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Organophosphates (OPs) are found in hundreds of important products used worldwide; however, they have been associated with adverse long-term health consequences ranging from neurodevelopmental deficits to age-related neurological diseases. OP exposure has also been implicated in Gulf War Illness; a cluster of medically unexplained chronic symptoms estimated to affect 25–32% of veterans of the Persian Gulf war in 1991. The development of multiple types of chronic illnesses in these veterans at an early age compared to the general population has led to the suggestion that they are experiencing signs of premature or accelerated aging. The process of cellular senescence and the development of the senescence-associated secretory phenotype (SASP) is believed to lead to chronic inflammation, chronic illnesses, as well as accelerated biological aging, and a role of environmental exposures in these processes has been suggested, but not extensively studied to date. In the studies described here, we evaluated the persistent effects of a single (acute) exposure of a representative nerve agent OP, diisopropylfluorophosphate (DFP) 4.0 mg/kg on cognitive function, noncognitive behaviors, cellular senescence markers and proinflammatory cytokines in the mouse brain. The results indicated modest, but persistent DFP-related impairments in spatial learning and working memory, but not contextual or cued fear conditioning. DFP exposure was also not associated with negative effects on weight or impairments of the various noncognitive (e.g., motor function or exploratory activity) behavioral assessments. Both histology and quantitative PCR experiments indicated that DFP was associated with persistent alterations in several senescence markers and proinflammatory cytokines in brain regions that are relevant to the performance of the memory-related tasks (e.g., hippocampus, prefrontal cortex). The results thus suggest that single acute exposures to OPs like DFP can lead to persistent impairments in specific domains of cognition that may be related to alterations in cellular senescence and inflammaging in the brain.

The ability of Mycobacterium tuberculosis (Mtb) to tolerate nitric oxide (•NO) and superoxide (O2•−) produced by phagocytes contributes to its success as a human pathogen. Recombination of •NO and O2•− generates peroxynitrite (ONOO−), a potent oxidant produced inside activated macrophages causing lethality in diverse organisms. While the response of Mtb toward •NO and O2•− is well established, how Mtb responds to ONOO− remains unclear. Filling this knowledge gap is important to understand the persistence mechanisms of Mtb during infection. We synthesized a series of compounds that generate both •NO and O2•−, which should combine to produce ONOO−. From this library, we identified CJ067 that permeates Mtb to reliably enhance intracellular ONOO− levels. CJ067-exposed Mtb strains, including multidrug-resistant (MDR) and extensively drug-resistant (XDR) clinical isolates, exhibited dose-dependent, long-lasting oxidative stress and growth inhibition. In contrast, Mycobacterium smegmatis (Msm), a fast-growing, non-pathogenic mycobacterial species, maintained redox balance and growth in response to intracellular ONOO−. RNA-sequencing with Mtb revealed that CJ067 induces antioxidant machinery, sulphur metabolism, metal homeostasis, and a 4Fe–4S cluster repair pathway (suf operon). CJ067 impaired the activity of the 4Fe–4S cluster-containing TCA cycle enzyme, aconitase, and diminished bioenergetics of Mtb. Work with Mtb strains defective in SUF and IscS involved in Fe–S cluster biogenesis pathways showed that both systems cooperatively protect Mtb from intracellular ONOO− in vitro and inducible nitric oxide synthase (iNOS)-dependent growth inhibition during macrophage infection. Thus, Mtb is uniquely sensitive to intracellular ONOO− and targeting Fe–S cluster homeostasis is expected to promote iNOS-dependent host immunity against tuberculosis (TB).

Introduction: Depression is one of the leading causes of morbidity worldwide. It can impact various biomarkers, including proteins, monoamines, and lipids. Lipids play a critical role in regulating cellular function by influencing transport, anchoring, and providing structural support. The present study aimed to explore any correlation between lipid profiles and depression. Aim: To investigate the correlation between different lipid profile parameters (serum triglycerides, total cholesterol, serum Highdensity Lipoproteins (HDL), and Low-density Lipoproteins (LDL) levels) and the severity of depression. Materials and Methods: A hospital-based, cross-sectional study was conducted at the Department of Psychiatry, Faculty of Medicine and Health Sciences, SGT University, Gurugram, Haryana, India, from June 2020 to June 2021. A total of 200 participants aged 18-65 years, diagnosed with depression according to the ICD-10 classification, were enrolled. Clinical assessments were conducted, and the Hamilton Depression Rating Scale (HAM-D) (17 items) and Beck's Depression Inventory II (BDI II) (21-item scale) were used. Lipid parameters were obtained from fasting blood samples. The Wilcoxon’sMann-Whitney U test was applied to assess the association between gender distribution and lipid profiles. Spearman's correlation test was used to analyse the correlation between HAM-D and BDI scores with lipid profiles. Statistical significance was set at a p-value ≤0.05. Results: The mean age of the participants was 36.05±11.24 years. Correlations between HAM-D levels and lipid profiles revealed that all lipid parameters, except for HDL (which showed a negative correlation), were positively associated with HAM-D scores. These correlations were statistically significant (p ≤0.05). Similarly, a positive correlation was observed between all lipid levels and BDI scores, except for HDL, which exhibited a negative correlation. These correlations were also statistically significant (p≤0.05). Conclusion: Higher levels of LDL cholesterol were found to be associated with elevated depression rating scale scores and increased susceptibility to depression. On the other hand, HDL cholesterol demonstrated protective effects against depression. Adopting a healthy lifestyle, implementing dietary measures, and addressing stressors promptly may help prevent depressive symptoms.

