RNA-Seq time-course analysis of neural precursor cell transcriptome in response to herpes simplex Virus-1 infection.

The neurogenic niches within the central nervous system serve as essential reservoirs for neural precursor cells (NPCs), playing a crucial role in neurogenesis. However, these NPCs are particularly vulnerable to infection by the herpes simplex virus 1 (HSV-1). In the present study, we investigated the changes in the transcriptome of NPCs in response to HSV-1 infection using bulk RNA-Seq, compared to those of uninfected samples, at different time points post infection and in the presence or absence of antivirals. The results showed that NPCs upon HSV-1 infection undergo a significant dysregulation of genes playing a crucial role in aspects of neurogenesis, including genes affecting NPC proliferation, migration, and differentiation. Our analysis revealed that the CREB signaling, which plays a crucial role in the regulation of neurogenesis and memory consolidation, was the most consistantly downregulated pathway, even in the presence of antivirals. Additionally, cholesterol biosynthesis was significantly downregulated in HSV-1-infected NPCs. The findings from this study, for the first time, offer insights into the intricate molecular mechanisms that underlie the neurogenesis impairment associated with HSV-1 infection.

Environmentally relevant lead alters nuclear integrity in erythrocytes and generates oxidative stress in liver of Anabas testudineus: Involvement of Nrf2-Keap1 regulation and expression of biomarker genes.

Genotoxic and hepatotoxic effects of lead (Pb) on a freshwater fish, climbing perch (Anabas testudineus) were studied at an environmentally relevant concentration (43.3 ppm). The genotoxic potential of Pb was confirmed by micronucleus study, with increased frequencies of erythrocytic nuclear alterations like lobed, blebbed, notched, fragmented, and micronuclei were observed in erythrocytes in treated groups as compared to control. Inorganic Pb induces oxidative stress which is a consequence of elevated level of Reactive Oxygen Species. Hepatotoxicity was assessed both by the oxidative stress and cellular responses that emerged due to the toxic assault of Pb in the liver, the most important detoxifying organ. Upregulation of xenobiotic metabolizing enzyme like catalase was evident after 15, 30, and 90 days of exposure, and a profound effect was observed on 30th days. The level of lipid peroxidation and reduced glutathione was increased after Pb exposure. Histoarchitectural damages of liver were distinctly evident in treated fish. Western blot analysis confirmed the expressional alterations of stress-responsive marker proteins like Nrf2, Keap1, Hsp70, and Nqo1. Pb exposure resulted in increased expression of Hsp70, Nrf2, and Nqo1, whereas Keap1 was downregulated, suggesting the involvement of Nrf2-Keap1 regulation as a cytoprotective mechanism against Pb toxicity.

Chronic low-dose chromium VI exposure induces oxidative stress and apoptosis with altered expressions of DNA repair genes and promoter hypermethylation in the liver of Swiss albino mice.

The present investigation dealt with harmful effects of hexavalent chromium (Cr [VI]) on liver of Swiss albino mice. This variant exhibited cytotoxicity, mutagenicity, and carcinogenicity. Our study focused on elucidating the hepatotoxic effects of chronic low-dose exposure to Cr (VI) (2, 5, and 10 ppm) administered via drinking water for 4 and 8 months. The observed elevation in SGPT, ALP, and SGOT and increased oxidative stress markers unequivocally confirmed the severe disruption of liver homeostasis at these low treatment doses. Noteworthy alterations in histoarchitecture, body weight, and water intake provided further evidences of the harmful effects of Cr (VI). Production of reactive oxygen species (ROS) during metabolism led to DNA damages. Immunohistochemistry and qRT-PCR analyses revealed that chronic low-dose exposure of Cr (VI) induced apoptosis in liver tissue. Our study exhibited alterations in the expression pattern of DNA repair genes (Rad51, Mutyh, Mlh1, and Ogg1), coupled with promoter hypermethylation of Mutyh and Rad51, leading to transcriptional inhibition. Our findings underscored the potential of low-dose Cr (VI) exposure on hepatotoxicity by the intricate interplay between apoptosis induction and epigenetic alterations of DNA repair genes.

