Search

Background and Objectives Composting is an effective approach for organic waste management, but it also has disadvantages such as greenhouse gases emission (CH4, CO2, and N2O), bad smells, nitrogen loss, and contamination of soil and water resources. Therefore, it is crucial to develop a method to reduce gas emissions and improve the quality of compost at the same time. To overcome these disadvantages, some additives are usually used. The use of additives at the beginning of the composting process to produce valuable compost is known as "cocomposting". Various materials can be incorporated into waste during composting. These materials fall into three categories: organic, mineral, biological, or a combination of these. Some of these added materials serve as bulking agents, primarily affecting the physical structure of the compost (such as aeration). However, most of the time, these substrates also have direct or indirect impacts on other composting factors and can be considered as additives. Additives play a role in enhancing the composting process by reducing leaching and gas emissions, improving aeration, accelerating organic matter breakdown, and enhancing nutrient content and availability in the final product. The research published so far showed that the effect of wood biochar along with leonardite and coal on the quality of compost obtained from animal manure and forest organic matter had not been studied in Iran, and considering the importance of the feasibility of improving the quality of compost, this research was conducted. Methodology In this research, the effect of wood biochar, leonardite and coal on some characteristics and quality indices of the co-compost of animal manure and forest organic matter was studied. The biochar was produced at 400° C from the pyrolysis of mixed pruning plum and pomegranate brunches. Leonardite and coal were also prepared from companies active in this field. The experiment was conducted as a factorial in a completely random basic design with three replications. The first factor of treatment included wood biochar, leonardite and coal at two levels of 2 and 4% by dry weight and the second factor of time included 12 weeks. During the composting process, temperature, pH, EC, total nitrogen concentration, organic carbon content and some quality indices of composting such as humification index (HR), humification ratio (HI), degree of polymerization (DP) and total materials Humic (HS) were measured. Findings According to the results, the treatments were significantly different from the control regarding the temperature of the compost pile, and the coal treatment showed the highest temperature at the level of 2%, and the duration of the thermophilic phase in the leonardite treatment was 2% longer than the other treatments. Among the additives, 2% and 4% leonardite treatments created the highest total nitrogen content in the compost. The additives used in this research did not make a significant difference in compost pH, but coal at the level of 4% caused a significant increase in electrical conductivity (EC). Biochar and coal increased organic carbon concentration in the compost pile. Leonardite treatments of 2% and 4% produced the highest values of humification indices (HS, DP, HI, HR), but they were not significantly different from each other. The biochar used in this research increased the C/N ratio of co-compost. Conclusion In general, leonardite treatments were found to be useful in terms of the indicators of final compost and biochar and coal treatments to accelerate composting in the early stages. But the results of this research showed that it can be used at the end of composting processes in order to enrich and improve the quality of the produced compost. The results of this research showed that considering the costs of raw materials, coal is a suitable treatment to accelerate the production and improve the quality of compost. The results obtained about the effect of biochar obviously cannot be generalized to all biochars and different results may be obtained depending on the type of biomass and pyrolysis conditions. Therefore, it is recommended to use the combination of organic and biological or organic and inorganic additives in future research. Other traits such as indices of microbial contamination and abundance of weed seeds should also be measured and the effect of treatments should be studied. Finally, it is recommended to study the effect of final compost on the characteristics of calcareous and acidic soils. [ABSTRACT FROM AUTHOR]

