Screening of Nepalese crude drugs traditionally used to treat hyperpigmentation: in vitro tyrosinase inhibition.

South-East Asian population is daily exposed to strong sunlight. As a result, the majority of population will have darker, ethnic skin. Moreover, many people suffer from dark spots, hyperpigmentation, which is considered to be a skin disorder and causes psychological disturbance. To treat dark spots, most of the population will still rely on traditionally used crude drugs, knowledge about which is transferred from generation to generation. Fifty-two crude drugs were selected based on the survey performed among local healers and beauticians of different ethnic origin. These crude drugs were screened for mushroom tyrosinase inhibitory activity, as tyrosinase inhibitors are becoming increasingly important as cosmetic and medicinal products, primarily to control hyperpigmentation. Among the tested crude drugs, methanolic extracts of Glycyrrhiza glabra, Morus alba, Syzygium aromaticum, Citrus aurantifolia, Cypreae moneta, Punica granatum and Citrus aurantium, at the final concentration of 50 microg mL(-1), showed mushroom tyrosinase inhibitory activity of 78.9%, 71.0%, 69.4%, 59.0%, 56.0%, 53.4 and 51.9%, respectively, with 91.4% inhibitory activity of kojic acid taken as positive control. To our knowledge, this is the first report that extracts of Cypreae moneta shell and Syzygium aromaticum flowering bud have tyrosinase inhibitory activity. These potent extracts were further evaluated at different concentration. The final concentration of the extracts in reaction mixtures was 50, 25 and 5 microg mL(-1) for the initial concentration of 1000, 500 and 100 microg mL(-1), respectively. They showed concentration-dependent inhibition of mushroom tyrosinase. Those extracts expressing relatively weak tyrosinase inhibitory activity may act through different inhibition pathway which is not based on tyrosinase activity. Further evaluation of the most potent tyrosinase inhibitors in in vivo conditions would be recommended.

An integrated treatment approach: a case report for dentinogenesis imperfecta type II.

Dentinogenesis imperfecta type II or hereditary opalscent dentin is one of the most common autosomal dominant anomaly of dentin that occurs in both sex affecting approximately 1:8000 persons. Clinically this disorder is characterized by variable blue gray to yellow brown teeth, with fracture of enamel and excessive wear. The treatment strategy is focused towards protecting teeth from further wear and tear and total oral rehabilitation of patient with paramount importance to aesthetics, obtaining an appropriate vertical dimension and providing soft tissue support which will help to return the facial profile to a more normal appearance. A multidisciplinary treatment planning is required for treatment of these individuals.

Retrieval of a separated file using Masserann technique: a case report.

The separated instrument particularly a broken file leads to metallic obstruction, in the root canal and blocks thorough cleaning and shaping procedure. When attempts of bypassing such a fragment go in vain, it should be retrieved by mechanical devices. Masserann Kit is one such device for orthograde removal of intracanal metallic obstructions. This clinical case demonstrates usage of Masserann technique in successful retrieval of a separated file which was tightly binding in the apical 3rd root canal dentin of maxillary left lateral incisor.

Profile of Patients Admitted in Maternal Intensive Care Unit at BPKIHS, a Tertiary Hospital in Eastern Nepal.

BACKGROUND: Maternal death is a tragic event. It can be reduced by prompt recognition of critical illness in pregnancy and earlier initiation of intensive care. The physiological changes of pregnancy and the presence of a fetus complicates the assessment and management of critically ill obstetric patients. The objective of this study was to analyse the basic contributing factors for maternal intensive care unit admission and the maternal outcome. METHODS: This was a prospective study conducted in the department of Obstetrics and Gynaecology at B.P. Koirala Institute of Health Sciences, a tertiary hospital in eastern Nepal, for one year duration from January-December 2012. Data like age, diagnosis at admission, intervention, indication for admission, duration of stay and outcome were analysed. RESULTS: One hundred and ninety two patients were admitted in one year. Among them 177 were obstetrics related admission and 15 were non obstetrics. Out of 177 patients, 21(11.8%) were antenatal, 123(69.4%) were postnatal and 33(18.6%) had early pregnancy complications. The mean age in years was 25.67±7.169. One hundred and seventy one patients (96.6%) were unbooked and only 6(3.3%) were booked. Among the postnatal patients, 83(67.4%) had delivered at BPKIHS, 24(19.5%) at other health centres and 16 (13%) at home. Antepartum eclampsia was the commonest diagnosis. Out of 192 patients, 148(78.12%) were improved, 24(12.5%) had expired, 15(7.8%) went against medical advice and 5(1.6%) were referred. The median duration of MICU stay in days (IQR) was 2(1-4). CONCLUSIONS: An intensive care unit often offers the opportunity to improve the patient care.

