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1
Garcia, Natalia, Ayman Al-Hendy, Edmund C. Baracat, Katia Candido Carvalho, and Qiwei Yang. "Targeting Hedgehog Pathway and DNA Methyltransferases in Uterine Leiomyosarcoma Cells." Cells 10, no. 1 (December 31, 2020): 53. http://dx.doi.org/10.3390/cells10010053.
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Uterine leiomyosarcoma (LMS) is an aggressive tumor that presents a poor prognosis, high rates of recurrence, and metastasis. Because of its rarity, there is no information available concerning LMS molecular mechanisms of origin and development. Here, we assessed the expression profile of Hedgehog (HH) signaling pathway markers and the effects of their pharmacological inhibition on uterine smooth muscle (UTSM), leiomyoma, and LMS cells. Additionally, we also evaluated the effects of DNMTs inhibition on LMS cell behavior. Cell proliferation, migration and apoptosis rates were evaluated by MTT, Scratch, and Annexin V assays, respectively. RNA expression and protein levels were assessed by qRT-PCR and Western blot. We found that SMO and GLIs (1, 2, and 3) expression was upregulated in LMS cells, with increased nuclear levels of GLI proteins. Treatment with LDE225 (SMOi) and Gant61 (GLIi) resulted in a significant reduction in Glis protein levels in LMS (p < 0.05). Additionally, the expression of DNMT (1, 3a, and 3b), as well as GLI1 nuclear expression, was significantly decreased after treatment with HH inhibitor in LMS cells. Our results showed that blocking of SMO, GLI, and DNMTs is able to inhibit LMS proliferation, migration, and invasion. Importantly, the combination of those treatments exhibited a potentiated effect on LMS malignant features due to HH pathway deactivation.
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Plaat, Boudewijn E. C., Harry Hollema, Willemina M. Molenaar, Gerben H. Torn Broers, Justin Pijpe, Mirjam F. Mastik, Harald J. Hoekstra, Eva van den Berg, Rik J. Scheper, and Winette T. A. van der Graaf. "Soft Tissue Leiomyosarcomas and Malignant Gastrointestinal Stromal Tumors: Differences in Clinical Outcome and Expression of Multidrug Resistance Proteins." Journal of Clinical Oncology 18, no. 18 (September 18, 2000): 3211–20. http://dx.doi.org/10.1200/jco.2000.18.18.3211.
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PURPOSE: Several studies have reported clinical behavior and chemotherapy resistance in leiomyosarcomas, but these studies did not differentiate between soft tissue leiomyosarcomas (LMS) and malignant gastrointestinal stromal tumors (GIST). Multidrug resistance (MDR) has been associated with the expression of P-glycoprotein (P-gp), multidrug resistance protein (MRP1), and lung resistance protein (LRP). The aim of the present study was to compare LMS and GIST with respect to clinical outcome and MDR parameters. PATIENTS AND METHODS: Clinical outcome was evaluated in 29 patients with a primary deep-seated LMS and 26 patients with a primary malignant GIST. Paraffin-embedded material, available for 26 patients with LMS and 25 with GIST, was used for immunohistochemical detection of P-gp, MRP1, LRP, and c-kit. RESULTS: Mean overall survival (OS) was 72 months for LMS patients and 31 months for GIST patients (P < .05). Metastases occurred in 16 (59%) of 27 assessable LMS patients and in 10 (56%) of 18 assessable GIST patients. LMS predominantly metastasized to the lungs (14 of 16 patients), whereas GIST tended to spread to the liver (five of 10 patients) and the abdominal cavity (three of 10 patients; P < .001). P-gp and MRP1 expression was more pronounced in GIST than in LMS (P < .05): the mean percentage of P-gp expressing cells was 13.4% in patients with LMS and 38.4% in patients with GIST, and the mean percentage MRP1 expressing cells was 13.3% in patients with LMS and 35.4% in patients with GIST. LRP expression did not differ between LMS and GIST. c-kit was expressed in 5% of the LMS patients and in 68% of the GIST patients. CONCLUSION: LMS patients have a better survival than GIST patients, and the metastatic pattern is different. Expression of MDR proteins in LMS is less pronounced than in GIST.
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Sparić, Radmila, Mladen Andjić, Ivana Babović, Lazar Nejković, Milena Mitrović, Jelena Štulić, Miljan Pupovac, and Andrea Tinelli. "Molecular Insights in Uterine Leiomyosarcoma: A Systematic Review." International Journal of Molecular Sciences 23, no. 17 (August 27, 2022): 9728. http://dx.doi.org/10.3390/ijms23179728.
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Uterine fibroids (UFs) are the most common benign tumors of female genital diseases, unlike uterine leiomyosarcoma (LMS), a rare and aggressive uterine cancer. This narrative review aims to discuss the biology and diagnosis of LMS and, at the same time, their differential diagnosis, in order to distinguish the biological and molecular origins. The authors performed a Medline and PubMed search for the years 1990–2022 using a combination of keywords on the topics to highlight the many genes and proteins involved in the pathogenesis of LMS. The mutation of these genes, in addition to the altered expression and functions of their enzymes, are potentially biomarkers of uterine LMS. Thus, the use of this molecular and protein information could favor differential diagnosis and personalized therapy based on the molecular characteristics of LMS tissue, leading to timely diagnoses and potential better outcomes for patients.
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Ceausu, Amalia Raluca, Alexandru Ciolofan, Alexandru Blidisel, Andrei Alexandru Cosma, Pusa Nela Gaje, and Octavian Cretu. "Chloride Intracellular Channel Protein 1 Expression and Angiogenic Profile of Liver Metastasis of Digestive Origin." Current Issues in Molecular Biology 45, no. 2 (February 6, 2023): 1396–406. http://dx.doi.org/10.3390/cimb45020091.
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Chloride intracellular channel 1 (CLIC1) is involved in cell migration and metastasis. The histological growth patterns of liver metastasis are as follows: desmoplastic (d-HGP), replacement (r-HGP), pushing (p-HGP), and mixed. The aim of this study was to evaluate the relation between HGP, angiogenesis, and CLIC1 expression. Materials and Methods: A total of 40 cases of primary tumors and their LM: d-HGP (12 cases), r-HGP (13 cases), and p-HGP (15 cases), were evaluated through simple and double immunostaining. CLIC1 assessment was conducted as follows: scores of 0 (less than 10% of positive cells), 1 (10–30%), 2 (30–50%), or 3 (more than 50%) were assigned. Heterogeneous CLIC1 expression was found. CLIC1 in primary tumors correlated with grade G for all cases of LM with a p-HGP (p = 0.004). The CLIC1 score for LMs with an r-HGP correlated with grade G of the corresponding primary tumor (p = 0.027). CLIC1 and CD34+/Ki67+ vessels (p = 0.006) correlated in primary tumors. CLIC1 in primary tumors correlated with CD34+/Ki67+ vessels of LMs with a d HGP (p = 0.024). Conclusions: The CLIC1 score may have prognostic value, mainly for LMs with a p-HGP and r-HGP, and therapeutic value for LMs with a d-HGP.
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Safonova, L. A., M. M. Bobrova, O. I. Agapova, A. Yu Arkhipova, A. V. Goncharenko, and I. I. Agapov. "FIBROIN SILK BASED FILMS FOR RAT’S FULL-THICKNESS SKIN WOUND REGENERATION." Russian Journal of Transplantology and Artificial Organs 18, no. 3 (November 17, 2016): 74–84. http://dx.doi.org/10.15825/1995-1191-2016-3-74-84.
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Aimof this study is to research an effect of silk fi broin fi lms fabricated by casting method upon Wistar rat’s full-thickness skin wound regeneration.Materials and methods.4 different kinds of fi lms with protein concentration equal to 20 mg/ml were fabricated: fi lms from silk fi broin aqueous solution, fi lms from silk fi broin formic acid solution, fi lms from silk fi broin aqueous solution containing 30% collagen by weight, fi lms from silk fi broin formic acid solution containing 30% collagen by weight. All kinds of fi lms were fabricated by casting method on polished Tefl on surface. Scanning electron microscopy was applied to research fi lms’ surface structure. Cytotoxicity test of the fi lms was realized on mouse 3T3 fi broblasts model by MTT assay. Manufactured fi lms were utilized to regenerate full-thickness skin wounds in Wistar rats.Results.It was shown that fi lms’ surface was characterized by micro- and nanorelief in the form of roughness. The proliferative activity of mouse 3T3 fi broblasts increased during 7 days of cytotoxicity test. Fabricated fi lms enlarge the regeneration rate of full-thickness Wistar rat skin wounds an average of 25%. Histological analysis indicated structural skin restoration without any infl ammatory tissue.Conclusion.All fabricated fi lms are non-cytotoxic and characterized by appropriate structure for the adhesion and proliferation of fi broblasts. The application of fi lms for full-thickness skin wound regeneration increases its restoration rate which is confi rmed by histological examination.
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Azzimato, Valerio, Jennifer Jager, Ping Chen, Cecilia Morgantini, Laura Levi, Emelie Barreby, André Sulen, et al. "Liver macrophages inhibit the endogenous antioxidant response in obesity-associated insulin resistance." Science Translational Medicine 12, no. 532 (February 26, 2020): eaaw9709. http://dx.doi.org/10.1126/scitranslmed.aaw9709.
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Obesity and insulin resistance are risk factors for nonalcoholic fatty liver disease (NAFLD), the most common chronic liver disease worldwide. Because no approved medication nor an accurate and noninvasive diagnosis is currently available for NAFLD, there is a clear need to better understand the link between obesity and NAFLD. Lipid accumulation during obesity is known to be associated with oxidative stress and inflammatory activation of liver macrophages (LMs). However, we show that although LMs do not become proinflammatory during obesity, they display signs of oxidative stress. In livers of both humans and mice, antioxidant nuclear factor erythroid 2–related factor 2 (NRF2) was down-regulated with obesity and insulin resistance, yielding an impaired response to lipid accumulation. At the molecular level, a microRNA-targeting NRF2 protein, miR-144, was elevated in the livers of obese insulin-resistant humans and mice, and specific silencing of miR-144 in murine and human LMs was sufficient to restore NRF2 protein expression and the antioxidant response. These results highlight the pathological role of LMs and their therapeutic potential to restore the impaired endogenous antioxidant response in obesity-associated NAFLD.
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Böhm, Michael J., Ralf Marienfeld, Daniela Jäger, Kevin Mellert, Adrian von Witzleben, Silke Brüderlein, Mathias Wittau, et al. "Analysis of the CDK4/6 Cell Cycle Pathway in Leiomyosarcomas as a Potential Target for Inhibition by Palbociclib." Sarcoma 2019 (January 21, 2019): 1–10. http://dx.doi.org/10.1155/2019/3914232.
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Leiomyosarcoma (LMS) is characterized by high genomic complexity, and to date, no specific targeted therapy is available. In a genome-wide approach, we profiled genomic aberrations in a small cohort of eight primary tumours, two relapses, and eight metastases across nine different patients. We identified CDK4 amplification as a recurrent alteration in 5 out of 18 samples (27.8%). It has been previously shown that the LMS cell line SK-LMS-1 has a defect in the p16 pathway and that this cell line can be inhibited by the CDK4 and CDK6 inhibitor palbociclib. For SK-LMS-1 we confirm and for SK-UT-1 we show that both LMS cell lines express CDK4 and that, in addition, strong CDK6 expression is seen in SK-LMS-1, whereas Rb was expressed in SK-LMS-1 but not in SK-UT-1. We confirm that inhibition of SK-LMS-1 with palbociclib led to a strong decrease in protein levels of Phospho-Rb (Ser780), a decreased cell proliferation, and G0/G1-phase arrest with decreased S/G2fractions. SK-UT-1 did not respond to palbociclib inhibition. To compare thesein vitrofindings with patient tissue samples, a p16, CDK4, CDK6, and p-Rb immunohistochemical staining assay of a large LMS cohort (n=99patients with 159 samples) was performed assigning a potential responder phenotype to each patient, which we identified in 29 out of 99 (29.3%) patients. Taken together, these data show that CDK4/6 inhibitors may offer a new option for targeted therapy in a subset of LMS patients.
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De Gregorio, Amelie, Inga Bekes, Nikolaus de Gregorio, Sophia Andres, Diego Hoffmeister, Tatjana Swerev, Thomas W. P. Friedl, Wolfgang Janni, and Florian Ebner. "Comparison of preoperative serum VEGF in leiomyosarcoma and uterus myomatosus patients: A proof of concept study." Journal of Clinical Oncology 37, no. 15_suppl (May 20, 2019): e17117-e17117. http://dx.doi.org/10.1200/jco.2019.37.15_suppl.e17117.
