Current paradigms and future challenges in harnessing gut bacterial symbionts of insects for biodegradation of plastic wastes.

The ubiquitous incorporation of plastics into daily life, coupled with inefficient recycling practices, has resulted in the accumulation of millions of metric tons of plastic waste, that poses a serious threat to the Earth's sustainability. Plastic pollution, a global problem, disrupts the ecological balance and endangers various life forms. Efforts to combat plastic pollution are underway, with a promising avenue being biological degradation facilitated by certain insects and their symbiotic gut microorganisms, particularly bacteria. This review consolidates existing knowledge on plastic degradation by insects and their influence on gut microbiota. Additionally, it delves into the potential mechanisms employed by insects in symbiosis with gut bacteria, exploring the bioconversion of waste plastics into value-added biodegradable polymers through mineralization. These insights hold significant promise for the bio-upcycling of plastic waste, opening new horizons for future biomanufacturing of high-value chemicals from plastic-derived compounds. Finally, we weigh the pros and cons of future research endeavors related to the bioprospection of plastic-degrading bacteria from underexplored insect species. We also underscore the importance of bioengineering depolymerases with novel characteristics, aiming for their application in the remediation and valorization of waste plastics.

Analysis of Dysferlin Direct Interactions with Putative Repair Proteins Links Apoptotic Signaling to Ca2+ Elevation via PDCD6 and FKBP8.

Quantitative surface plasmon resonance (SPR) was utilized to determine binding strength and calcium dependence of direct interactions between dysferlin and proteins likely to mediate skeletal muscle repair, interrupted in limb girdle muscular dystrophy type 2B/R2. Dysferlin canonical C2A (cC2A) and C2F/G domains directly interacted with annexin A1, calpain-3, caveolin-3, affixin, AHNAK1, syntaxin-4, and mitsugumin-53, with cC2A the primary target and C2F lesser involved, overall demonstrating positive calcium dependence. Dysferlin C2 pairings alone showed negative calcium dependence in almost all cases. Like otoferlin, dysferlin directly interacted via its carboxy terminus with FKBP8, an anti-apoptotic outer mitochondrial membrane protein, and via its C2DE domain with apoptosis-linked gene (ALG-2/PDCD6), linking anti-apoptosis with apoptosis. Confocal Z-stack immunofluorescence confirmed co-compartmentalization of PDCD6 and FKBP8 at the sarcolemmal membrane. Our evidence supports the hypothesis that prior to injury, dysferlin C2 domains self-interact and give rise to a folded, compact structure as indicated for otoferlin. With elevation of intracellular Ca2+ in injury, dysferlin would unfold and expose the cC2A domain for interaction with annexin A1, calpain-3, mitsugumin 53, affixin, and caveolin-3, and dysferlin would realign from its interactions with PDCD6 at basal calcium levels to interact strongly with FKBP8, an intramolecular rearrangement facilitating membrane repair.

In situ engineering of Au-Ag alloy embedded PEDOT nanohybrids at a solvent/non-solvent interface for the electrochemical enzyme-free detection of histamine.

Steadfast efforts have been made to develop novel materials and incorporate them into functional devices for practical applications, pushing the research on electroactive materials to the forefront of nano electronics. Liquid/liquid interface-assisted polymerization offers a scalable methodology to fabricate hybrid materials with multifunctional applications, in contrast to the conventional and ubiquitous routes. Here, we explored this efficient and versatile approach toward the in situ tailoring of Au-Ag alloy nanostructures with a conducting polymer, poly(3,4-ethylene-dioxythiophene) (PEDOT). With the appropriate choice of organic and inorganic phases for the distribution of monomer and oxidant, the miscibility restraints of the reactants in a single phase were alleviated. Effective nanostructure tuning of highly crystalline and electroactive PEDOT/Au-Ag alloy has been achieved by varying the molar ratio of Au3+/Ag+ in the reaction mixture. The as-synthesized composite is further explored to detect neuromodulator histamine (HA), which displays high sensitivity with a limit of detection (LOD) of 1.5 nM, and selectivity even in the presence of various interfering analogs of 10-fold concentration. Subsequently, density functional theory (DFT) simulations are employed to assess the mode of interaction between HA and the electroactive surfaces. The competency to detect HA in preserved food entails its potential in food spoilage monitoring. Furthermore, the detection of histamine generated by sub-cultured human neuronal cells SH-SY5Y proves its practical viability in health monitoring devices.

