Decellularized skin pretreatment by monophosphoryl lipid A and lactobacillus casei supernatant accelerate skin recellularization.

BACKGROUND: Bioscaffolds and cells are two main components in the regeneration of damaged tissues via cell therapy. Umbilical cord stem cells are among the most well-known cell types for this purpose. The main objective of the present study was to evaluate the effect of the pretreatment of the foreskin acellular matrix (FAM) by monophosphoryl lipid A (MPLA) and Lactobacillus casei supernatant (LCS) on the attraction of human umbilical cord mesenchymal stem cells (hucMSC). METHODS AND RESULTS: The expression of certain cell migration genes was studied using qRT-PCR. In addition to cell migration, transdifferentiation of these cells to the epidermal-like cells was evaluated via immunohistochemistry (IHC) and immunocytochemistry (ICC) of cytokeratin 19 (CK19). The hucMSC showed more tissue tropism in the presence of MPLA and LCS pretreated FAM compared to the untreated control group. We confirmed this result by scanning electron microscopy (SEM) analysis, glycosaminoglycan (GAG), collagen, and DNA content. Furthermore, IHC and ICC data demonstrated that both treatments increase the protein expression level of CK19. CONCLUSION: Pretreatment of acellular bioscaffolds by MPLA or LCS can increase the migration rate of cells and also transdifferentiation of hucMSC to epidermal-like cells without growth factors. This strategy suggests a new approach in regenerative medicine.

The Disruption of Mage-11 Gene via CRISPR/Cas9 Method Induced Apoptosis in the in vitro Model of Prostate Cancer.

BACKGROUNDS AND AIMS: Prostate cancer is the most common malignant cancer among men and is the second deadliest cancer in men after lung cancer. Understanding the molecular mechanisms involved in development and progression of prostate cancer is essential to improve both diagnostic and therapeutic strategies in this regard. In addition, using novel gene therapy-based methods for treatment of cancers has gotten increasing attention during the recent years. Accordingly, this study was aimed to evaluate the inhibitory effect of MAGE-A11 gene, as an important oncogene involved in the pathophysiology of prostate cancer invitro model. The study was also aimed to evaluate the downstream genes related to MAGE-A11. MATERIALS AND METHODS: First, MAGE-A11 gene was knocked out in PC-3 cell line using "Clustered regularly interspaced short palindromic repeats" (CRISPR)/ "CRISPR-associated genes 9" (CRISPR/Cas9) method. Next, the expression levels of MAGE-A11, survivin and Ribonucleotide Reductase Small Subunit M2 (RRM2) genes were determined by quantitative polymerase chain reaction (qPCR) technique. The levels of proliferation and apoptosis were also analyzed in PC-3 cells using CCK-8 and Annexin V-PE/7-AAD assays. RESULTS: The results showed that the disruption of MAGE-A11 by CRISPR/Cas9 method significantly decreased proliferation (P< 0.0001) and enhanced apoptosis (P< 0.05) in PC-3 cells compared to control group. Moreover, the disruption of MAGE-A11 significantly down regulated the expression levels of survivin and RRM2 genes (P< 0.05). CONCLUSION: Our results demonstrated that knocking out MAGE-11 gene by CRISPR/CAS9 technique could efficiently inhibit cell proliferation and induce apoptosis in PC3 cells. Survivin and RRM2 genes might also participated in these processes.

An Effective Method for Decellularization of Human Foreskin: Implications for Skin Regeneration in Small Wounds.

