Evaluation of Safety and Potency of Kyasanur Forest Disease (KFD) Vaccine Inactivated with Different Concentrations of Formalin and Comparative Evaluation of In Vitro and In Vivo Methods of Virus Titration in KFD Vaccine.

We evaluated the safety and potency of the Kyasanur Forest disease (KFD) vaccine inactivated with different formalin concentrations in mice, since the side effects due to higher formalin concentrations have been a major reason for vaccine refusal. Furthermore, with an objective to reduce the use of mice in vaccine testing, we performed quantification of the KFD virus by real-time PCR and compared it with in vivo titration in mice. The KFD vaccine prepared in chicken embryo fibroblast cells was inactivated with 0.04%, 0.06%, and 0.08% concentrations of formalin. The vaccine inactivated with 0.04% and 0.06% formalin failed the safety test, whereas the KFD vaccine inactivated with 0.08% formalin was safe and potent with a log protective index of 5678 in mice. This reduced formalin content may induce no/lesser side-effects of pain/swelling which may increase the vaccine acceptance. The real-time PCR on individual KFD vaccine harvests interpreted that when the CT value of each harvest is <20, the vaccine will have sufficient viral particles to pass the potency test. Comparison of the real-time PCR on tenfold dilutions of the pooled harvests with in vivo mice inoculation test revealed that the 1MLD50 of the vaccine lies in the tenfold dilution that yields CT values between 31 and 34.

Astaxanthin and DHA supplementation ameliorates the proteomic profile of perinatal undernutrition-induced adipose tissue dysfunction in adult life.

Maternal diet is an essential factor that directly and indirectly regulates fetal growth. Exposure to certain environmental conditions substantially impacts an individual's short- and long-term health. Adipose tissue dysfunction is a worldwide chronic disease caused by improper lipid build-up in adipose tissue leading to obesity. Therefore, it is the need of the hour to invent anti-obesity agents. As a keto-carotenoid, Astaxanthin (AsX) has been shown to have preventive effects against problems associated with obesity. A crucial role in the pathogenesis of obesity has been attributed to dietary polyunsaturated fatty acids. Adipose tissue plays a vital role in maintaining overall body homeostasis. Metabolic dysfunction of white adipocytes forms a critical step in the emergence of insulin resistance and related diseases. Here we aim to investigate the effect of AsX and Docosahexaenoic acid (DHA) supplementation on the proteomic profile of perinatal undernutrition-induced adipose tissue dysfunction in adult life using a rat model. The LC-MS/MS quantitative proteomics enabled us to identify differentially expressed proteins in perinatal undernourished but AsX and DHA-supplemented animal models. Data are available via ProteomeXchange with identifier PXD041772.This study explored biological roles, molecular functions of differentially expressed proteins, and pathways related to adipose tissue dysfunction induced by undernutrition and its effective modulation by AsX and DHA.

Nutritional efficacy of Astaxanthin in modulating orexin peptides and fatty acid level during adult life of rats exposed to perinatal undernutrition stress.

Perinatal undernutrition stress predisposes several disorders in adult life, which could be programed using nutraceuticals. However, the effect of perinatal undernutrition stress on orexin peptides, brain lipids, and its amelioration by a potent antioxidant (Astaxanthin) needs exploration. The present study focussed on the effect of perinatal undernutrition stress on brain fatty acid levels, Orexin peptides A and B, and its amelioration by Astaxanthin.Twenty-four male Wistar rats (Rattus norvegicus) were allocated to four groups (n = 6) as Normal, Perinatally Undernourished (UN), Astaxanthin treated (AsX, 12mg/kg), and perinatally Undernourished-but-Astaxanthin treated (UNA), and are allowed to grow for 1, 6 and 12 months. The fatty acid and orexin peptides A & B at different brain parts were measured and compared. Orexin peptides were assessed using an ELISA kit. Fatty acid levels were estimated using HP 5890 gas chromatograph. Data were analyzed by ANOVA followed by Tukey's posthoc test. P < 0.05 was considered significant.The hair cortisol, Orexin-A, and B were significantly increased (p < 0.001) in the UN group compared to normal and were modulated significantly by AsX in the UNA group. Undernutrition stress during the perinatal period altered the lipid profile, Total SFA, Total MUFA, Total n-3 PUFA, Total n-6 PUFA, n-3: n-6 PUFA, which Astaxanthin effectively modulated at 6 and 12 months of postnatal life. There was no difference between DHA and AA ratio. These results indicate that nutritional enrichment with Astaxanthin during the perinatal period positively contributes to adult health. Further, the mechanism of regulation of brain chemistry by Astaxanthin is warranted.