Ligand-protected nanoclusters and their role in agriculture, sensing and allied applications.

Nano biotechnology, when coupled with green chemistry, can revolutionize human life because of the vast opportunities and benefits it can offer to the quality of human life. Luminescent metal nanoclusters (NCs) have recently developed as a potential research area with applications in different areas like medical, imaging, sensing etc. Recently these new candidates have proved to be beneficial in the food supply chain enabling controlled release of nutrients, pesticides and as nanosensors for the detection of contaminants and play roles in healthy food storage and maintaining food quality. An assortment of nanomaterials has been employed for these applications and reviews have been published on the use of nanotechnology in agriculture. Ligand-protected metal nanoclusters are a distinctive class of small organic-inorganic nanostructures that garnered immense research interest in recent years owing to their stability at specific "magic size" compositions along with tunable properties that make them promising candidates for a wide range of nanotechnology-based applications. This review tries to consolidate the recent developments in the area of ligand-protected nanoclusters in connection with the detection of pesticides, food contaminants, heavy metal ions and plant growth monitoring for healthy agricultural practices. Its antimicrobial activity to manage the microbial contamination is highlighted. The review also throws light on the various perspectives by which food production and allied areas will be transformed in future.

Valorization Potential of a Novel Bacterial Strain, Bacillus altitudinis RSP75, towards Lignocellulose Bioconversion: An Assessment of Symbiotic Bacteria from the Stored Grain Pest, Tribolium castaneum.

Bioconversion of lignocellulose into renewable energy and commodity products faces a major obstacle of inefficient saccharification due to its recalcitrant structure. In nature, lignocellulose is efficiently degraded by some insects, including termites and beetles, potentially due to the contribution from symbiotic gut bacteria. To this end, the presented investigation reports the isolation and characterization of cellulolytic bacteria from the gut system of red flour beetle, Tribolium castaneum. Out of the 15 isolated bacteria, strain RSP75 showed the highest cellulolytic activities by forming a clearance zone of 28 mm in diameter with a hydrolytic capacity of ~4.7. The MALDI-TOF biotyping and 16S rRNA gene sequencing revealed that the strain RSP75 belongs to Bacillus altitudinis. Among the tested enzymes, B. altitudinis RSP75 showed maximum activity of 63.2 IU/mL extract for xylanase followed by ß-glucosidase (47.1 ± 3 IU/mL extract) which were manifold higher than previously reported activities. The highest substrate degradation was achieved with wheat husk and corn cob powder which accounted for 69.2% and 54.5%, respectively. The scanning electron microscopy showed adhesion of the bacterial cells with the substrate which was further substantiated by FTIR analysis that depicted the absence of the characteristic cellulose bands at wave numbers 1247, 1375, and 1735 cm-1 due to hydrolysis by the bacterium. Furthermore, B. altitudinis RSP75 showed co-culturing competence with Saccharomyces cerevisiae for bioethanol production from lignocellulose as revealed by GC-MS analysis. The overall observations signify the gut of T. castaneum as a unique and impressive reservoir to prospect for lignocellulose-degrading bacteria that can have many biotechnological applications, including biofuels and biorefinery.

Recent advances in the bioprospection and applications of chitinolytic bacteria for valorization of waste chitin.

