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Background: Tuberculosis (TB) presents serious health related complications caused by the Mycobacterium tuberculosis pathogen. Interleukin 12 (IL-12), plays a central role in T helper 1 (Th1) cells development that are implicated in chronic inflammatory pathogenesis as well as level of 1,25-dihydroxyvitamin D3 can impact on IL-12 mRNA expression at the transcriptional level. Methods: The present study included clinically confirmed 100 Mycobacterium tuberculosis cases (TB) for assessment of IL-12 mRNA expression and vitamin-D level as well as equal number of healthy controls were also included. Results: In TB cases, overall 13.01-fold higher IL-12 mRNA expression and 30.69 ng/ml vitamin-D level were observed. It was observed that higher expression of IL-12 mRNA expression was linked with TB cases had fever (p

The present work is an attempt to carry on analytical work on a biodegradable cellulosic material obtained from the fibre of a plant which is scientifically known as Grewia Optiva. In this study, 125 µm particle size of Grewia optiva fibres was used as a filtration loss controlling agent in drilling fluid. The Grewia Optiva mud samples were hot rolled at 100 °C for 16 h and it was observed that the said fibre at 5% concentration (w/v) has reduced the low pressure–low temperature (LPLT) filtration loss of drilling mud by 72.5% under the differential pressure of 100 psi. On further assessment vis a vis comparative analysis with available market products like carboxy methyl cellulose-low viscosity grade (CMC-LVG) and polyanionic cellulose-low viscosity grade (PAC-LVG), it was found that addition of CMC and PAC-dLVG at 1.4% (w/v) in reference mud, the LPLT filtration loss was reduced by 70% and 70.6%, respectively. Use of 5% GOFP in the reference mud, the high pressure-high temperature (HPHT) filtration loss of drilling fluids was reduced by 69.2% at 100 °C temperature and 500 psi pressure. The thickness of filter cake was reduced by 41.1% after the addition of 5% GOFP in the reference mud. While the conventional fluid loss reducing agents such as CMC and PAC, both at 1.4% concentration have reduced the HPHT filtration loss by 63.46% and 64.6%, respectively, as compared to reference mud. This is indicative of a valid filtration loss control potential of Grewia optiva fibre powder.

The human exposure to diverse endocrine-disrupting chemicals (EDCs) has increased dramatically over several decades with very adverse health effects. Plasticizers and flame retardants constitute important classes of EDCs interfering in endocrine physiology including the thyroid function. Thyroxine (T4) is an important hormone regulating metabolism and playing key roles in developmental processes. In this study, six phthalate and nonphthalate plasticizers and one flame retardant (BDE-153) were subjected to structural binding against thyroxine-binding globulin (TBG). The aim was to understand their potential role in thyroid dysfunction using structural binding approach. The structural study was performed using Schrodinger's induced fit docking, followed by binding energy estimations of ligands and the molecular interaction analysis between the ligands and the amino acid residues in the TBG ligand-binding pocket. The results indicated that all the compounds packed tightly into the TBG ligand-binding pocket with similar binding pattern to that of TBG native ligand, T4. A high majority of TBG interacting amino acid residues for ligands showed commonality with native ligand, T4. The estimated binding energy values were highest for BDE-153 followed by nonphthalate plasticizer, DINCH, with values comparable with native ligand, T4. The estimated binding energy values of other plasticizers DEHP, DEHT, DEHA, ATBC, and TOTM were less than DINCH. In conclusion, the tight docking conformations, amino acid interactions, and binding energy values of the most of the indicated ligands were comparable with TBG native ligand, T4, suggesting their potential for thyroid dysfunction. The results revealed highest potential thyroid disruptive action for BDE-153 and DINCH.

Autophagy is an essential cellular process that involves the transport of cytoplasmic content in double-membraned vesicles to lysosomes for degradation. Neurons do not undergo cytokinesis, and thus, the cell division process cannot reduce levels of unnecessary proteins. The primary cause of neurodegenerative disorders (NDs) is the abnormal deposition of proteins inside neuronal cells, and this could be averted by autophagic degradation. Thus, autophagy is an important consideration when considering means of developing treatments for NDs. Various pharmacological studies have reported that the active components in herbal medicines exhibit therapeutic benefits in NDs, for example, by inhibiting cholinesterase activity and modulating amyloid beta levels, and α-synuclein metabolism. A variety of bioactive constituents from medicinal plants are viewed as promising autophagy controllers and are revealed to recover the NDs by targeting the autophagic pathway. In the present review, we discuss the role of autophagy in the therapeutic management of several NDs. The molecular process responsible for autophagy and its importance in various NDs and the beneficial effects of medicinal plants in NDs by targeting autophagy are also discussed.

The present study aimed to investigate the inhibitory role of second mitochondria determined activator of caspases mimetic on inhibitor of apoptosis proteins (IAPs) and regulation of caspases in nonsmall cell lung cancer cell line.Dimethyl sulfoxide and 3-(4, 5-dimethyl thizol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay was done to determine the ICTwo doses of BV6 dependently downregulated the expression of mRNA of XIAP (P = 0.002, P= 0.0003 vs. untreated), cIAP-1 (P = 0.05, P = 0.005 vs. untreated), and cIAP-2 (P = 0.001, P = 0.0002 vs. untreated), respectively, while the compound upregulated the mRNA expression of caspase-6 (P = 0.001, P0.0001 vs. untreated) and caspase-7 (P = 0.001, P = 0.0004 vs. untreated), respectively. Dose dependent of BV6 treatment significantly decreased the protein level of XIAP (P = 0.003, P = 0.007 vs. untreated), cIAP-1 (P = 0.02, P = 0.01 vs. untreated), and cIAP-2 (P = 0.008,P = 0.008 vs. untreated), respectively. However, the compound increased the protein level of caspase-6 and caspase-7 when compared to untreated control (P = 0.006,P = 0.001) and (P = 0.01, P = 0.001), respectively.The result showed that BV6 treatment reduced the level of mRNA of XIAP, cIAP-1, and cIAP-2 and increased the gene expression of caspase-6 and caspase-7 in NCI-H23 cell line. Therefore, the study revealed that BV6 could be used in future as additional therapeutics in lung cancer.

