Kynurenine pathway alteration in acute pancreatitis and its role as a biomarker of infected necrosis.

INTRODUCTION: Infected pancreatic necrosis (IPN) is a major cause of mortality in acute pancreatitis (AP). Currently, no specific strategies are available to predict the development of IPN. Earlier we reported that persistent down-regulation of HLA-DR increases risk of developing IPN. Altered kynurenine pathway (KP) metabolites showed poor prognosis in sepsis. Here we evaluated the role of HLA-DR and KP in IPN. METHODS: Patients with ANP and healthy controls were enrolled. Demographic and clinical parameters were recorded. Circulating interleukin (IL)-8, 6, 1ß, 10, Tumor necrosis factor-α were quantified using flowcytometry. Plasma procalcitonin, endotoxin, and KP (tryptophan, kynurenine) concentrations were estimated using ELISA. qRT-PCR was conducted to evaluate mRNA expression of HLA-DR, IL-10, Toll like receptor-4 (TLR-4), and kynurenine-3-monooxygenase (KMO) genes on peripheral blood mononuclear cells. Plasma metabolites were quantified using gas chromatography mass spectrometry (GC-MS/MS). Standard statistical methods were used to compare study groups. Metaboanalyst was used to analyse/visualize the metabolomics data. RESULTS: We recruited 56 patients in Cohort-1 (IPN:26,Non-IPN:30), 78 in Cohort-2 (IPN:57,Non-IPN:21), 26 healthy controls. Increased cytokines, endotoxin, and procalcitonin were observed in patients with IPN compared to Non-IPN. HLA-DR and KMO gene expressions were significantly down-regulated in IPN groups, showed positive correlation with one another but negatively correlated with IL-6 and endotoxin concentrations. Increased IDO and decreased plasma tryptophan were observed in IPN patients. Metabolome analysis showed significant reduction in several essential amino acids including tryptophan in IPN patients. Tryptophan, at a concentration of 9 mg/ml showed an AUC of 91.9 (95%CI 86.5-97.4) in discriminating IPN. CONCLUSION: HLA-DR downregulation and KP alteration are related to IPN. The KP metabolite plasma tryptophan can act as a potential biomarker for IPN.

Immuno-analytical method development for detection of transgenic Cry1Ac protein and its validation.

BACKGROUND: Bacillus thuringiensis (Bt) synthesizes Cry1Ac protein, which is toxic to many lepidopteran pests, and the cry1ac gene has been expressed in several transgenic crop plants. The Cry1Ac protein has been isolated from Bt kurstaki HD73 and purified to homogeneity. Polyclonal antibodies were raised against purified Cry1Ac in rabbits and goat. Sandwich ELISA was developed for Cry1Ac using goat IgG as a coating antibody, and affinity-purified rabbit IgG as the primary antibody. RESULTS: The sensitivity of the assay was in the range of 0.47-1000 ng. It was subsequently employed in validating biological samples. Fifteen different cotton-seed samples were screened: 12 were found to be Bt positive and 3 Bt negative. The CS7 seeds showed the highest Bt content of 8.51 ± 0.45 µg g-1 , followed by CS8 (6.0 ± 0.02 µg g-1 ), CS15 (5.9 ± 0.03 µg g-1 ), CS9 (5.5 ± 0.05 µg g-1 ), and CS10 (4.83 ± 0.013 µg g-1 ). The CS5 seeds showed Bt content of 3.6 ± 0.21 µg g-1 . The F2 generation, CS6 (Kaveri seeds) showed lower Bt content (2.9 ± 0.06 µg g-1 ). The CL5 samples showed Cry1Ac content of 0.99 ± 0.009 µg g-1 . The amount of Cry1Ac protein in leaves, stem, and roots of germinated Bt cotton plants (CS10 and CS4) were 1.76 ± 0.15 µg g-1 , 1.9 ± 0.01 µg g-1 , 2.0 ± 0.1 µg g-1 , and 1.6 ± 0.15 µg g-1 , 1.9 ± 0.01 µg g-1 , and 2.0 ± 0.01 µg g-1 dry tissue, respectively. CONCLUSION: The method developed can be used for screening the expression levels of Cry1Ac in different transgenic Bt cultivars and also spurious Bt cotton seeds procured by farmers. © 2019 Society of Chemical Industry.

Proximate, mineral composition and antioxidant activity of traditional small millets cultivated and consumed in Rayalaseema region of south India.