Inhibitory effect of tetrahydroswertianolin on tumor necrosis factor-alpha-dependent hepatic apoptosis in mice.

We investigated the effect of tetrahydroswertianolin (THS), a hepatoprotective agent from Swertia japonica, on tumor necrosis factor-alpha (TNF-alpha)-dependent hepatic apoptosis induced by D-galactosamine (D-GalN) (700 mg/kg, i.p.) and lipopolysaccharide (LPS) (10 microg/kg, i.p.) in mice. Apoptotic symptoms were observed at the initial stage of liver damage. By 5 hr after intoxication, hepatic DNA fragmentation had risen to 2123%, with the value in untreated mice set at 100%, without a significant elevation of serum alanine transaminase (ALT) activity. There was a parallel increase in hepatocytes undergoing chromatin condensation and apoptotic body formation. By 8 hr after intoxication, serum ALT activity had risen to 3707 U/L. Pretreatment with THS (50 mg/kg, p.o.) at 18 and 2 hr before intoxication significantly reduced DNA fragmentation to 821% of that in untreated mice and prevented the emergence of chromatin condensation and apoptotic body formation. A significant and dose-dependent reduction in serum ALT activity at 8 hr also was observed with THS pretreatment. These effects of THS were different from those observed from pretreatment with glycyrrhizin (GCR), which is a clinically used hepatoprotective agent with membrane-stabilizing activity. GCR pretreatment (100 mg/kg, p.o.) did not inhibit hepatic DNA fragmentation (1588% of untreated mice), although this compound significantly protected against serum ALT elevation (1463 U/L). These data suggest that an inhibitory effect on the progression of hepatic apoptosis prior to liver injury may be involved in the hepatoprotective mechanisms of THS, whereas it appears that GCR affects the processes after apoptosis. In a separate experiment, we found that the concentration of serum TNF-alpha rose to 2016 pg/mL at 1 hr after intoxication of mice with D-GalN and LPS, but this increase was suppressed by THS pretreatment (10, 50, or 200 mg/kg, p.o.) to 716, 454, or 406 pg/mL, respectively. Further study with a reverse transcriptase-polymerase chain reaction method showed that THS blocked TNF-alpha production at the transcriptional level. Because TNF-alpha is a critical mediator to elicit apoptosis in this model, the property of suppressing TNF-alpha production may be of prime importance for THS inhibition of hepatic apoptosis.

Calyxin H, Epicalyxin H, and Blepharocalyxins A and B, Novel Diarylheptanoids from the Seeds of Alpinia blepharocalyx

Four unprecedented diarylheptanoids-calyxin H (1) and epicalyxin H (2), possessing a diarylheptanoid unit and a chalcone moiety, and blepharocalyxins A (3) and B (4), possessing two diarylheptanoid units and a chalcone moiety-were isolated from the seeds of Alpiniablepharocalyx. The structures of 1-4, including absolute stereochemistry, were elucidated by spectroscopic means and after a consideration of their biogenesis.

Potent free radical scavenging activity of propol isolated from Brazilian propolis.

We evaluated free radical scavenging activity of the water, methanol and chloroform extracts of propolis in 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical and xanthine-xanthine oxidase (XOD) generated superoxide anion assay systems. The free radical scavenging activity guided fractionation and chemical analysis led to the isolation of a new compound, propol (3-[4-hydroxy-3-(3-oxo-but-1-enyl)-phenyl]-acrylic acid) from the water extract, which was more potent than most common antioxidants such as vitamin C and vitamin E (alpha-tocopherol) in these assay systems.

