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Kosinova, E., i I. Psikal. "Restriction fragment length polymorphism of ORF6 and ORF7 genes of porcine reproductive and respiratory syndrome virus (PRRSV) vaccine strains registered in the CzechRepublic". Veterinární Medicína 51, No. 8 (27.03.2012): 414–22. http://dx.doi.org/10.17221/5565-vetmed.
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Restriction fragment length polymorphism (RFLP) of open reading frames 6 and 7 was applied to comparative genetic analysis of live attenuated vaccine strains (Amervac-PRRS/A3, Porcilis PRRS, Ingelvac PRRS) of porcine reproductive and respiratory syndrome virus (PRRSV), registered in the Czech Republic, six field viruses (L-588, L-1606, L-2053, L-3305, L-6558, L-6791), and three PRRSV local field isolates (CAMP V-502, CAMP V-503, VOS 2878) found in pig herds in the Czech Republic and Slovak Republic. The set of restriction enzymes Hae II, Alu I and BsaJ I allowed the differentiation of local field isolates, field viruses of PRRS, and vaccine strains of the European genotype from North American genotype, but could also distinguish between viruses of the same genotype. Five different RFLP patterns were obtained from twelve examined PRRS viruses by combination of the above restriction enzymes. RFLP code 1-1-1 was the most frequent digestion pattern within all PRRS field viruses (L-588, L-1606, L-2053, L-3305, L-6558, L-6791), CAMP V-502 isolate and vaccine strain Porcilis PRRS, which is suggestive of higher antigenic identity among the compared viruses. In the North American types (Ingelvac PRRS vaccine strain and VOS 2878 isolate), homogeneity in restriction patterns (code 2-x-4) was recorded. These studies indicate that PCR-based RFLP analysis of ORF6 and ORF7 of genes might be a suitable tool in epidemiological studies of PRRSV, similarly to the studies based on genetic analysis of ORF5 gene.
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Bertram, Mark J., Steve J. Kitt i Keith Kinsely. "PSVI-14 Impact of Bacillus Subtilis and Bacillus Licheniformis on Weaning Weight and Subsequent Performance, Health and Immune Parameters When Challenged with PRRSV with and Without PRRS Vaccination". Journal of Animal Science 100, Supplement_2 (12.04.2022): 160–61. http://dx.doi.org/10.1093/jas/skac064.274.
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Abstract A study was conducted examining the impact of feeding BioPlus 2B to PRRSV naïve sows during gestation and lactation on litter performance and on post-weaning performance when challenged with PRRSV in vaccinated and unvaccinated pigs. A total of 108 sows were allotted by parity and received a control diet (C) or BioPlus 2B (B2B) supplemented diet beginning a minimum of 31 d prior to farrowing through weaning. Pig weight was measured at birth and 17 d of age. At weaning, a subset of 200 pigs from each treatment were transported to a nursery and randomly allotted within dietary treatment on day 1 to a control (-V) or Ingelvac PRRS MLV vaccine (+V), establishing treatments of C+V, C-V, B2B+V, and B2B-V. On d 21, all pigs were intramuscularly challenged with a 1-18-2 PRRS virus. Growth rate and feed intake were monitored, serological and immune measures were recorded on d 21, 28, 35 and 42, post-weaning and on d 42 gross lung lesions were recorded. Pigs from sows receiving B2B tended (P < 0.10) to weigh more at 17 d of age and 21 d post-weaning (Table 1). Prior to challenge, there were no performance differences among treatments. Post challenge, there was a V x B2B interaction with C+V, B2B+V and B2B-V pigs having higher ADG and FG (P < .05) compared with C-V. On day 42, pigs fed B2B displayed improved lung lesion scores (P < 0.05) regardless of vaccine status. PRRSV viremia, measured by PCR and qPCR decreased more quickly when pigs consumed B2B (P < 0.05). Additionally, B2B modulated PRRSV specific T-cell interferon gamma production (P < 0.05). In conclusion, pigs consuming B2B from sows fedB2B were heavier at weaning and when subjected to a PRRSV challenge post-weaning, grew faster, were more efficient, and had reduced gross lung lesions, likely due to changes in immune function.
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Indik, S., i L. Valíček. "Differentiation of porcine reproductive and respiratory syndrome virus European vaccine strains from Czech field isolates by restriction fragment length polymorphism analysis of ORF5 gene". Veterinární Medicína 47, No. 10 - 11 (30.03.2012): 295–301. http://dx.doi.org/10.17221/5838-vetmed.
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Restriction fragment length polymorphism (RFLP) analysis of open reading frame 5 was developed for typing of Czech strains of porcine reproductive and respiratory syndrome virus (PRRSV). The set of restriction enzymes Acc I, Hae II and SnaB I allowed the differentiation of heterogeneous Czech strains of PRRSV clustered separately in the phylogenetic tree. The high-passage strain V-502 (164) was also differentiated from its parent strain V-502. The same restriction enzymes could distinguish the European-type vaccine strains Porcilis PRRS and Pyrsvac-183, registered inCzechRepublic, from the Czech field isolates. The published ORF5 nucleotide sequences allowed us to presume that it will also be possible to distinguish most of European field strains from vaccine strains. PCR-based RFLP analysis can become a valuable tool in epidemiological studies of PRRSV inEurope.
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Torricelli, Martina, Anna Fratto, Marcella Ciullo, Carla Sebastiani, Chiara Arcangeli, Andrea Felici, Samira Giovannini, Francesca Maria Sarti, Marco Sensi i Massimo Biagetti. "Porcine Reproductive and Respiratory Syndrome (PRRS) and CD163 Resistance Polymorphic Markers: What Is the Scenario in Naturally Infected Pig Livestock in Central Italy?" Animals 13, nr 15 (31.07.2023): 2477. http://dx.doi.org/10.3390/ani13152477.
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Porcine Reproductive and Respiratory Syndrome (PRRS) caused by the PRRS virus affects farmed pigs worldwide, causing direct and indirect losses. The most severe manifestations of PRRS infection are observed in piglets and pregnant sows. The clinical outcome of the infection depends on the PRRSV strain’s virulence, the pregnancy state of the female, environmental factors, the presence of protective antibodies due to previous infections, and the host’s genetic susceptibility. The latter aspect was investigated in this study, in particular, evaluating the most significant polymorphisms (SNPs) of the CD163 gene in slaughtered pigs reared in Central Italy. Total RNAs were extracted from 377 swine samples and subjected to RT-PCR targeted to the CD163 gene, followed by sequencing analysis. Contextually, the viral RNA was detected by RT-qPCR in order to phenotypically categorize animals into infected and not infected. In particular, 36 haplotypes were found, and their frequencies ranged from 0.13% to 35.15%. There were 62 resulting genotypes, three of which were associated with a putative resistance to the disease. Both the haplotypes and genotypes were inferred by PHASE v.2.1 software. To the best of our knowledge, this type of investigation was conducted for the first time on pig livestock distributed in different regions of Central Italy. Thus, the obtained findings may be considered very important since they add useful information about swine genetic background in relation to PRRS infection, from the perspective of adopting Marker-Assisted Selection (MAS) as a possible and alternative strategy to control this still widespread disease.
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Wilcock, Pete, Kiah M. Berg, Gustavo Cordero i Anthony Russo. "PSII-14 The Effect of Increasing Soluble Dietary Fiber and the Addition of a Stimbiotic in Gestation on Sow Productivity in a Prrs and Non-Prrs Challenge". Journal of Animal Science 101, Supplement_2 (28.10.2023): 290–91. http://dx.doi.org/10.1093/jas/skad341.331.
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Abstract This sow study evaluated the effect that sugar beet pulp (SBP), a soluble fiber source, and a stimbiotic (Signis; AB Vista) can have on sow productivity and health when fed in gestation. A stimbiotic is an additive that stimulates fiber-degrading microbiome, resulting in an increase in fiber fermentability even though the additive itself contributes limited small chain fatty acid production. A commercial gestation barn with two feeding lines was utilized to feed sows (n = 2173) two dietary treatments from d 30 to d 90; a corn, soy, soy hulls with (HSDF) or without (LSDF) SBP (50 kg/t) plus stimbiotic (100 g/t) and from d 90 to entering the farrowing pen the LSDF diet with or without a stimbiotic (100 g/day). Sows were fed 2.0 kg per day providing the following fiber intake levels/day, HDSF: TDF; 266g, SDF 55g, IDF, 211g and LDSF: TDF; 240g, SDF 40g, IDF, 200g. The following variables were measured; total born, live born, still born, mummies, pre-weaning mortality, pigs weaned per sow, prolapse rate and sow mortality/culls. A proportion of sows (n = 81) were moved to a GESTAL system for lactation and intake recorded per gestation treatment. During the study the farm became PRRS positive and any sows that farrowed while the farm was PRRS positive were identified as PRRS positive sows and included as a factor. Data were analyzed with ANOVA evaluating the effects of fiber, parity, and PRRS using the fit model platform in JMP16. Significant treatment means were separated using Student’s t-tests, with significance accepted at P ≤ 0.05. Results showed that for born alive there was an interaction between treatment and parity (P =0.02) and treatment and PRRS (P < 0.05) with the HDSF diet increasing born alive in parity 1 (+1.30 pigs; P = 0.03) and parity 2 (+0.71 pigs; P = 0.08) with no effect in parity 3+. HDSF improved born alive in non PRRS pigs (+1.01 pigs; P < 0.01) and had no effect in PRRS pigs. There was a tendency for HDSF program to reduce stillborn (8.8% v 7.4%; P = 0.09) but had no effect on mummies. There was a tendency to increase in pigs weaned with the HSDF treatment (9.15 v 8.86; P = 0.07) with no effect on sow mortality/culls. There was no effect of treatment on prolapses. On the sub-sample of sows recorded in lactation those sows (Parity 2+) that had been on the HSDF in gestation showed an increased ADFI (5.36 kg v 6.02 kg; P = 0.01) compared with the LSDF. It can be concluded that sow productivity can be improved by increasing the fermentative capacity of the gestation feed through the combination of adding a soluble fiber such as SBP along with a stimbiotic.
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ROLINEC, Michal, Daniel Bíro, Branislav Gálik, Milan Šimko i Miroslav Juráček. "IMMUNOGLOBULINS IN COLOSTRUM OF SOWS WITH PORCINE REPRODUCTIVE AND RESPIRATORY SYNDROME - PRRS". Journal of Central European Agriculture 13, nr 2 (2012): 301–8. http://dx.doi.org/10.5513/jcea01/13.2.1049.
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Graham, Simon P., Yuen-Ki Cheong, Summer Furniss, Emma Nixon, Joseph A. Smith, Xiuyi Yang, Rieke Fruengel i in. "Antiviral Efficacy of Metal and Metal Oxide Nanoparticles against the Porcine Reproductive and Respiratory Syndrome Virus". Nanomaterials 11, nr 8 (20.08.2021): 2120. http://dx.doi.org/10.3390/nano11082120.
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Porcine reproductive and respiratory syndrome viruses (PRRSV) are responsible for one of the most economically important diseases affecting the global pig industry. On-farm high-efficiency particulate air (HEPA) filtration systems can effectively reduce airborne transmission of PRRSV and the incidence of PRRS, but they are costly, and their adoption is limited. Therefore, there is a need for low-cost alternatives, such as antimicrobial filters impregnated with antiviral nanoparticles (AVNP). During the past 10 years, tailored intermetallic/multi-elemental AVNP compositions have demonstrated effective performance against human viruses. In this study, a panel of five AVNP was evaluated for viricidal activity against PRRSV. Three AVNP materials: AVNP2, copper nanoparticles (CuNP), and copper oxide nanoparticles (CuONP), were shown to exert a significant reduction (>99.99%) in virus titers at 1.0% (w/v) concentration. Among the three, CuNP was the most effective at lower concentrations. Further experiments revealed that AVNP generated significant reductions in viral titers within just 1.5 min. For an optimal reduction in viral titers, direct contact between viruses and AVNP was required. This was further explained by the inert nature of these AVNP, where only negligible leaching concentrations of Ag/Cu ions (0.06–4.06 ppm) were detected in AVNP supernatants. Real-time dynamic light scatting (DLS) and transmission electron microscopic (TEM) analyses suggested that the mono-dispersive hydrodynamic behavior of AVNPs may have enhanced their antiviral activity against PRRSV. Collectively, these data support the further evaluation of these AVNP as candidate nanoparticles for incorporation into antimicrobial air-filtration systems to reduce transmission of PRRSV and other airborne pathogens.
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Wang, Jingjing, Zhaohai Du, Xuehan Huo, Juan Zhou, Yu Chen, Jingxia Zhang, Ao Pan, Xiaoyang Wang, Furong Wang i Jun Zhang. "Genome-wide analysis of PRR gene family uncovers their roles in circadian rhythmic changes and response to drought stress in Gossypium hirsutum L." PeerJ 8 (25.09.2020): e9936. http://dx.doi.org/10.7717/peerj.9936.
