Thy-1 plays a pathogenic role and is a potential biomarker for skin fibrosis in scleroderma.

Thy-1 (CD90) is a well-known marker of fibroblasts implicated in organ fibrosis, but its contribution to skin fibrosis remains unknown. We examined Thy-1 expression in scleroderma skin and its potential role as a biomarker and pathogenic factor in animal models of skin fibrosis. Skin from patients with systemic sclerosis demonstrated markedly elevated Thy-1 expression compared with controls, colocalized with fibroblast activator protein in the deep dermis, and correlated with the severity of skin involvement (modified Rodnan skin score). Serial imaging of skin from Thy-1 yellow fluorescent protein reporter mice by IVIS showed an increase in Thy-1 expression that correlated with onset and progression of fibrosis. In contrast to lung fibrosis, Thy-1-KO mice had attenuated skin fibrosis in both bleomycin and tight skin-1 murine models. Moreover, Thy-1 regulated key pathogenic pathways involved in fibrosis, including inflammation, myofibroblast differentiation, apoptosis, and multiple additional canonical fibrotic pathways. Therefore, although Thy-1 deficiency leads to exacerbated lung fibrosis, in skin it is protective. Moreover, Thy-1 may serve as a longitudinal marker to assess skin fibrosis.

MicroRNA-34a-5p Promotes Joint Destruction During Osteoarthritis.

OBJECTIVE: MicroRNA-34a-5p (miR-34a-5p) expression is elevated in the synovial fluid of patients with late-stage knee osteoarthritis (OA); however, its exact role and therapeutic potential in OA remain to be fully elucidated. This study was undertaken to examine the role of miR-34a-5p in OA pathogenesis. METHODS: Expression of miR-34a-5p was determined in joint tissues and human plasma (n = 71). Experiments using miR-34a-5p mimic or antisense oligonucleotide (ASO) treatment were performed in human OA chondrocytes, fibroblast-like synoviocytes (FLS) (n = 7-9), and mouse OA models, including destabilization of the medial meniscus (DMM; n = 22) and the accelerated, more severe model of mice fed a high-fat diet and subjected to DMM (n = 11). Wild-type (WT) mice (n = 9) and miR-34a-knockout (KO) mice (n = 11) were subjected to DMM. Results were expressed as the mean ± SEM and analyzed by t-test or analysis of variance, with appropriate post hoc tests. P values less than 0.05 were considered significant. RNA sequencing was performed on WT and KO mouse chondrocytes. RESULTS: Expression of miR-34a-5p was significantly increased in the plasma, cartilage, and synovium of patients with late-stage OA and in the cartilage and synovium of mice subjected to DMM. Plasma miR-34a-5p expression was significantly increased in obese patients with late-stage OA, and in the plasma and knee joints of mice fed a high-fat diet. In human OA chondrocytes and FLS, miR-34a-5p mimic increased key OA pathology markers, while miR-34a-5p ASO improved cellular gene expression. Intraarticular miR-34a-5p mimic injection induced an OA-like phenotype. Conversely, miR-34a-5p ASO injection imparted cartilage-protective effects in the DMM and high-fat diet/DMM models. The miR-34a-KO mice exhibited protection against DMM-induced cartilage damage. RNA sequencing of WT and KO chondrocytes revealed a putative miR-34a-5p signaling network. CONCLUSION: Our findings provide comprehensive evidence of the role and therapeutic potential of miR-34a-5p in OA.

Effect of autotaxin inhibition in a surgically-induced mouse model of osteoarthritis.