OBJECTIVE: Acellular matrices of different allogeneic or xenogeneic origins are widely used as structural scaffolds in regenerative medicine. The main goal of this research was to optimize a method for decellularization of foreskin for skin regeneration in small wounds. MATERIALS AND METHODS: In this experimental study, the dermal layers of foreskin were divided into two sections and subjected to two different decellularization methods: the sodium dodecyl sulfate method (SDS-M), and our optimized foreskin decellularization method (OFD-M). A combination of non-ionic detergents and SDS were used to decellularize the foreskin in OFD-M. The histological, morphological, and biomechanical properties of both methods were compared. In addition, human umbilical cord mesenchymal stem cells (hucMSCs) were isolated, and the biocompatibility and recellularization of both scaffolds by hucMSC were subsequently determined. RESULTS: We observed that OFD-M is an appropriate approach for successful removal of cellular components from the foreskin tissue, without physical disturbance to the acellular matrix. In comparison to SDS-M, this new bioscaffold possesses a fine network containing a high amount of collagen fibers and glycosaminoglycans (GAG) (P≤0.03), is biocompatible and harmless for hucMSC (viability 91.7%), and exhibits a relatively high tensile strength. CONCLUSION: We found that the extracellular matrix (ECM) structural integrity, the main ECM components, and the mechanical properties of the foreskin are well maintained after applying the OFD-M decellularization technique, indicating that the resulting scaffold would be a suitable platform for culturing MSC for skin grafting in small wounds.

Human Bone Marrow Mesenchymal Stromal Cells Attenuate Tissue Injury and Reduce Inflammation in Experimental Acute Pancreatitis.

Purpose: Acute pancreatitis (AP) which is distinguished by local pancreatic necrosis, followingby systemic organ failure is known as an inflammatory disease. Up to now, there are only a fewtreatment options accessible for patients suffering from AP. In this study, we aimed to examinethe anti-inflammatory capacities of human bone marrow-derived mesenchymal stromal cells(hBM-MSCs) in a detailed AP model experiment. Methods: AP was induced in C57BL/6 mice by intraperitoneal administration of cerulein (100µg/kg/h × 7 doses) at intervals of 1 hour. Then, 2×105 MSCs were infused in the AP mice bytail vein 6 hours after the last cerulein injection. Mice were sacrificed 12 hours following theinjection of hBM-MSC, and blood samples and pancreas tissues were obtained. Results: We first determined the presence of transplanted hBM-MSC in the pancreas of micewith AP, but not the control mice. Our data indicate that administration of hBM-MSCs to micewith AP lead to (i) decreased serum levels of amylase, lipase and myeloperoxidase activities, (ii)downregulation of proinflammatory cytokine, macrophage inflammatory protein 2 (MIP-2), and(iii) upregulation of the anti-inflammatory cytokine, interleukin 10 (IL-10). Moreover, hBM-MSCadministration results in notably attenuated cerulein-induced histopathological alternationsand edema. Conclusion: we demonstrate that hBM-MSC attenuates AP signs and indicating that hMB-MSCtherapy could be a suitable approach for the treatment of inflammatory disease such as AP.

MT2-MMP is differentially expressed in multiple myeloma cells and mediates their growth and progression.

OBJECTIVE: Membrane type-matrix metalloproteinases (MT-MMPs) are known as key regulators of cancer progression/metastasis. However, their roles in the growth and progression of multiple myeloma (MM) have not been yet elucidated. METHODS AND MATERIALS: The expression of 6 MT-MMPs in MM, B cell lines, and normal peripheral blood (PB) cells were measured by RT-PCR, qRT-PCR, flow cytometry, western blotting, and immunocytochemistry. B lymphocytes, CD19-/CD138-, and CD19-/CD138+ cells, known as malignant plasma cells (MPC), were sorted from bone marrow (BM) aspirations of 10 MM patients, and MT2-MMP expression was examined in these cells using qRT-PCR, flow cytometry and immunohistochemistry, and western blotting. Moreover, the expression of MT2-MMP in BM biopsies from 13 normal individuals and 14 MM patients was analyzed by immunohistochemistry. MT2-MMP was also knocked down in U266 cells using siRNA technology and the adhesion, invasion, migration abilities, and cell proliferation were determined and compared with scrambled ones in both in vitro and in vivo studies. RESULTS: Our results showed that MT2-MMP expression is significantly higher in MM cell lines and MPC cells than B cell lines and other PB- or BM-derived cells. MT2-MMP is expressed in BM biopsies from all 14 patients with MM, and 67.85% ± 32.38 of BM cells were positive for MT2-MMP. In contrast, only 0.38 ± 0.76 of BM biopsies from normal individuals were positive for MT2-MMP. Importantly, MT2-MMP was expressed in all the patients' BM biopsies at the diagnosis, but not in the remission phase. MT2-MMP siRNA significantly decreased adhesion, invasion, migration, and 3D cell proliferation of U266 cells. Moreover, in the xenographic model, MT2-MMP siRNA prevented the growth and development of plasmacytoma. Taken together, these data demonstrate that MT2-MMP is strongly expressed in MM cells and plays important role in the growth and progression of these cells, suggesting that MT2-MMP is an appropriate biomarker in diagnosis and therapeutic interventions of MM.