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Background The circadian clock not only participates in regulating various stages of plant growth, development and metabolism, but confers plant environmental adaptability to stress such as drought. Pseudo-Response Regulators (PRRs) are important component of the central oscillator (the core of circadian clock) and play a significant role in plant photoperiod pathway. However, no systematical study about this gene family has been performed in cotton. Methods PRR genes were identified in diploid and tetraploid cotton using bioinformatics methods to investigate their homology, duplication and evolution relationship. Differential gene expression, KEGG enrichment analysis and qRT-PCR were conducted to analyze PRR gene expression patterns under diurnal changes and their response to drought stress. Results A total of 44 PRR family members were identified in four Gossypium species, with 16 in G. hirsutum, 10 in G. raimondii, and nine in G. barbadense as well as in G. arboreum. Phylogenetic analysis indicated that PRR proteins were divided into five subfamilies and whole genome duplication or segmental duplication contributed to the expansion of Gossypium PRR gene family. Gene structure analysis revealed that members in the same clade are similar, and multiple cis-elements related to light and drought stress response were enriched in the promoters of GhPRR genes. qRT-PCR results showed that GhPRR genes transcripts presented four expression peaks (6 h, 9 h, 12 h, 15 h) during 24 h and form obvious rhythmic expression trend. Transcriptome data with PEG treatment, along with qRT-PCR verification suggested that members of clade III (GhPRR5a, b, d) and clade V (GhPRR3a and GhPRR3c) may be involved in drought response. This study provides an insight into understanding the function of PRR genes in circadian rhythm and in response to drought stress in cotton.
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Liu, Wenguang, Bing Liu, Gege Zhang, Huixia Jia, Yang Zhang, Xitong Cen, Gaoyou Yao i Maoxian He. "Molecular and Functional Characterization of a Short-Type Peptidoglycan Recognition Protein, Ct-PGRP-S1 in the Giant Triton Snail Charonia tritonis". International Journal of Molecular Sciences 23, nr 19 (21.09.2022): 11062. http://dx.doi.org/10.3390/ijms231911062.
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Peptidoglycan recognition proteins (PGRPs) are a family of pattern recognition receptors (PRRs) involved in host antibacterial responses, and their functions have been characterized in most invertebrate and vertebrate animals. However, little information is available regarding the potential function of PGRPs in the giant triton snail Charonia tritonis. In this study, a short-type PGRP gene (termed Ct-PGRP-S1) was identified in C. tritonis. Ct-PGRP-S1 was predicted to contain several structural features known in PGRPs, including a typical PGRP domain (Amidase_2) and Src homology-3 (SH3) domain. The Ct-PGRP-S1 gene was constitutively expressed in all tissues examined except in proboscis, with the highest expression level observed in the liver. As a typical PRR, Ct-PGRP-S1 has an ability to degrade peptidoglycan (PGN) and was proven to have non-Zn2+-dependent amidase activity and antibacterial activity against Vibrioalginolyticus and Staphylococcus aureus. It is the first report to reveal the peptidoglycan recognition protein in C. tritonis, and these results suggest that peptidoglycan recognition protein Ct-PGRP-S1 is an important effector of C. tritonis that modulates bacterial infection resistance of V. alginolyticus and S. aureus, and this study may provide crucial basic data for the understanding of an innate immunity system of C. tritonis.
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Kyutoku, Fumiaki, Takashi Yokoyama i Katsuaki Sugiura. "Genetic Diversity and Epidemic Types of Porcine Reproductive and Respiratory Syndrome (PRRS) Virus in Japan from 2018 to 2020". Epidemiologia 3, nr 2 (3.06.2022): 285–96. http://dx.doi.org/10.3390/epidemiologia3020022.
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To clarify the genetic diversity of the porcine reproductive and respiratory syndrome virus (PRRSV) in Japan in recent years, we determined the nucleotide sequence of open reading frame 5 of 2482 PRRSV sequences obtained from samples collected from pigs between January 2018 and December 2020. As a result of molecular phylogenetic analysis, Cluster II represented the largest proportion (44.9–50.6%) throughout the study period, followed by Cluster IV (34.0–40.8%), Cluster III (7.8–12.1%), Cluster I (3.1–6.7%), and Cluster V (0.1–0.2%). The relative distributions between Clusters varied between geographic regions and between years: in 2018, Cluster II was the most prevalent in all regions. In 2019, Cluster II was dominant in the Hokkaido and Tohoku regions, while in other regions Cluster IV was dominant. In 2020, Cluster IV was dominant in the Kanto/Tosan and Kyushu/Okinawa regions, whilst in other regions Cluster II was predominant. Compared with a previous study, the proportions of genome sequences classified in Clusters II and IV significantly increased (p = 0.042 and 0.018, respectively) and those classified in Cluster III significantly decreased (p < 0.01). The widespread use of live attenuated vaccines using strains that belong to Cluster II might have accounted for these changes in the relative distribution between Clusters.
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Yoshino, Hironori, i Ikuo Kashiwakura. "Beneficial Effects Of Ionizing Radiation To Enhance Anti-Cancer Effects Of RIG-Like Receptor Stimulus". Blood 122, nr 21 (15.11.2013): 4721. http://dx.doi.org/10.1182/blood.v122.21.4721.4721.
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Introduction The immune system is composed of innate and adaptive immunity. Antigen presenting cells (APCs), such as macrophages and dendritic cells, serve as a link between innate and adaptive immunity. Furthermore, APCs express pattern recognition receptors (PRRs) recognize pathogen-associated molecular patterns. Retinoic acid-inducible gene-I (RIG-I)-like receptors [RLRs; RIG-I and melanoma differentiation-associated gene 5 (MDA5)] are a type of PRRs and sense virus-derived RNA or a synthetic analog of dsRNA polyinosinic-polycytidylic acid [poly(I:C)]. Although macrophages are resistant to ionizing radiation, it remains unclear whether radiation affects RLR expression in the macrophages. Therefore, the effects of ionizing radiation on the RLR expression in macrophages and the response against poly(I:C) were herein investigated. Additionally, the anti-cancer effects of poly(I:C) and the combination treatment of poly(I:C) with ionizing radiation was examined in human lung cancer A549 cells. Methods For preparation of human macrophage-like cells, the human acute monocytic leukemia THP1 cells were treated with phorbol 12-myristate 13-acetate and then differentiated into macrophage-like cells. To stimulate RLRs, poly(I:C)/LyoVecTM (InvivoGen), which is a complex between poly(I:C) and the transfection reagent LyoVecTM, was used on macrophage-like differentiated THP-1 cells. X-irradiation was performed with an X-ray generator at a dose rate of 102.0–104.0 cGy/min. The viable cells were counted by trypan blue exclusion assay. The expression of RLRs was analyzed by reverse transcription polymerase chain reaction (RT-PCR) or western blotting. The interferon (IFN)-β expression and tumor necrosis factor (TNF)-α concentration present in culture supernatants were analyzed by RT-PCR and enzyme-linked immunosorbent assay (ELISA), respectively. The cell death analysis was performed by fluorescein isothiocyanate labeled annexin V and propidium iodide (PI) staining and analyzed by flow cytometry. Results The effects of ionizing radiation on RLR expression were first investigated. Both non-irradiated and X-irradiated (1–10 Gy) macrophage-like cells expressed RIG-I and MDA-5, with no significant difference in expression levels. Next the response of macrophage-like cells to poly(I:C)/LyoVecTM (500 ng/ml) was examined. Although the expression of IFN-β was not observed in non-stimulated macrophage-like cells, the poly(I:C)/LyoVecTM-stimulated macrophage-like cells expressed IFN-β. In X-irradiated macrophage-like cells, IFN-β expression after poly(I:C)/LyoVecTM stimulation was comparable with that of non-irradiated cells. Similar to the IFN-β expression, no significant difference in concentration of TNF-α were observed after poly(I:C)/LyoVecTMstimulation in non-irradiated and irradiated cells. These results suggest that ionizing radiation did not affect RLR expression or the response against poly(I:C) in macrophages. We next investigated the anti-cancer effects of poly(I:C)/LyoVecTM against human lung cancer A549 cells. Treatment with poly(I:C)/LyoVecTM (500 and 1000 ng/ml) suppressed the A549 cell growth (approximately 70% and 80% inhibition, respectively). Furthermore, the treatment with poly(I:C)/LyoVecTM induced both annexin V(+)/PI(−) (early apoptotic cells) and annexin V(+)/PI(+) cells (late apoptotic/necrotic cells). Finally, the combination treatment of poly(I:C) with 2 Gy was tested. The cell growth suppressive effects of 2 Gy resulted in 30% inhibition, whereas the combination of 2 Gy with poly(I:C)/LyoVecTM (500 and 1000 ng/ml) resulted in 85% and 90% inhibition, respectively. Correspondingly, the proportion of early apoptotic cells and late apoptotic/necrotic cells were higher in the combination of 2 Gy with poly(I:C)/LyoVecTM compared with 2 Gy alone or poly(I:C)/LyoVecTMalone. These results suggest that poly(I:C) and ionizing radiation synergistically exhibit anti-cancer effects against A549 cells. Conclusion This study demonstrated that ionizing radiation synergistically acted with RLR stimulation in suppressing the growth of human lung cancer cells without affecting the expression of RLRs in macrophages. Therefore, the combination of radiation therapy with RLR stimulus is expected to be an effective cancer therapy. Disclosures: No relevant conflicts of interest to declare.
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Yeske, Paul. "41 Is there anything we can do to lessen the severity of PRRSv?" Journal of Animal Science 102, Supplement_2 (1.05.2024): 235–36. http://dx.doi.org/10.1093/jas/skae102.268.
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Abstract Veterinarians and the swine industry continue to battle PRRS virus (v), as demonstrated by both the MSHMP (Morrison Swine Health Monitoring Project) and the SDRS (Swine Disease Reporting System). Both systems depict a continued increase in PRRSv prevalence and percentage of positive cases. The PRRSv variants that are circulating today seem to be more severe, especially in the last 2 to 3 yr, with more significant health and production losses in the nursery and finishing phases for prolonged periods of time. The best thing that can be done is to avoid having the virus with a good bioexclusion plan and if sites become infected, a strong biocontainment plan to avoid contaminating more sites should be considered. Realizing that bioexclusion is not always possible and sites will become infected, there are methods to help to reduce the impact of the virus. With the concerns of recombination pushing the virus evolution along faster, more producers are working toward eliminating field viruses from their systems. The use of filtration both on sow farms and nurseries has become more common as well as an additional method of bioexclusion. In the face of an outbreak, obtaining an accurate diagnosis is still one of the most important things that we do. In farrow to wean farms, serum samples are generally used when clinical signs (e.g., abortions, off feed) are observed to target testing to specific animals that are more likely to allow for early detection. In nursery and finishing sites, the use of oral fluid has worked very well as a screening tool when there are clinical signs or problems within a group or flow. Fortunately, there are differential tests that can be used to identify field infection and vaccines strains (since most wean to finish pigs are vaccinated in pig dense areas). With severe strains, depopulation and repopulation strategies have become more popular as a control method, but the majority of herd would use a herd closure. However, the amount of time required for these closures is extending, which lengthens the impacts on sow farms. Among wean to finish systems, the use of depopulating and cleaning up the site is used most, with only routine down times. Anti-inflammatories can also be administered in the acute phase of the outbreak in addition to the use of antibiotics to control secondary bacterial infections, as well as providing a good environment to allow the pigs to better deal with the disease. It’s best to keep the virus out of the herd with a good bioexclusion program. However, when we do have outbreaks, anti-inflammatory and antibiotics to help the pigs through it are important to utilize as we work to eliminate the field virus from the herd.
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Modak, Tejashree H., i Marta Gomez-Chiarri. "Contrasting Immunomodulatory Effects of Probiotic and Pathogenic Bacteria on Eastern Oyster, Crassostrea Virginica, Larvae". Vaccines 8, nr 4 (6.10.2020): 588. http://dx.doi.org/10.3390/vaccines8040588.
