Study on the Epidemiology and Clinical Picture of Human Parainfluenza Virus in Chennai. Standardization of Rapid Diagnostic Tool and to Study the Effect of Hemagglutinin Neuraminidase Inhibitors on the Isolates

Human parainfluenza viruses are a group of viruses that cause different types of respiratory infections and are most common in children and infants. Throat and nasal swabs were collected from symptomatic patients in Chennai within three days onset of illness were determined the prevalence of HPIV by Multiplex reverse transcription PCR. Epidemiology of specific viral etiology in patients was observed throughout the years. The age wise distribution of HPIV cases were analyzed and divided into 0-10, 11-20, 21-30, 31-40, 41-50 and above 50. The prevalence in different age groups was statistically analyzed by standard error mean. Their positivity was observed in all the years during monsoon months of August to September and post monsoon months of November to February. Among the four serotypes HPIV type 3 is highly predominant in all the years (2011-2014). HPIV-2 positivity were occurred rarely in 2014. This study validates the prevalence of HPIV infection in Chennai and indicates the circulating serotypes and HPIV strains. The PCR products were sequenced and submitted to genbank and assigned the accession number. Different sequences were retrieved from NCBI and aligned as FASTA format. Mutations were identified by multiple sequence alignment of HPIV by ClustalW tool. Amino acid alterations were identified in HPIV-3 (HN gene) at residue 295 which Histidine replaced by Tyrosine and at 297 which Serine replaced by Glycine. Another mutations were identified in HPIV-2 (N gene) at residue 138 which Histidine replaced by Tyrosine and at 140th residue identified amino acid alteration which Histidine replaced by Glutamine. The phylogenetic analysis were identified the homology of Chennai strains with other strains. HPIV type 3 (HN) strain was clustered with Fukuoka /2009, Nagasaki 2009 and Wash 64979. HPIV type 3 (NP) strain was grouped with Switzerland/2013, US/2000, and South Africa/2000. HPIV type 2 was compared with Greer strain and HPIV2/V94. All clinical samples were cultured in the LLC-MK2, A549 and MDCK cell lines. Then specificity and sensitivity of the cell passages were characterized for clinical isolates. The number of positive cases were highly significant in the year 2011 followed by 2013. Out of 931 samples, 38 were isolated by LLC-MK2, 15 samples identified CPE in A549 and only 5 samples were grew in MDCK cells. Among the three cell lines LLC-MK2 was highly predominant for the isolation of HPIV. Positive percentage remained very small in MDCK cell line thus for further confirmation were not studied, whereas other two cell lines LLC-MK2 and A549 performed further confirmation. The isolated Human parainfluenza virus type 2 were more sensitive in the early passages of 8 and 9 at day five for LLC-MK2, highly compatible in the 5th passage at day 7 for A549 and 8th passage at day 8 for MDCK passage at day 8 for rest of the passages were less sensitive and specificity. HPIV-3 was more accustomed in the 9th passage at day 5 for LLC-MK2. The virus isolated samples were performed by hemadsorption assay (HAD) was aimed at confirmation of cytopathic effect were identified in LLC-MK2, A549 cell lines. Among the two cell lines LLC-MK2 was highly predominant to detect erythrocytes adhered on the monolayers and followed A549 cell lines. Cells infected with HPIV with C.Perfringens treatment by HAD assay to enhance erythrocyte binding for HPIV-2 (82%) and HPIV-3 (90%) in 24 well plate. Virus isolated samples were confirmed by plaque assay. Plaque was observed after 8-10 days of incubation. Interactions of receptors between HPIV-2 and 3 with m.o.i. in LLC-MK2 appeared as fusion were not blocked in HPIV-2 whereas HPIV-3 achieved fusion were blocked syncytium was not formed. Similar outcomes remained in A549 cells. Further the study evaluated the neuraminidase enzyme activity of HPIV. 4-MU concentrations and used determined