Full aperture backscatter diagnostics for characterization of laser plasma instabilities at the extreme light infrastructure (ELI) beamlines.

We report on the commissioning of a full aperture backscatter diagnostics station for the kilojoule, nanosecond high repetition rate L4n laser operating at a wavelength of 527 nm at the Extreme Light Infrastructure (ELI) - Beamlines, Dolni Brezany, Czech Republic. Light scattered back from laser-plasma interaction into the cone of the final focusing lens is captured and split into different channels to measure the signatures of laser plasma instabilities from stimulated Brillouin scattering, stimulated Raman scattering, and two plasmon decay with respect to back scattered energy, its spectrum, and its temporal profile. The performance was confirmed in a commissioning experiment with more than 800 shots at laser intensities ranging from 0.5 × 1013 to 1.1 × 1015 W cm-2. These diagnostics are permanently installed at ELI Beamlines, and can be used to understand the details of laser-plasma interactions in experiments with kJ and 527 nm light. The large number of shots that can be collected in an experimental campaign will allow us to study the details of the laser-plasma interaction with a high level of confidence.

Counterpropagating Radiative Shock Experiments on the Orion Laser.

We present new experiments to study the formation of radiative shocks and the interaction between two counterpropagating radiative shocks. The experiments are performed at the Orion laser facility, which is used to drive shocks in xenon inside large aspect ratio gas cells. The collision between the two shocks and their respective radiative precursors, combined with the formation of inherently three-dimensional shocks, provides a novel platform particularly suited for the benchmarking of numerical codes. The dynamics of the shocks before and after the collision are investigated using point-projection x-ray backlighting while, simultaneously, the electron density in the radiative precursor was measured via optical laser interferometry. Modeling of the experiments using the 2D radiation hydrodynamic codes nym and petra shows very good agreement with the experimental results.

Amyotrophic Lateral Sclerosis and Metabolomics: Clinical Implication and Therapeutic Approach.

Amyotrophic lateral sclerosis (ALS) is one of the most common motor neurodegenerative disorders, primarily affecting upper and lower motor neurons in the brain, brainstem, and spinal cord, resulting in paralysis due to muscle weakness and atrophy. The majority of patients die within 3-5 years of symptom onset as a consequence of respiratory failure. Due to relatively fast progression of the disease, early diagnosis is essential. Metabolomics offer a unique opportunity to understand the spatiotemporal metabolic crosstalks through the assessment of body fluids and tissue. So far, one of the most challenging issues related to ALS is to understand the variation of metabolites in body fluids and CNS with the progression of disease. In this paper we will review the changes in metabolic profile in response to disease progression condition and also see the therapeutic implication of various drugs in ALS patients.

Modulation of Bax/Bcl-2 and caspases by probiotics during acetaminophen induced apoptosis in primary hepatocytes.

Oxidative stress is an important factor in drug induced hepatotoxicity and antioxidants from natural sources have potential to ameliorate it. The present study was aimed to investigate cyto-protective potential of probiotic Enterococcus lactis IITRHR1 (El(SN)) and Lactobacillus acidophilus MTCC447 (La(SN)) lysate against acetaminophen (APAP) induced hepatotoxicity. Cultured rat hepatocytes pretreated with El(SN)/La(SN) showed higher cell viability under APAP stress. Pre-treatment with El(SN,) restored glutathione level and reduced ROS generation significantly which are major biomarkers of oxidative stress. It also reduced NO level, MDA formation and enhanced SOD activity. Pre-treatment with probiotic lysates significantly inhibited the translocation of pro-apoptotic protein (Bax), enhanced anti-apoptotic (Bcl-2) protein levels and prevented release of cyt c to cytosol; suggesting involvement of mitochondrial proteins in protection against APAP induced oxidative cellular damage. Loss in mitochondrial membrane potential due to APAP treatment was prevented in the presence of probiotic lysates. Protective action of El(SN)/La(SN) pretreatment was further supported by prevention of procaspase-3 activation, DNA fragmentation and chromatin condensation, in turn inhibiting APAP induced apoptotic cell death. The results indicate that probiotic preparations modulate crucial end points of oxidative stress induced apoptosis and may be used for management of drug induced liver injury.

