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[Quantitative analysis of deacylgymnemic acid by high-performance liquid chromatography].

A method of the quantitative analysis was established for the determination of deacylgymnemic acid (DAGA) in the alkaline hydrolysate of the sample containing gymnemic acids which are ingredients of Gymnema sylvestre R. BR. leaves, by means of high-performance liquid chromatography. This method was used for comparing the contents of gymnemic acids in various samples. The amount of gymnemic acids analyzed as DAGA in 70% ethanol extract of dry leaves was about twice that in hot water extract. The commercial health-supplemental foods of five companies were investigated for the contents of gymnemic acids as DAGA and there were large differences from 38 to 251 mg in the dosage per day recommended by each company.

Chromatography, High Pressure Liquid↗

Promotive effect of lysozyme on the ciliary activity of the human nasal mucosa.

Lysozyme is an important resistance factor in the respiratory defence mechanism. Few reports exist concerning its effects on the ciliary activity of the human nasal mucosa. To clarify the biological activity of lysozyme in ciliary movement, a quantitative study was undertaken using a photo-electrical method to measure ciliary beats in vitro under conditions of constant temperature. Egg-white lysozyme in a 0.01 M sodium phosphate buffer solution, pH 7.4, dose-dependently accelerated the ciliary beats in the human nasal mucosa, removing overlying mucus continuously for at least 60 min. The buffer alone, however, failed to affect ciliary beats. These results indicate that egg-white lysozyme directly promotes the ciliary beats in the human nasal mucosa.

Animals↗

Inhibition of RNA nucleo-cytoplasmic translocation by anti-nucleus antibody.

Anti-nucleus antibody was raised by immunizing rabbits with rat liver nuclei, and purified by affinity-column chromatography. When purified anti-nucleus IgG molecules were introduced into FL cells by the erythrocyte-ghost fusion method with HVJ (Sendai virus), release of RNA from the nucleus into the cytoplasm was inhibited in the presence of alpha-amanitin, but nuclear accumulation of 125I-labeled non-histone chromosomal protein from the cytoplasm was not inhibited. These findings suggest that the nucleo-cytoplasmic transport mechanisms of RNA and nuclear proteins are different. The molecular weight of the antigen of this antibody was determined to be about 55K by the immunoblotting technique.

Amanitins↗

Membrane topology of aspartate:alanine antiporter AspT from Comamonas testosteroni.

We cloned the aspT gene encoding the L-aspartate:L-alanine antiporter AspTCt in Comamonas testosteroni genomic DNA. Analysis of the nucleotide sequence revealed that C. testosteroni has an asp operon containing aspT upstream of the l-aspartate 4-decarboxylase gene, and that the gene order of the asp operon of C. testosteroni is the inverse of that of Tetragenococcus halophilus. We used proteoliposomes to confirm the transport processes of AspTCt. To elucidate the two-dimensional structure of AspTCt, we analysed its membrane topology by means of alkaline phosphatase (PhoA) and beta-lactamase (BlaM) fusion methods. The fusion analyses revealed that AspTCt has seven transmembrane segments (TMs), a large cytoplasmic loop containing approximately 200 amino acid residues between TM4 and TM5, a cytoplasmic N-terminus, and a periplasmic C-terminus. These results suggest that the orientation of the N-terminus of AspTCt differs from that of tetragenococcal AspT, even though these two AspT orthologues catalyse the same transport reactions.

Alanine↗

HPLC quantitative analysis of plantaginin in Shazenso (Plantago asiatica L.) extracts and isolation of plantamajoside.

Quantitative analytical method by means of high-performance liquid chromatography (HPLC) has been developed for the quality control of Shazenso (Plantago asiatica L.) extracts. Plantaginin (I) which is one of the known constituents of Shazenso, was selected as a standard compound of the analysis. A chemically labile component has been isolated from the methanol extract and identified to be plantamajoside (II). It was found that II was converted into III even in hot water.

Chromatography, High Pressure Liquid↗

[Subrenal capsule assay for testing the effectiveness of anticancer drugs].