the percentage of substrate expended during the reaction. The signal to background ratio were determined as the fluorescence intensities restrained after 20 minutes. The substrate concentrations were used at 25 μM. Various concentration of HPIV- 2 and 3 (m.o.i.) with bacterial neuraminidase were performed by neuraminidase activity which are statistically significant. Cytotoxicity of 4-GU-DANA at the concentrations less than 400 μM in A549 and greater than 641 μM in LLC-MK2 by MTT assay. Cytotoxicity of Ribavirin at the concentrations less than 405 μM in A549 and greater than 476 μM in LLC-MK2 by MTT assay. Cytotoxic percentage of Ribavirin were appears as high when compared with HN inhibitor. Cytotoxicity of glycyrrhizic acid form Licorice at concentration 31 μM in LLC-MK2 and 45 μM in A549 were identified. Indicates that MTT obtained better results compared with other dyes. Antiviral activity of neuraminidase inhibitor (4-GU-DANA) against HPIV by Hemadsorption inhibition assay was performed and ability to interfere with receptor interaction of HPIV-2 and 3 blocks hemadsorption activity at 600 μM seemed as 77% and 78% respectively. 4-GU-DANA inhibits receptor binding for HPIV-2 and 3 at 500 µM (60%) inhibit plaque formation in LL-CMK2 and (67%) and (79%) inhibit plaque reduction in A549 cells. In neuraminidase inhibition assay, less concentrations which inhibit the HPIV-2 and 3. The IC50 concentration for HPIV-2 at 2.5 μM and HPIV-3 at 1.6 μM. 4-GU-DANA concentrations were obtained less deliberation for HPIV-3 when compared to HPIV-2. Antiviral activity of nucleoside inhibitor (Ribavirin) against HPIV type 2 and 3 by hemadsorption inhibition assay was performed to inhibit the adherence of erythrocytes to the monolayer of HPIV type 2 and 3 at 400 μM (77%) and (75%) individually. Ribavirin inhibits replication for HPIV-2 and 3 at 400 μM during pre and post adsorption period in LLC-MK2 and A549 cells. There was no significant reduction in plaque number due to existence of ribavirin during the adsorption period of 90 minutes. The plaque area was reduced by addition after the adsorption period. Further confirmed inhibition of HPIV-2 by molecular characterized, observed infected cells without drug showed band by molecular characteristics. RBV treated infected cells, band cannot be seen and indicating that RBV inhibited transcription of viral genome. Cytotoxicity of Glycyrrihic acid from Licorice generated at 31 μM for LLC-MK2 and 45 μM for A549 by MTT assay. Antiviral activity of Glycyrrihic acid against HPIV-2 and 3 by hemadsorption inhibition assay was achieved to inhibit the erythrocyte adherence to the monolayer of HPIV-2 and 3 at 100 μM inhibition percentage was occurred 90% and 95% respectively. Glycyrrihic acid compound inhibits plaque formation at 70 μM replication of viral growth percentage was 86% for LLC-MK2 and 87% for A549 cells. Further confirmed by neuraminidase inhibition assay were performed at less concentration for HPIV-2 and 3 at 1.5 μM and 1.2 μM respectively. Neuraminidase inhibitor (Zanamivir) and nucleoside inhibitor (Ribavirin) were performed for antiviral activity against HPIV-2 and 3. Among these inhibitors Ribavirin has highly preferable with less concentration when compared to HN inhibitor. Among these three compounds natural glycyrrhizic acid from Licorice root were performed and observed very minimum concentration to inhibit both serotype of Human parainfluenza virus-2 and 3. The following inhibitors namely Neuraminidase inhibitor (Zanamivir) and nucleoside inhibitors (Ribavirin) maximum concentration was used to inhibit the HPIV-2 and 3. Finally, the phytal compound glycyrrhizic acid from licorice showed comparatively high inhibition on the viral growth in invitro screening. The Zanamivir nucleoside analog was elucidated for the mechanism of antiviral activity. The HN receptor of HPIV was docked with ligand Zanamivir using Autodock programme. All the HN receptor was significantly docked by Zanamivir. The 1V2i receptor was prominently docked with (95%) high frequency and good dock score

