Impact of effluent parameters and vancomycin concentration on vancomycin resistant Escherichia coli and its host specific bacteriophage lytic activity in hospital effluent.

Vancomycin resistance in bacteria has been classified under high priority category by World Health Organization (WHO) and its presence in hospital effluent is reported to be increasing owing to excess antibiotics use. Among various strategies, bacteriophage has been recently considered as a promising biological agent for combating such antimicrobial resistant bacteria (ARB). However, the influence of effluent's properties on phage-ARB interaction in actual hospital effluent is not completely understood. The present works intends to study this influence of hospital effluent and its parameters on the interaction between vancomycin resistant E. coli (VRE) and its host specific bacteriophage. The isolated VRE was identified by 16S rRNA sequencing, matrix-assisted laser desorption/ionization-time of flight (MALDI - TOF) and whole genome sequencing. The infectivity of phage onto host bacteria was investigated using electron microscopic techniques, dynamic light scattering (DLS), spectrofluorophotometer and confirmed using double agar overlay method. The monovalency and polyvalency of isolated phage against various bacterial species were determined. The phage morphology was identical to T7 phage belonging to Podoviridae. The phage lysis was maximum at pH 7 (90.2%), 37 °C (91.6%) and vancomycin concentration of 50 µg/mL in both synthetic media (89.13%) and effluent (100%). At a maximum vancomycin concentration of 100 µg/mL, decrease in Ca, K, Mg and P (up to 19.70, 14.18, 28, and 15.82% respectively) concentration in effluent was observed due to phage infectivity when compared to control. The whole genome sequencing was performed and the bioinformatics analysis presented the role of mdfA gene encoding the efflux pump in causing vancomycin resistance in E. coli. It also depicted the presence of multiple genes responsible for mercury, cobalt, zinc and cadmium resistance in VRE. These results clearly indicate that bacteriophage mediated combating of VRE is possible in actual hospital effluent and can be used as one of the treatment methods.

Elevated CO2 ameliorates the high temperature stress effects on physio-biochemical, growth, yield traits of maize hybrids.

The rising temperatures and levels of carbon dioxide in the atmosphere are anticipated to have a significant impact on the productivity of agricultural crops. Although, the individual effects of elevated CO2 and temperature have been extensively studied in C3 and C4 crops, there remains a scarcity of research investigating their interactive effects specifically on maize hybrids. The impact of elevated temperature and its interaction with elevated CO2 on phenology, physiology, biomass, and grain yield of maize hybrids was assessed in a field experiment using Free Air Temperature Elevation (FATE) facility. The results showed that elevated temperature (eT) increased the anthesis silking interval (ASI), while the presence of elevated CO2 along with elevated temperature (eT + eCO2) mitigated this effect. The differential expression were observed between hybrids depending on their genetic potential. Furthermore, the net photosynthetic rate (Anet), stomatal conductance (gs), and transpiration rate (Tr) of hybrids decreased under elevated temperature but eT + eCO2 condition helped in reverting its impact to some extent. In term of leaf composition, the highest level of total soluble sugars (TSS) and starch was observed under eT + eCO2 conditions, possibly due to improved Anet in the presence of elevated eCO2. The negative impact of eT was also evident through increased proline and MDA content, but eT + eCO2 ameliorated the adverse effect of eT. The biomass and grain yield also responded similarly, among the hybrids 900M GOLD recorded superior performance for grain yield at eT condition exceeding 35 °C. On the other hand, DHM117 experienced a significant reduction in grain yield under eT, but performed better under eT + eCO2 due to its improved physiological response to eCO2. The study indicated that elevated levels of carbon dioxide can actually mitigate the detrimental effects of elevated temperature on maize crop. This positive impact on maize crop can be attributed to an enhanced physiological performance in the presence of eCO2 which enables the plants to maintain satisfactory yield levels despite the challenging environmental conditions.

A Study on Knowledge, Attitude, and Practice Related to Sexually Transmitted Infections Among Students of Nonmedical Background in Manipal, India.

Background: Sexually transmitted infections (STIs) are one of the major public health problems in India with adolescents being a vulnerable section of the affected population. According to the World Health Organization (WHO), one in twenty adolescents contracts an STI every year, excluding the human immunodeficiency virus (HIV) and other viral infections. It is important to assess the knowledge, attitude, and practices related to STIs among adolescents and young adults of non-medical backgrounds to educate them properly about protective and preventive measures and to allay misconceptions, if any. Materials and Methods: This is a cross-sectional study involving 507 engineering students who were asked to fill out the predesigned anonymous questionnaire form. Results were statistically analyzed and tabulated. Results: Of the 507 students, 375 were males and 132 were females. The mean age of the patients was 20.5 years. HIV was the most commonly identified STI (94.2%), followed by herpes genitalis (76.5%). Around 97.4% and 85.7% identified the sexual route and blood transfusion as the modes of transmission of STIs, respectively. A total of 90.9% said that the Internet was their primary source of information regarding STIs. Infertility and cancer were identified as complications by 59.5% and 25% of them, respectively. Only 23.4% were aware of the National STI Control Program. The results with respect to attitudes and practices related to STIs were variable. Conclusion: Our study reflects that with change in time, the attitude of young adults toward sexuality is becoming very casual, but knowledge remains low. A lot of educational activities should be undertaken to educate our youth about various aspects of STIs and thus reduce the burden of the disease in the community.

