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Abstract Exopolysaccharides (EPSs), a constitutive part of bacterial biofilm, act as a protecting sheath to the extremophilic bacteria and are of high industrial value. In this study, we elucidate a new EPS produced by thermotolerant (growth from 34–44 °C) strain Pseudomonas alcaligenes Med1 from Medano hot spring (39.1 °C surface temperature, pH 7.1) located in the Central Andean Mountains of Chile. Bacterial growth was screened for temperature tolerance (10–60 °C) to confirm the thermotolerance behaviour. Physicochemical properties of the EPS were characterized by different techniques: Scanning Electron Microscopy- Energy Dispersive X-ray Spectroscopy (SEM-EDS), Atomic Force Microscopy (AFM), High-Performance Liquid Chromatography (HPLC), Gel permeation chromatography (GPC), Fourier Transform Infrared Spectroscopy (FTIR), Nuclear Magnetic Resonance (NMR), and Thermogravimetric analysis (TGA). Whole genome of P. alcaligenes Med1 has also been studied in detail to correlate the structural and functional characteristics with genomic insight. The EPS demonstrated amorphous surface roughness composed of evenly distributed macromolecular lumps composed of mainly carbon and oxygen. The monosaccharide analysis has shown the presence of glucose, galactose, and mannose sugars at different ratios. TGA revealed the high thermal stability (315.3 °C) of the polysaccharide. The GPC has shown that Med1 is a low molecular weight polysaccharide (34.8 kDa) with low PI. The 2D-NMR linkage analysis suggests a diverse array of glycosidic bonds within the exopolysaccharide structure. The functional properties of the EPS were evaluated for food industry applications, specifically for antioxidant (DPPH, FRAP an H2O2). Extracted Med1 EPS revealed significant emulsification activity against different food grade vegetative oils (Coconut oil, Corn oil, Canola oil, Avocado oil, Sunflower oil, Olive oil, and Sesame oil). The highest 33.9% flocculation activity was observed with 60 mg L−1 EPS concentration. It showed water-holding (WHC) of 107.6% and oil-holding (OHC) capacity of 110.8%. The functional EPS produced by Pseudomonas alcaligenes Med1 from Central Andean Chilean hot spring of central Chile can be a useful additive for the food-processing industry.

Background: At present, research on facile, green synthesis of nanoparticles has significantly increased because of its fast, one-step, cost-effective, time-efficient, and non-toxic nature. In this study, we have reported a single-step green synthesis of copper nanoparticles using cell wall polysaccharides of a hot spring origin, thermotolerant Bacillus species. Result: Copper nanoparticles were characterized using UV-visible spectrophotometry, fluorescence and Fourier transform infrared spectroscopy, scanning electron microscopy with energy dispersive spectroscopy, particle size, and zeta potential analyses. UV-visible spectra of synthesized copper nanoparticles exhibited a band cantered between 220–235 nm, characteristic spectra of copper oxide nanoparticles. Infrared spectra showed the band at 490-530 cm−1 corresponding to metal-oxygen or copper nanoparticle vibration, supporting the presence of copper oxide nanoparticles in the monoclinic phase. The energy dispersive spectra of copper nanoparticles exhibited a strong signal from elemental copper. The dynamic Light Scattering pattern confirmed the nanoparticle nature of the studied sample. These nanoparticles showed preferential activity against gram-negative pathogens, Salmonella typhi and Escherichia coli. The thermodynamic nature of the nanoparticles is also established for its antibacterial actions. Conclusions: The antibacterial action and its thermodynamics reinforce the possible use of copper nanoparticles as an alternative to commercially available antimicrobials. This study may open a new path for future studies to treat harmful microorganisms resistant to traditional antibiotics in a greener way.How to cite: Banerjee A, Roy RK, Sarkar S, et al. Synthesis of hot spring origin bacterial cell wall polysaccharide-based copper nanoparticles with antibacterial property. Electron J Biotechnol 2024;68. https://doi.org/10.1016/j.ejbt.2023.11.005.