Metabolomic analysis of serum by (1) H NMR spectroscopy in amyotrophic lateral sclerosis.

BACKGROUND: Amyotrophic lateral sclerosis (ALS), an invariably fatal neurological disorder shows complicated pathogenesis that poses challenges with respect to diagnosis as well as monitoring of disease progression. METHODS: We investigated metabolite profiles in the serum of 30 patients with ALS, 10 patients of Hirayama disease, which served as a neurological disease control and 25 healthy controls by using (1) H NMR spectroscopy. RESULTS: Compared to healthy controls, the ALS patients had higher quantities of glutamate (P<0.001), beta-hydroxybutyrate (P<0.001), acetate (P<0.01), acetone (P<0.05), and formate (P<0.001), and lower concentrations of glutamine (P<0.02), histidine (P<0.001) and N-acetyl derivatives. On the other hand, Hirayama disease patients had significantly higher median concentrations of pyruvate (P<0.05), glutamate (P<0.001), formate (P<0.05) and lower median concentrations of N-acetyl derivatives. Furthermore, we also found that serum glutamate showed a positive correlation (P<0.001, r=0.6487) whereas, histidine showed a negative correlation (P<0.001, r=-0.5641) with the duration of the disease in ALS. CONCLUSIONS: Such (1) H NMR study of serum may reveal abnormal metabolite patterns, which could have the potential to serve as surrogate markers for monitoring ALS disease progression.

Oxidative DNA damage protective activity, antioxidant and anti-quorum sensing potentials of Moringa oleifera.

The aqueous extract of leaf (LE), fruit (FE) and seed (SE) of Moringa oleifera was assessed to examine the ability to inhibit the oxidative DNA damage, antioxidant and anti-quorum sensing (QS) potentials. It was found that these extracts could significantly inhibit the OH-dependent damage of pUC18 plasmid DNA and also inhibit synergistically with trolox, with an activity sequence of LE > FE > SE. HPLC and MS/MS analysis was carried out, which showed the presence of gallic acid, chlorogenic acid, ellagic acid, ferulic acid, kaempferol, quercetin and vanillin. The LE was with comparatively higher total phenolics content (105.04 mg gallic acid equivalents (GAE)/g), total flavonoids content (31.28 mg quercetin equivalents (QE)/g), and ascorbic acid content (106.95 mg/100 g) and showed better antioxidant activity (85.77%), anti-radical power (74.3), reducing power (1.1 ascorbic acid equivalents (ASE)/ml), inhibition of lipid peroxidation, protein oxidation, OH-induced deoxyribose degradation, and scavenging power of superoxide anion and nitric oxide radicals than did the FE, SE and standard alpha-tocopherol. Eventually, LE and FE were found to inhibit violacein production, a QS-regulated behavior in Chromobacterium violaceum 12472.

Polyphenolics from various extracts/fractions of red onion (Allium cepa) peel with potent antioxidant and antimutagenic activities.

In order to determine antioxidant activity, the five extracts/fractions of red onion peel were studied for their total content of phenolics (TPC), flavonoids (TFC), antioxidant activity (AOA), free radical scavenging activity (FRSA), assayed by DPPH radical in the terms of anti-radical power (ARP) and reducing power (RP), expressed as ascorbic acid equivalents (ASE)/ml. High TPC (384.7 +/- 5.0 mg GAE/g), TFC (165.2+/- 3.2 mg QE/g), AOA (97.4 +/- 7.6%), ARP (75.3 +/-4.5) and RP (1.6 +/-0.3 ASE/ml) were found for the ethyl acetate (EA) fraction. EA fraction had markedly higher antioxidant capacity than butylated hydroxytoluene (BHT) in preventive or scavenging capacities against FeCl3-induced lipid peroxidation, protein fragmentation, hydroxyl (site-specific and non-site-specific), superoxide anion and nitric oxide radicals. EA fraction also showed dose dependent antimutagenic activity by following the inhibition of tobacco-induced mutagenicity in Salmonella typhimurium strains (TA102) and hydroxyl radical-induced nicking in plasmid pUC18 DNA. HPLC and MS/MS analysis showed the presence of ferulic, gallic, protocatechuic acids, quercetin and kaempferol. The large amount of polyphenols contained in EA fraction may cause its strong antioxidant and antimutagenic properties. This information shows that EA fraction of red onion peel can be used as natural antioxidant in nutraceutical preparations.