Objective: Osteoarthritis (OA) is a degenerative joint disease with no approved disease modifying therapy. The enzyme autotaxin (ATX) converts lysophoshatidylcholine analogues to lysophosphatidic acid. Systemic inhibition of ATX reduces pain in animal models of OA; however, OA disease-modifying effects associated with ATX inhibition remain unknown. Here, we sought to determine whether local (knee joint) injection of an ATX inhibitor attenuates surgically-induced OA in mice. Methods: ATX expression was evaluated in human knee OA cartilage. Ten-week-old mice were subjected to surgically-induced OA. ATX inhibitor (PF-8380, 2.5ng/joint) was injected intra-articularly either at three and five weeks post-surgery or at two, four, six and eight weeks post-surgery and knee joints were evaluated by histopathology and immunohistochemistry to study the expression of catabolic and cell death markers. mRNA sequencing of human OA chondrocytes treated with/without ATX inhibitor was performed to identify differentially expressed transcripts, followed by pathway enrichment analysis. Results: ATX expression was elevated in severely degenerated cartilage compared to less degenerated human OA cartilage. In surgically-induced OA mice, intra-articular injection of ATX inhibitor at three and five weeks post-surgery partially protected knee joints from cartilage degeneration. Interestingly, earlier and more frequent ATX inhibitor injections did not confer significant protection. Immunohistochemical analysis showed decreased expression of catabolic and apoptotic markers with two ATX inhibitor injections. mRNA sequencing followed by pathway analysis identified pathways related to cholesterol analogue metabolism and cell-cycle that could be modulated by ATX inhibition in human OA chondrocytes. Conclusion: Local delivery of ATX inhibitor partially attenuates surgically-induced OA in mice.

Production of novel rhamnolipids via biodegradation of waste cooking oil using Pseudomonas aeruginosa MTCC7815.

In this paper, Pseudomonas aeruginosa MTCC7815, a biosurfactant producing strain was studied for its ability to utilize waste cooking oil (WCO) as a sole carbon source for the production of biosurfactant. Culture conditions were optimized based on surface tension reduction and biomass concentration. The obtained biosurfactant was characterized using 1H NMR, FTIR, LC-MS, and MALDI-TOF techniques. The chemical properties of the produced biosurfactant were estimated by assessing the critical micelle concentration (CMC), emulsification index (E24) and oil displacement test. The optimal culture conditions were found to be similar to the natural domestic sewage such as basic pH value of 10, temperature of 25 °C and a very high WCO concentration of 40 gL-1 (C/N ratio of 40/1). The biosurfactant yield was found to be significant as 11 ± 0.2 gL-1 upon utilizing about 90% of WCO within 5 days of incubation. The biosurfactant produced was found to be a mixture of mono- and di-rhamnolipid in nature and comprised excellent surface active properties i.e. an extremely low CMC of 8.8 ± 0.3 mgL-1, E24 of 62.5 ± 0.3% and surface tension reduction up to 26.2 ± 0.5 mNm-1. These results suggest the suitability of Pseudomonas aeruginosa for the biosurfactant production at commercial scale along with waste remediation in an economic way.

Isolation and characterization of biosurfactant producing and oil degrading Bacillus subtilis MG495086 from formation water of Assam oil reservoir and its suitability for enhanced oil recovery.

The strains isolated from the formation water were characterized and screened considering their crude oil degradation capability and biosurfactant production ability. The growth kinetics study of isolated Bacillus subtilis MG495086 was carried out by varying growth parameters i.e. carbon source, temperature, pH and salinity. The biosurfactant production was optimized adopting RSM-CCD considering carbon source (1-5%), pH (3-11) and temperature (25-65 °C) as matrix parameters. The optimum biosurfactant production (6.3 ±â€¯0.1 g/L) and the minimum surface tension 29.85 mN/m were obtained after 96 h of incubation under optimal conditions i.e. 3.8% (v/v) of light-paraffin oil as sole carbon source at 62.4 °C and pH 7.7 with the maximum oil degradation capability of 91.3 ±â€¯5%. Critical micelle concentration value of crude biosurfactant was found to be 40 mg/L with high emulsification activity of 72.45 ±â€¯0.85%. The produced biosurfactant was identified as lipopeptide (Surfactin) and characterized using various analytical techniques to establish its suitability for microbial enhanced oil recovery.

Inhibition of Wnt/ß-catenin signaling ameliorates osteoarthritis in a murine model of experimental osteoarthritis.