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Several Vibrio spp. cause acute and severe mortality events in hatcheries where larvae of bivalve mollusks are reared, potentially leading to subsequent shortage of bivalve seed for the grow-out industry. In particular, strains of Vibrio coralliilyticus have been identified as a major cause of disease in Pacific, Crassostrea gigas, and eastern, C. virginica, oyster hatcheries in the USA of America. Probiotic bacteria are an inexpensive, practical, and natural method of disease control. Previous research shows that pretreatment of larval oysters with probiotic bacteria Bacillus pumilus RI06–95 (RI) and Phaeobacter inhibens S4 (S4) significantly decreases mortality caused by experimental challenge with the bacterial pathogen V. coralliilyticus RE22 (RE22). This study aims to characterize the immune response of 6–10-day-old eastern oyster larvae to experimental challenge with pathogen V. coralliilyticus RE22 and probionts RI and S4. Treatments included (a) pathogen and probiont exposure at a concentration of 5 × 104 CFU per mL (~2500 bacterial cells per larva) for a duration of 6 h, (b) probiont exposure at the same concentration for a duration of 24 h, and (c) probiont RI daily treatment of larvae in the hatchery for 4, 11, and 15 days. Differential gene expression analysis compared pathogen or probiotic-treated transcriptomes to unexposed controls. Probiotic and pathogen treatment led to upregulation of transcripts coding for several immune pattern recognition receptors (PRRs) involved in environmental sensing and detection of microbes in oyster larvae. Larval oyster responses to pathogen RE22 suggested suppression of expression of genes in immune signaling pathways (myd88, tak1, nkap), failure in upregulation of immune effector genes, high metabolic demand, and oxidative stress that potentially contributed to mortality. On the other hand, the transcriptomic response to probiotic bacteria RI and S4 suggested activation of immune signaling pathways and expression of immune effectors (e.g., Cv-spi2, mucins and perforin-2). These key features of the host immune response to probiotic bacteria were shared despite the length of probiotic exposure, probiotic species, and the type of environment in which exposures were conducted. This study suggests that pre-exposure of eastern oyster larvae to probiotics for 6–24 h prior to pathogenic challenge leads to a robust and effective immune response that may contribute to protecting larvae from subsequent challenge with V. coralliilyticus RE22. This research provides new insights into host-microbe interactions in larval oysters that could be applied in the management of vibriosis in bivalve hatcheries.
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Arepally, Gowthami M., Rui Qi, John Hollingsworth i Shayela Suvarna. "Determinants of PF4/Heparin Immunogenicity in a Murine Model of HIT." Blood 108, nr 11 (16.11.2006): 96. http://dx.doi.org/10.1182/blood.v108.11.96.96.
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Abstract Heparin-Induced Thrombocytopenia (HIT) is a life-threatening immune-mediated reaction caused by platelet factor 4 (PF4)/heparin complexes. In recent in vitro studies, we have shown that human and monoclonal HIT antibodies recognize and optimally bind to ultra-large complexes (ULC; MW>670kDa) formed at equimolar ratios of PF4 and heparin. We undertook the following studies to determine if the in vivo immunogenicity of PF4/heparin complexes showed similar requirements for stoichiometric molar ratios of PF4:heparin (PHR). Using a previously described murine immunization model, mice were immunized with antigen by retro-orbital injection daily for 5 days without adjuvant. Blood was collected at baseline and at weekly intervals for 4–6 weeks. Plasma was tested for antibody development by a murine PF4/heparin (mP+H) ELISA. To determine the effects of PHR on mP+H antibody production in vivo, PF4 was mixed with unfractionated heparin (UFH) at molar ratios of 2:1, 1:1 or 1:2 and injected into mice (n=5/cohort). Antibody levels (mean A450nm ± SD) of mice injected with PHR 2:1 (0.79 ± 0.77) were higher than mice injected with PHR 1:1 (0.23± 0. 2), or PHR 1:2 (0.09 ± 0.02, p<0.05 for PHR 2:1 v. PHR 1:2). Because in vitro studies showed that UFH was far more capable of generating ULCs when mixed with PF4 than low-molecular weight heparin (LMWH) or fondaparinux, mice (n=5/cohort) were injected with buffer, or mPF4 mixed with UFH, or enoxaparin (a LMWH) or the pentasaccharide, fondaparinux at doses that are therapeutically equivalent to doses given to humans. Antibody levels (mean A450nm ± SD) were significantly higher in mice injected with UFH (3.340 + 0.15) than in mice injected with buffer (0.0451 ± 0.013, p<0.001 for buffer v UFH), LMWH (0.8526 ± 0.44, p<0.001 for LMWH v UFH) or the pentasaccharide (0.80 ± 0.0.7, p<0.001 for pentasaccharide v UFH). Because PF4/heparin ULCs are composed of repeating structural antigenic units, we next investigated the role of toll-like receptors (TLRs), a family of pattern recognition receptors (PRRs) that recognize repeating structural determinants on microbes and foreign antigens. To determine if mP+H complexes engage TLRs, we injected mP+H into wild-type (WT, n=5) or mice deficient in MyD88 (n=7), an adaptor molecule critical for intracellular signaling by most TLRs. Antibody levels (mean A450nm ± SD) did not significantly differ between WT (1.5 ± 0.6) and MyD88 null mice (0.9 ± 0.5). In summary, these studies indicate that in vivo antibody responses to mP+H complexes are critically dependent on stoichiometric ratios favoring ULC formation and that immune activation by PF4/heparin is independent of MyD88 signaling pathways.
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Afzal, Muhammad Amin, Farrukh Azeem, Shumaila Afzal, Naila Afzal, Muhammad Rizwan, Hyojin Seo, Asad Ali Shah i Muhammad Amjad Nawaz. "Comparative Omics-Based Identification and Expression Analysis of a Two-Component System in Vigna radiata in Drought Stress". Agronomy 13, nr 4 (27.03.2023): 989. http://dx.doi.org/10.3390/agronomy13040989.
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Two-component system (TCS) genes regulate a wide range of biological activities in prokaryotes and eukaryotes, including plants. TCS plays an important role in cellular responses to external stimuli, such as biotic and abiotic factors. In plants, this system supports cell division, leaf senescence, stress response, chloroplast division, and nutrient signaling. There are three kinds of proteins responsible for the appropriate functioning of the TCS system: histidine kinases (HKs), histidine phosphotransfer proteins (HPs), and response regulators (RRs). The results of the current study revealed that Vigna radiata has 54 genes encoding potential TCS proteins, which were divided into three subgroups: 18 HKs, 9 HPs (seven true and two pseudos), and 27 RRs (8 type-A, 8 type-B, 3 type-C, and 8 PRRS). The anticipated TCS genes were widely dispersed across all eleven chromosomes and had family-specific intron/exon structures. After investigating TCS genes in a variety of plant species, we determined that Vigna HK (L)s, HPs, and RRs have closer evolutionary relationships with other legume genes. Gene duplication, including segmental and tandem types, is the most frequent source of gene family expansion. Multiple stress-related cis-elements were predicted in the promoter sequences of the VrTCS genes. RNA-seq data analysis demonstrated that VrTCS genes were expressed in clusters of upregulated and downregulated groups in response to drought stress. Moreover, these clusters were differentially expressed as early or late responses to drought stress. Real-time qPCR showed that VrHK2, VrHK3, VrPHYE, VrHP4.1, VrRR5.2, and VrRR10 genes were upregulated, while VrRR3 and VrHP6.1 genes were downregulated in response to drought stress. The current study highlights the architecture of V. radiata TCS and provides a robust framework for subsequent functional evaluation.
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Pavlakis, Nick, David Turner Ransom, David Wyld, Katrin Marie Sjoquist, Rebecca Asher, Val Gebski, Kate Wilson i in. "Australasian Gastrointestinal Trials Group (AGITG) CONTROL NET Study: Phase II study evaluating the activity of 177Lu-Octreotate peptide receptor radionuclide therapy (LuTate PRRT) and capecitabine, temozolomide CAPTEM)—First results for pancreas and updated midgut neuroendocrine tumors (pNETS, mNETS)." Journal of Clinical Oncology 38, nr 15_suppl (20.05.2020): 4608. http://dx.doi.org/10.1200/jco.2020.38.15_suppl.4608.
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4608 Background: CAPTEM is an accepted regimen for patients (pts) with advanced pNETs. Single agent 177Lu-Octreotate PRRT is now a standard of care for progressive WHO Grade (G) 1/2 mNETs. High activity was seen with LuTate/CAPTEM in a single arm Phase I/II trial. This study was undertaken to determine the relative activity of adding CAPTEM to LuTate PRRT in pts with mNETs and pNETs. Methods: Non-comparative randomised open label parallel group phase II trial with 2:1 randomisation to PRRT/CAPTEM (experimental arm) vs. PRRT (mNETs control) and CAPTEM (pNETS control). PRRT/CAPTEM: 7.8GBq LuTate day(D) 10, 8 weekly (wkly) x 4, with b.i.d. oral CAP 750mg/m2 D1-14 & TEM 75mg/m2D10-14, 8 wkly x 4; PRRT: 8 wkly x 4; CAPTEM 8 wkly x 4. Primary endpoint: Progression free survival (PFS). mNETS- at 15 months (mo) assuming 15mo PFS 66.4% in control arm, aiming for PFS ³ 80%; pNETS- at 12mo assuming 12mo PFS 60% in control arm, aiming for PFS ³ 75%. Secondary endpoints: Objective tumour response rate (complete or partial) (OTRR), clinical benefit rate (OTRR, stable disease) (CBR), toxicity, quality of life. Results: 75 pts enrolled (Dec 2015 – Nov 2018): mNETs 33 PRRT/CAPTEM and 14 PRRT; pNETS 19 PRRT/CAPTEM and 9 CAPTEM. mNETS: Median follow-up 35mo; 15mo PFS was 90% (95% CI: 73-97%) v 92% (95% CI: 57-99%); OTRR 31% vs 15%; and CBR 97% vs 92% for PRRT/CAPTEM v PRRT respectively. Treatment related adverse events (AEs): 24/32 PRRT/CAPTEM pts had at least one G3 event (75%) vs 5/13 (38%, PRRT); and 4/32 pts at least one G4 event (13%) v 1/13 (8%) respectively, mostly haematologic (haem). Only one patient failed to complete therapy (PRRT/CAPTEM). pNETS: Median follow-up 34mo; 12mo PFS was 76% (95% CI: 48-90%) v 67% (95% CI: 28-88%); OTRR 68% vs 33%; and CBR 100% vs 100% for PRRT/CAPTEM v CAPTEM respectively. Treatment related AEs: 5/18 PRRT/CAPTEM pts had at least one G3 event (28%) vs 3/9 (33%) CAPTEM; 3/18 pts at least one G4 event (17%) v 1/9 (11%) respectively. Conclusions: CAPTEM/PRRT is active, meeting its target landmark PFS for CAPTEM/PRRT (12mo pNETs; 15mo mNETs) with numerically greater OTRR in both pNETs and mNETs, but with more haem toxicity in mNETs. As activity was high in both control arms longer follow up is required to determine if the relative activity of PRRT/CAPTEM is sufficient to warrant Phase III evaluation. Clinical trial information: ACTRN12615000909527 .
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Reymond, Nicolas, Stéphanie Fabre, Eric Lecocq, José Adelaı̈de, Patrice Dubreuil i Marc Lopez. "Nectin4/PRR4, a New Afadin-associated Member of the Nectin Family That Trans-interacts with Nectin1/PRR1 through V Domain Interaction". Journal of Biological Chemistry 276, nr 46 (5.09.2001): 43205–15. http://dx.doi.org/10.1074/jbc.m103810200.
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Pavlakis, Nick, David Turner Ransom, David Wyld, Katrin Marie Sjoquist, Rebecca Asher, Val Gebski, Kate Wilson i in. "First results for Australasian Gastrointestinal Trials Group (AGITG) control net study: Phase II study of 177Lu-octreotate peptide receptor radionuclide therapy (LuTate PRRT) +/- capecitabine, temozolomide (CAPTEM) for midgut neuroendocrine tumors (mNETs)." Journal of Clinical Oncology 38, nr 4_suppl (1.02.2020): 604. http://dx.doi.org/10.1200/jco.2020.38.4_suppl.604.
