hip arthroscopy for femoroacetabular impingement syndrome between January 2014 and June 2018. Patients aged ≥40 years who underwent a labral reconstruction or a labral repair and had preoperative and minimum 2-year PROs for the modified Harris Hip Score, Nonarthritic Hip Score, and visual analog scale (VAS) for pain were included. Patients with previous ipsilateral hip conditions and surgery, Tönnis grade >1, hip dysplasia, or workers’ compensation status were excluded. Patients in the reconstruction group were propensity matched 1:2 to patients in the repair group based on age, sex, and body mass index. Secondary surgeries and achievement of the minimal clinically important difference (MCID), patient acceptable symptom state (PASS), and maximum outcome improvement (MOI) were recorded. Results: A total of 53 and 106 hips were included in the labral reconstruction and repair groups, respectively. The average follow-up time was 37.6 months. The average ages for the reconstruction and repair groups were 48.01 ± 5.4 years and 48.61 ± 6.0 years, respectively. Both groups achieved significant improvements in all PROs at a minimum of 2 years, with similar achievements of MCID, PASS, and MOI, and comparable secondary surgery rates. Conclusion: Patients aged ≥40 years who received primary labral repair and primary labral reconstruction achieved similar significant improvements in all PROs, VAS pain, and patient satisfaction at the minimum 2-year follow-up, with comparable rates of secondary surgeries and achieving MCID, PASS, and MOI. Based on these findings, labral repair remains the gold standard treatment for viable labrum in this population group, while reconstruction is a useful alternative for irreparable labrum.

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L Katche, Musong, Augustine B Makokha, Siagi O Zachary et Muyiwa S Adaramola. « Solar Photovoltaic Systems : A Technical and Economic Feasibility Design Approach with Homer Pro ». International Journal of Advances in Scientific Research and Engineering 09, n^o 11 (2023) : 21–30. http://dx.doi.org/10.31695/ijasre.2023.9.11.2.

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With the declining cost of solar photovoltaic (PV) modules and batteries used for energy storage, many users are now shifting towards solar energy because of its renewable nature and availability. Though this is a great step to combat climate change, most of the solar systems installed always fail due to poor system sizing. For this reason, optimal systems are required to be deployed. This paper presents a technical and economic feasibility design approach for a solar PV system using Hybrid Optimization of Multiple Energy Resources (HOMER) Pro software. The design is based on site-specific data collected from Moi University in Kenya. The temperature and solar radiation data were collected fromthe weather station of the university, while the power demand data was collected from the Margaret Thatcher Library of the university using the PCE-360 power analyzer.The simulation results show that a 100kW solar PV system is required to power the Margaret Thatcher library with a financial investment of KES 32,000,000. This system is strongly recommended to be used by the university as it will ensure reliability of power supply for students to study and also save on costs incurred on utility bills.

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Bajaber, Gamar, Constantine Akwanalo et Shameem Ali. « Proportion, Precipitators and Prognostic Marker of 30-Day Heart Failure Readmission at MOI Teaching and Referral Hospital, Eldoret, Kenya ». International Journal of Research and Scientific Innovation VII, n^o XI (2023) : 110–25. http://dx.doi.org/10.51244/ijrsi.2023.1011008.

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Background: Heart failure (HF) affects 26 million people globally. It is associated with a high 30-day readmission rate due to multiple cardiovascular and non-cardiovascular precipitants. Readmissions are associated with high mortality, which may be predicted by pre-discharge N-terminal pro brain natriuretic peptide (NT-proBNP). There is no data on HF readmission rates, its precipitators and use of NT-proBNP as a prognostic marker at Moi Teaching and Referral Hospital (MTRH) in Western Kenya. Objective: To determine the 30-day proportion of HF readmission, the precipitators of HF and the association between NT-proBNP and 30 days’ readmission. Methods:This was a six-month prospective cohort study where we carried out a census and recruited adult participants admitted with HF at Moi Teaching and Referral Hospital. At discharge from hospital, an interviewer administered questionnaire was administered and blood samples for NT-proBNP drawn. Upon readmission, precipitators for HF were identified and compliance to therapy assessed using European heart failure Self Care Behavioral Scale. Continuous variables were summarized using median and IQR, categorical variables using frequencies and percentages. Associations were determined using Chi square, Fishers exact and Wilcoxon rank test. A p-value of <0.05 was considered statistically significant. Results: From April to November 2018, 94 participants were recruited into the study; with median age 48 years (IQR 31,70), 58 (62%) were female, 35 (38%) consumed alcohol and 25(25%) smoked. Hypertension was the commonest comorbidity 20 (21%) while cardiomyopathy was the underlying etiology for HF in 58 (63%). HF with reduced Ejection Fraction (HFrEF) was present in 76% of the participants while 24% had HF with preserved Ejection Fraction (HFpEF). Sixty percent of the participants had NT-proBNP levels of >4137 pg/ml, which implied poor prognosis. Of 17 readmitted patients, 12 participants (12.8%) were readmitted within 30 days at MTRH. Six (6.4%) participants were lost to follow up. Median time to readmission was 14 days, (IQR 7, 26). Pneumonia (55%) was the commonest precipitator of HF readmissions, followed by arrhythmias (atrial fibrillation) in 5 (42%), anemia 4 (33%), noncompliance 4 (33%), acute kidney injury 2 (16.6%) and no identified precipitator 1 (8.3%). There was no association between NT-proBNP and readmission (p =0.584) or NT-pro BNP and survival (p=0.773). Readmission was associated with a high mortality (p=0.008) with 50% of readmitted participants dying during the readmission period. The total mortality of both readmitted and non-readmitted participants was 16% at the end of the 6 months’ study period. Conclusions:In this cohort of participants with HF the proportion of 30 days readmission was high. Infections, mainly pneumonia was the commonest precipitator of readmission. Discharge NT-proBNP did not predict likelihood of 30 days’ readmission. Mortality was higher among participants readmitted within 30 days. Recommendations: Measures like pneumoccal vaccinations should be implemented to prevent pneumonia. Early appointments to cardiac clinic (less than 2 weeks post discharge) should be given to screen for precipitators and reduce HF readmissions. A follow up study to assess % change in NT pro BNP in relation to 30 days readmission.

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A . Hussein, Ahmed, et Adnan F. Hassan. « Performance Enhancement of Luminescent Solar Concentrator by using Mixing Fluorescent Colors and Nanoparticles ». Journal of Kufa-Physics 14, n^o 01 (12 août 2022) : 17–25. http://dx.doi.org/10.31257/2018/jkp/2022/140106.

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Liquid zinc acetate nanoparticles were added to a fluorescent organic dye (fluorescein sodium) to prepare three concentrations (2 × 10-5, 7×10-5, 1×10-4) moI/L, and the absorbance of the dye was measured before and after adding Liquid nanomaterial by using two devices (SCINCO Mega-2100 UV/visible Spectrophotometer.) as well as measuring the flux of the dye before and after adding zinc acetate nanoparticles using the (Spectrofluorometer F96 PRO). The Stoke displacement (Δη), the radiative age (τfm), the fluorescence lifetime (τf), and the quantitative efficiency (Qfm) were calculated. Absorption and fluorescence curves were drawn using the Excel program. The MATLAB program was also used to measure the area under the absorption and fluorescence spectra curves. It was found that the fluorescence intensity of fluorescein sodium dye has increased compared to the intensity of fluorescence before adding the nanomaterial and thus increasing the quantitative efficiency, which in turn helps in improving the performance of the photovoltaic concentrator, resulting in an improvement in the efficiency of the solar cell.

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Barletta, Raul G., John P. Bannantine, Judith R. Stabel, Ezhumalai Muthukrishnan, Dirk K. Anderson, Enakshy Dutta, Vamsi Manthena, Mostafa Hanafy et Denise K. Zinniel. « Mycobacterium avium subsp. paratuberculosis Candidate Vaccine Strains Are Pro-apoptotic in RAW 264.7 Murine Macrophages ». Vaccines 11, n^o 6 (10 juin 2023) : 1085. http://dx.doi.org/10.3390/vaccines11061085.

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Mycobacterium avium subsp. paratuberculosis (MAP) is the etiological agent of Johne’s disease, a severe gastroenteritis of ruminants. This study developed a model cell culture system to rapidly screen MAP mutants with vaccine potential for apoptosis. Two wild-type strains, a transposon mutant, and two deletion mutant MAP strains (MOI of 10 with 1.2 × 106 CFU) were tested in murine RAW 264.7 macrophages to determine if they induce apoptosis and/or necrosis. Both deletion mutants were previously shown to be attenuated and immunogenic in primary bovine macrophages. All strains had similar growth rates, but cell morphology indicated that both deletion mutants were elongated with cell wall bulging. Cell death kinetics were followed by a real-time cellular assay to measure luminescence (apoptosis) and fluorescence (necrosis). A 6 h infection period was the appropriate time to assess apoptosis that was followed by secondary necrosis. Apoptosis was also quantified via DAPI-stained nuclear morphology and validated via flow cytometry. The combined analysis confirmed the hypothesis that candidate vaccine deletion mutants are pro-apoptotic in RAW 264.7 cells. In conclusion, the increased apoptosis seen in the deletion mutants correlates with the attenuated phenotype and immunogenicity observed in bovine macrophages, a property associated with good vaccine candidates.

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Knezevic, Petar, Aleksandra Petrovic Fabijan, Damir Gavric, Jovana Pejic, Zsolt Doffkay et Gábor Rakhely. « Phages from Genus Bruynoghevirus and Phage Therapy : Pseudomonas Phage Delta Case ». Viruses 13, n^o 10 (30 septembre 2021) : 1965. http://dx.doi.org/10.3390/v13101965.

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The applicability and safety of bacteriophage Delta as a potential anti-Pseudomonas aeruginosa agent belonging to genus Bruynoghevirus (family Podoviridae) was characterised. Phage Delta belongs to the species Pseudomonas virus PaP3, which has been described as a temperate, with cos sites at the end of the genome. The phage Delta possesses a genome of 45,970 bp that encodes tRNA for proline (Pro), aspartic acid (Asp) and asparagine (Asn) and does not encode any known protein involved in lysogeny formation or persistence. Analysis showed that phage Delta has 182 bp direct terminal repeats at the end of genome and lysogeny was confirmed, neither upon infection at low nor at high multiplicity of infection (MOI). The turbid plaques that appear on certain host lawns can result from bacteriophage insensitive mutants that occur with higher frequency (10−4). In silico analysis showed that the genome of Delta phage does not encode any known bacterial toxin or virulence factor, determinants of antibiotic resistance and known human allergens. Based on the broad host range and high lytic activity against planktonic and biofilm cells, phage Delta represents a promising candidate for phage therapy.

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Ochwoto, Missiani, Colins O. Oduma, Julius Oyugi, Dufton Mwaengo, Bartholomew N. Ondigo, James H. Kimotho, Alex K. Maiyo, Ruth M. Nyangacha, Gladys Chesumbai et Elijah Songok. « Human TP53 gene polymorphisms among patients with hepatocellular carcinoma and chronic hepatitis B in Kenya ». F1000Research 8 (6 août 2019) : 1364. http://dx.doi.org/10.12688/f1000research.19416.1.

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Background: Human TP53 is the gatekeeper for generation of human cells and is highly conserved. Any alteration/mutation to TP53 adversely affects the regulatory function of the protein, potentially resulting in cancer. This study investigated mutations in codons 7 and 249 of TP53, among patients with hepatocellular carcinoma (HCC) and chronic hepatitis B virus (HBV) infection at the Moi Teaching and Referral Hospital (MTRH), Eldoret, Kenya. Methods: In total, 33 HBV-positive patients attending MTRH hospital between September 2013 and July 2017 were purposely selected from medical records for the study; those with HCC were confirmed from the cancer registry. The patients were aged between 25-67 years, with a male-to-female ratio of 1.1:1. Blood samples were collected from the patients. DNA was extracted, amplified and sequenced using TP53 forward and reverse primers. Gene mutation detection and analysis was done on exons 4 and 7 Results: Of the 33 patients, 75.8% were chronically infected with HBV and had HCC; the rest were HBsAg positive without HCC. Homozygous proline was prevalent (54.5%) at exon 4 codon 72, followed by heterozygous Arg/Pro (33.3%) and lastly homozygous Arg/Arg (12.1%,). Pro/Pro allele was frequent in HCC group while Arg/Arg allele was common in patients without HCC. There was no significant association between the HCC and codon polymorphisms (p=0.12). In exon 7, codon 249, 24.2% of patients had an Arg-Ser mutation of which, 75.0% had HCC and 25.0% did not. There was no significant association between HCC patients and codon 249 mutation (p=0.15). Conclusion: TP53 is a gene gate keeper, the mutations under study may dependently play a role in HCC development. This study did not find any association or clear mutational pattern between P53 mutations and HCC development. Therefore, TP53 mutation is a poor indicator for prognosis and a tumor’s biological behavior among HBV-positive subjects in Kenya.