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e17117 Background: The preoperative differentiation of a uterine fibroid from a sarcoma is still a pending clinical problem. Currently with the suspicion of a sarcoma (LMS) the more invasive open surgical approach is recommended to minimise the risk of LMS fragmentation and distribution. In a minority of cases, a malignancy is indicated by clinical suspicion, pre-surgical imaging or routine serum blood samples (SBS). Our hypothesis postulates higher VEGF levels in LMS compared to fibroid patients in the pre-surgical SBS. To assess this hypothesis, SBS were taken from patients with the clinical suspicion of LMS after informed consent and analysed after histology confirmed the diagnosis. Methods: Case series of patients with suspected LMS over a 4year time period. Analysis was performed via SBS collected from LMS- and fibroid patients before surgery. Serum VEGF protein was measured by ELISA. The final tumor histology was obtained from the report of the institutional pathologist. VEGF-serum levels were then compared between fibroid and LMS patients using the non-parametric Mann-Whitney U test. Results: 25 patient SBS were collected prior to surgery. In 9 cases the histopathology confirmed a LMS, with heterogeneous pTNM classifications. On average, VEGF serum levels were higher in the LMS patients as compared to the fibroid patients (628,96 pg/ml vs 351,91 pg/ml; further statistics see Table 1); however, the difference was not statistically significant (Mann-Whitney-U Test, p = 0.141). Conclusions: This proof-of-concept study with a small sample size of pre-surgical SBS indicates that VEGF serum levels may be increased in patients with histologically confirmed LMS; however, larger sample sizes are needed to validate our findings. If additional studies confirm a pronounced increase in pre-surgical serum VEGF levels in LMS patients, serum VEGF levels might routinely be used to assess the risk for a LMS in patients presenting with clinical uterine fibroids. [Table: see text]
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Terés, Raul, Paula Cerdà, Ana Sebio, Pablo Gallardo, Aida Bujosa, Maria Borrell, Berta Martin, et al. "Evaluation of dystrophin expression by immunohistochemistry as a prognostic factor in leiomyosarcomas (LMS)." Journal of Clinical Oncology 39, no. 15_suppl (May 20, 2021): e23525-e23525. http://dx.doi.org/10.1200/jco.2021.39.15_suppl.e23525.
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e23525 Background: Leiomyosarcomas (LMS) are soft tissue sarcomas that derive from smooth muscle cells and can arise anywhere in the body (most frequently in extremities, uterus or retroperitoneum). The clinical prognostic factors are well established but molecular factors influencing prognostic are unknown. The DMD gene, which codifies for the dystrophin protein, has been proposed as tumour suppressor gene in LMS. The aim of our study is to evaluate dystrophin expression in LMS and its relation with the patient prognosis. Methods: A total of 103 patients (pts) with LMS were analysed. Clinical and pathological data were collected retrospectively from patients’ electronic board system. Dystrophin expression was analysed by immunohistochemistry (IHC) in paraffin embedded tissue LMS samples using the Novocastra NCL-DYS2 (Leica Biosystems). Semiquantitative assessment of the staining was classified from score 0 to 3 (0 = no dystrophin expression, 1 = low expression, 2 = moderate expression, 3 = high expression). Results: Of all 103 pts, 70 (68%) had localized disease and the majority of tumours were larger than 5 cm (75%). The most frequent locations of primary LMS were extremities (n = 31; 30.1%), uterus (n = 23; 23.2%) and retroperitoneum (n = 21; 20.4%). Most LMS were high grade (G): 7 pts G1 (6.8%), 35 pts G2 (34%) 2 and 53 pts G3 (51.5%). 50 of all grade LMS (48.6%) had loss of dystrophin expression (score 0), 17 (16.5%) had score 1, 23 (22.3%) score 2 and 10 (9.7%) score 3. Loss/low dystrophin expression measured as score 0 or 1, was more frequent in grade 3 LMS compared to grade 2 or grade 1 (77.4% vs. 57.1% vs. 16.7%; p = 0.005). There were no differences in dystrophin loss between localized or metastatic disease at diagnosis (64.2% vs. 72.2%; p > 0.05). In our series, loss or reduced dystrophin expression did not correlate with the risk of relapse in localized patients or overall survival in metastatic patients. Conclusions: Loss of dystrophin expression is a common event in LMS. Loss/low dystrophin expression is more frequent in grade 3 LMS compared to grade 2 and grade 1 LMS. Loss of dystrophin expression did not correlate with risk of relapse or overall survival in our series. Additional genetic evaluations to study DMD in LMS are underway.
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Gonzalez-Molina, Jordi, Paula Hahn, Raul Maia Falcão, Georgia Kokaraki, Jorge Estefano de Souza, Tirzah Braz Petta Lajus, Kaisa Lehti, and Joseph W. Carlson. "Abstract 6096: The microarchitecture and fibrillar collagen expression of leiomyosarcoma are associated with malignancy." Cancer Research 82, no. 12_Supplement (June 15, 2022): 6096. http://dx.doi.org/10.1158/1538-7445.am2022-6096.
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Abstract The expression, abundance, and microarchitecture of fibrillar collagens are associated with tumor development and aggressiveness in various carcinomas. However, the impact of fibrillar collagens on mesenchymal tumors is less understood. While uterine leiomyomas, also known as fibroids, are characterized by high fibrillar collagen deposition and depend on ECM signaling for cell proliferation, the impact of fibrillar collagens on malignant uterine leiomyosarcomas has not been explored. Thus, identifying malignancy and aggressiveness-associated features of the fibrillar collagen-leiomyosarcoma crosstalk may provide novel biomarkers and therapeutic targets for these aggressive tumors. We used publicly available RNAseq data and performed RNAseq and picrosirius red analysis of fiber microarchitecture in a cohort of normal myometrium (MM; n =68 ), leiomyoma (LM; n = 66), and leiomyosarcoma (LMS; n = 67) tissues. Furthermore, we cultured patient-derived primary cells (4 MM, 3 LM, and 4 LMS) on collagen I-functionalized polyacrylamide gels at stiffness ranging from 0.5 to 115 kPa, covering the physiological and pathological stiffness, to investigate distinct behaviors between cell types, including proliferation, migration, and activity of the ECM stiffness molecular rheostat YAP/TAZ. At the protein level, analysis of fibrillar collagen microarchitecture revealed that LMS tumors present reduced fibrillar collagen density and hyphal growth units and enhanced fiber endpoints compared to both MM and LM. At the gene expression level, however, LMS tumors did not show reduced fibrillar collagen expression, instead they exhibited enhanced matrix metalloproteinase expression, particularly of MMP14. Furthermore, COL11A1 was specifically upregulated in LMS tumors and its expression was associated with poor prognosis. Finally, in vitro response of MM, LM, and LMS cells to collagen I at defined stiffness showed that LMS cell migration, proliferation, and subcellular localization of YAP/TAZ are less sensitive to substrate stiffness than in MM and LM cells, although the response varied between distinct donors. In conclusion, we show that LMS tumors typically present low fibrillar collagen protein expression likely due to enhanced degradation. In addition, collagen I adhesion and stiffness have a lower impact on malignant LMS cells than on MM and LM, which may explain their ability to grow in low-collagen microenvironments. Furthermore, this study shows that COL11A1 is a potential biomarker with prognostic value in leiomyosarcoma. Citation Format: Jordi Gonzalez-Molina, Paula Hahn, Raul Maia Falcão, Georgia Kokaraki, Jorge Estefano de Souza, Tirzah Braz Petta Lajus, Kaisa Lehti, Joseph W. Carlson. The microarchitecture and fibrillar collagen expression of leiomyosarcoma are associated with malignancy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 6096.
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Grassin-Delyle, Stanislas, Charlotte Abrial, Hélène Salvator, Marion Brollo, Emmanuel Naline, and Philippe Devillier. "The Role of Toll-Like Receptors in the Production of Cytokines by Human Lung Macrophages." Journal of Innate Immunity 12, no. 1 (December 17, 2018): 63–73. http://dx.doi.org/10.1159/000494463.
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Background: The Toll-like receptor (TLR) family is involved in the recognition of and response to microbial infections. These receptors are expressed in leukocytes. TLR stimulation induces the production of proinflammatory cytokines and chemokines. Given that human lung macrophages (LMs) constitute the first line of defense against inhaled pathogens, the objective of this study was to investigate the expression and function of TLR subtypes in this cell population. Methods: Human primary LMs were obtained from patients undergoing surgical resection. The RNA and protein expression levels of TLRs, chemokines, and cytokines were assessed after incubation with subtype-selective agonists. Results: In human LMs, the TLR expression level varied from one subtype to another. Stimulation with subtype-selective agonists induced an intense, concentration- and time-dependent increase in the production of chemokines and cytokines. TLR4 stimulation induced the strongest effect, whereas TLR9 stimulation induced a much weaker response. Conclusions: The stimulation of TLRs in human LMs induces intense cytokine and chemokine production, a characteristic of the proinflammatory M1 macrophage phenotype.
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Putluri, Srinivasareddy, and Md Zia Ur Rahman. "Efficient Adaptive Exon Prediction for DNA study using Proportionate LMS Variants." International Journal of Engineering & Technology 7, no. 2.17 (April 15, 2018): 116. http://dx.doi.org/10.14419/ijet.v7i2.17.11721.
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In the field of Bio-informatics, locating the exon fragments in a deoxyribonucleic acid (DNA) sequence is an important and vital work. Study of protein coding regions is a wide phenomenon in identification of diseases and design of drugs. The regions of DNA that have the protein coding information are termed as exons. Hence identifying the exon segments in a genomic sequence is a crucial job in bio-informatics. Three base periodicity (TBP) has been observed in the regions of DNA sequences can be easily determined by applying signal processing methods. Adaptive signal processing techniques found to be useful than other available methods. This is due to their unique capability to alter weight coefficients based on genomic sequence. We propose efficient adaptive exon predictors (AEPs) based on these considerations using Proportionate Normalized LMS (PNLMS) algorithm and Maximum Proportionate Normalized LMS (MPNLMS) algorithm to improve exon locating ability and better convergence. To ease the complexity of computations in the denominator during filtering process, proposed AEPs using PNLMS and its maximum variants are combined with signature algorithms. Hybrid variants of proposed AEPs include PNLMS, DCPNLMS, ECPNLMS, SSPNLMS, MPNLMS, MDCPNLMS, MECPNLMS and MSSPNLMS algorithms. It was shown that the AEP based on MDCPNLMS is superior in applications of exon identification depending on performance measures with Sensitivity 0.7346, Specificity 0.7483 and precision 0.7325 for a genomic sequence with accession AF009962 at a threshold of 0.8. Finally the capability of several AEPs in predicting exon locations is verified using different DNA sequences found in National Center for Biotechnology Information (NCBI) gene database.
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Boichuk, S. V., B. R. Ramazanov, A. R. Galembikova, O. R. Galeev, I. G. Mustafin, and A. Duensing. "DNA repair profile in gastrointestinal stromal tumors (gists) - novel perspectives for therapy." Kazan medical journal 95, no. 6 (December 15, 2014): 888–91. http://dx.doi.org/10.17816/kmj1999.
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Aim. To assess the expression of various types of DNA repair proteins in gastrointestinal stromal tumors (GISTs) to identify the possible defects in DNA repair pathways and therapeutic targets. Methods. The study was performed on the human fibroblasts, imatinib-sensitive vs imatinib-resistant GISTs and leiomyosarcomas (LMS) cell lines, as well. The cell lines indicated above were cultured in the corresponding culture medium supplemented with fetal bovine serum, L-glutamine and antibiotics (37 °C и 5% СО2). Protein expression level and its intracellular localization were assessed by Western blotting. Results. The reduced BRCA1 expression was observed in most of the GIST cell lines, which was associated with an up-regulation of Rad51, thereby indicating about the potential abnormalities of homologous recombination pathway in these cells. This phenomenon was typical for GISTs and was not observed in LMS cells lines. In contrast to LMS cell lines, all GIST cells showed an upregulation of O6-methylguanine DNA methyltransferase (MGMT), the key enzyme involved in alkylated DNA damage repair pathway. Most GIST cells exhibited high level of MSH6 known as a key member of mismatch repair pathway. Most notably, topoisomerases were over-expressed in all of GIST cell lines. Conclusions. We found several striking alterations in expression levels of DDR pathway enzymes in GISTs. For instance, an up-regulation of topoisomerases in all GISTs indicates that these cells might be sensitive to topoisomerase II inhibitors and could be potentially targeted therapeutically.
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Massimino, Luca, Alessandra Bulbarelli, Paola Antonia Corsetto, Chiara Milani, Laura Botto, Francesca Farina, Luigi Antonio Lamparelli, et al. "LSEA Evaluation of Lipid Mediators of Inflammation in Lung and Cortex of Mice Exposed to Diesel Air Pollution." Biomedicines 10, no. 3 (March 19, 2022): 712. http://dx.doi.org/10.3390/biomedicines10030712.