This study compared the histological changes induced by human bladder tumor xenografts following their implantation in the subrenal capsular space of immunocompetent mice (CDF1, 6-10 weeks age). Ten human bladder tumors were studied by light microscopic and electron microscopic methods. All materials were obtained from cold-cup biopsy using endoscopic procedure. Single fragments (1 mm X 1 mm X 1 mm) were implanted on Day 0 under the renal capsule of each mouse. Animals were serially sacrificed from day 2 to day 6 and in some cases as late as day 10. All kidneys were sectioned up to 3 levels at the xenograft and stained with appropriate histological reagents. The results of this study were; 1) infiltration of the tumor with mononuclear cells and neutrophils began on day 4, 2) pretreatment with 200 mg/kg cyclophosphamide before implantation reduced cell infiltration into the xenograft, 3) complete rejection of the tumor on day 8, 4) electron microscopic study showed minimum changes of intracellular organs on day 2, 5) but microscopic architecture was preserved in the tumor explants removed on day 4. This morphological study suggested that histological evaluation on day 4 might be a suitable method of subrenal capsule assay using immunocompetent mice. We therefore tested genitourinary carcinomas to determine their sensitivities to commonly use chemotherapeutic agents using histological criteria. Groups of tumor-bearing mice were treated daily for 3 days. Sensitivities for assays were assessed on day 4 by comparison of histological score. Of the 15 cases evaluated, 20% demonstrated significant oncolytic activity. These data also suggested that a clinically relevant chemosensitivity assay might enhance the selection of appropriate therapy.

Animals↗

Length of the ribs in patients with idiopathic scoliosis.

There have been few reports of actual measurements of rib length in patients with idiopathic scoliosis. We measured the lengths of the 7th to 12th ribs in 28 patients with idiopathic scoliosis with right convex single thoracic curve. The measurements were performed by the multi-projection volume reconstruction method of computed tomography. A significant difference between the left and right side length (laterality) was observed in the 11th and 12th ribs. Laterality was not observed in the 7th, 8th, 9th, and 10th ribs. In this study, it appeared inadequate to conclude that the laterality of rib length caused the scoliosis. Since the 11th and 12th ribs were floating ribs, we speculated that, on the contrary, the scoliosis affected the laterality of the rib length.

Adolescent↗

Resource allocation in public hospitals: is it effective?

The allocative inefficiency is a fundamental flaw in the public hospitals of the developing countries. The inefficiencies drain the limited public resources allotted for healthcare. Sri Lanka's public health system faces worsening budget constraints. The resource allocation practices of the Ministry of Health focus on increasing the cadre of hospital staff, consequently crowding out the investments on facility development. The purpose of the study is to assess the impact of resource allocation in the tertiary-care public hospitals that are under the central Ministry of Health. The model is based on the assumption that the hospital managers and other agents of a public hospital pursue the objective of quality maximization (in the absence of a profit motive). The inpatient mortality rate is selected as the indicator of quality. With the use of panel data fixed-effects, and first-differencing estimation methods, we study the impact of the resource allocation on the hospital mortality rates. The selected models are statistically significant at 0.1% level. The elasticity effect of the capital is considerably larger than the effects of the human resources, in servicing the patients. The results suggest that the human resource utilization is suboptimal, due to the inadequacy of the capital (i.e. medical equipment, etc.). The reorientation of the resource allocation towards the capital investments may save more lives.

Efficiency, Organizational↗

Clinical utility of ursodeoxycholic acid in preventing flutamide-induced hepatopathy in patients with prostate cancer: a preliminary study.

BACKGROUND: The present study was designed to ascertain retrospectively the validity of ursodeoxycholic acid (UDCA) in the treatment of prostate cancer in terms of prophylactic effects on the occurrence of flutamide-induced hepatopathy in a large number of patients surveyed in a multi-center cooperative study. METHODS: One hundred and eighty-one patients (74.1 +/- 4.9 years) with prostate cancer treated with flutamide with (n = 70) or without (n = 111) UDCA were retrospectively evaluated and the occurrence of hepatopathy was compared between these two patient groups. RESULTS: Between patients treated with UDCA and those without it, no significant differences were noted in age, clinical stage, grade, duration of flutamide administration and serum prostate-specific antigen (PSA) levels before treatment. However, there were significant differences in the presence or absence of previous treatments and treatments used together with flutamide. The incidence of hepatopathy was 11.4% (8/70) in patients with UDCA and 32.4% (36/111) in those without it, showing a statistically significant difference (P < 0.05). The hepatopathy-free rate obtained by the Kaplan-Meier method was also significantly higher in patients with UDCA (88.4% 1 year following flutamide administration) than that in those without it (59.6%) (P < 0.005). CONCLUSION: These results suggest that UDCA has a prophylactic effect against flutamide-induced hepatopathy in patients with prostate cancer.

Aged↗

Input-output analysis of in vivo photoassimilate translocation using Positron-Emitting Tracer Imaging System (PETIS) data.