One of the most abundant natural polymers on earth, chitin is a fibrous and structural polysaccharide, composed of N-acetyl-D-glucosamine. The biopolymer is the major structural constituent of fungi, arthropods, mollusks, nematodes, and some algae. The biodegradation of chitin is largely manifested by chitinolytic enzyme secreting organisms including bacteria, insects, and plants. Among them, bacterial chitinases represent the most promising, inexpensive, and sustainable source of proteins that can be employed for industrial-scale applications. To this end, the presented review comes at a timely moment to highlight the major sources of chitinolytic bacteria. It also discusses the potential pros and cons of prospecting bacterial chitinases that can be easily manipulated through genetic engineering. Additionally, we have elaborated the recent applications of the chitin thereby branding chitinases as potential candidates for biorefinery and biomedical research for eco-friendly and sustainable management of chitin waste in the environment.

Applications of in silico methods to analyze the toxicity and estrogen receptor-mediated properties of plant-derived phytochemicals.

A myriad of phytochemicals may have potential to lead toxicity and endocrine disruption effects by interfering with nuclear hormone receptors. In this examination, the toxicity and estrogen receptor-binding abilities of a set of 2826 phytochemicals were evaluated. The endpoints mutagenicity, carcinogenicity (both CAESAR and ISS models), developmental toxicity, skin sensitization and estrogen receptor relative binding affinity (ER_RBA) were studied using the VEGA QSAR modeling package. Alongside the predictions, models were providing possible information for applicability domains and most similar compounds as similarity sets from their training sets. This information was subjected to perform the clustering and classification of chemicals using Self-Organizing Maps. The identified clusters and their respective indicators were considered as potential hotspot structures for the specified data set analysis. Molecular screening interpretations of models were exhibited accurate predictions. Moreover, the indication sets were defined significant clusters and cluster indicators with probable prediction labels (precision). Accordingly, developed QSAR models showed good predictive abilities and robustness, which observed from applicability domains, representation spaces, clustering and classification schemes. Furthermore, the designed new path could be useful as a valuable approach to determine toxicity levels of phytochemicals and other environmental pollutants and protect the human health.

Elucidation of endocrine-disrupting polychlorinated biphenyls binding potency with steroidogenic genes: Integration of in silico methods and ensemble docking approaches.

A myriad of polychlorinated biphenyls (PCBs) may have potential to reproductive axis and endocrine disruptions. PCBs mostly breach the cholesterol biotransformation in mitochondria through interfering with steroidogenic genes and lead to adverse consequences in steroidogenesis; however, studies are scanty. In this examination, the combinations of quantitative structure-activity relationship (QSAR) modeling and ensemble docking approaches was performed to envisage structural properties of PCBs that influence the developmental toxicity, estrogen receptor-mediated impacts, and to provide a better comprehension of binding levels between PCBs, steroidogenic acute regulatory protein (StAR) and cholesterol side-chain cleavage enzyme (CYP11A1). Prognostic QSAR models were illustrated with good robustness and predictive ability. Models provide extensive data on applicability domain and similarities between PCBs and training set compounds was used to implement for clustering and classification of toxic PCBs by employing Self-Organizing Maps. Docking and interaction profiles interpretations provided various insights into the structural features of PCBs that influence the cholesterol binding to StAR and CYP11A1 domains. The non-polar, atomic π - stacking and halogen bonds of PCBs with novel hotspots residues of StAR and CYP11A1 was indicated subtle conformational changes that barrier to cholesterol binding and/or locks to cholesterol ejection from Ω1-loop of StAR, and inhibits cholesterol to pregnenolone biosynthesis by CYP11A1; thus, these are probably revealed as block-cluster mechanisms. These outcomes are potential to be useful to predict developmental toxicity, endocrine disruption potencies and anti-steroidogenic activities of other environmental pollutants and provided sorted pinpoints for further evaluation of interaction mechanisms of PCBs with other sterodogenic genes.

Effectiveness of a Structured Teaching Program on Knowledge and Attitudes Toward HIV among Young Women in India.