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Airborne ultrafine particle (UFP) exposure is a great concern as they have been correlated to increased cardiovascular mortality, neurodegenerative diseases and morbidity in occupational and environmental settings. The ultrafine components of diesel exhaust particles (DEPs) represent about 25% of the emission mass; these particles have a great surface area and consequently high capacity to adsorb toxic molecules, then transported throughout the body. Previous in-vivo studies indicated that DEP exposure increases pro- and antioxidant protein levels and activates inflammatory response both in respiratory and cardiovascular systems. In cells, DEPs can cause additional reactive oxygen species (ROS) production, which attacks surrounding molecules, such as lipids. The cell membrane provides lipid mediators (LMs) that modulate cell-cell communication, inflammation, and resolution processes, suggesting the importance of understanding lipid modifications induced by DEPs. In this study, with a lipidomic approach, we evaluated in the mouse lung and cortex how DEP acute and subacute treatments impact polyunsaturated fatty acid-derived LMs. To analyze the data, we designed an ad hoc bioinformatic pipeline to evaluate the functional enrichment of lipid sets belonging to the specific biological processes (Lipid Set Enrichment Analysis-LSEA). Moreover, the data obtained correlate tissue LMs and proteins associated with inflammatory process (COX-2, MPO), oxidative stress (HO-1, iNOS, and Hsp70), involved in the activation of many xenobiotics as well as PAH metabolism (Cyp1B1), suggesting a crucial role of lipids in the process of DEP-induced tissue damage.
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Song, Jing, Yaqi Wang, Xiao Yuan, Qiuxia Ji, Cunhui Fan, Hongmei Zhao, Wenjing Hao, and Dapeng Ren. "Stretching magnitude–dependent inactivation of AKT by ROS led to enhanced p53 mitochondrial translocation and myoblast apoptosis." Molecular Biology of the Cell 30, no. 10 (May 2019): 1182–97. http://dx.doi.org/10.1091/mbc.e18-12-0770.
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Previously, we had shown that high magnitude stretch (HMS), rather than low magnitude stretch (LMS), induced significant apoptosis of skeletal muscle C2C12 myoblasts. However, the molecular mechanism remains obscure. In this study, we found that p53 protein accumulated in the nucleus of LMS-loaded cells, whereas it translocated into mitochondria of HMS-loaded cells. Knocking down endogenous p53 by shRNA abrogated HMS-induced apoptosis. Furthermore, we demonstrated that overaccumulation of reactive oxygen species (ROS) during HMS-inactivated AKT that was activated in LMS-treated cells, which accounted for the distinct p53 subcellular localizations under HMS and LMS. Blocking ROS generation by N-acetylcysteine (NAC) or overexpressing constitutively active AKT vector (CA-AKT) inhibited HMS-incurred p53 mitochondrial translocation and promoted its nuclear targeting. Moreover, both NAC and CA-AKT significantly attenuated HMS-induced C2C12 apoptosis. Finally, we found that Ser389 phosphorylation of p53 was a downstream event of ROS-inactivated AKT pathway, which was critical to p53 mitochondrial trafficking during HMS stimuli. Transfecting p53-shRNA C2C12s with the mutant p53 (S389A) that was unable to target p53 to mitochondria underwent significantly lower apoptosis than transfection with wild-type p53. Altogether, our study uncovered that mitochondrial localization of p53, resulting from p53 Ser389 phosphorylation through ROS-inactivated AKT pathway, prompted C2C12 myoblast apoptosis during HMS stimulation.
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Walters, Matthew P., Ellen D. McPhail, Mark E. Law, and Andrew L. Folpe. "BCL-6 expression in mesenchymal tumours: an immunohistochemical and fluorescence in situ hybridisation study." Journal of Clinical Pathology 64, no. 10 (July 1, 2011): 866–69. http://dx.doi.org/10.1136/jclinpath-2011-200185.
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The BCL-6 proto-oncogene encodes a transcriptional repressor protein. Among normal tissues, BCL-6 expression is confined to germinal center B-cells and a subpopulation of T-helper cells. Little is known about BCL-6 expression in mesenchymal tissues. We examined a series of solitary fibrous tumor (SFT) and other mesenchymal tumors for BCL-6 expression. Immunohistochemistry for BCL-6 was performed on 64 mesenchymal tumors [26 SFT (19 benign/uncertain, 7 malignant), 6 synovial sarcomas (SS), 5 gastrointestinal stromal tumors (GIST), 5 malignant peripheral nerve sheath tumors (MPNST), 5 leiomyosarcomas (LMS), 9 leiomyomas (LM) 4 desmoid tumors (DT), 4 perineuriomas (PN)]. Nuclear immunoreactivity was considered positive. Six BCL-6 positive SFT were also tested for BCL-6 gene rearrangement/amplification by FISH. Nuclear expression of BCL-6 was seen in 13/26 SFT, 5/5 LMS, 1/9 LM, 5/6 SS, 1/5 GIST, 1/5 MPNST, 1/4 PN, and 0/5 DT. BCL-6 expression was significantly more frequent in malignant (6/7) as compared with benign/uncertain SFT (6/19) (p=0.02) and in LMS (5/5) as compared with LM (1/9) (p=0.003). FISH for BCL-6 rearrangement/amplification was negative in all tested cases. We have observed BCL-6 expression in 50% or more of SFT, SS, and LMS, and in a lesser percentage of LM, GIST, MPNST and PN. Significantly more frequent expression of BCL-6 in malignant compared with benign/uncertain SFT and in LMS compared with LM suggests abnormalities in the BCL-6 signaling pathway may contribute to malignant transformation in at least some mesenchymal tumors. It is unlikely that BCL-6 expression in mesenchymal tumors is due to BCL-6 gene amplification or rearrangement. amplification or rearrangement.
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Sohn, Mira, Pavlina Ivanova, H. Alex Brown, Daniel J. Toth, Peter Varnai, Yeun Ju Kim, and Tamas Balla. "Lenz-Majewski mutations in PTDSS1 affect phosphatidylinositol 4-phosphate metabolism at ER-PM and ER-Golgi junctions." Proceedings of the National Academy of Sciences 113, no. 16 (April 4, 2016): 4314–19. http://dx.doi.org/10.1073/pnas.1525719113.
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Lenz-Majewski syndrome (LMS) is a rare disease characterized by complex craniofacial, dental, cutaneous, and limb abnormalities combined with intellectual disability. Mutations in the PTDSS1 gene coding one of the phosphatidylserine (PS) synthase enzymes, PSS1, were described as causative in LMS patients. Such mutations render PSS1 insensitive to feedback inhibition by PS levels. Here we show that expression of mutant PSS1 enzymes decreased phosphatidylinositol 4-phosphate (PI4P) levels both in the Golgi and the plasma membrane (PM) by activating the Sac1 phosphatase and altered PI4P cycling at the PM. Conversely, inhibitors of PI4KA, the enzyme that makes PI4P in the PM, blocked PS synthesis and reduced PS levels by 50% in normal cells. However, mutant PSS1 enzymes alleviated the PI4P dependence of PS synthesis. Oxysterol-binding protein–related protein 8, which was recently identified as a PI4P-PS exchanger between the ER and PM, showed PI4P-dependent membrane association that was significantly decreased by expression of PSS1 mutant enzymes. Our studies reveal that PS synthesis is tightly coupled to PI4P-dependent PS transport from the ER. Consequently, PSS1 mutations not only affect cellular PS levels and distribution but also lead to a more complex imbalance in lipid homeostasis by disturbing PI4P metabolism.
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Spiekermann, Karsten, Ralf J. Dirschinger, Ruth Schwab, Ksenia Bagrintseva, Florian Faber, Christian Buske, Susanne Schnittger, Louise M. Kelly, D. Gary Gilliland, and Wolfgang Hiddemann. "The protein tyrosine kinase inhibitor SU5614 inhibits FLT3 and induces growth arrest and apoptosis in AML-derived cell lines expressing a constitutively activated FLT3." Blood 101, no. 4 (February 15, 2003): 1494–504. http://dx.doi.org/10.1182/blood-2002-04-1045.
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Activating mutations of the protein tyrosine kinase (PTK) FLT3 can be found in approximately 30% of patients with acute myeloid leukemia (AML), thereby representing the most frequent single genetic alteration in AML. These mutations occur in the juxtamembrane (FLT3 length mutations; FLT3-LMs) and the second tyrosine kinase domain of FLT3-TKD and confer interleukin 3 (IL-3)–independent growth to Ba/F3 cells. In the mouse bone marrow transplantation model, FLT3-LMs induce a myeloproliferative syndrome stressing their transforming activity in vivo. In this study, we analyzed the pro-proliferative and antiapoptotic potential of FLT3 in FLT3-LM/TKD-mutation–transformed Ba/F3 cells and AML-derived cell lines. The PTK inhibitor SU5614 has inhibitory activity for FLT3 and selectively induces growth arrest, apoptosis, and cell cycle arrest in Ba/F3 and AML cell lines expressing a constitutively activated FLT3. In addition, the compound reverts the antiapoptotic and pro-proliferative activity of FLT3 ligand (FL) in FL-dependent cells. No cytotoxic activity of SU5614 was found in leukemic cell lines that express a nonactivated FLT3 or no FLT3 protein. At the biochemical level, SU5614 down-regulated the activity of the hyperphosphorylated FLT3 receptor and its downstream targets, signal transducer and activator of (STAT) 3, STAT5, and mitogen-activated protein kinase (MAPK), and the STAT5 target genes BCL-XL and p21. Our results show that SU5614 is a PTK inhibitor of FLT3 and has antiproliferative and proapoptotic activity in AML-derived cell lines that endogenously express an activated FLT3 receptor. The selective and potent cytotoxicity of FLT3 PTK inhibitors support a clinical strategy of targeting FLT3 as a new molecular treatment option for patients with FLT3-LM/TKD-mutation+ AML.
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Lin, Andrew, Qin Zhou, Sean Devlin, Nelly G. Adel, and Dan Douer. "Outcomes Of Tyrosine Kinase Inhibitors In Combination With Various Treatments Regimens In Adult Patients With Philadelphia Chromosome Positive Acute Lymphocytic Leukemia." Blood 122, no. 21 (November 15, 2013): 2644. http://dx.doi.org/10.1182/blood.v122.21.2644.2644.
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Abstract Background Philadelphia chromosome positive (Ph+) acute lymphoblastic leukemia (ALL) is characterized by t(9;22) and is associated with poorer outcomes. Ph+ ALL responds initially to conventional chemotherapy but overall survival (OS) is low at < 20% with no curative form of post-remission chemotherapy. Allogeneic hematopoietic stem cell transplantation (HSCT) in first remission has historically been the only curative treatment modality, yet only cures approximately 30% of patients, typically in younger patients (less than 60 years of age). The introduction of tyrosine kinase inhibitors (TKIs) that target the fusion protein Bcr-ABL have altered the management of Ph+ ALL, with durable remission of 40-50%. Debate remains over the optimal chemotherapy “backbone” to combine with TKIs in the treatment of Ph+ ALL, given outcomes reported using nonmyeloablative approaches (Foà R, et al. Blood 2011; 118:6521). Methods We conducted a single-center, retrospective analysis of patients with newly diagnosed Ph+ ALL treated with various induction regimens in combination with TKIs. Patients aged 18 or older were included and comparisons were made between regimens considered to be intensely myelosuppressive (IMS) (i.e. hyperCVAD or ALL-2) and regimens considered less myelosuppressive (LMS) (i.e. ECOG 2993, L-20, or vincristine ± corticosteroids). The primary endpoint was a comparison of overall survival between treatment groups. Secondary endpoints included assessment of complete remission (CR) rates, disease free survival (DFS), likelihood to proceed to allogeneic HSCT, the effect of HSCT on overall survival, and hematologic and non-hematologic toxicities. Results Twenty-seven patients were included in this analysis. In combination with imatinib or dasatinib, eighteen patients were treated with IMS regimens while nine patients received LMS regimens. The groups were well-matched in baseline characteristics other than age (median: IMS - 49.6 years old vs. LMS -70.6 years old, p = 0.022). CR rates were high in both treatment groups (IMS - 94.4% vs. LMS -88.9%). No difference was observed in overall survival between the groups (3-year OS rate: IMS - 43.2% vs. LMS - 44.4%, Log-rank p = 0.921). There was a nonsignificant difference in DFS for patients favoring with IMS regimens (3-year DFS rate: IMS -43.2% vs. LMS - 33.3%, Log-rank p = 0.606). Patients treated with IMS regimens were more likely to proceed to transplant in CR1 [IMS - 88.2% (n = 15/17) vs. LMS - 22.2% (n = 2/ 9), p = 0.002). When evaluating the impact of HSCT on 3-year OS using a landmark analysis of 6 months post-induction therapy there was no difference between 13 patients who proceeded to HSCT compared to the 9 who did not (3-year OS rate post landmark: Yes HSCT - 33.7% vs. No HSCT - 55.6%, Log-rank p = 0.4). Overall, the toxicity during induction was similar in both treatment groups despite the increased myelosuppression that was expected in the more intense treatment group (100% experience grade 3 or greater hematologic toxicity vs. 89.9%). Among patients who achieve ANC>1000, those receiving LMS regimens did have a decrease in time to achieve this benchmark, though statistically it was not significant (median: 16 days vs. 21 days, p = 0.243). Conclusion While limited by the small sample size, it appears that the combination of TKIs with either IMS or LMS treatment regimens will achieve high CR rates. In our study OS was improved compared to historical controls prior to the introduction of TKIs and this includes a number of patients who did not undergo HSCT (Fielding AK, et al. Blood 2009; 113:4489-4496). It appears that in older patients who receive LMS regimens may not have inferior outcomes compared to younger patients who received IMS despite being far less likely to undergo HSCT in CR1. This finding may potentially prove to be significant given that up to 50% of patients with ALL older than 60 years of age are Ph+. Disclosures: No relevant conflicts of interest to declare.