BACKGROUND: Millets are a diverse group of small seeded grasses, widely grown around the world as cereal foods. This communication details the proximate, mineral profile and antioxidant activity of six different small millets (Finger, Foxtail, Proso, Little, Barnyard and Kodo millets) and their 21 cultivars that are traditionally cultivated and consumed in the region of Ralayaseema, south India. RESULTS: The proximate analysis revealed that these millets are rich in protein, fat, ash (mineral), total dietary fibre and total phenols with appreciable antioxidant activity. However, starch and amylose content was comparatively lower as compared to major millet sorghum. ICP-MS analysis of small millets demonstrated that they are rich in minerals such as Ca, P, K, Mg, Fe, Cu, Zn, Mn, Cr, Mo and Se. CONCLUSION: Finger and kodo millets were found to be nutritionally superior over other small millets. The results suggest that small millets have a potential to provide food security and can combat micronutrient malnutrition. © 2017 Society of Chemical Industry.

Bimetallic nanocomposite (Ag-Au, Ag-Pd, Au-Pd) synthesis using gum kondagogu a natural biopolymer and their catalytic potentials in the degradation of 4-nitrophenol.

Bimetallic nanoparticles (BNPs) constitute two different metal elements and exhibit relatively superior mechanistic and catalytic efficacies owing to their synergistic functions over monometallic nanoparticles. In the present study various bimetallic Ag-Au, Ag-Pd, Au-Pd nanoparticles were synthesized using a natural biopolymer gum kondagogu (GK) as a reducing and capping agent, by a simple and cost-effective method. The synthesized BNPs when characterized using UV-vis spectroscopy revealed a specific surface plasmon resonance band (SPR) of each nanocomposite. The average particle size of Ag-Au, Ag-Pd, and Au-Pd BNPs was found to be 23 ± 10.3, 21 ± 7.6, and 23 ± 9.4 nm respectively based on transmission electron microscopy analysis. Surface morphology and functional groups on the gum matrix of GK-BNPs were analyzed by XRD and FT-IR respectively. The bimetallic nanocomposites were evaluated for their catalytic reduction of 4-nitrophenol (4-NP) to 4-aminophenol in the presence of NaBH4. The kinetic studies performed, depicted rate constants for Ag-Au, Ag-Pd, and Au-PdNPs as 0.31, 0.39, and 0.28 min-1 respectively. The catalytic efficiencies of three bimetallic nanocomposites were of the following order Ag-Pd > Ag-Au > Au-Pd. This study establishes the catalytic potentials of the three different bimetallic nanocomposites in the reduction of 4-NP an environmental pollutant, and the impact of their synergistic property.

Purification and characterization of mycoferritin from Fusarium verticillioides MRC 826.

The fungus Fusarium verticillioides MRC 826 (ascomycetes species), a toxigenic isolate is capable of synthesizing mycoferritin only upon induction with iron in yeast extract sucrose medium. The molecular mass, yield, iron and carbohydrate contents of the purified mycoferritin were 460 kDa, 0.010 mg/g of wet mycelia, 1.0 and 40.2%, respectively. Native gel electrophoresis of the mycoferritin revealed two bands possibly representing isoforms of ferritin. Subunit analysis by SDS-PAGE showed a single protein subunit of ~24 kDa suggesting similar sized subunits in the structure of apoferritin shell. Immunological cross reactivity was observed with the anti-fish liver ferritin. Transmission electron microscopy revealed an apparent particle size of 100 Å. N-terminal amino acid sequencing of mycoferritin showed identities with other eukaryotic ferritin sequences. The spectral characteristics were similar to equine spleen ferritin. However, circular dichroic spectra revealed a higher degree of helicity. Functionally, induction of mycoferritin minimizes the prooxidant role of iron.

Synthesis, characterisation and anti-tumour activity of biopolymer based platinum nanoparticles and 5-fluorouracil loaded platinum nanoparticles.

A facile and green synthesis of platinum nanoparticles [gum kondagogu platinum nanoparticles (GKPtNP)] using biopolymer- gum kondagogu was developed. The formation of GKPtNP was confirmed by ultraviolet (UV)-visible spectroscopy, scanning electron microscopy-energy dispersive X-ray spectroscopy, transmission electron microscopy, X-ray diffraction, Zeta potential, Fourier transform infrared, inductively coupled plasma mass spectroscopy. The formed GKPtNP are well dispersed, homogeneous with a size of 2-4 ± 0.50 nm, having a negative zeta potential (-46.1 mV) indicating good stability. 5-Fluorouracil (5FU) was loaded onto the synthesised GKPtNP, which leads to the development of a new combination of nanomedicine (5FU-GKPtNP). The in vitro drug release studies of 5FU-GKPtNP in pH 7.4 showed a sustained release profile over a period of 120 min. Agrobacterium tumefaciens induced in vitro potato tumour bioassay was employed for screening the anti-tumour potentials of GKPtNP, 5FU, and 5FU-GKPtNP. The experimental results suggested a complete tumour inhibition by 5FU-GKPtNP at a lower concentration than the GKPtNP and 5FU. Furthermore, the mechanism of anti-tumour activity was assessed by their interactions with DNA using agarose gel electrophoresis and UV-spectroscopic analysis. The electrophoresis results revealed that the 5FU-GKPtNP totally diminishes DNA and the UV-spectroscopic analysis showed a hyperchromic effect with red shift indicating intercalation type of binding with DNA. Over all, the present study revealed that the combined exposure of the nanoformulation resulted in the enhanced anti-tumour effect.