Role of astaxanthin in the modulation of brain-derived neurotrophic factor and spatial learning behavior in perinatally undernourished Wistar rats.

Objective: Maternal health and nutrition during the perinatal period is the predominant factor influencing the functional development of the brain. Maternal malnutrition during the perinatal period causes retardation of brain development. The current study investigates the role of Astaxanthin (AsX) in spatial learning and memory and BDNF in perinatally undernourished Wistar rats.Methods: The albino wistar rats were perinatally undernourished and administered with different dosages of AsX. The spatial learning and memory performance and BDNF level were assessed. Data were collected and analysed.Results: The % Correct choice during the acquisition phase, performance at the end of the acquisition phase and the mean BDNF level at the Hippocampus, Cerebellum, and Cerebral cortex showed significant decline (P<0.001) in the PUN group and significantly high (P<0.001) in the PUNA2 group compared to the control. However, the mean RME and mean WME during different days of the acquisition phase were significantly high (P<0.001) in the PUN group and insignificant (P>0.05) in PUNA2 compared to the control.Discussion: The results showed that AsX effectively modulated the cognitive deficit that occurred in perinatally undernourished rats. This can be attributed to BDNF upregulation as evidenced by the significant increase of the BDNF level.

Structural and functional insights into phosphomannose isomerase: the role of zinc and catalytic residues.

Phosphomannose isomerase (PMI) is a housekeeping enzyme that is found in organisms ranging from bacteria to fungi to mammals and is important for cell-wall synthesis, viability and signalling. PMI is a zinc-dependent enzyme that catalyses the reversible isomerization between mannose 6-phosphate (M6P) and fructose 6-phosphate (F6P), presumably via the formation of a cis-enediol intermediate. The reaction is hypothesized to involve ring opening of M6P, the transfer of a proton from the C2 atom to the C1 atom and between the O1 and O2 atoms of the substrate, followed by ring closure resulting in the product F6P. Several attempts have been made to decipher the role of zinc ions and various residues in the catalytic function of PMI. However, there is no consensus on the catalytic base and the mechanism of the reaction catalyzed by the enzyme. In the present study, based on the structure of PMI from Salmonella typhimurium, site-directed mutagenesis targeting residues close to the bound metal ion and activity studies on the mutants, zinc ions were shown to be crucial for substrate binding. These studies also suggest Lys86 as the most probable catalytic base abstracting the proton in the isomerization reaction. Plausible roles for the highly conserved residues Lys132 and Arg274 could also be discerned based on comparison of the crystal structures of wild-type and mutant PMIs. PMIs from prokaryotes possess a low sequence identity to the human enzyme, ranging between 30% and 40%. Since PMI is important for the virulence of many pathogenic organisms, the identification of catalytically important residues will facilitate its use as a potential antimicrobial drug target.

Effects of zinc oxide nanoparticles and zinc sulfate on the testis of common carp, Cyprinus carpio.

The present study analyzed the effects of zinc oxide nanoparticles (ZnO-NPs) and zinc sulfate (ZnSO4) in the testis of six-month-old common carp Cyprinus carpio exposed to three different doses, viz., 10, 50, and 100 µg/L for 21 days. Characterization of ZnO-NPs was done after sonication, the size and shape of ZnO-NPs were determined as ∼20-30 nm spherical structure measured zeta potential of +26.0 mV. After treatment, determination of zinc (Zn) concentration in the testes revealed desired impact of the exposure. Expression of several transcription factors and few steroidogenic enzyme genes in the treated testis showed significant downregulation than the control. Measurement of oxidative stress-related enzymes such as catalase, superoxide dismutase, and glutathione-S-transferase revealed substantial elevation in the testis of treated groups when compared to control. Histological analysis of testis exhibited dose-related response, defective lumen, and slow progression of spermatogenesis. Exposure of both the forms of Zn on TM3 Leydig cell culture displayed loss of adhesion, clumping with decreased viability, and a significant increase in the apoptotic cells. In addition, comet and intracellular reactive oxygen species (ROS) assays authenticated DNA damage upon treatment with a significant increase in ROS. Histological analysis after treatment withdrawal showed revival of testis in carp to rescue the effect. Thus, the present report highlights the adverse effect of Zn on the testis function in common carp as well as evident drastically toxic in in vitro cultures.