Simultaneous blockade of TIGIT and HIF-1α induces synergistic anti-tumor effect and decreases the growth and development of cancer cells.

PURPOSE: T-cell immunoglobulin and ITIM domain (TIGIT) is an immune checkpoint that is overexpressed on both immune cells and some cancer cells. TIGIT can alter the anti-tumor responses inside the tumor microenvironment. Hypoxia-inducible factor 1-alpha (HIF-1α) plays a significant role in the TME and involves suppressing the anti-tumor responses. Under hypoxic conditions, HIF-1α can enhance the expression of different immune checkpoints. Accordingly, hypoxic TME and TIGIT overexpression cause cancer development. Thus, we decided to inhibit tumor cell expansion by inhibiting TIGIT and HIF-1α molecules and discovering the relationship between TIGIT and HIF-1α. METHODS: In this research, we utilized superparamagnetic iron oxide-based NPs (SPIONs) combined with chitosan lactate (CL) and folic acid (FA) nanoparticles (NPs) loaded with TIGIT-siRNA and HIF-1α- siRNA for suppressing TIGIT and HIF-1α in tumor cells and evaluated the consequences of this treatment strategy on tumor growth, apoptosis, and metastasis. RESULTS: The results showed that cancer cells treated with TIGIT and HIF-1α siRNA-loaded SPIONs-CL-FA NPs, strongly suppressed the TIGIT and HIF-1α expression, colony formation ability, angiogenesis, and the growth rate of cancer cells. CONCLUSIONS: Present data suggest the combination treatment of TIGIT and HIF-1α as a novel treatment strategy against colorectal and breast cancer, but more researches are required to realize the complete role of TIGIT and HIF-1α inside the TME.

Liquid Crystal-Mediated 3D Printing Process to Fabricate Nano-Ordered Layered Structures.

The emergence of three-dimensional (3D) printing promises a disruption in the design and on-demand fabrication of smart structures in applications ranging from functional devices to human organs. However, the scale at which 3D printing excels is within macro- and microlevels and principally lacks the spatial ordering of building blocks at nanolevels, which is vital for most multifunctional devices. Herein, we employ liquid crystal (LC) inks to bridge the gap between the nano- and microscales in a single-step 3D printing. The LC ink is prepared from mixtures of LCs of nanocellulose whiskers and large sheets of graphene oxide, which offers a highly ordered laminar organization not inherently present in the source materials. LC-mediated 3D printing imparts the fine-tuning required for the design freedom of architecturally layered systems at the nanoscale with intricate patterns within the 3D-printed constructs. This approach empowered the development of a high-performance humidity sensor composed of self-assembled lamellar organization of NC whiskers. We observed that the NC whiskers that are flat and parallel to each other in the laminar organization allow facile mass transport through the structure, demonstrating a significant improvement in the sensor performance. This work exemplifies how LC ink, implemented in a 3D printing process, can unlock the potential of individual constituents to allow macroscopic printing architectures with nanoscopic arrangements.