Abstract A type of high molecular weight bioactive polymers called exopolysaccharides (EPS) are produced by thermophiles, the extremophilic microbes that thrive in acidic environmental conditions of hot springs with excessively warm temperatures. Over time, EPS became important as natural biotechnological additives because of their noncytotoxic, emulsifying, antioxidant, or immunostimulant activities. In this article, we unravelled a new EPS produced by Staphylococcus sp. BSP3 from an acidic (pH 6.03) San Pedro hot spring (38.1 °C) located in the central Andean mountains in Chile. Several physicochemical techniques were performed to characterize the EPS structure including Scanning electron microscopy–energy dispersive X-ray spectroscopy (SEM–EDS), Atomic Force Microscopy (AFM), High-Performance Liquid Chromatography (HPLC), Gel permeation chromatography (GPC), Fourier Transform Infrared Spectroscopy (FTIR), 1D Nuclear Magnetic Resonance (NMR), and Thermogravimetric analysis (TGA). It was confirmed that the amorphous surface of the BSP3 EPS, composed of rough pillar-like nanostructures, is evenly distributed. The main EPS monosaccharide constituents were mannose (72%), glucose (24%) and galactose (4%). Also, it is a medium molecular weight (43.7 kDa) heteropolysaccharide. NMR spectroscopy demonstrated the presence of a [→ 6)-⍺-d-Manp-(1 → 6)-⍺-d-Manp-(1 →] backbone 2-O substituted with 1-⍺-d-Manp. A high thermal stability of EPS (287 °C) was confirmed by TGA analysis. Emulsification, antioxidant, flocculation, water-holding (WHC), and oil-holding (OHC) capacities are also studied for biotechnological industry applications. The results demonstrated that BSP3 EPS could be used as a biodegradable material for different purposes, like flocculation and natural additives in product formulation. Graphical Abstract

In the present study, a thermophilic strain designated CamBx3 was isolated from the Campanario hot spring, Chile. Based on 16S rRNA gene sequence, phylogenomic, and average nucleotide identity analysis the strain CamBx3 was identified as Bacillus paralicheniformis. Genome analysis of B. paralicheniformis CamBx3 revealed the presence of genes related to heat tolerance, exopolysaccharides (EPS), dissimilatory nitrate reduction, and assimilatory sulfate reduction. The pangenome analysis of strain CamBx3 with eight Bacillus spp. resulted in 26,562 gene clusters, 7,002 shell genes, and 19,484 cloud genes. The EPS produced by B. paralicheniformis CamBx3 was extracted, partially purified, and evaluated for its functional activities. B. paralicheniformis CamBx3 EPS with concentration 5 mg mL−1 showed an optimum 92 mM ferrous equivalent FRAP activity, while the same concentration showed a maximum 91% of Fe2+ chelating activity. B. paralicheniformis CamBx3 EPS (0.2 mg mL−1) demonstrated β-glucosidase inhibition. The EPS formed a viscoelastic gel at 45°C with a maximum instantaneous viscosity of 315 Pa.s at acidic pH 5. The present study suggests that B. paralicheniformis CamBx3 could be a valuable resource for biopolymers and bioactive molecules for industrial applications.

Xylose Isomerase (XI) is an intramolecular oxidoreductase enzyme and catalyzes the reversible conversion of ketoses and aldoses in addition to the bioconversion of ethanol from xylose in the production of bioethanol from hemicellulose. It has a broad range of industrial applications in the food and pharmaceutical sectors, particularly in the production of the sweetener high fructose corn syrup (HFCS). It is one of the most widely used industrial enzymes after protease. Taking this into consideration, four bacterial XI sources were selected based on growth temperature, i.e., psychrophile, mesophile, thermophile, and hyperthermophile, for analyzing Xylose Isomerase’s structure-function characteristics. It was found that thermophilic XI was structurally less stable than mesophilic and hyperthermophilic XI, whereas structural plasticity ran opposite towards mesophiles. The interaction of xylose isomerase (XI) with two ligands, namely Amino-2-Hydroxymethyl-Propane-1,3-Diol and (4R)-2-Methylpentane-2,4- Diol, was also studied. Mesophilic XI demonstrated better binding affinity with structurally stabilizing amino acids (Ala, Asp, Gly, Leu, and Arg). In comparison, Thermophilic XI showed nearly similar binding affinity with both Amino-2-Hydroxymethyl-Propane-1,3-Diol and (4R)-2-Methylpentane-2,4-Diol. The results of this investigation suggest that thermophilic XI, followed by mesophilic XI, would be the most appropriate for establishing process stability and sustainability in the food industry.