Introduction Oil contamination affects the biological, physical, and chemical properties of soil. The abundance and diversity of soil microbial communities can significantly be influenced by petroleum hydrocarbons. Soil biological indicators including microbial population and enzyme activity, are highly sensitive to environmental stresses and respond to them quickly. Measuring the microbial population is one of the most common biological indicators which is used to study the quality and health of the soil. Also, measuring the activity of enzymes such as urease is one of the most sensitive indicators of oil-contaminated soils. There are some studies on the effects of oil contamination on microbial population and soil enzyme activity. Most of the studies have tested non-natural and short-term oil pollution and reported the adverse effects of oil hydrocarbons on microbial activities in soil. While the soil sample used in this research had natural and long-term contamination and the microorganisms are compatible with polluted conditions. The aim of this study was to investigate changes in the microbial population and urease activity in the presence of different levels of oil contamination, and how petroleum hydrocarbons can affect them. Petroleum hydrocarbons are toxic and persistent in soil, so it is necessary to study the pattern of changes in soil biological characteristics in effective soil management. Material and Methods In this study, 120 samples of oil-contaminated soils were collected from the oil-rich area of Naft-Shahr (located in the west of Kermanshah province) which had natural and long-term oil pollution. A nested design was used to analysis data in this research. The test factors included locations (4 locations) and 3 different levels of oil pollution: low (L), moderate (M), and high (H). Also, 10 replications were considered in the three levels of oil contamination. The collected soils were analyzed for physico-chemical (pH, EC, 0m, CCE, OC, soil texhure) and biological properties (including urease activity, BR and SIR) using standard methods, and the concentration of oil pollutants was determined by the Soxhlet extractor. To determine the abundance of the culturable microbial population, bacterial counting was performed using nutrient agar (NA) and carbon-free minimal medium (CFMM) supplemented with crude oil as the media. Urease activity was measured by the indophenol blue method and finally, the results of measuring chemical, physical and biological properties were analyzed using principal component analysis (PCA). Results and Discussion The average percentage of oil measured by Soxhlet method was 4.03%, 9.95% and 22.50% respectively for L, M and H levels. The results showed that the microbial population increased with the increase of contamination intensity. The highest microbial population counted in NA clhrre medium was 9.54 x105 CFU/g in H soils and the lowest population was 3.25 x 105 CFU/g in L soils. In the CFMM culture medium, the highest population in H soils was 11.3 x 105 CFU/g and the lowest population in L soils was 11.8 x 104 CFU/g. For both NA and CFMM mediums, location 1 had the highest population and location 4 had the lowest microbial population. Oil contamination of soil samples led to a decrease in urease activity in such a way that the highest enzyme activity in soils was obtained with low contamination (594.90 jrgNHgh) and the lowest activity in heavily contaminated soils (176.11 gNHgh). Also, the lowest urease activity was observed in location 1 and the highest in location 4. Principal components analysis (PCA) was also performed and 71% of the variance of the samples could be explained by the first two components (biochemical component and physical component). The results of this research indicated an increase in the microbial population with an increasing of the intensity of oil pollution. It seems that the results obtained from the studies conducted on man-made pollution and natural pollution have differences in terms of the type of biological responses. Aged, long-term and natural oil pollution has caused the selection of oil-resistant microbial community, and therefore we see their positive response to the presence of oil compounds. Conversely, urease enzyme activity was found to be higher in soils with low pollution. This suggests that microbial activity, while influential, is not the sole determinant of urease activity, and various factors contribute to Soil Enzyme Activity (SEA). The type of petroleum pollutant, the direct effect of petroleum compounds on urease-producing microorganisms, as well as the non-microbial origin of urease in soil can be possible reasons for reducing urease activity in contaminated soils. Conclusion In areas where petroleum pollutants are naturally and long-term present in the soil, some oil-decomposing microbial groups use petroleum hydrocarbons as a source of carbon for their nutrition, so the abundance of oildecomposing communities increases. The results showed an increase in the microbial population with an increase in the intensity of oil pollution. On the other hand, the activity of urease enzyme measured in soils with low pollution was higher because non-microbial factors may affect the activity of this enzyme and the increase in the microbial population is not related to the increase in the population of urease-producing microbes. [ABSTRACT FROM AUTHOR]

Bacteria that colonize plant roots and promote plant growth are referred to as plant growth-promoting rhizobacteria (PGPR). For a long-serving period, the PGPRs have been applied as biofertilizers in crops culture. Recent studies indicated the importance of PGPR for controlling the water deficit. The present study investigates the effects of two different PGPRs on some morphophysiological characteristics in fenugreek under water deficit stress. The first factor was application of four PGPR levels including (1. Sinorhizobium meliloti, 2. Pseudomonas fluorescens, 3. combination of S. meliloti and P. fluorescens and 4. control without bacterial inoculation) and four levels of soil water content including 40%, 60%, 80% and 100% of field capacity (FC) was considered as second factor. The results showed that, leaf area, shoot fresh and dry weight, nitrogen, phosphorus and potassium content, and water use efficacy (WUE) were significantly improved by PGPR inoculation and individual use of PGPR was more effective. Decreasing of soil water content up to 0.40 FC and inoculation of two bacteria led to increase of secondary metabolites such as nicotinic acid and trigonelline. However seed yield was decreased in PGPR treated plants. [ABSTRACT FROM AUTHOR]

Fenugreek (Trigonella foenum-graecum L.) is a valuable medicinal plant, which is widely distributed throughout the world. It has been known that plant growth promoting rhizobacteria (PGPR) have positive effects on the quality and quantity of medicinal plants under different soil water levels. For this reason, a factorial experiment was conducted on the basis of a randomized complete block design (RCBD) to evaluate PGPR effects on the morphophysiological and phytochemical traits of fenugreek under different soil water levels. This study was conducted in two separate experiments: after the six-leaf stage and after the flowering stage. In the experiments, the treatments were plant growth promoting rhizobacteria (PGPR) including the control, Sinorhizobium meliloti, Pseudomonas fluorescens, a combination of S. meliloti and P. fluorescens, and different soil water levels (i.e. 100, 80, 60 and 40% of field capacity (FC) in three replications. The results showed that the highest seed weight per plant was obtained by inoculation with the S. meliloti and P. fluorescens combination at 100% FC after the two developmental stages. The maximum concentrations of nicotinic acid and trigonelline were observed for the combination of S. meliloti and P. fluorescens at the soil water content of 40% FC after the six-leaf stage and for S. meliloti at the soil water content of 40% FC after the flowering stage. The correlation and stepwise regression analyses showed positive effects of PGPR application on the morphophysiological and phytochemical traits of fenugreek plants under different soil water levels.