Random sampling of an AC source: A tool to teach probabilistic observations

An undergraduate level experiment is described to demonstrate the role of probabilistic observations in physics. A capacitor and a DC voltmeter are used to randomly sample an AC voltage source. The resulting probability distribution is analyzed to extract information about the AC source. Different characteristic probability distributions arising from various AC waveforms are calculated and experimentally measured. The reconstruction of the AC waveform is demonstrated from the measured probability distribution under certain restricted circumstances. The results are also compared with a simulated data sample. We propose this as a pedagogical tool to teach probabilistic measurements and their manipulations.Comment: Revtex4 file, 10 pages with 8 figure

Estimation of Serum Albumin Levels in Chronic Generalised Periodontits, Localized Aggressive Periodontitis Patients and Compare it with Periodontally Healthy Individuals

BACKGROUND: Serum albumin is a negative acute phase protein, primarily synthesized by liver. Many factor affects the regulation of serum albumin levels. The primary ones involved are inflammation and malnutrition. Chronic disease which are associated with inflammation and the release of inflammatory cytokines such as interleukin -1, interleukin-6 and tumor necrosis factor α cause decrease in albumin. Periodontitis is a chronic inflammatory disease caused by bacterial infection of the supporting tissues around the teeth. A few studies have shown that serum albumin levels are reduced in patients with Generalised chronic Periodontitis. AIM OF THE STUDY: 1. To estimate and compare the serum albumin levels in chronic generalised and localized aggressive Periodontitis patients with that of Periodontally heal thy individuals. 2. To identify whether chronic generalised periodontitis and localized aggressive periodontitis have any effect on serum albumin levels compared to Periodontally healthy individuals. MATERIALS AND METHODS: Total number of 60 subjects were enrolled in this study with the age range of 18 -50 years. All subjects participating in the study were informed about the nature of the study and al l individuals signed in the written informed consent form. 60 subjects were divided into 3 groups. Group I- Periodontally healthy individual, Group II- Chronic generalised periodontitis, Group III - Localized aggressive periodontitis. Clinical parameters such as Gingiva l bleeding index (GI), Periodontal pocket depth (PPD), Clinical attachment level (CAL) and serum albumin level were evaluated between these three groups. RESULTS: Al l the clinical parameters shown to be increased significantly in patients with chronic generalised periodontitis (Group II) and localized aggressive periodontitis (Group III) when compared to periodontally heal thy individuals (Group I). There is a significant decrease in serum albumin level in patients with chronic generalised periodontitis (Group II) and localized aggressive periodontitis (Group III) when compared to periodontally healthy individuals (Group I). CONCLUSION: With the limitations of the present study it could be concluded that there is inverse relationship of serum albumin level and periodontal disease. So, serum albumin level can be used as a predictive marker for severity of periodontitis. Further randomized clinical trials and longitudinal evaluations in a larger population would be required to support the observation of the present study

First simulation study of trackless events in the INO-ICAL detector to probe the sensitivity to atmospheric neutrinos oscillation parameters

The proposed India-based Neutrino Observatory will host a 50 kton magnetized iron calorimeter (ICAL) with resistive plate chambers as its active detector element. Its primary focus is to study charged-current interactions of atmospheric muon neutrinos via the reconstruction of muons in the detector. We present the first study of the energy and direction reconstruction of the final state lepton and hadrons produced in charged current interactions of atmospheric electron neutrinos at ICAL and the sensitivity of these events to neutrino oscillation parameters $\theta_{23}$ and $\Delta m_{32}^2$. However, the signatures of these events are similar to those from neutral-current interactions and charged-current muon neutrino events in which the muon track is not reconstructed. On including the entire set of events that do not produce a muon track, we find that reasonably good sensitivity to $\theta_{23}$ is obtained, with a relative $1\sigma$ precision of 15% on the mixing parameter $\sin^2\theta_{23}$, which decreases to 21%, when systematic uncertainties are considered

QCD Working Group Report

This is the report of the QCD working group at WHEPP 6. Discussions and work on heavy ion collisions, polarised scattering, and collider phenomenology are reported.Comment: Report of the QCD group at WHEPP-6, Chennai, January 2000. 7 page