Rational cropping maintains high soil fertility and a healthy ecosystem. Soil microorganism is the controller of soil fertility. Meanwhile, soil microbial communities also respond to different cropping patterns. The mechanisms by which biotic and abiotic factors were affected by different cropping sequences remain unclear in the major grain-producing regions of northeastern China. To evaluate the effects of different cropping sequences under conventional fertilization practices on soil properties, microbial communities, and crop yield, six types of plant cropping systems were performed, including soybean monoculture, wheat-soybean rotation, wheat-maize-soybean rotation, soybean-maize-maize rotation, maize-soybean-soybean rotation and maize monoculture. Our results showed that compared with the single cropping system, soybean and maize crop rotation in different combinations or sequences can increase soil total organic carbon and nutrients, and promote soybean and maize yield, especially using soybean-maize-maize and maize-soybean-soybean planting system. The 16S rRNA and internal transcribed spacer (ITS) amplicon sequencing showed that different cropping systems had different effects on bacterial and fungal communities. The bacterial and fungal communities of soybean monoculture were less diverse when compared to the other crop rotation planting system. Among the different cropping sequences, the number of observed bacterial species was greater in soybean-maize-maize planting setup and fungal species in maize-soybean-soybean planting setup. Some dominant and functional bacterial and fungal taxa in the rotation soils were observed. Network-based analysis suggests that bacterial phyla Acidobacteria and Actinobacteria while fungal phylum Ascomycota showed a positive correlation with other microbial communities. The phylogenetic investigation of communities by reconstruction of unobserved states (PICRUSt) result showed the presence of various metabolic pathways. Besides, the soybean-maize-maize significantly increased the proportion of some beneficial microorganisms in the soil and reduced the soil-borne animal and plant pathogens. These results warrant further investigation into the mechanisms driving responses of beneficial microbial communities and their capacity on improving soil fertility during legume cropping. The present study extends our understanding of how different crop rotations effect soil parameters, microbial diversity, and metabolic functions, and reveals the importance of crop rotation sequences. These findings could be used to guide decision-making from the microbial perspective for annual crop planting and soil management approaches.

Salinity abiotic stress is increasing day by day due to continuous global warming and climate change. This is also becoming one of the major causes behind the reduction in crop production. Plant–bacteria interaction plays an essential role in improving crop yield without using any chemical fertilizers. The present study aims to characterize the interaction between plant-growth-promoting bacteria (PGPB) and their role in mitigating salinity stress for local variety crops. Therefore, in this work, two PGPB, namely, Bacillus proteolyticus Cyn1 and Bacillus safensis Cyn2, were isolated from rhizospheric soil of the Chilean common bean ecotype “Sapito” (Phaseolus vulgaris L.), and their PGPB traits were analyzed. Cyn1 can produce NH3 and HCN and also secrete siderophores, whereas Cyn2 produced NH3 and siderophores but responded negatively to HCN production. Both the isolated bacteria have shown a positive result for ACC deaminase production, phosphate solubilization, and catalase enzyme secretion. Under all three tested abiotic stresses, i.e., temperature, water, and salinity, both the bacteria and their consortium have demonstrated positive responses. Cyn1 under temperature stress and water stress can produce a biofilm network to combat the stress. While under salinity stress, both the PGPB isolates indicated the production of stress components and cytoplasmic inclusion bodies. Based on the response, among all other abiotic stresses, salinity stress was chosen for further plant–bacteria interaction study and growth. Visible root colonization of the bacteria has been observed in comparison to the control. The germination index was 100% for all experimental setups of seed bacterization, both under control conditions and salinity stress. Both bacteria responded with good PGP traits that helped in the growth of healthy plants after the bacterial treatment in final pot experiments. Additionally, the consortium and the plants treated with Cyn1 have demonstrated high production of photosynthetic pigments in both experimental setups. Both B. proteolyticus Cyn1 and B. safensis Cyn2 have shown promising PGP characters and efficient response against toxicity related to salinity. Hence, both of these bacteria and consortium can be used for improved agricultural production of Chilean native common beans in the near future.

Plastic waste is the biggest global problem in present times due to its constant bioaccumulation in the environment. During the last year, 367 Mt of plastics were produced in the world, of which 28.6 Mt correspond to polyurethane waste. Polyurethanes can be found in products such as adhesives, preservatives, and foams, and are often difficult to recycle. The fragmentation of plastic waste in the environment generates microplastics causing a long-term effect on our ecosystem. In search of its solution via bioremediation using enzymes, in our present study cutinase enzyme has been chosen, as it appears to be a novel candidate due to the wide variety of substrates it hydrolyzes and its presence in different microorganisms. According to physiochemical characteristics, it was found that microbial cutinase enzymes are majorly made up of aliphatic amino acids. A higher aliphatic index (more than 80) indicates the great thermostability of the enzyme. Moreover, the enzyme is found to be hydrophilic and structurally stable. The negative GRAVY value of the enzyme confirmed its stable interaction with water. It was also observed that all chosen protein models had an average of 91.10 % amino acids in the favorable regions of the Ramachandran plot. The studied microbial cutinase enzymes from both fungi Humicola insolens and actinobacterium Thermobifida fusca successfully coupled with the polyurethane resin monomers. The main interactions were found in the catalytic triad with bonds close to the urethane bonds of the ligand, in addition to having an average binding energy of − 6 kJ/mol. The interaction between the cutinases with the PUR resin as a ligand was found to be evident from our study with stable binding energies, which makes microbial cutinases potential enzymes for polyurethane waste bioremediation processes.

