Activation and differentiation of cognate T cells by a dextran-based antigen-presenting system for cancer immunotherapy.

Immunotherapeutic modulation of antigen-specific T-cell responses instead of the whole repertoire helps avoid immune-related adverse events. We have developed an artificial antigen-presenting system (aAPS) where multiple copies of a multimeric peptide-MHC class I complex presenting a murine class I MHC restricted ovalbumin-derived peptide (signal 1), along with a costimulatory ligand (signal 2) are chemically conjugated to a dextran backbone. Cognate naive CD8+ T cells, when treated with this aAPS underwent significant expansion and showed an activated phenotype. Furthermore, elevated expression of effector cytokines led to the differentiation of these cells to cytotoxic T lymphocytes which resulted in target cell lysis, indicative of the functional efficacy of the aAPS. CD8+ T cells with decreased proliferative potential due to repeated antigenic stimulation could also be re-expanded by the developed aAPS. Thus, the developed aAPS warrants further engineering for future application as a rapidly customizable personalized immunotherapeutic agent, incorporating patient-specific MHC-restricted tumor antigens and different costimulatory signals to modulate both naive and antigen-experienced but exhausted tumor-specific T cells in cancer.

Magnetic Stress-Driven Metal-Insulator Transition in Strongly Correlated Antiferromagnetic CrN.

Traditionally, the Coulomb repulsion or Peierls instability causes the metal-insulator phase transitions in strongly correlated quantum materials. In comparison, magnetic stress is predicted to drive the metal-insulator transition in materials exhibiting strong spin-lattice coupling. However, this mechanism lacks experimental validation and an in-depth understanding. Here we demonstrate the existence of the magnetic stress-driven metal-insulator transition in an archetypal material, chromium nitride. Structural, magnetic, electronic transport characterization, and first-principles modeling analysis show that the phase transition temperature in CrN is directly proportional to the strain-controlled anisotropic magnetic stress. The compressive strain increases the magnetic stress, leading to the much-coveted room-temperature transition. In contrast, tensile strain and the inclusion of nonmagnetic cations weaken the magnetic stress and reduce the transition temperature. This discovery of a new physical origin of metal-insulator phase transition that unifies spin, charge, and lattice degrees of freedom in correlated materials marks a new paradigm and could lead to novel device functionalities.

In situ N K-edge XANES study of iron, cobalt and nickel nitride thin films.

A prototype in situ X-ray absorption near-edge structure (XANES) system was developed to explore its sensitivity for ultra-thin films of iron-nitride (Fe-N), cobalt-nitride (Co-N) and nickel-nitride (Ni-N). They were grown using DC-magnetron sputtering in the presence of an N2 plasma atmosphere at the experimental station of the soft XAS beamline BL01 (Indus-2, RRCAT, India). XANES measurements were performed at the N K-edge in all three cases. It was found that the N K-edge spectral shape and intensity are greatly affected by increasing thickness and appear to be highly sensitive, especially in low-thickness regions. From a certain thickness of ∼1000 Å, however, samples exhibit a bulk-like behavior. On the basis of the obtained results, different growth stages were identified. Furthermore, the presence of a molecular N2 component in the ultra-thin regime (<100 Å) was also obtained in all three cases studied in this work. In essence, this prototype in situ system reveals that N K-edge XANES is a powerful technique for studying ultra-thin films, and the development of a dedicated in situ system can be effective in probing several phenomena that remain hitherto unexplored in such types of transition metal nitride thin films.

Discovery of novel 1,3,5-triazine as adenosine A2A receptor antagonist for benefit in Parkinson's disease.