Antioxidant and anti-quorum sensing activities of green pod of Acacia nilotica L.

The antioxidant and anti-quorum sensing activities of eight extracts were studied in green pods of Acacia nilotica. The specific phenolic compositions and their quantifications were performed by HPLC and MS/MS, which showed that the HEF (pH 4) was higher in gallic acid, ellagic acid, epicatechin, rutin, and GTs. In order to find antioxidant potential of various extracts, their activities were studied for TPC, AOA, FRSA, RP, inhibition of LPO, FIC activity, HO* and O(2)(-) scavenging activities. Among them HEF (pH 4) has shown potent antioxidant activity. HEF (pH 4) was also found effective in protecting plasmid DNA and HAS protein oxidation induced by HO*. Pre-treatment of HEF (pH 4) at 75 and 150 mg/kg body weight for 6 days caused a significant increase in the levels of CAT and SOD and decrease in the level of MDA content in liver, lungs, kidneys and blood when compared to CCl(4)-intoxicated rats. Eventually, the extracts were also screened for anti-QS activity. Of these extracts two showed QS inhibition: HEF (pH 4) and HCE. The results obtained strongly indicate that green pod of A. nilotica are important source of natural antioxidants.

Determination of antimicrobial resistance and virulence gene signatures in surface water isolates of Escherichia coli.

AIMS: To determine the occurrence of Escherichia coli harbouring virulence markers of shiga- or entero-toxins and resistance to antimicrobials in surface waters. METHODS AND RESULTS: Surface water samples were collected at six locations of the river Gomti. E. coli isolates (n = 90) were characterized for their pathogenic potential using polymerase chain reaction to detect virulence genes as well as their sensitivity to antimicrobial agents using disc diffusion methods. In this study, 57.8% of E. coli isolates exhibited resistance to three or more antimicrobial agents. Sensitivity to cephotaxime, gentamicin and norfloxacin was observed in 7.8%, 48.9% and 77.8% of isolates, respectively. Both stx1 and stx2 genes were present in 15.6% of isolates while remaining isolates had either stx1 (17.8%) or stx2 (6.7%). The stx1 gene (33.3%) was more prevalent than stx2 (22.2%). The results indicate that the LT1 and ST1 genes were positive in 21.2% of isolates. CONCLUSIONS: The presence of multi-drug resistance and virulence genes in E. coli isolated from surface water being used for domestic and recreational purposes may result in waterborne outbreaks. SIGNIFICANCE AND IMPACT OF THE STUDY: The data will be useful in monitoring surface waters for forecasting and management of waterborne outbreaks.

Oxidant-antioxidant imbalance in the erythrocytes of sporadic amyotrophic lateral sclerosis patients correlates with the progression of disease.

Free radicals are implicated in numerous disease processes including motor neuron degeneration (MND). Antioxidant defense enzymes: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSHPx), glutathione reductase (GR) and glucose-6-phosphate dehydrogenase (G-6-PDH) in the erythrocytes are capable of detoxifying reactive oxygen species produced endogenously or exogenously. In the present study, the extent of lipid peroxidation (LPO) and antioxidant defenses were evaluated in the erythrocytes of 20 sporadic amyotrophic lateral sclerosis (ALS) patients and 20 controls. We observed that lipid peroxidation in the erythrocytes of amyotrophic lateral sclerosis patients significantly increased with respect to controls (P<0.001). On the other hand, catalase activity was found to be significantly lower (P<0.001). The activities of glucose-6-phosphate dehydrogenase, glutathione reductase and glutathione levels were also found to be significantly reduced in ALS patients compared to healthy subjects (P<0.001, P<0.01 and P<0.01, respectively). It was further observed that lipid peroxidation started to increase and catalase, glutathione reductase, glucose-6-phosphate dehydrogenase enzyme activities and glutathione levels started to decrease as amyotrophic lateral sclerosis progressed from 6 to 24 months, suggesting a correlation between these parameters and duration of amyotrophic lateral sclerosis. This study confirms the involvement of oxidative stress during the progression of amyotrophic lateral sclerosis and the need to develop specific peripheral biomarkers.

