Water utilization in urban households: economic constraint, and environmental concern in Guwahati, Assam

by Manish Kumar, Omi Kumari, Arbind K. Patel and Pinky Tanej

Bioactivity guided fractionation of leaves extract of Nyctanthes arbor tristis (Harshringar) against P falciparum.

BACKGROUND: Nyctanthes arbor-tristis (Harshringar, Night Jasmine) has been traditionally used in Ayurveda, Unani and other systems of medicine in India. The juice of its leaves has been used by various tribal populations of India in treatment of fevers resembling malaria. AIM OF THE STUDY: This work reports the antiplasmodial activity guided fractionation of Harshringar leaves extract. METHODOLOGY: Crude ethanolic Harshringar leaves extract and its RPHPLC purified fractions were studied for antiplasmodial potency against 3D7 (CQ sensitive) and Dd2 (CQ resistant) strains of P.falciparum and subsequently subjected to bioassay guided fractionation using reverse phase chromatography to pursue the isolation of active fractions. PRINCIPAL FINDINGS: Harshringar crude leaves extract and some of its RPHPLC purified fractions exhibited promising antiplasmodial potency against 3D7 and Dd2 strains of P.falciparum. CONCLUSIONS: The present study has provided scientific validity to the traditional use of leaves extract of Harshringar against malaria leading to the conclusion that this plant holds promise with respect to antimalarial phytotherapy. This is the first scientific report of antiplasmodial activity of RPHPLC fractions of Harshringar leaves extract against P.falciparum strains

Difference Spectroscopic Titration and formation of β-hematin.

<p>Panels A and B show difference absorption spectra of heme binding to BNT1 and BNTM respectively. Panel C shows heme-binding curves of BNT1 and BNTM generated from the difference absorption spectra by plotting A₄₁₅ vs the moles of heme/mole of peptide template. Horizontal lines were drawn connecting the Y axis to the plateau region of the binding curves. At the point of intersection, vertical arrows were drawn to obtain the values of heme/peptide.Panel D shows comparative kinetics of heme binding to BNT1/BNTM (at heme: peptide molar ratio of 4) studied by difference spectroscopy. Data shows intensity of Soret peak as a function of time. Panel E shows comparative kinetics of β-hematin formation in presence of BNT1/BNTM.</p

Antiplasmodial potencies of Reverse phase chromatographic fractions of Harshringar leaves extract.

<p>Data shown are expressed as the mean of triplicates ± SD. Potencies for active fractions (IC₅₀<8 µg/mL) are shown in italics.</p>a<p>Fractions which were combined together,</p>b<p>Time boundaries given correspond to the vertical lines shown in each chromatogram of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051714#pone-0051714-g003" target="_blank">Fig. 3B</a>,</p>c<p>not done.</p><p>ACN: Acetonitrile, RPCC- Reverse phase glass column chromatography. CQ (100 nM, 1000 nM) were used as positive controls for <i>Pf</i>3D7 and <i>Pf</i>Dd2 respectively. For details of chromatographic separations, see Materials and methods and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051714#pone-0051714-g001" target="_blank">Figures 1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0051714#pone-0051714-g003" target="_blank">3</a>. Symbols used for varying antiplasmodial potency are *** (Promising Activity, IC₅₀: 5–15 µg/mL), ** (Moderate Activity, IC₅₀: >15–50 µg/mL), and * (Inactive, IC₅₀: >50 µg/mL).</p

UV-visible spectra of fractions of Harshringar leaves extract.

<p>(A) Spectra of 40%, 60%, 80%, 100% MF were recorded against methanol as blank. 110 µg/mL, 90 µg/mL, 180 µg/mL, 80 µg/mL of 40%, 60%, 80%, 100% MF respectively were used for spectral studies. (B) UV-visible spectra of pool of 60%–80% MF (40 µg/mL).</p

Immunonano-Lipocarrier-Mediated Liver Sinusoidal Endothelial Cell-Specific RUNX1 Inhibition Impedes Immune Cell Infiltration and Hepatic Inflammation in Murine Model of NASH

Background: Runt-related transcription factor (RUNX1) regulates inflammation in non-alcoholic steatohepatitis (NASH). Methods: We performed in vivo targeted silencing of the RUNX1 gene in liver sinusoidal endothelial cells (LSECs) by using vegfr3 antibody tagged immunonano-lipocarriers encapsulated RUNX1 siRNA (RUNX1 siRNA) in murine models of methionine choline deficient (MCD) diet-induced NASH. MCD mice given nanolipocarriers-encapsulated negative siRNA were vehicle, and mice with standard diet were controls. Results: Liver RUNX1 expression was increased in the LSECs of MCD mice in comparison to controls. RUNX1 protein expression was decreased by 40% in CD31-positive LSECs of RUNX1 siRNA mice in comparison to vehicle, resulting in the downregulation of adhesion molecules, ICAM1 expression, and VCAM1 expression in LSECs. There was a marked decrease in infiltrated T cells and myeloid cells along with reduced inflammatory cytokines in the liver of RUNX1 siRNA mice as compared to that observed in the vehicle. Conclusions: In vivo LSEC-specific silencing of RUNX1 using immunonano-lipocarriers encapsulated siRNA effectively reduces its expression of adhesion molecules, infiltrate on of immune cells in liver, and inflammation in NASH