Early pregnancy diagnosis is vital for economic sustainability of dairy farms and maintaining the reproductive efficiency of the herd. There are many techniques including progesterone assay, pregnancy specific proteins and interferon stimulated genes have been explored for early pregnancy diagnosis but, they are associated with varying level of efficacy. In the present experiment, interferon stimulated gene (Myxovirus resistance gene 2/MX2) expression pattern was used as a potential biomarker for early pregnancy in cattle. The association of MX2 gene expression in relation to progesterone assay was studied to explore its potential use as biomarker of early pregnancy. The plasma progesterone concentration in conceived animals on day 7 (2.26±0.19 ng/ml), 17 (5.42±0.35 ng/ml) and 21(6.38±0.39 ng/ml) was recorded to be significantly higher as compared to respective values in non-conceived animals, i.e. 1.55±0.09 ng/ml, 4.14±0.14 ng/ml and 0.81±0.06 ng/ml. The sudden decrement in plasma progesterone concentration after day 17th discriminates conceived and non-conceived animals. MX2 expression levels were observed to spike in blood due to release of interferon tau (τ) after implantation of embryo. The relative mRNA expression of MX2 gene showed a 9.5 to 28.64-fold higher expression on 17 days post insemination in pregnant animals as compared to non-pregnant animals. Thus, MX2 gene can be used as a reliable biomarker for the early detection of pregnancy.

Hierarchical Topic Modeling is the probabilistic approach for discovering latent topics distributed hierarchically among the documents. The distributed topics are represented with the respective topic terms. An unambiguous conclusion from the topic term distribution is a challenge for readers. The hierarchical topic labeling eases the challenge by facilitating an individual, appropriate label for each topic at every level. In this work, we propose a BERT-embedding inspired methodology for labeling hierarchical topics in short text corpora. The short texts have gained significant popularity on multiple platforms in diverse domains. The limited information available in the short text makes it difficult to deal with. In our work, we have used three diverse short text datasets that include both structured and unstructured instances. Such diversity ensures the broad application scope of this work. Considering the relevancy factor of the labels, the proposed methodology has been compared against both automatic and human annotators. Our proposed methodology outperformed the benchmark with an average score of 0.4185, 49.50, and 49.16 for cosine similarity, exact match, and partial match, respectively.

BackgroundPimavanserin, a serotonin 5HT-2A receptor inverse agonist is the first-line, FDA-approved treatment of hallucinations and delusions associated with Parkinson’s Disease psychosis (PDP), which occurs in up to 50% of PD patients. The neurobiological mechanism underlying the therapeutic effectiveness of Pimavanserin in PDP remains unknown. Several earlier studies have shown that treatment with 5HT-2A antagonists and other drugs acting on the serotonergic system such as SSRIs increase Brain derived neurotrophic factor (BDNF) levels in rodents. BDNF is synthesized as the precursor proBDNF, that undergoes cleavage intra or extracellularly to produce a mature BDNF (mBDNF) protein. mBDNF is believed to play a key role in neuroplasticity and neurogenesis. The present study tested the hypothesis that treatment with Pimavanserin is associated with higher and sustained elevations of mBDNF.MethodsAdult Sprague–Dawley male rats were treated with Pimavanserin, Fluoxetine or vehicle for 4 weeks (chronic) or 2 h (acute). BDNF levels were determined by enzyme-linked Immunosorbent assay (ELISA).ResultsWe found significant increases in plasma mBDNF levels in rats following chronic Pimavanserin treatment, but not in Fluoxetine-treated rats. No significant changes in mBDNF levels were found in the prefrontal cortex or hippocampus following Pimavanserin or Fluoxetine treatment.ConclusionThese findings suggest that increase in mBDNF levels could be a contributing mechanism for the neuroprotective potential of Pimavanserin.