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Ochwoto, Missiani, Colins O. Oduma, Julius Oyugi, Dufton Mwaengo, Bartholomew N. Ondigo, James H. Kimotho, Alex K. Maiyo, Ruth M. Nyangacha, Gladys Chesumbai et Elijah Songok. « Human TP53 gene polymorphisms among patients with Hepatocellular carcinoma and chronic hepatitis B infection in Kenya ». F1000Research 8 (12 mars 2024) : 1364. http://dx.doi.org/10.12688/f1000research.19416.2.

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Background Human TP53 is the gatekeeper for generation of human cells and is highly conserved. Some alteration/mutation in TP53 adversely affects the regulatory function of the protein, potentially resulting in cancer. This study investigated mutations in codons 72 and 249 of TP53, among patients with hepatocellular carcinoma (HCC) and chronic hepatitis B virus (HBV) infection at the Moi Teaching and Referral Hospital (MTRH), Eldoret, Kenya. Methods In total, 33 HBV-positive patients attending MTRH hospital between September 2013 and July 2017 were purposely selected from medical records for the study; those with HCC were confirmed from the cancer registry. The patients were aged between 25-67 years, with a male-to-female ratio of 1.1:1. Blood samples were collected from the patients. DNA was extracted, amplified and sequenced using TP53 forward and reverse primers. Gene mutation detection and analysis was done on exons 4 codon 72 and exon 7 codon 249. Results Of the 33 patients, 75.8% were chronically infected with HBV and had HCC; the rest were HBsAg positive without HCC. Homozygous proline was prevalent (54.5%) at exon 4 codon 72, followed by heterozygous Arg/Pro (33.3%) and lastly homozygous Arg/Arg (12.1%). Pro/Pro allele was frequent in HCC group while Arg/Arg allele was common in patients without HCC. There was no significant association between the HCC and codon polymorphisms (P=0.12). In exon 7, codon 249, 24.2% of patients had an Arg/Ser mutation of which, 75.0% had HCC and 25.0% did not. There was no significant association between HCC patients and codon 249 mutation (P=0.15). Conclusion TP53 is a gene gate keeper, the mutations under study may dependently play a role in HCC development. This study did not find any association between TP53 mutations and presence of HCC. Therefore, TP53 Arg-72 and Ser-249 mutation is not a clear prognosis indicator for hepatocellular carcinoma among HBV infected patients in Kenya.

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Loeffler, Juergen, Markus Mezger et Hermann Einsele. « Mycophenolic Acid Causes Dysfunction of Human Polymorphonuclear Neutrophils and Dendritic Cells during the Interaction with Aspergillus fumigatus. » Blood 110, n^o 11 (16 novembre 2007) : 3849. http://dx.doi.org/10.1182/blood.v110.11.3849.3849.

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Abstract Immunosuppressive drugs are a variety of substances to prevent transplant rejection. There is increasing data that immunosuppressants not only modulate lymphocyte function but also interfere with other immune effector cells. In this study, we analyzed the influence of Mycophenolic acid (MPA) on the activity of human polymorphonuclear neutrophils (PMNs) and dendritic cells (DCs) during the interaction with the mold Aspergillus fumigatus. PMNs were obtained from blood using Bicoll separation. Release of reactive oxygen species (ROS) was detected photometrically by using dichlorofluorescein. Monocytes were isolated by magnetic-associated cell sorting followed by differentiation into DCs using GM-CSF and IL-4. DCs were either co-cultivated with MPA (10 μg/ml), with A. fumigatus germ tubes (MOI=1) or both stimuli. Whole-genome microarray analyses (Affymetrix U133A) or quantitative real-time PCR assays were performed to quantify differentially regulated genes. MPA could be identified as an enhancer of the A. fumigatus induced oxidative burst of PMNs while fungal killing efficiency was not affected. Incubation of fully differentiated Mo-derived DCs in the presence of MPA did not alter gene expression patterns. However, if DCs were treated during early differentiation processes, DC development was influenced by MPA resulting in increased apoptosis. In addition, we revealed impaired expression of a variety of pro-inflammatory immune response genes (TNF-α, CXCL10, IL-12). In conclusion, MPA influences PMN and DC function during immune defense against fungi. MPA triggers an acute inflammatory syndrome by enhancing the A. fumigatus induced oxidative burst. Furthermore, innate and adaptive immune responses might be impaired because of a reduced number of DCs and lower levels of pro-inflammatory cytokines and chemokines.

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Valverde, Araceli Maria, Samantha Schaller et Afsar Raza Naqvi. « MicroRNA and periodontal pathogen interaction in shaping innate immune response ». Journal of Immunology 206, n^o 1_Supplement (1 mai 2021) : 112.08. http://dx.doi.org/10.4049/jimmunol.206.supp.112.08.

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Abstract MicroRNA (miR) play a critical role in the pathogenesis of immune-mediated diseases, however, their role in periodontal disease (PD) remain poorly studied. We hypothesize that periopathogens (P. gingivalis [Pg] and A. actinomycemtemcomitans [Aa]) impair host immune response by perturbing miR expression. Naïve CD14+ monocytes were isolated from human PBMC and differentiated to M1 MΦ or M2 MΦ in presence of GM-CSF or M-CSF for 7 days. Cells were challenged with Pg or Aa (both 100 MOI) for 4 h and 24 h. Total RNA was isolated to quantitate expression of pro- and anti-inflammatory miRNAs, cell lysate was prepared for protein expression of NFκB, PU.1 and CEBP/β by western blot and supernatants were collected to analyze cytokine profiles. We observed that expression of pro-inflammatory miRNAs viz., miR-125a and miR-155a are increased in M1-MΦ and M2-MΦ challenged with Pg and Aa. Surprisingly, the expression of anti-inflammatory miR-146a was differentially expressed in both MΦ, but miR-511 expression was reduced. miR-142-3p expression in response to Pg and Aa challenge was decreased suggesting its possible anti-inflammatory function. To examine the impact of miR-142-3p on Pg and Aa-mediated immune responses, we examined PU.1 levels, a known TF regulating miR-142 expression, and noticed a significant decrease in PU.1 and it correlates with mature miR-142-3p levels. miR-142-3p transfected MΦ challenged with Pg or Aa exhibit altered secretion of inflammatory cytokines. In conclusion, periopathogen-mediated activation of NFκB and miR-155 augments inflammatory signaling by suppressing expression of anti-inflammatory miR-142-3p by decreasing PU.1 levels. Perturbation of miR by periodontal bacteria is central to the pathogenesis of PD.

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Babalghith, Ahmad O., Hayder M. Al-kuraishy, Ali I. Al-Gareeb, Michel De Waard, Jean-Marc Sabatier, Hebatallah M. Saad et Gaber El-Saber Batiha. « The Potential Role of Growth Differentiation Factor 15 in COVID-19 : A Corollary Subjective Effect or Not ? » Diagnostics 12, n^o 9 (24 août 2022) : 2051. http://dx.doi.org/10.3390/diagnostics12092051.

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Coronavirus disease 2019 (COVID-19) is primarily caused by various forms of severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) variants. COVID-19 is characterized by hyperinflammation, oxidative stress, multi-organ injury (MOI)-like acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). Different biomarkers are used in the assessment of COVID-19 severity including D-dimer, ferritin, lactate dehydrogenase (LDH), and hypoxia-inducible factor (HIF). Interestingly, growth differentiation factor 15 (GDF15) has recently become a potential biomarker correlated with the COVID-19 severity. Thus, this critical review aimed to determine the critical association between GDF15 and COVID-19. The perfect function of GDF15 remains not well-recognized; nevertheless, it plays a vital role in controlling cell growth, apoptosis and inflammatory activation. Furthermore, GDF15 may act as anti-inflammatory and pro-inflammatory signaling in diverse cardiovascular complications. Furthermore, the release of GDF15 is activated by various growth factors and cytokines including macrophage colony-stimulating factor (M-CSF), angiotensin II (AngII) and p53. Therefore, higher expression of GDF15 in COVID-19 might a compensatory mechanism to stabilize and counteract dysregulated inflammatory reactions. In conclusion, GDF15 is an anti-inflammatory cytokine that could be associated with the COVID-19 severity. Increased GDF15 could be a compensatory mechanism against hyperinflammation and exaggerated immune response in the COVID-19. Experimental, preclinical and large-scale clinical studies are warranted in this regard.

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Katche, Musong L., Augustine B. Makokha, Siagi O. Zachary et Muyiwa S. Adaramola. « Techno-Economic Assessment of Solar–Grid–Battery Hybrid Energy Systems for Grid-Connected University Campuses in Kenya ». Electricity 5, n^o 1 (29 janvier 2024) : 61–74. http://dx.doi.org/10.3390/electricity5010004.

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This paper presents the techno-economic feasibility of using grid-connected PV hybrid systems to supply power in large grid-dependent academic institutions. The study was conducted using the administration building of Moi University in Kenya. The power consumption profile of the building was collected using a PCE-360 power analyzer. The peak load demand was found to be 60 kW. Using random variability constants of 4% for day-to-day and 4% time-step load variability, a peak demand of 70.58 kW was obtained, which was used in our simulation. The solar radiation and temperature data for this site were collected from the weather station of the university. The hybrid system was simulated using HOMER Pro software. It was found from the simulation results that the optimal system was the solar PV/grid without battery storage, which had a levelized cost of energy (LCOE) of KSH 8.78/kWh (USD 0.072), net present cost (NPC) of KSH 27,974,492 (USD 230,813), capital expenditure (CAPEX) of KSH 26,300,000 (USD 216,997), and a simple payback period (SPBP) of 5.08 years for a 25-year life span. This system, when compared to the existing grid, showed an 83.94% reduction in the annual electricity bill of the administration building. These results demonstrate a reduction in energy cost by a renewable energy fraction of 67.1%.

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Snuggs, J., R. Senter, J. Whitt et C. Le Maitre. « PCRX-201, A NOVEL GENE THERAPY TREATMENT APPROACH FOR INTERVERTEBRAL DISC DEGENERATION ». Orthopaedic Proceedings 105-B, SUPP_9 (17 avril 2023) : 91. http://dx.doi.org/10.1302/1358-992x.2023.9.091.

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Low back pain affects 80% of the population with half of cases attributed to intervertebral disc (IVD) degeneration. However, the majority of treatments focus on pain management, with none targeting the underlying pathophysiological causes. PCRX-201 presents a novel gene therapy approach that addresses this issue. PCRX-201 codes for interleukin-1 receptor antagonist (IL-1Ra), the natural inhibitor of the pro-inflammatory cytokine IL-1, which orchestrates the catabolic degeneration of the IVD. Our objective here is to determine the ability of PCRX-201 to infect human nucleus pulposus (NP) cells and tissue to increase the production of IL-1Ra and assess downstream effects on catabolic protein production.Degenerate human NP cells and tissue explants were infected with PCRX-201 at 0 or 3000 multiplicities of infection (MOI) and subsequently cultured for 5 days in monolayer (n=7), 21 days in alginate beads (n=6) and 14 days in tissue explants (n=5). Cell culture supernatant was collected throughout culture duration and downstream targets associated with pain and degeneration were assessed using ELISA.IL-1Ra production was increased in NP cells and tissue infected with PCRX-201. The production of downstream catabolic proteins such as IL-1β, IL-6, MMP3, ADAMTS4 and VEGF was decreased in both 3D-cultured NP cells and tissue explants.Here, we have demonstrated that a novel gene therapy, PCRX-201, is able to infect and increase the production of IL-1Ra in degenerate NP cells and tissue in vitro. The increase of IL-1Ra also resulted in a decrease in the production of a number of pro-inflammatory and catabolic proteins, suggesting PCRX-201 enables the inhibition of IL-1-driven IVD degeneration. At present, no treatments for IVD degeneration target the underlying pathology. The ability of FX201 to elicit anti-catabolic responses is promising and warrants further investigation in vitro and in vivo, to determine the efficacy of this exciting, novel gene therapy.

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Fox, Sarah, Kieran Ryan, Soumita Das, Shiela Crowe et Peter Ernst. « C1q promotes recognition and engulfment of H. pylori infected AGS cells and modulates inflammatory cytokine production in macrophages. (P4019) ». Journal of Immunology 190, n^o 1_Supplement (1 mai 2013) : 131.10. http://dx.doi.org/10.4049/jimmunol.190.supp.131.10.