Parkinson's disease (PD) is a chronic neuro-degenerative ailment characterized by impairment in various motor and nonmotor functions of the body. In the past few years, adenosine A2 A receptor (A2 AR) antagonists have attracted much attention due to significant relief in PD. Therefore, in the current study, we intend to disclose the development of novel 1,3,5-triazines as A2 AR antagonist. The radioligand binding and selectivity of analogs were tested in HEK293 (human embryonic kidney) and the cells were transfected with pcDNA 3.1(+) containing full-length human A2 AR cDNA and pcDNA 3.1(+) containing full-length human A1 R cDNA, where they exhibit selective affinity for A2 AR. Molecular docking analysis was also conducted to rationalize the probable mode of action, binding affinity, and orientation of the most potent molecule (7c) at the active site of A2 AR. It has been shown that compound 7c form numerous nonbonded interactions in the active site of A2 AR by interacting with Ala59, Ala63, Ile80, Val84 Glu169, Phe168, Met270, and Ile274. The study revealed 1,3,5-triazines as a novel class of A2 AR antagonists.

Discovery of novel pyrazole derivatives as a potent anti-inflammatory agent in RAW264.7 cells via inhibition of NF-ĸB for possible benefit against SARS-CoV-2.

Due to unavailability of a specific drug/vaccine to attenuate severe acute respiratory syndrome coronavirus 2, the current strategy to combat the infection has been largely dependent upon the use of anti-inflammatory drugs to control cytokines storm responsible for respiratory depression. Thus, in this study, we discovered novel pyrazole analogs as a potent nuclear factor kappa B (NF-ĸB) inhibitor. The compounds were assessed for NF-ĸB transcriptional inhibitory activity in RAW264.7 cells after stimulation with lipopolysaccharides (LPS), revealing Compound 6c as the most potent analog among the tested series. The effect of Compound 6c was further investigated on the levels of interleukin-1ß, tumor necrosis factor-α, and interleukin-6 in LPS-stimulated RAW267.4 cells by enzyme immunoassay, where it causes a significant reduction in the level of these cytokines. In Western blot analysis, Compound 6c also causes the inhibition of inhibitor kappa B-α and NF-κB. It was found to be snugly fitted into the inner grove of the active site of NF-ĸB by forming H-bonds and a nonbonded interaction with Asn28 in a docking analysis.

Evaluation of antioxidant and antimicrobial potential of a novel Himalayan plant Reinwardtia indica dumort: Scientifically unexplored.

Reinwardtia indica (Lineceae) is a medicinal plant cultivated in the Himalayan region. It is effectively used in folk medicines for the treatment of various health complications. In the present study, the shade dried leaves and stem were extracted in three different solvents such as ethyl acetate, ethanol, and hydro-alcoholic. The antioxidant efficacy of these extracts was confirmed by using different in vitro assays: DPPH free radical scavenging, superoxide radical scavenging, lipid peroxidation, metal ion chelating capability and reducing power determination. Total phenol content was maximum in hydro alcoholic extract of leaf (540.37 mg per g of gallic acid equivalents) and stem (330.51 mg per g of gallic acid equivalents) while flavonoid content was maximum in ethanolic extract of leaf (305 mg per gram of rutin equivalents) and ethyl acetate extract of stem (170.6 mg per gram of rutin equivalents). The antioxidant activity of these extracts was positively correlated with their total phenol and flavonoid content. Among all tested extracts, ethanolic extract of leaf exhibit maximum zone of inhibition against all tested clinical isolates of bacterial (E. coli 11.00 ±â€¯1.73 mm, P. aeurogenosa 11.67 ±â€¯0.58 mm and S. aureus 10.33 ±â€¯1.53 mm) and fungal (C. albicans 11.33 ±â€¯1.10 mm) pathogens, while ethyl acetate extracts of the leaf and stem showed minimum inhibitory concentration against all tested microorganisms. Thus, R. indica extracts can be used as potent natural antioxidant having antifungal and antibacterial action.

Molecular characterization of dengue viruses circulating during 2009-2012 in Uttar Pradesh, India.