Iron line spectroscopy has been one of the leading methods not only for measuring the spins of accreting black holes but also for testing fundamental physics. Basing on such a method, we present an analysis of a data set observed simultaneously by NuSTAR and NICER for the black hole binary candidate MAXI J1803-298, which shows prominent relativistic reflection features. Various relxill _ nk flavors are utilized to test the Kerr black hole hypothesis. The results obtained from our analysis provide stringent constraints on Johannsen deformation parameter α _13 with the highest precise to date, namely ${\alpha }_{13}={0.023}_{-0.038}^{+0.071}$ from relxillD _ nk and ${\alpha }_{13}={0.006}_{-0.022}^{+0.045}$ from relxillion _ nk, respectively, in 3 σ credible lever, where the Johannsen metric reduces to the Kerr metric when α _13 vanishes. Furthermore, we investigate the best model fit results using Akaike information criterion and assess its systematic uncertainties.

Abstract Increased levels of pro-inflammatory cytokines have been reported in postmortem brain samples and in the blood of depressed subjects. However, the inflammatory pathways that lead to depressive-like symptoms are not well understood. Using the learned helplessness (LH) model of depression, we examined the role of C1q, the initiator of classical complement pathway in mediating stress-induced depressive-like behavior in mice. We observed no significant changes in social behavior, despair behavior, spatial memory, and aggressive behavior between the wild type (WT) and C1q knockout (KO) mice. However, C1q deletion exacerbated the inescapable electric foot shock-induced learned helplessness behavior in mice. We found significant reductions in C1q mRNA levels in the prefrontal cortex (PFC) of WT helpless mice as compared to the naïve mice. Increased levels of pro-inflammatory cytokines were found in the PFC of C1q KO mice. These findings suggest that classical complement pathway-mediated learned helplessness behavior is accompanied by neuroinflammatory changes under stressful conditions.

Complement system is one of the most important defense mechanisms of the innate immune system. In addition to their roles in immune regulation, complement proteins are also involved in neurodevelopment and adult brain plasticity. Complement dysregulation has been shown in neurodevelopmental disorders including schizophrenia and autism spectrum disorder as well as in mood disorders. A number of clinical as well as genetic studies suggest the role of complement proteins in the cortical thinning and excessive synaptic pruning frequently associated with schizophrenia. The changes in complement proteins are also associated with the pathophysiology of autism spectrum disorder, major depressive disorder and bipolar disorder, but warrant further research. In addition, rodent models suggest a strong case for complement system in anxiety-like behavior. In this article, we review the recent findings on the role of complement system in neuropsychiatric disorders. The possible uses for future complement targeted therapies are also discussed.

Iron-sulfur (Fe-S) cluster proteins carry out essential cellular functions in diverse organisms, including the human pathogen Mycobacterium tuberculosis (Mtb). The mechanisms underlying Fe-S cluster biogenesis are poorly defined in Mtb. Here, we show that Mtb SufT (Rv1466), a DUF59 domain-containing essential protein, is required for the Fe-S cluster maturation. Mtb SufT homodimerizes and interacts with Fe-S cluster biogenesis proteins; SufS and SufU. SufT also interacts with the 4Fe-4S cluster containing proteins; aconitase and SufR. Importantly, a hyperactive cysteine in the DUF59 domain mediates interaction of SufT with SufS, SufU, aconitase, and SufR. We efficiently repressed the expression of SufT to generate a SufT knock-down strain in Mtb (SufT-KD) using CRISPR interference. Depleting SufT reduces aconitase's enzymatic activity under standard growth conditions and in response to oxidative stress and iron limitation. The SufT-KD strain exhibited defective growth and an altered pool of tricarboxylic acid cycle intermediates, amino acids, and sulfur metabolites. Using Seahorse Extracellular Flux analyzer, we demonstrated that SufT depletion diminishes glycolytic rate and oxidative phosphorylation in Mtb. The SufT-KD strain showed defective survival upon exposure to oxidative stress and nitric oxide. Lastly, SufT depletion reduced the survival of Mtb in macrophages and attenuated the ability of Mtb to persist in mice. Altogether, SufT assists in Fe-S cluster maturation and couples this process to bioenergetics of Mtb for survival under low and high demand for Fe-S clusters.

Abstract Kaluza–Klein theory is a popular alternative theory of gravity, with both non-rotating and rotating black hole solutions known. This allows for the possibility that the theory could be observationally tested. We present a model which calculates the reflection spectrum of a black hole accretion disk system, where the black hole is described by a rotating solution of the Kaluza–Klein theory. We also use this model to analyze X-ray data from the stella-mass black hole in GRS 1915+105 and provide constraints on the free parameters of the Kaluza–Klein black holes.