Background: Arsenic contamination in the ground water of rural India is a recurrent problem and decontamination is mostly based on the chemical or physical treatments until now. Microbial bioremediation is eco-friendly, cheap, time-efficient and does not produce any toxic by-products. Result: In the present study, a high arsenic tolerant bacteria Brevundimonas aurantiaca PFAB1 was isolated from Panifala hot spring located in West Bengal, India. Previously Panifala was also reported to be an arsenic-rich hot spring. B. aurantiaca PFAB1 exhibited both positive arsenic reductase and arsenite oxidase activity. It was tolerant to arsenite up to 90 mM and arsenate up to 310 mM. Electron microscopy has proved significant changes in cellular micromorphology and stalk appearance under the presence of arsenic in growth medium. Bioaccumulation of arsenic in As (III) treated cells were 0.01% of the total cell weight, while 0.43% in case of As (V) treatment. Conclusions: All experimental lines of evidence prove the uptake/accumulation of arsenic within the bacterial cell. All these features will help in the exploitation of B. aurantiaca PFAB1 as a potent biological weapon to fight arsenic toxicity in the near future.How to cite: Banerjee A, Sarkar S, Gorai S, et al. High arsenic tolerance in Brevundimonas aurantiaca PFAB1 from an arsenic-rich Indian hot spring. Electron J Biotechnol 2021:52. https://doi.org/10.1016/j.ejbt.2021.05.006

The ESBL and ACBL producing genes are responsible for bacterial resistance to a number of antibiotics. This study was undertaken to detect the distribution pattern of ESBL and ACBL producing genes among healthy Deshi and Khaki Campbell ducks that have no apparent history of antibiotic intake. For this purpose, 110 and 92 cloacal swab samples were collected from Deshi and Khaki Campbell ducks respectively. Initially samples were screened for detection of Escherichia coli, Salmonella spp. and Klebsiella pneumoniae on the basis of their cultural, biochemical and morphological properties followed by PCR confirmation. Distribution pattern of three important ESBL producing genes viz. blaCTX-M, blaTEM, blaSHV and one ACBL producing gene blaAmpC were studied thoroughly among two different segments of ducks. Significantly higher occurrence of antibiotic resistance genes was found in free range Deshi ducks compared to that of Khaki Campbell ducks. This study indicates that promotion of rearing Khaki Campbell ducks among farmers is not only beneficial for higher production but also facilitates better management of the problem associated with antibiotic resistance.

Abstract Focal cortical dysplasia (FCD) is a malformation of the cerebral cortex with poorly-defined epileptogenic zones (EZs), and poor surgical outcome in FCD is associated with inaccurate localization of the EZ. Hence, identifying novel epileptogenic markers to aid in the localization of EZ in patients with FCD is very much needed. High-throughput gene expression studies of FCD samples have the potential to uncover molecular changes underlying the epileptogenic process and identify novel markers for delineating the EZ. For this purpose, we, for the first time performed RNA sequencing of surgically resected paired tissue samples obtained from electrocorticographically graded high (MAX) and low spiking (MIN) regions of FCD type II patients and autopsy controls. We identified significant changes in the MAX samples of the FCD type II patients when compared to non-epileptic controls, but not in the case of MIN samples. We found significant enrichment for myelination, oligodendrocyte development and differentiation, neuronal and axon ensheathment, phospholipid metabolism, cell adhesion and cytoskeleton, semaphorins, and ion channels in the MAX region. Through the integration of both MAX vs non-epileptic control and MAX vs MIN RNA sequencing (RNA Seq) data, PLP1, PLLP, UGT8, KLK6, SOX10, MOG, MAG, MOBP, ANLN, ERMN, SPP1, CLDN11, TNC, GPR37, SLC12A2, ABCA2, ABCA8, ASPA, P2RX7, CERS2, MAP4K4, TF, CTGF, Semaphorins, Opalin, FGFs, CALB2, and TNC were identified as potential key regulators of multiple pathways related to FCD type II pathology. We have identified novel epileptogenic marker elements that may contribute to epileptogenicity in patients with FCD and could be possible markers for the localization of EZ.