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Dong, Jun, Titus J. Boggon, Naomi E. Chayen, James Raftery, Ru-Chang Bi, and John R. Helliwell. "Bound-solvent structures for microgravity-, ground control-, gel- and microbatch-grown hen egg-white lysozyme crystals at 1.8 Å resolution." Acta Crystallographica Section D Biological Crystallography 55, no. 4 (April 1, 1999): 745–52. http://dx.doi.org/10.1107/s0907444998016047.
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A number of methods can be used to improve the stability of the protein crystal-growth environment, including growth in microgravity without an air–liquid phase boundary, growth in gels and growth under oil (`microbatch'). In this study, X-ray data has been collected from and structures refined for crystals of hen egg-white lysozyme (HEWL) grown using four different methods, liquid–liquid dialysis on Earth and in microgravity using the European Space Agency's (ESA) Advanced Protein Crystallization Facility (APCF) on board the NASA Space Shuttle Life and Microgravity Spacelab (LMS) mission (STS-78), crystallization in agarose gel using a tube liquid–gel diffusion method and crystallization in microbatch under oil. A comparison of the overall quality of the X-ray data, the protein structures and especially the bound-water structures has been carried out at 1.8 Å. The lysozyme protein structures corresponding to these four different crystallization methods remain similar. A small improvement in the bound-solvent structure is seen in lysozyme crystals grown in microgravity by liquid–liquid dialysis, which has a more stable fluid physics state in microgravity, and is consistent with a better formed protein crystal in microgravity.
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Seda, Marian, Emma Peskett, Charalambos Demetriou, Dale Bryant, Gudrun E. Moore, Philip Stanier, and Dagan Jenkins. "Analysis of transgenic zebrafish expressing the Lenz-Majewski syndrome gene PTDSS1 in skeletal cell lineages." F1000Research 8 (March 11, 2019): 273. http://dx.doi.org/10.12688/f1000research.17314.1.
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Background: Lenz-Majewski syndrome (LMS) is characterized by osteosclerosis and hyperostosis of skull, vertebrae and tubular bones as well as craniofacial, dental, cutaneous, and digit abnormalities. We previously found that LMS is caused by de novo dominant missense mutations in the PTDSS1 gene, which encodes phosphatidylserine synthase 1 (PSS1), an enzyme that catalyses the conversion of phosphatidylcholine to phosphatidylserine. The mutations causing LMS result in a gain-of-function, leading to increased enzyme activity and blocking end-product inhibition of PSS1. Methods: Here, we have used transpose-mediated transgenesis to attempt to stably express wild-type and mutant forms of human PTDSS1 ubiquitously or specifically in chondrocytes, osteoblasts or osteoclasts in zebrafish. Results: We report multiple genomic integration sites for each of 8 different transgenes. While we confirmed that the ubiquitously driven transgene constructs were functional in terms of driving gene expression following transient transfection in HeLa cells, and that all lines exhibited expression of a heart-specific cistron within the transgene, we failed to detect PTDSS1 gene expression at either the RNA or protein levels in zebrafish. All wild-type and mutant transgenic lines of zebrafish exhibited mild scoliosis with variable incomplete penetrance which was never observed in non-transgenic animals. Conclusions: Collectively the data suggest that the transgenes are silenced, that animals with integrations that escape silencing are not viable, or that other technical factors prevent transgene expression. In conclusion, the incomplete penetrance of the phenotype and the lack of a matched transgenic control model precludes further meaningful investigations of these transgenic lines.
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Dechantsreiter, Susanne, Ashley Ambrose, Edith M. Hessel, Soren Beinke, Gillian Tannahill, and Daniel M. Davis. "Activation-induced cell protrusions and vesicle secretion of human lung macrophages revealed by super-resolution microscopy." Journal of Immunology 204, no. 1_Supplement (May 1, 2020): 149.1. http://dx.doi.org/10.4049/jimmunol.204.supp.149.1.
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Abstract Recent studies using super-resolution microscopy have established that the organisation of immune cell receptors impacts signal integration and cellular activation. Understanding the nano-scale dynamics of surface receptors on tissue specific macrophages is especially important, as they are phenotypically diverse and possess a large repertoire of receptors. However, tissue macrophages are highly auto-fluorescent, severely limiting the utility of light microscopy. Here, we report a novel correction technique which utilises a moving median filter to remove auto-fluorescent noise from stochastic optical reconstruction microscopy datasets. Using this, we visualised lung macrophages (LMs) activated through Fc receptors by IgG-coated glass slides, representing a 2D model of the phagocytic synapse. This unexpectedly revealed the formation of protrusions, at the surface of LMs but not blood-derived macrophages. Class I MHC protein accumulated at the tips, consistent with a role for macrophage protrusions in antigen presentation. Additionally, staining for the exosome marker CD81 revealed the secretion of extracellular vesicles. Classically, cell-derived vesicles are studied after bulk isolation, which is harsh and comes with many caveats. Imaging them directly upon secretion allows analysis of their properties on a cell-by-cell basis in a near-native state. We discovered that LM vesicles appeared distinct from those secreted from blood-derived macrophages in that their average diameter was much smaller (80 nm ± 19 nm vs. 159 nm ± 78 nm). Thus, our correction method for super-resolution microscopy revealed novel cell biology – protrusion formation and vesicle secretion – triggered upon activation of human LMs.
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Refet-Mollof, Elena, Ouafa Najyb, Rodin Chermat, Audrey Glory, Julie Lafontaine, Philip Wong, and Thomas Gervais. "Hypoxic Jumbo Spheroids On-A-Chip (HOnAChip): Insights into Treatment Efficacy." Cancers 13, no. 16 (August 11, 2021): 4046. http://dx.doi.org/10.3390/cancers13164046.
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Hypoxia is a key characteristic of the tumor microenvironment, too rarely considered during drug development due to the lack of a user-friendly method to culture naturally hypoxic 3D tumor models. In this study, we used soft lithography to engineer a microfluidic platform allowing the culture of up to 240 naturally hypoxic tumor spheroids within an 80 mm by 82.5 mm chip. These jumbo spheroids on a chip are the largest to date (>750 µm), and express gold-standard hypoxic protein CAIX at their core only, a feature absent from smaller spheroids of the same cell lines. Using histopathology, we investigated response to combined radiotherapy (RT) and hypoxic prodrug Tirapazamine (TPZ) on our jumbo spheroids produced using two sarcoma cell lines (STS117 and SK-LMS-1). Our results demonstrate that TPZ preferentially targets the hypoxic core (STS117: p = 0.0009; SK-LMS-1: p = 0.0038), but the spheroids’ hypoxic core harbored as much DNA damage 24 h after irradiation as normoxic spheroid cells. These results validate our microfluidic device and jumbo spheroids as potent fundamental and pre-clinical tools for the study of hypoxia and its effects on treatment response.
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Velebný, S., G. Hrčkova, and G. Kogan. "Impact of treatment with praziquantel, silymarin and/or β-glucan on pathophysiological markers of liver damage and fibrosis in mice infected with Mesocestoides vogae (Cestoda) tetrathyridia." Journal of Helminthology 82, no. 3 (September 2008): 211–19. http://dx.doi.org/10.1017/s0022149x08960776.
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AbstractMesocestoides vogae tetrathyridia infection in mice causes hepatocyte injury, hepatic granulomatous inflammmation, liver fibrosis and chronic peritonitis manifested with portal hypertension. To reduce the detrimental effect of parasites on the host liver, the effect of the anthelmintic drug praziquantel (PZQ) in combination with natural products silymarin (an antioxidant) and β-glucan (an immunomodulator) was investigated. The therapeutic effect of drugs was assessed by means of aminotransferase (alanine aminotransferase (ALT) and aspartate aminotransferase (AST)) activities, content of albumin, total proteins and hyaluronic acid (HA) in sera of ICR mice infected with M. vogae larvae. Animals were treated with PZQ suspended in oil emulsion (Group 1), PZQ combined with silymarin incorporated into lipid microspheres (LMS) (Group 2), PZQ combined with β-glucan incorporated into liposomes (LG) (Group 3), PZQ co-administered with LMS and LG (Group 4). Untreated animals (Group 5) served as the control. Treatment of animals started at the early chronic phase of infection (day 14 p.i.) and lasted 10 days; serum samples were collected on days 0, 7, 14, 25, 28, 31, 35 and 45 p.i. ALT and AST activities were significantly (P < 0.05) decreased in Groups 2, 3 and 4. HA content was significantly (P < 0.05 and 0.01) lower in Groups 2 and 4. Albumin levels were decreased in Groups 2 and 4, total protein concentration decreased in Groups 1 and 3 (P < 0.05 and 0.01). These results showed that combined treatment of PZQ with silymarin and/or β-glucan was able to ameliorate or suppress fibrogenesis in the liver, protect liver cells from oxidative damage and, possibly, stimulate regeneration of the parenchyma.
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Esposito, L., F. Sica, G. Sorrentino, R. Berisio, L. Carotenuto, A. Giordano, C. A. Raia, et al. "Protein Crystal Growth in the Advanced Protein Crystallization Facility on the LMS Mission: a Comparison of Sulfolobus solfataricus Alcohol Dehydrogenase Crystals Grown on the Ground and in Microgravity." Acta Crystallographica Section D Biological Crystallography 54, no. 3 (May 1, 1998): 386–90. http://dx.doi.org/10.1107/s0907444997011992.
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Crystals of alcohol dehydrogenase from Sulfolobus solfataricus were grown in the Advanced Protein Crystallization Facility during the Life and Microgravity Sciences Spacelab mission on the US Space Shuttle. Large diffracting crystals were obtained by dialysis, whereas only poor-quality crystals were obtained by vapour diffusion. The quality of both the microgravity and ground-based crystals was analysed by X-ray diffraction. There was some improvement in terms of size and diffraction resolution limit for the microgravity crystals. However, the twinning observed in the Earth-grown crystals was also present for those grown in microgravity.
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Frost, D., J. Lasota, and M. Miettinen. "Gastrointestinal Stromal Tumors and Leiomyomas in the Dog: A Histopathologic, Immunohistochemical, and Molecular Genetic Study of 50 Cases." Veterinary Pathology 40, no. 1 (January 2003): 42–54. http://dx.doi.org/10.1354/vp.40-1-42.
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Fifty canine gastrointestinal (GI) mesenchymal tumors were examined to determine the occurrence of leiomyomas (LM) and GI stromal tumors and to compare their clinicopathologic features. Twenty-one tumors (42%) were histologically reclassified as gastrointestinal stromal tumors (GISTs) and 29 tumors (58%) as LMs on the basis of their histologic similarity with homologous human tumors. The GISTs occurred equally in males and females, with a mean age of 11 years (range 5–14 years). Five GISTs (24%) were associated with clinical signs and six (29%) had metastasis in liver or abdominal cavity. The GISTs occurred in large intestine (10, 48%), small bowel (six, 29%), stomach (four, 19%), and mesentery of small intestine (one, 5%). Histologically, they were highly cellular spindle, or less commonly epithelioid tumors with mitotic rates ranging from 0 to 19 per 10 HPF. Eleven tumors (52%) were positive for CD117 (KIT); seven (33%) were positive for smooth muscle actin but none for desmin and S-100 protein. Sequences of KIT exon 11, often mutated in human GISTs, were evaluated from four GISTs. Deletion of Try556-Lys557 coexisting with duplication of Gln555 in one case of GIST and T to C transition resulting in substitution of Pro for Leu575 in another were identified. The LMs occurred predominantly in males (82%) with a mean age of 11 years (range 8–17 years). Nine tumors (31%) had associated clinical signs. They occurred in the stomach (22, 76%), esophagus (four, 14%), and intestines (three, 10%); all were paucicellular, had no mitoses, and were composed of mature smooth muscle cells. Twenty-eight (97%) were positive for smooth muscle actin and 18(62%) for desmin but none for CD117 and S-100. Both GISTs and true LMs occur in the GI tract of dogs. Both tumors have distinctive pathologic features.