Recycled mesoporous magnetic composites with high surface area derived from plastic and de-oiled sludge wastes: An empirical comparison on their competitive performance for toxic Cr (VI) removal.

The current study focused on the comparative removal of hexavalent chromium using two magnetically modified hybrid adsorbent composites. Their precursor chars were prepared from bio sludge and plastic waste derivatives. The coating of magnetite on the chars' surface was evident from the SEM micrographs. Infusion of magnetite nanoparticles in the native chars aided in the reduction of the composite particles' sizes, thus, forming high surface area composites. Screening of uptake capacities among various blends of char and magnetite were surveyed. Composites of both kinds with a 1:5 ratio of char: iron salts composition answered well. The pHZPC and zeta potential values of the composites indicated the neutral charge on the composites' surface. This suggested the need for a highly acidic environment for efficient Cr(VI) removal. Optimum economic conditions for Cr(VI) removal were obtained from the batch studies (solution pH - 1.5; contact time - (a) MPC [magnetic plastic char] - 60 min (b) MBC [magnetic biochar] - 40 min; temperature - 25 °C). The maximum monolayer adsorption capacity of MPC and MBC were found to be 84.67 mg/g and 53.83 mg/g respectively. Isotherm, kinetic and thermodynamic studies revealed the adsorption systems' inclination towards physisorption. From the characterization and modeling results, electrostatic force of attraction and pore filling was anticipated to be the mechanism of adsorption for both MPC and MBC. Thus, in the relative removal studies, MBC was found to compete better than MPC due to its enhanced porosity and surface area.

Intraspecific differences in long-term drought tolerance in perennial ryegrass.

Lolium perenne L. (perennial ryegrass) is the most important pasture grass species in temperate regions of the world. However, its growth is restricted in summer dry environments. Germplasm screening can be used to identify accessions or individual plants for incorporation into breeding programs for drought tolerance. We selected nine perennial ryegrass accessions from different global origins and from a range of climatic and environmental conditions. In addition, the perennial ryegrass cultivar 'Grasslands Impact' was chosen as a reference. The accessions were grown for 360 days in a controlled environment through six consecutive drought stress and recovery cycles. We observed intraspecific differences in drought stress responsiveness for shoot biomass and survival from the third stress cycle. An accession from Norway had 50% more shoot dry matter than the next best-performing accession after six drought cycles. Compared with the reference cultivar 'Grasslands Impact', shoot dry matter of the accession from Norway was more than seven times higher after six drought cycles, indicating superior performance of this ecotype under drought stress. Drought tolerance was characterized by osmotic adjustment and higher relative leaf water content at low soil moisture levels. Furthermore, the findings of this study identify solute potential as an early predictor of drought stress tolerance. These intraspecific differences can be used in breeding programs for the development of drought-tolerant perennial ryegrass cultivars.

Genotypic variability in physiological, biomass and yield response to drought stress in pigeonpea.

Three pigeonpea (Cajanus cajan L. Millsp.) genotypes- GT-1, AKP-1 and PRG-158 with varying crop duration, growth habit and flowering pattern were evaluated for variability in their response for drought stress. Drought stress was imposed at initiation of flowering and the observations on biomass and seed yield parameters were recorded at harvest. The magnitude of response of individual component to drought stress was found to be genotype specific. Drought stress significantly decreased photosynthetic rate (PN), transpiration rate (Tr) and relative water content (RWC) in all the genotypes, however the magnitude of reduction differed with genotype. With drought stress, the reduction of PN was highest in GT-1 while reduction in Tr was highest in PRG-158. The genotype AKP-1, accumulated significantly higher concentrations of osmotic solutes especially proline under water deficit stress, this facilitated it to maintain higher relative water content (RWC) and lower malondialdehyde (MDA) content as compared to other genotypes. Drought stress also impacted biomass production and their partitioning to vegetative and reproductive components at harvest. There was significant variability between the genotypes for seed yield under drought stress while it was non-significant under well-watered condition. Drought stress enhanced flower drop and decreased flower to pod conversion resulting in reduced pod number and seed number in PRG-158 and GT-1. The genotype AKP-1 recorded superior performance for seed yield under stress environment due to its ability in maintaining pod and seed number as well as improved test weight (100 seed weight). Under drought stress, significant positive association of seed yield with proline, seed number, pod number and test weight clearly indicating their role in drought tolerance.