Anti-aging Role of Curcumin by Modulating the Inflammatory Markers in Albino Wistar Rats.

PURPOSE: Advanced age is associated with an accumulation of free radical damage, which leads to physiological and clinical modifications. Numerous pharmaceutical and nutraceuticals are considered to influence longevity and prompting healthy ageing. Therefore, the current study attempted to investigate Curcumin's role in the inflammatory indices as anti-ageing marker in albino Wistar rats. METHODS: Twelve months old rats were used in the study, grouped as Normal control (NC), Sham control (SC), Curcumin-1, Curcumin-2 and Curcumin-3. Last three groups received Curcumin at the dosages of 100 mg, 200 mg and 400 mg/kg body weight respectively. After six months of intervention, blood was collected for the estimation of C-reactive protein (CRP), Serum Albumin, Globulin, Lymphocyte percentage, Total Antioxidant Capacity (TAC), Malondialdehyde (MDA), Superoxide Dismutase (SOD) and Nitric Oxide (NO) level using standard procedures. RESULTS: There was a significant decline in the CRP level (p < 0.05) in rats treated with 200 mg and 400 mg of Curcumin/kg body weight. The MDA level was found to be significantly increased (p < 0.05) in animals fed with 400 mg of Curcumin/kg body weight as compared to NC. The NO level was increased significantly (p < 0.05) in rats treated with 200 and 400 mg of Curcumin/kg body weight. CONCLUSION: Finding of the study suggests that Curcumin exhibits favorable influence in slowing down of ageing process by suppressing age-related changes in inflammatory indices.

Study of Effect of Magnesium Sulphate in Management of Acute Organophosphorous Pesticide Poisoning.

BACKGROUND: Organophosphorus compound poisoning (OPCP) is a major public health problem in developing countries like India. Atropine and oximes remain the main-stay of management. Magnesium sulfate (MgSO4) has shown benefit in the management of OPCP. AIMS: This study was designed to assess the effect of MgSO4 on outcome in OPCP patients admitted to Intensive Care Unit (ICU). SETTINGS AND DESIGN: Double-blind prospective randomized clinical trial in an ICU of tertiary care institution. METHODS: One hundred patients (50 in each group) of OPCP, confirmed by history and syndrome of OPCP with low plasma pseudocholinesterase, aged between 18 and 60 years were studied. Magnesium group (Group M) received 4 g of 20% MgSO4 infusion over 30 min at admission to ICU, control group (Group C) received normal saline placebo in the same manner. Patients were assessed for the need for intubation, requirement of atropine, duration of mechanical ventilation, duration of ICU stay, and its effect on mortality. STATISTICAL ANALYSIS: Chi-square test and Fisher's exact test for categorical data, independent sample t-test, and paired t-test for nominal data. RESULTS: Demographics and basal serum magnesium levels were comparable. Atropine requirement was higher in Group C (74.82 ± 22.39 mg) compared to Group M (53.11 ± 45.83 mg) (P < 0.001). A total of 33 patients in Group C and 23 patients in Group M required intubation, respectively (P = 0.043). The mean duration of mechanical ventilation was 4.51 ± 2 days in Group C compared to 4.13 ± 1.6 days in Group M (P = 0.45). ICU stay was 5.36 ± 2.018 days in Group C compared to 4.54 ± 1.581 days in Group M (P = 0.026). There was no significant difference in mortality between the groups. CONCLUSION: Four grams of MgSO4 given to OPCP patients within 24 h of admission to ICU, decreases atropine requirement, need for intubation, and ICU stay.