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604 Background: Single agent 177Lu-octreotate peptide receptor radionuclide therapy is now a standard of care for progressive mNETS. High activity was seen with LuTate and concurrent CAPTEM chemotherapy in a single arm Phase I/II trial. This study was undertaken to determine the relative activity of adding CAPTEM to LuTate PRRT in patients with mNETs. Methods: Non-comparative randomised open label phase II trial of PRRT +/- CAPTEM in patients with mNETs, with 2:1 randomisation: PRRT /CAPTEM (experimental arm) vs. PRRT (control). PRRT /CAPTEM: 7.8GBq LuTate day(D) 10, 8 weekly (wkly) x 4, with b.i.d. oral CAP 750mg/m2 D1-14 & TEM 75mg/m2 D10-14, 8 wkly x 4, vs. PRRT 8 wkly x 4. Primary endpoint: progression free survival (PFS) at 15 months assuming 15 month PFS of 66.4% in the control arm, aiming for PFS rate > 80%; secondary endpoints: objective tumour response rate (complete or partial response) (OTRR), clinical benefit rate (complete or partial response, stable disease) (CBR), toxicity, and QOL. Results: 47 patients enrolled (Dec 2015 - Feb 2018): 33 PRRT/CAPTEM and 14 PRRT. Two patients withdrew prior to treatment. Patient characteristics were balanced except gender (female 58% vs. 14%). Two patients received 2 prior systemic regimens. After a median follow-up of 32 months, the 15 month PFS was 90% (95% CI: 73-97%) v 92% (95% CI: 57-99%); OTRR 25% vs 15%; and CBR 97% vs 92% for PRRT/CAPTEM v PRRT respectively. For treatment related adverse events 22/32 CAPTEM patients experienced one Grade 3 event (69%) vs 5/13 (38%, PRRT); 4/32 pts experienced one Grade 4 event (13%) v 1/13 (8%) respectively. Only one patient failed to complete therapy due to toxicity (PRRT/CAPTEM). Conclusions: This initial planned analysis demonstrates similarly high 15 month PFS for CAPTEM/PRRT relative to PRRT alone. OTRR is numerically higher but at the cost of greater toxicity. Longer follow up is required to determine if the activity of PRRT/CAPTEM is sufficient to warrant Phase III evaluation. Clinical trial information: ACTRN12615000909527.
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Mohammad, Amro H., Frédéric Couture, Isabelle Gamache, Owen Chen, Wissal El-Assaad, Nelly Abdel-Malak, Anna Kwiatkowska, William Muller, Robert Day i Jose G. Teodoro. "Cleavage of the V-ATPase associated prorenin receptor is mediated by PACE4 and is essential for growth of prostate cancer cells". PLOS ONE 18, nr 7 (18.07.2023): e0288622. http://dx.doi.org/10.1371/journal.pone.0288622.
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Phosphatase and tensin homolog (PTEN) mutation is common in prostate cancer during progression to metastatic and castration resistant forms. We previously reported that loss of PTEN function in prostate cancer leads to increased expression and secretion of the Prorenin Receptor (PRR) and its soluble processed form, the soluble Prorenin Receptor (sPRR). PRR is an essential factor required for proper assembly and activity of the vacuolar-ATPase (V-ATPase). The V-ATPase is a rotary proton pump required for the acidification of intracellular vesicles including endosomes and lysosomes. Acidic vesicles are involved in a wide range of cancer related pathways such as receptor mediated endocytosis, autophagy, and cell signalling. Full-length PRR is cleaved at a conserved consensus motif (R-X-X-R↓) by a member of the proprotein convertase family to generate sPRR, and a smaller C-terminal fragment, designated M8.9. It is unclear which convertase processes PRR in prostate cancer cells and how processing affects V-ATPase activity. In the current study we show that PRR is predominantly cleaved by PACE4, a proprotein convertase that has been previously implicated in prostate cancer. We further demonstrate that PTEN controls PRR processing in mouse tissue and controls PACE4 expression in prostate cancer cells. Furthermore, we demonstrate that PACE4 cleavage of PRR is needed for efficient V-ATPase activity and prostate cancer cell growth. Overall, our data highlight the importance of PACE4-mediated PRR processing in normal physiology and prostate cancer tumorigenesis.
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Cao, Zong-Xi, Pei-Rong Jiao, Yu-Mao Huang, Hong-Yang Qin, Liu-Wu Kong, Quan-Hui Pan, Yi-Min He i Gui-Hong Zhang. "Genetic diversity analysis of the ORF5 gene in porcine reproductive and respiratory syndrome virus samples from South China". Acta Veterinaria Hungarica 60, nr 1 (1.03.2012): 157–64. http://dx.doi.org/10.1556/avet.2012.013.
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To understand the genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV) in South China, we collected 231 clinical samples from pigs with suspected PRRSV infection in Guangdong between 2007 and 2009. We found that 74 of 231 samples were positive by RT-PCR. The PCR products of the ORF5 gene of 35 isolates from different farms were sequenced and their DNA sequences were compared to 23 other PRRSV isolates in the GenBank. We found that the nucleotide similarity among all South China isolates ranged from 87.6% to 100%, and all belonged to the North American genotype. Most of them were classified into subgenotype I, but the rest mapped to subgenotypes III, V or VI. Those in subgenotypes I and III were found to be highly variable in the primary neutralising epitope (PNE) with a specific amino acid mutation (F39/L39→I39), and a few isolates in subgenotypes I and III isolates also had a mutation at L41 (L41→S41). PRRSV isolates in subgenotypes III, V and VI had less potential glycosylation sites than those in subgenotype I. Our data contribute to the understanding of molecular variation of PRRSV in South China.
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Nguyen, Genevieve. "Renin, (pro)renin and receptor: an update". Clinical Science 120, nr 5 (19.11.2010): 169–78. http://dx.doi.org/10.1042/cs20100432.
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PRR [(pro)renin receptor] was named after its biological characteristics, namely the binding of renin and of its inactive precursor prorenin, that triggers intracellular signalling involving ERK (extracellular-signal-regulated kinase) 1/2. However the gene encoding for PRR is named ATP6ap2 (ATPase 6 accessory protein 2) because PRR was initially found as a truncated form co-purifying with V-ATPase (vacuolar H+-ATPase). There are now data showing that this interaction is not only physical, but also functional in the kidney and the heart. However, the newest and most fascinating development of PRR is its involvement in both the canonical Wnt/β-catenin and non-canonical Wnt/PCP (planar cell polarity) pathways, which are essential for adult and embryonic stem cell biology, embryonic development and disease, including cancer. In the Wnt/β-catenin pathway, it has been shown that PRR acts as an adaptor between the Wnt receptor LRP5/6 (low-density lipoprotein receptor-related protein 5/6) and Fz (frizzled) and that the proton gradient generated by the V-ATPase in endosomes is necessary for LRP5/6 phosphorylation and β-catenin activation. In the Wnt/PCP pathway, PRR binds to Fz and controls its asymetrical subcellular distribution and therefore the polarization of the cells in a plane of a tissue. These essential cellular functions of PRR are independent of renin and open new avenues on the pathophysiological role of PRR. The present review will summarize our knowledge of (pro)renin-dependent functions of PRR and will discuss the newly recognized functions of PRR related to the V-ATPase and to Wnt signalling.
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Indik, Stanislav, Lubomír Valíček, Dieter Klein i Jana Klánová. "Variations in the major envelope glycoprotein GP5 of Czech strains of porcine reproductive and respiratory syndrome virus". Journal of General Virology 81, nr 10 (1.10.2000): 2497–502. http://dx.doi.org/10.1099/0022-1317-81-10-2497.
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The major envelope glycoprotein genes (ORF5) of seven Czech isolates of porcine reproductive and respiratory syndrome virus (PRRSV) were amplified and their nucleotide sequences were determined. ORF5 displayed nucleotide and amino acid identities of 87·5–100% and 87·6–100%, respectively, among the isolates. In a phylogenetic tree, all European isolates were grouped in a genotype distinct from that of reference American strains (VR-2332, IAF-Klop). Among the European isolates, two different clades were identified. Two Czech isolates (V-501 and V-503) and Italian strain PRRSV 2156 fell into one clade. The remaining European strains comprised the second clade. Surprisingly, two separately clustered strains (V-501 and V-516) were isolated from the same herd. Additionally, the possible effect of in vitro cultivation on the nucleotide sequence was analysed. Nine point mutations in the ORF5 region resulted from 152 in vitro passages of the V-502 isolate in MARC-145 cells.
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Lopez, Marc, Mustapha Aoubala, François Jordier, Daniel Isnardon, Sophie Gomez i Patrice Dubreuil. "The Human Poliovirus Receptor Related 2 Protein Is a New Hematopoietic/Endothelial Homophilic Adhesion Molecule". Blood 92, nr 12 (15.12.1998): 4602–11. http://dx.doi.org/10.1182/blood.v92.12.4602.424k21_4602_4611.
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We have recently described Poliovirus Receptor Related 2 (PRR2), a new cell surface molecule homologous to the poliovirus receptor (PVR/CD155). Both molecules are transmembrane glycoproteins belonging to the Ig superfamily (IgSF). They contain 3 Ig domains of V, C2, and C2 types in their extracellular regions that share 51% aa identity. The PRR2 gene encodes two mRNA isoforms of 3.0 kb (hPRR2 [short form]) and 4.4 kb (hPRR2δ [long form]), both widely expressed in human tissues, including hematopoietic cells. To further characterize PRR2 expression during hematopoiesis and to analyze its function, we have developed a monoclonal antibody (MoAb) directed against its extracellular region (R2.477). PRR2 was expressed in 96% of the CD34+, 88% of the CD33+, and 95% of the CD14+ hematopoietic lineages and faintly in the CD41 compartment. Ectopic expression of both PRR2 cDNAs induced marked cell aggregation. A soluble chimeric receptor construct with the Fc fragment of human IgG1 (PRR2-Fc) as well as a fab fragment of the anti-PRR2 MoAb (R2.477) inhibit aggregation. PRR2-Fc binds specifically to PRR2-expressing cells. These results suggest that PRR2 is a homophilic adhesion receptor. PRR2 was also expressed at the surface of endothelial cells at the intercellular junctions of adjacent cells but not at the free cellular edges. Homophilic interactions are associated with dimerization of isoforms of PRR2 and lead to the tyrosine phosphorylation of PRR2δ. Altogether, these results suggest that homophilic properties of PRR2 could participate to the regulation of hematopoietic/endothelial cell functions.
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Matavelli, Luis C., Jiqian Huang i Helmy M. Siragy. "In vivo regulation of renal expression of (pro)renin receptor by a low-sodium diet". American Journal of Physiology-Renal Physiology 303, nr 12 (15.12.2012): F1652—F1657. http://dx.doi.org/10.1152/ajprenal.00204.2012.
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Effects of low salt (LS) on (pro)renin receptor (PRR) expression are not well established. We hypothesized that LS enhances renal PRR expression via the cGMP-protein kinase G (PKG) signaling pathway. Sprague-Dawley rats were fed a normal-salt (NS) or LS diet associated with intrarenal cortical administration of vehicle (V), the nitric oxide (NO) synthase inhibitor nitro-l-arginine methyl ester (l-NAME), the NO donor S-nitroso- N-acetyl-dl-penicillamine (SNAP), the cGMP analog 8-bromoguanosine (8-Br)-cGMP, the guanylyl cyclase inhibitor 1H-[1, 2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), or a PKG inhibitor (PKGi) for 6 days via osmotic minipump. We evaluated the effects of each treatment on renal interstitial fluid (RIF) levels of nitrate/nitrite and cGMP and renal PRR expression. There were no significant changes in blood pressure with any of the treatments. Urinary sodium excretion was significantly lower in rats given a LS diet. Compared with NS + V, RIF nitrate/nitrite and cGMP levels increased in LS + V rats. In NS groups, RIF nitrate/nitrite and cGMP levels did not change with l-NAME, ODQ, or PKGi and increased in response to SNAP. 8-Br-cGMP increased RIF cGMP but not RIF nitrate/nitrite. In LS groups, RIF nitrate/nitrite decreased with l-NAME and did not change with ODQ or PKGi whereas RIF cGMP decreased with l-NAME, ODQ, and PKGi. PRR mRNA and protein increased in LS + V. In NS rats, PRR mRNA and protein increased in response to 8-Br-GMP and were not affected by any of other treatments. In LS rats, PRR mRNA and protein decreased significantly in response to l-NAME, ODQ, and PKGi. We conclude that LS intake enhances renal expression of PRR via cGMP-PKG signaling pathway.
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Lopez, Marc, Mustapha Aoubala, François Jordier, Daniel Isnardon, Sophie Gomez i Patrice Dubreuil. "The Human Poliovirus Receptor Related 2 Protein Is a New Hematopoietic/Endothelial Homophilic Adhesion Molecule". Blood 92, nr 12 (15.12.1998): 4602–11. http://dx.doi.org/10.1182/blood.v92.12.4602.