Proteomic information revealed approximately 3,923 proteins in Mycobacterium tuberculosis H37 Rv genome of which around ∼25% of proteins are hypothetical proteins (HPs). The present work comprises computational approaches to identify and characterize the HPs of M. tuberculosis that symbolize the putative target for rationale development of a drug or antituberculosis strategy. Proteins were primarily classified based on motif and domain information, which were further analyzed for the presence of virulence factors (VFs), determination of localization, and signal peptide/enzymatic cleavage sites. 863 HPs were found, and 599 HPs were finalized based on motifs, that is, GTP (525), Trx (47), SAM (14), PE-PGRS (5), and CBD (8). 80 HPs contain virulence factor (VF), 24 HPs localized in membrane region, and 4 HPs contain signal peptide/enzymatic cleavage sites. The overall parametric study finalizes four HPs Rv0679c, Rv0906, Rv3627c, and Rv3811 that also comprise GTPase domain. Structure prediction, structure-based function prediction, molecular docking and mutation analysis of selected proteins were done. Docking studies revealed that GTP and GTPase inhibitor (mac0182344) were docked with all four proteins with high affinities. In silico point mutation studies showed that substitution of aspartate with glycine within a GTPase motif showed the largest decrease in stability and pH differentiation also affects protein's stability. This analysis thus fixes a roadmap in the direction of finding potential target of this bacterium for drug development and enlightens the efficacy of GTP as a major regulator of Mycobacterial cellular pathways.

Background Pneumocystis pneumonia (PCP) is a fatal infectious disease caused by Pneumocystis jirovecii (PJP). The major factor relevant to morbidity and mortality seems to be the host inflammatory reaction. The objective of this study was to evaluate the role of IL-2, IL-4, IL-10, and IL-13 cytokine mRNA expression among suspected P. jirovecii infection. Methods This was a cross-sectional analytical study undertaken in Aseer region, Saudi Arabia. One hundred suspected PCP cases and 100 healthy controls were included in the study. Basic clinical manifestations, radiological findings, microbiological and immunological findings were extracted from the hospital records from January 2019 to August 2019, Pneumocystis detection was done by immune-fluorescent staining (IFAT, Gomorimethanamine silver staining (GMSS), Giemsa staining, Toluidine blue O (TBO), and Pneumocystis RT-PCR. Results Increased more than 5 fold, 3 fold, 4 fold, and 7 fold of IL-2, IL-4, IL-10, and IL-13 mRNA expression were observed in PCP cases compared to controls. Higher expression of IL-2 mRNA was connected with crept, wheezing and chest X-ray findings like central perihilar infiltrate, patchy infiltrate, consolidation, hilar lymphadenopathy, pneumothorax, pleural effusion which showed higher expression compared to counterpart (pp=0.002), dyspnea (p=0.003), crept (p=0.01), and chest X-ray findings (p Conclusion Confirmed cases of P. jirovecii showed higher IL-2, IL-4, IL-10, and IL-13 mRNA expression comparatively to negative cases. Increased expression of cytokines may be indicative of infection severity and could help in patients’ management.

Background The human immunodeficiency virus (HIV)-1 latent reservoir (LR) in resting CD4+ T cells is a barrier to cure. LR measurements are commonly performed on blood samples and therefore may miss latently infected cells residing in tissues, including lymph nodes. Methods We determined the frequency of intact HIV-1 proviruses and proviral inducibility in matched peripheral blood (PB) and lymph node (LN) samples from 10 HIV-1-infected patients on antiretroviral therapy (ART) using the intact proviral DNA assay and a novel quantitative viral induction assay. Prominent viral sequences from induced viral RNA were characterized using a next-generation sequencing assay. Results The frequencies of CD4+ T cells with intact proviruses were not significantly different in PB versus LN (61/106 vs 104/106 CD4+ cells), and they were substantially lower than frequencies of CD4+ T cells with defective proviruses. The frequencies of CD4+ T cells induced to produce high levels of viral RNA were not significantly different in PB versus LN (4.3/106 vs 7.9/106), but they were 14-fold lower than the frequencies of cells with intact proviruses. Sequencing of HIV-1 RNA from induced proviruses revealed comparable sequences in paired PB and LN samples. Conclusions These results further support the use of PB as an appropriate proxy for the HIV-1 LR in secondary lymphoid organs.

In untreated HIV-1 infection, rapid viral evolution allows escape from immune responses. Viral replication can be blocked by antiretroviral therapy. However, HIV-1 persists in a latent reservoir in resting CD4(+) T cells, and rebound viremia occurs following treatment interruption. The reservoir, which is maintained in part by clonal expansion, can be measured using quantitative viral outgrowth assays (QVOAs) in which latency is reversed with T cell activation to allow viral outgrowth. Recent studies have shown that viruses detected in QVOAs prior to treatment interruption often differ from rebound viruses. We hypothesized that autologous neutralizing antibodies directed at the HIV-1 envelope (Env) protein might block outgrowth of some reservoir viruses. We modified the QVOA to reflect pressure from low concentrations of autologous antibodies and showed that outgrowth of a substantial but variable fraction of reservoir viruses is blocked by autologous contemporaneous immunoglobulin G (IgG). A reduction in outgrowth of >80% was seen in 6 of 15 individuals. This effect was due to direct neutralization. We established a phylogenetic relationship between rebound viruses and viruses growing out in vitro in the presence of autologous antibodies. Some large infected cell clones detected by QVOA carried neutralization-sensitive viruses, providing a cogent explanation for differences between rebound virus and viruses detected in standard QVOAs. Measurement of the frequency of reservoir viruses capable of outgrowth in the presence of autologous IgG might allow more accurate prediction of time to viral rebound. Ultimately, therapeutic immunization targeting the subset of variants resistant to autologous IgG might contribute to a functional cure.