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Abstract H pylori resides in the gastric lumen and is a significant risk factor for gastritis and gastric cancer. This organism promotes apoptosis of gastric epithelial cells that are subsequently cleared by local phagocytes. C1q, the recognition component of the complement cascade, is an important molecule in immune regulation where it promotes uptake of apoptotic cells and modulates inflammation. We demonstrated that H pylori infection induces apoptosis of gastric AGS cells in vitro and that apoptotic epithelial cells are recognized and engulfed by macrophages independent of the method of cell death. Further, C1q binds to apoptotic cells and is important in the interaction between apoptotic cells and macrophages. We show that the absence of serum reduced attachment of AGS cells to phagocytes by up to 90% and the addition of 40μg/ml of C1q was sufficient to restore normal binding. A comparable effect of C1q on internalization of AGS cells was observed using a flow cytometric assay. C1q, in the absence of serum, attenuated IL-6, TNF-α and IL-1β production induced by 100 ng/ml of LPS or 100 MOI H. pylori compared to controls (45-60% reductions). In conclusion, we show that C1q is important in the resolution of inflammation by facilitating the engulfment of apoptotic cells and inhibiting the pro-inflammatory cytokine responses. Further, the inhibition of host responses attributed to the engulfment of apoptotic cells may be induced by extracellular factors including C1q.

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Shymialevich, Dziyana, Stanisław Błażejak, Paulina Średnicka, Hanna Cieślak, Agnieszka Ostrowska, Barbara Sokołowska et Michał Wójcicki. « Biological Characterization and Genomic Analysis of Three Novel Serratia- and Enterobacter-Specific Virulent Phages ». International Journal of Molecular Sciences 25, n^o 11 (29 mai 2024) : 5944. http://dx.doi.org/10.3390/ijms25115944.

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Due to the high microbiological contamination of raw food materials and the increase in the incidence of multidrug-resistant bacteria, new methods of ensuring microbiological food safety are being sought. One solution may be to use bacteriophages (so-called phages) as natural bacterial enemies. Therefore, the aim of this study was the biological and genomic characterization of three newly isolated Serratia- and Enterobacter-specific virulent bacteriophages as potential candidates for food biocontrol. Serratia phage KKP_3708 (vB_Sli-IAFB_3708), Serratia phage KKP_3709 (vB_Sma-IAFB_3709), and Enterobacter phage KKP_3711 (vB_Ecl-IAFB_3711) were isolated from municipal sewage against Serratia liquefaciens strain KKP 3654, Serratia marcescens strain KKP 3687, and Enterobacter cloacae strain KKP 3684, respectively. The effect of phage addition at different multiplicity of infection (MOI) rates on the growth kinetics of the bacterial hosts was determined using a Bioscreen C Pro growth analyzer. The phages retained high activity in a wide temperature range (from −20 °C to 60 °C) and active acidity values (pH from 3 to 12). Based on transmission electron microscopy (TEM) imaging and whole-genome sequencing (WGS), the isolated bacteriophages belong to the tailed bacteriophages from the Caudoviricetes class. Genomic analysis revealed that the phages have linear double-stranded DNA of size 40,461 bp (Serratia phage KKP_3708), 67,890 bp (Serratia phage KKP_3709), and 113,711 bp (Enterobacter phage KKP_3711). No virulence, toxins, or antibiotic resistance genes were detected in the phage genomes. The lack of lysogenic markers indicates that all three bacteriophages may be potential candidates for food biocontrol.

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Gaultier, Gaël. « Moi, jeune et privé d’emploi ». Projet 361, n^o 6 (2017) : 5. http://dx.doi.org/10.3917/pro.361.0005.

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Rawls, Amy S., Jill R. Woloszynek et Daniel C. Link. « Lentiviral-Mediated RNAi Knockdown of SBDS in Murine Hematopoietic Progenitors Impairs Their Engraftment Potential and Granulocytic Differentiation. » Blood 106, n^o 11 (16 novembre 2005) : 635. http://dx.doi.org/10.1182/blood.v106.11.635.635.

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Abstract Shwachman-Bodian Diamond Syndrome (SDS) is an autosomal recessive disorder characterized by congenital neutropenia, pancreatic exocrine insufficiency and bony abnormalities. About 90% of SDS patients have mutations in the novel SBDS gene. The SBDS protein is conserved from archaea to mammals; however, the cellular functions of SBDS, as well as its contribution to the molecular pathogenesis of this disease, are not yet known. In this study, we developed an RNAi knockdown model to investigate the consequences of a reduction in SBDS protein on hematopoiesis. Three lentiviral RNAi constructs directed against SBDS were generated and tested in BAF3 cells, a murine pro-B cell line. SBDS protein reduction of 78 ± 11%, 63 ± 14%, and 27 ± 6% was observed. The most efficient lentiviral SBDS RNAi construct was used to transduce primary murine hematopoietic cells, with knockdown similar to that observed in BAF3 cells. In addition, empty lentiviral and Cathepsin G (CG) RNAi lentiviral constructs were studied to control for non-specific effects of lentivirus infection and/or induction of the RNAi machinery. In each case, the lentiviral vector contained a yellow fluorescent protein (YFP) expression cassette to track transduced cells. Murine hematopoietic progenitors (Kit+, Lin-) were transduced with these lentiviral vectors and transplanted into irradiated syngeneic recipients. Initially, a high multiplicity of infection (MOI) was used, resulting in a transduction efficiency of ~75%. Interestingly, 5/5 animals transplanted with SBDS RNAi transduced cells died by 19 days post-transplantation due to engraftment failure, compared to 0/5 animals transplanted with empty lentivirus transduced cells. To study the long-term consequence of SBDS knockdown, cells were transduced at a lower MOI, resulting in a transduction efficiency of ~15%. Engraftment of SBDS RNAi cells (N=8) was modestly (3-fold), yet consistently, reduced compared to both empty lentivirus (N=8) and CG RNAi (N=5) controls. However, once engraftment of SBDS RNAi cells was established, the percentage of transduced (YFP+) cells remained stable for at least 3 months. Moreover, there was no evidence of lineage skewing, as a similar percentage of YFP+ cells was observed in all hematopoietic lineages. Importantly, the SBDS knockdown effect was maintained in YFP+ cells recovered from mice 3 months post-transplantation (~70% reduction in SBDS expression). Neutropenia is the most common hematopoietic abnormality in patients with SDS. Thus, we also characterized the effect of reduced SBDS function on granulocytic differentiation. Specifically, the ability of SBDS RNAi transduced Kit+, Lin- progenitors to generate hematopoietic colonies and differentiate into mature granulocytes was assessed. No defect in the ability of transduced progenitors to form CFU-GM or CFU-G was observed. However, a modest, yet significant, delay in neutrophil maturation was observed. Mature neutrophils constitute only 34.6 ± 7.3% of SBDS RNAi granulocyte cultures by day 7, compared to 59.7 ± 6.9% and 60.1 ± 12.9% in empty lentivirus and CG RNAi cultures, respectively. In summary, we established an RNAi model for SDS and have shown that a 70% reduction in SBDS protein levels in hematopoietic progenitors is sufficient for (i) impaired engraftment upon bone marrow transplantation and (ii) modest delay in neutrophil maturation in vitro. These findings provide the first experimental evidence that reduced SBDS function may drive the hematopoietic defects seen in SDS patients.

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Herzog, Philippe. « Plus social que moi, tu meurs ». Projet 284, n^o 1 (2005) : 42. http://dx.doi.org/10.3917/pro.284.0042.

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Stavrakaki, Eftychia, Wouter B. L. van den Bossche, Rutger Balvers, Lisette B. Vogelezang, Cristina Teodosio, Dana A. M. Mustafa, Willem de Koning et al. « EXTH-54. CO-CULTURE OF PATIENT-DERIVED GLIOBLASTOMA CELL LINES WITH AUTOLOGOUS PBMCS FOR THE INVESTIGATION OF ONCOLYTIC VIRUS RESPONSE ». Neuro-Oncology 24, Supplement_7 (1 novembre 2022) : vii221. http://dx.doi.org/10.1093/neuonc/noac209.852.

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Abstract BACKGROUND The dismal prognosis of glioblastoma (GBM) patients, with a median survival of less than 15 months despite maximal therapy urgently warrants new therapeutic approaches. Clinical trials employing oncolytic viruses (OVs) have shown encouraging results, however, in each OV clinical trial only a small subset of patients responded to treatment. As inter-tumoral heterogeneity has been the key challenge in treating GBM, we hypothesized that development of an in vitro co-culture model for assessment of viral replication and subsequent immune response might ultimately predict in vivo OV efficacy for individual GBM patients. METHODS 20 patient-derived GBM cell cultures were tested with dose-ranges of two clinically relevant OVs (DNX2401 and rQnestin34.5 V1) to determine the EC50 values (half-maximal effective concentration). To discriminate between responders and non-responders six of these cultures were infected using MOI 10 for DNX2401 and 0.25 for rQnestin34.5V1, and co-cultured with autologous PBMCs. OV-induced changes in gene and protein expression of immune associated genes were assessed using targeted gene expression (NanoString Technology) and ELISA. RESULTS DNX2401 EC50 values ranged from 0.35 to > 600 and from 0.04 to 1.77 for rQnestin34.5V1. Induction of pro-inflammatory cytokines and chemokines differed per virus and per patient. Enhanced OV infection or oncolysis efficiency did not lead to increased levels of IFNγ production. Importantly, longer exposure to dexamethasone prior to PBMC isolation correlated with suppressed IFNγ production. CONCLUSION We established an autologous GBM cells/PBMCs co-culture model which reflects inter-tumoral heterogeneity in terms of cytokine induction and virus-specific changes in gene and protein expression upon OV infection. Immune activation is not directly related to degree of infection or oncolysis and can be hampered by prolonged prior dexamethasone use. These first results support our hypothesis that improved prediction of response rates in oncolytic virotherapy for GBM may require a personalized approach with an in vitro test system.

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Allazkani, Nour, et Martin Monti-Lalaubie. « « L’engagement m’a appris à être moi-même » ». Revue Projet N°373, n^o 6 (2019) : 5. http://dx.doi.org/10.3917/pro.373.0005.

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Kumar, Ashish, Quanzhi Li, Wendy A. Hudson, Thien N. Sam, Weili Chen et John H. Kersey. « Evi1 Promotes Leukemogenesis by Anti-Apoptotic Rather Than Differentiation-Blocking Effects in Murine MLL-AF9 Leukemia ». Blood 112, n^o 11 (16 novembre 2008) : 593. http://dx.doi.org/10.1182/blood.v112.11.593.593.

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Abstract In a search for molecular targets in MLL -leukemias, we found that transformed hematopoietic stem cells in knock-in MLL-AF9 mice expressed high levels of Evi1 compared to their wild type counterparts (Chen et al, Cancer Cell, 2008). About 50% of human AMLs with MLL rearrangements also express Evi1 and Evi1-expressing human leukemias are associated with an aggressive disease. The current experiments were designed to study mechanisms in which Evi1 is important for the pathogenesis of leukemia. To investigate its role in MLL-AF9 leukemia, we studied the effect of Evi1 knock-down in a cell line derived from a leukemic MLL-AF9 knock-in mouse. This cell line (4166) displays a myelo-monocytic phenotype, similar to human MLL-AF9 leukemias and induces leukemia with high efficiency when transplanted in wild type mice. To knock down Evi1, we screened multiple shRNA constructs cloned into a lentivirus expression vector (The RNAi Consortium/Open Biosystems). Of five shRNAs screened, two clones (E95 and E97) consistently inhibited cell growth in a dose dependent manner at multiplicities of infection (MOI) of 10 or greater. In one representative experiment, at 5 days post transduction at an MOI of 100, growth of E95 transduced 4166 cells was decreased by >80% compared to control-virus transduced cells. Evi1 mRNA was decreased in E95 transduced cells by >70% compared to controls at 48 hours. When cultured in semi-solid methylcellulose media for 7 days, E95 transduced cells formed significantly fewer colonies than control virus transduced cells (457.8 ± 24.7 and 847.8 ± 92.9 colonies per 1000 cells respectively, p<0.02). Surprisingly however, there was no change in the proportions of dense vs. loose colonies indicating that loss of Evi1 did not result in differentiation. Morphologic analysis also showed no change in morphology of E95 transduced cells. Further confirming a lack of differentiation-inhibiting effects of Evi1, quantitative RT-PCR assays showed no change in expression of genes associated with myeloid differentiation (integrin alpha-L, integrin beta-5, lysozyme, Csf-1). In Evi1 knock-down cells, analysis of DNA content by flow cytometry at 48 hours showed no change in the proportions of cells in the G1/G0, S and G2/M phases of the cell cycle. In contrast to lack of an effect on differentiation, there was an increase in apoptosis with Evi1 knock-down as evidenced by an increase in the proportion of cells stained with Annexin V and Propidium iodide (51% vs. 29%) or a pan activated-caspase-marker and Propidium iodide (41% vs. 15%, E95 vs. control virus transduced cells respectively). To investigate the role of Evi1 in leukemia in vivo, 105 cells transduced with either E95 or control virus were injected into irradiated wild type mice. Establishment of leukemia was confirmed by leukocytosis, splenomegaly and evidence of malignant infiltrates in the spleen by morphology and FACS. Eight of nine mice that received control virus transduced cells died of leukemia by day 170 while all those given E95 transduced cells were alive. Leukemia development was significantly delayed in the animals receiving E95 transduced cells (p<0.001, log rank test) with 44% of the animals being long term survivors. Overall, our results suggest that contrary to the view that leukemic-oncogenes induce self-renewal coupled to a block in differentiation, in murine MLL-AF9 leukemia Evi1 inhibits pro-apoptotic pathways and promotes cell-survival without a differentiation-block. Additionally, given the critical requirement of this transcription factor in the survival of leukemia cells, targeting Evi1 may have therapeutic potential to treat leukemia.