The Positron-Emitting Tracer Imaging System (PETIS) is introduced for monitoring the distribution of (11)C-labelled photoassimilates in Sorghum. The obtained two-dimensional image data were quantitatively analysed using a transfer function analysis approach. While one half of a Sorghum root in a split root system was treated with either 0, 100, or 500 mM NaCl dissolved in the nutrient solution, tracer images of the root halves and the lower stem section were recorded using PETIS. From the observed tracer levels, parameters were estimated, from which the mean speed of tracer transport and the proportion of tracer moved between specified image positions were deduced. Transport speed varied between 0.7 and 1.8 cm min(-1) with the difference depending on which part of the stem was involved. When data were collected in the lowest 0.5-1 cm of the stem, which included the point where the roots emerge, transport speed was less. Rapid changes in NaCl concentration, from 0 to 100 mM, resulted in short-term increases of assimilate import into the treated root. This response represented a transient osmotic effect, that was compensated for in the medium-term by osmotic adaptation. Higher concentrations of NaCl (500 mM) resulted in distinctly less photoassimilate transport into the treated root half. The present results agree with earlier observations, showing that transport of (11)C-labelled photoassimilates measured with the PETIS detector system can be quantified using the method of input-output analysis. It is worth noting that with the PETIS detector system, areas of interest do not need to be defined until after data collection. This means that unexpected behaviour of a plant organ will be seen, which is not necessarily the case with conventional detector systems looking at predefined areas of interest.

Biological Transport↗

Modified extravesical ureteroneocystostomy for completely duplicated ureters in renal transplantation.

INTRODUCTION: Completely duplicated ureters are the most common congenital malformation of the upper urinary tract. However, there are very few reports on transplantation using kidneys with double ureters, and the technique of ureterocystoneostomy for completely duplicated ureters has not yet been established. PATIENTS AND METHODS: We treated 3 patients with completely duplicated ureters at our institutions from January 1998 to October 2005. We modified the extravesical technique for treating these 3 cases and evaluated the 52-month follow-up period for possible urological complications. RESULTS: Neither urological complications nor urinary tract infections occurred in the follow-up period after using our new technique. CONCLUSION: We conclude that this technique is an appropriate procedure for the transplantation of kidneys with completely duplicated ureters.

Cystostomy↗

Polyoncogenicity and insulinoma-inducing ability of BK Virus, a human Papovavirus, in Syrian golden hamsters.

Newborn hamsters were inoculated intracerebrally with a series of purified and concentrated BK virus samples originating from a single stock of Gardner's original strain. Most (60-100%) of the hamsters developed various tumors 3-9 months later. The frequent types of tumors were ventricular tumors (choroid plexus papillomas and ependymomas: 7-53%), malignant insulinomas (0-92%), and osteosarcomas (0-50%). The T-antigen was positive in 59 of 60 tumors tested, but the virus was rescued by the cell fusion method from only 1 of 11 cell lines derived from these tumors. The incidence of insulinomas varied greatly with the virus sample; the two samples that showed the highest incidence (47 and 92%) originated from one parental virus stock, and all the other samples with the lower incidences (0-9%) originated from another ancestral stock. These results suggest the presence of a BK virus mutant(s) differing in capacity to induce insulinoma. A functional insulinoma cell line was thus established.

Adenoma, Islet Cell↗

Effect of anti-ICAM-1 and anti-LFA-1 antibodies on rat liver transplantation.

The effect of the anti-cell adhesion molecule antibodies, anti-ICAM-1 (1A29) and anti-LFA-1 (WT1), on rat liver transplantation was investigated. Livers from ACI rats were transplanted into Lewis rats by Kamada's method and during the recipient operation 1A29 1 mg/kg, and WT1 1 mg/kg were administered intravenously to one group of rats (treated group). The survival time of the treated group was significantly longer than that of the untreated group, but permanent unresponsiveness could not be induced. Postmortem examination revealed little histological evidence of acute rejection in treated rats, in which the main cause of death was thought to be chronic rejection.

Animals↗

Immunoelectron microscopic demonstration of S-100b protein-like in centriole, cilia, and basal body.

We report here the ultrastructural localization of S-100b protein-like immunoreactivity in the centriole, cilia, and basal body. Duodenum and trachea of guinea pigs and rats were fixed and immunostained by the protein A-gold method. All centrioles, cilia, and basal bodies observed showed clear S-100b protein-like immunoreactivity. Specific colloidal gold particles were located over the microtubules in these cell organelles. However, other microtubules scattered throughout the cytoplasm were devoid of immunoreactivity. Although the functional significance of S-100b protein-like immunoreactivity in the centriole, cilia, and basal bodies remains to be elucidated, the present results introduce new perspectives into the investigation of localization and function of S-100 proteins.

Animals↗

A rapid vapor-phase acid (hydrochloric acid and trifluoroacetic acid) hydrolysis of peptide and protein.

A new method for the acid hydrolysis of protein is presented. Peptide bonds are cleaved by the action of an HCl/trifluoroacetic acid (TFA) vapor mixture. Contamination for the hydrolysis mixture is reduced to low levels (1-3 pmol). Recovery of hydrophobic amino acid is improved. Short reaction times are achieved and rapid removal of acids is facilitated. The reaction temperature is 158 degrees C for reaction times of 22.5 and 45 min with 7 M HCl and 10% TFA containing 0.1% phenol.