HIV-1 latency is a major barrier to cure. Identification of small molecules that destabilize latency and allow immune clearance of infected cells could lead to treatment-free remission. In vitro models of HIV-1 latency involving cell lines or primary cells have been developed for characterization of HIV-1 latency and high-throughput screening for latency-reversing agents (LRAs). We have shown that the majority of LRAs identified to date are relatively ineffective in cells from infected individuals despite activity in model systems. We show here that, for diverse LRAs, latency reversal observed in model systems involves a heat shock factor 1 (HSF1)-mediated stress pathway. Small-molecule inhibition of HSF1 attenuated HIV-1 latency reversal by histone deactylase inhibitors, protein kinase C agonists, and proteasome inhibitors without interfering with the known mechanism of action of these LRAs. However, latency reversal by second mitochondria-derived activator of caspase (SMAC) mimetics was not affected by inhibition of HSF1. In cells from infected individuals, inhibition of HSF1 attenuated latency reversal by phorbol ester+ionomycin but not by anti-CD3+anti-CD28. HSF1 promotes elongation of HIV-1 RNA by recruiting P-TEFb to the HIV-1 long terminal repeat (LTR), and we show that inhibition of HSF1 attenuates the formation of elongated HIV-1 transcripts. We demonstrate that in vitro models of latency have higher levels of the P-TEFb subunit cyclin T1 than primary cells, which may explain why many LRAs are functional in model systems but relatively ineffective in primary cells. Together, these studies provide insights into why particular LRA combinations are effective in reversing latency in cells from infected individuals.

In the present investigation, the characterization of Cassia fistula leaf extracts (CFLE) mediated gold nanoparticles (CF-GNPs) and its binding features with human serum albumin (HSA) through inter...

The current study entails the design, development, and optimization of nasal mucoadhesive microspheres of lercanidipine with improved systemic bioavailability for the management of hypertension. Mucoadhesive microspheres of lercanidipine were prepared by solvent evaporation and polymerization method using bovine serum albumin (BSA) as the carrier and glutaraldehyde as the chelating agent. For screening the material and process variables, Taguchi design revealed the major influence of BSA concentration and glutaraldehyde volume as the critical factors for the preparation of microspheres. These factors were systematically optimized using Central Composite design to evaluate the particle size, entrapment efficiency, and in vitro drug release from the microspheres as the response variables. The optimized microspheres were prepared using 4.4% BSA and 0.25-mL glutaraldehyde, and stirring speed at 1500 rpm, which exhibited particle size of 34 μm, entrapment efficiency of 88.6%, Q6h of 94.67%, T60% of 3.2 h, and bioadhesion efficiency of 93.2%. In vivo pharmacokinetics in rabbits showed remarkable superiority of the optimized nasal microspheres (p

Tetrabromobisphenol A (TBBPA) is a widely used brominated flame retardant owing to its efficient fire-breaking property. However, leaching of TBBPA into the environment has been a global health concern due to the endocrine-disrupting activity (EDA) associated with TBBPA exposure. Limited studies are available on the hazardous effects of TBBPA on reproductive function. The aim of the present study was the structural characterization of potential EDA of TBBPA in reproductive hormone signaling and transport including steroid nuclear receptors, such as estrogen receptor alpha (ERα), estrogen receptor beta (ERβ), androgen receptor (AR), progesterone receptor (PR), and the steroid transport protein, sex hormone-binding globulin (SHBG). The structural binding characterization of TBBPA with the sex steroid nuclear receptors and transport protein was performed by induced-fit docking using the Schrödinger 2017 suite. The results revealed that the TBBPA binding pattern and molecular interactions with the indicated receptors and transport protein displayed overall similarity with their respective native ligands. The estimated binding energy value of TBBPA for ERα was similar to the native ligand, estradiol, indicating tight binding and greater potential for TBBPA to disrupt ERα signaling. For ERβ, AR, PR and SHBG, the estimated binding energy values were also close to their respective native ligands, indicating potential for interference in native hormone signaling and transport. In conclusion, TBBPA exposure in humans may potentially cause disruption of sex steroid signaling and transport, and thus lead to reproductive dysfunction.

Cancers of the lymphatic system are broadly classified into Hodgkin and non‐Hodgkin types. Although lymphomas can be effectively treated with chemotherapy, this approach is associated with the risk of adverse side effects. High intake of certain vegetables and fruits is associated with a reduced risk of cancer development. We hypothesized that Annona fruit, which is rich in fibers and phytochemicals that are known to possess anticancer properties, can be effective in inhibiting lymphoma growth. The Annona fruit's fractions were extracted with water, methanol, or chloroform and then assayed for total phenolic, flavonoids, and tannins content; antioxidation activities; and inhibition of in vitro cell proliferation using the Ramos‐1 lymphoma cells. The methanol fractions contained the highest phenolics, flavonoids, and tannins content, and antioxidation activity. However, the methanol extracts of skin, pulp, and seeds had a moderate whereas the chloroform extracts of pulp and seeds had strong effects on Ramos‐1 cell proliferation. Our findings suggest that Annona fruits may be effective in the prevention or treatment of lymphoma., The methanol fractions of Annona fruits contained the highest phenolic content and antioxidation activity. However, the methanol extracts of skin, pulp, and seeds had a moderate whereas the chloroform extracts of pulp and seeds had strong effects on Ramos‐1 cell proliferation. Our findings suggest that Annona fruits may be effective in the prevention or treatment of lymphoma.

Gut microbiota of humans harbour a complex and dynamic community of more than 100 trillion beneficial microbes. Recent advances in tools and techniques of microbial analysis have made it possible to understand better the important role played by gut microbiota in the human body. The composition of gut microbiota depends on many factors, such as the mode of delivery of the baby, environmental conditions, antibiotic usage, age, and diet. Homeostasis of gut microbiota is important for maintaining good health as it influences human nutrition, metabolism, and immune function, while dysbiosis of the gut microbiota is associated with various gastrointestinal and metabolic disorders, such as inflammatory bowel disease, obesity, type 2 diabetes, colon cancer, and dysregulation of the immune system. Clinical evidences have shown that diseases caused by imbalance gut microbiota can be treated by probiotics. However, more research work is required to get insightful knowledge of what kind of particular microbes and underlying molecular mechanisms are responsible for a particular disease.