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Dugast, César, et Alexia Soyeux. « Les entreprises, l’État, le climat et moi : chacun sa part ». Revue Projet N°375, n^o 2 (2020) : 38. http://dx.doi.org/10.3917/pro.375.0038.

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Vettraino, Jean. « Jade Lindgaard, JE CRISE CLIMATIQUE. La planète, ma chaudière et moi ». Projet 343, n^o 6 (2014) : 93. http://dx.doi.org/10.3917/pro.343.0093.

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Salafutdinov, Ilnur I., Dilara Z. Gatina, Мaria I. Markelova, Ekaterina E. Garanina, Sergey Yu Malanin, Ilnaz M. Gazizov, Svetlana Khaiboullina, Albert A. Rizvanov et Rustem I. Islamov. « Transcriptomic Landscape of Umbilical Cord Blood Mononuclear Cells after Genetic Modification ». Blood 136, Supplement 1 (5 novembre 2020) : 33–34. http://dx.doi.org/10.1182/blood-2020-138664.

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Introduction: Therapeutic application of umbilical cord blood mononuclear cells (UCB-MCs) has been actively studied for at least during the last 30 years. Currently, UCB-MCs are extensively being investigated in the regeneration of the central nervous system (CNS) and the treatment of human neurodegenerative disorders. Translational studies dedicated to the application of UCB-MCs have revealed their capacity for stimulation of neurogenesis in the aged brain and promising potency for being therapeutic agents for treating such diseases as Alzheimer`s disease, amyotrophic lateral sclerosis (ALS), ischemic stroke, traumatic brain injury, Parkinson`s disease etc. Still the exact mechanism providing therapeutic effect remains unclear. Several studies modulating neurodegeneration have demonstrated immunomodulating and pro-inflammatory activity of UCB-MCs. Moreover, the unique feature of UCB-MCs, which underlies in the optional concordance of HLA or immunosuppression during transplantation, has been shown. At the same time, the main challenge in the application of UCB-MCs is the limited amount of available cells from a single donor. Therefore, the development of methods increasing therapeutic potency is an important task when using UCB-MC. In this regard, in our opinion, one of the options for increasing the therapeutic potential of cells can be ex vivo genetic modification of the transplanted UCB-MC. This approach is beneficial to get cells with the desired therapeutic properties. At the same time, little is known about the effect of genetic modification and over-expression of therapeutic genes on the UCB-MC transcriptome. In this regard in our present study, we evaluated the transcriptomic landscape of gene engineered UCB-MC. Methods: UCB-MCs were separated using Ficoll density gradient and modified recombinant adenoviruses (Ad-EGFP or Ad-VEGF165, MOI 10). Total RNA from all test samples was extracted using Trizol reagent. The quality and concentration of the isolated RNA samples were evaluated using Agilent Bioanalyzer 2100. Total mRNA from genetically modified and non-treated cells was sequenced Sequencing-By-Synthesis (SBS) technology on Illumina NextSeq 500 platform in 2×75 bp mode. After quality control, reads were aligned to human reference transcriptome GRCh38 using Kallisto pseudoaligner. Differentially expressed transcripts and genes were calculated with R package "sleuth". Results: We comprehensively profiled the whole-transcriptome landscape of human genetically modified UCB-MC. Totally 12 cDNA libraries, obtained from 6 individual donors, were analyzed. Transcriptomic analysis has revealed 2,4-2,8×106 of paired reads. A total of 10164 genes in the RNA-seq data were detected and analyzed. UCB-MCs were shown to express a broad range of pro- and anti-inflammatory cytokines, chemokines, growth factors, and metalloproteinases. The principal component analysis (PCA) of the RNA-seq data showed that samples representing different biological conditions do not differ from each other and are grouped according to the source of their receipt (isolation) (Fig.1A). Genetic modification and expression of transgenes did not lead to a global shift in the transcriptome profile of UCB-MC. At the same time, the recombinant genes EGFP (log2FC = 7.15, q <0.05) and VEGF (log2FC = 4.41, q <0.05), as expected, showed increased expression compared to NTC. We performed Gene Ontology (GO) analysis of the expressed genes. The results demonstrated that most genes associated with biological processes were related to metabolism. In the category of cellular components, most detected genes were associated with cellular membrane and nucleus, while in molecular function category - protein binding (Fig.1B). Conclusion: It has been shown that genetic modification and expression of UCB-MC transgenes did not affect the global transcriptome profile. Transcriptome profiling can be useful in the creation and testing of personalized gene cell products that meet biological safety and efficacy criteria. This work was supported by the RFBR grant №18-44-160029 and subsidy allocated to Kazan Federal University for the state assignment in the sphere of scientific activities 0671-2020-0058. Disclosures No relevant conflicts of interest to declare.

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Castro-Rea, Julián. « LA NATION, C’EST MOI : THE ENCOUNTER OF QUÉBEC AND ABORIGINAL NATIONALISMS ». Constitutional Forum / Forum constitutionnel 13 (26 juillet 2011) : 2005. http://dx.doi.org/10.21991/c9t674.

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Proponents of Québec’s independence justify their goal with the claim that their province is the cradle of one of Canada’s “Founding Nations.” In so doing, they bypass the self-perception of the dominant, politicized Aboriginal peoples, who perceive themselves as forming the “First Nations” of what is now Canada. These contending views, neither of which is yet constitutionally fully recognized, are bound to clash whenever issues of self-government are raised within Québec’s boundaries. Such a situation arose at the time of the 1995 Québec referendum on sovereignty, which was met with adamant opposition from Aboriginal groups, especially the Cree, the Inuit and the Mohawk. In reaction, Québec’s pro-independence government at the time accused Aboriginal peoples of being Ottawa’s instrument, and repeated the debatable argument that Québec has maintained the most respectful policy towards Aboriginal peoples among all Canadian provinces.

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Soares, Rodrigo Pedro, Igor Campos Fontes, Felipe Dutra-Rêgo, Jeronimo Nunes Rugani, Paulo Otávio L. Moreira, Vânia Lúcia Ribeiro da Matta, Gabriela Venícia Araujo Flores et al. « Unveiling the Enigmatic nature of six neglected Amazonian Leishmania (Viannia) species using the hamster model : Virulence, Histopathology and prospection of LRV1 ». PLOS Neglected Tropical Diseases 18, n^o 8 (9 août 2024) : e0012333. http://dx.doi.org/10.1371/journal.pntd.0012333.

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American tegumentary leishmaniasis (ATL) is highly endemic in the Amazon basin and occurs in all South American countries, except Chile and Uruguay. Most Brazilian ATL cases are due to Leishmania (Viannia) braziliensis, however other neglected Amazonian species are being increasingly reported. They belong to the subgenus L. (Viannia) and information on suitable models to understand immunopathology are scarce. Here, we explored the use of the golden hamster Mesocricetus auratus and its macrophages as a model for L. (Viannia) species. We also studied the interaction of parasite glycoconjugates (LPGs and GIPLs) in murine macrophages. The following strains were used: L. (V.) braziliensis (MHOM/BR/2001/BA788), L. (V.) guyanensis (MHOM/BR/85/M9945), L. (V.) shawi (MHOM/BR/96/M15789), L. (V.) lindenbergi (MHOM/BR/98/M15733) and L. (V.) naiffi (MDAS/BR/79/M5533). In vivo infections were initiated by injecting parasites into the footpad and were followed up at 20- and 40-days PI. Parasites were mixed with salivary gland extract (SGE) from wild-captured Nyssomyia neivai prior to in vivo infections. Animals were euthanized for histopathological evaluation of the footpads, spleen, and liver. The parasite burden was evaluated in the skin and draining lymph nodes. In vitro infections used resident peritoneal macrophages and THP-1 monocytes infected with all species using a MOI (1:10). For biochemical studies, glycoconjugates (LPGs and GIPLs) were extracted, purified, and biochemically characterized using fluorophore-assisted carbohydrate electrophoresis (FACE). They were functionally evaluated after incubation with macrophages from C57BL/6 mice and knockouts (TLR2-/- and TLR4-/-) for nitric oxide (NO) and cytokine/chemokine production. All species, except L. (V.) guyanensis, failed to generate evident macroscopic lesions 40 days PI. The L. (V.) guyanensis lesions were swollen but did not ulcerate and microscopically were characterized by an intense inflammatory exudate. Despite the fact the other species did not produce visible skin lesions there was no or mild pro-inflammatory infiltration at the inoculation site and parasites survived in the hamster skin/lymph nodes and even visceralized. Although none of the species caused severe disease in the hamster, they differentially infected peritoneal macrophages in vitro. LPGs and GIPLs were able to differentially trigger NO and cytokine production via TLR2/TLR4 and TLR4, respectively. The presence of a sidechain in L. (V.) lainsoni LPG (type II) may be responsible for its higher proinflammatory activity. After Principal Component analyses using all phenotypic features, the clustering of L. (V.) lainsoni was separated from all the other L. (Viannia) species. We conclude that M. auratus was a suitable in vivo model for at least four dermotropic L. (Viannia) species. However, in vitro studies using peritoneal cells are a suitable alternative for understanding interactions of the six L. (Viannia) species used here. LRV1 presence was found in L. (V.) guyanensis and L. (V.) shawi with no apparent correlation with virulence in vitro and in vivo. Finally, parasite glycoconjugates were able to functionally trigger various innate immune responses in murine macrophages via TLRs consistent with their inflammatory profile in vivo.

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Yu, Lei, Greggory Myers, Beth McGee, Xiaofang Liu, Gallagher Patrick, Richard King, Ann Friedman et al. « Whole-Genome CRISPR-Cas9 Screen Identifies ZBTB7A As a Potential Therapeutic Target for Cda-II ». Blood 142, Supplement 1 (28 novembre 2023) : 12. http://dx.doi.org/10.1182/blood-2023-182246.

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Congenital dyserythropoietic anemia type II (CDA-II) is an autosomal recessive disease characterized by anemia, ineffective erythropoiesis, and increased bone marrow bi-nucleated erythroblasts. CDA-II is caused by loss-of-function mutations in SEC23B, which encodes a component of coat protein complex II (COP-II) vesicles/tubules that transport secretory proteins from the endoplasmic reticulum to the Golgi apparatus. Mammals express two SEC23 paralogs, SEC23A and SEC23B. We have previously shown that SEC23A is functionally interchangeable with SEC23B and that increased expression of SEC23A rescues the SEC23B-deficient erythroid differentiation defect observed in CDA-II. Here, we utilized our recently generated clonal SEC23B deficient HUDEP-2 cell line to identify novel therapeutic targets for CDAII. SEC23B deficient HUDEP2 cells survive and grow normally when cultured as pro-erythroblasts in ‘maintenance media’. However, upon culturing these cells in ‘differentiation media’, which results in semi-synchronous erythroid differentiation, differentiated SEC23B-null HUDEP2 cells exhibit reduced growth, impaired differentiation, increased bi-nucleated erythroid cells, and erythroid cell death, features of CDA-II. To identify novel genes that play important roles in the pathophysiology of CDAII, we developed a functional genome-scale CRISPR knockout screen to define genes that when deleted, rescue the SEC23B-null erythroid differentiation defect. SEC23B null HUDEP2 cells cultured in maintenance media were infected with the hGeCKO-v2 lentiviral library at a multiplicity of infection (MOI) of ~0.3, allowing most transduced cells to receive 1 sgRNA (and Cas9) to knockout 1 gene only. Transduced cells were puromycin selected and passaged in expansion media for 14 days. SEC23B-null HUDEP2 cells were then differentiated for 10 days. Cells were harvested prior to differentiation (Day 0, D0), and differentiated (orthochromatic) cells were sorted at D10 of differentiation. Integrated sgRNAs were quantified by deep sequencing. sgRNAs targeting ZBTB7A were amongst the most enriched in D10 compared to D0 erythroid cells. To validate these findings, we transduced SEC23B-null HUDEP2 cells with two independent and efficient ZBTB7A-targeting sgRNAs. We found that ZBTB7A deletion (using either of the 2 sgRNAs) rescued the lethality and differentiation defect of SEC23B null HUDEP2 cells undergoing differentiation. We next generated 7 clonal cell lines with combined deletion for ZBTB7A and SEC23B and confirmed that these cells exhibited normal growth and differentiation indistinguishable from wildtype HUDEP2 cells. The rescuing effect of targeting ZBTB7A was also validated in SEC23B mutated human CD34+ hematopoietic stem and progenitor cell culture. Since SEC23A can functionally replace SEC23B in erythroid cells, we quantified the SEC23A mRNA level in SEC23B-null HUDEP2 cells deleted for ZBTB7A, by qRT-PCR. In early preliminary results, deletion of ZBTB7A resulted in a profound (~10 fold) increase in SEC23A mRNA expression, to a level predicted to be sufficient to rescue the SEC23B null erythroid differentiation defect. ChIP-seq analysis demonstrated that ZBTB7A occupies the SEC23A promoter in HUDEP2 cells. Taken together, these data suggest that ZBTB7A represses SEC23A expression during erythroid maturation and that in the setting of ZBTB7A deletion, SEC23A expression is de-repressed, resulting in rescue of the CDA-II erythroid differentiation defect. Genome editing of the ZBTB7A binding sites in the SEC23A promoter is ongoing in SEC23B-null HUDEP-2 cells. We will determine the impact of the latter editing on SEC23A expression and erythroid differentiation.