Introduction: Many studies have been done on the use of aloe vera in wound healing, but fewer studies were done on the influence of this material on the reduction of the alar scar. Therefore, we evaluated the effect of a newly made aloe vera cream on alar wound healing after rhinoplasty., Materials and Methods: This was a randomized, double-arm, parallel-group, double-blind controlled trial and was done from June 2021 to February 2022. External wedge resection was done for all patients. The patients were randomly assigned to receive aloe vera cream (n=31) (intervention group) or Face Doux cream (comparison group) (n = 29). A pharmacist prepared the aloe vera cream. The primary outcome measure was the wound scar status which was assessed by two Questionnaires, including the mean Patient Scar Assessment Questionnaire (PSAQ) and Vancouver Scar Scale (VSS). Randomization and Blinding were done., Results: The mean PSAQ was significantly lower in group A after two weeks (26.9 versus 31.5, P<0.001), after two months (15.7 versus 19.6, P=0.04), and six months follow-up (8.8 versus 11.8, P=0.005). The mean VSS was significantly lower in group A after two weeks (5.6 versus 7.1, P=0.001), after two months (3.5 versus 4.9, P=0.002), and six months (1.2 versus 2.7, P<0.001). Repeated measurement analysis showed that both interventions significantly affected PSAQ and VSS., Conclusion: Although both interventions had a significant effect on PSAQ and VSS, compared to Face Duox, the topical use of Aloe Vera cream significantly reduced scar formation after alar resection, both statistically and clinically.

Helminth parasites modulate the host immune system to ensure a long-lasting asymptomatic form of infection generally, mediated by the secretion of immunomodulatory molecules and one such molecule is a homologue of human host cytokine, Macrophage migratory Inhibitory Factor (hMIF). In this study, we sought to understand the role of homologue of hMIF from the lymphatic filarial parasite, Wuchereria bancrofti (Wba-MIF2), in the immunomodulation of the Streptozotocin (STZ)-induced Type1 Diabetes Mellitus (T1DM) animal model. Full-length recombinant Wba-MIF2 was expressed and found to have both oxidoreductase and tautomerase activities. Wba-MIF2 recombinant protein was treated to STZ induced T1DM animals, and after 5 weeks pro-inflammatory (IL-1, IL-2, IL-6, TNF-α, IFN-γ) and anti-inflammatory (IL-4, IL-10) cytokines and gene expressions were determined in sera samples and spleen respectively. Pro-inflammatory and anti-inflammatory cytokine levels were significantly (p<0.05) up-regulated and down-regulated respectively, in the STZ-T1DM animals, as compared to treated groups. Histopathology showed macrophage infiltration and greater damage of islets of beta cells in the pancreatic tissue of STZ-T1DM animals, than Wba-MIF2 treated STZ-T1DM animals. The present study clearly showed the potential of Wba-MIF2 as an immunomodulatory molecule, which could modulate the host immune system in the STZ-T1DM mice model from a pro-inflammatory to anti-inflammatory milieu., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Glucagon signaling is essential for maintaining normoglycemia in mammals. The arrestin fold superfamily of proteins controls the trafficking, turnover, and signaling of transmembrane receptors as well as other intracellular signaling functions. Further investigation is needed to understand the in vivo functions of the arrestin domain-containing 4 (ARRDC4) protein family member and whether it is involved in mammalian glucose metabolism. Here, we show that mice with a global deletion of the ARRDC4 protein have impaired glucagon responses and gluconeogenesis at a systemic and molecular level. Mice lacking ARRDC4 exhibited lower glucose levels after fasting and could not suppress gluconeogenesis at the refed state. We also show that ARRDC4 coimmunoprecipitates with the glucagon receptor, and ARRDC4 expression is suppressed by insulin. These results define ARRDC4 as a critical regulator of glucagon signaling and glucose homeostasis and reveal a novel intersection of insulin and glucagon pathways in the liver., Competing Interests: Conflict of interest R. T. L. is a cofounder, scientific advisory board member, and private equity holder of Elevian, Inc. R. T. L. is a member of the scientific advisory board of Revidia Therapeutics, Inc, and the laboratory receives research support from BlueRock Therapeutics. J. K. K. is a scientific advisor of Elevian, Inc. All other authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Introduction: In order to improve the efficacy of requesting knee radiography and reduce unnecessary radiation exposure, some clinical decision rules have been proposed for the assessment of knee injuries. Among them, the Ottawa Knee Rule (OKR) was considered as one of the best guidelines with several validation studies. Therefore, in this meta-analysis, we aimed to investigate the accuracy of OKR for diagnosis of fracture in patients presenting with knee trauma., Methods: A systematic search was conducted in PubMed, Web of Science, Scopus, Google Scholar, and EBSCO from inception to September 2022. Quality assessment of the included studies was performed using QUADAS-2 tool. Diagnostic accuracy parameters were analyzed using random-effects model. Statistical analysis was performed using Meta-Disc and Stata softwares., Results: The meta-analysis of the 18 included studies (6702 patients) showed that the pooled sensitivity and specificity of OKR for diagnosis of fractures were 0.98 (95% CI: 0.96-0.99) and 0.43 (95% CI: 0.42-0.45), respectively. The pooled positive likelihood ratio (PLR) and negative likelihood ratio (NLR) were 1.56 (95% CI: 1.39-1.75) and 0.12 (95% CI: 0.05-0.26), respectively. The area under curve (AUC) of the hierarchical summary receiver operating characteristic (HSROC) curve was 0.54., Conclusion: This meta-analysis indicates that OKR has a high diagnostic performance for diagnosis of fracture, with a pooled sensitivity of 98% and a pooled specificity of 43%. These results propose potential effects of OKR on reduction of unnecessary radiography, time spent in emergency departments, and direct and indirect costs, which should be confirmed using high-quality studies in the future., Competing Interests: None.