The aim of this study was to investigate the COVID-19 vaccination acceptance rate and its determinants among healthcare workers in a multicenter study. This was a cross-sectional multi-center survey conducted from February 5 to April 29, 2021. The questionnaire consisted of 26 items in 6 subscales. The English version of the questionnaire was translated into seven languages and distributed through Google Forms using snowball sampling; a colleague in each country was responsible for the forward and backward translation, and also the distribution of the questionnaire. A forward stepwise logistic regression was utilized to explore the variables and questionnaire factors tied to the intention to COVID-19 vaccination. 4630 participants from 91 countries completed the questionnaire. According to the United Nations Development Program 2020, 43.6 % of participants were from low Human Development Index (HDI) regions, 48.3 % high and very high, and 8.1 % from medium. The overall vaccination hesitancy rate was 37 %. Three out of six factors of the questionnaire were significantly related to intention to the vaccination. While ���Perceived benefits of the COVID-19 vaccination��� (OR: 3.82, p-value, EXCLI Journal; 21:Doc93; ISSN 1611-2156

The molecular characteristics of metastatic upper tract urothelial carcinoma (UTUC) are unknown. The genomic and transcriptomic differences between primary and metastatic UTUC is not well described either. We combined whole-exome sequencing, RNA-sequencing, and Imaging Mass Cytometry™ (IMC™) of 44 tumor samples from 28 patients with high-grade primary and metastatic UTUC. IMC enables spatially resolved single-cell analyses to examine the evolution of cancer cell, immune cell, and stromal cell markers using mass cytometry with lanthanide metal-conjugated antibodies. We discovered that actionable genomic alterations are frequently discordant between primary and metastatic UTUC tumors in the same patient. In contrast, molecular subtype membership and immune depletion signature were stable across primary and matched metastatic UTUC. Molecular and immune subtypes were consistent between bulk RNA-sequencing and mass cytometry of protein markers from 340,798 single-cells. Molecular subtyping at the single cell level was highly conserved between primary and metastatic UTUC tumors within the same patient.

Tocopherols are lipid-soluble compounds regarded as vitamin E compounds and they function as antioxidants in scavenging lipid peroxyl radicals and quenching reactive oxygen species (ROS). In our previous studies, we isolated five tocopherol biosynthesis genes from sweetpotato (Ipomoea batatas [L.] Lam) plants including 4-hydroxyphenylpyruvate dioxygenase (IbHPPD). HPPD is the first regulatory enzyme in vitamin E biosynthesis and serves to catalyze in the first steps α-tocopherol and plastoquinone biosynthesis by converting 4-hydroxyphenylpyruvate (HPP) to homogentisic acid (HGA). In this study, we generated transgenic sweetpotato plants overexpressing IbHPPD under the control of cauliflower mosaic virus (CaMV) 35S promoter (referred to as HP plants) via Agrobacterium-mediated transformation to understand the function of IbHPPD in sweetpotato. Three transgenic lines (HP3, HP14 and HP15) with high transcript levels of IbHPPD were selected for further characterization. Compared with non-transgenic (NT) plants, HP plants exhibited enhanced tolerance to multiple environmental stresses, including salt, drought, and oxidative stresses. In addition, HP plants showed increased tolerance to the herbicide sulcotrione, which is involved in the inhibition of the HPPD. Interestingly, after stress treatments, HP plants also showed higher abscisic acid (ABA) contents than NT plants. Under dehydrated condition, HP plants displayed an elevated α-tocopherol content to 19-27% in leaves compared with NT plants. These results indicate that increased abiotic stress tolerance in HP plants is related to inducing enhancement of α-tocopherol and ABA contents.

Background Malaria is a life-threatening, multisystem disease caused by the plasmodial parasite with a global incidence of approximately 229 million annually. The parasites are known to have unique and crucial interactions with various body tissues during its life cycle, notably the liver, spleen, and recent work has shown the bone marrow to be a reservoir of infection. Methods This study is a case series of patients in whom examination of bone marrow revealed malarial parasites. A retrospective record review of 35 parasite-positive bone marrow specimens examined at Aga Khan University Hospital (AKUH), Karachi, Pakistan, over the years 2007 to 2015 was conducted. Bone marrow aspirates were collected as per International Council for Standardization in Haematology (ICSH) guidelines. Results The median age of patients was 22 years (range 1–75), and 60 % (n = 21) were male. 22 patients had evidence of Plasmodium falciparum, 12 had evidence of Plasmodium vivax and 1 patient had a mixed infection. Gametocytes and trophozoites were the most common stages identified on both peripheral blood and bone marrow examinations. Indications for bone marrow examination included fever of unknown origin and the workup of cytopenias and malignancies. Conclusions The incidental finding of Plasmodium in samples of bone marrow suggests the reticuloendothelial system may be regularly harbour these parasites, be the infection acute or chronic in character.

