Structure, function and regulation of versican:the most abundant type of proteoglycan in the extracellular matrix

One of the main members of the large aggregating proteoglycans (PGs) family is versican which is able to bind to hyaluronate. Versican is a chondroitin sulfate proteoglycan and is a key ingredient of the extracellular matrix.  Due to its widespread expression in the body, versican is involved in cell adhesion, proliferation and migration. Induced expression of versican is often observed in tissues such as breast, brain, ovary, gastrointestinal tract, prostate, and melanoma. In addition, versican has important role in development. For example, versican conducts the embryonic cell migration which is essential in the formation of the heart and outlining the path for neural crest cell migration. Several studies in the past decade up to now have shown that versican produced by mononuclear cells has an important role in wound healing and blood vessel formation and suggested that it promotes tumorigenesis and angiogenesis. In this mini-review, we summarise and discuss the role of versican in healthy and pathological tissues and suggest the possible function of transcription factors and signalling pathway in regulation of versican

Evaluation of the Enzymatic Activity of Soluble CD13/APN and CD26/DPP4 in Serum and Urine Samples of Mice with Experimental Autoimmune Encephalomyelitis

Background: Dipeptidyl Peptidase IV (DPP4/CD26) and Amino Peptidase N (APN/CD13) have essential roles in inflammatory diseases. The current study aimed to determine changes in APN and DPP4 enzyme activity in the serum and urine of mice with experimental autoimmune encephalomyelitis (EAE). Materials and Methods: In the present study, female C57BL/6 mice were studied in two groups (control and EAE). Twenty-sevendays after induction, the enzymatic activity of APN and DPP4 in urine and serum samples was measured using a spectrophotometric assay. Results: The enzyme activity of DPP4 was higher in serum and urine of EAE mice than in the control group (mean in serum: 1.04 ± 0.13 pmol/mL and 0.80 ± 0.12 pmol/mL, respectively, P=0.015; mean in urine: 0.26 ± 0.04 pmol/mL and 0.19 ± 0.04 pmol/mL, respectively, P=0.015). However, the enzymatic activity of APN in serum and urine of mice with EAE when compared to the control group had no significant difference (mean in serum: 9.20 ± 1.15 unit/mL and 10.25 ± 1.21 unit/mL, respectively, P=0.132, mean in urine: 0.23 ± 0.27 unit/mL and 0.15 ± 0.05 unit/mL, respectively, P=0.310). Conclusion: The increased DPP4 activity along with normal APN activity in urine and serum samples can be used as an indicator to detect or follow up on the course of MS disease. Confirmation of this finding needs further investigation

Mechanisms of hypoxic up-regulation of versican gene expression in macrophages

Hypoxia is a hallmark of many pathological tissues. Macrophages accumulate in hypoxic sites and up-regulate a range of hypoxia-inducible genes. The matrix proteoglycan versican has been identified as one such gene, but the mechanisms responsible for hypoxic induction are not fully characterised. Here we investigate the up-regulation of versican by hypoxia in primary human monocyte-derived macrophages (HMDM), and, intriguingly, show that versican mRNA is up-regulated much more highly (>600 fold) by long term hypoxia (5 days) than by 1 day of hypoxia (48 fold). We report that versican mRNA decay rates are not affected by hypoxia, demonstrating that hypoxic induction of versican mRNA is mediated by increased transcription. Deletion analysis of the promoter identified two regions required for high level promoter activity of luciferase reporter constructs in human macrophages. The hypoxia-inducible transcription factor HIF-1 has previously been implicated as a key potential regulator of versican expression in hypoxia, however our data suggest that HIF-1 up-regulation is unlikely to be principally responsible for the high levels of induction observed in HMDM. Treatment of HMDM with two distinct specific inhibitors of Phosphoinositide 3-kinase (PI3K), LY290042 and wortmannin, significantly reduced induction of versican mRNA by hypoxia and provides evidence of a role for PI3K in hypoxic up-regulation of versican expression

Studies on regulation of versican gene expression by hypoxia in primary human macrophages