The CCL28 levels are elevated in the serum of patients with irritable bowel syndrome and associated with the clinical symptoms.

BACKGROUND: Inflammation and inflammatory mediators have been proposed to be key players in the pathobiology of Irritable bowel syndrome (IBS. The chemokine CCL28 plays a role in the trafficking of inflammatory cells into mucosal tissues. However, its levels in patients with IBS has not been yet elucidated. METHOD: In this study, the levels of CCL28 were measured in the serum of 41 patients with IBS and 41 age- and gender-matched normal individuals using Elisa. Then, the receiver operating characteristic (ROC) curve was conducted to assess the diagnostic value of CCL28. RESULTS: Our data showed that the levels of CCL28 are significantly elevated in patients with IBS compared to the control donors. Moreover, we observed that the level of CCL28 is associated with many clinical symptoms such as constipation, diarrhea, and abdominal pain. The area under the ROC curve was 0.71 (95% confidential interval, 0.598-0.823), the sensitivity and specificity of CCL28 for the diagnosis of IBS patients were 68.3% and 70.7%, respectively with a cut off of 278.9 ng/mL. CONCLUSIONS: We demonstrated that CCL28 is elevated in patients with IBS and correlates with clinical findings, indicating that CCL28 might be an appropriate biomarker for the diagnosis of IBS; however, further studies are necessary.

Frequency of Growth Differentiation Factor 5 rs143383 and asporin D-repeat polymorphisms in patients with hand and knee osteoarthritis in Kurdistan province, Iran.

AIM: Osteoarthritis (OA) is the most common chronic joint disorder, resulting from the breakdown of joint cartilage. It occurs in the knees, hands, and hips, leading to pain, stiffness, inflammation, and swelling. METHODS: In this study, 100 hand and knee OA patients, meeting the American College of Rheumatology criteria were included in the case group, and 100 healthy individuals were allocated to the control group. Blood samples were collected from the participants. After DNA extraction, genotyping was carried out for GDF5 rs143383 C/T polymorphism by allele-specific polymerase chain reaction (ASPCR) and for D-repeat alleles of asporin (ASPN) by conventional PCR assay. RESULTS: The results showed that the frequency of the D14 allele of ASPN was significantly higher than other alleles in the case group (P = .0001). Also, the frequency of the D14 allele among women was significantly higher than in men (P = .004). Moreover, the frequency of the TT allele in GDF5 rs143383 C/T polymorphism was significantly higher than the CC and CT alleles in the case group, compared with the control group (P = .001). A significant difference was found between the TT allele and other alleles in female and male patients compared with the control group (P = .02). CONCLUSIONS: The D14 allele of the ASPN gene and TT allele of the GDF5 gene (rs143383 + 104T/C) are associated with hand and knee OA in the Kurdish population, indicating that these alleles could be risk factors for OA, at least in our populations.

T-cell immunoglobulin and ITIM domain, as a potential immune checkpoint target for immunotherapy of colorectal cancer.

The importance of the tumor microenvironment in cancer progression has been well studied for many years. Immune checkpoint inhibitors (ICIs) are regarded as potential strategies in enhancing the immune responses in patients with cancer, particularly colorectal cancer (CRC). Notably, CRCs are extraordinarily heterogeneous and mostly are microsatellite-stable (MSS) or cold tumors, which means that the immune response is not usually as strong as that of foreign cells. T-cell immunoglobulin and ITIM domain (TIGIT) is a new immune checkpoint receptor overexpressed inside the CRC tumor-immune microenvironments. Moreover, several studies have shown that TIGIT in combination with other ICIs and/or conventional treatments, can lead to a robust anti-tumor response in CRC. This review looks deep inside TIGIT expression patterns, their various functions, and possible immunotherapy strategies to increase survival rates and decrease immune-related adverse events.