Osteoarthritis (OA) is a degenerative joint disease involving both cartilage and synovium. The canonical Wnt/ß-catenin pathway, which is activated in OA, is emerging as an important regulator of tissue repair and fibrosis. This study seeks to examine Wnt pathway effects on synovial fibroblasts and articular chondrocytes as well as the therapeutic effects of Wnt inhibition on OA disease severity. Mice underwent destabilization of the medial meniscus surgery and were treated by intra-articular injection with XAV-939, a small-molecule inhibitor of Wnt/ß-catenin signaling. Wnt/ß-catenin signaling was highly activated in murine synovial fibroblasts as well as in OA-derived human synovial fibroblasts. XAV-939 ameliorated OA severity associated with reduced cartilage degeneration and synovitis in vivo. Wnt inhibition using mechanistically distinct small-molecule inhibitors, XAV-939 and C113, attenuated the proliferation and type I collagen synthesis in synovial fibroblasts in vitro but did not affect human OA-derived chondrocyte proliferation. However, Wnt modulation increased COL2A1 and PRG4 transcripts, which are downregulated in chondrocytes in OA. In conclusion, therapeutic Wnt inhibition reduced disease severity in a model of traumatic OA via promoting anticatabolic effects on chondrocytes and antifibrotic effects on synovial fibroblasts and may be a promising class of drugs for the treatment of OA.

High-fat diet-induced acceleration of osteoarthritis is associated with a distinct and sustained plasma metabolite signature.

Metabolic changes induced by high fat diet (HFD) that contribute to osteoarthritis (OA) are poorly understood. We investigated longitudinal changes to metabolites and their contribution to OA pathogenesis in response to HFD. HFD-fed mice exhibited acceleration of spontaneous age-related and surgically-induced OA compared to lean diet (LD)-fed mice. Using metabolomics, we identified that HFD-fed mice exhibited a distinct and sustained plasma metabolite signature rich in phosphatidylcholines (PC) and lysophosphatidylcholines (lysoPCs), even after resumption of normal chow diet. Using receiver operator curve analysis and prediction modelling, we showed that the concentration of these identified metabolites could efficiently predict the type of diet and OA risk with an accuracy of 93%. Further, longitudinal evaluation of knee joints of HFD- compared to LD- fed mice showed a greater percentage of leptin-positive chondrocytes. Mechanistic data showed that leptin-treated human OA chondrocytes exhibited enhanced production of lysoPCs and expression of autotaxin and catabolic MMP-13. Leptin-induced increased MMP13 expression was reversed by autotaxin inhibition. Together, this study is the first to describe a distinct and sustained HFD-induced metabolite signature. This study suggests that in addition to increased weight, identified metabolites and local leptin-signaling may also contribute in part, towards the accelerated OA-phenotype observed in HFD mice.

Deciphering the mechanistic insight into the stoichiometric ratio dependent behavior of Cu(II) on BSA fibrillation.

Various metal ions are recently implicated in protein aggregates and are associated with numerous neurodegenerative diseases. In the present work, we have scrutinized the effect of stoichiometric variation of Cu(II) on BSA fibrillation at physiological pH 7.4 through Thioflavin-T dye binding study, residual protein concentration, Fourier Transform Infrared spectroscopy (FTIR), Dynamic Light Scattering (DLS), Atomic Force Microscopy (AFM) and Isothermal Titration Calorimetry (ITC). Fibrillation kinetics was studied through ThT fluorescence, which illustrated dependency on stoichiometric ratio of Cu(II). The ThT intensity data were fitted to a single exponential expression to determine the aggregation rate, k, which revealed a slight lower k value for 1:1 ratio of BSA-Cu(II) reaction as compared to BSA alone whereas k value for 1:2 ratio of BSA-Cu(II) reaction was higher. Higher equilibrium residual concentration in case of 1:1 ratio of BSA-Cu(II) agreed to the lower aggregation rate. IR spectroscopy revealed the presence of increased ß-sheet proportion to the detriment of α-helix conformation with increasing concentration of Cu(II) and illustrated maximum ß-sheet proportion in 1:2 ratio of BSA-Cu(II). These results were combined with scattering results that showed the higher average hydrodynamic radius (Z-average value) of aggregates in 1:2 ratio of BSA-Cu(II) with respect to BSA and 1:1 ratio of BSA-Cu(II). AFM analysis confirmed the fibril formation. ITC analysis has shown the presence of two binding sites with many fold difference in binding affinity at prescribed in vitro conditions. An N-terminal binding site with many fold higher binding affinity was found as the first Cu(II) binding site. The free thiol group of the cysteine residue positioned at 34 (cys-34) in BSA was covalently capped and this modified BSA also showed the presence of two binding sites, which declined the cys-34 site as the second Cu(II) bind site. Therefore Zn(II) binding site was predicted as the second Cu(II) binding site. The binding of Cu(II) at N-terminal binding site marks a square planner geometry; while that at zinc binding site marks a rigid coordination geometry. Conclusively, analyzed data has concluded that the variation in Cu(II) efficacy in the different stoichiometric ratio results because of two different coordination types in different stoichiometric ratios.