Frequency of infection by hepatitis B virus and its surface mutants in a northern Indian population.

INTRODUCTION: The reported prevalence of hepatitis B virus (HBV) infection in the Indian general population varies from 2% to 11%. Epidemiological studies conducted so far have selection biases, since these included populations of defined age group, gender, social class, high-risk group, etc. The present study was designed to look for the molecular epidemiology of HBV infection in the rural and urban general populations in India. METHODS: Sera obtained from healthy volunteers during college and social service camps from parts of northern India were tested for HBsAg and anti-HBc using enzyme immunoassays and for HBV DNA using polymerase chain reaction and Southern blot hybridization. The amplification products were cloned and sequenced, and nucleotide and deduced amino acid sequences of the surface and polymerase genes were analyzed for mutations. RESULTS: Of the 730 subjects (rural 543, urban 187), 15 (2.1%) tested positive for HBsAg and 143 (19.5%) for anti-HBc; 10 were positive for both. The overall HBV exposure rate in the population was 20.3% (148/730). The HBsAg carrier rate was similar in the urban and rural populations (1.5% and 2.3%; p=ns), and anti-HBc positivity was lower in the urban population (8.5% vs. 23.3%; p<0.01). History of parenteral interventions or blood transfusion was associated with markers of exposure to HBV (10.2% vs. 4.6%; p=0.01). Among the 220 representative samples tested for HBV DNA, 14 (6.4%) were positive; of these, only four were positive for HBsAg or anti-HBc. Sequencing of a 388-nt segment of the S-gene from three individuals (two adw and one ayw subtype) revealed four mutations. Two and three of these led to amino acid changes in the HBV surface and polymerase genes, respectively; alterations in known cytotoxic T cell epitopes of HBV surface and polymerase proteins were observed in one individual each. None had the G587A mutation, which is known to be associated with loss of the 'a' determinant of HBsAg. CONCLUSION: Our study shows a high frequency of exposure to HBV infection in the Indian general population; a proportion of HBV infected persons were detectable only by molecular methods. The positivity rate was higher in the rural population.

High frequency of hepatitis B virus infection in patients with beta-thalassemia receiving multiple transfusions.

BACKGROUND AND OBJECTIVES: Hepatitis B virus (HBV) may occasionally be transmitted through transfusion of blood units that are hepatitis B surface antigen (HBsAg) negative but HBV DNA positive. Children with beta-thalassemia are particularly susceptible to HBV because they receive multiple blood transfusions. These children have high infection rates despite vaccination against HBV. Post-vaccination infections may be a result of viruses harbouring surface (S)-gene mutations (e.g. G587A) in a region critical for reactivity to antibody to hepatitis B surface antigen (anti-HBs). The true prevalence of HBV in individuals with beta-thalassemia has not been studied previously. PATIENTS AND METHODS: Seventy patients with beta-thalassemia (median age 6 years; range 8 months to 22 years; 49 male), who had received seven to 623 (median 61) units of blood each and three doses (10/20 micro g) of HBV vaccine (Engerix B) before presentation to us, were included in the study; 50 of the 70 patients had received transfusions prior to vaccination. Enzyme-linked immunoassay for serological markers [HBsAg, antibody to hepatitis B core antigen (anti-HBc) and quantitative anti-HBs] and polymerase chain reaction (PCR) followed by Southern hybridization for molecular detection of hepatitis B, was performed on all samples. The PCR-amplified product was cloned, sequenced and the nucleotide and deduced amino acid sequences for the HBV S and polymerase (P) genes were analysed for mutations. RESULTS: Four of 70 (5.7%) individuals with beta-thalassemia were HBsAg positive and 14 (20%) were anti-HBc positive. The prevalence of serological markers increased with number of transfusions (P < 0.01). Of 70 patients, 53 (75.7%) had an anti-HBs titre of > 10 IU/l following vaccination and 17 (24.3%) were non-responders (< 10 IU/l); 22 (31.4%) of the 70 were DNA positive. The frequency of HBV infection in beta-thalassemia was similar in vaccine responders and non-responders. The virus was of subtype ayw (genotype D) in the five DNA-positive samples in which a 388-nucleotide region of the S gene was sequenced. Mutations occurred at 13 positions in the S gene and at 10 positions in the P gene. Hydrophobicity plots revealed differences in amino acid regions 117-165 and 195-211. Some of these amino acid substitutions coincided with the putative cytotoxic T-lymphocyte epitopes of both S and P proteins. CONCLUSIONS: A high frequency of HBV infection was seen using molecular methods in thalassemic patients. The frequency of infection was similar in vaccine responders and non-responders. A number of mutations were observed in the S gene, which could have implications for viral replication as well as virus-host cell interaction.