Human umbilical cord mesenchymal stem cells improve morphometric and histopathologic changes of cyclophosphamide-injured ovarian follicles in mouse model of premature ovarian failure.

BACKGROUND: Mesenchymal stem cells have restorative effects on premature ovarian failure (POF). The aim of this study was to investigate the effects of human umbilical cord vein MSCs (hUCV-MSCs) on follicular quantitative parameters and histological changes of ovaries in the cyclophosphamide (CTX)-induced POF in mice. MATERIALS AND METHODS: C57BL/6 mice were divided into three groups (10 mice in each group). In the control group, phosphate-buffered saline (PBS) was injected via tail vein following 15 days injection of PBS intraperitoneally (IP). In the CTX group, CTX was administered IP for 15 days and then PBS was injected via tail vein. In the CTX + hUCV-MSCs group, following CTX administration, single dose of the 1 × 106 of hUCV-MSCs were injected into tail vein. H&E, trichrome and PAS staining as well as TUNEL assay were performed on the ovaries tissue sections. The number of follicles, follicular quantitative parameters and apoptotic index were obtained. The serum levels of estradiol and FSH were measured in the mice. RESULTS: In the CTX + hUCV-MSCs group, degenerative changes were decreased and follicular quantitative parameters increased in the ovarian follicles compared to the CTX group. In this group number of follicles was increased, apoptotic index was decreased, estradiol and FSH levels were decreased and increased, respectively, all of them improved compared to the CTX group. The mean percentage areas of collagen fibers content were decreased compared to the CTX group. CONCLUSION: Results showed that, hUCV-MSCs administration increases follicular quantitative parameters and improve degenerative changes in the follicles following CTX injury.

Platelet Indices and CXCL12 Levels in Patients with Intrauterine Growth Restriction.

BACKGROUND: Intrauterine growth restriction (IUGR) is a multifactorial condition, and the precise mechanism is still unknown. In the current study, we aimed to determine the relationship between the platelet (PLT) indices and CXC12 levels in patients with IUGR. PATIENTS AND MATERIALS: In this study, 36 patients with IUGR and 36 healthy pregnant mothers were enrolled as the case and control groups, respectively. Gestational age for both groups was between 24 and 40 years. Blood samples were taken, and platelet indices were examined by a full-diff cell counter. Serum levels of CXCL12 were measured by ELISA, and the data were analyzed using an independent Student's t-test. RESULTS: In this study, we observed that the mean value of PLT count (154.3 ± 50 vs 236 ± 36) and plateletcrit (0.124 ± 0.038 vs 0.178 ± 0.021) were significantly lower in the case than the control group. In contrast, the mean platelet volume (7.94 ± 0.55 vs 7.62 ± 0.53) and platelet distribution width (17.57 ± 0.7 vs 16.96 ± 0.59) were significantly higher in the case than the control group. More importantly, we found that the serum levels of CXCL12 were significantly higher (5.3 ng/mL± 3.1 vs 2.8 ± 1.6) in the patients compared to the pregnancy controls. CONCLUSION: Our data show that platelet indices are changed in IUGR, and the levels of circulating CXCL12 are increased in patients with IUGR. These findings provide a base for further studies to better defining the pathophysiology of IUGR.

The Clinical Effect of Electroconvulsive Therapy and Its Relationship with Serum Levels of MMP-9 and CXCL12 in Patients with Mania.