The ESBL producing genes are responsible for bacterial resistance to number of antibiotics whereas Shiga toxin producing genes are responsible for bacterial virulence. The association between ESBL producing genes and Shiga toxin producing E. coli (STEC) may pose bigger threat in the battle of antibiotic resistance. This study was conducted to determine the prevalence of Shiga-toxin-producing Escherichia coli (STEC) in ducks reared in organized and unorganized sectors from different agro climatic zones of West Bengal, India and to study their relationship with extended spectrum beta-lactamase (ESBL) production. Total 202 cloacal swab samples were collected from both indigenous ducks reared in backyards sector (110 samples) and Khaki Campbell Ducks reared in organized farm (92 samples). Initially the samples were screened for detection of E. coli on the basis of their cultural, morphological and biochemical properties followed by PCR confirmation for E. coli 16S rRNA. E. coli isolates were subjected to multiplex PCR to detect the presence of shiga toxin producing genes such as stx1, stx2, eaeA and ehxA. STEC isolates were screened phenotypically for production of ESBL and ACBL by double disk diffusion method and subsequently PCR detection for blaCTX-M, blaTEM, blaSHV and blaAmpC genes were performed. Serotyping of all the STEC was also done. Out of 202 samples total 109 were confirmed to be E. coli positive. Out of them total 27 (24.77 %) E. coli isolates were detected to be positive for STEC. Higher prevalence of STEC was observed in unorganized sector compared to the organized sector. Positive association (P < 0.05) was observed between STEC and ESBL production. This study indicates that the duck may play an important role in transmission of Siga-toxin-producing E. coli (STEC) as well as antibiotic resistance genes to human beings, other birds, animals and environment also.

Glutamate-receptor-mediated hyperexcitability contributes to seizure generation in temporal lobe epilepsy (TLE). Tryptophan–kynurenine pathway (TKP) metabolites regulate glutamate receptor activity under physiological conditions. This study was designed to investigate alterations in the levels of TKP metabolites and the differential regulation of glutamatergic activity by TKP metabolites in the hippocampus, anterior temporal lobe (ATL), and neocortex samples of a lithium–pilocarpine rat model of TLE. We observed that levels of tryptophan were reduced in the hippocampus and ATL samples but unaltered in the neocortex samples. The levels of kynurenic acid were reduced in the hippocampus samples and unaltered in the ATL and neocortex samples of the TLE rats. The levels of kynurenine were unaltered in all three regions of the TLE rats. The magnitude of reduction in these metabolites in all regions was unaltered in the TLE rats. The frequency and amplitude of spontaneous excitatory postsynaptic currents were enhanced in hippocampus ATL samples but not in the neocortex samples of the TLE rats. The exogenous application of kynurenic acid inhibited glutamatergic activity in the slice preparations of all these regions in both the control and the TLE rats. However, the magnitude of reduction in the frequency of kynurenic acid was higher in the hippocampus (18.44 ± 2.6% in control vs. 30.02 ± 1.5 in TLE rats) and ATL (16.31 ± 0.91% in control vs. 29.82 ± 3.08% in TLE rats) samples of the TLE rats. These findings suggest the differential regulation of glutamatergic activity by TKP metabolites in the hippocampus, ATL, and neocortex of TLE rats.

Various microorganisms thrive under extreme environments, like hot springs, hydrothermal vents, deep marine ecosystems, hyperacid lakes, acid mine drainage, high UV exposure, and more. To survive against the deleterious effect of these extreme circumstances, they form a network of biofilm where exopolysaccharides (EPSs) comprise a substantial part. The EPSs are often polyanionic due to different functional groups in their structural backbone, including uronic acids, sulfated units, and phosphate groups. Altogether, these chemical groups provide EPSs with a negative charge allowing them to (a) act as ligands toward dissolved cations as well as trace, and toxic metals; (b) be tolerant to the presence of salts, surfactants, and alpha-hydroxyl acids; and (c) interface the solubilization of hydrocarbons. Owing to their unique structural and functional characteristics, EPSs are anticipated to be utilized industrially to remediation of metals, crude oil, and hydrocarbons from contaminated wastewaters, mines, and oil spills. The biotechnological advantages of extremophilic EPSs are more diverse than traditional biopolymers. The present review aims at discussing the mechanisms and strategies for using EPSs from extremophiles in industries and environment bioremediation. Additionally, the potential of EPSs as fascinating biomaterials to mediate biogenic nanoparticles synthesis and treat multicomponent water contaminants is discussed.

