Purification and evaluation of 2, 4-di-tert butylphenol (DTBP) as a biocontrol agent against phyto-pathogenic fungi.

A fungal strain, Marasmiellus sp (PUK64), isolated from the mangrove forests in Muthupet, Tamil Nadu, East coast of India, along with others were screened for the search of potent bioactive compounds. A phenolic compound, 2,4-di-tert-butylphenol (DTBP), was isolated from the most promising strain PUK64 and its chemical structure was ascertained. DTBP demonstrated remarkable antifungal activity against the phytopathogenic fungi Aspergillus oryzae, Curvularia lunata and Fusarium verticillioides. In an in-vitro experimental setup, DTBP suppressed the growth of all three fungi, among which F. verticillioides was found to be highly susceptible. This effect relates with the inhibition of spore germination and hyphal growth that we observed. DTBP showed high affinity with the F. verticillioides's ß-tubulin protein (determined by ligand-protein docking) as compared to the standard fungicide carbendazim (CBZ). Molecular docking and simulation studies of DTBP with target ß-tubulin further confirmed the potential of ß-tubulin binding in F. verticillioides. To our knowledge, this is the first report on DTBP-mediated biocontrol of phytopathogenic fungi, produced by Marasmiellus sp. PUK64 that can be potent inhibitor of ß-tubulin protein of F. verticillioides.

Correlation of SARS-CoV-2 Serology and Clinical Phenotype Amongst Hospitalised Children in a Tertiary Children's Hospital in India.

INTRODUCTION: Children usually present with minimal or no symptoms of COVID-19 infection. Antibody responses to SARS-CoV-2 in children from low- and middle-income countries (LMIC) have not been well described. We describe the prevalence of anti-SARS-CoV-2 antibodies and clinical phenotype of seropositive children admitted to a tertiary children's hospital in South India. METHODS: To determine the seropositivity and describe the clinical characteristics of COVID-19 infection amongst hospitalised children, we performed a prospective clinical data collection and blood sampling of children admitted to Kanchi Kamakoti CHILDS Trust Hospital, Chennai, India over 4 months of the COVID-19 pandemic. In seropositive children, we compared antibody titres between children with and without PIMS-TS. RESULTS: Of 463 children, 91 (19.6%) were seropositive. The median (range) age of seropositive children was 5 years (1 month-17 years). Clinical presentation was consistent with Paediatric inflammatory multisystem syndrome associated or related with SARS-CoV-2 infection (PIMS-TS) in 48% (44/91) of seropositive children. The median (range) antibody titre was 54.8 (11.1-170.9) AU/ml among all seropositive children. The median antibody titre among the children with PIMS-TS (60.3 AU/mL) was significantly (p = 0.01) higher when compared to the children without PIMS-TS (54.8 AU/mL). CONCLUSION: We describe the antibody responses to SARS-CoV-2 amongst hospitalised children in a LMIC tertiary children's hospital. Almost half of the seropositive children had PIMS-TS. Antibody levels may be helpful in the diagnosis and disease stratification of PIMS-TS. LAY SUMMARY: Children usually present with minimal or no symptoms of COVID-19 infection. However, Multisystem Inflammatory Syndrome in Children (MIS-C) or Paediatric inflammatory multisystem syndrome associated or related with SARS-CoV-2 infection (PIMS-TS) has emerged as a distinctive paediatric illness related to SARS-CoV-2. Recently, antibody testing for SARS-CoV-2 is being used increasingly as a diagnostic test for PIMS-TS. However, data on the antibody responses to SARS-CoV-2 in children are sparse. We, therefore, attempted to identify the seropositivity and describe the clinical spectrum of COVID-19 infection amongst infants and children getting hospitalised in a children's hospital in south India. Nearly one-fifth of the hospitalised children tested serology positive over 4 months. Antibody levels in children with PIMS-TS were significantly higher in comparison to the other two groups (acute COVID-19 infection and children without PIMS-TS). Results from our study suggest that all children are at risk of COVID-19 infection though they may present with mild illness or no symptoms. We also observed that antibody testing may have a possible role in diagnosis of PIMS-TS.