Until now, a few studies have been conducted on the destructive effects of TiO 2 NPs in living organisms, and studies on the toxicity of TiO 2 NPs are still in the beginning phases. Because of the widespread use of TiO 2 NPs in all areas of human life, it is essential to study their profound and fundamental toxic effects on each organ and body cell. Herein, we evaluate the effect of exposure to TiO 2 NPs on in vitro models derived from the rat bone marrow and adipose tissues. Exposure to TiO 2 NPs at 100 and 200 μg/ml exhibited cytotoxicity for the rat bone marrow mesenchymal stem cells (rBMSCs) and rat adipose mesenchymal stem cells (rATSC), respectively. Additionally, reduced rBMSCs and rATSCs frequencies in the S phase of the cell cycle. Moreover, TiO 2 NPs enhanced the activity of cellular senescence-associated β-galactosidase in both model cells. Significantly higher relative expression of aging-related genes P53 and NF-kB (p < 0.05) and lower expression levels of anti-aging-related genes Nanog and SIRT1 were found in the treated cells (p < 0.05). Colony-forming and DAPI staining showed the reduction of cell growth and DNA damage in both rBMSCs and rATSCs. Our findings along with other similar findings showed that TiO 2 NPs probably have negative effects on the cell growth, prompt the cells for entry into proliferation stop, DNA damage, and trigger the aging process. Graphical abstract., (© 2022. The Author(s).)

Introduction: There are many factors and conditions that lead to cellular senescence. Replicative senescence and Hayflick phenomenon are the most important causes of cellular senescence. Senescent cells also lead to wound healing conditions resulting from injury and toxic conditions., Material and Methods: When a cell becomes senescent, it stops replication and begins to leak inflammatory signals before growth. It also alters the extracellular matrix and behavior of neighbor cells and even motivates them. This review was conducted to determine the association between senescence and bone marrow cancer., Results: The results showed that senescent cells have a short life span due to their self-destructive nature or natural removal from the body by the immune system. These signals are effective to a certain extent in regenerating the damaged cells when present in a transient state. Cellular senescence can decrease the risk of all cancers, including bone marrow cancer, ensuring that cells with significant DNA injury are prevented from replication., Conclusion: However, senescent cells increase in number as they age, which is very harmful over time. These cells extend into an older tissue for longer periods of time and form longer clusters in older tissues. Therefore, cellular senescence significantly contributes to aging., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)

Zinc oxide (ZnO) nanoparticles (NPs) are generally utilized in cosmetic goods, sheds, biosensors, and delivery of drug. As in vitro ideal systems, mesenchymal stem cells (MSCs) are used to test acute toxicity. In the present study, size-dependent cytotoxicity effects of ZnO NPs on MSCs were assessed. Bone marrow and adipose MSCs were treated with ZnO NPs with average sizes of 10-30 and 35-45 nm. The 5 and 10 µg/ml concentrations of ZnO NP were found to be the safe concentrations for the NP sizes of 10-30 and 35-45 nm, respectively. Cell-cycle analysis indicated that the small size of ZnO NPs has more negative effects on the process of cell entry to DNA synthesis when compared to the larger size. The results of the β-galactosidase test showed the promotion of the aging process in the cells treated with the smaller size of ZnO NPs. Both sizes of the NP were found to upregulate the aging-related genes NF-kB and p53 and downregulate the anti-aging gene Nanog. To sum up, the smaller size of ZnO NPs can enhance the aging process in the cells., (© 2021. The Author(s).)

Recent studies suggest that metabolic regulation may improve differentiation of cardiomyocytes derived from induced pluripotent stem cells (iPSCs). AMP-activated protein kinase (AMPK) is a master regulator of metabolic activities. We investigated whether AMPK participates in iPSC-derived cardiomyocyte differentiation. We observed that AMPK phosphorylation at Thr172 increased at day 9 but then decreased after day 11 of differentiation to cardiomyocytes. Inhibition of AMPK with compound C significantly reduced mRNA and protein expression of cardiac troponins TNNT2 and TNNI3. Moreover, sustained AMPK activation using AICAR from days 9 to 14 of differentiation increased mRNA and protein expression of both TNNT2 and TNNI3. AICAR decreased acetylation of histone 3 at Lys9 and 56 and histone 4 at Lys16 (known target sites for nuclear-localized sirtuins [SIRT1, SIRT6]), suggesting that AMPK activation enhances sirtuin activity. Sustained AMPK activation during days 9-14 of differentiation induces sirtuin-mediated histone deacetylation and may enhance cardiomyocyte differentiation from iPSCs., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Current differentiation protocols to produce cardiomyocytes from human induced pluripotent stem cells (iPSCs) are capable of generating highly pure cardiomyocyte populations as determined by expression of cardiac troponin T. However, these cardiomyocytes remain immature, more closely resembling the fetal state, with a lower maximum contractile force, slower upstroke velocity, and immature mitochondrial function compared with adult cardiomyocytes. Immaturity of iPSC-derived cardiomyocytes may be a significant barrier to clinical translation of cardiomyocyte cell therapies for heart disease. During development, cardiomyocytes undergo a shift from a proliferative state in the fetus to a more mature but quiescent state after birth. The mechanistic target of rapamycin (mTOR)-signaling pathway plays a key role in nutrient sensing and growth. We hypothesized that transient inhibition of the mTOR-signaling pathway could lead cardiomyocytes to a quiescent state and enhance cardiomyocyte maturation., Methods: Cardiomyocytes were differentiated from 3 human iPSC lines using small molecules to modulate the Wnt pathway. Torin1 (0 to 200 nmol/L) was used to inhibit the mTOR pathway at various time points. We quantified contractile, metabolic, and electrophysiological properties of matured iPSC-derived cardiomyocytes. We utilized the small molecule inhibitor, pifithrin-α, to inhibit p53 signaling, and nutlin-3a, a small molecule inhibitor of MDM2 (mouse double minute 2 homolog) to upregulate and increase activation of p53., Results: Torin1 (200 nmol/L) increased the percentage of quiescent cells (G 0 phase) from 24% to 48% compared with vehicle control ( P <0.05). Torin1 significantly increased expression of selected sarcomere proteins (including TNNI3 [troponin I, cardiac muscle]) and ion channels (including Kir2.1) in a dose-dependent manner when Torin1 was initiated after onset of cardiomyocyte beating. Torin1-treated cells had an increased relative maximum force of contraction, increased maximum oxygen consumption rate, decreased peak rise time, and increased downstroke velocity. Torin1 treatment increased protein expression of p53, and these effects were inhibited by pifithrin-α. In contrast, nutlin-3a independently upregulated p53, led to an increase in TNNI3 expression and worked synergistically with Torin1 to further increase expression of both p53 and TNNI3., Conclusions: Transient treatment of human iPSC-derived cardiomyocytes with Torin1 shifts cells to a quiescent state and enhances cardiomyocyte maturity.