Abstract We construct a relativistic reflection model in a non-Kerr spacetime in which, depending on the value of the deformation parameter of the metric, there are black hole solutions with spin parameter $$|a_*| > 1$$ |a∗|>1 . We apply our model to fit Suzaku data of four Seyfert galaxies (Ton S180, Ark 120, 1H0419–577, and Swift J0501.9–3239). These galaxies host at the center supermassive black holes that were previously interpreted as near-extremal Kerr black holes. For Ton S180 and 1H0419–577, our measurements are still consistent with the Kerr hypothesis. For Ark 120 and Swift J0501.9–3239, the Kerr solution is not recovered at 3-$$\sigma $$ σ . We discuss our results and possible systematic uncertainties in the model.

Summary: Tuberculosis (TB) treatment involves a multidrug regimen for six months, and until two months, it is unclear if treatment is effective. This delay can lead to the evolution of drug resistance, lung damage, disease spread, and transmission. We identify a blood-based 9-gene signature using a computational pipeline that constructs and interrogates a genome-wide transcriptome-integrated protein-interaction network. The identified signature is able to determine treatment response at week 1–2 in three independent public datasets. Signature-based R9-score correctly detected treatment response at individual timepoints (204 samples) from a newly developed South Indian longitudinal cohort involving 32 patients with pulmonary TB. These results are consistent with conventional clinical metrics and can discriminate good from poor treatment responders at week 2 (AUC 0.93(0.81–1.00)). In this work, we provide proof of concept that the R9-score can determine treatment effectiveness, making a case for designing a larger clinical study.

Rapid urbanization and population growth are stressing the present agricultural systems and could threaten food security in the near future. Sustainable development in agriculture is a way out to such enormously growing food demand. Plant growth-promoting bacteria (PGPB) are considered pivotal to providing adequate nutrition and health to plants and maintaining soil microbial dynamics. In the present study, municipal solid waste composts (MSWC) were studied for the presence of PGPB and their growth-promoting characteristics such as ammonium production, siderophores production, phosphorus solubilization and potassium solubilization, IAA (indole acetic acid), and HCN production. Four promising isolates were chosen and identified through 16S rRNA sequencing as Bacillus sp. strain L5-1, Bacillus pumilus strain EE107-P5, Bacillus sp. strain LSRBMoFPIKRGCFTRI6 and Bacillus sp. strain LPOC3. The potential of isolates is validated using Solanum lycopersicum (tomato) and was found to improve its growth significantly. The findings indicated the presence of potential Bacillus strains in MSWCs, and these composts can be utilized as biofertilizers for urban agricultural practices. However, studies concerning their impact on other crops’ growth and health are still underdeveloped. Since MSWCs might carry hazardous metals or chemicals, their evaluation for the safe application on the crops should also be assessed.

Climate change is more likely to have a detrimental effect on the world’s productive assets. Several undesirable conditions and practices, including extreme temperature, drought, and uncontrolled use of agrochemicals, result in stresses that strain agriculture. In addition, nutritional inadequacies in food crops are wreaking havoc on human health, especially in rural regions of less developed countries. This could be because plants are unable to absorb the nutrients in conventional fertilizers, or these fertilizers have an inappropriate or unbalanced nutrient composition. Chemical fertilizers have been used for centuries and have considerably increased crop yields. However, they also disrupt soil quality and structure, eventually impacting the entire ecosystem. To address the situation, it is necessary to develop advanced materials that can release nutrients to targeted points in the plant-soil environment or appropriate receptors on the leaf in the case of foliar applications. Recently, nanotechnology-based interventions have been strongly encouraged to meet the world’s growing food demand and to promote food security in an environmentally friendly manner. Biological approaches for the synthesis of nanoscale agro-materials have become a promising area of research, with a wide range of product types such as nanopesticides, nanoinsecticides, nanoherbicides, nanobactericides/fungicides, bio-conjugated nanocomplexes, and nanoemulsions emerging therefrom. These materials are more sustainable and target-oriented than conventional agrochemicals. In this paper, we reviewed the literature on major abiotic and biotic stresses that are detrimental to plant growth and productivity. We comprehensively discussed the different forms of nanoscale agro-materials and provided an overview of biological approaches in nano-enabled strategies that can efficiently alleviate plant biotic and abiotic stresses while potentially enhancing the nutritional values of plants.