Dengue is the most rapidly spreading mosquito-borne viral disease in the world; in India it has taken endemic proportion implicating all the four known dengue virus serotypes. Dengue infection is caused by a small, single stranded RNA virus comprising of four antigenically distinct virus serotypes designated as dengue virus type 1-4 (DENV-1-4). On the basis of genomic variations, each serotype is classified further into its genotypes. Epidemiological studies have shown that the emergence of a newer dengue serotype/genotype after an interval always leads to a major outbreak; therefore a continuous epidemiological surveillance is needed to monitor the epidemiology of dengue viruses. The present study was planned to identify the serotype/genotype of dengue viruses circulating in Uttar Pradesh, India. Of 433 dengue suspected patients, tested by reverse transcriptase PCR (RT-PCR), 136 were positive for dengue virus RNA. Of these, DENV-1, 2, and 3 were detected in 26 (19.1%), 77 (56.6%), and 33 (24.3%) patients, respectively. Of 136 RT-PCR positive samples, 24 samples were sequenced to identify their genotypes. For sequencing C-prM gene junction of dengue virus genome was chosen. Phylogenetic analysis of sequenced dengue strains revealed that all the 12 DENV-1 strains were genotype III, all the eight DENV-2 strains were genotype IV (Cosmopolitan genotype) and among four DENV-3 strains, three were genotype III and one was genotype I. In conclusion, the co-circulation of multiple dengue virus serotypes and genotypes is alarming in U.P., India.

Serum levels of IL-8, IFNγ, IL-10, and TGF ß and their gene expression levels in severe and non-severe cases of dengue virus infection.

Increased serum and mRNA levels of cytokines in patients with dengue virus (DV) infection suggest that cytokines are one of the key factors in the pathogenesis of disease caused by this virus. Here, we tested 211 serum and 56 mRNA samples from an equal number of dengue cases to determine the levels of interleukin-8 (IL-8), interferon gamma (IFN-γ), interleukin-10 (IL-10) and transforming growth factor beta (TGF-ß). A total 70 serum and 15 mRNA samples from healthy individual were also tested for cytokines and served as controls. Serum and mRNA levels of IL-8 were highest in the earlier days of dengue infection. IFNγ levels peaked one or two days before defervescence. Levels of IL-10 and TGF-ß were highest later in dengue infection, and TGF-ß levels peaked on the day of defervescence. Mean levels of IFNγ, TGF ß and IL-10 were higher in samples from dengue cases, irrespective of severity, than in healthy controls. In contrast, the level of IL-8 was significantly higher in samples from severe dengue cases and lower in cases of dengue without warning signs than in healthy controls. Children (82.2 % of 101 paediatric cases) commonly had severe dengue illness. Samples that were positive for anti-DV IgG antibody had higher levels of IL-8 and TGF ß. DV-2 infections were associated with severe dengue illness. IL-8 and IFNγ levels were higher in the presence of warning signs of severe dengue. Levels of IL-8, IL-10 and TGF ß were independently associated with disease outcome. These data provide evidence of an association of IL-8, IFNγ, TGF ß and IL-10 levels with the severity of dengue illness. Especially, IL-8 levels can be used as a predictor of severe DV infection.

Techno-environmental analysis to valorize the secondary energy resources from refuse-derived fuel-based waste to energy plant.

The present study quantifies the environmental and sustainability impacts associated with municipal solid waste management (MSWM) in India which plays a vital environmental issue in recent times. The upsurge in population has resulted in massive waste generation, leading to a concerning rise in the level of greenhouse gas (GHG) emissions. Therefore, the sustainable management of MSW has been discussed and highlights the conversion of MSW into refuse-derived fuel (RDF) to identify its potential for generating electricity in waste-to-energy (WtE) plants. The life cycle assessment (LCA) study has been done to identify and compare the environmental impacts associated with different scenarios (SC) as SC1: landfilling without energy recovery, SC2: open burning and SC3: processing of RDF in WtE plant by considering the nine impact categories from the inventory data obtained over a period of 12 consecutive months (Jan 2021-Jan 2022). The results exhibited that the global warming potential caused by emissions of GHG are in the order of SC1 (1188 kg CO2 eq) > SC2 (752 kg CO2 eq) > SC3 (332 kg CO2 eq), respectively from 1 t of MSW. It is concluded that the WtE plant can help in the reduction of environmental issues, strengthening the capacity of electricity generation and improving the aesthetic view of the city which is socially acceptable as well. Thus, WtE technology can help in achieving sustainable development goal 12 to regenerate the sustainable secondary resources for the twenty-first century and minimize global climate change.