Antiviral effects of black raspberry (Rubus coreanus) seed extract and its polyphenolic compounds on norovirus surrogates.

Black raspberry seeds, a byproduct of wine and juice production, contain large quantities of polyphenolic compounds. The antiviral effects of black raspberry seed extract (RCS) and its fraction with molecular weight less than 1 kDa (RCS-F1) were examined against food-borne viral surrogates, murine norovirus-1 (MNV-1) and feline calicivirus-F9 (FCV-F9). The maximal antiviral effect was achieved when RCS or RCS-F1 was added simultaneously to cells with MNV-1 or FCV-F9, reaching complete inhibition at 0.1-1 mg/mL. Transmission electron microscopy (TEM) images showed enlarged viral capsids or disruption (from 35 nm to up to 100 nm) by RCS-F1. Our results thus suggest that RCS-F1 can interfere with the attachment of viral surface protein to host cells. Further, two polyphenolic compounds derived from RCS-F1, cyanidin-3-glucoside (C3G) and gallic acid, identified by liquid chromatography-tandem mass spectrometry, showed inhibitory effects against the viruses. C3G was suggested to bind to MNV-1 RNA polymerase and to enlarge viral capsids using differential scanning fluorimetry and TEM, respectively.

Plasma exchange in Immunoglobulin A nephropathy with thrombotic microangiopathy and acute cortical necrosis.

A 25-year-old female presented with decreased urine output, deranged renal function, thrombocytopenia, and hemolytic anemia. Kidney biopsy was consistent with thrombotic microangiopathy with acute cortical necrosis and Immunoglobulin A nephropathy (IgAN). Hemolytic anemia, thrombocytopenia and urine output improved after five sessions of plasma exchange. Renal function showed a delayed recovery and serum creatinine normalized by 3 months. This is first case of successful use of plasma exchange in hemolytic uremic syndrome with cortical necrosis associated with IgAN.

Proliferative glomerulonephritis with monoclonal immunoglobulin deposition disease: The utility of routine staining with immunoglobulin light chains.

Proliferative glomerulonephritis occurring as a consequence of monoclonal glomerular deposits of IgG is uncommon. It is a form of renal involvement in monoclonal gammopathy that mimics immune complex glomerulonephritis. Here, we report the first series of proliferative glomerulonephritis with monoclonal IgG deposits (PGNMID) from the Indian subcontinent highlighting use of light chain immunofluorescence (IF) in routine renal biopsy interpretation. We retrieved 6 patients diagnosed as proliferative glomerulonephritis with monoclonal IgG deposits (PGNMID) out of 160 biopsies (3.7%) with membranoproliferative patterns over 5 1/2 years (2009-2014), one of whom had recurrence 6 months post-renal transplant. Four (67%) patients presented with rapidly progressive renal failure and two (33%) with nephrotic syndrome. None of these patients had overt multiple myeloma. The predominant histologic pattern was membranoproliferative with all the biopsies showing IgG3 Kappa deposits on IF. The deposits were primarily subendothelial on electron microscopy.

Crystal structure of 7,8-benzocoumarin-4-acetic acid.

The fused-ring system in the title compound [systematic name: 2-(2-oxo-2H-benzo[h]chromen-4-yl)acetic acid], C15H10O4, is almost planar (r.m.s. deviation = 0.031 Å) and the Car-C-C=O (ar = aromatic) torsion angle for the side chain is -134.4 (3)°. In the crystal, mol-ecules are linked by O-H⋯O hydrogen bonds, generating [100] C(8) chains, where the acceptor atom is the exocyclic O atom of the fused-ring system. The packing is consolidated by a very weak C-H⋯O hydrogen bond to the same acceptor atom. Together, these inter-actions lead to undulating (001) layers in the crystal.

Light chain proximal tubulopathy with cast nephropathy in a case of multiple myeloma.

The renal diseases most frequently associated with myeloma include cast nephropathy (CN), amyloidosis and monoclonal immunoglobulin deposition disease. Light chain proximal tubulopathy (LCPT) is reported less frequently. Majority of the cases with κ-restriction present with Fanconi syndrome (FS) and show crystals in proximal tubular epithelial cytoplasm. In contrast, those with λ-restriction are infrequently associated with FS and show cytoplasmic vacuolations in proximal tubular epithelial cytoplasm. Combination of morphologies in kidney affected by plasma cell dyscrasias is rare and co-existence of LCPT and CN is one of the rarest. We report a case of multiple myeloma having this rare combination of morphologies.