Correlation of SARS-CoV-2 serology and clinical phenotype amongst hospitalised children in a tertiary children's hospital in India

IntroductionChildren usually present with minimal or no symptoms of SARS-CoV-2 infection. Antibody responses to SARS-CoV-2 in children from low- and middle-income countries (LMIC) have not been well described. We describe the prevalence of anti-SARS-CoV-2 antibodies and clinical phenotype of seropositive children admitted to a tertiary childrens hospital in South India. MethodsTo determine the seropositivity and describe the clinical characteristics of SARS-CoV-2 infection amongst hospitalised children, we performed a prospective clinical data collection and blood sampling of children admitted to Kanchi Kamakoti CHILDS Trust Hospital, Chennai, India over 4 months of the COVID-19 pandemic. In seropositive children, we compared antibody titres between children with and without PIMS-TS. ResultsOf 463 children, 91 (19.6%) were seropositive. The median (range) age of seropositive children was 5 years (1 month - 17 years). Clinical presentation was consistent with Paediatric inflammatory multisystem syndrome temporally associated with SARS-CoV-2 infection (PIMS-TS) in 48% (44/91) of seropositive children. The median (range) antibody titre was 54.8 (11.1-170.9) AU/ml among all seropositive children. The median antibody titre among the children with PIMS-TS (60.3 AU/mL) was significantly (p=0.01) higher when compared to the children without PIM-TS (54.8 AU/mL). ConclusionWe describe the antibody responses to SARS-CoV-2 amongst hospitalised children in a LMIC tertiary childrens hospital. Almost half of the seropositive children had PIMS-TS. Antibody levels may be helpful in the diagnosis and disease stratification of PIMS-TS. Lay summaryChildren usually present with minimal or no symptoms of SARS-CoV-2 infection. However, Paediatric inflammatory multisystem syndrome temporally associated with SARS-CoV-2 infection (PIMS-TS) has emerged as a distinctive paediatric illness related to SARS-CoV-2. Recently, antibody testing for SARS-CoV-2 is being used increasingly as a diagnostic test for PIMS-TS. However, data on the antibody responses to SARS-CoV-2 in children is sparse. We therefore, attempted to identify the seropositivity and describe the clinical spectrum of SARS-CoV-2 infection amongst infants and children getting hospitalised in a childrens hospital in south India. Nearly one-fifth of the hospitalised children tested serology positive over 4 months. Antibody levels in children with PIMS-TS were significantly higher in comparison to the other two groups (acute SARS-CoV-2 infection and children without PIMS-TS). Results from our study suggest that all children are at risk of SARS-CoV-2 infection though they may present with mild illness or no symptoms. We also observed that antibody testing may have a possible role in diagnosis of PIMS-TS.

K - Shrinkage Function for ECG Signal Denoising.

ECG signals is a graphical way of recording the electrical actions of the heart for the various diagnostic purposes. ECG signals are affected by various noises such as Electrode Contact, Baseline Wandering, Motion artifact, Power-line interference, Muscle Contractions, and Electrosurgical noise during data acquisition. Denoising is a technique which is used for removing the noise in ECG signals which keeps the useful information. In this paper, a new category of Wavelet shrinkage methods is proposed. The white Gaussian noise is mixed with the ECGs for simulation and tested with the new class of shrinkage function and is compared with the other wavelet shrinkage functions such as hard and soft shrinkage. The performance measures such as Signal to Noise Ratio (SNR) and Percent Root mean - square Difference (PRD) etc. are used to examine the performance of various shrinkage functions. The experimental result shows that it gives better MSE over conventional shrinkage functions.

Receive diversity based transmission data rate optimization for improved network lifetime and delay efficiency of Wireless Body Area Networks.

Wireless Body Area Network (WBAN) has become the emerging technology due to its ability to provide intelligent and cost-effective healthcare monitoring solution. The biological sensors used in WBAN are energy-constrained and required to be functional for a longer duration. Also, the sensed data should be communicated in reasonable time. Therefore, network lifetime and delay have become the primary concerns in the design of WBAN. In this paper, Receive Diversity based Transmission Data Rate Optimization (RDTDRO) scheme is proposed to improve the network lifetime and delay efficiency of Multi level-Quadrature Amplitude Modulation (M-QAM) based WBAN. In the proposed RDTDRO scheme, minimum energy consumption is ensured by optimizing the transmission data rate with respect to a given transmission distance and number of receive antennas while satisfying the Bit Error Rate (BER) requirements. The performance of proposed RDTDRO is analyzed in terms of network lifetime and delay difference and is compared with conventional Baseline and Rate optimized schemes. The results show that at a transmission distance of 0.3 m, the proposed RDTDRO scheme with a receive diversity order of 4 achieves 1.30 times and 1.27 times improvement in network lifetime over conventional Baseline and Rate optimized schemes respectively. From the results, it is also evident that at a transmission distance of 0.3 m, the proposed RDTDRO scheme with a receive diversity order of 4 is delay efficient as it achieves delay difference of 0.75 µs and 0.29 µs over conventional Baseline and Rate optimized schemes respectively.