Background Musculoskeletal tuberculosis forms 10–25% of extrapulmonary tuberculosis which mainly involves the spine or weight-bearing joints. Tuberculous involvement of the sternum is a rare clinical entity even in countries where tuberculosis has high prevalence. Primary tuberculous sternal osteomyelitis accounts for approximately 0.3% of all types of tubercular osteomyelitis and the probable source appears to be extension from paratracheal or hilar lymph nodes. Despite tuberculosis being a common disease in endemic countries and worldwide, a thorough literature search of the PubMed database for keywords “primary tuberculosis of sternum” and “primary tuberculous osteomyelitis of sternum” yielded 30 and 22 articles, respectively. Case presentation We present an unusual case of a large dumb-bell-shaped cold abscess arising due to infection of the sternum. A 23-year-old immunocompetent Asian woman presented with a gradually progressing painless swelling on anterior chest wall for the last 5 months. She had a large visible swelling on anterior chest wall which was 12.5 cm in diameter, soft, non-tender, temperature was not raised, and fluctuant. Magnetic resonance imaging showed a large dumb-bell-shaped hyperintense collection in upper anterior chest wall with marrow edema and cortical irregularity in left side of manubrium. Pus was positive for nucleic acid testing (cartridge-based nucleic acid amplification test) for Mycobacterium tuberculosis and later culture was also positive. She was started on anti-tubercular therapy and aspirated twice. Currently, she has completed 6 months of therapy and the swelling has now disappeared. Discussion Swelling, pain localized to sternum, or ulceration of the skin with discharging sinus along with or without constitutional symptoms are the usual presentation. A high element of suspicion is needed for early diagnosis and treatment to prevent its complications. Sternal mycobacterial infections are categorized as primary, secondary, and/or acquired postoperatively. Although radiological investigations aid in diagnosis, the diagnosis is established by positive culture or histopathological examination. Anti-tubercular therapy is the mainstay of treatment with standard four-drug regimen for 6–9 months. Surgical drainage of the abscess should be considered only if it does not resolve by aspiration and anti-tubercular therapy.

The Anaphase Promoting Complex (APC) coactivator Cdh1 drives proper cell cycle progression and is implicated in the suppression of tumorigenesis. However, it remains elusive how Cdh1 restrains cancer progression and how tumor cells escape the inhibition of Cdh1. Here we report that Cdh1 suppresses the kinase activity of c-Src in an APC-independent manner. Depleting Cdh1 accelerates breast cancer cell proliferation and cooperates with PTEN loss to promote breast tumor progression in mice. Hyperactive c-Src, on the other hand, reciprocally inhibits the ubiquitin E3 ligase activity of APCCdh1 through direct phosphorylation of Cdh1 at its N-terminus, which disrupts the interaction between Cdh1 and the APC core complex. Furthermore, pharmacological inhibition of c-Src restores APCCdh1 tumor suppressor function to repress a panel of APCCdh1 oncogenic substrates. Our findings reveal a reciprocal feedback circuit of Cdh1 and c-Src in the crosstalk between the cell cycle machinery and the c-Src signaling pathway., The Anaphase Promoting Complex adaptor protein Cdh1 tightly controls cell cycle progression to restrain tumorigenesis but the mechanism is not completely known. Here, the authors show that reciprocal inhibition between Cdh1 and the c-Src signaling pathway regulate breast cancer tumorigenesis.

The antifungal properties shown by different higher plant extracts are due to the presence of phytochemicals. The activities of Alangium solviifolium (L.f.) Wangerin root in controlling two fungal species that belong to Alternaria spp- Alternaria brassicae and A. brassicicola, the causal organisms of leaf blight of crucifers was evaluated in vitro assay. The aqueous extracts of different parts of the tested plant such as bark, stem, root, leaf and flower were screened against test fungi. Among them, the results of the study revealed that the aqueous extracts of A. solviifolium(L.f.) Wangerin root showed the inhibition of mycelial growth against test fungi completely (100%). The phytochemical analysis of active plant has revealed that the presence of some chemical compounds as alkaloids, terpenoids, phenol/tannins, saponins, flavonoids and glycosides in both aqueous and ethanol extracts is likely to be responsible for the antifungal activity against test fungi. This study confirms the presence of various bioactive compounds of an active plant.

We have reported the eco-friendly gold nanoparticles (AuNPs) from 15% aqueous leaf extract of therapeutic plant Nymphaea nouchali where leaf extract acts as both reducing and stabilizing agents. The generation of AuNPs was confirmed by UV–visible spectroscopy, FTIR, FE-SEM, DLS, and the size and morphology were further confirmed via TEM analysis (average d = 54.7 nm). The existence of gold signal in EDX spectrum and measurement of zeta potential reveals a negative value (−21.7 mV) clearly indicates its stability. This synthesized AuNPs shows strong in vitro bactericidal effect against E. coli (DH5-Alpha) with 0.12 nM minimum inhibitory concentration, suggests its progress in therapeutic efficacy. The interaction of this AuNPs with human serum albumin (HSA) was investigated by UV–visible, FTIR, fluorescence, circular dichroism spectroscopies and zeta potential evaluation at 25 °C. It has been observed that the interaction mechanism is a static quenching process with the slightest change in the HSA secondary conformation because of HSA-AuNPs conjugate formation, but the primary structure remains unaffected. Negative free energy change indicates, the HSA-AuNPs formation is a thermodynamically favourable process. This analysis may offer a valuable information on the healthier antibacterial AuNPs synthesized from a medicinal plant extract.

For several decades antibiotics are used to combat against pathogenic bacteria, but their misuse and overuse have caused the emergence of resistant bacteria. The scarcities of effective antibiotics along with unavailability of alternative solutions have exacerbated bacterial infections and mortality rate. This review provides the concept of bacterial resistome and mechanisms of resistance. It has also described the utility of whole genome sequencing in identifying resistance and its dissemination in association with available bioinformatics tools and databases. Moreover, the whole genome sequencing methodology described in this review will help to select effective antibiotics, maintain unparalleled surveillance of resistance and provide early diagnosis during resistance outbreaks. The provided information could be used to control infection caused by resistant microorganisms.

A stable latent reservoir for HIV-1 in resting CD4+ T cells is the principal barrier to a cure1-3. Curative strategies that target the reservoir are being tested4,5 and require accurate, scalable reservoir assays. The reservoir was defined with quantitative viral outgrowth assays for cells that release infectious virus after one round of T cell activation1. However, these quantitative outgrowth assays and newer assays for cells that produce viral RNA after activation6 may underestimate the reservoir size because one round of activation does not induce all proviruses7. Many studies rely on simple assays based on polymerase chain reaction to detect proviral DNA regardless of transcriptional status, but the clinical relevance of these assays is unclear, as the vast majority of proviruses are defective7-9. Here we describe a more accurate method of measuring the HIV-1 reservoir that separately quantifies intact and defective proviruses. We show that the dynamics of cells that carry intact and defective proviruses are different in vitro and in vivo. These findings have implications for targeting the intact proviruses that are a barrier to curing HIV infection.

