Variable alteration of regional tissue oxygen pressure in rat hippocampus by acute swimming exercise.

UNLABELLED: One of the events in the brain is an increasing cerebral blood flow during exercise. The tissue oxygen level may be increased because blood flow correlates with tissue oxygen level. However, it is little known whether the tissue oxygen pressure in hippocampal region (Hip-pO2) will be affected by exercise. AIMS: The aim of this study is to examine Hip-pO2 levels in the hippocampus and its changes during exercise. MAIN METHODS: We applied improved Clark-type electrodes to measure Hip-pO2 level in the hippocampus of rats that were subjected to three groups, 2h swimming without weights (low intensity, n=6), 2h swimming with a 5 g weight (moderate intensity, n=6), and 2h swimming with a 10 g weight (high intensity, n=6). KEY FINDINGS: Exercise affected the Hip-pO2 level, the responses varied with the exercise intensity and duration. Interestingly during and after the Low intensity swimming the Hip-pO2 level showed long lasting enhancement (10-20% above resting level). But the moderate and high intensity swimming increased Hip-pO2 level at the start of the swimming (50%, P<0.05 and slightly above resting level, respectively, at 10 min of 2h swimming) and then began to decrease (at 120 min and 10 min of 2h swimming, respectively), and suppressed the Hip-pO2 levels during post exercise resting period (2h) (85-95% of resting level, NS and 60-70% of resting level P<0.05, respectively). SIGNIFICANCE: We propose that exercise-induced hippocampal hyper/hypo oxygen condition may participate in beneficial exercise effects on brain function.

Fetal aortic isthmus growth and morphology in late gestation.

OBJECTIVE: To establish normal values for fetal aortic isthmus diameter in late gestation and to identify any changes in aortic isthmus dimensions and morphology in pathological conditions. METHODS: In this prospective study, the fetal aortic isthmus was evaluated in 110 low-risk pregnant women at between 30 and 40 weeks of gestation and 42 pregnant women who were at high risk for congenital heart defects. From coronal echocardiographic images of the connection between the aorta and ductus arteriosus, the internal diameter of the aorta was measured at the middle of the isthmus, at the point of the isthmus just proximal to the entry of the ductus arteriosus and at the descending aorta below the entry of the ductus arteriosus. RESULTS: Correlation coefficients for the diameter of each aortic segment when related to gestational age varied from r = 0.60 to r = 0.80 (P < 0.001 for each), and growth curves were derived from the third and 97th percentiles about each linear regression analysis. The mean and the third percentile for the ratio of the isthmus just proximal to the entry of the ductus arteriosus to the middle of the isthmus were 1 and 0.81. In one fetus of the high-risk patients, a contraductal shelf and the accompanying area of tubular isthmic hypoplasia were suspected and a diagnosis of coarctation of the aorta was subsequently confirmed after birth. In two fetuses with growth restriction and one fetus with intestinal atresia, the isthmus diameters were below the third percentile but the ratios of the isthmus end to the middle of the aortic isthmus were all normal and no cardiac anomalies were detected after birth. CONCLUSIONS: We could establish normal values for aortic isthmus diameters in late gestation from a coronal view and identify even minimal changes in aortic isthmus dimensions and morphology in pathological conditions.

Stable inversion method for a polarized-lidar: analysis and simulation.

A new inversion inhomogeneous atmosphere (IA) method that is more stable than Fernald's method for two-component (molecule and aerosol) scattering analysis of polarized Mie lidar signals is proposed and examined. The backscattering coefficient and the extinction-to-backscattering ratio (EBR) can be calculated for specified regions at which the depolarization ratio is less than that of molecule without further assumptions. The inversion procedure can be extended to both inward stepwise and outward stepwise integration algorithms. Simulation results indicate that a higher precision was achieved with the IA method than with Fernald's method in terms of error and random noise in estimating boundary value and EBR. Experimental results were also better with the IA method than with Fernald's method.

A case study on Salmonella enteritidis (SE) origin at three egg-laying farms and its control with an S. enteritidis bacterin.

