The high frequency of non-aspartic acid residues at HA222 in influenza A(H1N1) 2009 pandemic viruses is associated with mortality during the upsurge of 2015: a molecular and epidemiological study from central India.

Influenza A(H1N1) viruses of the 2009 pandemic (A(H1N1)pdm09) continue to cause outbreaks in the post-pandemic period. During January to May 2015, an upsurge of influenza was recorded that resulted in high fatality in central India. Genetic lineage, mutations in the hemagglutinin (HA) gene and infection by quasi-species are reported to affect disease severity. The objective of this study is to present the molecular and epidemiological trends during the 2015 influenza outbreak in central India. All the referred samples were subjected to qRT-PCR for diagnosis. HA gene sequencing (23 survivors and 24 non-survivors) and cloning were performed and analyzed using Molecular Evolutionary Genomic Analyzer (MEGA 5·05). Of the 3625 tested samples, 1607 (44·3%) were positive for influenza A(H1N1)pdm09, of which 228 (14·2%) individuals succumbed to death. A significant trend was observed in positivity (P = 0·003) and mortality (P < 0·0001) with increasing age. The circulating A(H1N1)pdm09 virus was characterized as belonging to clade-6B. Clinically significant mutations were detected. Patients infected with the quasi-species of the virus had a greater risk of death (P = 0·009). This study proposes a robust molecular and clinical surveillance program for the detection and characterization of the virus, along with prompt treatment protocols to prevent outbreaks.

Maiden outbreaks of dengue virus 1 genotype III in rural central India.

Dengue is regarded as the most important arboviral disease. Although sporadic cases have been reported, serotypes responsible for outbreaks have not been identified from central India over the last 20 years. We investigated two outbreaks of febrile illness, in August and November 2012, from Korea district (Chhattisgarh) and Narsinghpur district (Madhya Pradesh), respectively. Fever and entomological surveys were conducted in the affected regions. Molecular and serological tests were conducted on collected serum samples. Dengue-specific amplicons were sequenced and phylogenetic analyses were performed. In Korea and Narsinghpur districts 37·3% and 59% of cases were positive, respectively, for dengue infection, with adults being the worst affected. RT-PCR confirmed dengue virus serotype 1 genotype III as the aetiology. Ninety-six percent of infections were primary. This is the first time that dengue virus 1 outbreaks have been documented from central India. Introduction of the virus into the population and a conducive mosquitogenic environment favouring increased vector density caused the outbreak. Timely diagnosis and strengthening vector control measures are essential to avoid future outbreaks.

Emergence of dengue in tribal villages of Mandla district, Madhya Pradesh, India.

BACKGROUND & OBJECTIVES: Dengue (DEN) is a rapidly spreading arboviral disease transmitted by Aedes mosquitoes. Although it is endemic in India, dengue virus (DENV) infection has not been reported from tribal areas of Madhya Pradesh. Investigations were conducted to establish the aetiology of sudden upsurge of cases with febrile illness in June 2013 from tribal villages of Mandla district of Madhya Pradesh, India. METHODS: The rapid response team of the National Institute for Research in Tribal Health, Jabalpur, conducted clinical investigations and field surveys to collect the samples from suspected cases. Samples were tested using molecular and serological tools. Collected mosquitoes were identified and tested for the presence of virus using semi nested reverse transcriptase-polymerase chain reaction (nRT-PCR). The sequences were analysed to identify serotype and genotype of the virus. RESULTS: Of the 648 samples collected from 18 villages of Mandla, 321 (49.53%) were found to be positive for dengue. The nRT-PCR and sequencing confirmed the aetiology as dengue virus type 2. Eighteen per cent of patients needed hospitalization and five deaths were attributed to dengue. The virus was also detected from Aedes aegypti mosquito, which was incriminated as a vector. Phylogenetic analysis revealed that the dengue virus 2 detected belonged to cosmopolitan genotype of the virus. INTERPRETATION & CONCLUSIONS: Dengue virus serotype 2 was detected as the aetiological agent in the outbreak in tribal villages of Mandla district of Madhya Pradesh. Conducive man-made environment favouring mosquitogenic conditions and seeding of virus could be the probable reasons for this outbreak. Urgent attention is needed to control this new threat to tribal population, which is already overburdened with other vector borne diseases.