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Grosso, F., R. Sanfilippo, R. L. Jones, P. Collini, C. Morosi, F. Raspagliesi, J. C. Tercero, M. D'Incalci, I. R. Judson, and P. G. Casali. "Role of trabectedin (T) in the management of advanced uterine leiomyosarcoma (U-LM)." Journal of Clinical Oncology 27, no. 15_suppl (May 20, 2009): 10530. http://dx.doi.org/10.1200/jco.2009.27.15_suppl.10530.
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10530 Background: To explore the clinical impact of T in U-LM. T has been approved in Europe for second line treatment of advanced soft tissue sarcomas (STS). Efficacy is well established in liposarcoma and leiomyosarcoma. U-LMs display peculiar clinical and genetic features compared to other STS. These differences may be responsible for the sensitivity of this subtype to therapy, thus justifying an evaluation of the activity of T in a relatively homogeneous series of U-LM patients. Methods: From April 2000, 56 patients (pts) with advanced disease, previously exposed to a median of 3 chemotherapy lines (range 1–5), received T within an expanded access programme at two European referral institutions for sarcoma. The clinical records were reviewed focusing on response and treatment outcome. Two pts were excluded from the analysis having received only 1 course of T. Median age was 56 yrs (range 29–73), median number of metastatic sites was 2 (range 1–4), the most frequent metastatic site was lung (88%), 24 patients had a local relapse. Results: A total of 252 courses were delivered (median 3, IQR2–6) and 36% of patients received more than 5 courses of T. Fifty-two patients were evaluable for response. A partial response was observed in 11 patients and stable disease in 15, for a PR rate of 21% and a tumor control rate of 50%. The median progression-free survival was 3.6 months (CI95% 2.6–6.7), with 41% of patients free from progression at 6 months. Conclusions: These results compare favourably with other systemic treatments in advanced U-LMS and support their sensitivity to T. This should prompt further studies to prospectively evaluate the efficacy of T in U-LMS and elucidate possible biological predictive factors (e.g. DNA repair protein expression). [Table: see text]
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Jeffers, M., S. Rong, and G. F. Vande Woude. "Enhanced tumorigenicity and invasion-metastasis by hepatocyte growth factor/scatter factor-met signalling in human cells concomitant with induction of the urokinase proteolysis network." Molecular and Cellular Biology 16, no. 3 (March 1996): 1115–25. http://dx.doi.org/10.1128/mcb.16.3.1115.
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Hepatocyte growth factor/scatter factor (HGF/SF) is a pleiotropic effector of cells expressing the Met tyrosine kinase receptor. Although HGF/SF is synthesized by mesenchymal cells and acts predominantly on epithelial cells, we have recently demonstrated that human sarcoma cell lines often inappropriately express high levels of Met and respond mitogenically to HGF/SF. In the present report we show that HGF/SF-Met signalling in the human leiomyosarcoma cell line SK-LMS-1 enhances its in vivo tumorigenicity, an effect for which the mitogenicity of this signalling pathway is likely to play a role. In addition, we found that HGF/SF-Met signalling dramatically induces the in vitro invasiveness and in vivo metastatic potential of these cells. We have studied the molecular basis by which HGFSF-Met signalling mediates the invasive phenotype. A strong correlation has previously been demonstrated between the activation of the urokinase plasminogen activator (uPA) proteolysis network and the acquisition of the invasive-metastatic phenotype, and we show here that HGF/SF-Met signalling significantly increases the protein levels of both uPA and its cellular receptor in SK-LMS-1 cells. This results in elevated levels of cell-associated uPA and enhanced plasmin-generating ability by these cells. These studies couple HGF/SF-Met signalling to the activation of proteases that mediate dissolution of the extracellular matrix-basement membrane, and important property for cellular invasion-metastasis.
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Chong, Tsz Yan, Horace HY Lee, Judy WP Yam, Kin Long Chow, Ho Shing Wong, Ka Yu Tse, Mark R. Bray, Tak Wah Mak, and Philip PC Ip. "Abstract 5394: Uterine leiomyosarcoma with homologous recombination deficiency is highly sensitive to polo-like kinase 4 inhibitor." Cancer Research 82, no. 12_Supplement (June 15, 2022): 5394. http://dx.doi.org/10.1158/1538-7445.am2022-5394.
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Abstract INTRODUCTION: Uterine leiomyosarcoma (LMS) is rare and aggressive and without a well-established cancer-driver. Treatment is mainly surgical because most other therapies have limited benefits. Polo-like kinase 4 (PLK4) is a modulator of centriole duplication that is essential for proper cell division. Its inhibition by the PLK4 inhibitor, CFI-400945 resulted in centriole dysregulation, increased in double-strand DNA damage, and ultimately, mitotic catastrophe and cell death. Recent genomic studies have identified a subset of LMS with homologous recombination (HR) repair deficiency. We hypothesize that HR-deficient LMS is highly sensitive to CFI-400945. METHODS: Gene knockouts (KO) were done by CRISPR-Cas9 system. Plasmids encoding the BRCA1- and BRCA2-targeting guide RNA, Cas9 protein, and selection markers were generated and were transfected into SK-UT-1 and SKN followed by selection with antibiotics. The KO and wild-type cell lines were treated with CFI-400945 in vitro. Percentage apoptosis was evaluated by Annexin V-PI staining and cell proliferation was examined by sulforhodamine B assay. To determine whether BRCA1- and BRCA2-KO had any effects on double-strand DNA damage, the γH2AX foci were studied by immunofluorescence staining. RESULTS: CFI-400945 caused a greater degree of apoptosis and less cell viability in the KO than in the wild-type cell lines. In SK-UT-1, quantity of apoptotic cells for BRCA2-KO and BRCA1-KO were 77% and 34%, respectively, versus 27% in the wild-type cell line (p<0.05). Less dramatic findings were observed in SKN with respective percentages of 20% and 25% versus 15% (p<0.05). In SK-UT-1, the mean IC50 for BRCA2-KO and BRCA1-KO were 3.4 nM and 5.5 nM, respectively, versus 7.8 nM in the wild-type cell line (p<0.05). Similar observations were noted in SKN, with respective mean IC50 of 6.5 nM and 7.1 nM versus 10.0 nM (p<0.05). After CFI-400945 treatment, the γH2AX/Hoechst 33342 staining intensity ratio was 0.40 in SK-UT-1 BRAC2-KO and 0.44 in BRCA1-KO, where both were higher than in the corresponding wild-type cell line with 0.29 (p<0.05). Similarly, the intensities in the two SKN KO cell lines were 0.29 and 0.31, respectively, and higher than in the wild-type cell line with 0.19 (p <0.05). CONCLUSIONS: PLK4 inhibitor induced DNA double-strand breaks in LMS and the repair of these breakages was dependent on HR repair. In vitro, LMS with BRCA1 or BRCA2 deficiencies were more sensitive to the effect of PLK4 inhibitor. Citation Format: Tsz Yan Chong, Horace HY Lee, Judy WP Yam, Kin Long Chow, Ho Shing Wong, Ka Yu Tse, Mark R. Bray, Tak Wah Mak, Philip PC Ip. Uterine leiomyosarcoma with homologous recombination deficiency is highly sensitive to polo-like kinase 4 inhibitor [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5394.
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Young, Aneurin, Edward T. Andrews, James John Ashton, Freya Pearson, R. Mark Beattie, and Mark John Johnson. "Generating longitudinal growth charts from preterm infants fed to current recommendations." Archives of Disease in Childhood - Fetal and Neonatal Edition 105, no. 6 (May 25, 2020): 646–51. http://dx.doi.org/10.1136/archdischild-2019-318404.
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ObjectiveTo use repeated measurements of weight, length and head circumference to generate growth centile charts reflecting real-world growth of a population of very preterm infants with a well-described nutritional intake close to current recommendations.DesignInfants born before 30 weeks gestational age (GA) were recruited. Infants received nutrition according to an integrated care pathway, with nutrient intake recorded daily, weight recorded twice-weekly and length and head circumference weekly. The LMS method was used to construct growth centile charts between 24 and 36 weeks corrected GA for each parameter.SettingA single tertiary neonatal unit in England.Patients212 infants (124 male) (median GA at birth: 27.3 weeks, median birth weight: 900 g).ResultsMedian daily energy, protein, carbohydrate and fat intake were within 3% of published recommendations. The total number of measurements recorded was 5944 (3431 for weight, 1227 for length and 1286 for head circumference). Centile charts were formed for each parameter. Data for male and female infants demonstrated similar patterns of growth and were pooled for LMS analysis. A web application was created and published (bit.ly/sotongrowth) to allow infants to be plotted on these charts with changes in SD score of measurements reported and graphically illustrated.ConclusionsThese charts reflect growth in a real-world cohort of preterm infants whose nutrient intakes are close to current recommendations. This work demonstrates the feasibility of forming growth charts from serial measurements of growing preterm infants fed according to current recommendations which will aid clinicians in setting a benchmark for achievable early growth.
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Sanchez, Guillermo E., Glenn H. Sullivan, and Larry R. Miller. "Impact of an extract from the bran layer of rice on the nutrition of lactating mothers and growth of their exclusively breastfed infants: a longitudinal study." Functional Foods in Health and Disease 12, no. 5 (May 31, 2022): 264. http://dx.doi.org/10.31989/ffhd.v12i5.934.
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Background: The nutritional composition of the bran layer of rice (RB), a globally available product of the rice milling process, has attracted attention as a nutraceutical food source. However, to capitalize on RB’s nutritional properties for humans, it is necessary to achieve a level of bioavailability that increases the efficacy needed to impact the nutritional well-being of lactating mothers and their breastfed infants. To accomplish this, a hydrolyzed enzymatic extract (RBEE) with increased protein, vitamin, carbohydrate, and antioxidant bioactivity has been developed.Objective: To determine the impact of RBEE on the nutritional status of lactating mothers (LM) and the growth of their exclusively breastfed (EBF) infants, living in food-insecure environments in rural Guatemala. Methods: A RBEE daily ration was consumed by post-puerperium LM for 4.5 months of the EBF period. The nutritional impact on LMs was determined by body mass index (BMI). Anemia prevalence and packed cell volume (PCV) were obtained via hematocrit (HCT). Infant growth was determined monthly following World Health Organization (WHO) guidelines, including weight-for-length (WHZ), weight-for-age (WAZ), length-for-age (LAZ), BMI-for-age (BAZ) and head circumference-for-age (HCZ). Initial, midterm and final growth indicators were analyzed by Student t-test for independent samples. Results: While no significant differences were found between the initial and final BMI in LMs, anemia prevalence was significantly reduced from 12.1% to 4%. HCT results in LMs showed a significant increase (p<0.05) in PCV by the study’s end, from 40.55% to 41.42%. Following a student t-test analysis, infant growth indicators WHZ, WAZ and BAZ showed a highly significant (p<0.01) increase. A significant (p<0.05) improvement was detected in WAZ, while LAZ scores improved from -1.9 to -1.76.Conclusions: The dietary supplementation of lactating mothers with an RBEE during the EBF period significantly impacted the mother-infant dyad, with improved growth of the exclusively breastfed infants and a reduction in anemia prevalence in mothers. Further research will include the quantity and quality of the putative increased maternal milk synthesis. Keywords: rice bran, bioactive, bioavailability, enzyme-treated extract, functional food, infants, exclusive breastfeeding, lactating mothers, anemia, chronic, acute, malnutrition, Guatemala, growth, nutrition, children.
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Patetsini, E., B. K. Dimitriadis, and M. Kaloyianni. "Exposure of mussels Mytilus galloprovinciallis to environmental pesticides. Study of LMS, ROS, DNA damage, protein carbonylation and antioxidant capacity for their use as biomarkers." Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology 163 (September 2012): S26. http://dx.doi.org/10.1016/j.cbpa.2012.05.082.
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Johnnie, D. Alwin, Reya Issac, and M. Lakshmi Prabha. "Bio Efficacy Assay of Laccase Isolated and Characterized from Trichoderma viride in Biodegradation of Low Density Polyethylene (LDPE) and Textile Industrial Effluent Dyes." Journal of Pure and Applied Microbiology 15, no. 1 (February 27, 2021): 410–20. http://dx.doi.org/10.22207/jpam.15.1.38.
