The PPARγ Agonist Rosiglitazone Is Antifibrotic for Scleroderma Lung Fibroblasts: Mechanisms of Action and Differential Racial Effects

We present novel data demonstrating that the expression of PPARγ is reduced in lung fibroblasts from black SSc-ILD patients as compared to white patients. Activating PPARγ with the agonist rosiglitazone increased the expression of MMP-1 and inhibited collagen type I in lung fibroblasts isolated from white, but not black, SSc-ILD patients. Blocking the c-Met receptor abolishes rosiglitazone's effects on collagen and MMP-1 in lung fibroblasts isolated from white SSc-ILD patients, while augmenting the expression of the c-Met receptor in fibroblasts from black SSc-ILD patients replicates the effects of rosiglitazone seen in whites. We conclude that PPARγ agonists warrant consideration as potential antifibrotic drugs in patients with SSc-ILD. Differential therapeutic effects might be anticipated especially relative to racial differences and the functional expression of the c-Met receptor

Investigate to find common gene and design a PPI network for vector borne diseases (Malaria, Dengue and Chikungunya) – A bioinformatics approach

Malaria, Dengue and Chikungunya are the most common mosquito-borne viral diseases transmitted to humans by day-biting Aedes aegypti and Aedes albopictus mosquitoes. Different types of gene are responsible for these viruses. The principal study of this research is to find the relationship between genetic variant for these three diseases and to create a common pathway regulatory or Protein-Protein Interaction (PPI) network. Our investigation goes through preprocessing, filtering, sorting and gene mining on the gathered gene (Malaria, Dengue and Chikungunya) using R to find the common associated genes by the process of reduction. The investigation shows that about 60% of the collected gene from different standard gene database is responsible for animal virus attack. After preprocessing, filtering and sorting using R toolkit, the number of collected gene for three diseases(A=malaria, B=dengue and C=chikunguniya) is reduced to 35%. Gene mining is done by intersection operation on (A, B), (B, C) and (C, A) that reduces the common associated gene from 35% to 5%. Finally, the reduction is done by intersecting AB, BC and CA that reduces the common gene from 5% to less than 1%. We have discovered five (5) common associated genes for these three virus diseases. However a common pathway with the five (5) common associated genes that has been designed for selective diseases

Potential Therapeutic Effects of a 10 Amino Acid Peptide, M10, in Scleroderma Associated Interstitial Lung Disease (SSc-ILD)

Background: SSc-ILD is a complicated autoimmune disease and a major cause of mortality among scleroderma patients; TGFβ is a crucial profibrotic regulator. We previously demonstrated that a 10 amino acid peptide, M10, reduced collagen production and attenuated fibrosis in a preventive mouse model. In this study, we investigated therapeutic potentials of M10 in a bleomycin-induced fibrosis model in mice and its antifibrotic mechanisms in primary lung fibroblasts. Methods: Efficacy and pharmacokinetics (PK) of M10 were studied in C57BL/6J mice. Severity of fibrosis was evaluated by analyzing the lung mechanics, histology, and soluble collagen content. Concentration of M10 in plasma was estimated by ELISA. Expressions of extracellular matrix (ECM) proteins (e.g. collagen, fibronectin, and tenascin) were measured by immunoblotting and real-time PCR. Cytosolic Ca2+ was measured by the FLIPR Tetra cellular screening system. Expression and activity of matrix metalloproteinases (MMPs) were determined by human MMP antibody array, ELISA and fluorometric assays. Statistical analysis was performed using GraphPad Prism 7 software. Results: M10 significantly reduced lung fibrosis as measured by the Ashcroft score (p \u3c 0.003) and soluble collagen content (p \u3c 0.02) in the therapeutic mouse model. The maximum concentration of M10, 4.8 μg/ml, was achieved within 6 hours of M10-administration, and declined to a minimal level of 0.2 μg/ml by 48 hours. The experimentally obtained half-life of M10 (6.8 ± 0.7 hour) was comparable to the theoretically predicted half-life of M10 (7.2 hour) by ProtLifePred software. In lung fibroblasts, M10 slightly lowered ECM proteins on the mRNA level, significantly reduced collagen on the protein level, and regulated expressions of MMPs in a cell type-dependent manner. In normal lung fibroblasts, in the presence of TGFβ, M10 significantly induced expression of MMP-10 in a Ca2+-independent way. In SSc-ILD lung fibroblasts, M10 significantly reduced expression of MMP-3 in Ca2+-dependent way. In both, normal and SSc-ILD lung fibroblasts, in the presence of M10, TGFβ-mediated Ca2+ induction was significantly (p \u3c 0.001) reduced. Conclusion: M10 exerts convincing stability and antifibrotic effects in in vivo models. M10 reduces ECM proteins, inhibits TGFβ-mediated Ca2+ efflux, and regulates MMP expression in lung fibroblasts. These data suggest that M10 possesses promising potential to reduce TGFβ-mediated outcomes in SSc-ILD