Circulation of hepatitis A genotype IIIA virus in paediatric patients in central India.

Hepatitis A virus (HAV) infection, a major cause of childhood hepatitis is transmitted by orofaecal route. Children mostly suffer with subclinical infection but may have serious clinical implications leading to hospitalization and mortality. IgM ELISA and nRT PCR were conducted on the blood samples collected from HAV suspected paediatric cases referred to the viral diagnostic laboratory in the Regional Medical Research Centre for Tribals at Jabalpur, Central India. The nRT PCR products were sequenced and phylogenetic analysis was done. Of the 195 samples tested, 41 (21%) were positive for HAV antibodies, among which 38 (92%) belonged to paediatric age group and 32 per cent of these were hospitalized. nRT PCR and sequencing confirmed the presence of HAV. Phylogenic analysis revealed circulation of genotype III A in central India. Regular serological and molecular monitoring would aid in understanding epidemiology of HAV and plan intervention strategies.

Simulated physiological oocyte maturation (SPOM) improves developmental competence of in vitro produced goat embryos.

The effect of simulated physiological oocyte maturation on the developmental competence, reactive oxygen species production and apoptosis rate of in vitro produced goat embryos were studied in the present experiment. Oocytes and spermatozoa were recovered from ovaries and epididymis, respectively, procured from a local small animal abattoir. The oocytes aspirated from the ovaries were allocated into two groups, control (subjected to routine in vitro maturation, fertilization and culture) and simulated physiological oocyte maturation (SPOM) group (subjected to prematuration, followed by routine in vitro maturation, fertilization and culture). The SPOM group showed a significantly (p < 0.05) higher maturation and blastocyst rates (90.60 ± 0.46% and 29.09 ± 2.59%, respectively) as compared to the control group (85.29 ± 0.98% and 24.09 ± 1.08%). The intensity of reactive oxygen species of the embryos in the control group (14.98 ± 0.83 pixels/embryo) was significantly (p < 0.05) higher than the SPOM group (9.60 ± 0.76 pixels/embryo). The apoptosis rate was also significantly (p < 0.05) higher in the embryos of the control group (9.18 ± 1.07%) as compared to the SPOM group (5.71 ± 0.90%). In conclusion, the simulated physiological oocyte maturation system significantly increases the developmental competence of the oocytes and decreases the intensity of reactive oxygen species and embryonic apoptosis in abattoir derived goat embryos.

Responses of cyanobacterium Anabaena doliolum during nickel stress.

Growth and biochemical responses of heterocystous nitrogen fixing cyanobacteria Anabaena doliolum were studied upon exposure to various concentrations of nickel (0.1 to 100 microM) for duration of up to 4 days, in view of its tolerance. The growth of the cyanobacterium was increased in terms of cell density (O.D), heterocyst frequency chlorophyll-a, carotenoid and c-phycocynin up to moderate exposure (10 microM for 96 hr of Ni treatment). Protein and total non protein thiol were measured as stress responsive metabolites and frequency of heterocyst and spores were observed in responses to Ni. The present study demonstrated that the tested cyanobacterium exhibited dose specific responses of metal towards studied different morphological, physiological and biochemical parameters.

Hydration-dependent structural deformation of guanine in the electronic singlet excited state.

Theoretical study was performed to investigate how the degree of hydration affects the structures and properties of the canonical form (keto-N9H) of guanine in the ground and lowest singlet pipi* excited state. This work is the continuation of our earlier work where we have studied the hydration of guanine in the first solvation shell with one, three, five, and six water molecules. In the present investigation, we have considered 7-13 water molecules in hydrating guanine. Ground-state geometries were optimized at the Hartree-Fock level, whereas the configuration interaction-singles (CIS) method was used for the excited-state geometry optimization. The 6-311G(d,p) basis set was used in all calculations. The harmonic vibrational frequency analysis was used to determine the nature of the optimized ground- and excited-state potential energy surfaces; all geometries were found to be minima at the respective potential surfaces. It was found that the degree of hydration has a significant influence on the excited-state structural nonplanarity of guanine. It is expected that excited-state dynamics of guanine will depend on the degree of hydration. Ground- and excited-state geometries of selected hydrated species were also optimized in the bulk water solution using the polarizable continuum model (PCM). It was found that bulk water solution generally does not have significant influence on the structure of the hydrated species. Effects of hydration on different stretching vibrations in the ground and excited states are also discussed.