Successful induction of granulomatosis with polyangiitis with tacrolimus.

We report a 50-year-old female who presented with inflammatory arthritis, upper respiratory tract symptoms, and microscopic hematuria with nephrotic range proteinuria. Antineutrophil cytoplasmic antibodies (ANCA) were detectable and kidney biopsy showed pauci-immune focal necrotizing crescentic glomerulonephritis. She was treated with pulse intravenous cyclophosphamide (CYC) and prednisolone. Patient developed severe leucopenia after the first dose and subsequently had leucopenia to low dose CYC, mycophenolate mofetil and azathioprine were also tried. However, patient developed leukopenia with all the above agents. Initiation of tacrolimus (TAC) was followed by dramatic response: Proteinuria decreased, serum albumin normalized and C-ANCA and anti-PR3 ANCA assays became negative. This is the first successful case of TAC as an induction agent in a patient with GPA (ANCA associated vasculitis with renal involvement).

Monoclonal gammopathy associated membranous glomerulonephritis: A rare entity.

A 40-year-old male presented with nephrotic syndrome. Light microscopic analysis of the renal biopsy showed thickening of the glomerular capillary wall. Immunofluorescence examination revealed granular deposition of monoclonal immunoglobulin (Ig) G3-kappa and complement C3 along the glomerular basement membrane. Electron microscopy showed subepithelial electron dense deposits, thus confirming membranous glomerulonephritis (MGN) with monoclonal gammopathy. MGN with monoclonal gammopathy is an extremely rare but distinctive entity. This patient was treated with a combination of bortezomib, thalidomide and dexamethasone and showed partial remission of his nephrotic state and dysproteinemia.

Crystal structure of 2-(5-meth-oxy-1-benzo-furan-3-yl)acetic acid.

The benzo-furan residue in the title compound, C11H10O4, is essentially planar (the r.m.s. deviation for the nine non-H atoms = 0.011 Å). While the meth-oxy group is coplanar with the fused ring system [C-C-O-C torsion angle = 3.1 (3)°], the acetic acid residue occupies a position almost prime [C-C-C-C = 77.0 (2)°]. In the crystal, centrosymmetrically related mol-ecules are linked by O-H⋯O hydrogen bonds to form eight-membered {⋯HOCO}2 synthons. The dimeric aggregates assemble into supra-molecular layers in the ab plane via benzene-C-H⋯O(ring) inter-actions.

Synthesis, DNA interactions and antibacterial PDT of Cu(II) complexes of phenanthroline based photosensitizers via singlet oxygen generation.

Cu(II) complexes [Cu(mqt)(B)H2O]ClO4(1-3) of 2-thiol 4-methylquinoline and phenanthroline bases (B), viz 1,10-phenanthroline (phen in 1), Dipyrido[3,2-d:2',3'-f]quinoxaline (dpq in 2) and Dipyrido[3,2-a:2',3'-c]phenazine (dppz in 3) have been prepared and characterized by elemental analysis, IR, UV-Vis, magnetic moment values, EPR spectra and conductivity measurements. The spectral data reveal that all the complexes exhibit square-pyramidal geometry. The DNA-binding behaviors of the three complexes were investigated by absorption spectra, viscosity measurements and thermal denaturation studies. The DNA binding constants for complexes (1), (2) and (3) were determined to 2.2×10(3), 1.3×10(4) and 8.6×10(4)M(-1) respectively. The experimental results suggest that these complexes interact with DNA through groove-binding mode. The photo induced cleavage studies shows that the complexes possess photonuclease property against pUC19 DNA under UV-Visible irradiation via a mechanistic pathway involving formation of singlet oxygen as the reactive species. Antimicrobial photodynamic therapy was studied using photodynamic antimicrobial chemotherapy (PACT) assay against Escherichiacoli and all complexes exhibited significant reduction in bacterial growth on photoirradiation.