India has the third highest number of persons living with HIV in the world. We evaluated the effectiveness of a structured teaching program on HIV-related knowledge and attitudes among 600 young women, aged 20-40 years, in India. This study employed a quasi-experimental pre-post design. Significant differences were found in the level of the participants' knowledge of (t = 78.89, p < .05) and attitudes toward (t = 64.14, p < .05) HIV after the intervention. Our findings suggest that the structured teaching program could be utilized in increasing young women's knowledge of, and improving their attitudes about, HIV.

Molecular activities and ligand-binding specificities of StAR-related lipid transfer domains: exploring integrated in silico methods and ensemble-docking approaches.

In this study, cholesterol biotransformation gene-set of human steroidogenic acute regulatory protein-related lipid transfer (START) domains were evaluated from high-throughput gene screening approaches. It was shown that STARD1, STARD3 and STARD4 proteins are better effective transporters of cholesterol than STARD5 and STARD6 domains. Docking studies show a strong agreement with gene ontology enrichment data. According to both complementary strategies, it was found that only STARD1, STARD3 and STARD4 are potentially involved in cholesterol biotransformation in mitochondria through Ω1-loop of C-terminal α4-helical domain. Ensemble docking assessment for a set of selected chemicals of protein-chemical networks has shown possible binding probabilities with START domains. Among those, reproductive toxicity evoked drugs (mifepristone), insecticides (rotenone), tobacco pulmonary carcinogens (benzo(a)pyrene) and endocrine disruptor chemicals (EDCs) including perfluorooctanesulfonic acid (PFOS) and aflatoxin B1 (AFB1) potentially bound with novel hotspot residues of the α4-helical domain. Compound representation space and clustering approaches reveal that the START proteins show more sensitivity with these lead scaffolds, so they could provide probable barrier assets in cholesterol and steroidogenic acute regulatory (StAR) binding and leads adverse consequences in steroidogenesis. These findings indicate potential START domains and their binding levels with toxic chemicals; sorted viewpoints could be useful as a promising way to identify chemicals with related steroidogenisis impacts on human health.

ERK1/2 activated PHLPP1 induces skeletal muscle ER stress through the inhibition of a novel substrate AMPK.

Nutritional abundance associated with chronic inflammation and dyslipidemia impairs the functioning of endoplasmic reticulum (ER) thereby hampering cellular responses to insulin. PHLPP1 was identified as a phosphatase which inactivates Akt, the master regulator of insulin mediated glucose homeostasis. Given the suggestive role of PHLPP1 phosphatase in terminating insulin signalling pathways, deeper insights into its functional role in inducing insulin resistance are warranted. Here, we show that PHLPP1 expression is enhanced in skeletal muscle of insulin resistant rodents which also displayed ER stress, an important mediator of insulin resistance. Using cultured cells and PHLPP1 knockdown mice, we demonstrate that PHLPP1 facilitates the development of ER stress. Importantly, shRNA mediated ablation of PHLPP1 significantly improved glucose clearance from systemic circulation with enhanced expression of glucose transporter 4 (GLUT-4) in skeletal muscle. Mechanistically, we show that endogenous PHLPP1 but not PP2Cα interacts with and directly dephosphorylates AMPK Thr172 in myoblasts without influencing its upstream kinase, LKB1. While the association between endogenous PHLPP1 and AMPK was enhanced in ER stressed cultured cells and soleus muscle of high fat diet fed mice, the basal interaction between PP2Ac and AMPK was minimally altered. Further, we show that PHLPP1α is phosphorylated by ERK1/2 at Ser932 under ER stress which is required for its ability to interact with and dephosphorylate AMPK and thereby induce ER stress. Taken together, our data position PHLPP1 as a key regulator of ER stress.

Integration of in silico approaches to determination of endocrine-disrupting perfluorinated chemicals binding potency with steroidogenic acute regulatory protein.