Differential detection of aspartic acid in MCF-7 breast cancer cells.

Aspartic acid is a non-essential amino acid obtained in the neuroendocrine tissues of vertebrates and invertebrates. Aspartic acid, a major excitatory neurotransmitter in the mammalian central nervous system, plays a key role in memory and acts in many other normal and abnormal physiological processes. In this work, we have developed an efficient chemosensor (PCF) based on the pyridine-carbazole moiety for the differential detection of aspartic acid in biological systems. PCF has a strong binding affinity towards aspartic acid, with a detection limit in the nanomolar range. The binding stoichiometry of aspartic aid and PCF was obtained as 1 : 1 from a Jobs plot analysis. Furthermore, the efficacy of PCF has been successfully demonstrated in in vitro experiments in MCF-7 breast cancer cells.

Direct fluorescence labelling of NO inside plant cells.

Nitric oxide (NO) plays a key role in regulating plant growth, enhances nutrient uptake, and activates disease and stress tolerance mechanisms in most plants. NO is marked as a potential tool for improving the yield and quality of horticultural crop species. Research on NO in plant species can provide an abundance of valuable information regarding this. Hence, we have prepared a simple chemosensor (NPO) for the detection of endogenous NO in chickpea saplings. NPO selectively interacts with NO as determined through a chemodosimetric method to clearly show both the colorimetric and fluorometric changes. After the interaction with NO, the colorless NPO turns yellow as observed by the naked eye and shows bright cyan-blue fluorescence under a UV lamp. The 1 : 1 stoichiometric ratio between NPO and NO is determined from Job's plot resulting in a stable diazeniumdiolate product. The interaction mechanism is well established by absorption, fluorescence titration, NMR titration, HRMS, and DFT calculations. This method has successfully been employed in the plant's root and stem systems to label NO. Confocal microscopy images might help us to understand the endogenous NO generation and the mechanism that happens inside plant tissues.

Chronic exposure to environmentally relevant concentration of fluoride impairs osteoblast's collagen synthesis and matrix mineralization: Involvement of epigenetic regulation in skeletal fluorosis.

Globally, 200 million people are suffering from toxic manifestations of Fluoride(F), dental and skeletal fluorosis; unfortunately, there is no treatment. To unravel the pathogenesis of skeletal fluorosis, we established fluorosis mice by treating environmentally relevant concentration of F (15 ppm NaF) through drinking water for 4 months. As in skeletal fluorosis, locomotor disability, crippling deformities occur and thus, our hypothesis was F might adversely affects collagen which gives the bone tensile strength. This work inevitably had to be carried out on osteoblast cells, responsible for synthesis, deposition, and mineralization of bone matrix. Isolated osteoblast cells were confirmed by ALP activity and mineralized nodules formation. Expression of collagen Col1a1, Col1a2, COL1A1 was significantly reduced in treated mice. Further, a study revealed the involvement of epigenetic regulation by promoter hypermethylation of Col1a1; expressional alterations of transcription factors, calcium channels and other genes e.g., Cbfa-1, Tgf-ß1, Bmp1, Sp1, Sp7, Nf-Kb p65, Bmp-2, Bglap, Gprc6a and Cav1.2 are associated with impairment of collagen synthesis, deposition and decreased mineralization thus, enfeebling bone health. This study indicates the possible association of epigenetic regulation in skeletal fluorosis. However, no association was found between polymorphisms in the Col1a1 (RsaI, HindIII) and Col1a2 (RsaI, HindIII) genes with fluorosis in mice.

NF-κB Signaling-Mediated Activation of WNK-SPAK-NKCC1 Cascade in Worsened Stroke Outcomes of Ang II-Hypertensive Mice.