Identification of inhibitor against H. pylori HtrA protease using structure-based virtual screening and molecular dynamics simulations approaches.

The HtrA protease of Helicobacter pylori, which efficiently colonizes at the gastric epithelial of host cells, disrupts the mucosal integrity of E-cadherin and spreads inflammatory diseases including gastric cancer by cleaving the cell-cell adhesion of the host. The lack of knowledge on the molecular diversity, structural and functional behavior of HpHtrA necessitated the present study to explore its inhibition mechanism. At first, the similarity of HpHtrA with other gastro-intestinal pathogenic HtrA bacteria and its remote relationship with the Human HtrA homologs were ensured by the phylogenetic analysis and hence was identified as a novel therapeutic target for further design of inhibitors. The three dimensional structure of HpHtrA was modeled and simulated to achieve its stable conformation and was used as a receptor to screen for the possible lead compound through virtual screening (using ∼ 1.3 million compounds). Molecular dynamics simulations followed by the binding energy analysis revealed the affinity of the compound 300040 in forming a stable complex with HpHtrA and thereby revealed its potent role in inhibiting HpHtrA. It is also worthy to mention that, structurally, the ligand binding at the catalytic site of HpHtrA is mainly facilitated by the significant dynamics of L2 loop. Based on the present study, the hydroxyl-piperidine with 4-aminopiperidine scaffold is proposed to be one of the best possible lead compounds for the inhibition of H. pylori.

Pharmacophore modelling and electronic feature analysis of hydroxamic acid derivatives, the HIV integrase inhibitors.

Hydroxamic acid derivatives with metal ion binding properties were collected from the literature to generate a pharmacophore and 3D-QSAR model for HIV strand transfer inhibition. The derived pharmacophore model (AAAHRR) recognizes both metal ion binding site and hydrophobic group. The QSAR model generated using this hypothesis expressed statistical significance (r(2) = 0.971 for the training set and q(2) = 0.913 for the test set). The ability of this pharmacophore model to retrieve other metal ion binding inhibitors was examined by screening the ChemBank database (ligandinfo) incorporated with 10 known strand transfer inhibitors. The studied favourable and unfavourable contours of chemical features (H-bond donor, acceptor and hydrophobic sites) revealed the role of hydrophobic substitution at the fluorobenzene ring and cyclization of the metal ion binding hydroxamic acid in effective integrase inhibition. Analysis of the frontier orbitals, HOMO and LUMO revealed that the nucleophilic / electrophilic interactions depend on the significant overlapping observed at the azaindole and hydroxamic acid groups. In essence, the generated pharmacophore model is competent enough to disclose the essential site-specific interactions involved in the inhibition of HIV integrase, and hence can be used in virtual screening to identify novel scaffolds as leads with increased anti-viral potency.

Facile microwave-combustion synthesis of wurtzite CdS nanoparticles.

In this study we first report microwave-combustion synthesis of faceted CdS nanoparticles by using cadmium thiocyanate complex as a single source precursor. This is the first example of a metal-thiocyanate (M-SCN) complex being used as a source for metal sulfides (M-S) preparation in a microwave-combustion process. The synthesized CdS was characterized using X-ray diffraction (XRD), field mission scanning electron microscopy (FE-SEM), and high-resolution transmission electron microscopy (HR-TEM). The by-product assisted combustion synthesis yields CdS nanoparticles with the mixtures of octahedral geometries, hexagonal, and triangle plate morphologies and the sizes were found to be 100 nm to 5 microm. The XRD patterns imply the formation of well crystallized wurtzite CdS. The influence of cadmium and sulfur precursors and microwave irradiation time on the morphology of CdS nanoparticle was also investigated. The cadmium and sulfur precursors strongly influenced the CdS morphology and increasing the microwave irradiation time and intensity has no effect on the CdS morphology. In addition, a plausible mechanism of CdS nanoparticle formation has been proposed in this research.

Template-free synthesis of self-assembled Co3O4 micro/nanocrystals.