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This study is based on the biosynthesis of laccase enzyme from fungi Trichoderma viride and its exploitation in biodegradation of polyethylene using Laccase mediator system (LMS -Laccase + 1-HBT) in Low Density Polyethylene (LDPE) and in biodegradation of textile industrial effluent dyes. In different parts of our lifespan we have seen the numerous types of plastics are getting degraded by various methods, still the average time to completely degrade a plastic bottle is at least 450 years.Similarly industry produces over 3.6 thousand individual textile dyes today and utilizes more than 8000 toxic chemicals in numerous methods for textile manufacture comprising dyeing and printing. As polyethylene and textile industrial effluent dyes are causing severe hazardous effect on environment and health issues in all kind of living organisms, it is necessary to degrade plastics and textile industrial effluent dyes in rapid way. Recently researchers have come up with an idea of degrading plastic and textile industrial effluent dyes with the help of microorganisms and enzymes much faster than normal rate. The Laccase enzyme extracted was tested for its optimum temperature and pH. Lowry’s method is used for protein estimation. A control and sample LDPE was subjected to LMS. The tensile strength and elongation of the sample was less than that of the control after 5 days of treatment. This study showed that laccase together with 1-HBT helps to biodegrade polyethylene. The purified laccase enzyme was used for the pretreatment assay and post treatment assay. The Laccase degrades certain reactive dyes like Congo red, Acid Red, Methylene Blue, Brilliant Blue, Metallic Blue and Black. Thus recommends the application of laccase in textile dye colour removal (bioremediation).
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Lorite, María J., Jörg Tachil, Juán Sanjuán, Ortwin Meyer, and Eulogio J. Bedmar. "Carbon Monoxide Dehydrogenase Activity inBradyrhizobium japonicum." Applied and Environmental Microbiology 66, no. 5 (May 1, 2000): 1871–76. http://dx.doi.org/10.1128/aem.66.5.1871-1876.2000.
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ABSTRACT Bradyrhizobium japonicum strain 110spc4 was capable of chemolithoautotrophic growth with carbon monoxide (CO) as a sole energy and carbon source under aerobic conditions. The enzyme carbon monoxide dehydrogenase (CODH; EC 1.2.99.2 ) has been purified 21-fold, with a yield of 16% and a specific activity of 58 nmol of CO oxidized/min/mg of protein, by a procedure that involved differential ultracentrifugation, anion-exchange chromatography, hydrophobic interaction chromatography, and gel filtration. The purified enzyme gave a single protein and activity band on nondenaturing polyacrylamide gel electrophoresis and had a molecular mass of 230,000 Da. The 230-kDa enzyme was composed of large (L; 75-kDa), medium (M; 28.4-kDa), and small (S; 17.2-kDa) subunits occurring in heterohexameric (LMS)2 subunit composition. The 75-kDa polypeptide exhibited immunological cross-reactivity with the large subunit of the CODH of Oligotropha carboxidovorans. The B. japonicum enzyme contained, per mole, 2.29 atoms of Mo, 7.96 atoms of Fe, 7.60 atoms of labile S, and 1.99 mol of flavin. Treatment of the enzyme with iodoacetamide yielded di(carboxamidomethyl)molybdopterin cytosine dinucleotide, identifying molybdopterin cytosine dinucleotide as the organic portion of the B. japonicum CODH molybdenum cofactor. The absorption spectrum of the purified enzyme was characteristic of a molybdenum-containing iron-sulfur flavoprotein.
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Refet-Mollof, Elena, Ouafa Najyb, Rodin Chermat, Audrey Glory, Julie Lafontaine, Philip Wong, and Thomas Gervais. "Abstract B005: Hypoxic sarcoma spheroid on a chip: Insights into treatment response." Clinical Cancer Research 28, no. 18_Supplement (September 15, 2022): B005. http://dx.doi.org/10.1158/1557-3265.sarcomas22-b005.
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Abstract Context: Hypoxia is found in at least 47% of soft-tissue sarcoma (STS) patients and is known to be a major contributor to treatment resistance, metastasis progression and associated with poor prognosis. Indeed, expression of hypoxic proteins (HIF1-α, CAIX) in STS is associated with a shorter overall survival, and a shorter progression-free survival in patients. Specifically, Yang et al. demonstrated in 2017 that patients stratified by a 24-gene hypoxia signature had a poorer distant metastasis free survival. Nevertheless, hypoxia is seldom considered during treatment development due to the lack of a user-friendly way to culture naturally hypoxic 3D tumor models. Therefore, we developed a user-friendly in vitro preclinical tool allowing the study of natural chronic hypoxia and its effect on treatment response. Methods: Microfabrication technologies were used to design a microfluidic chip allowing culture, maintenance, treatment, and analysis of 240 STS naturally hypoxic spheroids (750µm of diameter). SK-LMS-1 Human leiomyosarcoma and STS117 (human primary undifferentiated pleomorphic sarcoma cells) spheroids were formed 48h after seeding. Gold-standard hypoxic protein Hypoxia Inducible Factor 1 alpha (HIF1- α) and Carbonic Anhydrase IX (CAIX) expression was assessed by Western Blot (WB). Spatial distribution of CAIX was assessed by immunofluorescence (IF). Based on the IF, an in silico model of the oxygen consumption of the spheroids was built using COMSOL. As a proof of concept, the spheroids were treated using two oxygen-dependent treatment: RT and hypoxia prodrug Tirapazamine (TPZ). DNA-damages were quantified using yH2AX in IF to assess treatment efficacy. Results: These spheroids on-a-chip are the largest to date (>750µm), and express gold-standard hypoxic protein CAIX in their core only, a feature absent in spheroids of 450µm of diameter of the same cell lines. HIF1-α expression patterns were cell line dependent. CAIX expression in SK-LMS-1 and STS117 large spheroids was respectively 7.8 and 24 times higher than in 450µm spheroids. Quantification of DNA damages (count of yH2AX foci/nuclei area in mm²) demonstrated that TPZ preferentially targets the hypoxic core. However, the response (DNA damages) to RT alone showed no evidence of radioresistance of the hypoxic regions, on samples fixed 24h after irradiation. A possible explanation could be that, in 24h, the oxygen-dependent DNA repair and the accumulation of DNA damages in hypoxic regions hide the oxygen-dependent radioresistance. Therefore, quantification of DNA damages with yH2AX 30min after irradiation is currently being investigated, as maximum DNA damage response has been shown to be observed at this time point. Our 750µm spheroids naturally display a hypoxic core expressing gold-standard hypoxic protein CAIX at 120µm of depth, a feature absent in smaller spheroids. Together, these results cement our microfluidic device and our spheroids as a potent fundamental and pre-clinical tool to explore the biology of hypoxia on 3D tumor model, its effects on treatment response. Citation Format: Elena Refet-Mollof, Ouafa Najyb, Rodin Chermat, Audrey Glory, Julie Lafontaine, Philip Wong, Thomas Gervais. Hypoxic sarcoma spheroid on a chip: Insights into treatment response [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr B005.
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Przybyl, Joanna, Angela Tolwani, Sushama Varma, and Matt van de Rijn. "Abstract B015: Targeting hexosamine biosynthesis pathway for the treatment of desmoid tumors." Clinical Cancer Research 28, no. 18_Supplement (September 15, 2022): B015. http://dx.doi.org/10.1158/1557-3265.sarcomas22-b015.
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Abstract Cancer cells rewire metabolic pathways and energy production to support the enhanced proliferation, invasion and resistance to treatment. The three main glucose metabolism pathways that support growth of cancer cells are: a) the glycolysis pathway for energy production; b) the pentose phosphate pathway for biomass production; and c) the hexosamine biosynthesis pathway (HBP) for protein glycosylation. It is known that the activation of HBP leads to altered glycosylation of oncogenes, transcription factors and kinases in many types of cancer. These aberrations may lead to increased proliferation and survival of tumor cells, and may be associated with resistance to therapy. A better understanding of the role of HBP in malignancies has the potential for clinical implications. Several studies demonstrated that pharmacological inhibition of GFPT2 (glutamine-fructose-6-phosphate transaminase 2, the first and rate-limiting enzyme in HBP) and the enzymes that act downstream of HBP may exhibit anti-tumorigenic effect both in vitro and in vivo, and may modulate sensitivity to chemo-, radio- and immunotherapy. Most of these studies focused on carcinomas and the role of HPB in sarcoma has not been studied. We recently reported a remarkable enrichment of genes involved in HBP in a subset of leiomyosarcoma (LMS) and demonstrated that expression of GFPT2 in LMS is associated with poor clinical outcome. We identified the c-Myc oncoprotein as a potential target of HPB that may be stabilized by aberrant glycosylation in LMS. Here we show the results of a large-scale screening of 260 primary specimens of 33 types of soft tissue lesions. In addition to expression in a subset of LMS, we observed near universal expression of GFPT2 in 34 of 35 desmoid type fibromatosis (DTF), independent of the mutation type of the CTNNB1 gene. Gene Set Enrichment Analysis of a previously published 3SEQ transcriptomic dataset composed of DTF and 9 other types of fibrotic lesions identified significant enrichment of other genes implicated in HBP and multiple glycosylation-associated pathways in DTF compared to the other types of fibrotic lesions. Our analysis identified ATF6 (activating transcription factor 6) as a possible target regulated by aberrant glycosylation as a consequence of HBP activation in DTF. ATF6 is a glycoprotein that has been demonstrated to underlie the resistance to chemotherapy in osteosarcoma, to have a pro-oncogenic role in primary liver cancers and has been proposed as a therapeutic target in cystic fibrosis. Others have shown that targeting HBP can provide therapeutic benefit in a number of preclinical models of carcinoma. Our studies offer new insights into the mechanisms of DTF tumorigenesis and, when confirmed by in vitro studies, will provide a rationale to explore the potential of therapeutic targeting of HBP in DTF. Citation Format: Joanna Przybyl, Angela Tolwani, Sushama Varma, Matt van de Rijn. Targeting hexosamine biosynthesis pathway for the treatment of desmoid tumors [abstract]. In: Proceedings of the AACR Special Conference: Sarcomas; 2022 May 9-12; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2022;28(18_Suppl):Abstract nr B015.
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Lee, S. J., Y. L. Choi, E. J. Lee, B. G. Kim, D. S. Bae, G. H. Ahn, and J. H. Lee. "Increased expression of calpain 6 in uterine sarcomas and carcinosarcomas: an immunohistochemical analysis." International Journal of Gynecologic Cancer 17, no. 1 (January 2007): 248–53. http://dx.doi.org/10.1111/j.1525-1438.2006.00756.x.
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Calpain 6 (Capn6) is one of the calcium-dependent intracellular nonlysosomal proteases. Recently, Capn6 was found to be overexpressed in leiomyosarcomas (LMSs) compared with normal myometrium. This investigation was performed to determine the expression of Capn6 in uterine sarcomas and carcinosarcomas and to determine whether there is a relationship between the clinical findings and the expression of Capn6. Seventeen cases, treated from 1994 to 2004, were evaluated. These included five LMS, seven endometrial stromal sarcomas, and five uterine carcinosarcomas (malignant mullerian mixed tumor [MMMT]). Formalin-fixed, paraffin-embedded tissue sections were immunostained with anti-Capn6 domain-II (anti-DII) and anti-Capn6 domain-T (anti-DT) antibodies. A semiquantitative assessment was performed. All 17 tumors expressed the Capn6 protein; this finding was in contrast to the absence of expression of the Capn6 protein in all of the normal control tissues. The distribution of staining was diffuse. The cytoplasm and nucleus were stained evenly. The mean age of the patients whose samples were stained strongly by anti-DII was higher (P= 0.031). There were no significant associations between tumor stage and staining intensity by anti-DII (P= 1.000) or anti-DT (P= 0.576). However, there was a marginally significant association between tumor subtype and staining intensity (P= 0.054 and P= 0.053, respectively). The expression of Capn6 had no association with disease-free survival (P= 0.367 and P= 0.166, respectively). All of the uterine sarcomas and MMMTs expressed Capn6 protein. This study showed that there were marginally significant associations between tumor subtypes and staining intensity, but no association was found with tumor stage and survival.
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Putluri, Srinivasareddy, and Shaik Yasmin Fathima. "Novel Adaptive Exon Predictor for DNA Analysis Using Singular Value Decomposition." International Journal of Measurement Technologies and Instrumentation Engineering 6, no. 1 (January 2017): 22–32. http://dx.doi.org/10.4018/ijmtie.2017010103.
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This article describes how a realistic prediction of the exon regions in deoxyribonucleic acid (DNA) is a key task in the field of genomics. Learning of the protein coding regions is a key aspect of disease identification and designing drugs. These sections of DNA are known as exons, that show three base periodicity (TBP) which serves as a base for all exon locating methods. Many techniques have been applied successfully, but development is still needed in this area. We develop a novel adaptive exon predictor (AEP) using singular value decomposition (SVD) which notably reduces computational complexity and provides better performance in terms of accuracy. Finally, the exon locating capability of proposed SVD based AEP is tested using a real DNA sequence with accession AF099922, obtained from the National Center for Biotechnology Information (NCBI) database and compared with the existing LMS methods. It was shown that proposed AEP is more efficient for locating the exon regions in a DNA sequence.