Biogenic synthesis of biopolymer-based Ag-Au bimetallic nanoparticle constructs and their anti-proliferative assessment.

This study reports an eco-friendly-based method for the preparation of biopolymer Ag-Au nanoparticles (NPs) by using gum kondagogu (GK; Cochlospermum gossypium), as both reducing and protecting agent. The formation of GK-(Ag-Au) NPs was confirmed by UV-absorption, fourier transformed infrared (FTIR), atomic force microscopy (AFM), scanning electron microscope (SEM) and transmission electron microscope (TEM). The GK-(Ag-Au) NPs were of 1-12 nm in size. The anti-proliferative activity of nanoparticle constructs was assessed by MTT assay, confocal microscopy, flow cytometry and quantitative real-time polymerase chain reaction (PCR) techniques. Expression studies revealed up-regulation of p53, caspase-3, caspase-9, peroxisome proliferator-activated receptors (PPAR) PPARa and PPARb, genes and down-regulation of Bcl-2 and Bcl-x(K) genes, in B16F10 cells treated with GK-(Ag-Au) NPs confirming the anti-proliferative properties of the nanoparticles.

Spectroscopic and biochemical correlations during the course of human lens aging.

BACKGROUND: With age, the human lens accumulates variety of substances that absorbs and fluorescence, which explains the color of yellow, brunescent and nigrescent cataract in terms of aging. The aim of this study was to assess lens fluorophores with properties comparable to those of advanced glycated end products (AGEs) in relation to age in human lenses. These fluorescent compounds are believed to be involved in the development of cataract. METHODS: Spectroscopic (UV-Vis-NIR) and fluorescence photography (CCD-Digital based image analysis) studies were carried out in randomly selected intact human lenses (2-85 years). AGE-like fluorophores were also measured in water soluble and insoluble (alkali soluble) fractions of human lenses (20-80 years). RESULTS: Our experimental findings suggest that there was a progressive shift in the absorbance characteristic of intact lens in the range of lambda210 nm-lambda470 nm. A relative increase in the absorptivity at lambda(511-520 nm), with age, was also observed. In addition, the ratio of absorptivity at lambda(511-520 nm) versus the maximum absorbance recorded at blue-end cut-off (210-470 nm) was also found to increase, with age. The fluorescent intensity in the intact lens at both UV-B (lambdaEx312 nm) and UV-A (lambdaEx365 nm) were found to be positively correlated (r2 = 0.91 & 0.94, respectively; Confidence interval 95%) upto 50 years of age. In addition, a concomitant changes in AGE- like fluorophores were also observed in the processed lens samples (soluble and insoluble fractions) along the age. A significant increase in the concentration of AGE- like fluorophores, both in intact and processed lens was observed during the period of 40 - 50 years. CONCLUSION: Based on the present investigation, it was concluded that significant changes do occur in the AGE-like fluorophores of human lenses during the period of 40-50 years.

Biogenic green synthesis of monodispersed gum kondagogu (Cochlospermum gossypium) iron nanocomposite material and its application in germination and growth of mung bean (Vigna radiata) as a plant model.

An eco-friendly green and one-pot synthesis of highly monodispersed iron (Fe) nanoparticles (NPs) by using a natural biopolymer, gum kondagogu (GK) as reducing and capping agent is proposed. The NPs synthesised were characterised by ultra-violet-visible spectroscopy, transmission electron microscopy, scanning electron microscopy and X-ray diffraction. As the concentration of gum and time increases, the intensity of NPs formation increased. The NPs were highly monodispersed with uniform circular shapes of 2-6 nm in size. The formed NPs were crystalline in nature which was confirmed by diffraction analysis. The conversion ratio of Fe ionic form to NPs was 21% which was quantified by inductively coupled plasma mass spectroscopy (ICP-MS). Fe is essential for plant growth and development. A study was conducted to examine the effect of these NPs on the growth of mung bean (Vigna radiata). The radical length and biomass was increased in seeds exposed to Fe NPs than the ions. The uptake of Fe NPs by the sprouts was also quantified by ICP-MS, in which Fe was more in mung bean seeds exposed to NPs. The α-amylase activity was increased in the seeds exposed to NPs. The observed increase in the biomass by Fe NPs and seed germination may facilitate its application in the agriculture as an important cost-effective method for plant growth.