INTRODUCTION: Electroconvulsive therapy (ECT) is a non-pharmacological method for the treatment of psychiatric disorders. The precise biochemical mechanism of the effects of ECT is not clear, and since the two factors including matrix metalloproteinase-9 (MMP-9) and stromal cell-derived factor-1 alpha (CXCL12) play an important role in improving nerve damage, the effects of ECT and its relation with serum levels of MMP-9 and CXCL12 in patients with mania were investigated in this study. METHODS: In this before and after intervention study, the patients with mania, referring to the Qods Hospital in Sanandaj, were selected by the census method during the years 2015-2018. Young's test was performed 24 hrs before and after the first, third, and sixth sessions of ECT. For biochemical analysis, 3 mL of peripheral blood were taken prior to any anesthesia and 6 hrs after the first, third, and sixth sessions. Data were analyzed by two-way ANOVA and Pearson correlation coefficient by using the SPSS16 software. RESULTS: The results showed a significant decrease in Young's test scores during the first to the sixth session of ECT (P≤0.05). Although the levels of CXCL12 were slightly increased after the sixth course of ECT, they were not significant. Moreover, there were no significant relationship between the Young's test score and the serum levels of both MMP-9 and CXCL12 (P≥0.05). CONCLUSION: ECT improved patients clinically, but this effect was independent of serum levels of MMP-9 and CXCL12, and possibly other biochemical factors are involved in this pathway.

Relationship between Helicobacter pylori cagA Genotypes Infection and IL-10 and TGFß1 Genes' Expression in Gastric Epithelial Cells.

BACKGROUND: The correlation of Helicobacter pylori infection with gastritis, peptic ulcer, and gastric cancer has been proven. The aim of this study was to determine the effects of cagA + and cagA - genotypes of H. pylori on genes expression of interleukin (IL) -10 and tumor growth factor (TGF) ß1 in gastric epithelial cells of patients with gastritis and H. pylori infection. METHODS: In all, 45 gastric biopsy samples were collected from patients with gastritis and H. pylori infection admitted to Tohid Hospital in Sanandaj city. Status of urease and cagA genes of H. pylori were directly determined from the biopsy samples using polymerase chain reaction (PCR) method. Expression of IL-10 and TGF-ß1 genes in gastric epithelial cells of patients with gastritis and cagA + and cagA- genotypes of H. pylori infection was serveyed using real-time PCR method. RESULTS: Overall, 25 samples had infection with H. pylori cagA + and 20 with cagA - genotypes. This study showed that there is a positive correlation between cagA - genotypes of H. pylori and increasing of IL-10 gene expression in gastric epithelial cells of patients with gastritis (P = 0.001). CONCLUSIONS: Level of gene expression of IL-10 as an anti-inflammatory cytokine in gastric epithelial cells of patients with H. pylori infection is connected to cagA- genotypes.

Hydrogen Peroxide Preconditioning Promotes Protective Effects of Umbilical Cord Vein Mesenchymal Stem Cells in Experimental Pulmonary Fibrosis.

Purpose: Idiopathic pulmonary fibrosis (IPF) is a progressive lung disorder with few available treatments. Mesenchymal stem cell therapy (MSCT), an innovative approach, has high therapeutic potential when used to treat IPF. According to recent data, preconditioning of MSCs can improve their therapeutic effects. Our research focuses on investigating the anti-inflammatory and antifibrotic effects of H2 O2 -preconditioned MSCs (p-MSCs) on mice with bleomycin-induced pulmonary fibrosis (PF). Methods: Eight-week-old male C57BL/6 mice were induced with PF by intratracheal (IT) instillation of bleomycin (4 U/kg). Human umbilical cord vein-derived MSCs (hUCV-MSCs) were isolated and exposed to a sub-lethal concentration (15 µM for 24 h) of H2 O2 in vitro. One week following the injection of bleomycin, 2×105 MSCs or p-MSCs were injected (IT) into the experimental PF. The survival rate and weight of mice were recorded, and 14 days after MSCs injection, all mice were sacrificed. Lung tissue was removed from these mice to examine the myeloperoxidase (MPO) activity, histopathological changes (hematoxylin-eosin and Masson's trichrome) and expression of transforming growth factor beta 1 (TGF-ß1) and alpha-smooth muscle actin (α-SMA) through immunohistochemistry (IHC) staining. Results: Compared to the PF+MSC group, p-MSCs transplantation results in significantly decreased connective tissue (P<0.05) and collagen deposition. Additionally, it is determined that lung tissue in the PF+pMSC group has increased alveolar space (P<0.05) and diminished expression of TGF-ß1 and α-SMA. Conclusion: The results demonstrate that MSCT using p-MSCs decreases inflammatory and fibrotic factors in bleomycin-induced PF, while also able to increase the therapeutic potency of MSCT in IPF.