Structure and Reactivity of Halogenated GC PNA Base Pairs – A DFT Approach

The present study explored the structural and reactivity relationship of halogenated G-C PNA base pairs using density functional theory (DFT) calculations. The halogens such as F, Cl, and Br are substituted by replacing H atoms involved in H-bonds of the base pairs. All structures were optimized using the B3LYP/6-311++G** theory level, and positive frequencies confirmed their equilibrium states. To understand the structural variations of the considered halogenated systems, the bond distances of R─X, R─H, and X/H•••Y and the bond angles of R─X•••Y were analyzed. The obtained structural parameters and interaction energies are comparable with the previous theoretical reports. In addition, the interaction energies (Eint) and quantum molecular descriptors (QMD) are also calculated to understand the difference between halogenated PNA systems and their non-halogenated counterparts. In this study, the enhancement in the reactivity properties  of halogenated PNA systems has been demonstrated, which indicates their improved responsive characteristics in various chemical reactions. Based on the available results, the halogenated PNA systems, carefully considering their substitutional position, facilitate better accommodation for the triplex formation of dsDNA/dsRNA. Therefore, it is concluded that the improved reactivity properties of halogenated PNA base pairs would make them potential candidates for various biological applications

Ethyl 2-methyl-5-oxo-4-(3,4,5-trimeth­oxy­phen­yl)-1,4,5,6,7,8-hexa­hydro­quinoline-3-carboxyl­ate

In the mol­ecular structure of the title compound, C22H27NO6, the dihydro­pyridine ring adopts a flattened boat conformation while the cyclo­hexenone ring is in an envelope conformation. In the crystal, mol­ecules stack parallel to the crystallographic a axis linked by inter­molecular N—H⋯O and C—H⋯O hydrogen bonds

Extraction process optimization of flavonoid and in vitro amylase inhibitory effect of purified quercetin derivative from Amorphophallus paeoniifolius tubers

544-556Amorphophallus paeoniifolius (Elephant foot yam) is a prominent tuberous plant utilized across several parts of India to treat various ailments such as a tumour, haemorrhage, microbial infections, cough, bronchitis, diabetes, anaemia, and hepato-gastro and cardiovascular diseases. In this context, the present study aims to optimize the extraction process of the flavonoid and to study the in vitro amylase inhibitory effect of purified flavonoid moiety. The Shake flask method with different extraction solvents was adopted to quantify the flavonoid content. Central composite design (CCD) based response surface methodology (RSM) was formulated to optimize the extraction process. Three-dimensional preparative chromatography (3D PTLC) was executed to purify the flavonoid content and high-resolution liquid chromatography-mass spectrometry (HRLC-MS) was adopted to predict the structure. 3,5-dinitrosalicylic acid (DNS) based spectrophotometry method was used to determine the amylase inhibitory property. All the analyses were subjected to standard statistical tests. The developed model for the extraction optimization process was found to be near significant (P = 0.242) with temperature as a significant variable (P = 0.029), and a 107-fold increase (71.11±0.5 mg/g tissue) of flavonoid content was recorded. A strong yellow colour spot (flavonoid fraction) was eluted using 3D PTLC technique and the molecule was identified as quercetin derivative (m/z 447) by the direct MS method. Significant amylase inhibition (36.1±2.1%) recorded by purified quercetin derivative has documented the utilization of A. paeoniifolius tubers as classical traditional medicine

Identifying the Neutrino mass Ordering with INO and NOvA

The relatively large value of $\theta_{13}$ established recently by the Daya Bay reactor experiment opens the possibility to determine the neutrino mass ordering with experiments currently under construction. We investigate synergies between the NOvA long-baseline accelerator experiment with atmospheric neutrino data from the India-based Neutrino Observatory (INO). We identify the requirements on energy and direction reconstruction and detector mass for INO necessary for a significant sensitivity. If neutrino energy and direction reconstruction at the level of 10% and 10 degree can be achieved by INO a determination of the neutrino mass ordering seems possible around 2020.Comment: 18 pages, 8 figures, minor improvements and clarifications, new panel in fig. 7, version to appear in JHEP, typo in eq. 4 correcte

J/\psi production through resolved photon processes at e+ e- colliders

We consider J/psi photoproduction in e+ e- as well as linear photon colliders. We find that the process is dominated by the resolved photon channel. Both the once-resolved and twice-resolved cross-sections are sensitive to (different combinations of) the colour octet matrix elements. Hence, this may be a good testing ground for colour octet contributions in NRQCD. On the other hand, the once-resolved J/psi production cross-section, particularly in a linear photon collider, is sensitive to the gluon content of the photon. Hence these cross-sections can be used to determine the parton distribution functions, especially the gluon distribution, in a photon, if the colour octet matrix elements are known.Comment: Added a figure on parametrisation dependence of photonic parton densities and some reference