A molecular dynamics study of the interaction of oleate and dodecylammonium chloride surfactants with complex aluminosilicate minerals.

Surface characteristics of complex aluminosilicate minerals like spodumene [LiAl(SiO(3))(2)], jadeite [NaAl(SiO(3))(2)], feldspar [KAlSi(3)O(8)], and muscovite [K(2)Al(4)(Al(2)Si(6)O(20))(OH)(4)]) are modeled. Surface energies are computed for the cleavage planes of these minerals. Adsorption mechanisms of anionic chemisorbing type oleate and cationic physisorbing type dodecylammonium chloride molecules on two different crystal planes, that is (110) and (001), of spodumene and jadeite are studied in terms of the surface-surfactant interaction energies computed using molecular dynamics (MD) simulations. The conclusions drawn from purely theoretical computations match remarkably well with our experimental results.

Molecular dynamic simulations of self-assembled alkylthiolate monolayers on an Au(III) surface.

Molecular dynamics simulations incorporating explicit gold atoms in the simulations have been carried out for alkanethiol self-assembled monolayers chemisorbed on the Au(III) surface. The structural properties of the monolayer are evaluated for two force fields: one in which the Au--S--C bond is fixed (FF I), and the other in which it is flexible (FF II). The influence of these force fields on the structural properties of HS(CH2)14CH3 on the structured Au surface is compared at different temperatures. FF I yields greater tilt angles and a smaller film thickness when compared with FF II. Both of the force fields predict that the tilt angles do not follow a monotonic decrease with temperature but show minima around 200 K. Simulations carried out at different chain lengths at 300 K reveal that FF II predicts a greater film thickness than FF I; however, the difference is within 1 A.

Starch branching enzymes in sorghum (Sorghum bicolor) and barley (Hordeum vulgare): comparative analyses of enzyme structure and gene expression.

A genomic clone for starch branching enzyme (SBE) IIb was isolated from a sorghum bacterial artificial chromosome (BAC) library. The promoter and 5' flanking sequence, the first four exons and introns as well as the last exon and the 3' untranslated region were sequenced. The tentative transcription start site of sorghum sbeIIb was mapped based on alignment with the maize sbeIIb gene. The exon-intron structure of the 5' portion of sorghum sbeIIb was similar to that of maize sbeIIb but differed from that of barley sbeIIb. Specifically, the intronic BbI element involved in the endosperm specific expression of barley sbeIIb was lacking in the sorghum gene. A cDNA clone for sorghum sbeIIb was reverse PCR amplified and found to encode an 803 amino acids long protein. The amino acid sequence of sorghum SBEIIb was compared to that of sorghum SBEI and corresponding enzymes in barley. The overall identity in amino acid sequence was 54% in the central portion of the enzymes. A major difference between the SBEII and SBEI isoforms was a 67 amino acids-long C-terminal extension in the SBEIs. The spatial and temporal expression patterns of sorghum sbeIIb was determined and compared with those of the sorghum gene for SBEI and the barley genes for SBEIIB and SBEI. All four genes exhibited a seed specific expression. However, while barley sbeIIb and sbeI transcripts were detected exclusively in the endosperm, the sorghum genes were expressed also in the embryo. The activity of sorghum sbeIIb and sbeI exhibited a late onset, with a peak of transcription at around 22 days after pollination. This is similar to the pattern of barley sbeI but different from that of barley sbeIIb, which showed a peak of transcription at 12 days after pollination.

Expression of thaumatin-like permatin PR-5 genes switches from the ovary wall to the aleurone in developing barley and oat seeds.

Permatins are antifungal thaumatin-like proteins (TLPs) of the PR-5 family of pathogenesis-related proteins. They occur in many cereals, but little is known of their expression and roles. Permatin cDNA clones were produced and used to study expression in developing barley and oat seeds. Actin and CDC48 mRNAs declined rapidly following inoculation of barley spikes with Fusarium graminearum. Despite this, permatin mRNA levels remained constant or increased slightly. Studies of permatin gene expression in healthy plants revealed that developing barley and oat seeds accumulate permatin mRNA in an unusual bimodal pattern. Permatin mRNA and protein are highly abundant around the time of pollination and then decrease rapidly to near-zero. A second peak occurs in the doughy stage of development. Antibody and DNA probe hybridization studies showed that expression initially occurs in the ovary wall and then switches to the aleurone and ventral furrow of developing seeds, reaching a peak in the doughy stage. Small amounts of permatin mRNAs also occur in certain vegetative tissues. The barley and oat permatin sequences provided sufficient comparisons between cereal TLPs to suggest that deletions or additions in specific elements could have led to the divergence of leaf- and seed-specific TLPs.

