An Immunoenzyme Procedure for Enumerating Fibronectin-Secreting Cells

A recently described solid phase immunoenzyme procedure (ELISPOT) has been adapted for the detection of individual cells secreting fibronectin. Simple and sensitive, this technique should find useful application for studying fibronectin production at the cellular level.     

Female rats do not exhibit free fatty acid-induced insulin resistance

It is well described that excessive lipid metabolism can cause insulin resistance in both animals and humans, and this has been implicated as a causative factor in the development of insulin resistance and type 2 diabetes in humans. Recently, we have shown that intravenous lipid emulsion (liposyn) infusion during a 120-min euglycemic-hyperinsulinemic clamp led to significant reductions in insulin action and fatty acid translocase (FAT/CD36) skeletal muscle protein expression. After reviewing the literature, it became evident that essentially all past studies, including our own, were conducted in male animals. Therefore, to determine whether there were sex determinants of fat-induced insulin resistance, we assessed the impact of free fatty acid (FFA) elevation on insulin action in female rats. Here, we report that a fourfold elevation in plasma FFA concentration induced a 40% reduction in the insulin-stimulated glucose disposal rate, a 30% decline in insulin-stimulated skeletal muscle insulin substrate receptor-1 (IRS-1) phosphorylation, a 48% decrease in IRS-1-associated phosphatidylinositol (PI) 3-kinase activity, and a 50% reduction in muscle FAT/CD36 protein expression in male rats. In striking contrast, we found no effect of FFA elevation to cause insulin resistance, changes in IRS-1/PI 3-kinase, or FAT/CD36 protein levels in female animals. Our findings indicate that female animals are protected from lipid-induced reductions in insulin action.     

First human trial of FTY720, a novel immunomodulator, in stable renal transplant patients

FTY720 is a novel immunomodulator to be developed for use in organ transplantation. The primary objective of this study was to measure safety, single-dose pharmacokinetics, and pharmacodynamics in stable renal transplant patients-the first human use of FTY720. This study used a randomized, double-blind, placebo-controlled design that explored single oral doses of FTY720 from 0.25 to 3.5 mg in 20 stable renal transplant patients on a cyclosporine-based regimen. Safety assessments and blood samples were taken predose and at multiple time points during a 96-h period postdose. Standard pharmacokinetic parameters were derived from the FTY720 whole blood concentrations, measured by HPLC/MS/MS. FTY720 was well tolerated, with no serious adverse events. Transient, asymptomatic bradycardia occurred after administration in 10 of 24 doses of FTY720. Pharmacokinetics are characterized by a prolonged absorption phase; the terminal elimination phase started 36 h after the administration, with elimination half-life (t(1/2)) ranging from 89 to 157 h independent of dose. Maximum plasma concentration and AUC were proportional to dose with low intersubject variability, the apparent volume of distribution (V(d)/F) ranged from 1116 to 1737 L. FTY pharmacodynamics were characterized by a reversible transient lymphopenia within 6 h, the nadir being 42% of baseline. The lymphocyte count returned to baseline within 72 h in all dosing cohorts except the highest. Single oral doses of FTY720 ranging from 0.25 to 3.5 mg were well tolerated and caused a reversible selective lymphopenia. Transient, but asymptomatic bradycardia was the most common adverse event. The long t(1/2) suggests less frequent dosing intervals. The size of V(d)/F is in excess of blood volume, consistent with widespread tissue distribution     

A dropout analysis of the second phase of the Swedish part of the European Youth Heart Study

The European Youth Heart Study (EYHS) addresses cardiovascular disease risk factors and their determinants in European children and adolescents. The Swedish part of the study began with cross-sectional data collection in 9- and 15-year-old schoolchildren in 1998–1999 (EYHS-I). Repeated observations of the key indicators were performed in 2004–2005 (EYHS-II). The purpose of this study was to assess potential dropout effects in EYHS-II. Participants in both EYHS-I and EYHS-II (n=459) were compared with dropouts who participated only in the EYHS-I (n=678) in relation to baseline physical activity, cardiorespiratory fitness and socioeconomic and anthropometric characteristics. Bivariate comparisons were performed using chi-square tests and gamma tests for nominal and ordinal data, respectively. Continuous data were compared by t tests and Mann−Whitney tests depending on the distribution. The Bonferroni correction was used to control for multiple hypothesis testing. Multiple logistic regression with backward elimination of variables was applied to study independent effects of variables on the probability of becoming a dropout. Analyses were performed separately for the younger and older age groups. The dropout proportion in EYHS-II was 60%. Subjects from the older age group were less likely to participate in the follow-up study (32% vs. 50%, p<0.001). In bivariate analyses, only maternal education was associated with dropout rates in the younger age group after Bonferroni correction. Males were more likely to drop out in both younger [odds ratio (OR)=1.72; 95% confidence interval (CI): 1.10, 2.96] and older (OR=1.96; 95% CI: 1.09, 3.54) age groups while basic maternal education was associated with outcome only in the younger group (OR=4.31; 95% CI: 1.78, 2.95) in regression analysis. The Swedish EYHS-II had high dropout rate after EYHS-I, but the dropouts did not differ from the participants in relation to physical activity, physical fitness, and anthropometric indices. Males were more likely to drop out than were females in both age groups. Differential dropout in relation to maternal education was observed in the younger age group.     