Induction of Labor Using Misoprostol With or Without Mifepristone in Intrauterine Death.

INTRODUCTION: Rapid expulsion of fetus in intrauterine fetal death is usually requested without any medical grounds for it. So an efficient, safe method for induction of labor is required. The objective of this study is to determine if pre-treatment with mifepristone followed by induction of labor with misoprostol in late intrauterine fetal death is more efficacious. METHODS: We conducted a randomized controlled trial in 100 patients in B.P.Koirala Institute of Health Sciences, Nepal from June 2011 to May 2013. Group A women received single oral dose of 200 mg mifepristone, followed by induction with vaginal misoprostol after 24 hours. Group B women were induced only with vaginal misoprostol. In each group, five doses of misoprostol was used four hourly. If first cycle was unsuccessful, after break of 12 hour, second course of misoprostol was started. The primary outcome was a measure of induction to delivery time and vaginal delivery within 24 hours. Secondary outcome was to measure need of oxytocin and complications. RESULTS: Maternal age, parity and period of gestation were comparable between groups. Number of misoprostol dose needed in group A was significantly less than group B. Mann Whitney U test showed, women in group A had significantly earlier onset of labor, however total induction to delivery interval was not significant. In group A, 85.7% delivered within 24 hours of first dose of misoprostol while in group B 70% delivered within 24 hours (p=0.07). More women in Group B required oxytocin. CONCLUSIONS: Pretreatment with mifepristone before induction of labor following late intrauterine fetal death is an effective and safe regimen. It appears to shorten the duration of induction to onset of labor.

Pandemic influenza A (H1N1) 2009 outbreak investigation in Nepal.

BACKGROUND: The World Health Organization, on 24 April 2009, announced a Public Health emergency of international concern caused by a new influenza virus Pandemic Influenza A 2009. The objective of this study was to analyze the basic epidemiology and distribution of Pandemic Influenza A 2009 in order to understand the course of Pandemic Influenza A 2009 in Nepal. METHODS: The analyses were based upon all confirmed and probable cases that consulted Avian Influenza Control Project and National Public Health Laboratory during 29 April 2009 to 21 September 2010. RESULTS: Out of total 739 suspected samples collected, Pandemic Influenza A 2009 was detected in 210 cases in different districts of Nepal. The majority of cases were from the urban settlement of Kathmandu valley, Chitwan and Kaski and among age group 11-30 years. The clinical attack rate for Influenza like illness (ILI) was 28.48%. There was no significant difference between the clinical presentation of ILI and confirmed cases of Pandemic Influenza A 2009. CONCLUSIONS: This study presented the investigation of outbreak that helped to inform the course of epidemic in affected population and therefore urge for public health interventions.

Microalbuminuria among type-2 diabetes mellitus patients in Pokhara, Nepal.

Microalbuminuria is considered to be an early stage of diabetic nephropathy as well as a marker of cardiovascular disease. The aim of this study was to see the prevalence of microalbuminuria in type 2 diabetic patients and assess its association with cardiovascular risk factors among them. A total of 143 type 2 diabetic patients with the mean age of 56.06 +/- 1.08 years were analysed. The prevalence of microalbuminuria and overt proteinuria was 45.5% and 11.2%, respectively. Prevalence of microalbuminuria in female was marginally higher than in male (p > 0.05). Subjects with microalbuminuria had significantly higher blood pressure (p < 0.001) and duration of diabetes (p < 0.05) compared with normoalbuminuric subjects. High density lipoprotein was found to be significantly lower (p < 0.05) in subjects with microalbuminuria whereas fasting blood sugar, triglyceride, total cholesterol and very low density lipoprotein were marginally higher in microalbuminuric than in normoalbuminuric subjects (p > 0.05). High prevalence of microalbuminuria in diabetic patients and its positive association with blood pressure and altered lipid profile suggests that screening for microalbuminuria is essential for intervention and prevent further complications like end stage renal disease and cardiovascular diseases.