Identification of microRNA-181a-5p and microRNA-4454 as mediators of facet cartilage degeneration.

Osteoarthritis (OA) of spine (facet joints [FJs]) is one of the major causes of severe low back pain and disability worldwide. The degeneration of facet cartilage is a hallmark of FJ OA. However, endogenous mechanisms that initiate degeneration of facet cartilage are unknown, and there are no disease-modifying therapies to stop FJ OA. In this study, we have identified microRNAs (small noncoding RNAs) as mediators of FJ cartilage degeneration. We first established a cohort of patients with varying degrees of facet cartilage degeneration (control group: normal or mild facet cartilage degeneration; FJ OA group: moderate to severe facet cartilage degeneration) and then screened 2,100 miRNAs and identified 2 miRNAs (miR-181a-5p and miR-4454) that were significantly elevated in FJ OA cartilage compared with control facet cartilage. We further explored their role, function, and signaling mechanisms using computational, in vitro functional, and in vivo studies. We specifically indicate that miR-181a-5p and miR-4454 are involved in promoting inflammatory, catabolic, and cell death activity in FJ chondrocytes. This is the first report to our knowledge that identifies miR-181a-5p and miR-4454 as mediators of cartilage degeneration in FJs and potential therapeutic targets for stopping cartilage degeneration.

Physiological Interactions of Nanoparticles in Energy Metabolism, Immune Function and Their Biosafety: A Review.

Nanoparticles owing to their unique physico-chemical properties have found its application in various biological processes, including metabolic pathways taking place within the body. This review tried to focus the involvement of nanoparticles in metabolic pathways and its influence in the energy metabolism, a fundamental criteria for the survival and physiological activity of living beings. The human body utilizes energy derived from food resources through a series of biochemical reactions involving several enzymes, co-factors (metals, non-metals, vitamins etc.) through the metabolic pathways (glycolysis, tri carboxylic acid cycle, oxidative phosphorylation, electron transport chain, etc.) in cellular system. Energy metabolism is also involved in the immune networking of the body for self defence and against pathophysiology. The immune system comprises of different cells and tissues, bioactive molecules for self defence and to fight against diseases. In the recent times, it has been reported through in vivo and in vitro studies that nanoparticles have direct influence on body's immune functions, and can modulate immunity by either suppressing or enhancing it. A comprehensive overview of nanoparticles and its involvement in immune function of the body in normal and pathophysiological conditions has been discussed. Considering these perspectives on nanoparticle interaction another important area which has been highlighted is the biosafety issues which are necessary before therapeutic applications. It is expected that development of physiologically compatible nanoparticles controlling energy metabolic processes, immune functions may show new dimension in the pathophysiology linked with energy and immunity.

Anti arthritic and anti inflammatory activity of a cytotoxic protein NN-32 from Indian spectacle cobra (Naja naja) venom in male albino rats.