BACKGROUND: Worsened stroke outcomes with hypertension comorbidity are insensitive to blood pressure-lowering therapies. In an experimental stroke model with comorbid hypertension, we investigated causal roles of ang II (angiotensin II)-mediated stimulation of the brain WNK (with no lysine [K] kinases)-SPAK (STE20/SPS1-related proline/alanine-rich kinase)-NKCC1 (Na-K-Cl cotransporter) complex in worsened outcomes. METHODS: Saline- or ang II-infused C57BL/6J male mice underwent stroke induced by permanent occlusion of the distal branches of the middle cerebral artery. Mice were randomly assigned to receive either vehicle dimethyl sulfoxide/PBS (2 mL/kg body weight/day, IP), a novel SPAK inhibitor, 5-chloro-N-(5-chloro-4-((4-chlorophenyl)(cyano)methyl)-2-methylphenyl)-2-hydroxybenzamide (ZT-1a' 5 mg/kg per day, IP) or a NF-κB (nuclear factor-κB) inhibitor TAT-NBD (transactivator of transcription-NEMO-binding domain' 20 mg/kg per day, IP). Activation of brain NF-κB and WNK-SPAK-NKCC1 cascade as well as ischemic stroke outcomes were examined. RESULTS: Stroke triggered a 2- to 5-fold increase of WNK (isoforms 1, 2, 4), SPAK/OSR1 (oxidative stress-responsive kinase 1), and NKCC1 protein in the ang II-infused hypertensive mouse brains at 24 hours after stroke, which was associated with increased nuclear translocation of phospho-NF-κB protein in the cortical neurons (a Pearson correlation r of 0.77, P<0.005). The upregulation of WNK-SPAK-NKCC1 cascade proteins resulted from increased NF-κB recruitment on Wnk1, Wnk2, Wnk4, Spak, and Nkcc1 gene promoters and was attenuated by NF-κB inhibitor TAT-NBD. Poststroke administration of SPAK inhibitor ZT-1a significantly reduced WNK-SPAK-NKCC1 complex activation, brain lesion size, and neurological function deficits in the ang II-hypertensive mice without affecting blood pressure and cerebral blood flow. CONCLUSIONS: The ang II-induced stimulation of NF-κB transcriptional activity upregulates brain WNK-SPAK-NKCC1 cascade and contributes to worsened ischemic stroke outcomes, illustrating the brain WNK-SPAK-NKCC1 complex as a therapeutic target for stroke with comorbid hypertension.

Attenuated cell-cycle division protein 2 and elevated mitotic roles of polo-like kinase 1 characterize deficient myoblast fusion in peripheral arterial disease.

INTRODUCTION: We propose that MuSC-derived myoblasts in PAD have transcriptomic differences that can highlight underlying causes of ischemia-induced myopathy. METHODS: Differentiation capacity among perfused and ischemic human myoblasts was compared. Following next generation sequencing of mRNA, Ingenuity Pathway Analysis (IPA) was performed for canonical pathway enrichment. Live cell imaging and immunofluorescence were performed to determine myocyte fusion index and protein expression based on insights from IPA, specifically concerning cell cycle regulators including cell-division cycle protein 2 (CDC2) and polo-like kinase 1 (PLK1). RESULTS: Ischemic myoblasts formed attenuated myotubes indicative of reduced fusion. Additionally, myoblasts from ischemic segments showed significant differences in canonical pathways associated with PLK1 (upregulated) and G2/M DNA damage checkpoint regulation (downregulated). PLK1 inhibition with BI2536 did not affect cell viability in any group over 24 h but deterred fusion more significantly in PAD myoblasts. Furthermore, PLK1 inhibition reduced the expression of checkpoint protein CDC2 in perfused but not ischemic cells. CONCLUSION: Differentiating myoblasts derived from ischemic muscle have significant differences in gene expression including those essential to DNA-damage checkpoint regulation and cell cycle progress. DNA-damage checkpoint dysregulation may contribute to myopathy in PAD.

Molecular Biology Information Service: an innovative medical library-based bioinformatics support service for biomedical researchers.