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Abstract We have recently described Poliovirus Receptor Related 2 (PRR2), a new cell surface molecule homologous to the poliovirus receptor (PVR/CD155). Both molecules are transmembrane glycoproteins belonging to the Ig superfamily (IgSF). They contain 3 Ig domains of V, C2, and C2 types in their extracellular regions that share 51% aa identity. The PRR2 gene encodes two mRNA isoforms of 3.0 kb (hPRR2 [short form]) and 4.4 kb (hPRR2δ [long form]), both widely expressed in human tissues, including hematopoietic cells. To further characterize PRR2 expression during hematopoiesis and to analyze its function, we have developed a monoclonal antibody (MoAb) directed against its extracellular region (R2.477). PRR2 was expressed in 96% of the CD34+, 88% of the CD33+, and 95% of the CD14+ hematopoietic lineages and faintly in the CD41 compartment. Ectopic expression of both PRR2 cDNAs induced marked cell aggregation. A soluble chimeric receptor construct with the Fc fragment of human IgG1 (PRR2-Fc) as well as a fab fragment of the anti-PRR2 MoAb (R2.477) inhibit aggregation. PRR2-Fc binds specifically to PRR2-expressing cells. These results suggest that PRR2 is a homophilic adhesion receptor. PRR2 was also expressed at the surface of endothelial cells at the intercellular junctions of adjacent cells but not at the free cellular edges. Homophilic interactions are associated with dimerization of isoforms of PRR2 and lead to the tyrosine phosphorylation of PRR2δ. Altogether, these results suggest that homophilic properties of PRR2 could participate to the regulation of hematopoietic/endothelial cell functions.
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Cao, Xinran, Shiran Yu, Yuanyuan Wang, Min Yang, Jie Xiong, Haitao Yuan i Bo Dong. "Effects of the (Pro)renin Receptor on Cardiac Remodeling and Function in a Rat Alcoholic Cardiomyopathy Model via the PRR-ERK1/2-NOX4 Pathway". Oxidative Medicine and Cellular Longevity 2019 (13.03.2019): 1–13. http://dx.doi.org/10.1155/2019/4546975.
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Alcoholic cardiomyopathy (ACM) caused by alcohol consumption manifests mainly as by maladaptive myocardial function, which eventually leads to heart failure and causes serious public health problems. The (pro)renin receptor (PRR) is an important member of the local tissue renin-angiotensin system and plays a vital role in many cardiovascular diseases. However, the mechanism responsible for the effects of PRR on ACM remains unclear. The purpose of this study was to determine the role of PRR in myocardial fibrosis and the deterioration of cardiac function in alcoholic cardiomyopathy. Wistar rats were fed a liquid diet containing 9% v/v alcohol to establish an alcoholic cardiomyopathy model. Eight weeks later, rats were injected with 1×109v.g./100 μl of recombinant adenovirus containing EGFP (scramble-shRNA), PRR, and PRR-shRNA via the tail vein. Cardiac function was assessed by echocardiography. Cardiac histopathology was measured by Masson’s trichrome staining, immunohistochemical staining, and dihydroethidium staining. In addition, cardiac fibroblasts (CFs) were cultured to evaluate the effects of alcohol stimulation on the production of the extracellular matrix and their underlying mechanisms. Our results indicated that overexpression of PRR in rats with alcoholic cardiomyopathy exacerbates myocardial oxidative stress and myocardial fibrosis. Silencing of PRR expression with short hairpin RNA (shRNA) technology reversed the myocardial damage mediated by PRR. Additionally, PRR activated phosphorylation of ERK1/2 and increased NOX4-derived reactive oxygen species and collagen expression in CFs with alcohol stimulation. Administration of the ERK kinase inhibitor (PD98059) significantly reduced NOX4 protein expression and collagen production, which indicated that PRR increases collagen production primarily through the PRR-ERK1/2-NOX4 pathway in CFs. In conclusion, our study demonstrated that PRR induces myocardial fibrosis and deteriorates cardiac function through ROS from the PRR-ERK1/2-NOX4 pathway during ACM development.
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Nemec, James M., i Paweł Moskalik. "Four ‘Peculiar’ RRd stars observed by K2". Monthly Notices of the Royal Astronomical Society 507, nr 1 (9.07.2021): 781–802. http://dx.doi.org/10.1093/mnras/stab1929.
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ABSTRACT Four stars pulsating simultaneously with a dominant period PD ∈ (0.28, 0.39) d and an additional period PA ∈ (0.20, 0.27) d have been identified from among the more than 3000 RR Lyrae stars observed by the Kepler space telescope during NASA’s K2 Mission. All four stars are located in the direction of the Galactic Bulge and have period ratios, PA/PD, significantly smaller than those of most double-mode RR Lyrae (RRd) stars: PA/PD ∈ (0.694, 0.710) versus P1/P0 ∈ (0.726, 0.748). Three of the stars are faint (〈V〉 = 18–20 mag) and distant and are among the ‘peculiar’ RRd (pRRd) stars discovered by Prudil et al. (2017); the fourth star, EPIC 216764000 (= V1125 Sgr), is a newly discovered pRRd star several magnitudes brighter than the other three stars. In this paper, the high-precision long-cadence K2 photometry is analysed in detail and used to study the cycle-to-cycle light variations. The pulsational characteristics of pRRd stars are compared with those of ‘classical’ and ‘anomalous’ RRd (cRRd, aRRd) stars. The conclusion by Prudil et al. that pRRd stars form a separate group of double-mode pulsators and are not simply very short-period cRRd stars is confirmed. V1127 Aql and AH Cam are identified as other probable members of the class of pRRd stars.
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Fujimori, Takeshi, Daisuke Ogawa, Kenta Suzuki, Masaaki Kochi, Yuki Shibayama, Masaki Okada, Keisuke Miyake, Akira Nishiyama i Takashi Tamiya. "ET-04 MOLECULAR TARGETED THERAPY AGAINST (PRO)RENIN RECEPTOR FOR GLIOBLASTOMA". Neuro-Oncology Advances 1, Supplement_2 (grudzień 2019): ii8—ii9. http://dx.doi.org/10.1093/noajnl/vdz039.038.
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Abstract INTRODUCTION (Pro)renin receptor(PRR) is part of the Wnt receptor complex. Wnt/β-catenin signaling pathway (Wnt signaling) plays important role in pathogenesis and self-renewal of glioblastoma (GBM), or differentiation of glioma stem cell. We previously reported that PRR activate Wnt signaling, PRR expression correlated with malignancy of glioma, and treatment with PRR siRNA reduced the proliferative capacity. This time, we have developed monoclonal antibodies against PRR and examined their effects in GBM. MATERIAL AND METHODS We used GBM cell line (U251MG and U87MG) and primary human glioma stem cell line (MGG23). Glioma stem-like cells were cultured and isolated by neurosphere method from U251MG and U87MG. PRR antibody was made targeting the extracellular domain of the PRR with rat lymph node method. WST-1 assay or MTT assay were performed to determine the cell proliferation. Apoptosis was examined by FITC labeled annexin V and propidium iodide with flow cytometry. We analyzed molecules of Wnt signaling and stem cell markers with qRT-PCR. RESULTS We observed that PRR antibody significantly reduced cell proliferation, decreased sphere formation. Antibody suppressed cell adherent in stem-like cell. Flow cytometry showed that antibody induced apoptosis. Antibody inhibited Wnt signaling and stem cell markers. CONCLUSIONS PRR antibody reduced cell proliferation and induced apoptosis through Wnt signaling. PRR antibody also suppressed stemness. Our results demonstrated that PRR was a potential target for future glioma therapy.
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Abbas, Yazan M., Di Wu, Stephanie A. Bueler, Carol V. Robinson i John L. Rubinstein. "Structure of V-ATPase from the mammalian brain". Science 367, nr 6483 (12.03.2020): 1240–46. http://dx.doi.org/10.1126/science.aaz2924.
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In neurons, the loading of neurotransmitters into synaptic vesicles uses energy from proton-pumping vesicular- or vacuolar-type adenosine triphosphatases (V-ATPases). These membrane protein complexes possess numerous subunit isoforms, which complicates their analysis. We isolated homogeneous rat brain V-ATPase through its interaction with SidK, a Legionella pneumophila effector protein. Cryo–electron microscopy allowed the construction of an atomic model, defining the enzyme’s ATP:proton ratio as 3:10 and revealing a homolog of yeast subunit f in the membrane region, which we tentatively identify as RNAseK. The c ring encloses the transmembrane anchors for cleaved ATP6AP1/Ac45 and ATP6AP2/PRR, the latter of which is the (pro)renin receptor that, in other contexts, is involved in both Wnt signaling and the renin-angiotensin system that regulates blood pressure. This structure shows how ATP6AP1/Ac45 and ATP6AP2/PRR enable assembly of the enzyme’s catalytic and membrane regions.
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Yosypiv, Ihor V., Maria Luisa S. Sequeira-Lopez, Renfang Song i Alexandre De Goes Martini. "Stromal prorenin receptor is critical for normal kidney development". American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 316, nr 5 (1.05.2019): R640—R650. http://dx.doi.org/10.1152/ajpregu.00320.2018.
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Formation of the metanephric kidney requires coordinated interaction among the stroma, ureteric bud, and cap mesenchyme. The transcription factor Foxd1, a specific marker of renal stromal cells, is critical for normal kidney development. The prorenin receptor (PRR), a receptor for renin and prorenin, is also an accessory subunit of the vacuolar proton pump V-ATPase. Global loss of PRR is embryonically lethal in mice, indicating an essential role of the PRR in embryonic development. Here, we report that conditional deletion of the PRR in Foxd1+ stromal progenitors in mice ( cKO) results in neonatal mortality. The kidneys of surviving mice show reduced expression of stromal markers Foxd1 and Meis1 and a marked decrease in arterial and arteriolar development with the subsequent decreased number of glomeruli, expansion of Six2+ nephron progenitors, and delay in nephron differentiation. Intrarenal arteries and arterioles in cKO mice were fewer and thinner and showed a marked decrease in the expression of renin, suggesting a central role for the PRR in the development of renin-expressing cells, which in turn are essential for the proper formation of the renal arterial tree. We conclude that stromal PRR is crucial for the appropriate differentiation of the renal arterial tree, which in turn may restrict excessive expansion of nephron progenitors to promote a coordinated and proper morphogenesis of the nephrovascular structures of the mammalian kidney.
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Larrinaga, Gorka, Julio Calvete-Candenas, Jon Danel Solano-Iturri, Ana M. Martín, Angel Pueyo, Caroline E. Nunes-Xavier, Rafael Pulido, Juan F. Dorado, José I. López i Javier C. Angulo. "(Pro)renin Receptor Is a Novel Independent Prognostic Marker in Invasive Urothelial Carcinoma of the Bladder". Cancers 13, nr 22 (11.11.2021): 5642. http://dx.doi.org/10.3390/cancers13225642.
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(Pro)renin receptor (PRR) is being investigated in several malignancies as it activates pathogenic pathways that contribute to cell proliferation, immunosuppressive microenvironments, and acquisition of aggressive neoplastic phenotypes. Its implication in urothelial cancer (UC) has not been evaluated so far. We retrospectively evaluate the prognostic role of PRR expression in a series of patients with invasive UC treated with radical cystectomy and other clinical and histopathological parameters including p53, markers of immune-checkpoint inhibition, and basal and luminal phenotypes evaluated by tissue microarray. Cox regression analyses using stepwise selection evaluated candidate prognostic factors and disease-specific survival. PRR was expressed in 77.3% of the primary tumors and in 70% of positive lymph nodes. PRR expression correlated with age (p = 0.006) and was associated with lower preoperatively hemoglobin levels. No other statistical association was evidenced with clinical and pathological variables (gender, ASA score, Charlson comorbidity index, grade, pT, pN) or immunohistochemical expressions evaluated (CK20, GA-TA3, CK5/6, CD44, PD-L1, PD-1, B7-H3, VISTA, and p53). PRR expression in primary tumors was associated with worse survival (log-rank, p = 0.008). Cox regression revealed that PRR expression (HR 1.85, 95% CI 1.22–2.8), pT (HR 7.02, 95% CI 2.68–18.39), pN (HR 2.3, 95% CI 1.27–4.19), and p53 expression (HR 1.95, 95% CI 1.1–3.45) were independent prognostic factors in this series. In conclusion, we describe PRR protein and its prognostic role in invasive UC for the first time. Likely mechanisms involved are MAPK/ERK activation, Wnt/β-catenin signaling, and v-ATPAse function.
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Saito, Yoshinobu, Kenji Yokota, Kenji Yoshihara, Morihiro Saito, Jun Kuwano i Hidenobu Shiroishi. "Oxygen Reduction Electrode Properties of Pyrochlores Ln2Ru2O7-δ(Ln=Pr, Nd, Sm) in Aqueous Solutions". Key Engineering Materials 350 (październik 2007): 167–70. http://dx.doi.org/10.4028/www.scientific.net/kem.350.167.
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The activity of electrochemical oxygen reduction (EOR) for the pyrochlores Ln2Ru2-XO7-δ (Ln=Pr,Nd,Sm) [LnR] were examined in 0.1 M KOH aqueous solution at 70oC. The onset voltage (Vo) of the oxygen reduction current and the efficiency (E4) of 4-electron reduction of oxygen were evaluated by semi-steady state voltammetry with rotating ring-disk electrodes. PrR with the highest EOR activity showed Vo = ~ 0.85 V vs. reversible hydrogen electrode and E4 values above 80 %. Their Vo and E4 values show that LnR containing Ln with a smaller atomic number has a higher EOR activity, i.e. the order of the activity is PrR > NdR > SmR. This was in good agreement with that of the lattice parameters of LnR. These results indicate that the EOR activity of LnR depends on the kind and/or the size of the lanthanide metal ion on the A-site. PrR and NdR exhibited higher E4 values than known excellent Pb2Ru2O7-δ electrocatalyst containing toxic Pb.