The anti arthritic and anti inflammatory activity of NN-32, a cytotoxic protein from Indian spectacle cobra snake (Naja naja) venom has been studied in Freund's complete adjuvant (FCA) induced arthritis and carrageenan induced anti inflammatory model. NN-32 treatment showed significant decrease in physical and urinary parameters, serum enzymes, serum cytokines levels as compared to arthritic control group of rats. NN-32 treatment recovered carrageenan induced inflammation as compared to control group of rats. The findings showed that the cytotoxic protein NN-32 shares anti arthritic and anti inflammatory activity and thus NN-32 may target complex pathophysiological processes like cancer- arthritis-inflammation.

Anti-rheumatoid and anti-oxidant activity of homeopathic Guaiacum officinale in an animal model.

BACKGROUND: Homeopathy is a popular form of complementary and alternative medicine. Guaiacum extract is said to be useful for pain and inflammation, but there appears to be no scientific evidence to support this. AIMS: The aim of the present study was to evaluate the anti-rheumatic and anti-oxidant activity of homeopathic preparations of Guaiacum officinale (Gua) on experimental animal model. DESIGN: Rheumatoid arthritis (RA) was induced in male albino rats by Freund's complete adjuvant (FCA) at a dose of (0.25 mg heat killed Mycobacterium tuberculosis/ml of emulsion). Gua mother tincture (MT) (prepared from the latex part of the plant) (MT), Gua 30cc and 200cc were purchased commercially from King Company, Kolkata, India. Male albino Wistar rats (130 ± 10 g) were divided into 6 groups: Sham control; Arthritis control; Standard treatment indomethacin (0.25 mg 100 g(-1) p.o. × 5 alternative days), Gua MT (1 ml kg(-1) p.o. × 5 days) treated; Gua (30c 1 ml kg(-1) p.o. × 5 days) treated; Gua (200c; 1 ml kg(-1) p.o. × 5 days) treated. Anti-rheumatic activity was examined through physical, urinary, serum parameters. All the results were expressed in terms of mean ± SEM (statistical error of mean n = 6) at each dose level. The level of significance was determined through one-way analysis of variance (ANOVA), p < 0.05 was considered significant. RESULTS: It was observed that body weight, ankle and knee diameter, urinary parameters (hydroxyproline (OH-P), glucosamine, calcium (Ca(2)(+)), creatinine (CRE), phosphate (PO4(3)(-))), serum ACP (acid phosphatase)/ALP (alkaline phosphatase)/Ca(2+)/CRE/PO4(3-)/gamma-glutamyl transferase (GGT)/Lipid peroxidation (LPO)/Glutathione (GSH)/Superoxide dismutase (SOD)/Catalase, serum GGT, serum interleukins like IL-1ß/CINC-1/PGE2/TNF-α/IL-6, IL-12/IL-4/IL-6 levels were significantly affected. After treatment with Guaiacum in all 3 regimes was associated with normalization of these parameters compared to control group. CONCLUSION: These findings suggest that homeopathic G. officinale possesses anti-rheumatic and anti-oxidant activity in experimental animal and these activities may be more significant in higher potencies.

Physiologically important metal nanoparticles and their toxicity.

Nanotechnology has been setting benchmarks for the last two decades, but the origins of this technology reach back to ancient history. Today, nanoparticles of both metallic and non-metallic origin are under research and development for applications in various fields of biology/therapeutics. Physiologically important metals are of concern because they are compatible with the human system in terms of absorption, assimilation, excretion, and side effects. There are several physiologically inorganic metals that are present in the human body with a wide range of biological activities. Some of these metals are magnesium, chromium, manganese, iron, cobalt, copper, zinc, selenium and molybdenum. These metals are synthesized in the form of nanoparticles by different physical and chemical methods. Physiologically important nanoparticles are currently under investigation for their bio-medical applications as well as for therapeutics. Along with the applicative aspects of nanoparticles, another domain that is of great concern is the risk assessment of these nanoparticles to avoid unnecessary hazards. It has been seen that these nanoparticles have been shown to possess toxicity in biological systems. Conventional physical and chemical methods of metal nanoparticle synthesis may be one possible reason for nanoparticle toxicity that can be overcome by synthesis of nanoparticles from biological sources. This review is an attempt to establish metal nanoparticles of physiological importance to be the best candidates for future nanotechnological tools and medicines, owing to the acceptability and safety in the human body. This can only be successful if these particles are synthesized with a better biocompatibility and low or no toxicity.