IL-17 level in patients with Dengue virus infection & its association with severity of illness.

BACKGROUND: Clinical symptoms of Dengue vary from mild febrile illness to severe infection. A potent pro-inflammatory cytokine, IL-17, secreted by mainly Th17 cells mediate inflammation and autoimmune diseases. Role of IL-17 in pathogenesis of dengue virus (DV) infection is not clear. METHODS: Total 211 dengue patients and 70 healthy controls were enrolled. IL-17 level was tested in serum samples from all cases and controls. Cases were grouped as either dengue or severe dengue; based on WHO (2009) classification. Anti DV IgG antibody detection and DV serotype specific PCR were also done. Levels of IL-17 in dengue and severe dengue patients, primary and secondary DV infection were compared. Association of DV serotypes with severity of illness and various clinical and laboratory features with IL-17 levels were analyzed. RESULTS: Of total 211cases, 90 and 121cases were presenting as dengue and severe dengue illness, respectively. Levels of IL-17 were significantly higher in dengue patients as compared to control. Mean level of IL-17 was higher in severe cases than non severe cases; however difference was not statistically significant. Stratified analysis of IL-17 level in different age group showed significantly high IL-17 levels in children with severe dengue. Mean level of IL-17 was also significantly higher in cases with secondary DV infection in comparison to primary DV infection. Levels of IL-17 were higher in patients with DV-2 infection in comparison to cases with DV-1 and DV-3 infection. Significant positive association of high IL-17 levels was seen with pleural effusion and respiratory distress. CONCLUSION: IL-17 appears to be associated with severe DV infection. It is very important to understand the exact role of IL-17 in the pathogenesis of severe DV infection.

A nanobody based ultrasensitive electrochemical biosensor for the detection of soluble CTLA-4 -A candidate biomarker for cancer development and progression.

A soluble isoform of cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) has been found in the serum of healthy individuals and alterations in its expression level have been linked with the development and progression of various cancers. Conventionally, soluble CTLA-4 (sCTLA-4) has been quantified by techniques such as ELISA, western blot, and flow cytometry, which however are time-consuming, highly expensive and require large sample volumes. Therefore, rapid, cost-effective and real-time monitoring of soluble CTLA-4 levels is much needed to facilitate timely diagnosis of a worsening disease and help patient selection for immunotherapeutic interventions in cancer. Here, for the first time, we report an ultrasensitive, highly selective electrochemical nanobody (NAb) based biosensor for the quantitative detection of soluble CTLA-4 employing poly(3,4-ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) and gold nanoparticles modified electrode with attomole sensitivity. Incorporating nanomaterials with conductive polymers enhances the sensitivity of the electrochemical biosensor, while the nanobody's stability, specificity and ease of production make it a suitable choice as a bioreceptor. The proposed NAb-based sensor can detect sCTLA-4 from pure recombinant protein in a wide concentration range of 100 ag mL-1- 500 µg mL-1, with a limit of detection of 1.19 ag mL-1 (+3σ of the blank signal). The sensor's relative standard deviation for reproducibility is less than 0.4% and has effective real sample analytics for cell culture supernatant with no significant difference with pure recombinant protein (p < 0.05). Our proposed nanobody based sensor exhibits stability for up to 2 weeks (<3% variation). Moreover, this nanobody-based sensor presents a future opportunity for quantitative, ultrasensitive, and economical biosensor development that can be adapted to monitor the immune landscape of cancer patients to provide a larger therapeutic window.

Trend of dengue virus infection at Lucknow, north India (2008- 2010): a hospital based study.