Hypoxia is a hallmark of many pathological tissues. Macrophages accumulate in hypoxic sites and up-regulate a number of hypoxia-inducible genes. The extracellular matrix glycoprotein versican has recently been identified as one such gene, but the mechanisms responsible for hypoxic induction are not well characterised. Here, hypoxic up-regulation of versican was investigated in primary human monocyte-derived macrophages. Flow cytometry of isolated peripheral blood mononuclear cells demonstrated a three-fold increase in versican protein in macrophages after 5 days incubation in hypoxia. Subset analysis showed that macrophages, and not lymphocytes, are the main peripheral blood mononuclear cells which express, and show hypoxic up-regulation of, versican protein and mRNA. This study showed that versican mRNA is up-regulated 34-fold after exposure of primary human macrophages to hypoxia for 18hrs. Further investigation showed that versican mRNA decay rates are not affected by hypoxia, indicating that hypoxic induction of versican mRNA is mediated by increased promoter activity rather than increased mRNA stability. Extensive deletion and transfection analysis of proximal versican promoter luciferase reporter constructs identified two regions which are required for high level activity of the promoter in hypoxic primary human macrophages. A recent publication suggested that hypoxic induction of versican mRNA in macrophages is mediated by the hypoxia inducible transcription factor HIF-1α. Here, bacterial lipopolysaccharide and the hypoxia mimetic agents desferrioxamine and cobalt chloride, three stimuli which are known to induce HIF-1α, were used to investigate the role of HIF-1 in the up-regulation of versican mRNA. Neither LPS nor cobalt chloride caused up-regulation of versican mRNA, although control HIF-1 regulated genes were up-regulated, suggesting that high-level transcription of the versican promoter in hypoxia occurs via a HIF-1 independent mechanism. Lastly, two specific inhibitors of PI3-kinase, LY294002 and Wortmannin, were shown to down-regulate hypoxic induction of versican mRNA, suggesting a possible role for PI3-kinase

Studies on regulation of versican gene expression by hypoxia in primary human macrophages

Hypoxia is a hallmark of many pathological tissues. Macrophages accumulate in hypoxic sites and up-regulate a number of hypoxia-inducible genes. The extracellular matrix glycoprotein versican has recently been identified as one such gene, but the mechanisms responsible for hypoxic induction are not well characterised. Here, hypoxic up-regulation of versican was investigated in primary human monocyte-derived macrophages. Flow cytometry of isolated peripheral blood mononuclear cells demonstrated a three-fold increase in versican protein in macrophages after 5 days incubation in hypoxia. Subset analysis showed that macrophages, and not lymphocytes, are the main peripheral blood mononuclear cells which express, and show hypoxic up-regulation of, versican protein and mRNA. This study showed that versican mRNA is up-regulated 34-fold after exposure of primary human macrophages to hypoxia for 18hrs. Further investigation showed that versican mRNA decay rates are not affected by hypoxia, indicating that hypoxic induction of versican mRNA is mediated by increased promoter activity rather than increased mRNA stability. Extensive deletion and transfection analysis of proximal versican promoter luciferase reporter constructs identified two regions which are required for high level activity of the promoter in hypoxic primary human macrophages. A recent publication suggested that hypoxic induction of versican mRNA in macrophages is mediated by the hypoxia inducible transcription factor HIF-1α. Here, bacterial lipopolysaccharide and the hypoxia mimetic agents desferrioxamine and cobalt chloride, three stimuli which are known to induce HIF-1α, were used to investigate the role of HIF-1 in the up-regulation of versican mRNA. Neither LPS nor cobalt chloride caused up-regulation of versican mRNA, although control HIF-1 regulated genes were up-regulated, suggesting that high-level transcription of the versican promoter in hypoxia occurs via a HIF-1 independent mechanism. Lastly, two specific inhibitors of PI3-kinase, LY294002 and Wortmannin, were shown to down-regulate hypoxic induction of versican mRNA, suggesting a possible role for PI3-kinase.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Frequency of Cutaneous Leishmaniasis and Complete Ulcer Healing in Patients Referred to Skin Diseases and Leishmaniasis Research Center, Isfahan, Iran from 2018 to 2019

Introduction: Iran is one of the most important hot spots for cutaneous leishmaniasis (CL) in the world. To date, no studies have been done on both epidemiological aspects along with the length of the treatment course of CL. This study aimed to determine the relative frequency of CL in patients with suspected skin lesions and the duration of healing after treatment with different regimens. Methods: This cross-sectional study was conducted on patients with CL referred to the skin diseases and leishmaniasis research center (SDLRC) in Isfahan during the years 2018 to 2019. Among 389 patients with suspected skin lesions, 150 cases were included with proven CL. Information such as age, sex, education, location, size of the lesion, duration of treatment, and the rate of recovery were recorded. SPSS software version 20 was used for data analysis, the chi-square, Fisher´s Exact, and one Way ANOVA tests were used with a significant level of p < 0.05. Results: Among 350 admitted cases, 150 cases were CL. positive (42.85%). The rate of complete recovery was higher in cases with an average age of 33.55 ±18.9 years, but these differences were not statistically significant (P =0.077). There was 34 cases more than the other groups in this range of age. ( The rate of complete recovery in patients with a history of migration to endemic areas was higher than in patients without a history of migration (P = 0.81)). The rate of complete recovery in patients whose means treatment duration was 59.03 ± 41.43 days was higher than other recovery periods (P = 0.23). Conclusion: The rate of complete recovery was higher in adult cases than the other groups. In this study, it was proved that the rate of recovery of patients had the significant relationship with the average duration of treatment

Aberrant promoter hypermethylation of miR-335 and miR-145 is involved in breast cancer PD-L1 overexpression