Role of blue green algae biofertilizer in ameliorating the nitrogen demand and fly-ash stress to the growth and yield of rice (Oryza sativa L.) plants.

Rice is a major food crop throughout the world; however, accumulation of toxic metals and metalloids in grains in contaminated environments is a matter of growing concern. Field experiments were conducted to analyze the growth performance, elemental composition (Fe, Si, Zn, Mn, Cu, Ni, Cd and As) and yield of the rice plants (Oryza sativa L. cv. Saryu-52) grown under different doses of fly-ash (FA; applied @ 10 and 100 tha(-1) denoted as FA(10) and FA(100), respectively) mixed with garden soil (GS) in combination with nitrogen fertilizer (NF; applied @ 90 and 120 kg ha(-1) denoted as NF(90) and NF(120), respectively) and blue green algae biofertilizer (BGA; applied @ 12.5 kg ha(-1) denoted as BGA(12.5)). Significant enhancement of growth was observed in the plants growing on amended soils as compared to GS and best response was obtained in amendment of FA(10)+NF(90)+BGA(12.5). Accumulation of Si, Fe, Zn and Mn was higher than Cu, Cd, Ni and As. Arsenic accumulation was detected only in FA(100) and its amendments. Inoculation of BGA(12.5) caused slight reduction in Cd, Ni and As content of plants as compared to NF(120) amendment. The high levels of stress inducible non-protein thiols (NP-SH) and cysteine in FA(100) were decreased by application of NF and BGA indicating stress amelioration. Study suggests integrated use of FA, BGA and NF for improved growth, yield and mineral composition of the rice plants besides reducing the high demand of nitrogen fertilizers.

Electronic spectra, excited state structures and interactions of nucleic acid bases and base assemblies: a review.

A comprehensive review of recent theoretical and experimental advances in the singlet electronic transitions, excited state structures and dynamics of nucleic acid bases (NABs) and base assemblies are presented. It is well known that NABs absorb ultraviolet radiation, but the absorbed energy is efficiently dissipated in the form of ultrafast internal conversion processes believed to occur in the subpicosecond time scale and, therefore, enabling NABs highly photostable. It is not known how much evolutionary role was played in evolving these molecules and the ultimate selection by nature as genetic materials, but it is well accepted that survival-of-fittest prevails. Recently, significant efforts have been continuously paid to understand the mechanism of electronic excitation deactivation, but universally acceptable mechanism is still elusive. However, recent investigations reveal that electronic excited state geometries of DNA bases are usually nonplanar and this structural nonplanarity may facilitate nonradiative deactivation. Investigation of excited state structures is challenging and, therefore, it is not surprising that despite the impressive theoretical and computational advances, this research area is still hampered by the methodological and computational limitations. Further, stacking has significant influence on the emission properties of molecules. The 2-aminopurine, a fluorescent adenine derivative frequently used in studying DNA dynamics, shows significant attenuations in fluorescence quantum yield when incorporated in the DNA. Theoretical and computational bottlenecks limit a thorough theoretical understanding of effect of stacking interactions on the excited state dynamics of NABs. Despite these limitations the investigations of excited state properties are progressing in the right direction and our better understanding of excited state structure and dynamics of NABs and nucleic acids may help to design preventive strategy for radiation induced illness and photostable materials.

Growth performance and biochemical responses of three rice (Oryza sativa L.) cultivars grown in fly-ash amended soil.