In the early 1990s, three egg-laying farms (farms S, T, and B) were thought to have the possibility of Salmonella enteritidis (SE) contamination because positive liquid egg samples originated from those farms. The present study was therefore conducted. The first clarification for SE contamination was the study on the origin of SE contamination including its vertical transmission. The results of SE contamination profiling with dust and manure, food materials, dead embryos, and residual yolks on hatch day in regular monitoring over a few years were clearly negative. Therefore, we concluded the SE transmission/infection was attributed to horizontal infection in the egg-laying farms but not vertical transmission from parental stock, hatcheries, growth, or food materials during a 7-yr experimental period. Second, we attempted to clarify if administration of an SE bacterin (Layermune SE) to growth flocks for the egg-laying farms could reduce SE incidence in liquid egg samples from each egg-laying farm. In the first experiment, we compared SE incidence in liquid egg samples from vaccinated and nonvaccinated flocks (similar age flocks). SE incidence from vaccinated and nonvaccinated flocks showed negative and <2 most probable number (MPN)/100 ml for farm B, <2 and >1600 MPN/100 ml for farm S, and negative and >1600 MPN/100 ml for farm T, respectively. In the second experiment, we compared the SE isolation incidence in the liquid egg samples from nonvaccinated and newly replaced vaccinated flocks in the same chicken houses from each of the three egg-laying farms. SE incidence in the liquid egg samples was similar to that in the first experiment. Therefore, the SE bacterin may play an important role in reducing the SE incidence of liquid egg samples.

Establishment of an enzyme-linked immunosorbent assay with a coated deflagellated Salmonella enteritidis antigen for detection of a specific chicken antibody.

We attempted to establish an enzyme-linked immunosorbent assay (ELISA) for field monitoring/profiling purposes for Salmonella enteritidis (SE) infection of poultry flocks. Serotyping rabbit sera, commercially obtained, specific for Salmonella identification sera to O2, O4, O7, O8, S. Vi, S. Hm, and O9, showed negative ELISA (E)-values (< 0.2) on ELISA, except the O9 identification serum (E-value > 0.5). Similar negative E-value results were obtained for antisera to Echerichia coli (E. O antigen). Field serum samples originating from SE-isolated flocks yielded similar positive ratios on both ELISAs including the present coated deflagellated SE antigen and a commercially obtained flagellated SE antigen and that of rapid plate aggregation with a pullorum antigen (PD-RPA). About 100 days after the first monitoring, no SE isolation in the same flock was observed resulting in a carrier state of SE infection. Although both the monitoring results with commercially obtained ELISA and PD-RPA showed lower positive or negative ratios, the present ELISA showed a higher positive ratio than that of the first monitoring. The present ELISA is suggested to be a suitable method to do accurate profiling on the carrier state of infection.

Low density lipoprotein (LDL) binding affinity for the LDL receptor in hyperlipoproteinemia.

We measured the binding affinity of low density lipoprotein (LDL) for the LDL receptor in patients with various types of hyperlipoproteinemia and investigated the effects of LDL lipid composition and particle size on receptor affinity. LDL (1.019 < d < 1.063) was isolated by sequential ultracentrifugation from the serum of normolipidemic controls and patients with hyperlipoproteinemia. Patients with type IIa hyperlipoproteinemia had LDL with a similar receptor affinity to that of normal LDL. However, patients with hypertriglyceridemia (type IIb and type IV hyperlipoproteinemia) had LDL with a low receptor affinity, and the degree of the reduction in affinity paralleled the severity of the hypertriglyceridemia. The LDL of hypertriglyceridemic patients was rich in protein and triglycerides, had a low content of cholesterol and phospholipids, and was smaller than normal, thus resembling the atherogenic lipoprotein known as small, dense LDL. These abnormalities were observed even in patients with mild hypertriglyceridemia regardless of their serum cholesterol levels. The degree of alteration in LDL lipid composition and particle size was strongly associated with the reduction of LDL receptor affinity. We also examined the effects of two lipid-lowering agents (bezafibrate and probucol) on the characteristics of LDL. LDL receptor affinity was only improved when the lipid composition and particle size were normalized by drug therapy. Although it has been reported that decreased cholesteryl ester transfer protein (CETP) activity results in the formation of small LDL, plasma CETP activity was normal in the hyperlipoproteinemic patients and the normalization of LDL characteristics by drug therapy was not accompanied by an increase of CETP activity. Our results suggested that an abnormal lipid composition and/or small particle size might cause a decrease in the receptor affinity of LDL. These structural and functional abnormalities were reversed by drug therapy, underlining the importance of treating hypertriglyceridemia for the prevention of atherosclerosis.

Correlation of plasma protein separation patterns obtained by two-dimensional polyacrylamide gel electrophoresis and by capillary electrophoresis.