Caloric restriction has been associated with increased life span and reduced aging-related disorders and reduces fibrosis in several diseases. Fibrosis is characterized by deposition of excess fibrous material in tissues and organs and is caused by aging, chronic stress, injury, or disease. Myofibroblasts are fibroblast-like cells that secrete high levels of extracellular matrix proteins, resulting in fibrosis. Histological studies have identified many-fold increases of myofibroblasts in aged organs where myofibroblasts are constantly generated from resident tissue fibroblasts and other cell types. However, it remains unclear how aging increases the generation of myofibroblasts. Here, using mouse models and biochemical assays, we show that sirtuin 6 (SIRT6) deficiency plays a major role in aging-associated transformation of fibroblasts to myofibroblasts, resulting in tissue fibrosis. Our findings suggest that SIRT6-deficient fibroblasts transform spontaneously to myofibroblasts through hyperactivation of transforming growth factor β (TGF-β) signaling in a cell-autonomous manner. Importantly, we noted that SIRT6 haploinsufficiency is sufficient for enhancing myofibroblast generation, leading to multiorgan fibrosis and cardiac dysfunction in mice during aging. Mechanistically, SIRT6 bound to and repressed the expression of key TGF-β signaling genes by deacetylating SMAD family member 3 (SMAD3) and Lys-9 and Lys-56 in histone 3. SIRT6 binding to the promoters of genes in the TGF-β signaling pathway decreased significantly with age and was accompanied by increased binding of SMAD3 to these promoters. Our findings reveal that SIRT6 may be a potential candidate for modulating TGF-β signaling to reduce multiorgan fibrosis during aging and fibrosis-associated diseases., Competing Interests: The authors declare that they have no conflicts of interest with the contents of this article., (© 2020 Maity et al.)

Aims: Isolation and identification of genes encoding putative phosphatases from Pseudomonas putida strain P13 DSM 23335., Methods and Results: By functional screening of a P. putida P13 genomic library, a number of Pho + clones were identified. Two genes were identified that encoded proteins exhibiting both phytase and sugar phosphatase activities. The proteins were 249 and 462 amino acids, with molecular masses of 26 and 50 kDa respectively. Sequence alignments revealed no significant similarities to representatives of known phosphatase or phytase gene families. However, the genes were found to have a high similarity to members of the major facilitator superfamily (MFS). Both genes were overexpressed in Escherichia coli and the corresponding partially purified recombinant enzymes were found to have significant phytate-dephosphorylating activity. The protein designated P. putida phytase 1 (Ppp1) displayed the highest activity among potential substrates studied on Na phytate, whereas Ppp2 more likely represents a sugar phosphatase than a phytase. The optimal conditions for phytate dephosphorylation were determined as 60°C and pH 4·5 (Ppp1) or pH 5·0 (Ppp2)., Conclusions: Two novel bacterial phosphatase-encoding genes, named ppp1 and ppp2, were isolated from P. putida P13 DSM 23335 by a functional screening procedure., Significance and Impact of the Study: Phosphatase-encoding genes are of great importance for industrial applications, particularly in agriculture. The identified phosphatase genes represent a new class of acid phosphatases., (© 2019 The Society for Applied Microbiology.)

