PSD3 downregulation confers protection against fatty liver disease

Fatty liver disease (FLD) is a growing health issue with burdening unmet clinical needs. FLD has a genetic component but, despite the common variants already identified, there is still a missing heritability component. Using a candidate gene approach, we identify a locus (rs71519934) at the Pleckstrin and Sec7 domain-containing 3 (PSD3) gene resulting in a leucine to threonine substitution at position 186 of the protein (L186T) that reduces susceptibility to the entire spectrum of FLD in individuals at risk. PSD3 downregulation by short interfering RNA reduces intracellular lipid content in primary human hepatocytes cultured in two and three dimensions, and in human and rodent hepatoma cells. Consistent with this, Psd3 downregulation by antisense oligonucleotides in vivo protects against FLD in mice fed a non-alcoholic steatohepatitis-inducing diet. Thus, translating these results to humans, PSD3 downregulation might be a future therapeutic option for treating FLD. Employing a candidate gene approach, Mancina et al. identify a genetic variant of the Pleckstrin and Sec7 domain-containing 3 (PSD3) gene that reduces susceptibility to fatty liver disease. Functional studies in vitro and in vivo demonstrate that targeting PSD3 protects against fatty liver disease.Peer reviewe

Real Time Anti-<em>Toxoplasma gondii </em>Activity of an Active Fraction of <em>Eurycoma longifolia</em> Root Studied by <em>in Situ</em> Scanning and Transmission Electron Microscopy

The inhibitory effect of active fractions of <em>Eurycoma<strong> </strong>longifolia </em>(<em>E. longifolia</em>)<em> </em>root, namely TAF355 and TAF401, were evaluated against <em>Toxoplasma gondii (T. gondii). </em>In our previous study, we demonstrated that <em>T. gondii </em>was susceptible to TAF355 and TAF401 with IC<sub>50</sub> values of 1.125 µg/mL and 1.375 µg/mL, respectively. Transmission (TEM) and scanning electron microscopy (SEM) observations were used to study the <em>in situ </em>antiparasitic activity at the IC<sub>50</sub> value. Clindamycin was used as positive control. SEM examination revealed cell wall alterations with formation of invaginations followed by completely collapsed cells compared to the normal <em>T. gondii </em>cells in response to the fractions. The main abnormality noted via TEM study was decreased cytoplasmic volume, leaving a state of structural disorganization within the cell cytoplasm and destruction of its organelles as early as 12 h of treatment, which indicated of rapid antiparasitic activity of the <em>E. longifolia</em> fractions. The significant antiparasitic activity shown by the TAF355 and TAF401 active fractions of <em>E. longifolia </em>suggests their potential as new anti-<em>T. gondii </em>agent candidates.<em> </em&gt

Structural and electrical performance of Ce0.33Ca0.34Ti0.33O1.66 based solid oxide fuel cell electrolyte fabricated using ball milling techniques

In the present work, calcium and titanium are doped with ceria in an effort to create a unique, effective electrolyte for solid oxide fuel cell applications (Ce0.33Ca0.34Ti0.33O1.66 (CCT-33)). CCT-33 is prepared through solid state method through ball milling and calcination. XRD analysis of the CCT-33 revealed cubic fluorite crystal structure favorable for oxygen ion conduction. Electron microscopy analysis of the sintered pellets revealed good compactness of the phase pure powders and crystallinity. The conductivity studied using impedance analysis is 10−3–10−4 S cm−1 in the temperature region of 500–630 °C. At 700 °C the highest value of 2.86 × 10−2 S cm−1 in considered temperature region was obtained. The present work introduces innovative, modified material for Intermediate Temperature Solid Oxide Fuel Cell (IT-SOFCs)

Human Multilineage 3D Spheroids as a Model of Liver Steatosis and Fibrosis

Non-alcoholic fatty liver disease (NAFLD) is the most common liver disorder in western countries. Despite the high prevalence of NAFLD, the underlying biology of the disease progression is not clear, and there are no approved drugs to treat non-alcoholic steatohepatitis (NASH), the most advanced form of the disease. Thus, there is an urgent need for developing advanced in vitro human cellular systems to study disease mechanisms and drug responses. We attempted to create an organoid system genetically predisposed to NAFLD and to induce steatosis and fibrosis in it by adding free fatty acids. We used multilineage 3D spheroids composed by hepatocytes (HepG2) and hepatic stellate cells (LX-2) with a physiological ratio (24:1). HepG2 and LX-2 cells are homozygotes for the PNPLA3 I148M sequence variant, the strongest genetic determinant of NAFLD. We demonstrate that hepatic stellate cells facilitate the compactness of 3D spheroids. Then, we show that the spheroids develop accumulations of fat and collagen upon exposure to free fatty acids. Finally, this accumulation was rescued by incubating spheroids with liraglutide or elafibranor, drugs that are in clinical trials for the treatment of NASH. In conclusion, we have established a simple, easy to handle, in vitro model of genetically induced NAFLD consisting of multilineage 3D spheroids. This tool may be used to understand molecular mechanisms involved in the early stages of fibrogenesis induced by lipid accumulation. Moreover, it may be used to identify new compounds to treat NASH using high-throughput drug screening

MicroRNAs: biogenesis, roles for carcinogenesis and as potential biomarkers for cancer diagnosis and prognosis