The disposal of fly-ash (FA) from coal-fired power stations causes significant economic and environmental problems. Use of such contaminated sites for crop production and use of contaminated water for irrigation not only decreases crop productivity but also poses health hazards to humans due to accumulation of toxic metals in edible grains. In the present investigation, three rice cultivars viz., Saryu-52, Sabha-5204, and Pant-4 were grown in garden soil (GS, control) and various amendments (10%, 25%, 50%, 75% and 100%) of FA for a period of 90 days and effect on growth and productivity of plant was evaluated vis-a-vis metal accumulation in the plants. The toxicity of FA at higher concentration (50%) was reflected by the reduction in photosynthetic pigments, protein and growth parameters viz., plant height, root biomass, number of tillers, grain and straw weight. However, at lower concentrations (10-25%), FA enhanced growth of the plants as evident by the increase of studied growth parameters. The cysteine and non-protein thiol (NP-SH) content showed increase in their levels up to 100% FA as compared to control, however, maximum content was found at 25% FA in Saryu-52 and Pant-4 and at 50% FA in Sabha-5204. Accumulation of Fe, Si, Cu, Zn, Mn, Ni, Cd and As was investigated in roots, leaves and seeds of the plants. Fe accumulation was maximum in all the parts of plant followed by Si and both showed more translocation to leaves while Mn, Zn, Cu, Ni and Cd showed lower accumulation and most of the metal was confined to roots in all the three cultivars. As was accumulated only in leaves and was not found to be in detectable levels in roots and seeds. The metal accumulation order in three rice cultivars was Fe > Si > Mn > Zn > Ni > Cu > Cd > As in all the plant parts. The results showed that rice varieties Saryu-52 and Sabha-5204 were more tolerant and could show improved growth and yield in lower FA application doses as compared to Pant-4. Thus, Sabha-5204 and Saryu-52 are found suitable for cultivation in FA amended agricultural soils for better crop yields.

Effect of Bradykinin on Murrah buffalo (Bubalus bubalis) semen cryopreservation.

Keeping in view the poor freezability of bubaline semen in conventionally used extenders, this study was conducted on three Murrah bulls to improve semen cryopreservation with the incorporation of Bradykinin (0.5, 1.0 and 2.0 ng ml(-1)) in routinely used egg yolk tris-glycerol (EYTG) extender. Bradykinin (2.0 ng ml(-1)) had significant (P<0.05) beneficial effect on live sperm % (81.6+/-1.8) and hypo osmotic swelling (HOS) % (63.0+/-1.3) as compared to their respective control values of 73.4+/-2.1 and 56.3+/-2.0 at 0 h post freezing. The post-thaw progressive sperm motility in semen samples diluted with EYTG containing 2.0 ng ml(-1) Bradykinin (65.5+/-1.4) was also significantly (P<0.01) higher than control (60.3+/-1.9) at 0 h post freezing. Thus incorporation of 2 ng ml(-1) Bradykinin in buffalo semen diluted in EYTG extender may be useful in improving the quality of cryopreserved bubaline semen.

Vibrational analysis of complexes of urate with IA group metal cations (Li+, Na+ and K+).

Vibrational frequency analysis was performed for the complexes of alkali metal cations (Li+, Na+ and K+) with urate in the gas phase. The geometries of all possible metal cation-urate complexes were optimized at the B3LYP/6-311++G(d,p) level. The most stable complex corresponding to the each cation was used for the vibrational frequency analysis including the computation of % potential energy distribution (%PED). For comparison, the vibrational frequency analysis was also performed for the uric acid. The computed results are discussed in terms of the available experimental data. It was revealed that the characteristic stretching vibrational modes corresponding to the metal cation and the interacting nucleophilic sites of urate can be used to identify metals involved in the stone formation in the living system. Changes in different vibrational frequencies of uric acid consequent to the metal cation interactions are discussed.

Morphology and cultural behavior of Botryococcus protuberans with notes on the genus.

