Effect of unsaponifiable fraction of seeds of Hygrophila spinosa T. Ander on testosterone production of rat Leydig cells in vitro

Objective of the study: Seeds of Hygrophila spinosa (HS) T. Ander (Acanthaceae) are traditionally used as aphrodisiac and spermatogenic in Indian System of medicine. Preliminary phytochemical screening of plant revealed the presence of triterpenoids and sterols in seeds. The study was planned to assess the effect of unsaponifiable fraction prepared from seeds of HS on isolated rat Leydig cells for testosterone (T) production using in vitro method.Materials and methods: Leydig cells were isolated from wistar rats, aseptically, in vitro by collagenase cell dispersion method. Cells (2 X 106 cells/ml) were then incubated with unsaponifiable fraction of HS (10, 100 and 1000 µg/ml dose levels in triplicate) in incubator at 37°C under atmosphere of 95% CO2 condition for 3 hours in aseptic condition. Dehydroepiandrosterone (DHEA) was used as positive control in the study. The amount of T secreted in culture media was estimated using HPTLC. Benzene: Ethyl acetate (5:5% v/v) was employed as mobile phase and silica gel G F254 aluminum coated TLC plate as stationary phase.Results: The results indicated dose dependent increase in testosterone concentration in test groups. Isolated rat Leydig cells treated with the test fraction showed increased amount of testosterone present in culture media as compared to that of control.Conclusion: Unsaponifiable fraction prepared from seeds of HS showed ability to enhance biosynthesis of T in Isolated rat leydig cells. In vitro studies showed that the fraction might act locally in testis on Leydig cells and stimulated testosterone synthesis

SIMULTANEOUS ESTIMATION OF CURCUMINOIDS, PIPERINE, AND GALLIC ACID IN AN AYURVEDIC FORMULATION BY VALIDATED HIGH-PERFORMANCE THIN LAYER CHROMATOGRAPHIC METHOD

ABSTRACTObjective: The present study was proposed to quantitatively estimate the amount of three marker compounds; curcuminoids, piperine, and gallicacid in a multicomponent ayurvedic formulation using high-performance thin layer chromatographic (HPTLC) method for routine analytical work.Methods: TLC separation was performed on silica gel 60 F plates using toluene:ethyl acetate:formic acid:methanol (5.6:2.2:1.2:1.0 v/v/v/v) asmobile phase. Plate was developed by to a distance of 90 mm at ambient room temperature with 20 minutes saturation time. Densitometric analysiswas performed at 327 nm. Method was validated as per International Conference on Harmonization Q2 (R1) guideline also.254Results: Piperine, curcuminoids, and gallic acid were separated on TLC at retention factor values of 0.71, 0.61, and 0.29, respectively. The describedmethod was linear over the range of 300-700 ng/spot, 100-300 ng/spot, and 250-550 ng/spot, respectively, for curcuminoids, piperine, and gallicacid. The accuracy of the method was assessed by recovery studies and was found to be 101.71%, 99.67%, and 99.59% for curcuminoids, piperine,and gallic acid, respectively. The amount of curcuminoids, piperine, and gallic acid in the ayurvedic formulation was found to be 3.99% w/w, 1.9%w/w, and 0.8% w/w, respectively, when analyzed quantitatively by developed validated HPTLC method.Conclusion: The method can be used as a tool for quality control of herbal formulation.Keywords: Curcuminoids, Piperine, Gallic acid, High-performance thin layer chromatographic

ESTIMATION OF GUGGULSTERONE-Z IN GOKSHURADI GUGGULU USING REVERSED-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

Objective: A study was aimed to estimate guggulsterone-Z (GZ) in Gokshuradi Guggulu (GG).Methods: An analytical method was developed and validated using Waters Alliance high-performance liquid chromatography system (Empower software), equipped with photodiode array detector. Separation was achieved using Phenomenex, C-18 (250 mm×4.6 mm, 5 μ) column. Mobile phase consisted of acetonitrile:water (70:30,v/v). Flow rate was set to 1 ml/min and detection was performed at 251 nm.Results and Discussion: Validation parameters such as linearity, precision, accuracy, limit of detection, limit of quantification, and robustness were performed. Amount of GZ was estimated using linearity equation.Conclusion: GG was found to contain 0.815±0.03 g% w/w GZ. Validated method may be used as one of the parameters to standardize the formulation

Central venous catheter adverse events are not associated with crowding indicators