MicroRNAs (miRNAs) are short non-coding RNAs of 20-24 nucleotides that play important roles in carcinogenesis. Accordingly, miRNAs control numerous cancer-relevant biological events such as cell proliferation, cell cycle control, metabolism and apoptosis. In this review, we summarize the current knowledge and concepts concerning the biogenesis of miRNAs, miRNA roles in cancer and their potential as biomarkers for cancer diagnosis and prognosis including the regulation of key cancer-related pathways, such as cell cycle control and miRNA dysregulation. Moreover, microRNA molecules are already receiving the attention of world researchers as therapeutic targets and agents. Therefore, in-depth knowledge of microRNAs has the potential not only to identify their roles in cancer, but also to exploit them as potential biomarkers for cancer diagnosis and identify therapeutic targets for new drug discovery

Genoprotection and cytotoxicity of cassia surattensis seed extract on vero cell evaluated by comet and cytotoxicity assays

Cassia (C.) surattensis Burm. f. (Leguminosae) is a flowering plant that has been traditionally used in many countries for food and medicinal purposes. The present study was conducted to determine in vitro cytotoxicity and anti-genotoxic activity of the seed extract of C. surattensis. The cytotoxic and anti-genotoxic activities were evaluated against Vero cell line using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and comet assays. In vitro cytotoxicity evaluation demonstrated that seed extract of C. surattensis was non-cytotoxic against the Vero cells. The 50 % cytotoxic concentration (CC50) value from MTT cytotoxicity assay was 332.87 µg/ml. The seed extract of C. surattensis exhibited strong inhibitory effects against H2O2-mediated DNA damage in Vero cells as observed in Comet assay. The presence of antioxidant activity in the seed extract was associated with the antigenotoxic property of C. surattensis. In summary, the present study demonstrated an efficient antigenotoxic property of C. surattensis, which is significant for future investigation for human applications

GREEN SYNTHESIS AND CHARACTERIZATIONS OF SILVER NANOPARTICLES USING FRESH LEAF EXTRACT OF MORINDA CITRIFOLIA AND ITS ANTI-MICROBIAL ACTIVITY STUDIES

Objective: To develop a rapid method of synthesis of silver nanoparticles (Ag NPs) using the fresh aqueous leaf extract of Morinda Citrifolia from 0.1 M AgNO3 solution, to characterize the resulting Ag NPs and also compare their antimicrobial activity with those of standard drugs against human pathogenic bacteria.Methods: 25 ml of the aqueous extract was added to 25 ml of 0.1 M aqueous AgNO3 at room temperature. The mixture was stirred continuously for 5-10 minutes. The reduction was completed with the appearance of brownish-black colored dispersion. The resulting nanoparticles were characterized using UV-Vis absorption spectra and Particle size analysis (DLS method). Further the Antimicrobial activity was compared with the drugs against S. aureus and P. aureginosa strains using the disk diffusion method.Results: The formation of Silver nanoparticles was confirmed with the help of UV-Vis absorption spectra at ≈ 425 nm and particle size as approximately 100 nm using Particle size analysis (DLS method). The anti-microbial activity of the Ag NPs so synthesized was studied against human pathogens in wound infections such as S. aureus and P. aureginosa strains. The inhibitory activity for Ag NPs was compared with those of known drugs such as tetracycline, Ceftazidime and Amikacin at 30 mcg. The inhibitory activity of the Synthesized Ag NPs was found pronounced against P. aureginosa strains.Conclusion: A rapid method of synthesizing Ag NPs has been developed by using the fresh leaf extract of Morinda Citrifolia and it was found that the extract is a potential reducing agent to produce stable Ag NPs. The research provides a new input to the development of anti-microbial agent.Â

Impact of passing mesenchymal stem cells through smaller bore size needles for subsequent use in patients for clinical or cosmetic indications

<p>Abstract</p> <p>Background</p> <p>Numerous preclinical and clinical studies have investigated the regenerative potential and the trophic support of mesenchymal stem cells (MSCs) following their injection into a target organ. Clinicians favor the use of smallest bore needles possible for delivering MSCs into vascular organs like heart, liver and spleen. There has been a concern that small needle bore sizes may be detrimental to the health of these cells and reduce the survival and plasticity of MSCs.</p> <p>Methods</p> <p>In this report, we aimed to investigate the smallest possible bore size needle which would support the safe delivery of MSCs into various tissues for different clinical or cosmetic applications. To accomplish this we injected cells via needle sizes 24, 25 and 26 G attached to 1 ml syringe in the laboratory and collected the cells aseptically. Control cells were ejected via 1 ml syringe without any needle. Thereafter, the needle ejected cells were cultured and characterized for their morphology, attachment, viability, phenotypic expression, differentiation potential, cryopreservation and <it>in vivo</it> migration abilities. In the second phase of the study, cells were injected via 26 G needle attached to 1 ml syringe for 10 times.</p> <p>Results</p> <p>Similar phenotypic and functional characteristics were observed between ejected and control group of cells. MSCs maintained their cellular and functional properties after single and multiple injections.</p> <p>Conclusions</p> <p>This study proves that 26 G bore size needles can be safely used to inject MSCs for clinical/therapeutics purposes.</p