Construction of A Synthetic Gene Encoding the Multi-Epitope of Toxoplasma gondii and Demonstration of the Relevant Recombinant Protein Production: A Vaccine Candidate.

BACKGROUND: Toxoplasma gondii is a widely-distributed parasite all over the world whose attributed severe afflicting complications in human necessitate the development of serodiagnostic tests and vaccines for it. Immunological responses to monovalent vaccines and the application of diagnostic reagents including single antigens are not optimally effective. Bioinformatics approaches were used to introduce these epitopes, predict their immunogenicity and preliminarily evaluate their potential as an effective DNA vaccine and for serodiagnostic goals. MATERIALS AND METHODS: A 3D structure of proteins was predicted by I-TASSER server, and linear and conformational B cell and T cell epitopes were predicted using the online servers. Then, the predicted epitopes were constructed and called Toxoeb, and their expression in the prokaryotic and eukaryotic cells was demonstrated using SDS-PAGE. In the next step, Western blotting with pooled sera of mice infected with T. gondii was done. RESULTS: The current in silico analysis revealed that the B cell epitopes with high immunogenicity for GRA4 protein were located in the residues 34-71, and 230-266, for GRA14 in 308-387, for SAG1 in 182-195, 261-278, and for GRA7 in residues 101-120, 160-176. The T cell epitopes were selected in overlapping regions with the B cell epitopes. The immunogenic region for GRA4 are in the residues 245-253, 50-58, and 40-54, for GRA14 in 307-315, 351-359, and 308- 322, for SAG1 261-269, and 259-267, and for GRA7 in the residues 103-112, and 167-175. The results of the western blotting showed that the expressed protein had immunogenicity. CONCLUSION: Our constructed multi-epitope of T. gondii could be considered as a candidate for diagnostic and vaccination purposes.

Designing new generation of potent inhibitors against membrane-type matrix metalloproteinase-2: a computational effort against multiple myeloma.

Matrix metalloproteinases (MMPs) play important roles in cancer progression and, despite their inhibitors have failed in the clinical trials, they have always been considered as suitable targets for the treatment of tumor. We have recently shown that membrane type (MT) 2-MMPs, is selectively expressed in multiple myeloma (MM) cells and mediates the metastatic characteristics of these cells. In this study, we designed efficient inhibitors against MT2-MMP using state-of-art molecular modeling methods. First, the 3D structure of MT2-MMP was predicted. Then, the proposed potent inhibitors against two regions of the catalytic domain of MT2-MMP (active site and MT-LOOP) were identified through molecular docking, QM-MM and molecular dynamics simulations from a set of compounds in Analyticon library, IBS library, Maybridge screening fragment library and drugbank library. Moreover, ADME estimation showed that pharmacokinetic properties of inhibitors are in the acceptable range for humans. Finally, our data suggested that compounds 'structures.722' (dobutamine) and 'M2' are suitable candidates to inhibit MT2-MMP for further examination in the laboratory.Communicated by Ramaswamy H. Sarma.

Association of CYP2C19 and HSP70 Genes Polymorphism with Aspirin- Exacerbated Respiratory Disease in a Kurd Population.