OBJECTIVE: Crowding in the emergency department (ED) impacts a number of important quality and safety metrics. We studied ED crowding measures associated with adverse events (AE) resulting from central venous catheters (CVC) inserted in the ED, as well as the relationship between crowding and the frequency of CVC insertions in an ED cohort admitted to the intensive care unit (ICU). METHODS: We conducted a retrospective observational study from 2008-2010 in an academic tertiary care center. Participants undergoing CVC in the ED or admitted to an ICU were categorized by quartile based on the following: National Emergency Department Overcrowding Scale (NEDOCS); waiting room patients (WR); ED patients awaiting inpatient beds (boarders); and ED occupancy (EDO). Main outcomes were the occurrence of an AE during CVC insertion in the ED, and deferred procedures assessed by frequency of CVC insertions in ED patients admitted to the ICU. RESULTS: Of 2,284 ED patients who had a CVC inserted, 293 (13%) suffered an AE. There was no association between AEs from ED CVCs and crowding scales when comparing the highest crowding level or quartile to all other quartiles: NEDOCS (dangerous crowding [13.1%] vs other levels [13.0%], P = 0.98); number of WR patients (14.0% vs 12.7%, P = 0.81); EDO (13.0% vs 12.9%, P = 0.99); and number of boarding patients (12.0% vs 13.3%), P = 0.21). In a cohort of ED patients admitted to the ICU, there was no association between CVC placement rates in the ED and crowding scales comparing the highest vs all other quartiles: NEDOCS (dangerous crowding 16% vs all others 16%, P = 0.97); WR patients (16% vs 16%, P = 0.82), EDO (15% vs. 17%, P = 0.15); and number of boarding patients (17% vs 16%, P = 0.08). CONCLUSION: In a large, academic tertiary-care center, frequency of CVC insertion in the ED and related AEs were not associated with measures of crowding. These findings add to the evidence that the negative effects of crowding, which impact all ED patients and measures of ED performance, are less likely to impair the delivery of prioritized time-critical interventions

TRANSITION METAL COMPLEXES OF ARYL AZOPYARAZOLE: SYNTHESIS, CHARACTERIZATION AND MICROBICIDAL ACTIVITIES

ABSTRACT Reaction between 4-nitro phenyl acid hydrazide with 4-[N'-(1-Ethoxycarbonyl-2-oxo-propylidene)-hydrazino]-2-hydroxy-benzoic acid in ethanol furnishes 1-[4-nitro-benzoyl]-3-methyl-4-(4-carboxy-3-hydroxy-phen-4-ylhydrazono)-2-pyrazoline-5-ones (NtH-ASA). The transition metal complexes of Cu 2+ , Co 2+ ,Ni 2+ , Mn 2+ and Zn 2+ of NtH-ASA have been prepared and characterized by elemental analyses, spectral studies, magnetic moment determination, molar conductivity measurement and microbicidal activity

ISOLATION AND CHEMICAL CHARACTERIZATION OF BIOACTIVE ALKALOID FROM ARGYREIA SPECIOSA LINN. HAVING ACTION ON ISOLATED RAT LEYDIG CELLS

Objective: Present study was aimed to isolate and chemically characterize bioactive constituent from alkaloid enriched fraction, prepared from roots of Argyreia speciosa Linn. Materials and methods: Literature review revealed presence of ergoline type of alkaloids in roots. Alkaloidal fraction was prepared and screened for its action on testosterone biosynthesis, in- vitro, using isolated rat leydig cells. Dehydroepiandosterone was used as positive control. This bioactive fraction was subjected to open column chromatography followed by flash chromatography, to isolate constituent. One compound (A1) was isolated from the fraction and its purity was ascertained using TLC and HPLC studies. A1 was chemically characterized by IR, Mass and 1H-NMR studies, to elucidate probable chemical structure. A1 was screened for action on testosterone synthesis too, using isolated rat Leydig cells model. The fraction was standardized with respect to amount of A1 present.   Results: Alkaloidal fraction (1000 µg/ml) incubated Leydig cells showed nearly, 22 fold increase in testosterone content as compared to untreated cells. The studies also yielded increase in testosterone content, in cells treated with test fractions and as observed in case of positive control. TLC studies indicated that A1 might possess ergoline moiety in the structure. IR, Mass and 1H NMR spectral studies suggested that A1 might be N-methyl ergometrine. This was the first report included isolation and chemical characterization of N-methyl ergometrine from Argyreia speciosa. A1 (1000 µg/ml) was found to stimulate testosterone content, by 14.62 fold, in culture media of Leydig cells after incubation. Conclusion: The results of in vitro studies, confirmed that the standardized alkaloid fraction as well as A1 had ability to stimulate Leydig cells to secrete testosterone. A1 might be N-methyl ergometrine and being ergometrine derivative it might act through oxytocine receptors expressed on the Leydig cells and stimulates testosterone synthesis

Prospective evaluation of risk factors for mortality in patients of Fournier′s gangrene: A single center experience

Introduction: Fournier′s gangrene is an aggressive disease with high morbidity and mortality. The aim of this study was to assess risk factors associated with mortality among patients of Fournier′s gangrene. Materials and Methods: Between May 2011 and September 2012, all patients of Fournier′s gangrene treated at our center were included in the study. All patients underwent emergency surgical debridement and received broad spectrum intravenous antibiotics. Their baseline characteristics, treatment, and follow-up data were recorded and analyzed. Results: A total of 30 patients were included in the study. Of these, six patients (20%) died during the treatment. Age >55 years, total leukocyte count >15000 cumm, extent of the area involved, septic shock at admission, visual analog scale (VAS) >7 at admission, and Fournier gangrene severity index (FGSI) score >8 at admission were significantly associated with increased mortality. Conclusion : In patients of Fournier′s gangrene, increased age, total leukocyte count, extent of the area involved, septic shock at admission, VAS score, and FGSI score at admission have a significant association with mortality