Biomedical researchers are increasingly reliant on obtaining bioinformatics training in order to conduct their research. Here we present a model that academic institutions may follow to provide such training for their researchers, based on the Molecular Biology Information Service (MBIS) of the Health Sciences Library System, University of Pittsburgh (Pitt). The MBIS runs a four-facet service with the following goals: (1) identify, procure and implement commercially licensed bioinformatics software, (2) teach hands-on workshops using bioinformatics tools to solve research questions, (3) provide in-person and email consultations on software/databases and (4) maintain a web portal providing overall guidance on the access and use of bioinformatics resources and MBIS-created webtools. This paper describes these facets of MBIS activities from 2006 to 2018, including outcomes from a survey measuring attitudes of Pitt researchers about MBIS service and performance.

Environmentally Relevant Hexavalent Chromium Disrupts Elemental Homeostasis and Induces Apoptosis in Zebrafish Liver.

Although hexavalent chromium Cr [VI] is known as a toxicant in the aquatic environment, its effect in low, environmentally relevant concentration (ERC; 2 mg L-1) is less characterized. Against this backdrop, the effects of Cr [VI] in ERC on zebrafish liver has been investigated in this study. Fluorescence microscopy and gel electrophoresis detected excess DNA damage and cell death via apoptosis in 2 mg L-1 Cr [VI]-treated fish when compared with that of control. Besides, there were transcriptional activations of p53, Bax, Caspase 9 and Caspase 3 genes but downregulation of Bcl2 gene in the treated group, confirming the apoptotic pathway. Energy dispersive X-ray fluorescence (EDXRF) data showed significant (p < 0.05) increase in hepatic content of Cr, selenium, iron, manganese, calcium, sulfur and magnesium but depletion of zinc, copper and cobalt in the treated group. Collectively, the study shows that even a low, ERC of Cr [VI] is toxic to the zebrafish as it elicited marked apoptosis in the hepatocytes and altered the liver elemental profile.

Attenuating vascular stenosis-induced astrogliosis preserves white matter integrity and cognitive function.

BACKGROUND: Chronic cerebral hypoperfusion (CCH) causes white matter damage and cognitive impairment, in which astrogliosis is the major pathology. However, underlying cellular mechanisms are not well defined. Activation of Na+/H+ exchanger-1 (NHE1) in reactive astrocytes causes astrocytic hypertrophy and swelling. In this study, we examined the role of NHE1 protein in astrogliosis, white matter demyelination, and cognitive function in a murine CCH model with bilateral carotid artery stenosis (BCAS). METHODS: Sham, BCAS, or BCAS mice receiving vehicle or a selective NHE1 inhibitor HOE642 were monitored for changes of the regional cerebral blood flow and behavioral performance for 28 days. Ex vivo MRI-DTI was subsequently conducted to detect brain injury and demyelination. Astrogliosis and demyelination were further examined by immunofluorescence staining. Astrocytic transcriptional profiles were analyzed with bulk RNA-sequencing and RT-qPCR. RESULTS: Chronic cerebral blood flow reduction and spatial working memory deficits were detected in the BCAS mice, along with significantly reduced mean fractional anisotropy (FA) values in the corpus callosum, external capsule, and hippocampus in MRI DTI analysis. Compared with the sham control mice, the BCAS mice displayed demyelination and axonal damage and increased GFAP+ astrocytes and Iba1+ microglia. Pharmacological inhibition of NHE1 protein with its inhibitor HOE642 prevented the BCAS-induced gliosis, damage of white matter tracts and hippocampus, and significantly improved cognitive performance. Transcriptome and immunostaining analysis further revealed that NHE1 inhibition specifically attenuated pro-inflammatory pathways and NADPH oxidase activation. CONCLUSION: Our study demonstrates that NHE1 protein is involved in astrogliosis with pro-inflammatory transformation induced by CCH, and its blockade has potentials for reducing astrogliosis, demyelination, and cognitive impairment.

Bacopasaponins with cytotoxic activity against human breast cancer cells in vitro.