Facile synthesis of palladium nanocatalyst using gum kondagogu (Cochlospermum gossypium): a natural biopolymer.

Palladium nanoparticles (Pd NPs) were synthesised by using gum kondagogu (GK), a non-toxic ecofriendly biopolymer. GK acted as both reducing and stabilising agent for the synthesis of Pd NPs. Various reaction parameters, such as concentration of gum, Pd chloride and reaction pH were standardised for the stable synthesis of GK reduced stabilised Pd NPs (GK-Pd NPs). The nanoparticles have been characterised using ultraviolet-visible spectroscopy, transmission electron microscopy and X-ray diffraction. Physical characterisation revealed that the gum synthesised Pd NPs were in the size range of 6.5 ± 2.3 nm and crystallised in face centred cubic (FCC) symmetry. Fourier transform infrared spectroscopy implicated the role of carboxyl, amine and hydroxyl groups in the synthesis. The synthesised Pd NPs were found to be highly stable in nature. The synthesised nanoparticles were found to function as an effective green catalyst (k = 0.182 min⁻¹) in the reduction of 4-nitrophenol by sodium borohydride, which was evident from the colour change of bright yellow (nitrophenolate; λ(max) - 400 nm) to colourless (4-AP; λ(max) - 294 nm) solution. The overall objectives of the current communication were: (i) to synthesize the Pd NPs using a green reducing/capping agent; GK and (ii) to determine the catalytic performance of the synthesised Pd NPs.

Comparative Analysis of Urinary Total Proteins by Bicinchoninic Acid and Pyrogallol Red Molybdate Methods.

BACKGROUND: The concentration of total proteins in urine is a good index of renal function, but its determination is found to be unreliable. The pyrogallol red molybdate (PRM) method for urine total proteins is being widely used in most of the hospitals because of its high sensitivity, better precision and its practicability. Bicinchoninic acid method (BCA) is also used for protein estimation and there have been no studies comparing this method with the PRM method in human urine samples. BCA method overestimates the urinary protein concentration in the presence of interfering substances. After removing the interfering substances present in the human urine samples the results of BCA method were compared with the PRM method. AIM: The purpose of the study is to identify whether the results of urine total proteins by BCA method are comparable to PRM method and can be used as an alternative to the PRM method. SETTING AND DESIGN: This is a cross-sectional study done on fresh urine specimens from the hospital laboratory, covering a wide range of protein concentrations. MATERIAL AND METHODS: Fresh urine specimens covering a wide range of protein concentrations (urine dipstick: nil, trace, 1+, 2+ and ≥ 3+) of 36 patients were analysed by both the methods. STATISTICAL ANALYSIS: Imprecision was determined by repeated analysis study and Inaccuracy was assessed by comparing the results of the patient's urine samples by both the methods using correlation plots, Bland and Altman, and Passing and Bablok regression analyses. RESULTS: The coefficient of variation and mean (SD) for the BCA method were 4.6% and 799.1 (882.5) mg/L and for the PRM method were 5.1% and 802.1 (911.9) mg/L. The Pearson correlation coefficient, r was 0.93 (p < 0.0001). Method agreement studies showed no significant constant and proportional bias between both the methods. CONCLUSION: In urine which is subjected to removal of interfering substances, the BCA results are comparable to PRM method.

Size-controlled green synthesis of silver nanoparticles mediated by gum ghatti (Anogeissus latifolia) and its biological activity.

BACKGROUND: Gum ghatti is a proteinaceous edible, exudate tree gum of India and is also used in traditional medicine. A facile and ecofriendly green method has been developed for the synthesis of silver nanoparticles from silver nitrate using gum ghatti (Anogeissus latifolia) as a reducing and stabilizing agent. The influence of concentration of gum and reaction time on the synthesis of nanoparticles was studied. UV-visible spectroscopy, transmission electron microscopy and X-ray diffraction analytical techniques were used to characterize the synthesized nanoparticles. RESULTS: By optimizing the reaction conditions, we could achieve nearly monodispersed and size controlled spherical nanoparticles of around 5.7 ± 0.2 nm. A possible mechanism involved in the reduction and stabilization of nanoparticles has been investigated using Fourier transform infrared spectroscopy and Raman spectroscopy. CONCLUSIONS: The synthesized silver nanoparticles had significant antibacterial action on both the Gram classes of bacteria. As the silver nanoparticles are encapsulated with functional group rich gum, they can be easily integrated for various biological applications.