Quantitative Analysis of Dammarane-type Ginsenosides in Different Ginseng Products

Ginseng products available in different forms and preparations are reported to have varied bioactivities and chemical compositions. In our previous study, four new dammarane-type ginsenosides were isolated from Panax ginseng, which are ginsenoside Rg18 (1), 6-acetyl ginsenoside Rg3 (2), ginsenoside Rs11 (3), and ginsenoside Re7 (4). Accordingly, the goal of this study was to determine the distribution and content of these newly characterized ginsenosides in different ginseng products. The content of compounds 1 – 4 in different ginseng products was determined via HPLC-UV. The samples included ginseng roots from different ginseng species, roots harvested from different localities in Korea, and samples harvested at different cultivation ages and processed under different manufacturing methods. The four ginsenosides were present at varying concentrations in the different ginseng samples examined. The variations in their content could be attributed to species variation, and differences in cultivation conditions and manufacturing methods. The total concentration of compounds 1 – 4 were highest in ginseng obtained from Geumsan (185 µg/g), white-6 yr ginseng (150 µg/g), and P. quinquefolius (186 µg/g). The results of this study provide a basis for the optimization of cultivation conditions and manufacturing methods to maximize the yield of the four new ginsenosides in ginseng.

Acer okamotoanum Inhibit the Hydrogen Peroxide-Induced Oxidative Stress in C6 Glial Cells

Chronic oxidative stress due to the accumulation of reactive oxygen species (ROS) in neuronal cells ultimately leads to neurodegenerative diseases. The use of natural therapies for the prevention of ROS-induced cell damage and for the treatment of neurodegenerative disorders has shown promising results. In this study, we evaluated the neuroprotective effects of the ethyl acetate (EtOAc) fraction of A. Okamotoanum against the hydrogen peroxide (H₂O₂)-induced oxidative stress in C6 glial cells. Results show that cell viability was decreased in cells incubated with H₂O₂, whereas the addition of EtOAc fraction treatments in such cells significantly increased viability. The EtOAc fraction showed the highest inhibitory activity against ROS production and it also decreased the expressions of inflammatory proteins including cyclooxygenase-2, inducible nitric oxide synthase and interleukin-1β. Furthermore, the EtOAc fraction inhibited apoptosis by regulating the protein expressions cleaved caspase

Optimization of Extraction Conditions and Quantitative Analysis of Isoquercitrin and Caffeic Acid from Aster scaber

To determine the optimum extraction conditions that give the highest yield of isoquercitrin and caffeic acid from Aster scaber, the effects of four extraction variables (solvent concentrations, extraction time, number of repeated extraction, and solvent volumes) on isoquercitrin and caffeic acid yield was examined via HPLC-UV. Our results showed that the highest extract and isoquercitrin yield were observed when A. scaber was extracted with 450 mL distilled water for 8 hr repeatedly for three times. In case of caffeic acid, the content was higher in the two repeated extracts. Also, content analysis of isoquercitrin in Aster species was performed in which A. fastigiatus, A. ageratoides, and A. scaber exhibited the highest isoquercitrin content at 6.39, 5.68, and 2.79 mg/g extract, respectively. In case of caffeic acid, the highest content of A. scaber and A. glehni was 0.64 and 0.56 mg/g extract, respectively. This study reports an optimized method for extraction of isoquercitrin and caffeic acid from A. scaber and evaluates potential sources of the compounds.

Determination of Silybin B in the Different Parts of Silybum marianum using HPLC-UV

Silymarin is the standardized extract from Silybum marianum which consists mainly of flavonoids and polyphenols. It is highly regarded for its hepatoprotective ability. Silybin B is a flavonolignan and one of the active components of silymarin. The content of silybin B in various parts of S. marianum was analyzed by HPLC-UV. Results show that the extract of seeds contain the highest amount of silybin B (7.434 mg/g DW). The petioles of S. marianum showed a low content of silybin B. This study revealed that seeds of S. marianum contain high amount of silybin B and could be a good source of the compound.