Background: Textile industry not only plays a vital role in our daily life but also a prominent factor in improving global economy. One of the environmental concern is it releases huge quantities of toxic dyes in the water leading to severe environmental pollution. Bacterial laccase and azoreductase successfully oxidize complex chemical structure of nitrogen group-containing azo dyes. Additionally, the presence of textile dye infuriates bacterial peroxidase to act as a dye degrading enzyme. Our present study deals with three textile dye degrading enzymes laccase, azoreductase, and peroxidase through analyzing their structural and functional properties using standard computational tools. Result: According to the comparative analysis of physicochemical characteristics, it was clear that laccase was mostly made up of basic amino acids whereas azoreductase and peroxidase both comprised of acidic amino acids. Higher aliphatic index ascertained the thermostability of all these three enzymes. Negative GRAVY value of the enzymes confirmed better water interaction of the enzymes. Instability index depicted that compared to laccase and preoxidase, azoreductase was more stable in nature. It was also observed that the three model proteins had more than 90% of total amino acids in the favored region of Ramachandran plot. Functional analysis revealed laccase as multicopper oxidase type enzyme and azoreductase as FMN dependent enzyme, while peroxidase consisted of α-β barrel with additional haem group. Conclusion: Present study aims to provide knowledge on industrial dye degrading enzymes, choosing the suitable enzyme for industrial set up and to help in understanding the experimental laboratory requirements as well.How to cite: Sarkar S, Banerjee A, Chakraborty N, et al. Structural-functional analyses of textile dye degrading azoreductase, laccase and peroxidase: a comparative in silico study. Electron J Biotechnol 2020;43. https://doi.org/10.1016/j.ejbt.2019.12.004 Keywords: Acidic amino acids, Azo dyes, Azoreductase, Basic amino acids, Dye degradation, Laccase, Peroxidase, Textile industry, Thermostable three enzymes

Marine microalgae has been attracting the researcher’s attention for centuries. Development on microalgal research is majorly favoured by its medicinal, pharmaceutical or cosmeceutical properties. The advancement in the investigation related to microalgal products have been concentrated in the coastal zones because of the greater supply of raw material. The Southern Ocean, highly productive and relatively poorly studied ecosystem, constitutes approximately 10% of the global volume of the oceans. In this study, marine microalgae Chlorella sp. PR-1 was isolated from the Southern Ocean (Indian Sector) for the identification of bioactive antibacterial compounds. The antimicrobial activity of this microalgal extracts was evaluated against three gram positive (Staphylococcus aureus, Bacillus licheniformis, and Bacillus subtilis) and three gram negative bacteria (Pseudomonas aeruginosa, Salmonella typhimurium, and Escherichia coli). The extract showing antibacterial activity was further purified by thin layer and column chromatography. The antimicrobial activity was again evaluated and confirmed with the purified fraction against the same bacteria. The identification of the functional groups in the purified fraction was performed by infrared spectroscopic analysis. Based on gas chromatographic and mass spectroscopic analysis, the principle bioactive compound was proved to be 2,4-bis (1,1-dimethylethyl)- phenol. Thus, the bioactive compound isolated from marine microalga of Southern Ocean origin may be a novel alternative source of antibacterials in the future.

The peritumoral regions of WHO grade II gliomas, like astrocytoma and oligodendroglioma, have been reported to show epileptiform activities. An imbalance of glutamatergic and GABAergic mechanisms is primarily responsible for the generation of epileptiform activities. Here we have compared the electrophysiological properties of pyramidal neurons in intraoperative peritumoral specimens obtained from glioma patients with (GS) and without (GN) a history of seizures at presentation. Histology and immunohistochemistry were performed to assess the infiltration of proliferating cells at the peritumoral tissues. Whole-cell patch clamp technique was performed to measure the spontaneous glutamatergic and GABAergic activity onto pyramidal neurons in the peritumoral samples of GS (n = 11) and GN (n = 15) patients. The cytoarchitecture of the peritumoral tissues was devoid of Ki67 immuno-positive cells. We observed a higher frequency of spontaneous glutamatergic and GABAergic activities onto pyramidal neurons of the peritumoral samples of GS patients. Our findings suggest that, in spite of similar histopathological features, the pyramidal neurons in the peritumoral samples of GS and GN patients showed differences in spontaneous excitatory and inhibitory synaptic neurotransmission. An alteration in postsynaptic currents may contribute to the spontaneous epileptiform activity in GS patients.

Extremophilic microorganisms often produce novel bioactive compounds to survive under harsh environmental conditions. Exopolysaccharides (EPSs), a constitutive part of bacterial biofilm, are functional biopolymers that act as a protecting sheath to the extremophilic bacteria and are of high industrial value. In this study, we elucidate a new EPS produced by thermophilic Bacillus haynesii CamB6 from a slightly acidic (pH 5.82) Campanario hot spring (56.4 °C) located in the Central Andean Mountains of Chile. Physicochemical properties of the EPS were characterized by different techniques: Scanning electron microscopy- energy dispersive X-ray spectroscopy (SEM-EDS), Atomic Force Microscopy (AFM), High-Performance Liquid Chromatography (HPLC), Gel permeation chromatography (GPC), Fourier Transform Infrared Spectroscopy (FTIR), 1D and 2D Nuclear Magnetic Resonance (NMR), and Thermogravimetric analysis (TGA). The EPS demonstrated amorphous surface roughness composed of evenly distributed macromolecular lumps. GPC and HPLC analysis showed that the EPS is a low molecular weight heteropolymer composed of mannose (66%), glucose (20%), and galactose (14%). FTIR analysis demonstrated the polysaccharide nature (–OH groups, Acetyl groups, and pyranosic ring structure) and the presence of different glycosidic linkages among sugar residues, which was further confirmed by NMR spectroscopic analyses. Moreover, D-mannose α-(1→2) and α-(1→4) linkages prevail in the CamB6 EPS structure. TGA revealed the high thermal stability (240 °C) of the polysaccharide. The functional properties of the EPS were evaluated for food industry applications, specifically as an antioxidant and for its emulsification, water-holding (WHC), oil-holding (OHC), and flocculation capacities. The results suggest that the study EPS can be a useful additive for the food-processing industry.