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Harrison, Olivia, Haley Otott, Jianfa Bai, Vaughn Hamill, Aaron Singrey, Phillip C. Gauger, Marcelo Almeida i in. "67 Decontamination of a feed manufacturing environment following inoculation of porcine epidemic diarrhea virus, porcine reproductive and respiratory virus, and Seneca Valley virus 1". Journal of Animal Science 102, Supplement_2 (1.05.2024): 46–47. http://dx.doi.org/10.1093/jas/skae102.055.
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Abstract Feed mill decontamination is difficult because equipment is not designed to be cleaned with water. Alternate strategies may improve the ability of a mill to decontaminate in the event of viral contamination. The objective was to evaluate different decontamination strategies within a mill following the inoculation of porcine epidemic diarrhea virus (PEDV), porcine reproductive and respiratory syndrome virus (PRRSV), and Seneca Valley virus 1 (SVV1) contaminated feed. A batch of feed was inoculated with PEDV, PRRSV, and SVV1 and ran through a mixer, bucket elevator, corn cleaner, screw conveyor, distributor head, and spout. Following mill inoculation, decontamination strategies were implemented with environmental samples collected after each decontamination step. Decontamination strategies included: 1) complete facility decontamination including organic matter removal with hot water power washing, disinfection with 1% peroxygen (Virkon S, Lanxess, Germany) followed by a rinse, and a second disinfection with 5% sodium hypochlorite (Clorox, Oakland, CA) followed by a heating period for 48 h once the facility reached 60°C; 2) chlorine dioxide application (ProOxine AH, Bio-Cide International, Inc., Norman, OK); 3) organic matter removal using vacuums (Ridge Tool Company, Elyria, OH) and chlorine dioxide application; 4) heat up with portable electric heaters for exactly 48 h; and 5) organic matter removal and heat up with portable heaters for exactly 48 h. Samples were analyzed via triplex PCR. Cycle threshold and proportion PCR positive were analyzed using SAS GLIMMIX v 9.4 (SAS, Inc., Cary, NC). A swine bioassay was completed to determine the infectivity of samples following the final decontamination step of each treatment. A treatment × decontamination step × location interaction was observed (P < 0.05) for SVV1, where less RNA was detected post-treatment compared with pre-treatment following complete facility decontamination on surfaces including the mixer, corn cleaner, screw conveyor, and floor around the feed discharge (P < 0.05). There was no statistical difference between pre- and post-treatment samples for other combinations of decontamination treatment and location (P > 0.05), although numerical increases in Ct were generally observed following treatment. Across all treatments, the act of decontamination reduced detectable PEDV (P < 0.05) and PRRSV (P < 0.05) RNA when compared with samples immediately following inoculation, but complete facility decontamination was the only treatment where PEDV and PRRSV was non-detectable in all locations. Pigs given samples post-treatment showed no evidence of SVV1 or PEDV infection. Although PRRSV RNA was rarely found via PCR from samples collected within the mill, PRRSV infection was observed in pigs given the chlorine dioxide with and without organic matter removal treatments and the organic matter removal plus heat treatment. Overall, all treatments reduced detectable RNA for all viruses between the inoculation step and the final decontamination step; however, PRRSV particles remained infectious following decontamination regardless of PCR results in a swine bioassay.
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Miller, Benjamin F., Jill A. Fattor, Kevin A. Jacobs, Michael A. Horning, Sang-Hoon Suh, Franco Navazio i George A. Brooks. "Metabolic and cardiorespiratory responses to “the lactate clamp”". American Journal of Physiology-Endocrinology and Metabolism 283, nr 5 (1.11.2002): E889—E898. http://dx.doi.org/10.1152/ajpendo.00266.2002.
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To evaluate the hypothesis that precursor supply limits gluconeogenesis (GNG) during exercise, we examined training-induced changes in glucose kinetics [rates of appearance (Ra) and disappearance (Rd)], oxidation (Rox), and recycling (Rr) with an exogenous lactate infusion to 3.5–4.0 mM during rest and to pretraining 65% peak O2 consumption (V˙o 2 peak) levels during exercise. Control and clamped trials (LC) were performed at rest pre- (PRR, PRR-LC) and posttraining (POR, POR-LC) and during exercise pre- (PREX) and posttraining at absolute (POAB, POAB-LC) and relative (PORL, PORL-LC) intensities. Glucose Rrwas not different in any rest or exercise condition. Glucose Ra did not differ as a result of LC. Glucose Rox was significantly decreased with LC at POR (0.38 ± 0.03 vs. 0.56 ± 0.04 mg · kg−1 · min−1) and POAB (3.82 ± 0.51 vs. 5.0 ± 0.62 mg · kg−1 · min−1). Percent glucose Rd oxidized decreased with all LC except PORL-LC (PRR, 32%; PRR-LC, 22%; POR, 27%; POR-LC, 20%; POAB, 95%; POAB-LC, 77%), which resulted in a significant increase in oxidation from alternative carbohydrate (CHO) sources at rest and POAB. We conclude that 1) increased arterial [lactate] did not increase glucose Rrmeasured during rest or exercise after training, 2) glucose disposal or production did not change with increased precursor supply, and 3) infusion of exogenous CHO in the form of lactate resulted in the decrease of glucose Rox.
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Chen, Kaiyu, Siyuan Zhang, Yina Shao, Ming Guo, Weiwei Zhang i Chenghua Li. "A unique NLRC4 receptor from echinoderms mediates Vibrio phagocytosis via rearrangement of the cytoskeleton and polymerization of F-actin". PLOS Pathogens 17, nr 12 (13.12.2021): e1010145. http://dx.doi.org/10.1371/journal.ppat.1010145.
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Many members of the nucleotide-binding and oligomerization domain (NACHT)- and leucine-rich-repeat-containing protein (NLR) family play crucial roles in pathogen recognition and innate immune response regulation. In our previous work, a unique and Vibrio splendidus-inducible NLRC4 receptor comprising Ig and NACHT domains was identified from the sea cucumber Apostichopus japonicus, and this receptor lacked the CARD and LRR domains that are typical of common cytoplasmic NLRs. To better understand the functional role of AjNLRC4, we confirmed that AjNLRC4 was a bona fide membrane PRR with two transmembrane structures. AjNLRC4 was able to directly bind microbes and polysaccharides via its extracellular Ig domain and agglutinate a variety of microbes in a Ca2+-dependent manner. Knockdown of AjNLRC4 by RNA interference and blockade of AjNLRC4 by antibodies in coelomocytes both could significantly inhibit the phagocytic activity and elimination of V. splendidus. Conversely, overexpression of AjNLRC4 enhanced the phagocytic activity of V. splendidus, and this effect could be specifically blocked by treatment with the actin-mediated endocytosis inhibitor cytochalasin D but not other endocytosis inhibitors. Moreover, AjNLRC4-mediated phagocytic activity was dependent on the interaction between the intracellular domain of AjNLRC4 and the β-actin protein and further regulated the Arp2/3 complex to mediate the rearrangement of the cytoskeleton and the polymerization of F-actin. V. splendidus was found to be colocalized with lysosomes in coelomocytes, and the bacterial quantities were increased after injection of chloroquine, a lysosome inhibitor. Collectively, these results suggested that AjNLRC4 served as a novel membrane PRR in mediating coelomocyte phagocytosis and further clearing intracellular Vibrio through the AjNLRC4-β-actin-Arp2/3 complex-lysosome pathway.
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Cocchi, Francesca, Laura Menotti, Prisco Mirandola, Marc Lopez i Gabriella Campadelli-Fiume. "The Ectodomain of a Novel Member of the Immunoglobulin Subfamily Related to the Poliovirus Receptor Has the Attributes of a Bona Fide Receptor for Herpes Simplex Virus Types 1 and 2 in Human Cells". Journal of Virology 72, nr 12 (1.12.1998): 9992–10002. http://dx.doi.org/10.1128/jvi.72.12.9992-10002.1998.
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ABSTRACT We report on the functional cloning of a hitherto unknown member of the immunoglobulin (Ig) superfamily selected for its ability to confer susceptibility to herpes simplex virus (HSV) infection on a highly resistant cell line (J1.1-2 cells), derived by exposure of BHKtk− cells to a recombinant HSV-1 expressing tumor necrosis factor alpha (TNF-α). The sequence of herpesvirus Ig-like receptor (HIgR) predicts a transmembrane protein with an ectodomain consisting of three cysteine-bracketed domains, one V-like and two C-like. HIgR shares its ectodomain with and appears to be an alternative splice variant of the previously described protein PRR-1 (poliovirus receptor-related protein). Both HIgR and PRR-1 conferred on J1.1-2 cells susceptibility to HSV-1, HSV-2, and bovine herpesvirus 1. The viral ligand of HIgR and PRR-1 is glycoprotein D, a constituent of the virion envelope long known to mediate viral entry into cells through interaction with cellular receptor molecules. Recently, PRR-1, renamed HveC (herpesvirus entry mediator C), and the related PRR-2, renamed HveB, were reported to mediate the entry of HSV-1, HSV-2, and bovine herpesvirus 1, and the homologous poliovirus receptor was reported to mediate the entry of pseudorabies virus (R. J. Geraghty, C. Krummenacher, G. H. Cohen, R. J. Eisenberg, and P. G. Spear, Science 280:1618–1620, 1998; M. S. Warner, R. J. Geraghty, W. M. Martinez, R. I. Montgomery, J. C. Whitbeck, R. Xu, R. J. Eisenberg, G. H. Cohen, and P. G. Spear, Virology 246:179–189, 1998). Here we further show that HIgR or PRR-1 proteins detected by using a monoclonal antibody to PRR-1 are widely distributed among human cell lines susceptible to HSV infection and commonly used for HSV studies. The monoclonal antibody neutralized virion infectivity in cells transfected with HIgR or PRR-1 cDNA, as well as in the human cell lines, indicating a direct interaction of virions with the receptor molecule, and preliminarily mapping this function to the ectodomain of HIgR and PRR-1. Northern blot analysis showed that HIgR or PRR-1 mRNAs were expressed in human tissues, with the highest expression being detected in nervous system samples. HIgR adds a novel member to the cluster of Ig superfamily members able to mediate the entry of alphaherpesviruses into cells. The wide distribution of HIgR or PRR-1 proteins among human cell lines susceptible to HSV infection, coupled with the neutralizing activity of the antibody in the same cells, provides direct demonstration of the actual use of this cluster of molecules as HSV-1 and HSV-2 entry receptors in human cell lines. The high level of expression in samples from nervous system makes the use of these proteins in human tissues very likely. This cluster of molecules may therefore be considered to constitute bona fide receptors for HSV-1 and HSV-2.
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Shi, Xiaoli, Sandrine Opi, Adrien Lugari, Audrey Restouin, Thibault Coursindel, Isabelle Parrot, Javier Perez i in. "Identification and biophysical assessment of the molecular recognition mechanisms between the human haemopoietic cell kinase Src homology domain 3 and ALG-2-interacting protein X". Biochemical Journal 431, nr 1 (14.09.2010): 93–102. http://dx.doi.org/10.1042/bj20100314.
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SFKs (Src family kinases) are central regulators of many signalling pathways. Their functions are tightly regulated through SH (Src homology) domain-mediated protein–protein interactions. A yeast two-hybrid screen using SH3 domains as bait identified Alix [ALG-2 (apoptosis-linked gene 2)-interacting protein X] as a novel Hck (haemopoietic cell kinase) SH3 domain interactor. The Alix–Hck-SH3 interaction was confirmed in vitro by a GST (glutathione transferase) pull-down assay and in intact cells by a mammalian two-hybrid assay. Furthermore, the interaction was demonstrated to be biologically relevant in cells. Through biophysical experiments, we then identified the PRR (proline-rich region) motif of Alix that binds Hck-SH3 and determined a dissociation constant of 34.5 μM. Heteronuclear NMR spectroscopy experiments were used to map the Hck-SH3 residues that interact with an ALIX construct containing the V and PRR domains or with the minimum identified interacting motif. Finally, SAXS (small-angle X-ray scattering) analysis showed that the N-terminal PRR of Alix is unfolded, at least before Hck-SH3 recognition. These results indicate that residues outside the canonical PxxP motif of Alix enhance its affinity and selectivity towards Hck-SH3. The structural framework of the Hck–Alix interaction will help to clarify how Hck and Alix assist during virus budding and cell-surface receptor regulation.