The green alga Botryococcus protuberans was isolated from its natural environment and its morphology under different cultural conditions was examined. The alga was characterized by a high starch content and reddish oil drops as the assimilatory products. Photosynthetic pigments, Chl a, Chl b, carotenoids and xanthophylls are present. Modification of environmental conditions in modified Chu-10 medium resulted in optimum growth of the alga. Fatty acid composition revealed palmitic acid being the major component, while lauric acid, myristic acid and stearic acid were found in less quantity.

In silico kinetics of alkaline hydrolysis of 1,3,5-trinitro-1,3,5-triazinane (RDX): M06-2X investigation.

Alkaline hydrolysis of RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine), as one of the most promising methods for nitrocompound remediation, was investigated computationally at the PCM(Pauling)/M06-2X/6-311++G(d,p) level of theory. Computational simulation shows that RDX hydrolysis is a highly exothermic multistep process involving initial deprotonation and nitrite elimination, cycle cleavage, further transformation of cycle-opened intermediates to end products caused by a series of C-N bond ruptures, hydroxide attachments, and proton transfers. Computationally predicted products of RDX hydrolysis such as nitrite, nitrous oxide, formaldehyde, formate, and ammonia correspond to experimentally observed ones. Accounting of specific hydration of hydroxide is critical to create an accurate kinetic model for alkaline hydrolysis. Simulated kinetics of the hydrolysis are in good agreement with available experimental data. A period of one month is necessary for 99% RDX decomposition at pH 10. Computations predict significant increases of the reaction rate of hydrolysis at pH 11, pH 12, and pH 13.

Effect of hydration on the lowest singlet PiPi* excited-state geometry of guanine: a theoretical study.

An ab-initio computational study was performed to investigate the effect of explicit hydration on the ground and lowest singlet PiPi* excited-state geometry and on the selected stretching vibrational frequencies corresponding to the different NH sites of the guanine acting as hydrogen-bond donors. The studied systems consisted of guanine interacting with one, three, five, six, and seven water molecules. Ground-state geometries were optimized at the HF level, while excited-state geometries were optimized at the CIS level. The 6-311G(d,p) basis set was used in all calculations. The nature of potential energy surfaces was ascertained via the harmonic vibrational frequency analysis; all structures were found minima at the respective potential energy surfaces. The changes in the geometry and the stretching vibrational frequencies of hydrogen-bond-donating sites of the guanine in the ground and excited state consequent to the hydration are discussed. It was found that the first solvation shell of the guanine can accommodate up to six water molecules. The addition of the another water molecule distorts the hydrogen-bonding network by displacing other neighboring water molecules away from the guanine plane.

Comparison of prolactin, CA 15-3 and TPA in breast carcinomas.

Circulating prolactin, CA 15-3 and TPA were assayed pre-therapeutically and sequentially thereafter from 68 breast cancer patients attending the Gujarat Cancer and Research Institute, Ahmedabad--a regional cancer institute in Western India. The three marker values were correlated with the stage, histologic grade and disease status. At least one of the markers was elevated in 82% of patients. CA 15-3 and TPA levels were elevated with the advancement of stage. Prolactin levels were high in poorly differentiated tumors of pre-menopausal patients. The disease status was effectively reflected by the levels of prolactin and CA 15-3. TPA showed high false positivity so was of no use as an indicator of disease status. Recurrence could be predicted early, with a lead time of 3-6 months using prolactin and CA 15-3.

Prognostic value of squamous cell carcinoma antigen in patients with uterine cervical carcinoma.

Squamous cell carcinoma antigen (SCC Ag) was estimated in 30 controls, in 16 patients with benign lesions of the uterine cervix, and in 51 patients with uterine cervical carcinoma. The rate of positivity of the antigen among the cancer patients was 87% (N = 49). SCC Ag estimations were of no diagnostic value, since 37% of patients with benign lesions had elevated levels compared to controls. SCC Ag was highly correlated to histologic subtype. The highest values were obtained in keratinizing tumors, followed by large cell nonkeratinizing and small cell nonkeratinizing types. Moreover, elevation of SCC Ag was statistically significant (p less than 0.001) among all the three histologic subtypes compared to controls. However, SCC Ag levels were not consistently correlated to the stage of the disease. Patients with pretherapeutic SCC Ag levels above 30 ng/ml had a faster recurrence rate and shorter survival than those who exhibited antigen values below 2.0 ng/ml. From our results, it is suggested that SCC Ag has limited use as a parameter for early diagnosis of cervical carcinoma, but it seems to reflect advancement of the disease. These findings indicate that SCC Ag elevation may prove to be a valuable marker in predicting subclinical disease.