BACKGROUND & OBJECTIVES: Dengue virus (DV) infection has emerged as a major health problem in north India. Here, we report the annual trend of dengue virus infection as seen in Lucknow, Uttar Pradesh, during 2008-2010. METHODS: Blood samples from clinically suspected cases of dengue virus infection were collected and history was taken on structured clinical data sheet. All samples were tested for dengue IgM by antibody capture ELISA. Selected samples were tested by conventional RT-PCR for dengue virus RNA. Weather information was continuously recorded from website of world weather information service . RESULTS: There was a gradual increase in number of dengue fever cases with increased occurrence in 2010. Cases referred in January - December 2008 were 398 (54.5% anti DV IgM positive), in January - December 2009 were 599 (51.9% anti DV IgM positive) and in January - December 2010 were 1602 (64.9% anti DV IgM positive). Serotypes circulating in years 2008, 2009 and 2010 were DV-2 & DV-3, DV -1, 2 & 3 and DV-1 and DV-2 respectively. There is no statistical significant correlation between weather data and increasing dengue positive cases. INTERPRETATION & CONCLUSIONS: Increased cases of dengue fever were seen in 2010, which was not correlated with any change in environmental factors. A change in circulating serotypes was noted.

Effect of Nigella sativa seeds extracts on iNOS through antioxidant potential only: crude/total extract as molecular therapy drug.

There is general belief that only pure phytomolecules may be used as molecular therapeutic agent through one to one action. However, the traditional systems of medicine e.g. Ayurveda, uses the crude extracts, mostly water decoctions and oils, as drug. A comparative study of hexane, ethyl acetate and methanol fractions of N. Sativa seeds has been carried out on fresh rat-peritoneal-macrophage culture with reference to their role on various targets of lipopolysaccharide induced release of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) expression. The results indicated significant antioxidant potential with methanolic extract as most effective. Its mechanism of action was proposed primarily through its antioxidant potential and not through direct inhibition of other kinases, involved in its signaling cascade.

Outcomes of Posterior Approach Surgery in Various Types and Grades of Upper Eyelid Blepharoptosis in Indian Eyes.

PURPOSE: To present the outcomes of levator plication (levatorpexy) surgery by posterior approach for correction of ptosis of different aetiologies in Indian eyes. METHODS: Retrospective review of clinical notes and clinical photos of consecutive ptosis patients who underwent levatorpexy from January 2017 to September 2018. Surgery was considered successful if the following four criteria were simultaneously met: a postoperative MRD1 of ≥2 mm and ≤4.5 mm, inter eyelid height asymmetry of ≤1 mm, no overcorrection compared to the opposite eye, and a satisfactory eyelid contour determined by patient. RESULTS: Thirty six eyelids of 36 patients (mean age, 23.6 years; 25 females) underwent unilateral levatorpexy with the left eyelid affected in 66.6 %. Twenty one congenital (58.3%) (C), 12 (33.3%) aponeurotic (A) and 3 (8.3%) complex ptosis (CX) had a mean levator function of 9.8 mm (range, 6 to 15). Mean pre and post operative margin reflex distance (MRD 1) was 0.611mm (range, -1 to 3 mm) and 3 .00 mm (range, 2 to 4 mm) respectively. At the final follow up of 12 months, four patients remained under corrected (11.1%). At the final follow up of twelve months, the success rate was 78 percent. CONCLUSION: Posterior approach for ptosis repair offers a scar less, minimally invasive, easy to revise, and successful lid height correction in congenital and acquired ptosis.

Vibration-based biomimetic odor classification.