The ultraluminous X-ray source CXO J133815.6+043255 is a strong candidate for a bona fide intermediate-mass black hole residing in the outskirts of NGC 5252. We present 22 GHz radio observations of this source obtained serendipitously in an ongoing high-frequency imaging survey of radio-quiet Active Galactic Nuclei (AGNs), and use this new data point to construct the broadband radio spectral energy distribution (SED). We find that the SED exhibits a spectral slope of α = −0.66 ± 0.02, consistent with a steep spectrum from optically thin synchrotron emission from an unresolved jet. We also find that the L _R / L _X ratio is approximately 10 ^−3 , inconsistent with radio-quiet AGN and many ULXs but consistent with low-luminosity AGN and radio-loud quasars. Together, these observations support the conclusion that CXO J133815.6+043255 is an intermediate-mass black hole producing a low-mass analog of radio jets seen in classical quasars.

We perform correlation and periodicity search analyses on long-term multiband light curves of the flat-spectrum radio quasar PKS 1510−089 observed by the space-based Fermi-Large Area Telescope in γ -rays, the SMARTS and Steward Observatory telescopes in optical and near-infrared (NIR), and the 13.7 m radio telescope in Metsähovi Radio Observatory between 2008 and 2018. The z -transform discrete correlation function method is applied to study the correlation and possible time lags among these multiband light curves. Among all pairs of wavelengths, the γ -ray versus optical/NIR and optical versus NIR correlations show zero time lags; however, both the γ -ray and optical/NIR emissions precede the radio radiation. The generalized Lomb–Scargle periodogram, weighted wavelet z -transform, and REDFIT techniques are employed to investigate the unresolved core emission–dominated 37 GHz light curve and yield evidence for a quasi period around 1540 days, although given the length of the whole data set it cannot be claimed to be significant. We also investigate the optical/NIR color variability and find that this source shows a simple redder-when-brighter behavior over time, even in the low-flux state.

The persistence of Mycobacterium tuberculosis (Mtb) is a major problem in managing tuberculosis (TB). Host-generated nitric oxide (NO) is perceived as one of the signals by Mtb to reprogram metabolism and respiration for persistence. However, the mechanisms involved in NO sensing and reorganizing Mtb's physiology are not fully understood. Since NO damages iron-sulfur (Fe–S) clusters of essential enzymes, the mechanism(s) involved in regulating Fe–S cluster biogenesis could help Mtb persist in host tissues. Here, we show that a transcription factor SufR (Rv1460) senses NO via its 4Fe–4S cluster and promotes persistence of Mtb by mobilizing the Fe–S cluster biogenesis system; suf operon (Rv1460-Rv1466). Analysis of anaerobically purified SufR by UV–visible spectroscopy, circular dichroism, and iron-sulfide estimation confirms the presence of a 4Fe–4S cluster. Atmospheric O2 and H2O2 gradually degrade the 4Fe–4S cluster of SufR. Furthermore, electron paramagnetic resonance (EPR) analysis demonstrates that NO directly targets SufR 4Fe–4S cluster by forming a protein-bound dinitrosyl-iron-dithiol complex. DNase I footprinting, gel-shift, and in vitro transcription assays confirm that SufR directly regulates the expression of the suf operon in response to NO. Consistent with this, RNA-sequencing of MtbΔsufR demonstrates deregulation of the suf operon under NO stress. Strikingly, NO inflicted irreversible damage upon Fe–S clusters to exhaust respiratory and redox buffering capacity of MtbΔsufR. Lastly, MtbΔsufR failed to recover from a NO-induced non-growing state and displayed persistence defect inside immune-activated macrophages and murine lungs in a NO-dependent manner. Data suggest that SufR is a sensor of NO that supports persistence by reprogramming Fe–S cluster metabolism and bioenergetics.

The advent of combined antiretroviral treatment (cART) as a treatment for HIV-1 infection has not only resulted in a dramatic decrease in the peripheral viral load but has also led to increased life expectancy of the infected individuals. Paradoxically, increased lifespan is accompanied with higher prevalence of age-related comorbidities, including HIV-associated neurocognitive disorders (HAND). Present study was aimed at exploring the role of HIV TAT protein in mediating microglial mitochondrial oxidative stress, ultimately resulting in neuroinflammation and microglial senescence. Our findings demonstrated that exposure of mouse primary microglial cells (mPMs) to HIV TAT protein resulted in a senescence-like phenotype, that was characterized by elevated expression of both p16 and p21 proteins, increased numbers of senescence-associated-β-galactosidase positive cells, augmented cell-cycle arrest, increased release of proinflammatory cytokines and decreased telomerase activity. Additionally, exposure of mPMs to HIV TAT also resulted downregulation of SIRT3 with a concomitant increase in mitochondrial oxidative stress. Dual luciferase reporter assay identified miR-505 as a novel target of SIRT3, which was upregulated in mPMs exposed to HIV TAT. Furthermore, transient transfection of mPMs with either the SIRT3 plasmid or miRNA-505 inhibitor upregulated the expression of SIRT3 and mitochondrial antioxidant enzymes, with a concomitant decrease in microglial senescence. These in vitro findings were also validated in the prefrontal cortices and striatum of HIV transgenic rats as well as cART-treated HIV-infected individuals. In summary, this study underscores a yet undiscovered novel mechanism(s) underlying HIV TAT-mediated induction of senescence phenotype in microglia, involving the miR-505-SIRT3 axis-mediated induction of mitochondrial oxidative stress.