BACKGROUND: CYP2C19 a metabolizing enzyme and Heat Shock Proteins (HSP) are induced in stress conditions, such as hypoxia and ischemia. Recently, polymorphism in the CYP2C19 and HSP genes has been established in Aspirin-Exacerbated Respiratory Disease (AERD). OBJECTIVE: We investigated the polymorphism of these two genes in Kurdish patients with AERD. METHODS: This study involved 306 subjects, referred to the Be'sat hospital in Kurdistan Province, which were divided into three groups, (i) Aspirin Induced Asthma (AIA), (ii) Aspirin Tolerant Asthma (ATA), and (iii) healthy subjects as control. The subjects as control and ATA\AIA groups were verified by the physician. The demographic data of each subject with respect to age, sex, parental education, and residence was collected. Spirometry was performed on subjects and blood samples were collected for serum Immunoglobulin E (IgE) estimation and molecular tests. Genotyping was done for CYP2C19 681G>A، CYP2C19 636G>A, and HSPA1B1267A>G by using PCR- Restriction Fragment Length Polymorphism (RFLP) and for HSPA1B-179C>T by High Resolution Melting (HRM). RESULTS: Demographic statistics were not significantly different between the three groups (p>0.05). Further, genotypes were also not observed to be significantly different in the genes of CYP2C19 681G>A, CYP2C19 636G>A and HSPA1B1267A>G (p>0.05). However, the heterozygote genotype in HSPA1B-179 C>T in AIA group was higher than the control group (p<0.05). Notably, 92.8 % of the subjects showed heterozygote genotype in HSPA1B1267 A>G. In clinical tests, FEV-1, FVC, and asthma severity in the AIA group were higher than control and additionally IgE levels were lower in this group (p<0.05). CONCLUSION: The results confirm the association of polymorphism in the HSPA1B-179C>T and HSPA1B1267A>G with AERD in the Kurdish population.

Prevalence of Osteosarcopenia and Its Association with Cardiovascular Risk Factors in Iranian Older People: Bushehr Elderly Health (BEH) Program.

Osteosarcopenia is an increasingly recognized geriatric syndrome with a considerable prevalence which increases morbidity and mortality. Although osteosarcopenia is a result of age-related deterioration in muscle and bone, there are many risk factors that provoking osteosarcopenia. These risk factors should be considered by the clinicians to treat osteosarcopenia. We assessed the link between osteosarcopenia and conventional risk factors of cardiovascular diseases. This study was a cross-sectional study that has been conducted within the framework of Bushehr Elderly Health (BEH) program stage II in which participants aged ≥ 60 years were included. Osteopenia/osteoporosis was defined as a t-score ≤ - 1.0 standard deviation below the mean values of a young healthy adult. We defined sarcopenia as reduced skeletal muscle mass plus low muscle strength and/or low physical performance. Osteosarcopenia was considered as the presence of both osteopenia/osteoporosis and sarcopenia. We estimated the age-standardized prevalence of osteosarcopenia for men and women, separately. Using modified Poisson regression analysis, adjusted prevalence ratio (PR) with 95% CI was used to show the measure of associations in the final model. Among 2353 participants, 1205 (51.2%) were women. Age-standardized prevalence of osteosarcopenia was 33.8 (95% CI 31.0-36.5) in men and 33.9 (30.9-36.8) in women. In both sexes, the inverse association was detected with body mass index and having osteosarcopenia (PR 0.84, 95% CI 0.81-0.88 in men and 0.77, 95% CI 0.74-0.80 in women). In both sexes, high-fat mass was positively associated with osteosarcopenia [PR 1.46 (95% CI 1.11-1.92) in men, and 2.25 (95% CI 1.71-2.95) in women]. Physical activity had a significant inverse association in men (PR = 0.64, 95% CI 0.46, 0.88), but not in women. Diabetes was also showed a direct association with osteosarcopenia in men (PR 1.33, 95% CI 1.04-1.69). No associations were detected between the lipid profiles and osteosarcopenia. Results demonstrated a high prevalence of osteosarcopenia in both sexes suggesting a high disease burden in a rapidly aging country. Lifestyle and socioeconomic factors, as well as chronic diseases, were significantly associated with osteosarcopenia.