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Franca Peri Giglio. « Modelo Educativo sustentado en Valores Espirituales para la vida, la autorrealización y trascendencia ». GACETA DE PEDAGOGÍA, n^o 48 (23 janvier 2024) : 17–30. http://dx.doi.org/10.56219/rgp.vi48.2438.

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La espiritualidad es la cualidad propia de la naturaleza trascendente del ser humano que define su sentido y propósito de vida. La educación juega un rol fundamental en este mecanismo de autoco-nocimiento y autorrealización ya que su finalidad no es sólo educare -instruir, entrenar- sino también educere -sacar a la luz, hacer salir-. Al respecto, la presente investigación desarrolló un modelo que operacionaliza lo espiritual a través de valores centrados en el autoconocimiento y la emancipación del Ser, e incorpora a los actores del hecho educativo como participantes activos del proceso en el marco de la educación permanente. El diseño se nutrió de referencias exitosas en el mundo donde las prácticas pedagógicas desarrollan cualidades que promue-ven la integridad y autonomía. Se concluyó que esta propuesta podrá validarse en tanto evidencie transformación interna y externa de los involucrados, en las instituciones y espacios de la sociedad de la cual forman parte. ABSTRACT Spirituality is the quality proper to the transcendent nature of human beings that defines their meaning and purpose in life. Education plays a fundamental role in this mechanism of self-know-ledge and self-realization since its purpose is not only educare -to instruct, to train- but also educere -to bring to light, to bring out-. In this regard, the present research developed a model that operationalizes the spiritual through values centered on self-know-ledge and emancipation of the Self, and incorporates the actors of the educational process as active participants in the process within the framework of lifelong education. The design was nourished by successful references in the world where pedagogical practices develop qualities that promote integrity and autonomy. It was concluded that this proposal can be validated as long as it evidences internal and external transformation of those involved, in the institutions and spaces of the society of which they are part. Key words: Self-realization, Self-knowledge, Spiritual Values, Educational Models, Pedagogy of Being, Continuing Education RESUMO A espiritualidade é a qualidade da natureza transcendente dos seres humanos que define seu significado e propósito na vida. A educação dessempenha um papel fundamental nesse mecanismo de autoconhecimento e autorrealização, uma vez que seu propósito não é apenas educare - instruir, treinar - mas também educere - trazer à luz, trazer à tona. A esse respeito, a presente pesquisa desenvolveu um modelo que operacionaliza o espiritual por meio de valores centrados no autoconhecimento e na emancipação do eu, e incorpora os atores do processo educacional como participantes ativos do processo dentro da estrutura da aprendizagem ao longo da vida. O projeto foi alimentado por referências bem-sucedidas no mundo em que as práticas pedagógicas desenvolvem qualidades que promovem a integridade e a auto-nomia. Concluiu-se que essa proposta pode ser validada desde que evidencie a transformação interna e externa dos emvolvidos, nas instituições e nos espaços da socie-dade da qual fazem parte. Palavras-chave: Autorrealização, Autoconhecimento, Valores Espirituais, Modelos Educacionais, Pedagogia do Eu, Educação ao Longo da Vida RÉSUMÉ La spiritualité est la qualité de la nature transcendante des êtres humains qui définit le sens et le but de leur vie. L'éducation joue un rôle fondamental dans ce mécanisme de conna-issance et de réalisation de soi, puisque son but n'est pas seulement educare - instruire, former - mais aussi educere - mettre en lumière, faire ressortir. À cet égard, la présente recherche a développé un modèle qui opérationnalise la spiritualité à travers des valeurs centrées sur la connaissance de soi et l'émancipation du moi, et qui intègre les acteurs du processus éducatif en tant que participants actifs au processus dans le cadre de l'apprentissage tout au long de la vie. La com-ception a été nourrie par des références réussies dans le monde où les pratiques pédagogiques développent des qualités qui promeuvent l'intégrité et l'autonomie. Il a été conclu que cette pro-position peut être validée tant qu'elle témoigne d'une transformation interne et externe des personnes impliquées, dans les institutions et les espaces de la société dont elles font partie. Mots-clés: Réalisation de soi, connaissance de soi, valeurs spirituelles, modèles éducatifs, pédagogie du soi, éducation tout au long de la vie.

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Merkushova, E. D., E. M. Khasanova et L. V. Gankovskaya. « Mechanisms of innate immunity in pathogenesis of psoriasis : approaches to targeted therapy ». Medical Immunology (Russia) 22, n^o 3 (21 mai 2020) : 449–58. http://dx.doi.org/10.15789/1563-0625-moi-1949.

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Psoriasis is a chronic auto-inflammatory, genetically determined dermatosis, being multifactorial by origin, characterized by hyperproliferation of epidermis, affected keratinocyte differentiation and inflammatory reaction in dermis. The disease is characterized by a tendency to spread over the area of lesion, and involvement of articular tissue in the pathological process, which significantly affects the living standards of patients and causes their disability. There are many provoking factors that contribute to occurrence of psoriasis, or progression of existing psoriatic process in individuals with a genetic predisposition. These factors include adverse climatic conditions, skin trauma, exposure to ultraviolet light, burns, infections, etc.This review describes the role of innate immunity in pathogenesis of psoriasis, and describes in detail the mechanisms involved into induction of inflammation of PAMPs and DAMPs. In psoriasis, positively charged catelicidin is considered one of the most important DAMPs, which can form a complex with negatively charged cell polyanions-LL-37/auto-RNA and LL-37/auto-DNA. The interaction of PAMP/DAMP ligands with specific PRR receptors leads to signal activation of effector components of immune system, i.e., assembly of inflammasome complex, caspase activation, synthesis of inflammatory cytokines and processing of their immature forms. The review focuses on the role of TLRs under the conditions of physiological norm, which recognize danger signals and provide protection from pathogens and their timely elimination, and in development of pathological process. Activation of TLRs induces the production of pro-inflammatory cytokines, interferons and antimicrobial peptides, chemokines that support the development of psoriatic inflammation.In addition to TLRs, the mechanisms of involvement of inflammasomes in the development of psoriasis, which provides processing of mature forms of IL-1β and IL-18, are described in detail. Mature forms of these cytokines mediate the development of inflammation in psoriatic focus. In addition, processing of these cytokines by caspases using the positive feedback mechanism provides an additional signal to activate transcriptional activity of their genes and contributes to perpetuated inflammation.The review presents data confirming participation of inflammasomes in the pathogenesis of psoriasis. Much attention is paid to description of pharmacological inhibitors of inflammasomes, which in the future may be the drugs of choice for treatment of inflammatory diseases. The study of molecular mechanisms of the innate immune system will reveal new approaches to prognosis and development of targeted therapy for psoriasis.

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Дудник, Олена. « «ПРОСВІТА» УМАНЩИНИ ЗА ЧАСІВ УКРАЇНСЬКОЇ ЦЕНТРАЛЬНОЇ РАДИ ». Уманська старовина, n^o 8 (30 décembre 2021) : 177–88. http://dx.doi.org/10.31499/2519-2035.8.2021.249967.

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Ключові слова: «Просвіта», Уманський повіт, Київська губернія, культурно-освітня політика, Центральна Рада. Анотація Події Української революції 1917-1921 рр. сприяли заснуванню просвітницьких організацій, які своїм основним завданням вбачали надання різної допомоги населенню на ниві культурно-освітніх справ. У статті досліджується процес утворення товариств «Просвіта» в Уманському повіті Київської губернії. З огляду на вагомість реалізованих проектів в статті увага присвячена добі Центральної Ради. Базуючись на архівних документах та матеріалах періодики, з’ясовано, що організаційні заходи з відродження просвітницького руху в Київській губернії були започатковані відразу після зміни політичного режиму. Головну увагу «Просвіти» краю приділяли праці в українських селах. У публікації встановлено, що просвітницькі організації в повіті почали виникати завдяки народній ініціативі, передусім проявам організаційних зусиль національно налаштованої місцевої інтелігенції та сільської молоді. Встановлено, що у більшості сіл Уманського повіту «Просвіти» виступали єдиними структурами, які проводили активну роботу серед населення, їх діяльність фокусувалася у культурницькій і освітній площині. Одним із головних завдань, що стояли перед просвітянами, було відкриття власних книгозбірень або бібліотек-читалень, придбання літератури і періодичних видань, організація курсів українознавства, поширення освітніх знань серед населення тобто все те, що могло сприяти пробудженню національної свідомості українців краю та їх об’єднанню. Фінансова допомога просвітянам краю надавалася органами місцевої влади, самоврядувань, окремими громадянами. Посилання 29 chervnia v m. Talnomu, 1917 – 29 chervnia v m. Talnomu…1917 [June 29 in Talne …1917]. «Prosvita». Nova Rada. (Kyiv). 2 lypnia. [in Ukrainian]. Adamskyi, 2018 – Adamskyi V. R. «Prosvity» Podillia v dobu Ukrainskoi Tsentralnoi Rady (berezen 1917 – kviten 1918 rr.) [«Enlightenment» Podillya in the days of the Ukrainian Central Council (March 1917 - April 1918)] : Doslidzhennia. Dokumenty. Materialy. Khmelnytskyi: FOP Tsiupak A. A., 2018. 478 s. [in Ukrainian]. V Shukaivodi, 1917 – V Shukaivodi… 1917 [In Shukaivoda … 1917] – «Prosvita». Nova Rada. (Kyiv).28 kvitnia. [in Ukrainian]. Verstyuk, ta in., 2004 – Verstiuk V., Horobets V., Tolochko O. Ukraina i Rosiia v istorychnii retrospektyvi. Ukrainski proekty v Rosiiskii imperii [Ukraine and Russia in historical retrospective review. The Ukrainian projects in the Russian empire]. K., 2004. 504 s. [in Ukrainian]. Vynnychenko, 2007 – Vynnychenko V. Vidrodzhennia natsii. Reprynt. vidtvor. vyd. 1920 r. [Revival of the nation]: u 3 ch. K. : Vyd-vo polit. l-ry Ukrainy, 1990. Ch. III. 542 s. [in Ukrainian]. Herman, 1995 – Herman O. M. Diialnist tovarystva «Prosvita» na Podilli naprykintsi XIX I v pershii polovyni XX stolittia [Activities of the society «Enlightenment» in Podolia in the late XIX and early XX century]: dys. ... kand. ist. nauk: 07.00.01. Chernivtsi, 1995. 228 s. [in Ukrainian]. Hrytsak, 1996 – Hrytsak Ya. Narys istorii Ukrainy. Formuvannia modernoi ukrainskoi natsii u XIX-XX st. [Essays on the history of Ukraine: the formation of the modern Ukrainian nation of the XIX-XX centuries]. K.: Heneza, 1996. 358 s. [in Ukrainian]. DAKO – Derzhavnyi arkhiv Kyivskoi oblasti Doroshenko, 2007 – Doroshenko D. Moi spomyny pro nedavnie mynule (1914-1920 roky) [My memories of the recent past (1914-1920)]. K. : Tempora, 2007. 632 s. [in Ukrainian]. Kravchuk, 1996 – Kravchuk L. V. Kulturo-tvorcha diialnist ta prosvitnytskyi rukh v period Ukrainskoi derzhavnosti 1917–1920 rr. [Cultural activity and educational movement in the period of Ukrainian statehood 1917-1920]: avtoref. dys. ... kand. ist. nauk: 07.00.01. Chernivtsi, 1996. 23 s. [in Ukrainian]. Kulturno-prosvitnii hurtok, 1918 – Kulturno-prosvitnii hurtok…1918 [Cultural and educational circle…1918]. «Prosvita». Nova Rada. (Kyiv). 25 kvitnia. [in Ukrainian]. Lozovyi, 2006 – Lozovyi V. S. Poshyrennia prosvitnytskykh oseredkiv v ukrainskomu seli v period Tsentralnoi Rady (1917 r.) [Dissemination of educational centers in the Ukrainian countryside during the Central Rada (1917)]. Osvita, nauka i kultura na Podilli: zb. nauk. pr. / hol. red. kol.: P. T. Tronko. Kam’ianets-Podilskyi: Oiium, 2006. T. 7: mat. tretoho kruhloho stolu «Kultura, osvita i prosvitnytskyi rukh na Podilli u KhVIII – na pochatku KhKhI st.». S. 3-11. [in Ukrainian]. Ostashko,1997 – Ostashko T. Tovarystvo «Prosvita» – oseredky ukrainskoho natsionalno-osvitnoho rukhu za doby Tsentralnoi Rady [Society «Enlightenment» is a center of the Ukrainian national educational movement during the Central Rada] // Tsentralna Rada i ukrainskyi derzhavotvorchyi protses (do 80-richchia stvorennia Tsentralnoi Rady): Materialy nauk. konf., 20 bereznia 1997 r. NAN Ukrainy, In-t istorii Ukrainy. K., 1997. Ch. 2. S. 272-280. [in Ukrainian]. Reient, 2003 – Reient O. Ukraina v impersku dobu (XIX – pochatok XX st.) [Ukraine in the imperial era (XIX - early XX centuries)]. K., 2003. 338 s. [in Ukrainian]. Sarbei, 1999 – Sarbei V. Natsionalne vidrodzhennia Ukrainy [National revival of Ukraine]. K., 1999. 335 s. [in Ukrainian]. U mistechku Pokotylove, 1917 – U mistechku Pokotylove…1917 [In the town of Pokotylove…1917] – Chytalnia. Nova Rada. (Kyiv). 26 chervnia. [in Ukrainian]. U s. Tykhomu Khutori, 1917 – U s. Tykhomu Khutori…1917 [In the village of Tykhy Khutir] – «Prosvita». Nova Rada. (Kyiv). 1 zhovtnia. [in Ukrainian]. Faryna, 1993 – Faryna S. Ya. Rol «Prosvit» v ukrainskomu natsionalno-kulturnomu rusi na pochatku KhKh stolittia [The role of «Enlightenment» in the Ukrainian national and cultural movement in the early XIX century].: dys. ... kand. ist. nauk: 07.00.01. Kremenchuk, 1993. 233 s. [in Ukrainian]. TsDAVO Ukrainy – Tsentralnyi derzhavnyi arkhiv vyshchykh orhaniv vlady ta upravlinnia Ukrainy.