Content Analysis of Rutin in the Leaves of Boehmeria nivea Harvested in Different Regions of South Korea by HPLC-UV

Phytochemical analysis of Boehmeria nivea (Bn) leaves by medium pressure liquid chromatography led to the isolation of a flavonoid glycoside identified by spectroscopic analysis as rutin. The amount of rutin in the leaves of Bn harvested from nine regions in South Korea (Bn 1–9) which were collected on the months of June, July, August, and September was determined by HPLC-UV analysis. A gradient elution program that utilizes a Discovery® C18 (4.6 × 250 mm, 5 µm) column and mobile phase composed of 1% acetic acid-water: acetonitrile (90:10 to 60:40 for min) was followed. The injection volume and flow rate were 10 µl and 1 mL/ min, respectively. UV detection was set at 350 nm. Results show that Bn-8 harvested in September reported the highest content of rutin among the samples analyzed. This study provides a basis for the optimal harvest time of Bn which maximizes the yield of rutin.

Quantitative Determination of Bakkenolide D in Petasites japonicus and Farfugium japonicum by HPLC/UV

A quantitative analysis of bakkenolide D in the different parts of Petasites japonicus and Farfugium japonicum was performed by HPLC. A gradient HPLC elution system with a mobile phase consisting of water:acetonitrile solution (20:80 to 0:100 for 45 min) was followed and an INNO C₁₈ column was used for the chromatographic separation. The injection volume, flow rate, and UV detection were 10 µL, 1 mL/min, and 290 nm, respectively. Results show that both species showed the highest amount of bakkenolide D in the roots being 107.203 and 166.103 mg/g for P. japonicas and F. japonicum, respectively. Content analysis on the different parts of both plants displayed remarkably lower values which ranged from 0.403 – 4.419 and 7.252 – 32.614 mg/g for P. japonicas and F. japonicum, respectively. The results show that the roots of both plants are rich in bakkenolide D showing a promising use in the development of nutraceuticals and industrial application of the compound.

Trends in leprosy over fifty years in Gudiyatham Taluk, Vellore, Tamil Nadu.

This paper presents epidemiological trends over a fifty-year period observed in a defined population served by the Schieffelin Leprosy Research and Training Centre (SLR & TC), Karigiri, Vellore District in Tamil Nadu. It covers three distinct periods, namely, the pre-MDT era with dapsone monotherapy, the MDT era under a vertical leprosy control programme and the MDT era after leprosy control services were integrated into the general health services. Prevalence rates have declined steadily from 125 per 10,000 population at the time of introduction of MDT in 1982 to 5 per 10,000 at the time of integration in 1997 to less than 1 per 10,000 in 2005. The new case-detection rate was 5.4 per 10,000 when the field programme started in 1962, and held steady at 15-20 per 10,000 between 1970 and 1980. It then showed a gradual fall from 10.8 per 10,000 in 1985 to 3.9 at the time of integration, and continued to fall in the post-integration period and was 0.8 per 10,000 in 2005. The mean age at detection showed a gradual increase from 23.4 years in the dapsone era to 31.2 years in the post-integration period. The male: female ratio showed a preponderance of males almost throughout the reference period. While polar types of leprosy (TT & LL) were common in the dapsone era, more of borderline leprosy (BT & BL) cases was seen more recently. MB rates that were high initially, declined steadily during monotherapy and stabilized between 10% and 12% during the vertical MDT programme and is showing an increase in the post-integration-period. The proportion of cases with Grade 2 disability at registration showed a gradual decline during the monotherapy period, remained relatively unchanged at 8%-10% during the the MDT period, and showed a sharp rise in the immediate post-integration period before falling. Analysis of trends of leprosy in a well-defined geographical population over a fifty-year period gives useful information on how the disease has evolved over the years. It provides opportunities to explore the reasons for the changes observed, though one has to be cautious while interpreting such data due to changes in definition, the play of operational factors, and changes in policies and strategies.