Successful suppression of viral replication by combined antiretroviral therapy (cART) in HIV-1 infected individuals is paradoxically also accompanied by an increased prevalence of HIV-associated neurocognitive disorders (HAND) in these individuals. HAND is characterized by a state of chronic oxidative stress and inflammation. Microglia are extremely sensitive to a plethora of stimuli, including viral proteins and cART. The current study aimed to assess the effects of cART-mediated oxidative stress on the induction of inflammatory responses in microglia. In the present study, we chose a combination of three commonly used antiretroviral drugs—tenofovir disoproxil fumarate, emtricitabine, and dolutegravir. We demonstrated that exposure of microglia to the chosen cART cocktail induced generation of reactive oxygen species, subsequently leading to lysosomal dysfunction and dysregulated autophagy, ultimately resulting in the activation of microglia. Intriguingly, the potent antioxidant, N-acetylcysteine, reversed the damaging effects of cART. These in vitro findings were further corroborated in vivo wherein cART-treated HIV transgenic (Tg) rats demonstrated increased microglial activation, exaggerated lysosome impairment, and dysregulated autophagy in the prefrontal cortices compared with HIV Tg rats not exposed to cART. Similar to in vitro findings, the treatment of HIV Tg rats with N-acetylcysteine also mitigated the deleterious effects of cART. Taken together, our findings suggest that oxidative stress-mediated lysosomal dysfunction plays a critical role in the pathogenesis of HAND in drug-treated HIV-infected individuals and that antioxidant-mediated mitigation of oxidative stress could thus be considered as an adjunctive therapeutic strategy for ameliorating/dampening some of the neurological complications of HAND.

Yeast Sfh5 is an unusual member of the Sec14-like phosphatidylinositol transfer protein (PITP) family. Whereas PITPs are defined by their abilities to transfer phosphatidylinositol between membranes in vitro, and to stimulate phosphoinositide signaling in vivo, Sfh5 does not exhibit these activities. Rather, Sfh5 is a redox-active penta-coordinate high spin FeIII hemoprotein with an unusual heme-binding arrangement that involves a co-axial tyrosine/histidine coordination strategy and a complex electronic structure connecting the open shell iron d-orbitals with three aromatic ring systems. That Sfh5 is not a PITP is supported by demonstrations that heme is not a readily exchangeable ligand, and that phosphatidylinositol-exchange activity is resuscitated in heme binding-deficient Sfh5 mutants. The collective data identify Sfh5 as the prototype of a new class of fungal hemoproteins, and emphasize the versatility of the Sec14-fold as scaffold for translating the binding of chemically distinct ligands to the control of diverse sets of cellular activities.

MicroRNA-124 (miR-124), a brain-enriched microRNA, is known to regulate microglial quiescence. Psychostimulants such as cocaine have been shown to activate microglia by downregulating miR-124, leading, in turn, to neuroinflammation. We thus rationalized that restoring the levels of miR-124 could function as a potential therapeutic approach for cocaine-mediated neuroinflammation. Delivering miRNA based drugs in the brain that are effective and less invasive, however, remains a major challenge in the field. Herein we engineered extracellular vesicles (EVs) and loaded them with miR-124 for delivery in the brain. Approach involved co-transfection of mouse dendritic cells with Dicer siRNA and RVG-Lamp2b plasmid to deplete endogenous miRNAs and for targeting the CNS, respectively. Mouse primary microglia (mPm) were treated with purified engineered EVs loaded with either Cy5-miR-124 or Cy5-scrambled miRNA oligos in the presence or absence of cocaine followed by assessing EV uptake and microglial activation. In vivo studies involved pretreating mice intranasally with engineered EVs followed by cocaine injection (20 mg/kg, i.p.). mPm exposed to EV-miR-124 exhibited reduced expression of miR-124 targets – TLR4 and STAT3 as well as ERK-1/2 and Iba1. In cocaine administered mice, EV-Cy5-miR-124 delivered intranasally were detected in the CNS and significantly reduced the expression of inflammatory markers TLR4, MYD88, STAT3 and NF-kB p65 while also downregulating the microglial activation marker, Iba1. Collectively, these findings suggest that engineered EVs can deliver miR-124 into the CNS, thereby alleviating cocaine-mediated microglial activation. Manipulating EV miRNAs can thus be envisioned as an efficient means for delivery of RNA-based therapeutics to target organs.