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Ghassemi, Mehdi. « Possibility of Self-Representation : the Assimilation of Paul de Man’s Theories in John Banville’s Shroud ». Mosaïque, n^o 8 (1 juin 2013). http://dx.doi.org/10.54563/mosaique.1529.

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Shroud de John Banville, publié en 2002, peut être lu comme un exemple de « métafiction historiographique ». Le roman fonde en effet son protagoniste, Axel Vander, sur la vie de Paul de Man, le théoricien littéraire, dont les écrits dans un journal pro-nazi ont été découvert à titre posthume par Ortwin de Graef. Ce récit prend la forme d'une confession autobiographique, tout en s'appuyant sur les théories de Paul de Man concernant la question de la référentialité et le problème d '(auto-) représentation. En outre, selon de Man, le discours de l'autobiographie implique un processus d’« auto-restauration » par lequel le sujet confère à lui-même un masque dans le but de cacher l'absence d’un moi authentique et cohérent. Cependant, un problème se pose lorsque la langue devient le support de cette représentation (DE MAN, 1979) : loin de refléter le « vrai » moi, la langue devient le moyen même de la fabrication de masques. En résulte un « malaise» d'indécidabilité pour le narrateur. L’objectif de cet article est d'analyser Shroud en le lisant avec le texte de de Man intitulé « L'autobiographie en tant que de-facement » et de montrer comment le langage vient se poser comme l'agent qui déstabilise le discours même qu’il vise à restaurer. Après avoir démontré comment l'autobiographie entraîne une crise de l’autorité et de la représentation pour le narrateur, nous tâcherons, cependant, de démontrer qu’Axel peut aparaître comme un « noyau irréductible », ce qui subsiste à ce que de Man appelle « a linguistic predicament » Je constaterai également que ce noyau est incarné par la voix en tant que dimension du réel lacanien.

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Sharma, Deepak Kumar, Bincy Joseph, Rajesh Singathia, Abhishek Gaurav et Vishnu Kumar. « Inflammasome Activation and Pro-inflammatory Cytokine Genes Expression Following Exposure of PPRV in Vero Cells ». Indian Journal of Animal Research, Of (14 juillet 2023). http://dx.doi.org/10.18805/ijar.b-5102.

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Background: Peste des petits ruminants (PPR), a highly contagious and fatal disease of small ruminants, caused by PPR virus (PPRV). Detection of microbes is mediated by pattern recognition receptors (PRRs). These PRRs induce the assembly of a multi-protein signaling platform called the Inflammasome. It detects the pathogens and induces the synthesis of pro-inflammatory cytokines interleukins (IL) i.e. IL-1β and IL-18. The aim of this study was to assess the activation of inflammasome related genes, following exposure to PPRV in Vero Cells and was to assess the effect of activation of inflammasome on downstream pro-inflammatory cytokines IL-1β and IL-18 gene expression. Methods: Vero cells were infected with PPRV. The cells were incubated for different periods. Total RNA was reverse transcribed and used for amplification by quantitative real time PCR (qRT-PCR) of NLRP3, ASC, Caspase-1 and cytokines IL-1β and IL-18 gene expression. Result: It was revealed that 1 MOI of PPRV was appropriate for NLRP3 inflammasome activation. NLRP3 and ASC showed an increased level of expression at 2-4 hrs post inoculation (hpi). While, the Caspase-1 showed biphasic expression on 2-4 and 24 hpi, further, in the synchrony to it, IL-1β and IL-18 expression were also found to increase at 2-4 and 24 hpi.

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Cheng, You-Hung, Chun-Te Wu, Chih-Hao Chiu, Kuo-Yao Hsu, Shih-Sheng Chang, Yi-Sheng Chan et Alvin Chao-Yu Chen. « A Comparative Study on Arthroscopic Superior Capsular Reconstruction Using Fascia Lata Autograft With and Without Long Head of the Biceps Tendon Augmentation : Two-Year Patient-Reported Outcomes and Radiographic Analysis ». Orthopaedic Journal of Sports Medicine 12, n^o 10 (octobre 2024). http://dx.doi.org/10.1177/23259671241270243.

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Background: Given the growing concerns regarding objective measures of clinical outcomes, attention has recently been devoted to the establishment of clinically significant outcome (CSO) thresholds for patient-reported functional scores after rotator cuff surgery. Purpose: To retrospectively compare patient-reported outcome (PRO) measures (PROMs) and radiographic data between patients who underwent arthroscopic superior capsular reconstruction (SCR) with and without long head of the biceps tendon (LHBT) augmentation. Study Design: Cohort study; Level of evidence, 3. Methods: A total of 43 patients receiving arthroscopic SCR between 2016 and 2020 were enrolled, including a biceps augmentation group (n = 27) and a nonaugmentation group (n = 16). Patients were asked an anchor question regarding their satisfaction and perception of improvements. PROMs of American Shoulder and Elbow Surgeons (ASES), Constant score, Single Assessment Numeric Evaluation (SANE), and visual analog scale (VAS) for pain scores and radiographic data including magnetic resonance imaging and plain radiographs were collected and compared between the 2 groups. Anchor questions in CSO analysis for deriving the minimal clinically importance difference (MCID), substantial clinical benefit (SCB), Patient Acceptable Symptom State (PASS), and maximal outcome improvement (MOI) values were applied ≥2 years postoperatively. Results: Based on satisfaction responses, 17 patients were classified as satisfied, 16 as unsatisfied, and 10 as fair. Additionally, 13 patients felt they were improved, 14 changed, and 16 unchanged. Intergroup comparison based on patients’ satisfaction and perception of change or improvement exhibited significant differences in all 4 functional scores in favor of the satisfied and improved patients. However, there was no significant difference in the ΔVAS scores between the groups. CSO analyses showed no significant difference in percentage of patients achieving MCID, SCB, and PASS thresholds for the ΔASES, ΔConstant, and ΔSANE scores between patients undergoing arthroscopic SCR with or without LHBT augmentation. A significant difference was found in the percentage of patients achieving the MOI for ΔASES score with 70.4% in the augmented group and 37.5% in the nonaugmented group, respectively. The mean acromiohumeral distance (AHD) differed significantly between augmentation (8.1 ± 2.2 mm) and nonaugmentation (7 ± 1.9 mm) groups. The graft tear rate did not differ significantly. Conclusion: There was no significant difference in PROs and percentage of patients achieving MCID, SCB, and PASS between isolated and augmented SCR groups. A higher percentage of patients achieving MOI and slightly greater AHD were found in the augmented group. Further evaluation is required to determine if there is any long-term benefit to LHBT augmentation of SCR.

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Marín-Palma, Damariz, Jorge H. Tabares-Guevara, María I. Zapata-Cardona, Wildeman Zapata-Builes, Natalia Taborda, Maria T. Rugeles et Juan C. Hernandez. « PM10 promotes an inflammatory cytokine response that may impact SARS-CoV-2 replication in vitro ». Frontiers in Immunology 14 (25 avril 2023). http://dx.doi.org/10.3389/fimmu.2023.1161135.

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IntroductionIn the last decades, a decrease in air quality has been observed, mainly associated with anthropogenic activities. Air pollutants, including particulate matter (PM), have been associated with adverse effects on human health, such as exacerbation of respiratory diseases and infections. High levels of PM in the air have recently been associated with increased morbidity and mortality of COVID-19 in some regions of the world.ObjectiveTo evaluate the effect of coarse particulate matter (PM10) on the inflammatory response and viral replication triggered by SARS-CoV-2 using in vitro models.MethodsPeripheral blood mononuclear cells (PBMC) from healthy donors were treated with PM10 and subsequently exposed to SARS-CoV-2 (D614G strain, MOI 0.1). The production of pro-inflammatory cytokines and antiviral factors was quantified by qPCR and ELISA. In addition, using the A549 cell line, previously exposed to PM, the viral replication was evaluated by qPCR and plaque assay.ResultsSARS-CoV-2 stimulation increased the production of pro-inflammatory cytokines in PBMC, such as IL-1β, IL-6 and IL-8, but not antiviral factors. Likewise, PM10 induced significant production of IL-6 in PBMCs stimulated with SARS-CoV-2 and decreased the expression of OAS and PKR. Additionally, PM10 induces the release of IL-1β in PBMC exposed to SARS-CoV-2 as well as in a co-culture of epithelial cells and PBMCs. Finally, increased viral replication of SARS-CoV-2 was shown in response to PM10.ConclusionExposure to coarse particulate matter increases the production of pro-inflammatory cytokines, such as IL-1β and IL-6, and may alter the expression of antiviral factors, which are relevant for the immune response to SARS-CoV-2. These results suggest that pre-exposure to air particulate matter could have a modest role in the higher production of cytokines and viral replication during COVID-19, which eventually could contribute to severe clinical outcomes.

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Somayajulu, Mallika, Sharon A. McClellan, Farooq Muhammed, Robert Wright et Linda D. Hazlett. « PM10 and Pseudomonas aeruginosa : effects on corneal epithelium ». Frontiers in Cellular and Infection Microbiology 13 (5 octobre 2023). http://dx.doi.org/10.3389/fcimb.2023.1240903.

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PurposeIn vivo data indicate that mouse corneas exposed to PM10 showed early perforation and thinning after infection with Pseudomonas aeruginosa. To understand the mechanisms underlying this finding, we tested the effects of PM10 and the mitochondria targeted anti-oxidant SKQ1 in immortalized human corneal epithelial cells (HCET) that were challenged with Pseudomonas aeruginosa strain 19660.MethodsMouse corneas were infected with strain 19660 after a 2 week whole-body exposure to PM10 or control air and assessed by clinical scores, slit lamp photography and western blot. HCET were exposed to 100μg/ml PM10 for 24h before challenge with strain 19660 (MOI 20). A subset of cells were pre-treated with 50nM SKQ1 for 1h before PM10 exposure. Phase contrast microscopy was used to study cell morphology, cell viability was measured by an MTT assay, and ROS by DCFH-DA. Levels of pro-inflammatory markers and anti-oxidant enzymes were evaluated by RT-PCR, western blot and ELISA. Reduced glutathione (GSH) and malondialdehyde (MDA) levels were evaluated by assay kits.ResultsIn vivo, whole body exposure to PM10 vs. control air exposed mouse corneas showed early perforation and/or corneal thinning at 3 days post infection, accompanied by increased TNF-α and decreased SOD2 protein levels. In vitro, PM10 induced a dose dependent reduction in cell viability of HCET and significantly increased mRNA levels of pro-inflammatory molecules compared to control. Exposure to PM10 before bacterial challenge further amplified the reduction in cell viability and GSH levels. Furthermore, PM10 exposure also exacerbated the increase in MDA and ROS levels and phase contrast microscopy revealed more rounded cells after strain 19660 challenge. PM10 exposure also further increased the mRNA and protein levels of pro-inflammatory molecules, while anti-inflammatory IL-10 was decreased. SKQ1 reversed the rounded cell morphology observed by phase contrast microscopy, increased levels of MDA, ROS and pro-inflammatory molecules, and restored IL-10.ConclusionsPM10 induces decreased cell viability, oxidative stress and inflammation in HCET and has an additive effect upon bacterial challenge. SKQ1 protects against oxidative stress and inflammation induced by PM10 after bacterial challenge by reversing these effects. The findings provide insight into mechanisms underlying early perforation and thinning observed in infected corneas of PM10 exposed mice.