Globally, breast cancer is a serious concern that exhibits a persistent rise in its incidence and related mortality even after significant advancement in the field of cancer research. To find an alternative cure for the disease from natural resources we selected Bacopa monniera, a perennial ethnomedicinal plant popularly used for boosting memory and mental health. We isolated four different types of dammarane saponins, namely bacopasaponins C-F (1-4) from the plant and evaluated their toxic effects on two different types of human breast cancer cell lines-a hormone-responsive MCF7 and a triple-negative MDA-MB-231. Interestingly, MTT assay revealed a dose-dependent toxic effect of all four types of bacopasaponins on both of these cell lines, 4 being the most effective with 48 h-inhibitory concentration (IC50) of 32.44 and 30 µM in MCF7 and MDA-MB-231 respectively. Further, 4 caused significant alterations in normal cytomorphology and induction of apoptosis in both of these cell lines after 48 h of treatment. No caspase-8 activity was detected in these cell lines when exposed to 4 for 2, 24, and 48 h; instead, Western blotting analysis confirmed involvement of either caspase-9 (MCF7) or both caspase-9 and caspase-3 (MDA-MB-231) in the process of apoptosis indicating the occurrence of intrinsic mode. Additionally, at comparable effective doses to cancer, bacopasaponins showed much less toxicity in normal human peripheral blood lymphocytes (≥ 85% cell survival). Overall, the findings project bacopasaponin F, a natural constituent of Bacopa monniera, as an efficient and safer alternative for breast cancer therapeutics.

Nrf2-ARE signaling in cellular protection: Mechanism of action and the regulatory mechanisms.

Oxidative stress is the increase in cellular oxidant concentration in comparison to antioxidant titer. Toxic insults and many other diseased conditions are mediated through the formation of such condition. Once the redox equilibrium is disrupted, the cellular antioxidant system functions to bring back the cell to redox homeostasis state. The field players of the cytoprotective machinery are the xenobiotic-metabolizing enzymes that are transcriptionally controlled by upstream regulatory pathways like the Nrf2-ARE pathway and AhR-XRE pathway. The importance of Nrf2 lies in the fact that it is activated by a variety of compounds and has a wide range of inducers including metals, organic toxicants and so forth. The present review article aims to discuss the role of Nrf2 in cellular protection and also intends to illuminate the regulatory mechanisms that control Nrf2 itself. This can add to our knowledge of how the cell reacts and survives against such stressed conditions.

Environmental exposure of arsenic and fluoride and their combined toxicity: A recent update.

Environmental exposure to arsenic (As) and fluoride (F) in the recent year has been increased because of excessive use of naturally contaminated ground water. Surface water is also regularly contaminated with these elements in various industrial areas. Arsenicosis and fluorosis upon individual exposure of As and F are reported in many studies. A syndrome of endemic As poisoning and fluorosis occurs during concurrent exposure of As and F. Previous reports showed synergistic, antagonistic and independent effects of these two compounds, although few recent reports also revealed antagonistic effects after co-exposure. Interaction during intestinal absorption and influence of F on As metabolism might be the cause of antagonism. The synergism/antagonism is thought to depend on the dose and duration of the co-exposure. However, the detailed mechanism is still not fully understood and needs further studies. Removal technologies of As and F from contaminated water is available but removal of such contaminants from food is yet to be developed. Antioxidants are useful to mitigate the toxic effects of As and F. This review focused on the effect of co-exposure, amelioration as well as removal techniques of As and F.

Chronic exposure to environmentally relevant concentration of fluoride alters Ogg1 and Rad51 expressions in mice: Involvement of epigenetic regulation.