Global protein synthesis is emerging as an important player in the context of aging and age-related diseases. However, the intricate molecular networks that regulate protein synthesis are poorly understood. Here, we report that SIRT6, a nuclear-localized histone deacetylase represses global protein synthesis by transcriptionally regulating mTOR signalling via the transcription factor Sp1, independent of its deacetylase activity. Our results suggest that SIRT6 deficiency increases protein synthesis in mice. Further, multiple lines of in vitro evidence suggest that SIRT6 negatively regulates protein synthesis in a cell-autonomous fashion and independent of its catalytic activity. Mechanistically, SIRT6 binds to the zinc finger DNA binding domain of Sp1 and represses its activity. SIRT6 deficiency increased the occupancy of Sp1 at key mTOR signalling gene promoters resulting in enhanced expression of these genes and activation of the mTOR signalling pathway. Interestingly, inhibition of either mTOR or Sp1 abrogated the increased protein synthesis observed under SIRT6 deficient conditions. Moreover, pharmacological inhibition of mTOR restored cardiac function in muscle-specific SIRT6 knockout mice, which spontaneously develop cardiac hypertrophy. Overall, these findings have unravelled a new layer of regulation of global protein synthesis by SIRT6, which can be potentially targeted to combat aging-associated diseases like cardiac hypertrophy., (© The Author(s) 2019. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Tumor necrosis factor-α (TNFα), one of the major proinflammatory cytokines, plays a key role in an effective immune response. However, the chronic presence of TNFα can lead to several inflammatory disorders, such as rheumatoid arthritis, psoriasis, Crohn's disease, etc. Inhibition of TNFα by pharmacological inhibitors or antibodies has proven to be effective in palliative treatment to some extent. The aim of this study was to develop an anti-TNFα antibody, which may be used as a therapeutic option to inhibit TNFα-mediated cytotoxicity. We characterized several hybridoma clones secreting monoclonal antibodies (mAbs) to human-TNFα. Four mAbs rescued L929 fibroblast cells from TNFα-triggered cell death and one of these, namely C8, was found to have the highest affinity. To gain insights into the mechanism by which mAb C8 inhibits human TNFα-mediated toxicity, the epitope corresponding to the mAb was delineated. The antigenic determinant was found to comprise of the stretch of amino acids 99-120, of which, 102-104 (glutamine, arginine, glutamic acid) form the core epitope. The observation was supported by bioinformatics analyses of an antigen/antibody complex model. In addition, the binding affinity of mAb C8 to TNFα was found to be comparable with that of infliximab, which is a commercially available anti-TNFα mAb.

Salmonella being a successful pathogen, employs a plethora of immune evasion mechanisms. This contributes to pathogenesis, persistence and also limits the efficacy of available treatment. All these contributing factors call upon for new drug targets against Salmonella. For the first time, we have demonstrated that Salmonella upregulates sirtuin 2 (SIRT2), an NAD+ dependent deacetylase in dendritic cells (DC). SIRT2 upregulation results in translocation of NFκB p65 to the nucleus. This further upregulates NOS2 transcription and nitric oxide (NO) production. NO subsequently shows antibacterial activity and suppresses T cell proliferation. NOS2 mediated effect of SIRT2 is further validated by the absence of effect of SIRT2 inhibition in NOS2-/- mice. Inhibition of SIRT2 increases intracellular survival of the pathogen and enhances antigen presentation in vitro. However, in vivo SIRT2 inhibition shows lower bacterial organ burden and reduced tissue damage. SIRT2 knockout mice also demonstrate reduced bacterial organ burden compared to wild-type mice. Collectively, our results prove the role of SIRT2 in Salmonella pathogenesis and the mechanism of action. This can aid in designing of host-targeted therapeutics directed towards inhibition of SIRT2., Competing Interests: The authors have declared that no competing interests exist.

c-Jun NH 2 -terminal kinases (JNKs) are responsive to stress stimuli and their activation regulate key cellular functions, including cell survival, growth, differentiation and aging. Previous studies demonstrate that activation of JNK requires dual phosphorylation by the mitogen-activated protein kinase kinases. However, other post-translational mechanisms involved in regulating the activity of JNK have been poorly understood. In this work, we studied the functional significance of reversible lysine acetylation in regulating the kinase activity of JNK. We found that the acetyl transferase p300 binds to, acetylates and inhibits kinase activity of JNK. Using tandem mass spectrometry, molecular modelling and molecular dynamics simulations, we found that acetylation of JNK at Lys153 would hinder the stable interactions of the negatively charged phosphates and prevent the adenosine binding to JNK. Our screening for the deacetylases found SIRT2 as a deacetylase for JNK. Mechanistically, SIRT2-dependent deacetylation enhances ATP binding and enzymatic activity of JNK towards c-Jun. Furthermore, SIRT2-mediated deacetylation favours the phosphorylation of JNK by MKK4, an upstream kinase. Our results indicate that deacetylation of JNK by SIRT2 promotes oxidative stress-induced cell death. Conversely, SIRT2 inhibition attenuates H 2 O 2 -mediated cell death in HeLa cells. SIRT2-deficient (SIRT2-KO) mice exhibit increased acetylation of JNK, which is associated with markedly reduced catalytic activity of JNK in the liver. Interestingly, SIRT2-KO mice were resistant to acetaminophen-induced liver toxicity. SIRT2-KO mice show lower cell death, minimal degenerative changes, improved liver function and survival following acetaminophen treatment. Overall, our work identifies SIRT2-mediated deacetylation of JNK as a critical regulator of cell survival during oxidative stress.