3D Printing and Product Design For Plastic Snap-Fits and Hinges

The aim of this thesis project is to design and 3D print plastic snap-fit and hinges boxes which are used in daily life. All the design was done by using SolidWorks 2021 and 3D printing was successfully done by using the Creality Ender-3 printer and PLA plastics. SolidWorks FEA simulation was used to analyze the stress of the snap-fits and hinges. The stress results found from the simulation were quite accurate compared to theoretical calculations of the cantilever snap-fit box with hinges as the error was only 11%. Whereas cantilever snap-fit and annular snap-fit have errors of 56% and 83% respectively. The maximum stress of the cantilever snap-fit box and cantilever snap-fit box with hinges in simulation results are 127.964 MPa and 90.91 MPa. And the theoretical result is 81.75 MPa. Moreover, the results of the annular snap-fit in simulation and theoretical stress analysis are 0.775 MPa, and 0.422 MPa respectively. Therefore, in the printing parts, there was some uneven area in the cantilever snap-fit box hinges as support used while printing. Otherwise, the printing process of the cantilever snap-fit box and the annular snap-fit box was successful. There are a few types of snap-fit which are cantilever, annular and torsional. In this thesis project, cantilever and annular snap-fit were presented. On the other hand, hinges analysis has also been presented. Though there have quite a lot of errors in theoretical calculations one can easily acknowledge from this research about different types of snap-fits and hinges, 3D printing, and analytical ways of calculating the stress and mass of any material

Analyses Implementation Realities of Legal Frameworks for Sustainable Management of Tanguar Haor Fisheries Resources in Bangladesh

Tanguar Haor (TH) is considered one of the Ecologically Critical Areas (ECAs) of Bangladesh and is internationally recognized as RAMSAR wetland (2nd Ramsar site) known to provide multiple ecosystem services to the society. Nevertheless, multidimensional threats and stressors, the capacity to supply ESs, and the biodiversity of the TH significantly degrades and threatens this wetland's conservation and sustainability. Although the legal framework promises the sustainable conservation of fisheries resources, information on the implementation scenarios of fisheries laws, regulations, and policies in the TH Ramsar are scant. By merging qualitative and quantitative data of primary and secondary sources, this research aimed to analyze the legal framework to check the effectiveness of regulations for non-conflicting fisheries resources and the sustainable conservation of the TH Ramsar. Primary empirical data were collected by employing Participatory Rural Appraisal (PRA) tools, i.e., 204 semi-structured questionnaire-based individual interviews with fishers, three focus group discussions, and 14 key informants' interviews in three fishing villages in the TH. In contrast, secondary data was set by reviewing published literature and related official documents. Results showed that, due to weak enforcement with inadequate surveillance and poor implementation of the legal framework, there was a high non-compliance with fishing laws, rules, and policies. Destructive and prohibited fishing gears, e.g., the use of small mesh fine nylon nets (current jal), purse seine net (ber jal), and the harvesting during ban period-illicit catch were widespread in the study areas. In addition, catching undersized fish, fishing at the restricted areas (sanctuary area), and fishing during spawning seasons occur often. There is a crying need for a comprehensive legal and policy framework to contextualize the local context, ensure the proper implementation of the fishing laws and regulations, increase the managerial inefficiency of enforcing agencies, ensure livelihood support during the fishing ban, and afford good alternative income options are still significant issues for good governance in the Tanguar Haor ECA. Findings might help to identify the gaps and misunderstanding of the existing legal practice while submitting urgent attention to the need for drawing a comprehensive legal and policy framework (contextually modified according to the local context), taking initiatives and acting synchronously for proper implementation, and calling transdisciplinary collaboration and cooperation among the agencies that may ensure the non-conflicting use of the natural resources of the TH that can be also helpful for the better conservation of this Ramsar wetland.Peer reviewe