Alzheimer's disease (AD) is a chronic neurodegenerative disease that causes memory loss and decline in cognitive abilities. AD is the sixth leading cause of death in the United States, affecting an estimated 5 million Americans. To assess the association between multiple genetic variants and multiple measurements of structural changes in the brain a recent study of AD used a multivariate measure of linear dependence, the RV coefficient. The authors decomposed the RV coefficient into contributions from individual variants and displayed these contributions graphically. We investigate the properties of such a `contribution plot' in terms of an underlying linear model, and discuss estimation of the components of the plot when the correlation signal may be sparse. The contribution plot is applied to simulated data and to genomic and brain imaging data from the Alzheimer's Disease Neuroimaging Initiative., Comment: 20 pages, 8 figures

Background: The epidermal growth factor receptor 1 (EGFR1) plays a significant role in cell proliferation and development. Its regulation in humans is very critical and incompletely understood in Non small cell lung cancer (NSCLC). Methods: 100 newly diagnosed NSCLC (lung adenocarcinoma) patients and 100 healthy controls were included and allele specific (AS) polymerase chain reaction (PCR) was used to genotype and expression was analyzed by quantitative real time PCR. Overall survival of patients was analyzed by Kaplan-Meier method and for prognostic significance ROC curve was plotted. Results: A statistically significant difference (p

Significance: Head and neck squamous cell cancer (HNSCC) is a complex disease characterized by high genetic and metabolic heterogeneity. Radiation therapy (RT) alone or combined with systemic chemotherapy is widely used for treatment of HNSCC as definitive treatment or as adjuvant treatment after surgery. Antibodies against epidermal growth factor receptor are used in definitive or palliative treatment. Recent Advances: Emerging targeted therapies against other proteins of interest as well as programmed cell death protein 1 and programmed death-ligand 1 immunotherapies are being explored in clinical trials. Critical Issues: The disease heterogeneity, invasiveness, and resistance to standard of care RT or chemoradiation therapy continue to constitute significant roadblocks for treatment and patients' quality of life (QOL) despite improvements in treatment modality and the emergence of new therapies over the past two decades. Future Directions: As reviewed here, alterations in redox metabolism occur at all stages of HNSCC management, providing opportunities for improved prevention, early detection, response to therapies, and QOL. Bioinformatics and computational systems biology approaches are key to integrate redox effects with multiomics data from cells and clinical specimens and to identify redox modifiers or modifiable target proteins to achieve improved clinical outcomes. Antioxid. Redox Signal.

Herein, we have shown the effect of cations and anions of three different ionic liquids (ILs); 1-Decyl-3-methylimidazolium chloride [DMIM][Cl], 1-Butyl-1-methylpyrrolidinium tetrafluoroborate [BMP][BF4] and 1-Butyl-1-methylpyrrolidinium bromide [BMP][Br] on the stability and activity of hen egg white lysozyme (HEWL). The spectroscopic techniques; fluorescence, time resolved fluorescence, UV–visible, CD spectroscopy and molecular docking method were applied to investigate the differential behavior of HEWL in the aqueous solution of these ILs. Fluorescence results showed that imidazolium-based IL binds more strongly than pyrrolidinium based ILs with HEWL. The docking results suggested that these ILs bind at the active site of HEWL mainly through electrostatic, hydrophobic and van der Waals interactions. Results revealed that depending upon cations and anions, these ILs exert concentration dependent effect on HEWL. The conformational stability of HEWL is maintained in [BMP][BF4] and [BMP][Br] ILs while it decreases with increasing concentration of [DMIM][Cl]. A slight increase of 1 °C in melting temperature (Tm) of HEWL is obtained in presence of [BMP][BF4] and [BMP][Br] ILs while a much decrease of about 16.9 °C in Tm is obtained in presence of [DMIM][Cl]. Surprisingly, different effects of these ILs were observed on the activity of HEWL against Gram-positive bacteria; Micrococcus lysodeikticus. The activity of HEWL increases with increasing concentration of [BMP][BF4] and [DMIM][Cl] up to certain concentration range while it decreases with increasing concentration of [BMP][Br]. Comparatively, higher activity is obtained in presence of [DMIM][Cl] than [BMP][BF4].

Barcoding of Cycas pschannae Srivast. & Singh from Andaman & Nicobar Islands was done with the help of rbcL and ITS(Standard Universal Barcode Sequence- CBoL) and submitted to GenBank. Phylogenetic relationship of this species was analyzed and Cladistic study of nine Cycas species from India and adjacent country including Cycas revoluta thunb had been conducted with help of BLAST tool by using rbcL barcode sequence, which reveals that Cycas pschannae has close relation with Cycas circinalis as compared to other Indian species.

Background K13 propeller (k13) polymorphism are useful molecular markers for tracking the emergence and spread of artemisinin resistance in Plasmodium falciparum. Polymorphisms are reported from Cambodia with rapid invasion of the population and almost near fixation in south East Asia. The study describes single nucleotide polymorphisms in Kelch protein propeller domain of P. falciparum associated with artemisinin resistance from Southern Pakistan. Methods Two hundred and forty-nine samples were collected from patients with microscopy confirmed P. falciparum malaria attending Aga Khan University Hospital during September 2015-April 2018. DNA was isolated using the whole blood protocol for the QIAmp DNA Blood Kit. The k13 propeller gene (k13) was amplified using nested PCR. Double-strand sequencing of PCR products was performed using Sanger sequencing methodology. Sequences were analysed with MEGA 6 and Bio edit software to identify specific SNP combinations. Results All isolates analysed for k13 propeller allele were observed as wild-type in samples collected post implementation of ACT in Pakistan. C580Y, A675V, Y493H and R539T variants associated with reduced susceptibility to artemisinin-based combination therapy (ACT) were not found. Low frequency of M476I and C469Y polymorphisms was found, which is significantly associated with artemisinin resistance. Conclusion Low frequencies of both nonsynonymous and synonymous polymorphisms were observed in P. falciparum isolates circulating in Southern Pakistan. The absence of known molecular markers of artemisinin resistance in this region is favourable for anti-malarial efficacy of ACT. Surveillance of anti-malarial drug resistance to detect its emergence and spread need to be strengthened in Pakistan.