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Zhang, Wei, Zhixing Li, Haitao Yang, Guanglu Wang, Gang Liu, Yu Wang, Babatunde Kazeem Bello, Panpan Zhao, Wei Liang et Jingquan Dong. « Aeromonas sobria Induces Proinflammatory Cytokines Production in Mouse Macrophages via Activating NLRP3 Inflammasome Signaling Pathways ». Frontiers in Cellular and Infection Microbiology 11 (26 août 2021). http://dx.doi.org/10.3389/fcimb.2021.691445.

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Aeromonas sobria, a common conditional pathogenic bacteria, is widely distributed in the environment and causes gastroenteritis in humans or septicemia in fish. Of all Aeromonas species, A. sobria is the most frequently isolated from human infections especially in immunocompromised subjects. Innate immunity is the first protection system of organism to resist non-specific pathogens invasion; however, the immune response process of hosts against A. sobria infection re\mains unexplored. The present study established an A. sobria infection model using primary mouse peritoneal macrophages (PMφs). The adherence and cytotoxicity of A. sobria on PMφs were determined by May-Grünwald Giemsa staining and LDH release measurement. Pro-inflammatory cytokine expression levels were measured using qPCR, western blotting, and ELISA methods. We also investigated the levels of ASC oligomerization and determined the roles of active caspase-1 in IL-1β secretion through inhibition assays and explored the activated pattern recognition receptors through immunofluorescence. We further elucidated the roles of activated inflammasome in regulating the host’s inflammatory response through inhibition combined with ELISA assays. Our results showed that A. sobria induced lytic cell death and LDH release, whereas it had no adhesive properties on PMφs. A. sobria triggered various proinflammatory cytokine transcription level upregulation, and IL-1β occupied the highest levels. The pro-IL-1β protein expression levels increased in a dose-dependent manner with MOI ranging from 1 to 100. This process was regulated by ASC-dependent inflammasome, which cleavage pro-IL-1β into active IL-1β p17 with activated caspase-1 p20. Meanwhile, the expression levels of NLRP3 receptor significantly increased, location analysis revealed puncta-like surrounding nuclear, and inhibition of NLRP3 inflammasome downregulated caspase-1 activation and IL-1β secretion. Blocking of NLRP3 inflammasome activation through K+ efflux and cathepsin B or caspase approaches downregulated A. sobria–induced proinflammatory cytokine production. Overall, these data indicated that A. sobria induced proinflammatory cytokine production in PMφs through activating NLRP3 inflammasome signaling pathways.

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Pitaloka, Dian, et Muti’ah Nurul Jihadah. « Comparing Responses of Ursolic Acid in Murine Macrophages Infected with Mycobacterium smegmatis and Mycobacterium avium ». Indonesian Journal of Pharmacy, 20 décembre 2022, 602–9. http://dx.doi.org/10.22146/ijp.3507.

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Mycobacterium smegmatis and Mycobacterium avium offer an advantage in examining tuberculosis-like effects and host immune defense. Therefore, the study aims to examine the effect of ursolic acid (UA) on the host immune system by analyzing cytokines concentration, such as TNF-α, IL-6, IL-1β, and nitrite oxide produced by murine macrophages infected with Mycobacterium smegmatis and Mycobacterium avium. Femurs of female C57BL/6 mice aged 6–8 weeks were used to culture the Bone marrow-derived macrophages (BMDM). On day 10, BMDM was infected with Mycobacterium smegmatis and Mycobacterium avium using a multiplicity of infection (MOI) amounting to 8:1, then TNF-α, IL-6, and IL-1β were analyzed using ELISA and nitrite oxide with Griess reagent. The results showed that UA decreased the production of three respective pro-inflammatory cytokines used in the study, both in BMDM infected by Mycobacterium smegmatis and Mycobacterium avium. For TNF-α, the reduction was nearly 65%-90% compared to the control. The decrease in the production of IL-6 occurred from 2700 pg/ml to 750 pg/ml for BMDM infected with Mycobacterium smegmatis, while the reduction was more significant in those infected using Mycobacterium avium with approximately 150 pg/ml compared to the control. Moreover, UA reduced by over 90% of IL-1β and this result was in line with the reduction of nitrite. UA decreases the production of pro-inflammatory cytokines such as TNF-α, IL-6, IL-1β, and nitrite. This result is preliminary but supports further study on the role of UA in immune defense from pathogenic and non-pathogenic mycobacterial infections.

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Evani, Shankar J., Shatha Dallo et Anand K. Ramasubramanian. « Abstract 16534 : Remodeling of Intimal Matrix by C. Pneumoniae-infected Macrophages ». Circulation 132, suppl_3 (10 novembre 2015). http://dx.doi.org/10.1161/circ.132.suppl_3.16534.

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Introduction: Chlamydia pneumoniae, an intracellular pulmonary pathogen, is considered an important risk factor for atherosclerosis through the mechanisms is poorly understood. Infected monocytes from the lung enter atherosclerotic foci, and create a pro-inflammatory environment in the intima. Here, we tested the hypothesis that C. pneumoniae infection contributes to intimal matrix remodeling using a 3D model. Methods: Primary human monocytes infected with C. pneumoniae (either live of heat-killed organisms) at an MOI-1 were embedded in type I collagen matrix. Between 1 to 9 days, we analyzed the kinetics of MMP and cytokine release, cell migration, and structural and mechanical characteristics of the matrix by ELISA, microscopy and rheometry, respectively – with or without specific inhibitors of molecular mechanisms. All experiments were performed with at least 3 donors in triplicates; and statistical significance was assessed by Student’s t -test or ANOVA. Results: We observed that the infection of macrophages encapsulated in a 3D collagen matrix promote: (i) the production of MMP and pro-inflammatory cytokines; (ii) the remodeling process including degradation, aggregation and realignment of fibers (Fig. A); (iii) an increase in macrophage podosomal extensions and microenvironmental interactions; (iv) the collective, coordinated migration of macrophages to form focal clusters (Fig. B); and (v) an increase in the elastic modulus (Fig. C). We observed that these changes were more pronounced in live than in heat-killed infection. Further, inhibition of MMP, chemotactic stimuli and cell migratory signaling attenuates matrix remodeling. Significance: Our results provide novel insights into the role of infection and the mechanisms in modulating atherosclerotic progression, which may provide an attractive new approach to not only test the infectious burden hypothesis but also for the prevention and/or treatment of cardiovascular diseases.

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Evani, Shankar J., Shatha F. Dallo et Anand K. Ramasubramanian. « Abstract 473 : Fluid Shear Stress Specifically Exacerbates Chlamydia pneumoniae-mediated Pro-inflammatory Responses in Monocytes ». Arteriosclerosis, Thrombosis, and Vascular Biology 33, suppl_1 (mai 2013). http://dx.doi.org/10.1161/atvb.33.suppl_1.a473.

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Background and Significance The precise role of Chlamydia pneumoniae infection in various stages of the atherosclerosis is not well understood. During the transit from lung to the atherosclerotic sites, C. pneumoniae-infected monocytes travel through circulation and are subjected to shear stress due to blood flow. Elucidating the effects of these mechanical stimuli on infected monocytes is critical in our understanding of the link between C. pneumoniae infection and atherosclerosis. Objectives We hypothesized that fluid shear stress alters the inflammatory response of C. pneumoniae-infected monocytes. Using an in vitro model of blood flow, we determined the effect of physiological levels of shear stress on monocyte responses pertinent to atherosclerosis including cytokine secretion, adhesion molecule expression, intracellular signaling, and endothelial adhesion. Methods Primary human monocytes and THP-1 cells were infected with C. pneumoniae at an MOI-2 for 2 h at 35 °C with intermittent rocking, and cultured for 36 h. The infected cells were then subjected to a shear stress of 7.5 dyn/cm2 for 1 h, and the supernatants were analyzed for a panel of 17 cytokines, and the cells were analyzed for signaling proteins, surface receptors, and endothelial adhesion. Uninfected cells and static conditions were used as controls, and a P<0.05 (Student’s t-test) was considered significant. Results We observed that shear stress on infected cells resulted in a caspase-independent upregulation of pro-inflammatory IL-1β (3-fold), a modest increase in MIP-1α and MIP-1β, a decrease in anti-inflammatory IL-10, and no change in other cytokines including TNFα and IL-12. These changes were manifested as an increase in the adhesion of monocytes to endothelium activated with the infected supernatants. We also observed differences in the chemotaxis of monocytes exposed to sheared vs. static supernatants. Conclusions We observe that physiological shear stress drives Chlamydia infected monocytes towards a pro-inflammatory state in circulation suggesting a critical synergy between mechanical and chemical factors in atherogenesis.

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Bishku, Michael B. « The Relations of Vietnam with the Middle East-North Africa Region : From a Divided State to an Important Member of the Association of Southeast Asian Nations ». Contemporary Review of the Middle East, 5 juillet 2023. http://dx.doi.org/10.1177/23477989231179259.

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During the Cold War, three countries were partitioned into two states: Germany, Korea, and Vietnam. The last was a result of its war of independence against the French following World War II and continued until 1954. Following a victorious war against the United States-backed government in the south, the communist government in the north reunited the country between 1975 and 1976. In 1988, facing economic troubles, Vietnam instituted Doi Moi market reforms while relaxing its ideological worldview and expanded its diplomatic and economic relations. In 1995, Vietnam joined the Association of Southeast Asian Nations (ASEAN), which it had regarded as a pro-Western organization during the Cold War. Meanwhile, first North Vietnam and later a reunified Vietnam went from a country whose relations were exclusive with states in the communist bloc and a few other non-aligned countries to one that today has ties with almost every country in the world. Academic studies on Vietnam’s foreign relations largely focus on those with Russia, China, the US, its ASEAN partners, and other countries in Asia. However, since the end of the Cold War, countries of the Middle East and North Africa (MENA) region are an import source of Vietnam’s crude oil and natural gas and an expanding market for its exports, while MENA states, in this era of globalization, have sought to increase diplomatic and economic ties throughout Asia.

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Bueno, Manuela R., Karin H. Ishikawa, Gislane Almeida-Santos, Ellen S. Ando-Suguimoto, Natali Shimabukuro, Dione Kawamoto et Marcia P. A. Mayer. « Lactobacilli Attenuate the Effect of Aggregatibacter actinomycetemcomitans Infection in Gingival Epithelial Cells ». Frontiers in Microbiology 13 (4 mai 2022). http://dx.doi.org/10.3389/fmicb.2022.846192.