Are viable Mycobacterium leprae present in lepromatous patients after completion of 12 months' and 24 months' multi-drug therapy?

A study was carried out to determine whether or not viable bacilli persist in MB patients treated with 12-month and 24-month multidrug therapy (MDT). In the first group, 60 untreated lepromatous patients who had an initial average bacterial index (BI) of 3+ or more were enrolled. At the completion of 12 months of MDT, skin biopsies were obtained and M. leprae concentrate was inoculated into the footpads of five thymectomized and irradiated (T900r) mice. Rees technique was used for the mouse footpad (MFP) experiment. Harvesting was done it the 6th, 9th and 12th months. Out of the 60 biopsies inoculated into mouse footpads to check the viability of bacilli, 2 skin biopsies (3.3%) showed significant growth and 10 (16%) showed equivocal growth. 27 patients also had nerve biopsies tested for growth in MFP studies. None of the inoculated nerve biopsies showed significant multiplication in the MFP experiments. However, 4 biopsies (14%) showed equivocal growth. In the second group, 20 patients had skin biopsies and 10 had nerve biopsies done at the end of 24 doses of MDT in order to test the viability of bacilli; none of the skin or nerve biopsies from these patients showed any growth. This study showed that M. leprae present in the tissues after 24 doses of MDT are not viable and the drug schedule of 24 doses is adequate to treat leprosy patients, irrespective of their BI. However, a small (3.3%) percentage of the patients with a high BI harbour viable bacteria in the skin after 12 doses of treatment. Since a large majority of the patients (38 patients) who had a high initial BI responded well to the treatment, it is important to find out the reason for the lack of response in two patients. One of the reasons may be the presence of drug-resistant strains. It is important to follow up on these patients for a longer duration to ascertain whether or not they would relapse.

Drug resistant-Mycobacterium leprae--results of mouse footpad studies from a laboratory in south India.

Out of 265 biopsies of leprosy patients received at the Experimental Pathology Laboratory of Schieffelin Leprosy Research and Training Centre from 1987 to 1997 for evaluating resistant strains of M. leprae, using the mouse footpad technique, 49 showed resistant strains of M leprae to varying concentrations of dapsone, rifampicin and clofazimine. 23 (47%) of these were from a control area. With 369 skin-smear positive multibacillary (MB) patients as the risk group (denominator), 23 (6.23%) were resistant to one or more drugs. 18 (4.88%) had dapsone resistance, 5 (1.36%) were resistant to rifampicin and 9 (2.44%) had resistance to low concentrations of clofazimine (0.0001%). Out of the 23 biopsies with drug resistance from the control area, primary dapsone resistance was seen in 7 (30%) biopsies and secondary dapsone resistance in 11 (48%). Primary rifampicin resistance was seen in 4 (17.4%) patients, secondary rifampicin resistance in 1 (4.35%) and primary clofazimine resistance in 7 (30%). 3 (13%) of the strains showed secondary clofazimine resistance. One biopsy had resistant strains to all the three drugs. In a control area where properly supervised effective multidrug therapy (MDT) was regularly administered over the years, the emergence of drug resistance is negligible. It may not be the case if the content, duration and regularity of the drug regimen were not satisfactory. Aware of the possible shortcomings in mass administration of MDT, it is emphasized that mouse footpad studies on drug resistance should be made available at least in endemic areas where the incidence of the disease has not changed despite good MDT coverage in order to monitor the emergence of drug resistance. Research into molecular biological identification of drug resistant-M.leprae should be intensified. These steps would help to institute timely measures to check the spread of any drug-resistant organisms in the community.