Mode of action of acibenzolar-S-methyl against sheath blight of rice, caused by Rhizoctonia solani Kühn.

The mode of action of acibenzolar-S-methyl (BTH) was investigated against sheath blight of rice and its pathogen, Rhizoctonia solani. BTH exhibited limited fungitoxicity against R solani, in the form of reduced mycelial growth, hyphal browning and sclerotia formation. Parasite fitness of mycelia and sclerotia formed on BTH-amended media was also reduced. When applied as soil drench or foliar spray, BTH inhibited both disease development on inoculated sheaths and its spread to the younger sheaths. The degree of protection against sheath blight increased with increase in duration between BTH application and inoculation. The curative effect of BTH was poor. When applied through roots a protective effect of BTH was visible even with only a 1-h interval between application and inoculation. However, in the case of foliar application, protective effect was recorded only when the gap between application and inoculation was 24 h. BTH reduced the frequency of penetration by R solani, colonization of host tissue and spread of the hyphae from primary lesions to form secondary lesions. BTH induced swelling of hyphal tips on the sheath surface, formation of papillae, browning of penetrated epidermal cells and degeneration of intra-cellular hyphae colonizing epidermal and mesophyll cells. Therefore, the protective effect of BTH against sheath blight was due to combination of its host defence-inducing activity and its adverse effect on growth and vigor (parasite fitness) of the pathogen.

Uptake and translocation of carpropamid in rice (Oryza sativa L).

Translocation of the antiblast compound, carpropamid, was investigated in rice using [14C]carpropamid. When applied to the seed, carpropamid was not only readily absorbed but was translocated to different parts of the seedlings emerging from treated seeds. A substantial portion of fungicide appeared to be exuded onto the leaf surface. In 21-day-old plants grown from [14C]carpropamid-treated seeds, 27.2% of the radioactivity isolated from leaves was present on the surface of lamina. This exuded fraction is probably responsible for its action as a fungal anti-penetrant compound. Following 30-min root dipping of 14-day-old seedlings, carpropamid was rapidly absorbed and translocated throughout the seedling. Its intra-laminar distribution was uniform as determined by autoradiography. Only a small fraction (< 2%) of fungicide applied to the foliage was translocated beyond the site of application within the treated leaf. Translocation was primarily apoplastic. Approximately 54% of the radioactivity recovered from leaves was in the form of carpropamid. At least seven radiolabelled metabolic products were observed by TLC. Only 8.3% of radioactivity applied through the seeds could be recovered from 21-day-old seedlings.

Artemisinin, an endoperoxide antimalarial, disrupts the hemoglobin catabolism and heme detoxification systems in malarial parasite.

Endoperoxide antimalarials based on the ancient Chinese drug Qinghaosu (artemisinin) are currently our major hope in the fight against drug-resistant malaria. Rational drug design based on artemisinin and its analogues is slow as the mechanism of action of these antimalarials is not clear. Here we report that these drugs, at least in part, exert their effect by interfering with the plasmodial hemoglobin catabolic pathway and inhibition of heme polymerization. In an in vitro experiment we observed inhibition of digestive vacuole proteolytic activity of malarial parasite by artemisinin. These observations were further confirmed by ex vivo experiments showing accumulation of hemoglobin in the parasites treated with artemisinin, suggesting inhibition of hemoglobin degradation. We found artemisinin to be a potent inhibitor of heme polymerization activity mediated by Plasmodium yoelii lysates as well as Plasmodium falciparum histidine-rich protein II. Interaction of artemisinin with the purified malarial hemozoin in vitro resulted in the concentration-dependent breakdown of the malaria pigment. Our results presented here may explain the selective and rapid toxicity of these drugs on mature, hemozoin-containing, stages of malarial parasite. Since artemisinin and its analogues appear to have similar molecular targets as chloroquine despite having different structures, they can potentially bypass the quinoline resistance machinery of the malarial parasite, which causes sublethal accumulation of these drugs in resistant strains.