Olfaction is not as well-understood as vision or audition, nor technologically addressed. Here, Chemical Graph Theory is shown to connect the vibrational spectrum of an odorant molecule, invoked in the Vibration Theory of Olfaction, to its structure, which is germane to the orthodox Shape Theory. Atomistic simulations yield the Eigen-VAlue (EVA) vibrational pseudo-spectra for 20 odorant molecules grouped into 6 different 'perceptual' classes by odour. The EVA is decomposed into peaks corresponding to different types of vibrational modes. A novel secondary pseudo-spectrum, informed by this physical insight-the Peak-Decomposed EVA (PD-EVA)-has been proposed here. Unsupervised Machine Learning (spectral clustering), applied to the PD-EVA, clusters the odours into different 'physical' (vibrational) classes that match the 'perceptual', and also reveal inherent perceptual subclasses. This establishes a physical basis for vibration-based odour classification, harmonizes the Shape and Vibration theories, and points to vibration-based sensing as a promising path towards a biomimetic electronic nose.

LungINFseg: Segmenting COVID-19 Infected Regions in Lung CT Images Based on a Receptive-Field-Aware Deep Learning Framework.

COVID-19 is a fast-growing disease all over the world, but facilities in the hospitals are restricted. Due to unavailability of an appropriate vaccine or medicine, early identification of patients suspected to have COVID-19 plays an important role in limiting the extent of disease. Lung computed tomography (CT) imaging is an alternative to the RT-PCR test for diagnosing COVID-19. Manual segmentation of lung CT images is time consuming and has several challenges, such as the high disparities in texture, size, and location of infections. Patchy ground-glass and consolidations, along with pathological changes, limit the accuracy of the existing deep learning-based CT slices segmentation methods. To cope with these issues, in this paper we propose a fully automated and efficient deep learning-based method, called LungINFseg, to segment the COVID-19 infections in lung CT images. Specifically, we propose the receptive-field-aware (RFA) module that can enlarge the receptive field of the segmentation models and increase the learning ability of the model without information loss. RFA includes convolution layers to extract COVID-19 features, dilated convolution consolidated with learnable parallel-group convolution to enlarge the receptive field, frequency domain features obtained by discrete wavelet transform, which also enlarges the receptive field, and an attention mechanism to promote COVID-19-related features. Large receptive fields could help deep learning models to learn contextual information and COVID-19 infection-related features that yield accurate segmentation results. In our experiments, we used a total of 1800+ annotated CT slices to build and test LungINFseg. We also compared LungINFseg with 13 state-of-the-art deep learning-based segmentation methods to demonstrate its effectiveness. LungINFseg achieved a dice score of 80.34% and an intersection-over-union (IoU) score of 68.77%-higher than the ones of the other 13 segmentation methods. Specifically, the dice and IoU scores of LungINFseg were 10% better than those of the popular biomedical segmentation method U-Net.

Anacardic acid encapsulated solid lipid nanoparticles for Staphylococcus aureus biofilm therapy: chitosan and DNase coating improves antimicrobial activity.

Biofilm mediated bacterial infections are the key factors in the progression of infectious diseases due to the evolution of antimicrobial resistance. Traditional therapy involving antibiotics is not adequate enough for treatment of such infections due to the increased resistance triggered by biofilm. To overcome this challenge, we developed anacardic acid (Ana) loaded solid lipid nanoparticles (SLNs), further coated with chitosan and DNase (Ana-SLNs-CH-DNase). The DNase coating was hypothesized to degrade the e-DNA, while chitosan was coated to yield positively charged SLNs with additional adhesion to biofilms. The SLNs were developed using homogenization method and further evaluated for particle size, polydispersity index, zeta potential, and entrapment efficiency. Drug excipient compatibility was confirmed by using FT-IR study, while encapsulation of Ana in SLNs was confirmed by X-ray diffraction study. The SLNs demonstrated sustained release for up to 24 h and excellent stability at room temperature for up to 3 months. The developed SLNs were found non-toxic against human immortalized keratinocyte (HaCaT) cells while demonstrated remarkably higher antimicrobial efficacy against Staphylococcus aureus. Excellent effect of the developed SLNs on minimum biofilm inhibition concentration and minimum biofilm eradication concentration further confirmed the superiority of the developed formulation strategy. A significant (p < 0.05) reduction in biofilm thickness and biomass, as confirmed by confocal laser scanning microscopy, was observed in the case of developed SLNs in comparison with control. Cumulatively, the results suggest the enhanced efficacy of the developed formulation strategy to overcome the biofilm-mediated antimicrobial resistance. Graphical abstract.