Abstract PD-L1 is one of the most important immune checkpoint molecules in breast cancer that plays an important role in suppressing the immune system when confronted with tumor cells and is regulated by various microRNAs. Among them, microRNA-335-3p and microRNA-145-5p, regulated by DNA methylation, have tumor suppressor activities. We studied the role of miR-335 and -145 on PD-L1 suppression in breast cancer. The expression of miR-355 and miR-145 was significantly downregulated in BC tissues and cell lines compared to their controls, and their downregulation was negatively correlated with PD‐L1 overexpression. In-silico and luciferase reporter systems confirmed that miR-335 and -145 target PD-L1. In BC tissues and cell lines, cancer-specific methylation was found in CpG-rich areas upstream of miR-335 and-145, and up-regulation of PD-L1 expression was connected with hypermethylation (r = 0.4089, P = 0.0147, and r = 0.3373, P = 0.0475, respectively). The higher levels of miR-355 and -145 in BC cells induced apoptosis, arrested the cell cycle, and reduced proliferation significantly. In summary, we found that miR-335 and -145 are novel tumor suppressors inactivated in BC, and these miRs may serve as potential therapeutic targets for breast cancer treatment

In-silico design and evaluation of an epitope-based serotype-independent promising vaccine candidate for highly cross-reactive regions of pneumococcal surface protein A

Abstract Background The pathogenicity of pneumococcus with high morbidity, mortality, and multi-drug resistance patterns has been increasing. The limited coverage of the licensed polysaccharide-based vaccines and the replacement of the non-vaccine serotypes are the main reasons for producing a successful serotype-independent vaccine. Pneumococcal surface protein A (PspA) is an extremely important virulence factor and an interesting candidate for conserved protein-based pneumococcal vaccine classified into two prominent families containing five clades. PspA family-elicited immunity is clade-dependent, and the level of the PspA cross-reactivity is restricted to the same family. Methods To cover and overcome the clade-dependent immunity of the PspAs in this study, we designed and tested a PspA1-5c+p vaccine candidate composed of the highest immunodominant coverage of B- and T-cell epitope truncated domain of each clade focusing on two cross-reactive B and C regions of the PspAs. The antigenicity, toxicity, physicochemical properties, 3D structure prediction, stability and flexibility of the designed protein using molecular dynamic (MD) simulation, molecular docking of the construct withHLADRB1*(01:01) and human lactoferrin N-lop, and immune simulation were assessed using immunoinformatics tools. In the experimental section, after intraperitoneal immunization of the mice with Alum adjuvanted recombinant PspA1-5c+p, we evaluated the immune response, cross-reactivity, and functionality of the Anti-PspA1-5c+p antibody using ELISA, Opsonophagocytic killing activity, and serum bactericidal assay. Results For the first time, this work suggested a novel PspA-based vaccine candidate using immunoinformatics tools. The designed PspA1-5c+p protein is predicted to be highly antigenic, non-toxic, soluble, stable with low flexibility in MD simulation, and able to stimulate both humoral and cellular immune responses. The designed protein also could interact strongly with HLADRB1*(01:01) and human lactoferrin N-lop in the docking study. Our immunoinformatics predictions were validated using experimental data. Results showed that the anti-PspA1-5c+p IgG not only had a high titer with strong and same cross-reactivity coverage against all pneumococcal serotypes used but also had high and effective bioactivity for pneumococcal clearance using complement system and phagocytic cells. Conclusion Our findings elucidated the potential application of the PspA1-5c+p vaccine candidate as a serotype-independent pneumococcal vaccine with a strong cross-reactivity feature. Further in-vitro and in-vivo investigations against other PspA clades should be performed to confirm the full protection of the PspA1-5c+p vaccine candidate

rs12329760 Polymorphism in Transmembrane Serine Protease 2 Gene and Risk of Coronavirus Disease 2019 Mortality

The protease produced by the transmembrane serine protease 2 (TMPRSS2) gene enhances viral infections and has been linked to severe acute respiratory syndrome coronavirus 2 pathogenesis. Therefore, this study evaluated the association between TMPRSS2 and coronavirus disease 2019 (COVID-19) mortality. TMPRSS2 rs12329760 polymorphism was genotyped using the tetraprimer amplification refractory mutation system-polymerase chain reaction method in 592 dead and 693 improved patients. In the current study, the frequency of TMPRSS2 rs12329760 CC than TT genotypes was significantly lower in improved patients than in dead patients. According to the findings of the multivariate logistic regression test, higher levels of mean age, creatinine, erythrocyte sedimentation rate, C-reactive protein, aspartate aminotransferase, lower levels of 25-hydroxyvitamin D, uric acid, and real-time PCR Ct values and TMPRSS2 rs12329760 CC genotype were observed to be associated with increased COVID-19 mortality rates. In conclusion, the TMPRSS2 rs12329760 CC genotype was a polymorphism linked to a significantly higher incidence of severe COVID-19. Further studies are required to corroborate the obtained findings