Effect of Metarhizium anisopliae (MetA1) on growth enhancement and antioxidative defense mechanism against Rhizoctonia root rot in okra

Rhizoctonia solani is an important necrotrophic pathogenic fungus that causes okra root disease and results in severe yield reduction. Many biocontrol agents are being studied with the intent of improving plant growth and defense systems and reducing crop loss by preventing fungal infections. Recently, a member of the Hypocrealean family, Metarhizium anisopliae, has been reported for insect pathogenicity, endophytism, plant growth promotion, and antifungal potentialities. This research investigated the role of M. anisopliae (MetA1) in growth promotion and root disease suppression in okra. The antagonism against R. solani and the plant growth promotion traits of MetA1 were tested in vitro. The effects of endophytic MetA1 on promoting plant growth and disease suppression were assessed in planta. Dual culture and cell-free culture filtrate assays showed antagonistic activity against R. solani by MetA1. Some plant growth promotion traits, such as phosphate solubilization and catalase activity were also exhibited by MetA1. Seed primed with MetA1 increased the shoot, root, leaves, chlorophyll content, and biomass content compared to control okra plants. The plants challenged with R. solani showed the highest hydrogen peroxide (H2O2) and lipid peroxidation (MDA) contents in the leaves of okra. Whereas MetA1 applied plants showed a reduction of H2O2 and MDA by 5.21 and 14.96%, respectively, under pathogen-inoculated conditions by increasing antioxidant enzyme activities, including catalase (CAT), peroxidase (POD), glutathione S-transferase (GST), and ascorbate peroxidase (APX), by 30.11, 10.19, 5.62, and 5.06%, respectively. Moreover, MetA1 increased soluble sugars, carbohydrates, proline, and secondary metabolites, viz., phenol and flavonoid contents in okra resulting in a better osmotic adjustment of diseases infecting plants. MetA1 reduced disease incidence by 58.33% at 15 DAI compared to the R. solani inoculated plant. The results revealed that MetA1 improved plant growth, elevated the plant defense system, and suppressed root diseases caused by R. solani. Thus, MetA1 was found to be an effective candidate for the biological control program

Precision of the M10 ELISA.

<p>Precision of the M10 ELISA.</p

Establishment of an indirect ELISA for detection of the novel antifibrotic peptide M10

<div><p>Objective</p><p>M10 is a ten amino acid peptide generated from the intracellular cytoplasmic tail of the hepatocyte growth factor (HGF) receptor c-Met following cleavage by caspase-3. Recently we reported that M10 interacts with Smad2 and demonstrates antifibrotic properties in vitro and in vivo and can be advanced into a novel antifibrotic remedy. The current study was undertaken to develop an immunoassay to measure M10 concentration in biological specimens.</p><p>Experimental design</p><p>An Indirect Enzyme-Linked Immunosorbent Assay (ELISA) for detection of M10 in biological fluids was developed using pharmaceutical grade synthetic M10 as a calibrator and commercially available anti-c-Met C12 antibody.</p><p>Results</p><p>M10 ELISA specifically detected in plasma M10, but not a scrambled peptide, following a single intraperitoneal administration of M10 (1mg/kg) to mice. The detection limit was 9.6 ng/ml, and the measuring limit was between 15 ng/ml and 200 ng/ml. The recovery limits of M10 were between 80% and 120%; intra-assay coefficient of variation was between 5.3% and 6.3%; inter-assay coefficient of variation was between 5.0% and 8.0% over the buffer concentration tested in the range from 15 ng /ml to 250 ng /ml. The peak of M10 concentration following a single intraperitoneal injection (1mg/kg) was achieved within 6 hours and declined to minimal levels by 48 hours. The experimentally obtained half-life for M10 was comparable to the theoretically predicted half-life for M10.</p><p>Conclusions</p><p>We have established a highly sensitive ELISA to detect the antifibrotic peptide M10 in plasma samples, which should prove to be a novel tool to study the pharmacokinetics and efficacy of M10 in the treatment of fibroproliferative disorders.</p></div