A myriad of perfluorinated compounds (PFCs) have the ability to interfere with steroidogenic acute regulatory (StAR) protein. Consequently, PFCs breaches cholesterol biotransformation in mitochondria and cause fatal consequences in steroidogenesis, however, these were poorly characterized. In the present study, we have evaluated toxic potencies, nuclear mediated probabilities and interaction profiles with StAR of PFCs using computational system biology tools. Toxicity endpoints revealed that PFCs contain high carcinogenicity, developmental toxicity, skin sensitization effects with low mutagenic activity. Consensus qualitative nuclear receptor agonist models show higher probability rates towards ER and PPAR-γ receptor than AR and AhR models were observed. To poise the subtle fluctuations of actual predictions, balanced accuracy and MCC were computed, and they signify perfect correlation ranges in all models. Screening studies resulting protein-ligand interaction profiles showed that residues Asn148, Asn150, Glu169, Ala171, Arg182, Phe184, Arg188, Trp241, Thr263 and Phe267 were identified as novel hotspots, participated in halogen bonds, H-bonds, atomic π-stacking, π-cation interactions and salt-bridges formation. Thus, the additional bonds contribute conformer stability that holds the protein structure at flexible state, so that PFCs acts as a barrier to cholesterol binding. From docking outcomes, representation space was created, that specifies high and medium StAR binders were occupied in toxic endpoints space with active concern. PFCs restrain molecular features and mitochondrial membrane disruption functions were revealed by efficient toxicogenomics studies. These data indicate toxicity and StAR protein binding levels of PFCs, sorted pinpoints could be useful in a promising way to know the other environmental pollutants and health risks.

In-house made nucleofection buffer for efficient and cost effective transfection of RAW 264.7 macrophages.

Electroporation is the most widely employed method of gene transfer into macrophages which are hard to transfect. RAW 264.7 is a widely used cell line for studying macrophage responses. Electroporation of RAW 264.7 cells with commercial reagents although very efficient is expensive necessitating the development of cost effective alternatives. In this study, we have formulated an economical electroporation buffer for electroporation of RAW 264.7 cells compatible with commercial nucleofector apparatus. We observed that supplementation of membrane fusogenic agents such as Ficoll, PEG and membrane resealing agent, poloxamer P188, enhanced the transfection efficiency of macrophages to a level comparable to the commercially available solutions thereby providing us a cost effective solution for genetic manipulation of macrophages especially in large numbers.

LPS depletes PHLPP levels in macrophages through the inhibition of SP1 dependent transcriptional regulation.

We have previously reported that bacterial endotoxin LPS attenuates expression of PHLPP, a ser/thr phosphatase, at both transcript and protein levels in different immune cells, however the underlying molecular mechanism is unknown and is of significant interest. Here, in line with the decreased transcript levels upon LPS treatment, we observed that LPS caused significant reduction in PHLPP promoter activity. We observed that SP1, a transcription factor frequently associated with inflammation, was recruited to the PHLPP promoter region. Ectopic expression of SP1 enhanced both transcript and protein levels of PHLPP while knockdown of SP1 or pharmacological inhibition of SP1 DNA binding by mithramycin reduced PHLPP expression. Moreover, over-expression of SP1 co-activators CBP/p300 augmented SP1 driven PHLPP promoter activity. Of note, LPS treatment depleted SP1 and CBP protein levels due to which recruitment of SP1 to PHLPP promoter was reduced. Further, we found that re-introduction of SP1 restored promoter activity and transcript levels of PHLPP in LPS stimulated cells. Collectively, our data revealed the molecular mechanism underlying the regulation of PHLPP expression during LPS induced macrophage inflammatory response.

Deubiquitinase USP12 promotes LPS induced macrophage responses through inhibition of IκBα.