Analyses of isoamylase gene activity in wild-type barley indicate its involvement in starch synthesis.

The notion of debranching enzyme activity as a participant in starch synthesis is gaining acceptance. Inconsistent reports from mutant analyses implicate either isoamylase or pullulanase as a determinant in amylopectin formation and whether wild-type plants utilize one or the other, or both, of these debranching enzymes in starch synthesis is unclear. Recent results on the sul mutant in maize suggest that both forms of debranching enzymes might be involved in amylopectin formation. We wished to find out if isoamylase takes part in starch synthesis by comparing isoamylase gene activity under three conditions: (1) during starch accumulation in developing sink tissues; (2) during starch degradation in germinating seeds; (3) in ectopic expression after applying sucrose, a starch precursor. We isolated the gene for barley isoamylase, isol, and analysed its expression and regulation in germinating seeds, developing endosperm and vegetative tissues, and compared the isoamylase gene expression in sink tissues from three different species. Our results indicate that isoamylase gene activity is involved in starch synthesis in wild-type plants and is modulated by sucrose.

The two genes encoding starch-branching enzymes IIa and IIb are differentially expressed in barley.

The sbeIIa and sbeIIb genes, encoding starch-branching enzyme (SBE) IIa and SBEIIb in barley (Hordeum vulgare L.), have been isolated. The 5' portions of the two genes are strongly divergent, primarily due to the 2064-nucleotide-long intron 2 in sbeIIb. The sequence of this intron shows that it contains a retro-transposon-like element. Expression of sbeIIb but not sbeIIa was found to be endosperm specific. The temporal expression patterns for sbeIIa and sbeIIb were similar and peaked around 12 d after pollination. DNA gel-blot analysis demonstrated that sbeIIa and sbeIIb are both single-copy genes in the barley genome. By fluorescence in situ hybridization, the sbeIIa and sbeIIb genes were mapped to chromosomes 2 and 5, respectively. The cDNA clones for SBEIIa and SBEIIb were isolated and sequenced. The amino acid sequences of SBEIIa and SBEIIb were almost 80% identical. The major structural difference between the two enzymes was the presence of a 94-amino acid N-terminal extension in the SBEIIb precursor. The (beta/alpha)8-barrel topology of the alpha-amylase superfamily and the catalytic residues implicated in branching enzymes are conserved in both barley enzymes.

Identification of four starch-branching enzymes in barley endosperm: partial purification of forms I, IIa and IIb.

Developing caryopses of barley (Hardeum vulgare L. ev. Golf) were monitored for starch-branching enzyme activity for 1 month after anthesis. Homogenized caryopses from grains with the highest specific activity were used as starting material for purification of branching enzyme by FPLC chromatography. Several branching enzyme activity fractions were resolved. We have previously described the isolation of a 51/50 kDa protein with starch branching activity from one of these fractions. From three other fractions, we have now partially purified starch branching enzymes corresponding to forms I, IIa and IIb.

Rice anther culture: callus initiation and androclonal variation in progenies of regenerated plants.

Anthers from rice (Oryza sativa L.) subspecies japonica initiated more callus than their indica or indica x japonica counterparts. A mild stress, either by slow desiccation or heat shock, prior to the plating of anthers enhanced the ability to initiate callus. Slow dessication of anthers enhanced the ability of the japonica anthers to initiate callus even in medium that was supplemented with NaCl. The ability to initiate callus by the anthers plated on NaCl-supplemented medium decreased as the NaCl level in the medium increased. Among the regenerated plants 2.5% were albino and another 2% were haploid. Androclonal variation for tiller numbers, shoot height, plant dry matter and flowering were noticed in the progenies of the regenerated plants.

Plant regeneration from in vitro cultures of anthers and mature seeds of rice (Oryza sativa L.) cv. Basmati-370.

Pollen plants were obtained from anther-derived calli of the indica rice variety Basmati-370. Anther-response (anthers producing pollen derived calli) and plant regeneration frequency from the pollen derived calli. was very low. Donor plants which flowered at the average max/min. temperature of 34.2°/23.3°C gave a significantly higher anther-response to in vitro techniques, than did those which flowered at 29.1°/16.4°C. Somatic callus induction and subsequent plant regeneration was readily obtained from mature seed embryos. While 2,4-D or 2,4,5-T (1 or 2 mg/l) proved highly efficient for callus induction, tryptophan (50 or 100 mg/l) induced a high frequency of green plants from the calli.