Sec14-like phosphatidylinositol transfer proteins (PITPs) play important biological functions in integrating multiple aspects of intracellular lipid metabolism with phosphatidylinositol-4-phosphate signaling. As such, these proteins offer new opportunities for highly selective chemical interference with specific phosphoinositide pathways in cells. The first and best characterized small molecule inhibitors of the yeast PITP, Sec14, are nitrophenyl(4-(2-methoxyphenyl)piperazin-1-yl)methanones (NPPMs), and a hallmark feature of NPPMs is their exquisite targeting specificities for Sec14 relative to other closely related Sec14-like PITPs. Our present understanding of Sec14::NPPM binding interactions is based on computational docking and rational loss-of-function approaches. While those approaches have been informative, we still lack an adequate understanding of the basis for the high selectivity of NPPMs among closely related Sec14-like PITPs. Herein, we describe a Sec14 motif, which we term the VV signature, that contributes significantly to the NPPM sensitivity/resistance of Sec14-like phosphatidylinositol (PtdIns)/phosphatidylcholine (PtdCho) transfer proteins. The data not only reveal previously unappreciated determinants that govern Sec14-like PITP sensitivities to NPPMs, but enable predictions of which Sec14-like PtdIns/PtdCho transfer proteins are likely to be NPPM resistant or sensitive based on primary sequence considerations. Finally, the data provide independent evidence in support of previous studies highlighting the importance of Sec14 residue Ser173 in the mechanism by which NPPMs engage and inhibit Sec14-like PITPs.

Acetaminophen (APAP) is a commonly used analgesic and antipyretic that can cause hepatotoxicity due to production of toxic metabolites via cytochrome P450 (Cyp) 1a2 and Cyp2e1. Previous studies have shown conflicting effects of fructose (the major component in Western diet) on the susceptibility to APAP-induced hepatotoxicity. To evaluate the role of fructose-supplemented diet in modulating the extent of APAP-induced liver injury, male C57BL/6J mice were given 30% (w/v) fructose in water (or regular water) for 8 weeks, followed by oral administration of APAP. APAP-induced liver injury (determined by serum levels of liver enzymes) was decreased by two-fold in mice pretreated with fructose. Fructose-treated mice exhibited (~1.5 fold) higher basal glutathione levels and (~2 fold) lower basal (mRNA and activity) levels of Cyp1a2 and Cyp2e1, suggesting decreased bioactivation of APAP and increased detoxification of toxic metabolite in fructose-fed mice. Hepatic mRNA expression of heat shock protein 70 was also found increased in fructose-fed mice. Analysis of bacterial 16S rRNA gene amplicons from the cecal samples of vehicle groups showed that the fructose diet altered gut bacterial community, leading to increased α-diversity. The abundance of several bacterial taxa including the genus Anaerostipes was found to be significantly correlated with the levels of hepatic Cyp2e1, Cyp1a2 mRNA, and glutathione. Together, these results suggest that the fructose-supplemented diet decreases APAP-induced liver injury in mice, in part by reducing metabolic activation of APAP and inducing detoxification of toxic metabolites, potentially through altered composition of gut microbiota.

In the era of combined antiretroviral therapy (cART), as infected individuals continue to have longer lifespans, there is also an increased prevalence of HIV-associated neurocognitive disorders (HAND). Inflammation is one of the underlying features of HAND, with the role of viral proteins and antiretroviral drugs implicated in this process. Microglia are extremely sensitive to a plethora of stimuli, including viral products and cART. The current study was undertaken to understand the molecular mechanism(s) underlying cART-mediated activation of microglia. Herein we chose a combination of three commonly used drugs, tenofovir disoproxil fumarate (TDF), emtricitabine (FTC), and dolutegravir (DTG). We demonstrated that exposure of microglia to this cART cocktail induced lysosomal membrane permeabilization (LMP), which subsequently resulted in impaired lysosomal functioning involving elevated pH and decreased cathepsin D (CTSD) activity. cART exposure of microglia resulted in increased formation of autophagosomes as demonstrated by a time-dependent increase of autophagy markers, with a concomitant defect in the fusion of the lysosomes with the autophagosome. Taken together, our findings suggest a novel mechanism by which cART impairs lysosomal functioning, resulting in dysregulated autophagy and increased neuroinflammation. Interventions aimed at lysosome protection could likely be envisioned as promising therapeutic targets for abrogating cART-mediated microglia activation, which in turn, could thus be considered as adjunctive therapeutics for the treatment of HAND pathogenesis.