Post translational modifications, ubiquitination and its reversal by deubiquitination play an important role in regulating innate immune system. USP12 is a poorly studied deubiquitinase reported to regulate T-cell receptor signalling however the functional role of USP12 in macrophages, the principal architects of inflammation, is unknown. Thus, in this study we probed the involvement of USP12 in macrophage mediated inflammatory responses using bacterial endotoxin, LPS, as the model system. Here, we observed that the expression of USP12 was altered in time dependent manner in LPS stimulated RAW 264.7 macrophages at both mRNA and protein levels as revealed by qPCR and western blot analysis, respectively. Further analysis showed that LPS reduced the levels of Sp1 which enhanced the transcriptional levels of USP12. We observed that siRNA mediated ablation of USP12 expression in mouse macrophages suppressed the induction of LPS-induced iNOS and IL-6 expression but failed to alter IFN-ß synthesis, oxidative stress and phagocytic ability of macrophages. Mechanistic analysis suggest that USP12 may be required for the activation of NFκB pathway as knockdown of USP12 reduced the inhibitory phosphorylation of IκBα, a well characterized inhibitor of NFκB nuclear translocation. Further, USP12 was observed to be required for LPS elicited phosphorylation of ERK1/2 and p38. Collectively, our data suggest that USP12 may be a key mediator of LPS stimulated macrophage responses.

Sol-Gel Synthesis and Characterizations of CoMoO4 Nanoparticles: An Efficient Photocatalytic Degradation of 4-Chlorophenol.

Cobalt molybdate CoMoO4 nanoparticles (NPs) were successfully synthesized using cobalt nitrate, ammonium molybdate, citric acid and ethyl cellulose by a simple sol-gel method. Structural, morphological, optical and magnetic properties of the obtained powder were characterized by powder X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectra, high resolution scanning electron microscope (HR-SEM), energy dispersive X-ray (EDX), UV-Visible diffuse reflectance spectra (DRS), photoluminescence (PL) spectra and vibrating sample magnetometer (VSM). XRD results indicated that the resultant powder was pure single phase crystalline with monoclinic structure. FT-IR spectra indicate the type of bonds between metals and oxygen. HR-SEM images shows that the morphology of the powder consist with well defined nanoparticles (NPs) structure. VSM results showed antiferromagnetic behavior. Photo-catalytic activity of CoMoO4 nanoparticles (NPs) was performed. The addition of TiO2 catalyst enhanced the photo-catalytic activity of CoMoO4 nanoparticles (NPs). The catalysts CoMoO4, Ti02 and mixed oxide catalyst CoMoO4-TiO2 nano- composites (NCs) were tested for the photo-catalytic degradation (PCD) of 4-chlorophenol (4-CP). It was found that the PCD efficiency of CoMoO4-TiO2 NCs was higher (97.5%) than that of pure CoMoO4 (88.0%) and TiO2 (94.0%) catalysts.

Synthesis and Characterization of Fe2(MoO4)3Nano-Photocatalyst by Simple Sol-Gel Method.

Ferric molybdate (Fe2(MoO4)3) nanorods (NRs) were successfully synthesized using metal nitrates, citric acid and ethyl cellulose by a simple sol-gel method. Structural, morphological, optical and magnetic properties of the obtained powder were characterized by powder X-ray diffraction (XRD), Fourier transform infrared (FT-IR) spectra, high resolution scanning electron microscope (HR-SEM), energy dispersive X-ray (EDX), UV-Visible diffuse reflectance spectra (DRS), photoluminescence (PL) spectra and vibrating sample magnetometer (VSM). XRD results indicated that the resultant powder was pure single phase crystalline with monoclinic structure. FT-IR spectra indicate the type of bonds between metals and oxygen. HR-SEM images shows that the morphology of the powder consist with well defined nanorods (NRs) structure. VSM results showed ferromagnetic behavior. The addition of TiO2 catalyst, it enhanced the photo-catalytic activity of Fe2(MoO4)3. The mixed oxide catalyst of Fe2(MoO4)3-TiO2 nano-composites (NCs) were also tested for the photocatalytic degradation (PCD) of 4-chlorophenol (4-CP). It was found that the PCD efficiency of Fe2(MoO4)3 NCs is higher than pure Fe2(MoO4)3 and TiO2 catalysts.