Manejo de los traumatismos de pelvis / Management of pelvic traumatisms

Antecedentes: Los traumatismos de pelvis constituyen un capítulo muy importante dentro del estudio de los pacientes politraumatizados, en ellos se pueden producir lesiones de distintas estructuras que requieren de la participación de un equipo multidisciplinario para su manejo. Objetivo: Analizar una serie de casos de traumatismos de pelvis asistidos en los últimos 10 años y actualizar los conceptos fundamentales para su mejor manejo clínico y quirúrgico. Lugar de aplicación: Servicios de Emergencias y de Cirugía General hospitalarios. Diseño: Estudio observacional retrospectivo basado en criterios básicos y descripción de hallazgos clínicos (serie de casos). Población: Muestra consecutiva de 4.212 casos de pacientes asistidos por presentar traumatismo abdómino-pélvico o de pelvis puro en el período comprendido entre el 1/1/91 y el 31/12/00. Métodos: Se analizaron las historias clínicas de los pacientes de la población en estudio para determinar el número de casos que presentaron lesiones pélvicas, y en ellos los mecanismos de producción de los traumatismos, el tipo de las lesiones observadas, las distintas conductas terapéuticas realizadas y la morbi-mortalidad observada. Resultados: Presentaron lesiones 321 casos (7,62 por ciento). Se debieron a traumatismos cerrados 249 casos (77,57 por ciento), a penetrantes 46 (14,32 por ciento) y a empalamientos 26 (8,09 por ciento). Presentaron lesiones digestivas 61 pacientes (19 por ciento), vasculares 12 (3,73 por ciento), urinarias 44 (13,70 por ciento), uterovaginales 11 (3,42 por ciento), compromiso perineal 29 (9,03 por ciento) y fracturas del anillo óseo 229 (71,33 por ciento). Se efectuó tratamiento quirúrgico en 113 casos (35,20 por ciento). La morbilidad observada fue del 20,56 por ciento y la mortalidad del 7,16 por ciento. Conclusiones: Para realizar un correcto manejo de los traumatismos de pelvis se debe seguir una adecuada sistemática de estudio y tratamiento que requiere de la participación coordinada de un equipo multidisciplinario, en el cual el Cirujano General debe cumplir un rol fundamental; siendo ésta la única manera de evitar o disminuir la alta morbi-mortalidad que pueden presentar éste tipo de traumatismos

Animal-passage of campylobacter strains with iron-dextran enhances the expression of an outer membrane protein

The outer membrane proteins [OMP] of four C. jejuni/coli isolates have been identified by SDS - PAGE using Sarkosyl extraction of crude membranes. Sarkosyl insoluble fractions of four strains consistently contained a major OMP in the range of 43 k to 48 k, in addition to some minor proteins. SDS - PAGE profile of animal - passed virulence enhanced strains, when compared with the original isogenic strains when compared with the original isogenic strains, revealed the presence of a protein band migrating at 66 k to 68 k in addition to the characteristic major OMP band, suggesting a selective enhancement of this protein during animal passage in presence of iron - dextran. This shift in OMP expression was associated with enhanced virulence of C. jejuni for BALB/c mice. Immunobloting to analyse the antigens in OMP showed antigenicity to the major OMP. The role of the 68 k OMP in the pathogenesis of C jejuni for BALB/c mice remains to be studied

An insular outbreak of dengue fever in a rural south Indian village.

In mid 1990, an epidemic of fever affected a single village in Kaniyambadi Block, South India. The illness was characterized by a fever of approximately five days duration, accompanied by headache, chills, sweating and muscle pain. The overall attack rate was 22.5 per cent. The attack rate was uniform across the various age groups and between the sexes. Testing of the acute and convalescent serum samples obtained from cases showed a serological response to dengue virus. The Aedes house index in the village was found to be 36 per cent with toilets serving as active breeding sites. Adjacent villages studied showed similarly high rates of Aedes prevalence, although no cases of the same fever were seen. Since previous exposure to dengue increases the risk for epidemics of dengue hemorrhagic fever and dengue shock syndrome, health education on methods of Aedes control is continuing.