In this article, we have reported the fabrication of various morphological porous Co3O4 by thermal decomposition of cobalt oxalate at open atmospheric conditions. Uniform cobalt oxalate microrods and microneedles were synthesized without using any surfactants or templates in large scale. The cobalt oxalate preparation method was played a crucial role on the crystal structure and its morphology. The as prepared cobalt oxalates and its corresponding cobalt oxides were characterized by using the thermogravimetric analysis, X-ray diffraction (XRD), field-emission scanning electron microscope (FE-SEM), transmission electron microscopy (TEM) and nitrogen adsorption analysis. The XRD results indicated that the orthorhombic and monoclinic cobalt oxalates were formed in different experimental conditions. The influence of preparation method of cobalt oxalates and cobalt precursors on the final morphology has been investigated. The M-H loop of the Co3O4 porous microrods and microneedles showed the presence of paramagnetic properties at room temperatures. A plausible mechanism of both cobalt oxalates and Co3O4 formation was proposed based on the experimental results.

Template-free synthesis of beta-In2S3 superstructures and their photocatalytic activity.

In this article, we have successfully fabricated various morphological beta-Indium sulfide (In2S3) superstructures by using indium thiocyanate complex at acidic pH. All the synthesis has been performed by a template-free, hydrothermal method at 195 degrees C for 3 h. The photocatalytic activity of synthesized In2S3 have been investigated by using UV-B (lamda = 365 nm) light with Methyl Orange dye as a model pollutant. The synthesized photocatalyst was characterized by using XRD, FE-SEM, HR-TEM, DRS spectra and nitrogen adsorption analysis. The influence of indium precursors and solvents on the morphology as well as the surface properties has also been discussed. The XRD result shows that cubic phase beta-In2S3 formed under all experimental conditions. A plausible mechanism of the In2S3 microsphere formation has been discussed based on experimental observations.

Synergistic effect of hydroxypropyl-beta-cyclodextrin encapsulated soluble ferrocene and the gold nanocomposite modified glassy carbon electrode for the estimation of NO in biological systems.

An electrochemical assay for sensing NO in biological systems is described in this paper. The ferrocene mediated reduction of NO, facilitated by the gold nanocomposite modified glassy carbon electrode is followed by an amperometric procedure. The analytical protocol involves the modification of a glassy carbon electrode by an overlayer of Au nanocomposites prepared through galvanic reduction. Additional overlayers can be built on the surface by repetition of the procedure. The modification leads to the decrease of the over-potential required for the analysis and results in a non-biofouling surface. Since the procedure is based on the electrochemical reduction of NO, the potential interferences from species like dopamine, ascorbic acid, etc., are overcome. The sensitivity, detection limit and response time achieved through this protocol for the modified electrode containing three Au overlayers are 0.03 nA/nM, 25.75 nM and <5 s. Analysis of NO has been carried out in real samples like liver extract, peripheral blood mononuclear cells (PBMCs) and miconazole nitrate ointment and the values obtained are comparable with that obtained by Griess analysis.

Electrochemical assay of the nitrate and nitrite reductase activities of Rhizobium japonicum.

This work describes an electrochemical method for the determination of the nitrate and nitrite reductase activities of Rhizobium japonicum. The advantage of the method lies in the use of whole cells for the analysis and we earlier developed this protocol for the assay of NO. The results obtained are comparable to the spectrophotometric Griess assay. As the method is based on electrochemical reduction, the commonly interfering biological components like ascorbic acid, uric acid, dopamine, etc., will not interfere with the analysis. This method can be extended to the fabrication of biosensors for nitrate and nitrite using the same principle.

Detection of pathogenic leptospires in animals by PCR based on lipL21 and lipL32 genes.

Efficacy of primers capable of amplifying conserved outer membrane protein (OMP) genes i.e., lipL21 and lipL32 of Leptospira strains was tested for rapid and early diagnosis of the leptospirosis using a polymerase chain reaction (PCR). These OMP genes were found to be conserved in various leptospiral serovars viz., Canicola, Pomona, Icterohaemorrhagiae, Pyrogenes, Sejroe, Grippotyphosa, Ballum and Tarassovi as PCR products of 561 bp and 756 bp were obtained by PCR employing lipL21 and lipL32 based primers, respectively, in all these serovars. Absence of such amplicons in DNA extracted from Pasteurella, Campylobacter and Brucella confirmed the specificity of the primers. Serum and tissue samples collected from cattle, buffaloes and experimentally infected guinea pigs and calves were subjected to PCR using above primers as well as conventionally used primers G1/G2. All the sera and tissue samples, whether field samples or collected from experimentally infected animals, found positive for G1/G2 specific PCR were also positive for lipL21 and lipL32 specific PCR. The present study indicated that lipL21 and lipL32 based primers could be used for PCR based diagnosis of leptospirosis. Since G1/G2 primers are known not to amplify the DNA of Grippotyphosa, the use of primers employed in the present study could have an additional advantage in detection of cases of the disease.