Chronic exposure to fluoride (F) beyond the permissible limit (1.5 ppm) is known to cause detrimental health effects by induction of oxidative stress-mediated DNA damage overpowering the DNA repair machinery. In the present study, we assessed F induced oxidative stress through monitoring biochemical parameters and looked into the effect of chronic F exposure on two crucial DNA repair genes Ogg1 and Rad51 having important role against ROS induced DNA damages. To address this issue, we exposed Swiss albino mice to an environmentally relevant concentration of fluoride (15 ppm NaF) for 8 months. Results revealed histoarchitectural damages in liver, brain, kidney and spleen. Depletion of GSH, increase in lipid peroxidation and catalase activity in liver and brain confirmed the generation of oxidative stress. qRT-PCR result showed that expressions of Ogg1 and Rad51 were altered after F exposure in the affected organs. Promoter hypermethylation was associated with the downregulation of Rad51. F-induced DNA damage and the compromised DNA repair machinery triggered intrinsic pathway of apoptosis in liver and brain. The present study indicates the possible association of epigenetic regulation with F induced neurotoxicity.

CCAAT/Enhancer-binding protein ß promotes pathogenesis of EAE.

The CCAAT/Enhancer Binding Protein ß (C/EBPß) transcription factor is activated by multiple inflammatory stimuli, including IL-17 and LPS, and C/EBPß itself regulates numerous genes involved in inflammation. However, the role of C/EBPß in driving autoimmunity is not well understood. Here, we demonstrate that Cebpb-/- mice are resistant to EAE. Cebpb-/- mice exhibited reduced lymphocyte and APC infiltration into CNS following EAE induction. Furthermore, MOG-induced Th17 cytokine production was impaired in draining LN, indicating defects in Th17 cell priming. In vitro Th17 polarization studies indicated that T cell responses are not inherently defective, instead supporting the known roles for C/EBPß in myeloid lineage cell activation as the likely mechanism for defective Th17 priming in vivo. However, we did uncover an unexpected role for C/EBPß in regulating ll23r expression in APCs. ChIP assays confirmed that C/EBPß binds directly to the Il23r gene promoter in dendritic cells and Th17 cells. These data establish C/EBPß as a key driver of autoimmune inflammation in EAE, and propose a novel role for C/EBPß in regulation of IL-23R expression.

Silver Nanoparticles as Antibacterial and Anticancer Materials Against Human Breast, Cervical and Oral Cancer Cells.

Silver nanoparticles contribute a giant share to the realm of modern nanobiotechnology. Their utility as antimicrobial agents is also well documented. Green synthesis of nanoparticle has several advantages over its chemical synthesis. In the present study, Thuja occidentalis leaf extract mediated silver nanoparticles were prepared without using a stabilizing agent and tested for their anticancer and anti-microbial activity. Thuja occidentalis leaf extract mediated silver nanoparticles were prepared under ambient conditions which showed a narrow size distribution within the range of 10­15 nm, with average particle size of 12.7 nm. Interestingly, these nanoparticles exhibited anti-cancer activity against human breast (MCF 7, MDA MB 231) and cervical cancer (HeLa) as well as mouth epidermoid carcinoma (KB) cell lines at a concentration range of 6.25­50 µg/mL. Contrarily, they are compatible with human peripheral blood mononuclear cells and rat hepatocytes. Moreover, their efficient inhibitory effect was witnessed against Bacillus subtilis, Staphylococcus aureus, Listeria monocytogenes, Salmonella typhimurium and Pseudomonas aeruginosa with inhibitory concentration at 5­10 µg/mL. The prepared nanoparticles were highly biocompatible and have strong potential in the development of non-toxic chemotherapy with antibacterial attributes.

Differential modulation of cellular antioxidant status in zebrafish liver and kidney exposed to low dose arsenic trioxide.