In vivo interaction of gold nanoparticles after acute and chronic exposures in experimental animal models.

With emerging use of gold nanopaticles (GNP) in biomedical science now concern lies upon the fact that how this nonanparticles interact with biological systems both in vivo and in vitro. In this study effects of GNP (50 nm) were investigated in animal models after acute and chronic exposure. For acute studies GNP was administered intravenously at three doses and urine and blood samples were collected for urinary and haematological analysis at regular time intervals. For chronic studies GNP was administered intra-peritoneally at two dose levels and urine, blood, serum and tissue samples were collected for urinary, haematological, serum biochemical and histo-pathological analysis at regular intervals. Acute exposure revealed significant increase in WBC count at all the three dose levels and significant dose-dependent increase in RBC count and Hb%. Chronic exposure at 2 mg/kg dose level showed high toxicity. Significant changes in physical, morphological, WBC count and Hb% were observed after chronic exposure for multiple days. Histo-pathological studies indicated detrimental tissue histological changes in chronic animal models. Therefore, the above studies indicate that both acute and chronic GNP exposure exhibits potential physiological changes within animal system.

Nanoparticle-conjugated animal venom-toxins and their possible therapeutic potential.

Nano-medical approaches to develop drugs have attracted much attention in different arenas to design nanoparticle conjugates for better efficacy of the potential bio-molecules. A group of promising candidates of this category would be venom-toxins of animal origin of potential medicinal value. Traditional systems of medicine as well as folklores mention the use of venom-toxins for the treatment of various diseases. Research has led to scientific validation of medicinal applications of venoms-toxins and many active constituents derived from venoms-toxins are already in clinical use or under clinical trial. Nanomedicine is an emerging field of medicine where nanotechnology is used to develop molecules of nano-scale dimension, so that these molecules can be taken up by the cells more easily and have better efficacy, as compared to large molecules that may tend to get eliminated. This review will focus on some of the potential venoms and toxins along with nanoparticle conjugated venom-toxins of snakes, amphibians, scorpions and bees, etc., for possible therapeutic clues against emerging diseases.

Hepatoprotective activity of the edible snail (Bellamia bengalensis) flesh extract in carbon tetrachloride induced hepatotoxicity in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: In the folk-traditional medicine, snails were used to purify blood, boost immune system, prevent conjunctivitis and to treat liver problems. OBJECTIVES: To evaluate the hepatoprotective activity of the edible snail (Bellamia bengalensis) flesh extract in male Wistar rats treated with carbon tetrachloride as an hepatotoxicant. MATERIALS AND METHODS: Live adult Bellamia bengalensis was collected commercially from the Kolkata market. Aqueous flesh extract (BBE) was prepared in 0.9% saline and expressed in terms of wet weight basis. The aqueous flesh extract was administered orally (1, 2 g kg(-1)day(-1)) to male rats for 12 days. Liv52 was used as positive control. 24h after administration of extract, the rats were given a single oral dose of CCl(4) (1.25 ml kg(-1)), except vehicle control rats. After 24h of CCl(4) administration, all the animals were sacrificed to collect the blood and liver tissue. RESULTS: BBE (1 and 2 g kg(-1)day(-1), p.o.×12 days) significantly prevented CCl(4) induced elevation of SGOT, SGPT, γGT, ACP, ALP, bilirubin, LDH and CCl(4) induced decrease in total protein, triglyceride level in male Wistar rats. BBE treated rat liver anti-oxidant parameters (LPO, SOD, GSH, CAT, GPx) were significantly antagonized for the pro-oxidant effect of CCl(4). Histopathological studies also supported the protective effect of BBE. CONCLUSION: This study validated the folk and traditional use of snail in liver disorder through CCl(4)-induced rat experimental model.