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Probiotics may be considered as an additional strategy to achieve a balanced microbiome in periodontitis. However, the mechanisms underlying the use of probiotics in the prevention or control of periodontitis are still not fully elucidated. This in vitro study aimed to evaluate the effect of two commercially available strains of lactobacilli on gingival epithelial cells (GECs) challenged by Aggregatibacter actinomycetemcomitans. OBA-9 GECs were infected with A. actinomycetemcomitans strain JP2 at an MOI of 1:100 and/or co-infected with Lactobacillus acidophilus La5 (La5) or Lacticaseibacillus rhamnosus Lr32 (Lr32) at an MOI of 1:10 for 2 and 24 h. The number of adherent/internalized bacteria to GECs was determined by qPCR. Production of inflammatory mediators (CXCL-8, IL-1β, GM-CSF, and IL-10) by GECs was determined by ELISA, and the expression of genes encoding cell receptors and involved in apoptosis was determined by RT-qPCR. Apoptosis was also analyzed by Annexin V staining. There was a slight loss in OBA-9 cell viability after infection with A. actinomycetemcomitans or the tested probiotics after 2 h, which was magnified after 24-h co-infection. Adherence of A. actinomycetemcomitans to GECs was 1.8 × 107 (± 1.2 × 106) cells/well in the mono-infection but reduced to 1.2 × 107 (± 1.5 × 106) in the co-infection with Lr32 and to 6 × 106 (± 1 × 106) in the co-infection with La5 (p < 0.05). GECs mono-infected with A. actinomycetemcomitans produced CXCL-8, GM-CSF, and IL-1β, and the co-infection with both probiotic strains altered this profile. While the co-infection of A. actinomycetemcomitans with La5 resulted in reduced levels of all mediators, the co-infection with Lr32 promoted reduced levels of CXCL-8 and GM-CSF but increased the production of IL-1β. The probiotics upregulated the expression of TLR2 and downregulated TLR4 in cells co-infected with A. actinomycetemcomitans. A. actinomycetemcomitans-induced the upregulation of NRLP3 was attenuated by La5 but increased by Lr32. Furthermore, the transcription of the anti-apoptotic gene BCL-2 was upregulated, whereas the pro-apoptotic BAX was downregulated in cells co-infected with A. actinomycetemcomitans and the probiotics. Infection with A. actinomycetemcomitans induced apoptosis in GECs, whereas the co-infection with lactobacilli attenuated the apoptotic phenotype. Both tested lactobacilli may interfere in A. actinomycetemcomitans colonization of the oral cavity by reducing its ability to interact with gingival epithelial cells and modulating cells response. However, L. acidophilus La5 properties suggest that this strain has a higher potential to control A. actinomycetemcomitans-associated periodontitis than L. rhamnosus Lr32.

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Wherry, Taylor L. T., Rohana P. Dassanayake, John P. Bannantine, Shankumar Mooyottu et Judith R. Stabel. « Vitamin D3 alters macrophage phenotype and endosomal trafficking markers in dairy cattle naturally infected with Mycobacterium avium subsp. paratuberculosis ». Frontiers in Cellular and Infection Microbiology 12 (5 octobre 2022). http://dx.doi.org/10.3389/fcimb.2022.1021657.

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Macrophages are important host defense cells in ruminant paratuberculosis (Johne’s Disease; JD), a chronic enteritis caused by Mycobacterium avium subsp. paratuberculosis (MAP). Classical macrophage functions of pathogen trafficking, degradation, and antigen presentation are interrupted in mycobacterial infection. Immunologic stimulation by 25-hydroxyvitamin D3 (25(OH)D3) and 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) enhances bovine macrophage function. The present study aimed to investigate the role of vitamin D3 on macrophage phenotype and endosomal trafficking of MAP in monocyte-derived macrophages (MDMs) cultured from JD-, JD+ subclinical, and JD+ clinically infected cattle. MDMs were pre-treated 100 ng/ml 25(OH)D3 or 4 ng/ml 1,25(OH)2D3 and incubated 24 hrs with MAP at 10:1 multiplicity of infection (MOI). In vitro MAP infection upregulated pro-inflammatory (M1) CD80 and downregulated resolution/repair (M2) CD163. Vitamin D3 generally decreased CD80 and increased CD163 expression. Furthermore, early endosomal marker Rab5 was upregulated 140× across all stages of paratuberculosis infection following in vitro MAP infection; however, Rab5 was reduced in MAP-activated MDMs from JD+ subclinical and JD+ clinical cows compared to healthy controls. Rab7 expression decreased in control and clinical cows following MDM infection with MAP. Both forms of vitamin D3 reduced Rab5 expression in infected MDMs from JD- control cows, while 1,25(OH)2D3 decreased Rab7 expression in JD- and JD+ subclinical animals regardless of MAP infection in vitro. Vitamin D3 promoted phagocytosis in MDMs from JD- and JD+ clinical cows treated with either vitamin D3 analog. Results from this study show exogenous vitamin D3 influences macrophage M1/M2 polarization and Rab GTPase expression within MDM culture.

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Kumar, Parveen, Kanchan Saini, Vikram Saini et Tanecia Mitchell. « Oxalate Alters Cellular Bioenergetics, Redox Homeostasis, Antibacterial Response, and Immune Response in Macrophages ». Frontiers in Immunology 12 (21 octobre 2021). http://dx.doi.org/10.3389/fimmu.2021.694865.

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Individuals with calcium oxalate (CaOx) kidney stones can have secondarily infected calculi which may play a role in the development of recurrent urinary tract infection (UTI). Uropathogenic Escherichia coli (UPEC) is the most common causative pathogen of UTIs. Macrophages play a critical role in host immune defense against bacterial infections. Our previous study demonstrated that oxalate, an important component of the most common type of kidney stone, impairs monocyte cellular bioenergetics and redox homeostasis. The objective of this study was to investigate whether oxalate compromises macrophage metabolism, redox status, anti-bacterial response, and immune response. Monocytes (THP-1, a human monocytic cell line) were exposed to sodium oxalate (soluble oxalate; 50 µM) for 48 hours prior to being differentiated into macrophages. Macrophages were subsequently exposed to calcium oxalate crystals (50 µM) for 48 hours followed by UPEC (MOI 1:2 or 1:5) for 2 hours. Peritoneal macrophages and bone marrow-derived macrophages (BMDM) from C57BL/6 mice were also exposed to oxalate. THP-1 macrophages treated with oxalate had decreased cellular bioenergetics, mitochondrial complex I and IV activity, and ATP levels compared to control cells. In addition, these cells had a significant increase in mitochondrial and total reactive oxygen species levels, mitochondrial gene expression, and pro-inflammatory cytokine (i.e. Interleukin-1β, IL-1β and Interleukin-6, IL-6) mRNA levels and secretion. In contrast, oxalate significantly decreased the mRNA levels and secretion of the anti-inflammatory cytokine, Interleukin-10 (IL-10). Further, oxalate increased the bacterial burden of primary macrophages. Our findings demonstrate that oxalate compromises macrophage metabolism, redox homeostasis, and cytokine signaling leading to a reduction in anti-bacterial response and increased infection. These data highlight a novel role of oxalate on macrophage function.

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Li, Yongjian, Qiong Chen, Shasha Wang et Shichao Zhang. « Ligustrazine Attenuates Gastric Inflammation and Apoptosis in Helicobacter pylori-induced Gastric Epithelial Cells ». Jundishapur Journal of Microbiology 14, n^o 4 (27 juillet 2021). http://dx.doi.org/10.5812/jjm.116612.

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Background: Stomach disorders, including gastric cancer and gastritis, are associated with the pathogenic bacterium Helicobacter pylori. Enhanced inflammation is the characteristic of H. pylori-induced gastritis. Ligustrazine exerts anti-inflammatory properties in mouse asthma models and acute kidney injury. Objectives: To determine the role of ligustrazine in H. pylori-induced gastritis. Methods: Normal gastric epithelial cell line (GES-1) was cultured with H. pylori at a multiplicity of infection (MOI) of 100: 1 for 24 hours. GES-1 cell line under H. pylori condition was incubated with 100 or 200 μM ligustrazine for 24 hours. Cell viability and apoptosis were investigated by MTT and flow cytometry assays, respectively. Inflammation was assessed by determining the levels and mRNA expression of interleukins (IL)-6/8, tumor necrosis factor-α (TNF-α), and monocyte chemotactic protein 1 (MCP-1) using ELISA and qRT-PCR analysis, respectively. Results: Helicobacter pylori infection reduced the viability and promoted the apoptosis of GES-1 cell line, accompanied by the enhanced activities of caspases 3 and 9. However, ligustrazine reversed the H. pylori-induced infection decreased viability, while increased apoptosis and caspases 3/9 activities in GES-1 cell line. Moreover, ligustrazine attenuated H. pylori-induced secretions of pro-inflammatory factors, IL-6/8, TNF-α, and MCP-1, in GES-1 cell line. The protein expression of inhibitor of NF-κB (IκBα) was downregulated in GES-1 cell line after H. pylori infection, while the protein expression levels of p65 and phosphorylation of IκBα were upregulated by H. pylori infection. On the contrary, ligustrazine decreased H. pylori-induced protein expression of IκBα, whereas increased protein expression of p65 and phosphorylation of IκBα. Conclusions: Ligustrazine exerted protective effects on H. pylori-induced gastric epithelial cells through inhibition of gastric inflammation and apoptosis and inactivation of NF-κB pathway.

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Pandey, Neha, Meghana Rastogi et Sunit K. Singh. « Chandipura virus dysregulates the expression of hsa-miR-21-5p to activate NF-κB in human microglial cells ». Journal of Biomedical Science 28, n^o 1 (7 juillet 2021). http://dx.doi.org/10.1186/s12929-021-00748-0.

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Abstract Background Chandipura virus (CHPV) is a negative single-stranded RNA virus of the Rhabdoviridae family. CHPV infection has been reported in Central and Western India. CHPV causes acute encephalitis with a case fatality rate of 70 % and mostly affects children below 15 years of age. CHPV infection in brain leads to neuronal apoptosis and activation of the microglial cells. The microRNAs (miRNAs) are small endogenous non-coding RNA that regulate the gene expression. Viral infections perturb the expression pattern of cellular miRNAs, which may in turn affect the expression pattern of downstream genes. This study aims to investigate hsa-miR-21-5p mediated regulation of PTEN, AKT, NF-ĸBp65, IL-6, TNF-α, and IL-1β, in human microglial cells during CHPV infection. Methods To understand the role of hsa-miR-21-5p in CHPV infection, the human microglial cells were infected with CHPV (MOI-0.1). Real-time PCR, western blotting, Luciferase assay, over-expression and knockdown techniques were used to understand the role of hsa-miR-21-5p in the regulation of PTEN, AKT and, NF-ĸBp65, IL-6, TNF-α, and IL-1β in this study. Results The hsa-miR-21-5p was found to be upregulated during CHPV infection in human microglial cells. This led to the downregulation of PTEN which promoted the phosphorylation of AKT and NF-ĸBp65. Over-expression of hsa-miR-21-5p led to the decreased expression of PTEN and promoted further phosphorylation of AKT and NF-ĸBp65 in human microglial cells. However, the inhibition of hsa-miR-21-5p using hsa-miR-21-5p inhibitor restored the expression. Conclusions This study supports the role of hsa-miR-21-5p in the regulation of pro-inflammatory genes in CHPV infected human microglial cells.

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Wang, Jinxin, Qun Ding, Qiankun Yang, Hui Fan, Guili Yu, Feixue Liu, Babatunde Kazeem Bello et al. « Vibrio alginolyticus Triggers Inflammatory Response in Mouse Peritoneal Macrophages via Activation of NLRP3 Inflammasome ». Frontiers in Cellular and Infection Microbiology 11 (15 novembre 2021). http://dx.doi.org/10.3389/fcimb.2021.769777.

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Vibrio alginolyticus is a food-borne marine Vibrio that causes gastroenteritis, otitis media, otitis externa, and septicemia in humans. The pathogenic mechanisms of V. alginolyticus have previously been studied in aquaculture animals; however, the underlying mechanisms in mammals remain unknown. In this study, an in vitro model of mouse peritoneal macrophages infected with V. alginolyticus was established. qPCR results revealed that V. alginolyticus induced the transcription levels of various cytokines, including IL-1β, IL-12, IL-18, TNF-α, IL-17, IL-6, IFN-γ, and IL-10, and the secretion level of IL-1β is the most significant. Inhibition assays with Ac-YVAD-CHO (a caspase-1 inhibitor) and Z-VAD-FMK (a pan-caspase inhibitor) were conducted to determine whether caspase-1 or caspase-11 is involved in V. alginolyticus-triggered IL-1β secretion. Results showed that IL-1β secretion was partly inhibited by Ac-YVAD-CHO and absolutely blocked by Z-VAD-FMK. To explore the sensed pattern recognition receptors, several NLR family members and the AIM2 receptor were detected and many receptors were upregulated especially NLRP3. Moreover, the NLRP3 protein displayed a puncta-like surrounding cell nucleus, which signified that the NLRP3 inflammasome was activated in response to V. alginolyticus infection. Inhibition assays with glyburide and CA-074 methyl ester (K+ outflow inhibitor and cathepsin B inhibitor) blocked IL-1β secretion, which demonstrated the essential role of the NLRP3 inflammasome in inflammatory response. To better understand how V. alginolyticus affects IL-1β release, the NLRP3 inflammasome was detected with doses ranging from 0.1 to 10 MOIs and time periods ranging from 3 to 12 h. Results showed that V. alginolyticus-mediated NLRP3 inflammasome activation was in a time- and dose-dependent manner and IL-1β release peaked at MOI of 1 for 12 h. Most importantly, blocking the NLRP3 inflammasome with inhibitors and the use of NLRP3-/- and caspase-1/11-/- mice could attenuate pro-inflammatory cytokine secretion, such as IL-1β, IL-6, IL-12, and TNF-α. Taken together, our study first found that the NLRP3 inflammasome plays vital roles in V. alginolyticus triggered inflammatory response in mouse peritoneal macrophages. This may provide reference information for the development of potential anti-inflammatory treatments against V. alginolyticus infection.

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