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Takei, Hidehiro, Suzanne Powell, and Andreana Rivera. "Concurrent occurrence of primary intracranial Epstein-Barr virus–associated leiomyosarcoma and Hodgkin lymphoma in a young adult." Journal of Neurosurgery 119, no. 2 (August 2013): 499–503. http://dx.doi.org/10.3171/2013.3.jns121707.
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Although Epstein-Barr virus (EBV) infection has been known to be associated with a heterogeneous group of malignancies including Hodgkin lymphoma (HL), its association with smooth-muscle tumors (SMTs) has recently been described. Of these SMTs, a primary intracranial EBV-associated leiomyosarcoma (EBV-LMS) is extremely rare, and most of the reported cases were of immunocompromised and/or pediatric patients. A neurologically asymptomatic, previously healthy 27-year-old man was found to have a PET-positive brain lesion during a staging workup for his recently diagnosed HL. Subsequent MRI revealed a 2.6 × 4.0 × 3.3–cm inhomogeneously enhancing tumor with marked surrounding edema in the right anterior frontal lobe. He was serologically HIV negative. He underwent a right frontal lobectomy with gross-total resection of the tumor. Intraoperatively, the tumor had fairly discrete margins and appeared to arise from the anterior falx (that is, it was dural based). Microscopically, the tumor was composed of interlacing fascicles of spindle cells with brisk mitotic activity and multiple foci of necrosis. Immunohistochemically, the tumor cells were positive for caldesmon and smooth-muscle actin and negative for desmin, CD34, CD99, bcl-2, S100 protein, and GFAP. A Ki-67 labeling index was up to 30%. Epstein-Barr virus–encoded RNA in situ hybridization demonstrated strong diffuse positivity with more than 90% of tumor cells staining. Most of the Reed-Sternberg cells in HL were also labeled with Epstein-Barr virus–encoded RNA. This is the first case of a concurrent occurrence of rare intracranial EBV-LMS and HL in a seemingly “immunocompetent” adult patient (immunocompetence determined by routine laboratory data and clinical history). We should be aware of EBV-SMT as a differential diagnosis of dural-based spindle cell neoplasm in this setting given that patients with HL, even at presentation, exhibit a persistent defect in cellular immunity.
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Ingham, Matthew, Jacob B. Allred, Katherine Gano, Suzanne George, Steven Attia, Melissa Amber Burgess, Sosipatros Alexandros Boikos, et al. "NCI protocol 10250: A phase II study of temozolomide and olaparib for the treatment of advanced uterine leiomyosarcoma." Journal of Clinical Oncology 38, no. 15_suppl (May 20, 2020): TPS11570. http://dx.doi.org/10.1200/jco.2020.38.15_suppl.tps11570.
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TPS11570 Background: Soft tissue sarcoma (STS) is a heterogenous malignancy of mesenchymal origin and includes more than 50 biologically distinct subtypes. Leiomyosarcoma (LMS), a neoplasm of smooth muscle origin, represents up to 20% of STS. The uterus is the most common site of origin in women. Advanced uterine LMS (uLMS) is initially treated with gemcitabine + docetaxel or anthracycline-based chemotherapy but overall survival remains < 24 mos. Besides recurrent alterations in RB1, TP53 and ATRX, insight into cancer biology of uLMS remains limited. Recently, whole exome and transcriptomic sequencing studies suggest uLMS harbors characteristic defects in the homologous recombination (HR) DNA repair pathway and thus features of BRCAness. HR-deficient cancers are unable to efficiently repair double-stranded DNA breaks and appear sensitive to treatment with poly ADP-ribose polymerase (PARP) inhibitors. In preclinical studies, the combination of temozolomide (T), an alkylating agent, and olaparib (O), a PARP inhibitor, was synergistic and markedly suppressed proliferation of uLMS models. A recent phase II study in small cell lung cancer defined the RP2D for T + O where the chief toxicity was myelosuppression. Methods: NCI Protocol #10250 is a single-arm, open-label, multi-center phase II clinical trial of T + O in patients with advanced uLMS. Eligible pts have ECOG PS ≤ 2, progression on ≥ 1 prior line of therapy and disease measurable by RECIST v1.1 and amenable to image-guided biopsy. Pts receive T 75 mg/m2 PO daily + O 200 mg PO BID on days 1-7 in 21-day cycles. The 1° endpoint is objective response rate (ORR). A one-stage binomial design is used to evaluate for an ORR ≤ 10% (null hypothesis) versus ≥ 35% (alternative hypothesis). The design calls for 22 patients. If 5/22 respond, the treatment is promising. This design yields 93% power and 1-sided type I error of 6%. 2° endpoints include progression free survival and safety. All pts undergo tumor biopsies pre-treatment and during cycle 2. Tissue is used for correlative analysis interrogating uLMS for features of BRCAness through (a) whole exome sequencing/RNAseq to evaluate for alterations in HR pathway component genes, (b) RAD51 foci formation by immunohistochemistry as a functional marker of HR pathway activity and (c) protein expression of Schlafen family member number 11 (SLFN11), an emerging biomarker for PARPi. Tumors are also evaluated for MGMT protein expression, a known determinant of sensitivity to T. The study opened to accrual 10/2019. Clinical trial information: NCT03880019.
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Hartner, L. P., L. Rosen, M. Hensley, D. Mendelson, A. P. Staddon, W. Chow, O. Kovalyov, et al. "Phase 2 dose multi-center, open-label study of ARQ 501, a checkpoint activator, in adult patients with persistent, recurrent or metastatic leiomyosarcoma (LMS)." Journal of Clinical Oncology 25, no. 18_suppl (June 20, 2007): 20521. http://dx.doi.org/10.1200/jco.2007.25.18_suppl.20521.
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20521 Background: ARQ 501 selectively induces apoptosis in cancer cells by inducing a rapid and sustained increase of the pro- apoptotic protein E2F-1. ARQ 501 has been studied in three phase 1 studies, demonstrating acceptable toxicity and encouraging signs of efficacy. A 54 y/o female with metastatic LMS who failed 7 previous therapies achieved a prolonged PR on ARQ 501 monotherapy. This was consistent with preclinical data, where induction of E2F-1 and corresponding efficacy in human leiomyosarcoma xenografts was observed. Methods: A phase 2 study in adult LMS patients (>3 prior systemic therapies) was initiated to assess ORR, TTP and further characterize the safety of ARQ 501. ORR included CR, PR and SD=4 mo. Four week cycles (ARQ 501 450mg/m2) were repeated until progression, unacceptable toxicity, or another discontinuation criterion. Results: 49 patients were enrolled and 45 received ARQ 501. Data is available for 43 patients (4M/39F, median age, 54). Of the 43, 10 did not reach a protocol defined tumor assessment (4 deaths, 5 PD and 1 lost to follow-up prior to week 8), 19 have been assessed for response per RECIST at eight weeks (7 SD of 8–28+ weeks, 1 PR, 11 PD) and 14 active patients yet to reach first tumor assessment. The most common AEs were: anemia (68%, 21%=G3), hyperbilirubinemia (35%, 6%=G3), fatigue (35%, 0%=G3), nausea (30%, 0%=G3), constipation (24%), hemolysis (21%, 6%=G3), dyspnea (21%), and vomiting (21%). One treatment related death was reported in a 47 y/o Asian male with severe hemolysis following a single infusion of ARQ 501 at 450 mg/m2. The pt was hospitalized, but severe hemolysis led to acute renal failure and the patient expired after 4 days. Conclusions: ARQ 501 was administered to 45 patients with advanced, recurrent or persistent leiomyosarcoma. Several patients have achieved some clinical benefit (1 PR, 3 prolonged SD), further analysis of efficacy data is warranted prior to additional clinical investigation. No significant financial relationships to disclose.
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Wu, Feitong, Karen Wills, Laura L. Laslett, Brian Oldenburg, Graeme Jones, and Tania Winzenberg. "Associations of dietary patterns with bone mass, muscle strength and balance in a cohort of Australian middle-aged women." British Journal of Nutrition 118, no. 8 (October 9, 2017): 598–606. http://dx.doi.org/10.1017/s0007114517002483.
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AbstractInfluences of dietary patterns on musculoskeletal health are poorly understood in middle-aged women. This cross-sectional analysis from a cohort of 347 women (aged 36–57 years) aimed to examine associations between dietary patterns and musculoskeletal health outcomes in middle-aged women. Diet was measured by the Cancer Council of Victoria FFQ. Total body bone mineral content (TB BMC), femoral neck and lumbar spine bone density (dual-energy X-ray absorptiometry), lower limbs muscle strength (LMS) and balance tests (timed up and go test, step test, functional reach test (FRT) and lateral reach test) were also measured. Exploratory factor analysis was used to identify dietary patterns and scores for each pattern generated using factor loadings with absolute values ≥0·20. Associations between food pattern scores and musculoskeletal outcomes were assessed using multivariable linear regression. Three dietary patterns were identified: ‘Healthy’ (high consumption of a plant-based diet – vegetables, legumes, fruit, tomatoes, nuts, snacks, garlic, whole grains and low intake of high-fat dairy products), ‘high protein, high fat’ (red meats, poultry, processed meats, potatoes, cruciferous and dark-yellow vegetables, fish, chips, spirits and high-fat dairy products) and ‘Processed foods’ (high intakes of meat pies, hamburgers, beer, sweets, fruit juice, processed meats, snacks, spirits, pizza and low intake of cruciferous vegetables). After adjustment for confounders, Healthy pattern was positively associated with LMS, whereas Processed foods pattern was inversely associated with TB BMC and FRT. The associations were not significant after accounting for multiple comparisons. There were no associations with any other outcomes. These results suggest that maintaining a healthy diet could contribute to bone acquisition, muscle strength and balance in adult life. However, while they provide some support for further investigating dietary strategies for prevention of age-related loss of muscle and deterioration in balance, the exploratory nature of the analyses means that confirmation in longitudinal studies and/or trials with pre-specified hypotheses is needed.
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Levitan, Daniel, Viktoriya London, Rodney A. McLaren, Justin David Mann, Ke Cheng, Michael Silver, Kimen Singh Balhotra, Sandra McCalla, and Kristina Loukeris. "Histologic and Immunohistochemical Evaluation of 65 Placentas From Women With Polymerase Chain Reaction–Proven Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Infection." Archives of Pathology & Laboratory Medicine 145, no. 6 (February 17, 2021): 648–56. http://dx.doi.org/10.5858/arpa.2020-0793-sa.
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Context.— Coronavirus disease 2019 (COVID-19) has been shown to have effects outside of the respiratory system. Placental pathology in the setting of maternal severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remains a topic of great interest because earlier studies have shown mixed results. Objective.— To ascertain whether maternal SARS-CoV-2 infection is associated with any specific placental histopathology, and to evaluate the virus's propensity for direct placental involvement. Design.— Placentas from 65 women with polymerase chain reaction–proven SARS-CoV-2 infection underwent histologic evaluation using Amsterdam consensus group criteria and terminology. Another 85 placentas from women without SARS-CoV-2 constituted the negative control group. A total of 64 of the placentas from the SARS-CoV-2–positive group underwent immunohistochemical staining for SARS-CoV-2 nucleocapsid protein. Results.— Pathologic findings were divided into maternal vascular malperfusion, fetal vascular malperfusion, chronic inflammatory lesions, amniotic fluid infection sequence, increased perivillous fibrin, intervillous thrombi, increased subchorionic fibrin, meconium-laden macrophages (M-LMs) within fetal membranes, and chorangiosis. There was no statistically significant difference in prevalence of any specific placental histopathology between the SARS-CoV-2–positive and SARS-CoV-2–negative groups. There was no immunohistochemical evidence of SARS-CoV-2 virus in any of the 64 placentas that underwent staining for viral nucleocapsid protein. Conclusions.— Our study results and a literature review suggest that there is no characteristic histopathology in most placentas from women with SARS-CoV-2 infection. Likewise, direct placental involvement by SARS-CoV-2 is a rare event.
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Dovzhenko, Nadezda Vladimirovna, Victor Pavlovich Chelomin, Andrey Alexandrovich Mazur, Sergey Petrovich Kukla, Valentina Vladimirovna Slobodskova, Aleksandra Anatolievna Istomina, and Avianna Fayazovna Zhukovskaya. "Oxidative Stress in Far Eastern Mussel Mytilus trossulus (Gould, 1850) Exposed to Combined Polystyrene Microspheres (µPSs) and CuO-Nanoparticles (CuO-NPs)." Journal of Marine Science and Engineering 10, no. 5 (May 22, 2022): 707. http://dx.doi.org/10.3390/jmse10050707.