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Xiong, Lei, Ji-Ung Jung, Hao-Han Guo, Jin-Xiu Pan, Xiang-Dong Sun, Lin Mei i Wen-Cheng Xiong. "Osteoblastic Lrp4 promotes osteoclastogenesis by regulating ATP release and adenosine-A2AR signaling". Journal of Cell Biology 216, nr 3 (13.02.2017): 761–78. http://dx.doi.org/10.1083/jcb.201608002.
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Bone homeostasis depends on the functional balance of osteoblasts (OBs) and osteoclasts (OCs). Lrp4 is a transmembrane protein that is mutated in patients with high bone mass. Loss of Lrp4 in OB-lineage cells increases bone mass by elevating bone formation by OBs and reducing bone resorption by OCs. However, it is unclear how Lrp4 deficiency in OBs impairs osteoclastogenesis. Here, we provide evidence that loss of Lrp4 in the OB lineage stabilizes the prorenin receptor (PRR) and increases PRR/V-ATPase–driven ATP release, thereby enhancing the production of the ATP derivative adenosine. Both pharmacological and genetic inhibition of adenosine-2A receptor (A2AR) in culture and Lrp4 mutant mice diminishes the osteoclastogenic deficit and reduces trabecular bone mass. Furthermore, elevated adenosine-A2AR signaling reduces receptor activator of nuclear factor κB (RANK)–mediated osteoclastogenesis. Collectively, these results identify a mechanism by which osteoblastic Lrp4 controls osteoclastogenesis, reveal a cross talk between A2AR and RANK signaling in osteoclastogenesis, and uncover an unrecognized pathophysiological mechanism of high-bone-mass disorders.
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Sahu, Prince, Mahendra Singh, Rakesh Pandey, Mukesh Kumar Mishra, Akhilesh Kumar Singh, Bhupendra Kumar Singh, Surendra Kumar Singh i in. "Screening of Comprehensive Panel of Cultivated and Wild Vigna Species for Resistance to Pulse Beetle, Callosobruchus chinensis L." Biology 12, nr 6 (27.05.2023): 781. http://dx.doi.org/10.3390/biology12060781.
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Pulses are a key source of dietary proteins in human nutrition. Despite several efforts to increase the production, various constraints, such as biotic and abiotic factors, threaten pulse production by various means. Bruchids (Callosobruchus spp.) are the serious issue of concern, particularly in storage conditions. Understanding host–plant resistance at morphological, biochemical and molecular levels is the best way to minimize yield losses. The 117 mungbean (Vigna radiata L. Wilczek) genotypes, including endemic wild relatives, were screened for resistance against Callosobruchus chinensis; among them, two genotypes, PRR 2008-2 and PRR 2008-2-sel, which belong to V. umbellata (Thumb.), were identified as highly resistant. The expression of antioxidants in susceptible and resistant genotypes revealed that the activity of phenylalanine ammonia lyase (PAL) was upregulated in the highly resistant wild Vigna species and lower in the cultivated susceptible genotypes, along with other biomarkers. Further, the SCoT-based genotyping revealed SCoT-30 (200 bp), SCoT-31 (1200 bp) and SCoT-32 (300 bp) as unique amplicons, which might be useful for developing the novel ricebean-based SCAR markers to accelerate the molecular breeding programme.
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Harrison, Olivia, Catherine Grace Elijah, Allison K. Blomme, Haley Ottot, Jianfa Bai, Elizabeth Poulsen-Porter, Jason C. Woodworth, Chad B. Paulk, Jordan T. Gebhardt i Cassandra K. Jones. "53 Evaluating the Efficacy of Boot Baths with Wet and Dry Disinfectants for Porcine Epidemic Diarrhea Virus and Porcine Reproductive and Respiratory Syndrome Virus". Journal of Animal Science 100, Supplement_2 (12.04.2022): 17–18. http://dx.doi.org/10.1093/jas/skac064.029.
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Abstract Maintaining biosecurity between barns is challenging. Boot baths, either wet or dry, can be implemented to limit pathogen spread. The objective was to evaluate the efficacy of boot baths using wet or dry disinfectants for porcine epidemic diarrhea virus (PEDV) and porcine reproductive and respiratory syndrome virus (PRRSV). Treatments included 1) control, 2) wet disinfectant (Synergize, Neogen, Lexington, KY), and 3) dry disinfectant (Traffic C.O.P., PSP, LLC, Rainsville, AL). Prior to disinfection, 0.5 mL of both PRRSV (~1×105 TCID50/mL) and PEDV (~1×105 TCID50/mL) was placed onto a new boot with a layer of autoclaved corn dust and allowed to dry for 15 minutes. After the mixture dried, the boot was put on and stepped into its respective boot bath. After 3 seconds, the boot was lifted out of the bath and stepped onto a stainless-steel coupon to simulate walking through a facility. Both boot and coupon were allowed to dry for 1 minute before swabs were taken from both surfaces. Samples were analyzed in a duplex PCR at the Kansas State Veterinary Diagnostic Laboratory. Cycle threshold values were analyzed using SAS GLIMMIX v 9.4 (SAS, Inc., Cary, NC). There was no evidence of a treatment×surface×virus interaction (P > 0.10). The interaction between treatment×surface impacted (P < 0.05) the quantity of detectable RNA. The control had greater concentration of virus on the coupon than the boot. The reverse was true for boots treated with wet disinfectant, where the boot had a greater concentration of virus than the coupon. Treatment×virus also impacted detectable RNA (P < 0.05), where wet and control boots had greater quantities of PEDV RNA than PRRSV. There was no detectable virus when dry disinfectant was used. For this trial, dry disinfectant was the most efficacious in reducing the viral RNA on both boots and subsequent surfaces; however, further research in commercial settings is warranted.
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Zhang, Tong, Yang Liu, Ke Yang, Ming Tang, Xiang Yu i Fei Yu. "Hydromechanical Coupling Characteristics of the Fractured Sandstone under Cyclic Loading-Unloading". Geofluids 2020 (30.10.2020): 1–12. http://dx.doi.org/10.1155/2020/8811003.
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The mechanical and hydraulic properties of rock mass play a crucial role in underground engineering. To study the effect of hydraulic pressure, confining pressure, and axial cyclic loading-unloading on variation of the deformation and permeability in fractured rock mass, the coupling triaxial experiment of sandstone was conducted. The concept of permeability recovery rate (PRR) and permeability enhancement reduction rate (PERR) was proposed to characterize the change in permeability. The results show that the permeability of fractured sandstone quadratically varies with the change of hydraulic pressure and confining stress. In detail, the permeability decreases with the decrease of hydraulic pressure and increases with the decrease of confining stress, respectively. Compared with the single-fracture permeability, the double-fracture permeability is more sensitive to the change of hydraulic pressure. Furthermore, the permeability of fractured sandstone is more dependent on the hydraulic pressure than the confining stress. With the performance of axial cyclic loading-unloading, the permeability spirals down, and both the axial and radial residual strains quadratically evolve. Following the first axial cyclic loading-unloading, an obvious deformation memory phenomenon characterized by a parallelogram shape in axial stress-strain curves was observed for the sandstone. The cumulative PRR of 85%-95% was maintained in double-fracture sandstone. On the contrary, a fluctuation of cumulative PRR characterized by “V shape” was observed for single-fracture sandstone. The enhancement effect of axial cyclic loading on the permeability was characterized by the decrease of PERR for double-fracture sandstone and increase of PERR with a greater gradient for single-fracture sandstone.
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Eddicks, Matthias, Lina Eddicks, Julia Stadler, Walter Hermanns i Mathias Ritzmann. "Der Porcine Respiratory Disease Complex (PRDC) – eine klinische Übersicht". Tierärztliche Praxis Ausgabe G: Großtiere / Nutztiere 49, nr 02 (kwiecień 2021): 120–32. http://dx.doi.org/10.1055/a-1403-1976.
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ZusammenfassungDer Porcine Respiratory Disease Complex (PRDC) beschreibt eine klinische Kondition, die sich in Form einer häufig therapieresistenten Atemwegsinfektion bei Mastschweinen manifestiert. Die multifaktorielle Ätiologie beinhaltet infektiöse und nicht infektiöse Faktoren. Bei Entstehung und Verlauf des PRDC spielen neben Management und Hygiene v. a. virale und bakterielle Erreger eine bedeutende Rolle. Das Virus des Porzinen Reproduktiven und Respiratorischen Syndroms (PRRSV), das porzine Circovirus Typ 2 (PCV2), Influenza-A-Virus (IAV) und Mycoplasma (M.) hyopneumoniae stellen die relevantesten Erreger dar. Das klinische Bild und die zugrundeliegenden pathomorphologischen Veränderungen können je nach Erregerbeteiligung variieren. Die Komplexität des PRDC erschwert die Diagnose und auch die Prävention auf Bestandsebene. Der Übersichtsartikel gibt einen Einblick in die Pathomorphologie, Pathogenese sowie Inter-Erreger-Interaktionen und zielt darauf ab, praktizierende Tierärztinnen und Tierärzte bei der Diagnose, Befundinterpretation und Prävention des PRDC zu unterstützen.
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Roper, D. A., F. N. Scenna, A. M. Saxton, J. L. Edwards, N. R. Rohrbach i F. N. Schrick. "144 PREGNANCY RETENTION OF BOVINE RECIPIENTS FOLLOWING TRANSFER OF EMBRYOS EXPOSED TO A PROSTAGLANDIN2ALPHA RECEPTOR ANTAGONIST DURING COLLECTION". Reproduction, Fertility and Development 21, nr 1 (2009): 171. http://dx.doi.org/10.1071/rdv21n1ab144.
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Increasing efficiency and success of MOET continues to be a goal of researchers and practitioners. Although numerous studies report success in establishing pregnancies, fewer evaluate term development and report number of live calves born. In a previous study, Scenna FN et al. (2008 Reprod. Fertil. Dev. 20, 154) exposed embryos to 3 different medium treatments while being collected from superovulated beef donors on Day 7. Medium treatments consisted of a commercially available medium plus 1 mL of DMSO (Control), commercial medium plus 100 nm of AL-8810 (AL100), or a commercial medium plus 1000 nm of AL-8810 (AL1000). Embryos were evaluated for grade and stage according to IETS guidelines. Embryos (n = 1734 at 6 locations across 13 replicates) were transferred (fresh or frozen in ethylene glycol) by 4 experienced technicians. Pregnancy rates were determined by ultrasonography 28 to 35 days after transfer and were increased in recipients receiving embryos collected in media containing AL100 (65%) and AL1000 (60%) compared with Control (50%; P < 0.05) as previously reported. As a continuum of this research interest, pregnancy retention rates (PRR) of recipient animals receiving embryos exposed to a prostaglandin F2α receptor antagonist (AL100 or AL1000) during collection or serving as nontreated controls were determined. Pregnancy retention rate was defined as the percentage of animals calving that were diagnosed pregnant at 28 to 35 days after transfer. From this pool of recipient animals, calving information was available to date on 494 confirmed pregnancies (presence of an embryo with a heartbeat). Data were obtained on 192 Control, 108 AL100, and 194 AL1000 recipients and analyzed using generalized mixed model analysis of variance (Glimmix) in SAS (SAS Institute, Cary, NC). Similar to initial pregnancy rates, PRR did not differ between AL100 and AL1000 (89 ± 0.04% and 90 ± 0.03%, respectively; P > 0.10); therefore, these groups were combined (AL) for subsequent analysis of PRR. Regardless of treatment, overall PRR were 88 ± 0.01% for fresh and 85 ± 0.03% for frozen embryos (P > 0.10). Additionally, gestation length (days) was similar between recipient animals receiving Control (282 ± 1.19), AL100 (281 ± 1.30) or AL1000 (281 ± 1.17; P > 0.10). However, addition of AL to collection medium of embryos increased PRR compared with Control (90 ± 0.03% v. 83 ± 0.03%, respectively; P = 0.06). Currently, an interaction was not noted between freezing method and media treatment (P > 0.10). Although collection of data continues, addition of a prostaglandin F2α receptor antagonist to the collection medium of embryos improves PRR of recipient animals without altering gestation length.
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Gosewisch, Astrid, Harun Ilhan, Lena Vomacka i Guido Böning. "Dosimetrie bei der Radionuklidtherapie mit Lu-177". Der Nuklearmediziner 41, nr 01 (marzec 2018): 69–80. http://dx.doi.org/10.1055/s-0043-122171.