In this paper, we review our current efforts to test General Relativity in the strong field regime by studying the reflection spectrum of supermassive black holes. So far we have analyzed 11 sources with observations of NuSTAR, Suzaku, Swift, and XMM-Newton. Our results are consistent with General Relativity, according to which the spacetime metric around astrophysical black holes should be well approximated by the Kerr solution. We discuss the systematic uncertainties in our model and we present a preliminary study on the impact of some of them on the measurement of the spacetime metric.

Background. Chronic obstructive pulmonary disease (COPD) is a common, smoking-related lung disease. Patients with COPD frequently suffer disease exacerbations induced by bacterial respiratory infections, suggestive of impaired innate immunity. Low-dose oxygen is a mainstay of therapy during COPD exacerbations; yet we understand little about whether oxygen can modulate the effects of cigarette smoke on lung immunity. Methods. Wild-type mice were exposed to cigarette smoke for 5 weeks, followed by intratracheal instillation of Pseudomonas aeruginosa (PAO1) and 21% or 35–40% oxygen. After two days, lungs were harvested for PAO1 CFUs, and bronchoalveolar fluid was sampled for inflammatory markers. In culture, macrophages were exposed to cigarette smoke and oxygen (40%) for 24 hours and then incubated with PAO1, followed by quantification of bacterial phagocytosis and inflammatory markers. Results. Mice exposed to 35–40% oxygen after cigarette smoke and PAO1 had improved survival and reduced lung CFUs and inflammation. Macrophages from these mice expressed less TNF-α and more scavenger receptors. In culture, macrophages exposed to cigarette smoke and oxygen also demonstrated decreased TNF-α secretion and enhanced phagocytosis of PAO1 bacteria. Conclusions. Our findings demonstrate a novel, protective role for low-dose oxygen following cigarette smoke and bacteria exposure that may be mediated by enhanced macrophage phagocytosis.

Einstein’s theory of general relativity was proposed over 100 years ago and has successfully passed a large number of observational tests in the weak field regime. However, the strong field regime is largely unexplored, and there are many modified and alternative theories that have the same predictions as Einstein’s gravity for weak fields and present deviations when gravity becomes strong. relxill_nk is the first relativistic reflection model for probing the spacetime metric in the vicinity of astrophysical black holes and testing Einstein’s gravity in the strong field regime. Here, we present our current constraints on possible deviations from Einstein’s gravity obtained from the black holes in 1H0707–495, Ark 564, GX 339–4, and GS 1354–645.

Introduction: Management of type 2 diabetes mellitus (DM) needs multi factorial risk reduction strategies like weight reduction, blood pressure (BP) control and lipid profile regulation, along with glycaemic control. These strategies should be implemented very early in the course of the disease to prevent both microvascular and macro vascular complications. Aim: To find out the prevalence of diabetic complications and to audit the management of glycaemia, blood pressure and serum lipids in the outpatient diabetes clinic in a hospital located in Western U.P. Materials and Methods: A prospective study conducted in a tertiary referral teaching hospital, on 105 patients with type 2 diabetes mellitus (mean age 52.84 ± 1.02 years) who attended the diabetic clinic during June 2014 to April 2015, who had undergone screening for complications. Main outcome measures: Weight, height, body mass index (BMI), blood pressure, fasting and post prandial blood sugars, glycated hemoglobin, serum lipids and presence of diabetic microvascular as well as macrovascular complications. Mean of various clinical and biochemical parameters were compared in patients with and without complications. Results: The mean age of patients was estimated as 52.84 ± 1.02 years. Fifty six percent of patients were males. Average BMI was 28.61 ± 0.28 kg/m2 . Most of them had abdominal obesity. The average duration of diabetes was 8.39 ± 0.6 years. The average of systolic blood pressure (SBP), diastolic blood pressure (DBP), HbA1C, FBS, PPBS, LDL, HDL and triglyceride were estimated as 129.1 ± 1.49 mm Hg, 84.85 ± 0.94 mm Hg, 6.99 ± 0.08%, 141.33 ± 2.12 mg/dl, 214.51 ± 3.11 mg/dl, 155.66 ± 2.07 mg /dl, 40.07 ± 0.38 mg/dl, 236.53 ± 3.31 mg/dl respectively based on 105 patients. Amongst micro vascular complications; retinopathy, neuropathy and nephropathy were documented in 51.4, 77.14 and 30.47% diabetes patients respectively. The prevalence of coronary artery disease, stroke and gangrene were 42.85, 12.38 and 3.80% respectively. Conclusion: The prevalence and risk of the complications of type 2 DM significantly increased with age of patients, duration of diabetes, fasting blood sugar and LDL levels. The awareness regarding monitoring of target BP and lipid profile is needed beyond the glycaemic control, amongst patients and health care providers.