Epidemiological studies have established that untreated hypertension (HTN) is a major independent risk factor for developing cardiovascular diseases (CVD), stroke, renal failure, and other conditions. Several important studies have been published to prevent and manage HTN; however, antihypertensive agents' optimal choice remains controversial. Therefore, the present study is undertaken to update our knowledge in the primary treatment of HTN, specifically in the setting of other three important diseases. MicroRNAs (miRNAs) are remarkably stable short endogenous conserved non-coding RNAs that bind to the mRNA at its (3′ UTR) to regulate its gene expression by causing translational repression or mRNA degradation. Through their coordinated activities on different pathways and networks, individual miRNAs control normal and pathological cellular processes. Therefore, to identify the critical miRNA-mRNA-TF interactions, we performed systematic bioinformatics analysis. We have also employed the molecular modelling and docking approach to identify the therapeutic target that delivers novel empathies into Food and Drug Administration approved and herbal drug response physiology. Gene Expression Omnibus (GEO) was employed to identify the differentially expressed genes (DEGs) and hub genes- KNG1, HLA-DPB1, CXCL8, IL1B, and BCL2. The HTN associated feed-forward loop (FFL) network included miR-9-5p, KNG1 and AR. We employed high throughput screening to get the best interacting compounds, telmisartan and limonin, that provided a significant docking score (−13.3 and −12.0 kcal/mol) and a potential protective effect that may help to combat the impact of HTN. The present study provides novel insight into HTN etiology through the identification of mRNAs and miRNAs and associated pathways.

Background: There is strong evidence that disease progression, drug response and overall clinical outcome of CML disease are not only decided by BCR/ABL1 oncoprotein but depend on accumulation of additional genetic and epigenetic aberrations. DNA hydroxymethylation is implicated in the development of variety of diseases. DNA hydroxymethylation in gene promoters plays an important role in disease progression, drug response and clinical outcome of various diseases. Therefore in this study, we aimed to explore the role of aberrant hydroxymethylation in promoter regions of different tumor suppressor genes in relation to CML disease progression, response to imatinib therapy and clinical outcome. We recruited 150 CML patients at different clinical stages of the disease. Patients were followed up for 48 months and haematological/molecular responses were analysed. Haematological response was analysed by peripheral blood smear. BCR/ABL1 specific TaqMan probe based qRT-PCR was used for assessing the molecular response of CML patients on imatinib therapy. Promoter hydroxymethylation of the genes was characterized using MS-PCR.Results: We observed that promoter hydroxymethylation of cell cycle regulating and apoptosis related genes characterize advanced CML disease and poor imatinib respondents. Although, cytokine signalling (SOCS1) gene was hypermethylated in advanced stages of CML and accumulated in patients with poor imatinib response, but the differences were not statistically significant. Moreover, we found hypermethylation of cell cycle regulating genes (p14ARF, RASSF1 and p16INK4A) and cytokine signalling gene (SOCS1) significantly associated with poor overall survival of CML patients on imatinib therapy. The results of this study are in agreement of the role of aberrant DNA methylation of different tumor suppressor genes as potential biomarkers of CML disease progression, poor imatinib response and overall clinical outcome. Conclusion: In this study, we report that promoter hydroxymethylation of cell cycle and apoptosis related genes is a characteristic feature of CML disease progression, defines poor imatinib respondents and poor overall survival of CML patients to imatinib therapy.

Clonal expansion of infected CD4(+) T cells is a major mechanism of HIV-1 persistence and a barrier to achieving a cure. Potential causes are homeostatic proliferation, effects of HIV-1 integration, and interaction with antigens. Here, we show that it is possible to link antigen responsiveness, the full proviral sequence, the integration site, and the T cell receptor β-chain (TCRβ) sequence to examine the role of recurrent antigenic exposure in maintaining the HIV-1 reservoir. We isolated CMV- and Gag-responding CD4(+) T cells from 10 treated individuals. Proviral populations in CMV-responding cells were dominated by large clones, including clones harboring replication-competent proviruses. TCRβ repertoires showed high clonality driven by converging adaptive responses. Although some proviruses were in genes linked to HIV-1 persistence (BACH2, STAT5B, MKL1), the proliferation of infected cells under antigenic stimulation occurred regardless of the site of integration. Paired TCRβ and integration site analysis showed that infection could occur early or late in the course of a clone’s response to antigen and could generate infected cell populations too large to be explained solely by homeostatic proliferation. Together, these findings implicate antigen-driven clonal selection as a major factor in HIV-1 persistence, a finding that will be a difficult challenge to eradication efforts.

Borage oil that is extracted from (Borago officinalis Linn.) is a well-known medicinal plant having various medicinal benefits. In this work, an affordable, simple, reliable, rapid and easily accessible high-performance thin-layer chromatography (HPTLC) method was developed for the estimation of gamma-linolenic acid (GLA) in borage oil. HPTLC method employs thin-layer chromatography (TLC) aluminum plates precoated with silica gel (G60F254) as the stationary phase, and the mixture of hexane:toulene:glacial acetic acid (3:7:1, v/v/v) was used as the mobile phase. Densitometric analysis of the TLC plates was carried out at 200 nm. The developed method showed well-resolved spots with retention factor (Rf) value of 0.53 ± 0.04 for GLA. Various experimental conditions like saturation time for chamber, solvent phase migration and width of the band were studied intensely for selecting the optimum conditions. The method validation was performed for parameters like linearity, accuracy, specificity and precision. The values of limit of detection and limit of quantification for GLA were found to be 0.221 and 0.737 μg/band, respectively. In nutshell, the developed HPTLC method was found to be highly sensitive for the estimation of GLA in the herbal oil samples and formulations.