Tonsillar microbial flora: a comparison of infected and non-infected tonsils.

OBJECTIVE: The main objective of this study was to find out the most common organism affecting the tonsils in recurrent tonsillitis. METHODOLOGY: This was a prospective study consisting of total number of 50 patients, conducted in the ENT department of Dhulikhel Hospital, KUTH, Dhulikhel, Nepal. The study period was one year and three months (March 2004 to April 2005). RESULTS: Total numbers of 50 patients were selected, of which 32 were of acute tonsillitis and 18 were taken as control. Among these males were 32 and females were 18. The age group was from 3 years to 64 years. Throat swabs of patients were taken by aseptic method and were sent to the laboratory immediately. Among the infected tonsils the most common organism was Streptococcus viridans followed by Streptococcus pneumoniae and Moraxella catarrhalis. The third most common was Beta haemolytic streptococci whereas Micrococcus was not isolated in any of the infected samples. Likewise in non-infected tonsils, the most common organism isolated was again Streptococcus viridans followed by Micrococcus and Diptheroids. In these specimen no Beta haemolytic streptococci was isolated from the control group. CONCLUSION: Thus, the study is able to put forward the fact that most common organism affecting the tonsils in infected as well as non- infected state is Streptococcus viridans. Where as Beta haemolytic streptococci was not found in non-infected tonsils, likewise Micrococcus was not found in infected tonsils.

Anatomical variation of maxillary sinus mimicking a periapical cyst: a case report.

Maxillary sinus can pose diagnostic dilemma radiographically because of its anatomical variation which can mimic a periapical pathosis. This case report deals with one such diagnostic problem, where a maxillary sinus was interpreted in an intraoral periapical radiograph as a periapical cyst. With the advances in imaging techniques, the use of an Ultrasound imaging together with application of Colour Power Doppler helped in revealing the contents of the radiolucent area and come up with an accurate diagnosis. Thus a thorough knowledge about the normal anatomy and its variations and proper diagnostic aids are essential in the diagnosis of periapical pathology.

Immunostimulating activity of Celosian, an antihepatotoxic polysaccharide isolated from Celosia argentea.

Celosian, an acidic polysaccharide from the seeds of Celosia argenteo (Amaranthaceae) was found to be a potent antihepatotoxic agent for chemical and immunological liver injury models in animals. The immunomodulating action of celosian was studied to clarify the preventive mechanism of celosian on liver injuries. Celosian induced tumor necrosis factor-alpha (TNF-alpha) production in mice. Celosian also induced the production of interleukin-1 beta (IL-1 beta) and nitric oxide (NO) in macrophage cell line J774.1 in a concentration-dependent manner (1 to 1000 micrograms/ml). Moreover, celosian induced IL-1 beta secretion in human mononuclear cells. In addition, celosian enhanced gamma interferon (IFN-gamma) production activity of concanavalin A (Con A) in mice spleen cells, though celosian alone did not significantly influence IFN-gamma production. These results indicate that celosian is an immunostimulating agent in addition to antihepatotoxic effects.

Five new C-methyl flavonoids, the potent aldose reductase inhibitors from Matteuccia orientalis Trev.

Five new compounds, matteuorien, matteuorienin, and matteuorienate A (7), B (8), and C (9) were isolated together with five known compounds from the MeOH extract of the rhizome of Matteuccia orientalis Trev. The structures of these compounds were determined by the use of spectroscopic methods including two dimensional (2D)-NMR experiments and chemical methods, except for the configuration at the C-3"' of matteuorienate A (7), B (8), and C (9). Among them, matteuorienate A, B, and C showed very strong aldose reductase inhibitory activity. A structure-activity relationship study showed that a carboxyl group played an important part in aldose reductase inhibitory activity in these three compounds.

Two new 2-arylbenzofuran derivatives from hypoglycemic activity-bearing fractions of Morus insignis.