Cloning and sequence analysis of the gene encoding LipL32 of Leptospira interrogans serovar Sejroe.

Leptospira, a member of the order Spirochaetales, is the causative agent of leptospirosis, an important zoonosis encountered worldwide. The Leptospira interrogans serovar Sejroe was grown in EMJH medium and its DNA was isolated using standard techniques. The LipL32 gene was amplified using the reported primer of Kirschneri of LipL32. The amplified product was found to comprise 756 base pairs. This amplified gene fragment of LipL32 lipoprotein was cloned in E. coli (DH5 alpha) cells using pDrive plasmid as a vector. The recombinant cells were selected on LB agar medium containing ampicillin, X-gal and isopropyl-beta-D-thiogalactopyranoside. Plasmid was extracted from the recombinant white colonies, and restriction endonuclease (RE) analysis was carried out using PstI and SalI. On partial sequence analysis, the product exhibited 756 base pairs, corresponding to 251 amino acids. The cloned gene could be further used for expression of recombinant protein for serodiagnosis of leptospirosis.

Evaluation of a recombinant LipL41 antigen of Leptospira interrogans serovar canicola in ELISA for serodiagnosis of bovine leptospirosis.

The efficacy of a recombinant leptospiral lipoprotein LipL41 as an antigen for conducting enzyme-linked immunosorbent assay (ELISA) for diagnosis of bovine leptospirosis was evaluated. Using known positive and known negative cattle sera the recombinant antigen was found to be highly reactive in the concentration of 100 ng/well. Using a total of 321 field cattle sera the sensitivity of ELISA as compared to microscopic agglutination test (MAT) was calculated to be 100% whereas the specificity was 85.3%. The seropositivity of leptospirosis among bovine population was found to be 21.18% having the predominance of serovars Sejroe and Pomona. It was concluded that rLipL41 protein could be a putative diagnostic candidate for serodiagnosis of bovine leptospirosis.

Immunoreactive outer membrane proteins of Leptospira interrogans serovar Canicola strain Hond Utrecht IV.

BACKGROUND & OBJECTIVES: Leptospirosis is a severe and complex zoonotic disease prevalent in many countries including India. Current leptospiral research is focussed on the identification of the outer membrane proteins (OMPs) of the organism that could be used in developing diagnostic assays for leptospirosis. METHODS: The Leptospira interrogans serovar Canicola was grown in EMJH medium and the cells were subjected to sarcosyl detergent treatment. The sarcosyl soluble (SS) and sarcosyl insoluble (SI) fractions were analyzed by SDS-PAGE and immunoblotting to deduce their protein profile and identifying various immunodominant antigens. RESULTS: The protein profile of SS fractions indicated the presence of three major bands of 41, 32 and 25 kDa and minor bands of 85 and 46 kDa. The SI fraction in serovar Canicola revealed the presence of 112, 93, 77, 43, 36, 29 and 22.5 kDa as major bands and minor bands of 102 and 53 kDa. In immunoblotting, the SS proteins of 41, 32 and 25 kDa and SI proteins of 112, 77, 36 and 22.5 kDa were detected to be major immunogenic proteins. INTERPRETATION & CONCLUSION: In our study immunogenic proteins were extracted from SS and SI fractions and OMPs were similar to those reported in other pathogenic Leptospira strains. These OMPs being unique to all the pathogenic leptospires, can be targeted for diagnostic purpose. Further analysis of the cellular location and expression of leptospiral proteins will be useful in the annotation of genomic sequence data and in providing insight into the biology of Leptospira cells.

Production of ethanol from liquefied cassava starch using co-immobilized cells of Zymomonas mobilis and Saccharomyces diastaticus.

Co-immobilized cells of Saccharomyces diastaticus and Zymomonas mobilis produced a high ethanol concentration compared to immobilized cells of S. diastaticus during batch fermentation of liquefied cassava starch. The co-immobilized cells produced 46.7 g/l ethanol from 150 g/l liquefied cassava starch, while immobilized cells of yeast S. diastaticus produced 37.5 g/l ethanol. The concentration of ethanol produced by immobilized cells was higher than that by free cells of S. diastaticus and Z. mobilis in mixed-culture fermentation. In repeated-batch fermentation using co-immobilized cells, the ethanol concentration increased to 53.5 g/l. The co-immobilized gel beads were stable up to seven successive batches. Continuous fermentation using co-immobilized cells in a packed bed column reactor operated at a flow rate of 15 ml/h (residence time, 4 h) exhibited a maximum ethanol productivity of 8.9 g/l/h.