Toll-like receptors (TLRs) are a family of pattern-recognition receptors involved in innate immunity. Previous studies have shown that TLR2 inhibition protects the heart from acute stress, including myocardial infarction and doxorubicin-induced cardiotoxicity in animal models. However, the role of TLR2 in the development of aging-associated heart failure is not known. In this work, we studied aging-associated changes in structure and function of TLR2-deficient mice hearts. Whereas young TLR2-KO mice did not develop marked cardiac dysfunction, 8- and 12-month-old TLR2-KO mice exhibited spontaneous adverse cardiac remodeling and cardiac dysfunction in an age-dependent manner. The hearts of the 8-month-old TLR2-KO mice had increased fibrosis, cell death, and reactivation of fetal genes. Moreover, TLR2-KO hearts displayed reduced infiltration by macrophages, increased numbers of myofibroblasts and atrophic cardiomyocytes, and higher levels of the atrophy-related ubiquitin ligases MuRF-1 and atrogin-1. Mechanistically, TLR2 deficiency impaired the PI3K/Akt signaling pathway, leading to hyperactivation of the transcription factor Forkhead box protein O1 (FoxO1) and, in turn, to elevated expression of FoxO target genes involved in the regulation of muscle wasting and cell death. AS1842856-mediated chemical inhibition of FoxO1 reduced the expression of the atrophy-related ubiquitin ligases and significantly reversed the adverse cardiac remodeling while improving the contractile functions in the TLR2-KO mice. Interestingly, TLR2 levels decreased in hearts of older mice, and the activation of TLR1/2 signaling improved cardiac functions in these mice. These findings suggest that TLR2 signaling is essential for protecting the heart against aging-associated adverse remodeling and contractile dysfunction in mice., (© 2018 Spurthi et al.)

In the long term, diabetes profoundly affects multiple organs, such as the kidney, heart, brain, liver, and eyes. The gradual loss of function in these vital organs contributes to mortality. Nonetheless, the effects of diabetes on the lung tissue are not well understood. Clinical and experimental data from our studies revealed that diabetes induces inflammatory and fibrotic changes in the lung. These changes were mediated by TGF-β-activated epithelial-to-mesenchymal transition (EMT) signaling pathways. Our studies also found that glucose restriction promoted mesenchymal-to-epithelial transition (MET) and substantially reversed inflammatory and fibrotic changes, suggesting that diabetes-induced EMT was mediated in part by the effects of hyperglycemia. Additionally, the persistent exposure of diabetic cells to high glucose concentrations (25 mM) promoted the upregulation of caveolin-1, N-cadherin, SIRT3, SIRT7 and lactate levels, suggesting that long-term diabetes may promote cell proliferation. Taken together, our results demonstrate for the first time that diabetes induces fibrotic changes in the lung via TGF-β1-activated EMT pathways and that elevated SMAD7 partially protects the lung during the initial stages of diabetes. These findings have implications for the management of patients with diabetes.

Bacillus subtilis is a Gram positive, aerobic and motile bacterium. Biofilm formation is an important feature of this bacterium which confers resistance to antimicrobial agents. The use of new antimicrobial reagents which eliminate biofilms are important and necessary. In this study, the effect of secondary metabolites (bacteriocin) from Lactobacillus acidophilus ATCC 4356 on Bacillus subtilis BM19 in the presence and absence of HBsu which is involved in the growth of planktonic cells and biofilm formation, is reported. HBsu nucleoprotein plays several roles in different processes of Bacillus subtilis cells such as replication, transcription, cell division, recombination and repair. In this study, for the first time, the effect of HBsu on biofilm formation is presented., Results: In the absence of HBsu, purified bacteriocin from L. acidophilus ATCC 4356 was more effective in inhibiting growth of B. subtilis BM19 planktonic cells as well as biofilm formation. The presence of HBsu on the other hand led to increased biofilm formation., (Copyright © 2018. Published by Elsevier B.V.)

Sirtuins are a family of enzymes, which govern a number of cellular processes essential for maintaining physiological balance. SIRT6, a nuclear sirtuin, is implicated in the development of metabolic disorders. The role of SIRT6 in regulation of cardiac metabolism is unexplored. Although glucose is not the primary energy source of heart, defects in glucose oxidation have been linked to heart failure. SIRT6 +/- mice hearts exhibit increased inhibitory phosphorylation of PDH subunit E1α. SIRT6 deficiency enhances FoxO1 nuclear localization that results in increased expression of PDK4. We show that SIRT6 transcriptionally regulates the expression of PDK4 by binding to its promoter. SIRT6 +/- hearts show accumulation of lactate, indicating compromised mitochondrial oxidation. SIRT6 deficiency results in decreased oxygen consumption rate and concomitantly lesser ATP production. Mechanistically, SIRT6 deficiency leads to increased FoxO1-mediated transcription of PDK4. Our findings establish a novel link between SIRT6 and cardiac metabolism, suggesting a protective role of SIRT6 in maintaining cardiac homeostasis., (© 2018 Wiley Periodicals, Inc.)