Effect of protein malnutrition on sex organs of metanil yellow exposed male rats.

Oral administration of metanil yellow (MY) at 3.0% (w/w) dose level to adult male albino rats maintained on low protein (LP) diet for 30 days resulted in a greater decrease in absolute and relative weights of testes than in those rats maintained on a normal protein (NP) diet. A marked decrease in the activities of lactate dehydrogenase and hyaluronidase and content of lactic acid in LP + MY fed animals suggested that low protein diet enhanced the vulnerability of germ cells towards metanil yellow. The lack of significant changes in the cholesterol content of testis, the fructose content of the coagulating glands and the dorso-lateral prostate, the activities of alkaline phosphatase in the seminal vesicle, and acid phosphatase in ventral prostate of the MY treated animals suggested that their androgenic status were not affected.

Gastric M1 mucin, an early oncofetal marker of colon carcinogenesis, is encoded by the MUC5AC gene.

Gastric M1 mucin and the MUC5AC gene show a similar oncofetal expression in the colon. Our aim was to determine whether M1 mucin is the product of the MUC5AC gene. A recombinant baculovirus encoding the C-terminal portion of the MUC5AC gene as a fusion protein was isolated and the immunoreactivity of the recombinant mucin (rM) toward M1 antibodies studied. Chicken antibodies also were raised against purified rM. Besides its reactivity with L56/C, a serum recognizing the bacterially expressed MUC5AC gene product, rM was endowed with M1 immunoreactivity: (i) rM-expressing cells were stained specifically with anti-M1 serum and with the monoclonal antibody (MAb) 21M1, defining the M1-f epitope; (ii) both L56/C and anti-M1 antibodies recognized the same bands in immunoblots of rM-containing cell extracts; (iii) the 21M1 antibody reacted with rM in an immunoradiometric assay. Among the 7 M1 epitopes, M1-f was the only one encoded by the 3' portion of the MUC5AC gene. It was the only epitope detected in a native mucin M1-derived 170 kDa bromelain proteolytic fragment. Furthermore, the staining patterns of human tissues obtained with either anti-rM chicken antibodies or anti-M1 antibodies were identical. We conclude that M1 immunoreactivity is encoded at least in part by the MUC5AC gene.

Synthetic peptides corresponding to a repetitive sequence of malarial histidine rich protein bind haem and inhibit haemozoin formation in vitro.

Synthetic peptides containing a repetitive hexapeptide sequence (Ala-His-His-Ala-Ala-Asp) of malarial histidine-rich protein II were evaluated for binding with haem in vitro. The pattern of haem binding suggested that each repeat unit of this sequence provides one binding site for haem. Chloroquine inhibited the haem-peptide complex formation with preferential formation of a haem chloroquine complex. In vitro studies on haem polymerisation showed that none of the peptides could initiate haemozoin formation. However, they could inhibit haemozoin formation promoted by a malarial parasite extract, possibly by competitively binding free haem. These results indicate this hexapeptide sequence represents the haem binding site of the malarial histidine-rich protein and possibly the site of nucleation for haem polymerisation.

P-aminodiphenylamine induced biochemical changes in sex organs of male albino rats.

Intraperitoneal administration of p-aminodiphenylamine (p-ADPA), an aromatic amine of wide industrial applications, / 42.5 mg/kg body weight for 180 days significantly decreased the activities of testicular lactate dehydrogenase and hyaluronidase and lactic acid content indicating arrest of spermatogenesis. Patchy necrosis of the testis was confirmed histopathologically. No change in testicular cholesterol, fructose content of coagulating glands and dorso-lateral prostate and activities of alkaline phosphatase in seminal vesicle and acid phosphatase in ventral prostate support normal androgenic status.