Comparative Studies of Spinel MnFe2O4 Nanostructures: Structural, Morphological, Optical, Magnetic and Catalytic Properties.

Spinel MnFe2O4 nanostructures with two different morphologies such as nanoflakes (NFs) and nanoparticles (NPs) were synthesized by a simple microwave-assisted combustion (MACM) and conventional combustion (CCM) methods respectively. Powder X-ray diffraction (XRD) study showed that the samples have pure cubic spinel phase and the average crystallite size is found to be 15.13 ± 2 nm and 24.38 ± 13 nm for NFs and NPs respectively. The calculated lattice parameter values of the samples NFs and NPs are 8.476 and 8.474 Å respectively. The morphology of the samples was recorded using high resolution scanning electron microscope (HR-SEM) analysis and was found that nanoflakes and nanoparticles morphologies by MACM and CCM methods respectively. Energy dispersive X-ray (EDX) results showed that the composition of the elements was relevant as expected from these combustion methods. The optical properties of the as-prepared nanostructures were also investigated by UV-Visible Diffuse Reflectance Spectroscopy (DRS) and Photoluminescence (PL) Spectroscopy. The energy band gap (E(g)) of the sample NFs is 1.78 eV, whereas the sample NPs has 1.75 eV. The magnetic properties were investigated using the Vibrating Sample Magnetometer (VSM) at room temperature and the hysteresis loops confirmed the super-paramagnetic behavior for both samples with saturation magnetization (M(s)) values in the range of 59.29 ± 11 and 66.78 ± 06 emu/g for the samples NFs and NPs respectively. The oxidation of benzyl alcohol into benzaldehyde reached a maximum of 87.65% for MnFe2O4 NFs, whereas for MnFe2O4 NPs, the conversion was only 69.43% with 100% selectivity.

A novel immunomodulatory function of PHLPP1: inhibition of iNOS via attenuation of STAT1 ser727 phosphorylation in mouse macrophages.

PHLPP1 is a novel tumor suppressor, but its role in the regulation of innate immune responses, which are frequently dysregulated in cancer, is unexplored. Here, we report that LPS attenuated PHLPP1 expression at mRNA and protein levels in immune cells, suggesting its involvement in immune responses. To test this, we overexpressed PHLPP1 in RAW 264.7 macrophages and observed a dramatic reduction in LPS/IFN-γ-induced iNOS expression. Conversely, silencing of PHLPP1 by siRNA or by shRNA robustly augmented LPS/IFN-γ-induced iNOS expression. qPCR and iNOS promoter reporter experiments showed that PHLPP1 inhibited iNOS transcription. Mechanistic analysis revealed that PHLPP1 suppressed LPS/IFN-γ-induced phosphorylation of ser727 STAT1; however, the underlying mechanisms differed. PHLPP1 reduced IFN-γ-stimulated but not LPS-induced ERK1/2 phosphorylation, and inhibition of ERK1/2 abolished IFN-γ-induced ser727 STAT1 phosphorylation and iNOS expression. In contrast, PHLPP1 knockdown augmented LPS-induced but not IFN-γ-elicited p38 phosphorylation. Blockade of p38 abolished LPS-stimulated phosphorylation of ser727 STAT1 and iNOS expression. Furthermore, PHLPP1 suppressed LPS-induced phosphorylation of tyr701 STAT1 by dampening p38-dependent IFN-ß feedback. Collectively, our data demonstrate for the first time that PHLPP1 plays a vital role in restricting innate immune responses of macrophages, and further studies may show it to be a potential therapeutic target within the context of dysregulated macrophage activity.

Alterations in some dehydrogenase profiles of sciatectomized toad gastrocnemius muscle-metabolic modifications by malate.

Sciatectomized toad gastrocnemius has shown a progressive loss in lactate (LDH), succinate (SDH) and malate (MDH) dehydrogenase activities and elevation of glutamate dehydrogenase (GDH) activity during post-neurectemic days. The possible role of malate in the restoration of metabolic homeostasis in denervated muscle is discussed.