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ZusammenfassungDie Radioligandentherapie (RLT) mit radioaktiv markierten PSMA-Liganden (z. B. Lu-177-PSMA-617) und die Peptid-Rezeptor-Radionuklidtherapie (PRRT) mit radioaktiv markierten Somatostatin-Analoga (z. B. Lu-177-DOTATATE) haben sich über die letzten Jahre als vielversprechende Therapieoptionen bei metastasierten, kastrationsresistenten Prostatakarzinomen bzw. inoperablen oder metastasierten neuroendokrinen Tumoren entwickelt. Das Theragnostiknuklid Lu-177 weist dabei neben der Therapie-relevanten Beta-Minus-Komponente eine, auch für die Bildgebung gut nutzbare, Gammaemission auf, die eine direkte quantitative Bildgebung und Dosimetrie der Radiopharmakonverteilung möglich macht. Die Patienten-spezifische 3D-Dosimetrie auf Basis der quantitativen Lu-177-Bildgebung ist dabei der Schlüssel hin zur individualisierten Therapiekontrolle und -planung in der RLT und PRRT, in denen bisher vornehmlich noch standardisierte Aktivitätsmengen verabreicht werden. Jedoch ist die Ermittlung robuster Dosiswerte klinisch aufwändig, sowohl bez. des Messaufwands als auch hinsichtlich der Auswertung der Daten, und die starke Variabilität der Dosiswerte und deren Abhängigkeit von der verwendeten Methodik erschweren die Etablierung Therapie-spezifischer Dosis-Wirkungsbeziehungen. Gerade der Vergleich von Dosimetriedaten zwischen den Kliniken erfordert eine systematische und vereinheitlichte Dosimetrie mit akzeptablem klinischem Aufwand. Die Genauigkeit der Dosimetrie ist derzeit v. a. limitiert durch Effekte der Bildgebung. Zusätzliche Fehler können sich bspw. durch eine ungünstige Wahl der gemessenen Datenpunkte, Bewegungsartefakte, Überlagerungseffekte in der planaren Bildgebung oder durch eine nicht vollständig klinisch durchführbare Definition der Zielregion (z. B. Knochenmark) ergeben. Die 3D-Dosimetrie, unter Berücksichtigung zusätzlicher physikalischer oder biologischer Parameter (z. B. Dosisrate), kann in der Radionuklidtherapie einen wichtigen Beitrag zur Etablierung von Dosis-Wirkungsbeziehungen liefern, allerdings ist die klinisch-physiologische Interpretation der 3D-Dosisverteilungen derzeit noch stark limitiert durch Bildartefakte und Bildrauschen in den zur Dosimetrie eingesetzten SPECT-Bildern.
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Gorga, Marilena, Alina-Mihaela Soare, Ana-Maria Gherghe, Rucsandra Dobrota, Cristina Mambet i Carina Mihai. "A PILOT STUDY ON SOLUBLE (PRO)RENIN RECEPTOR IN SYSTEMIC SCLEROSIS". Romanian Journal of Rheumatology 24, nr 2 (30.06.2015): 108–13. http://dx.doi.org/10.37897/rjr.2015.2.7.
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Background. Wnt signaling is involved in fibrosis, but the mechanisms of cross-talk with other pathways, like TGF-β, are not fully understood. (Pro)renin receptor (PRR) functions as an accessory protein to a V-ATP-ase responsible for acidic pH maintenance in intracellular compartments, and recent data indicate that PRR could be a regulator of Wnt signaling. A circulating fragment (sPRR) is generated by enzymatic cleavage of the extracellular domain and can be quantified in plasma, serum and urine. Objectives. This is a pilot study, to explore the serum concentrations of the soluble (pro)renin receptor (sPRR) in patients with systemic sclerosis (SSc). Patients and methods. Serum samples from 29 subjects with a confirmed diagnosis of SSc have been tested using an ELISA method. Clinical and laboratory parameters have been analysed for associations with sPRR serum levels. Results and conclusion. Serum levels of sPRR were higher in patients with a history of digital ulcers (p 0.041, Mann-Whitney U-test) and those with digital pitting scars (p 0.037, Mann-Whitney U-test). Serum levels of sPRR correlated with serum creatinine (r 0.424, Spearman test), in line with previously reported data. Our results indicate the need for a larger study to assess the sPRR changes in patients with SSc, including more cases with severe disease and excluding a possible impact of angiotensin-receptor blockers.
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Krummenacher, Claude, Isabelle Baribaud, Manuel Ponce de Leon, J. Charles Whitbeck, Huan Lou, Gary H. Cohen i Roselyn J. Eisenberg. "Localization of a Binding Site for Herpes Simplex Virus Glycoprotein D on Herpesvirus Entry Mediator C by Using Antireceptor Monoclonal Antibodies". Journal of Virology 74, nr 23 (1.12.2000): 10863–72. http://dx.doi.org/10.1128/jvi.74.23.10863-10872.2000.
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ABSTRACT The human herpesvirus entry mediator C (HveC), also known as the poliovirus receptor-related protein 1 (PRR1) and as nectin-1, allows the entry of herpes simplex virus type 1 (HSV-1) and HSV-2 into mammalian cells. The interaction of virus envelope glycoprotein D (gD) with such a receptor is an essential step in the process leading to membrane fusion. HveC is a member of the immunoglobulin (Ig) superfamily and contains three Ig-like domains in its extracellular portion. The gD binding site is located within the first Ig-like domain (V domain) of HveC. We generated a panel of monoclonal antibodies (MAbs) against the ectodomain of HveC. Eleven of these, which detect linear or conformational epitopes within the V domain, were used to map a gD binding site. They allowed the detection of HveC by enzyme-linked immunosorbent assay, Western blotting, and biosensor analysis or directly on the surface of HeLa cells and human neuroblastoma cell lines, as well as simian Vero cells. The anti-HveC V-domain MAbs CK6, CK8, and CK41, as well as the previously described MAb R1.302, blocked HSV entry. Their binding to soluble HveC was blocked by the association of gD with the receptor, indicating that their epitopes overlap a gD binding site. Competition assays on an optical biosensor showed that CK6 and CK8 (linear epitopes) inhibited the binding of CK41 and R1.302 (conformational epitopes) to HveC and vice versa. Epitope mapping showed that CK6 and CK8 bound between residues 80 and 104 of HveC, suggesting that part of the gD binding site colocalizes in the same region.
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Martinez, Wanda M., i Patricia G. Spear. "Structural Features of Nectin-2 (HveB) Required for Herpes Simplex Virus Entry". Journal of Virology 75, nr 22 (15.11.2001): 11185–95. http://dx.doi.org/10.1128/jvi.75.22.11185-11195.2001.
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ABSTRACT One step in the process of herpes simplex virus (HSV) entry into cells is the binding of viral glycoprotein D (gD) to a cellular receptor. Human nectin-2 (also known as HveB and Prr2), a member of the immunoglobulin (Ig) superfamily, serves as a gD receptor for the entry of HSV-2, variant forms of HSV-1 that have amino acid substitutions at position 25 or 27 of gD (for example, HSV-1/Rid), and porcine pseudorabies virus (PRV). The gD binding region of nectin-2 is believed to be localized to the N-terminal variable-like (V) Ig domain. In order to identify specific amino acid sequences in nectin-2 that are important for HSV entry activity, chimeric molecules were constructed by exchange of sequences between human nectin-2 and its mouse homolog, mouse nectin-2, which mediates entry of PRV but not HSV-1 or HSV-2. The nectin-2 chimeric molecules were expressed in Chinese hamster ovary cells, which normally lack a gD receptor, and tested for cell surface expression and viral entry activity. As expected, chimeric molecules containing the V domain of human nectin-2 exhibited HSV entry activity. Replacement of either of two small regions in the V domain of mouse nectin-2 with amino acids from the equivalent positions in human nectin-2 (amino acids 75 to 81 or 89) transferred HSV-1/Rid entry activity to mouse nectin-2. The resulting chimeras also exhibited enhanced HSV-2 entry activity and gained the ability to mediate wild-type HSV-1 entry. Replacement of amino acid 89 of human nectin-2 with the corresponding mouse amino acid (M89F) eliminated HSV entry activity. These results identify two different amino acid sequences, predicted to lie adjacent to the C′ and C" beta-strands of the V domain, that are critical for HSV entry activity. This region is homologous to the human immunodeficiency virus binding region of CD4 and to the poliovirus binding region of CD155.
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HOU, Jie, Rui LI, Hongfang MA, Songlin QIAO i Gaiping ZHANG. "Structural prediction of porcine sialoadhesin V-set Ig-like domain sheds some light on its role in porcine reproductive and respiratory syndrome virus (PRRSV) infection". Frontiers of Agricultural Science and Engineering 3, nr 1 (2016): 65. http://dx.doi.org/10.15302/j-fase-2016086.
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Haynes, Andrew, Andrew McMillan, Andrew Jack, Wendi Qian, Ira Jakupovic, Paul Smith i Archie Prentice. "A Prospective, Randomised Trial of Chlorambucil, Mitoxantrone and Dexamethasone (CMD) Versus Fludarabine, Mitoxantrone and Dexamethasone (FMD) for Advanced Follicular Lymphoma (Real Grades I-III, Stages III/IV)." Blood 108, nr 11 (16.11.2006): 534. http://dx.doi.org/10.1182/blood.v108.11.534.534.
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Abstract Fludarabine, alone and in combination, has activity in de novo and relapsed/refractory follicle centre cell lymphoma (FCCL), as do many other combinations of chemotherapy, including chlorambucil, mitoxantrone and steroids. In this multicentre study, between May 2000 and April 2006, 400 adults with FCCL (REAL grades I-III) were randomised to receive either CMD (chlorambucil 10mgs po, od, days 1–10, mitoxantrone 12mgs/m2 IV, day 1 and dexamethasone 20mgs po, od, days 1–5) or FMD (fludarabine 25mgs/m2, IV days 1–3, with M and D as for CMD) on physicians’ decision to treat advanced stage (III or IV) disease (B symptoms, bone marrow failure, bulk or progression and compression syndromes). Rituximab (R) was not available for this trial. Randomisation was prospectively stratified by IPI score. Patients received four courses before assessment of response, then had no further trial therapy with progression or inadequate response or had a maximum total of eight courses if responding. The primary end points were progression-free survival (PFS) and overall survival (OS) and the secondary end points were complete (C) and partial (P) remission rates (RR). This preliminary report is at median follow-up of 31 months (range 0–71). Clinical features and pathological diagnoses (reviewed centrally) were equivalent across both arms. The median age in years (range; number >60 y) for CMD was 56 (32–72; 68) and for FMD 55 (29–75; 68) and 70% in both arms had stage IV FCCL. For CMD versus FMD, the CRR and PRR after course 4 were 18% v 21% and 72% v 68% and after course 8 or at the end of treatment were 44% v 45% and 47% v 47%. The hazard ratios were in favour of CMD, for OS at 1.65 (95%CI 0.97–2.8, p=0.063) and for PFS at 1.61 (95%CI 1.2–2.17; p=0.0018). The numbers of deaths due to lymphoma were equivalent (19 v 21) but there were fewer deaths in the first year (most due to lymphoma in both arms) with CMD (7 v 15; ns). The difference between those who received between 6 and 8 courses in either arm was not significant (152[80%] v 135[71%]). With CMD there were higher rates of grade 3/4 neutropenia (8 v 1) and infections (29 v 20) but these differences were not statistically significant; other toxicities were low grade and equivalent for both arms. Days between cycles (median 28 days in both arms) and dose adherence (96% and 97%) were equivalent; both regimens were well tolerated and relatively easy to deliver. The equivalent dose intensity, toxicities and RRs of the two arms indicate that the statistically significant superior PFS for CMD was not due to undertreatment in the FMD arm. The outcomes for CMD are at least as good as those reported for other combinations of chemotherapy without R and R-CVP.
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Amit, Mr. "Performance Study of Air Driven Engine Being Modified From Conventional Four Stroke Engine without Cam Modification". International Journal of Engineering and Advanced Technology 10, nr 2 (30.12.2020): 250–54. http://dx.doi.org/10.35940/ijeat.b2076.1210220.
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This research presents a novel mechanism to convert a conventional100-cc four stroke driven engine into a 2 stroke compressed air driven engine without doing any modifications in the cam shaft. This allows for a faster valve operation & also helping the existing engines to be converted into air driven engines with removal of intake and exhaust manifold and also provides a platform to curb the growing menace of air pollution by using the existing old engines to be used as air driven engine. Also, cycle of operation of the Air Engine has been representedon P-V diagram and theoretical efficiency has been calculated which is a function of pressure ratio and temperature, and comes out to be around 80 percent (@ Prr. Ratio of 10& Temp of 550K). Variation of rpm with percentage valve opening is seen at different pressure ratios and the maximum speed of the engine (after doing said modifications and running through compressed air)is observed to be 1573 rpm at a pressure of 9 bar with the developed mechanism for the existing 4 stroke maestro engine.