Antibody-drug conjugates (ADCs) are a new generation of anticancer therapeutics with enhanced selectivity against the target cancer cells over the conventional chemotherapeutic agents. Such conjugates help in delivery of drugs with enhanced therapeutic performance, thus creating a highly target-specific and cytotoxic-anticancer therapy with the advantage of a wide therapeutic window. Several drug molecules have been investigated in this regard for improving their anticancer activity with the help of antibodies of specific targeting ability to the human cancers. ADCs also help in reducing the toxicity of chemotherapeutic agents by transforming them into the inactive agents.

Highlights • NGS-based clinical studies have reported detection of clinically relevant alterations in ∼30% of patients. • To increase the detection of potential targets, we integrated whole-exome sequencing with a multiplex PCR-based NGS assay for fusion detection. • The targeted transcriptome sequencing was performed using a very low RNA input from archival cancer tissues. • With this integrated approach, we demonstrate a significant increase in detection of targetable genomic alterations in cancer patients., Background Frequency of clinically relevant mutations in solid tumors by targeted and whole-exome sequencing is ∼30%. Transcriptome analysis complements detection of actionable gene fusions in advanced cancer patients. Goal of this study was to determine the added value of anchored multiplex PCR (AMP)-based next-generation sequencing (NGS) assay to identify further potential drug targets, when coupled with whole-exome sequencing (WES). Methods Selected series of fifty-six samples from 55 patients enrolled in our precision medicine study were interrogated by WES and AMP-based NGS. RNA-seq was performed in 19 cases. Clinically relevant and actionable alterations detected by three methods were integrated and analyzed. Results AMP-based NGS detected 48 fusions in 31 samples (55.4%); 31.25% (15/48) were classified as targetable based on published literature. WES revealed 29 samples (51.8%) harbored targetable alterations. TMB-high and MSI-high status were observed in 12.7% and 1.8% of cases. RNA-seq from 19 samples identified 8 targetable fusions (42.1%), also captured by AMP-based NGS. When number of actionable fusions detected by AMP-based NGS were added to WES targetable alterations, 66.1% of samples had potential drug targets. When both WES and RNA-seq were analyzed, 57.8% of samples had targetable alterations. Conclusions This study highlights importance of an integrative genomic approach for precision oncology, including use of different NGS platforms with complementary features. Integrating RNA data (whole transcriptome or AMP-based NGS) significantly enhances detection of potential targets in cancer patients. In absence of fresh frozen tissue, AMP-based NGS is a robust method to detect actionable fusions using low-input RNA from archival tissue.

Traditionally, lipolysis has been regarded as an enzymatic activity that liberates fatty acids as metabolic fuel. However, recent work has shown that novel substrates, including a variety of lipid compounds such as fatty acids and their derivatives, release lipolysis products that act as signaling molecules and transcriptional modulators. While these studies have expanded the role of lipolysis, the mechanisms underpinning lipolysis signaling are not fully defined. Here, we uncover a new mechanism regulating glucose uptake, whereby activation of lipolysis, in response to elevated cAMP, leads to the stimulation of thioredoxin-interacting protein (TXNIP) degradation. This, in turn, selectively induces glucose transporter 1 surface localization and glucose uptake in 3T3-L1 adipocytes and increases lactate production. Interestingly, cAMP-induced glucose uptake via degradation of TXNIP is largely dependent upon adipose triglyceride lipase (ATGL) and not hormone-sensitive lipase or monoacylglycerol lipase. Pharmacological inhibition or knockdown of ATGL alone prevents cAMP-dependent TXNIP degradation and thus significantly decreases glucose uptake and lactate secretion. Conversely, overexpression of ATGL amplifies the cAMP response, yielding increased glucose uptake and lactate production. Similarly, knockdown of TXNIP elicits enhanced basal glucose uptake and lactate secretion, and increased cAMP further amplifies this phenotype. Overexpression of TXNIP reduces basal and cAMP-stimulated glucose uptake and lactate secretion. As a proof of concept, we replicated these findings in human primary adipocytes and observed TXNIP degradation and increased glucose uptake and lactate secretion upon elevated cAMP signaling. Taken together, our results suggest a crosstalk between ATGL-mediated lipolysis and glucose uptake.

Providing health information is a non-pharmaceutical intervention designed to reduce disease transmission and infection risk by encouraging behavior change. But does knowledge change behavior? We test whether coronavirus health knowledge promotes protective risk mitigation behaviors early in the Covid-19 pandemic in samples from four African countries (Ghana, Malawi, Sierra Leone, and Tanzania). Despite reputations for weak health sectors and low average levels of education, health knowledge of the symptoms and transmission mechanisms was high in all countries in the two months after the virus entered the country. Higher knowledge was associated with increased protective measures that would likely lower disease risk with one exception-knowledge was inversely correlated with social distancing. Respondents largely adhered to mask mandates and lockdowns, but continued coming into contact with others at small, informal gatherings, gatherings not affected by mandates. Knowledge alone did not reduce all risky activities, especially gatherings within other people's homes. Even early in the pandemic, income loss or stress were commonly reported. Our results suggest that early and consistent government provision of health information likely reduced the initial severity of the pandemic in Africa but was not a panacea.

Plasmodium vivax (P. vivax) is the most common cause of malaria in Pakistan. Several cases of severe malaria due to P.vivax have been reported from Pakistan and India, however morphological characteristics of the parasite have been mainly ignored. We present two cases of P. vivax mono-infection, which were characterized by multiple infected red blood cells, similar to that seen in Plasmodium falciparum, as observed under microscopy. Both cases were confirmed as mono-infection of P.vivax on Giemsa stained thick and thin films, malaria rapid diagnostic test (RDT) and Polymerase Chain Reaction (PCR). Morphology on peripheral blood smear remains the gold standard for diagnosis of malaria and mimicking morphological features leads to misdiagnosis and mismanagement of patients.