Ethyl acetate- and n-butanol-soluble fractions of the leaves of Morus insignis showed a significant hypoglycemic activity on streptozotocin (STZ)-induced hyperglycemic rats. From these hypoglycemic activity-showing fractions, two new compounds, mulberrofuran U (2) and moracin M-3-O-beta-D-glucopyranoside (3) were isolated, along with six known compounds [beta-sitosterol, beta-sitosterol-3-O-beta-glucopyranoside, ursolic acid, moracin M (1), kaempferol-3-O-beta-glucopyranoside and quercetin-3-O-beta-glucopyranoside] and the structures of the new compounds were determined by spectroscopic and chemical methods.

Bellidifolin: a potent hypoglycemic agent in streptozotocin (STZ)-induced diabetic rats from Swertia japonica.

Hypoglycemic activity-guided fractionation led to the isolation of five known xanthones and two triterpenoids from the ethyl acetate soluble fraction of Swertia japonica and their identification was based on spectroscopic methods. One of the triterpenes, thysanolactone, was first isolated from this plant. Among the xanthones, bellidifolin showed a potent and dose-dependent hypoglycemic activity in STZ-induced diabetic rats both in i.p. and p.o. administration. A comparative hypoglycemic activity of the other three xanthones together with bellidifolin was also studied.

Matteuorienate A and B, two new and potent aldose reductase inhibitors from Matteuccia orientalis (Hook.) Trev.

Matteuorienate A and B, two new C-methyl flavanone derivatives, were isolated from the Matteuccia orientalis (Hook.) Trev. (Aspidiaceae), and their structures were determined by the use of spectroscopic methods including 2D-NMR (1H-1H COSY, 1H-13C COSY and 1H-13C long-range COSY) experiments and chemical methods. Both the compounds were found to be very strong inhibitors of aldose reductase among the natural products.

Aldose reductase inhibitory constituents of the root of Salvia miltiorhiza Bunge.

The constituents of the MeOH extract of Salvia miltiorhiza BUNGE, which showed strong aldose reductase (AR) inhibitory activity, were examined, and two new abietane-type diterpenoids, danshenol A (1) and danshenol B (2), were isolated together with six known ones: dihydrotanshimme I (3), cryptotanshinone (4), tanshinone I (5), tanshinone IIA (6), (-)-danshexinkun A (7), and sugiol (8). Among them, 4, 5, and 8 were weak AR inhibitors with IC50 from 4.80 to > 10.0 microM, while 1, 2, 3, 6, and 7 were strong inhibitors (IC50 from 0.10 to 1.75 microM). Danshenol A (1), the strongest inhibitor, had IC50 of 0.10 microM which is comparable to that of epalrestat in clinical use. Moreover, from a consideration of IC50, and yield of each compound, it was concluded that tanshinone IIA (6) is the major active constituent of the MeOH extract and danshenol A (I) and (-)danshexinkun A(7) are the minor ones.

Hepatoprotective effect of Hovenia dulcis THUNB. on experimental liver injuries induced by carbon tetrachloride or D-galactosamine/lipopolysaccharide.

The hepatoprotective effects of the fruits of Hovenia dulcis THUNB. on chemically or immunologically induced experimental liver injury models were examined. The methanol extract showed significant hepatoprotective activity against CCl4-toxicity in rats and D-galactosamine (D-GalN)/lipopolysaccharide-induced liver injury in mice. The methanol extract also significantly protected against CCl4-toxicity in primary cultured rat hepatocytes. Hepatoprotective activity-guided fractionation and chemical analysis led to the isolation of an active constituent, (+)-ampelopsin (1) from the methanol extract.

Preventive effect of lithospermate B from Salvia miltiorhiza on experimental hepatitis induced by carbon tetrachloride or D-galactosamine/lipopolysaccharide.

The water extract from the root of Salvia miltiorhiza Bunge showed a protective effect on cultured rat hepatocytes against carbon tetrachloride (CCl4)-induced necrosis. A further study was carried out to isolate the active constituent. Activity guided fractionation of the extract and chemical analysis gave us lithospermate B (a salt of lithospermic acid B), a tetramer of caffeic acid. Lithospermate B was also found to have a potent hepatoprotective activity in not only in vitro but also in vivo experimental liver injuries induced by CCl4 or D-galactosamine (D-GalN)/lipopolysaccharide (LPS).