Zebrafish were exposed to a nonlethal dose (1/350LC50; 50µg/L) of As2O3 and sampled at 7, 15, 30, 60 and 90 days of treatment. The oxidative stress response was assessed in terms of time-dependent histopathological changes, lipid peroxidation, GSH status, activities of detoxification enzymes and expression of antioxidant genes in liver and kidney. As2O3 treatment enhanced lipid peroxidation except at day 90 in liver and day 30 in kidney. Glutathione depleted significantly in the liver except on day 30; whereas in kidney, it increased initially but thereafter depleted significantly. The liver GST activity was high until day 30, low on day 60 and high on day 90. On the other hand, activity of GST in kidney remained high throughout the exposure. GR activity in liver decreased initially but augmented from 30 days onwards whereas in kidney it remained high until 30 days of exposure. Significant increase in GPx and CAT activities in liver and kidney confirmed oxidative stress in zebrafish which correlated with mRNA expression of antioxidant genes. Upregulation in mRNA level of Cu-Zn Sod in liver and kidney was prominent. Gpx1 upregulation was more conspicuous in kidney as compared to liver while the pattern of Cat expression was almost similar in both the organs. Among the mitochondrial genes, expression of Cox1 was significantly high only after 90 days in liver, while in kidney it enhanced at 7, 30 and 60 days of arsenic exposure. Ucp2 was upregulated in liver after 15 days of exposure but significantly downregulated at day 90; in kidney it remained unchanged at other time points except at day 90. An overall increased expression of Bcl2 further confirmed As2O3 induced oxidative stress in zebrafish liver and kidney. The pattern of mRNA expression of Nrf2 was not uniform and was in accordance to its downstream antioxidant genes. Present findings elucidate that low dose of As2O3 exposure induces a time dependent differential modulation of antioxidant status in liver and kidney of zebrafish in a tissue-specific manner.

Structurally Characterized Zn2+ Selective Ratiometric Fluorescence Probe in 100 % Water for HeLa Cell Imaging: Experimental and Computational Studies.

Fluorescence recognition of Zn2+ in 100% aqueous medium using 2-((1, 3 dihydroxy-2-(hydroxymethyl)propan-2 ylimino) methyl) phenol (SALTM) as ratiometric probe is reported. Moreover, SALTM can discriminate Zn2+ from Cd2+very effectively. The binding constant and detection limit of the probe for Zn2+ is 2.2×10(4) M(-1/2) and 2.79×10(-8) M respectively.Interestingly, corresponding naphthalene derivative(HNTM) having less water solubility fails to be a ratiometric sensor. SALTM can detect intracellular Zn2+ in HeLa cervical cancer cells under fluorescence microscope. Moreover, DFT and TD-DFT studies support experimental findings.

Temporal transcriptional response to latency reversing agents identifies specific factors regulating HIV-1 viral transcriptional switch.

BACKGROUND: Latent HIV-1 reservoirs are identified as one of the major challenges to achieve HIV-1 cure. Currently available strategies are associated with wide variability in outcomes both in patients and CD4(+) T cell models. This underlines the critical need to develop innovative strategies to predict and recognize ways that could result in better reactivation and eventual elimination of latent HIV-1 reservoirs. RESULTS AND DISCUSSION: In this study, we combined genome wide transcriptome datasets post activation with Systems Biology approach (Signaling and Dynamic Regulatory Events Miner, SDREM analyses) to reconstruct a dynamic signaling and regulatory network involved in reactivation mediated by specific activators using a latent cell line. This approach identified several critical regulators for each treatment, which were confirmed in follow-up validation studies using small molecule inhibitors. Results indicate that signaling pathways involving JNK and related factors as predicted by SDREM are essential for virus reactivation by suberoylanilide hydroxamic acid. ERK1/2 and NF-κB pathways have the foremost role in reactivation with prostratin and TNF-α, respectively. JAK-STAT pathway has a central role in HIV-1 transcription. Additional evaluation, using other latent J-Lat cell clones and primary T cell model, also confirmed that many of the cellular factors associated with latency reversing agents are similar, though minor differences are identified. JAK-STAT and NF-κB related pathways are critical for reversal of HIV-1 latency in primary resting T cells. CONCLUSION: These results validate our combinatorial approach to predict the regulatory cellular factors and pathways responsible for HIV-1 reactivation in latent HIV-1 harboring cell line models. JAK-STAT have a role in reversal of latency in all the HIV-1 latency models tested, including primary CD4(+) T cells, with additional cellular pathways such as NF-κB, JNK and ERK 1/2 that may have complementary role in reversal of HIV-1 latency.