Amino Acids↗

Evaluation of sperm fertilizing capacity using the determination of acrosome reaction.

To evaluate the usefulness of the determination of acrosome reaction (AR) at 2 and 4 hours of incubation for predicting sperm fertilizing capacity in vitro, semen samples obtained from 45 partners of couples participating in the IVF (in vitro fertilization) program were examined. The population was divided into 2 groups, depending on the fertilization rate in the IVF program: Group A, with a good fertilizing capacity (> or = 50% fertilized oocytes); and Group B, with a poor fertilizing capacity (< 50% fertilized oocytes). The rate of acrosome reaction (%AR) was examined using the fluorescein isothiocyanate-conjugated Concanavalin A lectin method. The efficacy of this procedure was compared with that of the zona-free hamster oocyte human sperm penetration assay (SPA) and sperm characteristics (SC) that appear in routine semen analysis. The sensitivities in the test of AR, SPA and SC were 91%, 86% and 82%, respectively. However, their specificities were 83%, 48% and 43%, respectively. Therefore, the screening procedure using the determination of acrosome reaction might be a useful tool in predicting IVF results.

Acrosome↗

A comparison of period amplitude analysis and FFT power spectral analysis of all-night human sleep EEG.

Zero-cross and zero-derivative period amplitude analysis (PAA) data were compared with power spectral analysis (PSA) data obtained with the fast Fourier transform in all-night sleep EEG from 10 subjects. Although PAA zero-cross-integrated amplitude showed good agreement with PSA power in 0.3-2 Hz, zero-cross analysis appears relatively ineffective in measuring 2-4 Hz and above waves. However, PAA zero-derivative measures of peak-trough amplitude correlated well with PSA power in 2-4 Hz. Thus, while PAA appears able to measure the entire EEG spectrum, the analytic technique should be changed from zero cross to zero derivative at about 2 Hz in human sleep EEG. PAA and PSA both demonstrate robust and interrelated across-night oscillations in three frequency bands: delta (0.3-4 Hz); sigma (12-16 Hz); and fast beta (20-10 Hz). The frequencies between delta and sigma, and between sigma and fast beta, did not show clear across-night oscillations using either method, and the two methods showed lower epoch-to-epoch agreement in these intermediate bands. The causes of this reduced agreement are not immediately clear, nor is it obvious which method gives more valid results. We believe that the three strongly oscillating frequency bands represent fundamental properties of the human sleep EEG that provide important clues to underlying physiological mechanisms. These mechanisms are more likely to be understood if their dynamic properties are preserved and measured naturalistically rather than being forced into arbitrary sleep stages or procrustean models. Both PAA and PSA can be employed for such naturalistic studies. PSA has the advantages of applying the same analytic method across the EEG spectrum and rests on more fully developed theory. Combined zero-cross and zero-derivative PAA demonstrates EEG oscillations that closely parallel those observed with spectral power, and the PAA measures do not rely on assumptions about the spectral composition of the signal. In addition, both PAA techniques can measure the relative contributions of wave amplitude and incidence to total power: These waveform characteristics represent different biological processes and respond differentially to a wide range of experimental conditions.

Adult↗

[Development for the determination of band 5b tartrate-resistant acid phosphatase activity in serum].

Band 5 Tartrate-resistant acid phosphatase(TRACP; EC 3.1.3.2) consists of two isoenzymes, bands 5a and 5b, of which band 5b TRACP, an enzyme expressed in bone-resorbing osteoclasts, is secreted into the circulation during bone resorption. Band 5b TRACP was measured kinetically in serum as tartrate-resistant fluoride-sensitive heparin-resistant ACP with 2,6-dichloro-4-acetylphenyl phosphate as substrate at pH6.6. The within-run(n = 20) and between-run(n = 20) CVs of band 5b TRACP activity were 3.3-5.8% and 5.0-7.3%, respectively. The reference range of band 5b TRACP activity in males(n = 72) and females(n = 87) 20-39 years of age by this method were 3.7-12.5 U/l and 2.7-9.9 U/l, respectively. The band 5b TRACP value was significantly higher in post-menopausal women compared with the menstruating women. The relationship of band 5b TRACP and ultrasound findings in healthy women aged 31-75 years(n = 139) were inversely correlated with stiffness(r = -0.401), speed of sound(SOS; r = -0.386) and broadband ultrasound attenuation(BUA; r = -0.338). These results suggest that band 5b TRACP may be a useful in the evaluation of bone turnover.

Acid Phosphatase↗