Electronic spectra of adenine and 2-aminopurine: an ab initio study of energy level diagrams of different tautomers in gas phase and aqueous solution.

Ground and lowest two singlet excited state geometries of four tautomeric forms (N9H, N7H, N3H and N1H) of each of adenine and 2-aminopurine (2AP) were optimized using an ab initio approach employing a mixed basis set (6-311 + G* on the nitrogen atom of the amino group and 4-31G basis set on the other atoms). Excited states were generated employing configuration interaction involving single electron excitations (CIS). Subsequently, the different species were solvated in water employing the self-consistent reaction field (SCRF) approach along with the corresponding gas phase optimized geometries. Thus the observed absorption and fluorescence spectra of adenine and 2AP have been explained successfully. It is concluded that both the N9H and N7H forms of 2AP would contribute to absorption and fluorescence spectra. Further, the fluorescence of 2AP would be absorbed by its cation in which both the N9 and N7 atoms are protonated, the fluorescence of which can have an anti-Stokes component. Among the different tautomers of adenine, the N9H form would be present dominantly in the ground state in aqueous solutions but the N7H form would be produced by energy transfer and subsequent fluorescence. The N3H form of adenine appears to be responsible for the observed absorption near 300 nm by its solutions intermittently exposed to ultraviolet radiation. The rings of the different species related to 2AP and adenine remain almost planar in the pi-pi* and n-pi* singlet excited states as in the ground state. The pyramidal character of the amino group is usually less in the pi-pi* excited states than that in the corresponding ground or n-pi* excited states. Molecular electrostatic potential (MEP) maps of the molecules provide useful clues regarding phototautomerism.

A geometry optimization and molecular electrostatic potential mapping study of structure-activity relationship for some anti-Alzheimer agents.

Molecular geometries of some substituted (pyrroloamino)pyridines which possess anti-Alzheimer activity were optimized and potential-derived CHelpG point charges were computed using ab initio SCF molecular orbital approach employing the 3-21G basis set. AM1 molecular orbital calculations were performed using these optimized geometries and thus optimized Hybridization. Displacement Charges (HDC) combined with Löwdin charges continuously distributed in three dimension were obtained. Molecular electrostatic potential (MEP) maps of the molecules were obtained in two ways: (i) using the HDC-based model with the help of which MEP minima near the molecules were located, and (ii) using the CHelpG point charges, MEP values on the van der Waals surfaces of the molecules were computed. The MEP maps computed using both the methods have negative MEP regions near the pyridine nitrogen atom which appears to be the main binding site of the molecules with the appropriate receptor. Both electrostatic interaction and lipophilic association between these molecules and the receptor appear to contribute to biological activity.

Constructed wetland as an ecotechnological tool for pollution treatment for conservation of Ganga river.

With aim to develop an efficient and ecofriendly approach for on-site treatment of sewage, a sub-surface flow constructed wetland (CW) has been developed by raising potential aquatic macrophytes; Typha latifolia, Phragmites australis, Colocasia esculenta, Polygonum hydropiper, Alternanthera sessilis and Pistia stratoites in gravel as medium. Sewage treatment potential of CW was evaluated by varying retention time at three different stages of plant growth and stabilization. After 6 months, monitoring of fully established CW indicated reduction of 90%, 65%, 78%, 84%, 76% and 86% of BOD, TSS, TDS, NO3-N, PO4-P and NH4-N, respectively in comparison to inlet after 36 h of retention time. Sewage treatment through CW also resulted in reduction of heavy metal contents. Thus, CW proved an effective method for treatment of wastewater and may be developed along river Ganga stretch as an alternative technology. Treated water may be drained into river to check further deterioration of Ganga water quality.