Effect of hexane fraction of leaves of Cinnamomum tamala Linn on macrophage functions.

The leaves of Cinnamomum tamala Linn (Lauraceae), component of Indian spices are associated with hypoglycemic property in Ayurveda; however, no report is available towards its immunomodulation property, which has been explored here. The dried powder of CT leaves was extracted with hexane and solvent free extract (CTH) was given orally to rats for 10 days, in various doses. Its effect was studied on peritoneal macrophage functions, and was compared with ascorbic acid (1,000 mg/kg, immune-stimulant) and cyclophosphamide (10 mg/kg, immune-suppressant). CTH significantly suppressed phagocytosis activity (EC(50) 2,355 +/- 52.45 mg/kg), reduced production of superoxide (EC(50) 275.91 +/- 10.21 microg/ml) and cellular NADPH (EC(50) 384.959 +/- 4.85 microg/ml) content in concentration dependent manner. It also inhibited LPS induced production of nitric oxide (EC(50) 143.75 +/- 3.40 microg/ml) and iNOS protein expression (EC(50) 183.132 microg/ml). Thus, it could be suggested that non-polar hexane fraction of leaves of C. tamala possesses immunosuppressive property, which is mediated through modulation of innate immunity.

Oily fraction of Semecarpus anacardium Linn nuts involves protein kinase C activation for its pro-inflammatory response.

The oily fraction (non polar fraction-NPF) of S. anacardium (SA) significantly increased the expression of protein kinase C-delta (PKC-delta) in macrophages in concentration dependent manner, which was similar to phorbol myristate acetate (PMA) response. Further, H-7 (1-(5-isoquinolinesulphonyl)-2-methylpiperazine), an inhibitor of PKC significantly inhibited this NPF mediated response in a concentration dependent manner. In the post treatment kinetics, H-7 showed this inhibition only up to 6 min post NPF/PMA addition, but in similar condition, quercetin, a flavone with reported antioxidant property, showed this inhibition only up to 2 min. The results clearly suggest that oily fraction of SA nuts enhances the expression of PKC protein, which may be responsible for its reported pro-inflammatory property.

Overexpression of efflux pump transporter genes and mutations in ERG11 pave the way to fluconazole resistance in Candida tropicalis: A study from a North India region.

INTRODUCTION: Fluconazole resistance in Candida tropicalis health care-associated infections is increasing. We investigated the role of efflux pump and mutations in ERG11p conferring fluconazole resistance in C. tropicalis. MATERIALS AND METHODS: A total of 17 C. tropicalis clinical isolates, including fluconazole-resistant and fluconazole-susceptible/fluconazole-susceptible dose-dependent isolates, were collected from a tertiary care centre in a North India region between 2015 and 2018. Antifungal susceptibility, reversal of fluconazole resistance by tacrolimus, ERG11 amplification and its sequencing and a quantitative polymerase chain reaction (PCR) assay for expression analysis of ERG11, MDR1 and CDR1 genes were performed. RESULTS AND DISCUSSION: Synergism between fluconazole and tacrolimus was observed in all resistant C. tropicalis isolates. Overexpression of all the three genes, MDR1, ERG11 and CDR1, was observed in resistant isolates (P = 0.05). Among resistant isolates, mutations leading to amino acid substitution were seen in two, Ct10 (glysine464serine) and Ct16 (tyrosine132phenylalanine; serine154phenylalanine). CONCLUSION: Overexpression in efflux pump transporter genes, together with mutations in ERG11, lead to fluconazole resistance among C. tropicalis. To the best of our knowledge, this is the first study on the C. tropicalis fluconazole resistance mechanism from the North India region.