Enhanced spontaneous GABAA receptor activity is associated with focal cortical dysplasia (FCD), a developmental malformation of the cerebral cortex. Clinical manifestations in FCD vary with age at epilepsy onset with a more favorable prognosis in patients with late-onset (LO) compared to that in cases with early-onset (EO). This study was designed to test the hypothesis in FCD that spontaneous GABAA receptor-mediated epileptogenicity depends on the age at epilepsy onset and varies between patients with early and late-onset age in FCD. To this end, brain specimens were obtained from the maximal spiking region (MAX) and minimal spiking region (MIN) of the epileptic foci of EO (n = 14, mean age = 10.6 ± 2.9 years) and LO (n = 10, mean age = 27 ± 5.6 years) patients undergoing electrocorticography (ECoG) guided surgery. The whole-cell patch-clamp technique was used to record spontaneous GABAergic currents from normal-looking pyramidal neurons in slice preparations of resected brain samples. We detected higher frequency and amplitude of GABAergic events in MAX samples compared to MIN samples of LO patients, while they were comparable in MIN and MAX samples of EO patients. Further GABAergic activity in the MIN and MAX samples of EO patients was higher than the MIN samples of LO patients. This suggests that in LO patients, GABAA receptor-mediated epileptogenicity is confined only to the high spiking areas, but in EO patients, it affects low spiking regions as well.

Xylans are the second most abundant form of hemicelluloses and are the second most abundant polysaccharide in nature after cellulose. To degrade xylan, microbes produce mainly xylanase enzyme. Wide range of microorganisms like fungi, bacteria, yeast, marine algae etc. are capable of producing xylanase. Main source of xylanase is fungi but industrial production of bacterial xylanase is low cost, easy downstream process and high production rate. To understand primary, secondary and tertiary structure of xylanase, in silico composition of amino acids, basic physiological characteristics; viz., pI, molecular weight, instability index, GRAVY, molar extinction coefficient, secondary structure, presence of functional domain and motifs, phylogenetic tree, salt bridge compositions are determined. In silico study of xylanase focused on 36 different bacterial sources are performed by retrieving FASTA and PDB sequences using RCSB PDB. FASTA and PDB files are proceed further in ExPASy-ProtParam, RAMPAGE, QMEAN, MEME, PSIPRED, InterProScan, MOTIF scan, ERRAT, Peptide cutter, ESBRI and MEGA 7. The instability index range (16.90–38.78) clearly indicates that the protein is highly stable. α-helix mean value (27.11%) infers the protein is dominated by α-helix region. The aliphatic index (39.80–90.68) gives information that the protein is highly thermostable, prevalence by alanine amino acid in aliphatic side chain. No transmembrane domain was found in the protein which confirms the enzyme is extracellular in nature. Ancestor chart analysis confirmed that it is a part of carbohydrate metabolic process and more specifically a member of glycoside hydrolase super family. Keywords: Bacterial xylanase, Phylogenetic relationships, Physical parameters, Structural and functional analysis

This study investigated the biocomposite pectin films enriched with murta (Ugni molinae T.) seed polyphenolic extract and reinforced by chitin nanofiber. The structural, morphological, mechanical, barrier, colorimetric, and antioxidant activity of films were evaluated. The obtained data clearly demonstrated that the addition of murta seed extract and the high load of chitin nanofibers (50%) provided more cohesive and dense morphology of films and improved the mechanical resistance and water vapor barrier in comparison to the control pectin film. The antioxidant activity ranged between 71% and 86%, depending on the film formulation and concentration of chitin nanofibers. The presented results highlight the potential use of chitin nanofibers and murta seed extract in the pectin matrix to be applied in functional food coatings and packaging, as a sustainable solution.