Heart failure is an aging-associated disease that is the leading cause of death worldwide. Sirtuin family members have been largely studied in the context of aging and aging-associated diseases. Sirtuin 2 (SIRT2) is a cytoplasmic protein in the family of sirtuins that are NAD + -dependent class III histone deacetylases. In this work, we studied the role of SIRT2 in regulating nuclear factor of activated T-cells (NFAT) transcription factor and the development of cardiac hypertrophy. Confocal microscopy analysis indicated that SIRT2 is localized in the cytoplasm of cardiomyocytes and SIRT2 levels are reduced during pathological hypertrophy of the heart. SIRT2-deficient mice develop spontaneous pathological cardiac hypertrophy, remodeling, fibrosis, and dysfunction in an age-dependent manner. Moreover, young SIRT2-deficient mice develop exacerbated agonist-induced hypertrophy. In contrast, SIRT2 overexpression attenuated agonist-induced cardiac hypertrophy in cardiomyocytes in a cell-autonomous manner. Mechanistically, SIRT2 binds to and deacetylates NFATc2 transcription factor. SIRT2 deficiency stabilizes NFATc2 and enhances nuclear localization of NFATc2, resulting in increased transcription activity. Our results suggest that inhibition of NFAT rescues the cardiac dysfunction in SIRT2-deficient mice. Thus, our study establishes SIRT2 as a novel endogenous negative regulator of NFAT transcription factor., (© 2018 by The American Society for Biochemistry and Molecular Biology, Inc.)

Cardiomyopathy is one of the characteristic features of cancer. In this study, we establish a suitable model to study breast cancer-induced cardiomyopathy in mice. We used Ehrlich Ascites Carcinoma cells to induce subcutaneous tumor in 129/SvJ mice and studied its effect on heart function. In Ehrlich Ascites Carcinoma bearing mice, we found significant reduction in left ventricle wall thickness, ejection fraction, and fractional shortening increase in left ventricle internal diameter. We found higher muscle atrophy, degeneration, fibrosis, expression of cell-adhesion molecules and cell death in tumor-bearing mice hearts. As observed in cancer patients, we found that mTOR, a key signalling molecule responsible for maintaining cell growth and autophagy was suppressed in this model. Tumor bearing mice hearts show increased expression and nuclear localization of TFEB and FoxO3a transcription factors, which are involved in the upregulation of muscle atrophy genes, lysosomal biogenesis genes and autophagy genes. We propose that Ehrlich Ascites Carcinoma induced tumor can be used as a model to identify potential therapeutic targets for the treatment of heart failure in patients suffering from cancer-induced cardiomyopathy. This model can also be used to test the adverse consequences of cancer chemotherapy in heart.

Glycogen synthase kinase 3 (GSK3) is a critical regulator of diverse cellular functions involved in the maintenance of structure and function. Enzymatic activity of GSK3 is inhibited by N-terminal serine phosphorylation. However, alternate post-translational mechanism(s) responsible for GSK3 inactivation are not characterized. Here, we report that GSK3α and GSK3β are acetylated at Lys246 and Lys183, respectively. Molecular modeling and/or molecular dynamics simulations indicate that acetylation of GSK3 isoforms would hinder both the adenosine binding and prevent stable interactions of the negatively charged phosphates. We found that SIRT2 deacetylates GSK3β, and thus enhances its binding to ATP. Interestingly, the reduced activity of GSK3β is associated with lysine acetylation, but not with phosphorylation at Ser9 in hearts of SIRT2-deficient mice. Moreover, GSK3 is required for the anti-hypertrophic function of SIRT2 in cardiomyocytes. Overall, our study identified lysine acetylation as a novel post-translational modification regulating GSK3 activity., Competing Interests: MS, SM, SM, CK, DW, MG, MS, NS No competing interests declared, (© 2018, Sarikhani et al.)

Sepsis, a leading cause of death in intensive care units, is primarily caused due to an exaggerated immune response. The hyperactive inflammatory response mediated by immune cells against infectious organisms and their toxins results in host cell death and tissue damage, the hallmarks of septic shock. Therefore, molecules that modulate inflammatory responses are attractive therapeutic targets for sepsis. Nitric oxide (NO) is a signaling molecule, which is implicated in regulating diverse immune functions. Although, the protective roles of NO in infectious diseases are well documented, its importance in sepsis is controversial. In the present study, the effects of intra-peritoneal injection of mice with Salmonella Typhimurium, a Gram-negative intracellular pathogen, were studied which leads to a rapid upregulation of serum cytokines and infiltration of neutrophils to the peritoneal cavity. Surprisingly, the induction of inflammatory cytokines and chemokines, e.g. IL6 and CCL2, and the infiltration of neutrophils into the peritoneal cavity are mitigated in mice lacking Nitric oxide synthase 2 (NOS2). The reduced inflammatory response in Nos2 -/- mice is accompanied by greater bacterial burden in the peritoneal cavity, lower thymic atrophy, higher liver damage and cardiovascular dysfunction followed by decreased survival. However, no significant differences are observed in other responses between C57BL/6 wild type (WT) and Nos2 -/- mice: induction of glucocorticoids, phagocytic ability and apoptosis of peritoneal cells. This study clearly highlights the NOS2-dependent and -independent responses in this mouse model of peritonitis induced sepsis. Importantly, pre-treatment of Nos2 -/- mice with DETA-NO, a NO donor, upon infection, restores neutrophil recruitment, reduces bacterial numbers in the peritoneal cavity, improves liver and cardio-vascular function and enhances survival. Interestingly, DETA-NO treatment does not significantly increase the survival of infected WT mice. The implications of these results and the complex roles of NO as a target molecule during sepsis are discussed., (Copyright © 2018 Elsevier Inc. All rights reserved.)