Cancer incidence in the native peoples of far eastern siberia

Cancer incidence rates were examined in the native peoples of the far north-east of Siberia for the years 1977–1988. Particularly high rates of cancers of the stomach, lung, oesophagus and cervix were observed. For stomach cancer, the male and female age-standardized (to the world population) rates were 103.9 per 100,000 and 50.0 per 100,000 respectively. The corresponding lung cancer rates were 109.4 and 45.7, and for oesophageal cancer 83.9 and 35.0. The age-standardized cervical cancer rate was 38.5 per 100,000. Rates of these cancers were considerably higher than in native Alaskan peoples, although the latter had higher rates of breast and colorectal cancers. The rates were also much higher than those of the migrant peoples from Russia and elsewhere who have settled in the far north-east over the past 3 centuries, particularly at younger ages. Male rates of stomach and lung cancer were highest in the paleo-Asiatic peoples of the north, whereas male oesophageal rates were highest in the Taiga people. In females, rates of stomach and oesophageal cancers were highest in the paleo-Asiatic peoples, and rates of lung cancer were highest in the Taiga nationalities. Cervical cancer rates were highest in the Amuro-Sakhalin nationalities of the south. Further research is needed at individual levels to explain the very high risks and the differences among the ethnic groups.     

A predictive role of neoangiogenesis and comparison with other prognostic factors

Incidental prostatic carcinoma (ICP) has good prognosis related to low stage at diagnosis. Few rogressive cases demanding aggressive treatment need early identification. Neoangiogenesis proved its predictive role in prostatic carcinoma after radical prostatectomy. To reveal its value in ICP authors investigated specimens after transurethral resection of prostate (TURP). Retrospective study was performed on 68 ICP diagnosed in years 1985 1989. Microvessels highlighted by factor VIII were counted in a x200 microscope field (0,8012 mm 2 ) in most active areas of neovascularisation. Microvessel count was correlated with tumor differentiation degree, Gleason score, disease stage, and patients survival in at least 9 years after diagnosis. Higher maximal microvessel counts were associated with lower degree of tumor differentiation (p=0,005), Gleason score (p=0,001), and disease stage (0,003). No association with disease progression and patients survival was found. Mean microvessel counts showed less significant values when correlated with tumor differentiation degree (p=0,003) and Gleason score (p=0, 01), and no correlation with other variables. Microvessel density in TURP specimens of ICP retains its prognostic value already demonstrated in carcinoma of peripheral prostatic lobes. Maximal microvessel counts were prognostically more reliable than mean values.     

Studies on the mechanism of dog heart (Na+ + K+)-ATPase inhibition by D,L-aspartic acid and its K+ and Mg2+ salts.

d,l-aspartic acid as well as its K⁺ and Mg²⁺ salts have been found to be inhibitory to Na⁺ + K⁺)-ATPase activity of the dog heart microsomal fraction enriched with sarcolemma. According to intensity of the inhibitory effect (noncompetitive) they form the following sequel: mono-K⁺-aspartate $\text{?}\text{d,l}\text{-aspartic acid}\text{<}\text{mono-Mg}{}^{\mathrm{2+}}\text{-}\text{aspartate}\text{<}\text{mixture}$ of K⁺ and Mg²⁺-aspartate in a molar ratio of 1:1 (K⁺, Mg²⁺-ASP). The latter mixture is widely used as an agent in cardiac failure. For (Na⁺ + K⁺)-ATPase the salient effects of d,l-aspartic acid and/or its K⁺ and Mg²⁺ salts were: (i) decrease in V for ATP as substrate with unchanged K_m; (ii) for Na⁺ as an allosteric modifier of (Na⁺ + K⁺)-ATPase activity a decrease in V without any alteration in n as a measure of cooperativity between activating sites; (iii) for K⁺ a decrease in V and n as well as an increase in K_0.5. In the presence of Na⁺ and ATP the high affinity of the enzyme for K⁺ became reduced by d,l-aspartic acid, lowering at the same time the K_0.5 value.Effects like these have also been described for ouabain. The present data show that K⁺ and Mg²⁺ salts of d,l-aspartic acid act at a similar locus as does ouabain.     

Cervids as Babesiae hosts, Slovenia

We describe cervids as potential reservoir hosts of Babesia EU1 and B. divergens. Both babesial parasites were found in roe deer. Sequence analysis of 18S rRNA showed 99.7% identity of roe deer Babesia EU1 with the human EU1 strain. B. divergens detected in cervids was 99.6% identical to bovine B. divergens.     

Determination of some organochlorine compounds in herbal colouring agent henna (Lawsonia inermis) and in tea (Thea sinensis)

Henna (Lawsonia inermis) has been used for centuries as a herbal hair and skin dye, but very little is known about its additives and contaminants that could adversely affect human health. An analytical method was developed to determine organochlorine compounds in henna, as they are still widely used in the areas where henna is grown. Samples were sonicated with n-hexane, extracts cleansed on Florisil sorbent and analysed using gas chromatography with electron capture detection. The overall recoveries were 17-33 % with the extraction RSD 5-21%, while the levels of lindane (gamma-HCH), p,p'-DDT, and p,p'-DDE in henna samples were 7-157 microg kg(-1). The same procedure was successfully applied to analyse black tea samples for the same compounds, and which showed lower contamination.     

Dissociation of the metal inlay from the polyethylene liner in an uncemented threaded cup

We describe a case of spontaneous dissociation of the metal inlay from the polyethylene cup of a sandwiched metal-on-metal total hip prosthesis manufactured from Co-28Cr-6Mo SM 21 alloy. The patient, a 51-year-old active woman, started to feel groin pain 50 months postoperatively. The pain progressed after a slight trauma and led to final revision after 66 months in situ. No signs of impingement were observed. The polyethylene cup showed an approximately 2 mm-deep groove in the superior wall, and the head was heavily worn on the lateral side. In addition to the typically observed abrasive wear patterns, several types of severe wear defects were noticed. Extensive metallosis and necrosis were observed histologically. This unusual case of substantial deformation of the head and the cup was presumed to have occurred as a result of the increased friction and consequent high wear of the metal head.