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The ingress of nanoparticles of metal oxides and microfragments of synthetic polymers (microplastics) into a marine environment causes unpredictable consequences. The effects of such particles cannot be predicted due to a lack of ecotoxicological information. In this research, a series of laboratory experiments were conducted on the combined effects of CuO-nanoparticles (CuO-NPs) and polystyrene microspheres (µPSs) on the development of oxidative stress processes in the marine filter-feeder mollusk Mytilus trossulus. Biomarkers of oxidative stress, including the lysosome membrane stability of hematocytes (LMS), the index of antioxidant activity (IAA), the levels of malonaldehyde (MDA) and protein carbonyls (PCs), and DNA damage in digestive gland cells, were measured after 5 days of exposure. Based on a battery of biochemical markers, it was shown that oxidative stress was induced at varying degrees in the experimental mollusks when exposed to CuO-NPs and µPSs both separately and in combination. In contrast, the single-treatment effect on the lysosomal membrane was enhanced by the combined CuO-NPs and µPSs (from 77.14 ± 8.56 to 42 ± 4.26 min). In addition, exposure to both the compounds alone and in combination decreased the IAA (from 22.87 ± 1.25, to 19.55 ± 0.21, 10.73 ± 0.53, and 12.06 ± 1.62 nM/mg protein, respectively). The PC level significantly increased only after CuO-NP exposure (from 0.496 ± 0.02 to 0.838 ± 0.03 μM/mg protein). Furthermore, the results showed that the investigated particles, both alone and in combination, promoted DNA damage in digestive gland cells (from 2.02 ± 0.52 to 5.15 ± 0.37, 18.29 ± 2.14, and 10.72 ± 2.53%, respectively), indicating that these compounds are genotoxic. Overall, the results obtained suggest that oxidative stress is the leading factor in the negative effects of CuO-NPs and µPSs. Considering the exceptional role of genome integrity in the functioning of biological systems, the revealed damages in the DNA molecule structure should be attributed to the most important manifestations of the toxicity of these two forms of marine pollution.
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Muire, Preeti Judith, Lauren H. Mangum, Lee C. Mangum, Gerardo R. Garcia, Desiree R. Romano, and Joseph C. Wenke. "Characterization of the local and systemic temporal changes to the lipid mediator and cytokine cascades following osteotomy and polytrauma." Journal of Immunology 204, no. 1_Supplement (May 1, 2020): 144.3. http://dx.doi.org/10.4049/jimmunol.204.supp.144.3.
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Abstract Polytrauma (PT) inducesglobal changes to inflammatory signaling, contributing to delayed fracturehealing. In a rat model of PT encompassing burn, blunt trauma, and osteotomy, we characterized the local and systemic temporal changes to the lipid mediator(LM) cascade, circulating white blood cells (WBCs), and local levels of growthfactors (GF)s and cytokines. Male rats received either a non-critical 3mmfemoral osteotomy (OST) or PT trauma procedures and were survived for 6, 24,48, 72, 96, 168, or 240hrs and blood, plasma, and fractured femurs werecollected. Whole femurs were homogenized to extract LMs, protein, and RNA, while blood and plasma were used for WBC and LM analysis, respectively. At the fracture site, prostaglandin (PG) E2 was significantly higher in OST at 24 and 72hrs, levels of PGF2α were higher in OST at 24hrs, while resolvin (Rv) D1 was higher in PTat 6hrs post injury. There were no differences in the expression ofcyclooxygenase-2 (COX-2), or lipoxygenase-5 (LOX-5) and LOX-15. In plasma, there were no significant differences in PG levels at any time. Plasma RvE1 was elevated in PT immediately following injury while maresin 1 (MaR1) was significantly lower in PT at 168hrs. Total WBCs and lymphocytes (103cells/uL) were lower in PT overall and significantly so at 72 and 96hrs, respectively. Additionally, the percent of neutrophils in PT was significantlyelevated and percent lymphocytes was decreased at 48 and 168hrs compared toOST. Protein analysis of the fracture site found early elevation of cytokines(IL-1β, IL-13, IL-6) and GFs in PT at 48 and 72hrs. These results indicate thatchanges to circulating LM and WBCs may provide predictive measures of non-unionand therapeutic targets to improve fracture healing.
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Joshi, Pallavi, and Beena Mathur. "Development of value added products from the leaf powders of dehydrated less utilized green leafy vegetables." Nutrition & Food Science 45, no. 2 (March 9, 2015): 302–9. http://dx.doi.org/10.1108/nfs-09-2013-0101.
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Purpose – The purpose of this paper is to analyze the nutritional composition and the acceptability of value-added products prepared from the dehydrated leaf mixture of underutilized green leafy vegetables (GLVs). GLVs are dense in micronutrients and are of great importance to the nutrition of population in developing countries. Nutritive value of commonly consumed GLVs has been studied extensively, but there is limited information available on nutritive value and acceptability of unconventional leafy vegetables. Design/methodology/approach – The nutritional potential and acceptability of leaf mixtures (LMs) prepared from the less-utilized leaves of beet root (Beta vulgaris), carrot (Daucus carota), cauliflower (Brassica oleracea) and turnip (Brassica rapa) which are usually discarded or are used as animal fodder were analyzed in the present study. The LM was prepared by mixing the powders of above-mentioned greens in a definite ratio (1:2:1:1). The LM was analyzed for the proximate, mineral composition (Ca, P, Fe, Cu, Zn, Mn and Mg) and antinutritional factors (oxalate and phenols). In total, 20 different recipes with different levels (0, 5, 10, 15 and 20 per cent) of LM incorporation were prepared and were assessed for quality on the basis of sensory attributes. Findings – The LM contains appreciable amount of proteins, fat, fiber, carbohydrate and calorific value, mineral elements and generally low levels of antinutrients. Products were well-accepted to the level of 10 per cent. Protein, iron and calcium content was significantly (p < 0.05) higher in the LM-incorporated recipes, and the increase was directly proportional to the level of LM incorporated. Originality/value – Dehydrated GLVs are concentrate source of micronutrients and can be used in product formulation. Value addition of traditional products with dehydrated GLVs can be advocated as a feasible food-based approach to combat micronutrient deficiencies.
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Ashley-Martin, Jillian, Regina Ensenauer, Bryan Maguire, and Stefan Kuhle. "Predicting cardiometabolic markers in children using tri-ponderal mass index: a cross-sectional study." Archives of Disease in Childhood 104, no. 6 (January 17, 2019): 577–82. http://dx.doi.org/10.1136/archdischild-2018-316028.
Full textAbstract:
ObjectiveTo model the development of the tri-ponderal mass index (TMI, kg/m3) throughout childhood and adolescence and to compare the utility of the TMI with that of the body mass index (BMI, kg/m2) to predict cardiometabolic risk in a population-based sample of Canadian children and youth.MethodsWe used data from the Canadian Health Measures Survey to model TMI from 6 to 19 years of age. Percentile curves were developed using the LMS method. Logistic regression was used to predict abnormal levels of cardiometabolic markers; predictive accuracy was assessed using the area under the ROC curve (AUC).ResultsMean TMI was relatively stable from ages 6 to 19 years for both sexes, but variability increased with age. There was no notable difference in AUC values for prediction models based on BMI z-score compared with TMI for any of the outcomes. For both BMI z-score and TMI, prediction accuracy was good for homeostasis model assessment insulin resistance and having ≥3 abnormal tests (AUC>0.80), fair for C-reactive protein and poor for the remainder of the outcomes.ConclusionsThe use of a single sex-specific TMI cut-off for overweight or obesity is hampered by the increasing variability of the measure with age. Weight-for-height indices likely have only limited ability to predict cardiometabolic marker levels, and changing the scaling power of height is unlikely to improve predictive accuracy.
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Brígido, Clarisse, Marta Robledo, Esther Menéndez, Pedro F. Mateos, and Solange Oliveira. "A ClpB Chaperone Knockout Mutant of Mesorhizobium ciceri Shows a Delay in the Root Nodulation of Chickpea Plants." Molecular Plant-Microbe Interactions® 25, no. 12 (December 2012): 1594–604. http://dx.doi.org/10.1094/mpmi-05-12-0140-r.
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Several molecular chaperones are known to be involved in bacteria stress response. To investigate the role of chaperone ClpB in rhizobia stress tolerance as well as in the rhizobia-plant symbiosis process, the clpB gene from a chickpea microsymbiont, strain Mesorhizobium ciceri LMS-1, was identified and a knockout mutant was obtained. The ClpB knockout mutant was tested to several abiotic stresses, showing that it was unable to grow after a heat shock and it was more sensitive to acid shock than the wild-type strain. A plant-growth assay performed to evaluate the symbiotic performance of the clpB mutant showed a higher proportion of ineffective root nodules obtained with the mutant than with the wild-type strain. Nodulation kinetics analysis showed a 6- to 8-day delay in nodule appearance in plants inoculated with the ΔclpB mutant. Analysis of nodC gene expression showed lower levels of transcript in the ΔclpB mutant strain. Analysis of histological sections of nodules formed by the clpB mutant showed that most of the nodules presented a low number of bacteroids. No differences in the root infection abilities of green fluorescent protein–tagged clpB mutant and wild-type strains were detected. To our knowledge, this is the first study that presents evidence of the involvement of the chaperone ClpB from rhizobia in the symbiotic nodulation process.
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Kuczak, Michał, and Ewa Kurczyńska. "Cell Wall Composition as a Marker of the Reprogramming of the Cell Fate on the Example of a Daucus carota (L.) Hypocotyl in Which Somatic Embryogenesis Was Induced." International Journal of Molecular Sciences 21, no. 21 (October 30, 2020): 8126. http://dx.doi.org/10.3390/ijms21218126.
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Changes in the composition of the cell walls are postulated to accompany changes in the cell’s fate. We check whether there is a relationship between the presence of selected pectic, arabinogalactan proteins (AGPs), and extensins epitopes and changes in cell reprogramming in order to answer the question of whether they can be markers accompanying changes of cell fate. Selected antibodies were used for spatio-temporal immunolocalization of wall components during the induction of somatic embryogenesis. Based on the obtained results, it can be concluded that (1) the LM6 (pectic), LM2 (AGPs) epitopes are positive markers, but the LM5, LM19 (pectic), JIM8, JIM13 (AGPs) epitopes are negative markers of cells reprogramming to the meristematic/pluripotent state; (2) the LM8 (pectic), JIM8, JIM13, LM2 (AGPs) and JIM11 (extensin) epitopes are positive markers, but LM6 (pectic) epitope is negative marker of cells undergoing detachment; (3) JIM4 (AGPs) is a positive marker, but LM5 (pectic), JIM8, JIM13, LM2 (AGPs) are negative markers for pericycle cells on the xylem pole; (4) LM19, LM20 (pectic), JIM13, LM2 (AGPs) are constitutive wall components, but LM6, LM8 (pectic), JIM4, JIM8, JIM16 (AGPs), JIM11, JIM12 and JIM20 (extensins) are not constitutive wall components; (5) the extensins do not contribute to the cell reprogramming.
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Liang, Zhanwei, Tao Peng, Xueshima Jiao, Yang Zhao, Jie Xie, You Jiang, Bo Meng, Xiang Fang, Xiaoping Yu, and Xinhua Dai. "Latex Microsphere-Based Bicolor Immunochromatography for Qualitative Detection of Neutralizing Antibody against SARS-CoV-2." Biosensors 12, no. 2 (February 7, 2022): 103. http://dx.doi.org/10.3390/bios12020103.
Full textAbstract:
Neutralizing antibody (NAb) is a family of antibodies with special functions, which afford a degree of protection against infection and/or reduce the risk of clinically severe infection. Receptor binding domain (RBD) in the spike protein of SARS-CoV-2, a portion of the S1 subunit, can stimulate the immune system to produce NAb after infection and vaccination. The detection of NAb against SARS-CoV-2 is a simple and direct approach for evaluating a vaccine’s effectiveness. In this study, a direct, rapid, and point-of-care bicolor lateral flow immunoassay (LFIA) was developed for NAb against SARS-CoV-2 detection without sample pretreatment, and which was based on the principle of NAb-mediated blockage of the interaction between RBD and angiotensin-converting enzyme 2. In the bicolor LFIA, red and blue latex microspheres (LMs) were used to locate the test and control lines, leading to avoidance of erroneous interpretations of one-colored line results. Under the optimal conditions, NAb against SARS-CoV-2 detection carried out using the bicolor LFIA could be completed within 9 min, and the visible limit of detection was about 48 ng/mL. Thirteen serum samples were analyzed, and the results showed that the NAb levels in three positive serum samples were equal to, or higher than, 736 ng/mL. The LM-based bicolor LFIA allows one-step, rapid, convenient, inexpensive, and user-friendly determination of NAb against SARS-CoV-2 in serum.