Deep marine microorganisms survive under extreme ecological settings and harsh environmental conditions of low temperature, high salinity, and high atmospheric pressure making it significant of scientific interest. Southern Ocean (SO) is one such example of deep marine ecosystem and the microorganisms inhabiting in such hostile environment may produce different bioactive secondary metabolites. SO (Indian Sector) is relatively less documented in terms of microbial composition and community dynamics. The present study involves isolation of exopolysaccharides (EPSs) from three potent SO (Indian Sector) bacteria, optimization of the EPS production and partial characterization of them. Three different EPSs show varying structural conformation, that is from porous to strong flakes mimicking polymeric structure with C/N ratio ranging between 4 - 11. FTIR spectra have exhibited the presence of different active groups of carbohydrate moieties, water molecules and protein-associated amides. EPSs produced by marine microorganisms show high biotechnological promises such as drug carrier in pharmaceutical field, emulsifier and cryo- protectant in food-processing industry, detoxification of petrochemical oils and much more. The three bacterial isolates in this study showed potential of producing EPS biopolymer that can be further explored in terms of its proper biotechnological applications.

Aim: This study is focused on GABRG2 gene sequence variations in patients with mesial temporal lobe epilepsy (mTLE). The GABAAreceptor is a heteropentameric receptor and alpha-1 beta-2 gamma-2 subunits combination is most abundant and present in almost all regions of the brain. The gamma-2 subunit (GABRG2) gene mutations have been reported in different epilepsy pathologies. In the present study we have looked for GABRG2 gene sequence variations in patients with mTLE. Materials and Methods: Twenty patients (12 females and eight males, age 4.6-38 years) with MTLE were recruited for this investigation. Patients were recommended for epilepsy surgery after all clinical investigations as per the epilepsy protocol. Ethnically matched glioma or meningioma patients were considered as nonepileptic controls. During temporal lobectomy of amygdalohippocampectomy, hippocampal brain tissue samples were resected guided by intraoperative electrocorticography (ECoG) activity. All 11 exons of GABRG2 gene with their flanking intronic regions were amplified by polymerase chain reaction (PCR) and screened by DNA sequencing analysis for sequence variations. Statistical Analysis Used: Comparison of allele frequencies between patient and control groups was determined using a c2 test. Results and Conclusions: Total five DNA sequence variations were identified, three in exonic regions (c.643A > G, rs211035), (c.T > A, rs424740), and (c.C > T, rs418210) and two in intronic regions (c.751 + 41A > G, rs211034) and (c.751 + 52G > A, rs 34281163). Allele frequencies of variants identified in this study did not differ between patients and normal controls. Thus, we conclude that GABRG2 gene may not be playing significant role in the development of epilepsy or as a susceptibility gene in patients with MTLE in Indian population.

Municipal solid waste was collected from Varanasi’s municipal solid waste dumpsite and sorted for organic fraction present in it. Current work showed the consequences of calcium hydroxide or lime digestion on organic fraction of municipal solid waste of Varanasi, India. The organic fraction of municipal solid waste sample was digested with desired amount of calcium hydroxide. The different concentrations (0.1, 0.2, and 0.5 %) of calcium hydroxide was blended separately to substrates (10 % total solid) at 30-35 ºC in 3 different beakers denoted by A1, A2 and A3, respectively of 2 L capacity. Experiments of hydrolysis step were conducted on all three samples for evaluation of chemical oxygen demand, pH and volatile fatty acids content in sample. The parameters measured during experiments were chemical oxygen demand, biogas production, total solid, volatile solids, total Kjeldahl nitrogen and total organic carbon. Rate enhancement of anaerobic digestion and biogas production were occurred for calcium hydroxide digested samples. Therefore calcium hydroxide can be used as an effective alkali for the digestion of organic fraction of municipal solid waste.

Aim of the study: Activating Transforming factor 3 (ATF3) is a stress induced gene and closely associated with neuro-inflammation while Transforming growth Factor Beta (TGFβ) signalling is also reported to be involved in neuro-inflammation and hyper-excitability associated with drug resistant epilepsy. Animal model studies indicate the involvement of ATF3 and TGFβ receptors to promote epileptogenesis. Human studies also show that TGFβ signalling is activated in MTLE-HS. However, lack of studies on ATF3 and TGFβRI expression in MTLE-HS patients exists. We hypothesize that ATF3 and TGFβRI might be expressed in hippocampi of patients with MTLE-HS and playing role in epileptogenesis. Materials & methods: Protein expression of ATF3 and TGFβRI was performed by western blotting. Localisation of ATF3 was performed by immunohistochemistry and immunoflorescence. Results: Protein expression of ATF3 and TGFβRI was significantly up-regulated in hippocampi of patients as compared to controls. Also ATF3 IR was significantly expressed in hippocampi of patients and ATF3 was expressed predominantly in cytoplasm as compared to nucleus. No correlation was found between ATF3 expression and epilepsy duration and seizure frequency. Conclusions: ATF3 and TGFβRI are both important players in neuro-inflammation and might potentiate epileptogenesis in these patients. [ABSTRACT FROM AUTHOR]