We used three different electrophoretic techniques for the analysis of human plasma proteins: (i) two-dimensional polyacrylamide gel electrophoresis (2-D PAGE), with sodium dodecyl sulfate (SDS) used only in slab gel electrophoresis; (ii) capillary isoelectric focusing (CIEF) with no denaturants; (iii) linear polyacrylamide (LPA)-filled capillary electrophoresis with SDS (SDS-CE). With technique (i), data on isoelectric point and molecular size of plasma proteins can be obtained. Techniques (ii) and (iii) are suited to obtain quantitative information on proteins. The separation principle used in technique (ii) is closely related to that used in the first dimension of technique (i), and that used in technique (iii) related to that in the second dimension of technique (i). Therefore, we could successfully correlate protein separation patterns obtained by 2-D PAGE and those obtained by capillary electrophoresis. The advantages of correlating data obtained by various electrophoretic techniques in the course of constructing a comprehensive database on human plasma proteins are discussed.

Comparison of the effect of bezafibrate on improvement of atherogenic lipoproteins in Japanese familial combined hyperlipidemic patients with or without impaired glucose tolerance.

The effect of bezafibrate on plasma lipoproteins was investigated in Japanese familial combined hyperlipidemic patients with or without an impaired glucose tolerance accompanied by a low-density lipoprotein subclass, with the major gradient gel peak at a particle diameter of less than 25.5 nm. Bezafibrate treatment at a dose of 400 mg/d for 12 weeks produced an antiatherogenic effect on lipoprotein profiles, as reflected by a decrease in plasma triglyceride levels, an increase in plasma high-density lipoprotein-cholesterol levels, induction of the large-size subclass of low-density lipoprotein, and disappearance of intermediate-density lipoproteins. The plasma total and low-density lipoprotein-cholesterol-lowering effect of bezafibrate was significant in patients without impaired glucose tolerance but was not significant in patients with impaired glucose tolerance. Bezafibrate increased lipoprotein lipase activity and decreased the activity of cholesteryl ester transfer protein, both in patients with or without impaired glucose tolerance. There was no difference in the distribution of signal peptide insertion/deletion or Xbal polymorphisms of the apolipoprotein B gene in patients with or without impaired glucose tolerance. Mechanisms other than lipoprotein lipase, cholesteryl ester transfer protein activities, and an apolipoprotein B gene polymorphism may be responsible for the resistance to lowering of plasma total and low-density lipoprotein cholesterol levels with bezafibrate treatment in familial combined hyperlipidemic patients with impaired glucose tolerance.

Antenatal diagnosis of velamentous umbilical cord insertion and vasa previa with color Doppler imaging.

OBJECTIVE: To determine whether cord insertion can be consistently visualized and whether velamentous cord insertion and vasa previa can be consistently identified with color Doppler imaging during routine sonography in the mid-trimester. DESIGN: A prospective study. SUBJECTS: A total of 587 fetuses at 18-20 weeks' gestation. METHODS: During routine ultrasound examinations, the sonographer was instructed to take additional time and to image the placental cord insertion with color Doppler imaging and classify this as normal, velamentous or 'not seen'. When the insertion was velamentous, the sonographer was instructed to indicate whether or not it was vasa previa. The sonogram obtained at 18-20 weeks' gestation was used for comparison with outcome data. RESULTS: Cord insertion was visualized by color Doppler imaging in 99.8% (586/587) of the fetuses in our study. The mean time required for examination was 20 s and, in 95% of the cases, cord insertion was visualized within 1 min. The sonographic identification of velamentous cord insertion had a sensitivity of 100% (5/5), a specificity of 99.8% (580/581), a positive predictive value of 83% (5/6) and a negative predictive value of 100% (580/580). In our study, vasa previa was diagnosed at 18 gestational weeks in two cases and, in one of the cases, vasa previa was confirmed at delivery. CONCLUSIONS: We could consistently identify cord insertion and velamentous cord insertion with color Doppler imaging during routine sonography in the mid-trimester. Transvaginal color Doppler imaging and serial scans were needed to identify vasa previa.

New method for assaying free and total cholesterol in cultured cells by high-pressure liquid chromatography.

We developed a simple, sensitive and accurate method for assaying cellular free and total cholesterol by monitoring 4-cholesten-3-one, a conversion product of the cholesterol oxidase-catalyzed oxidation of the free cholesterol that has a strong chromophoric alpha, beta-unsaturated ketone at 240 nm, using a high-pressure liquid chromatographic system. This method measured picomole quantities of free and total cholesterol and precisely determined their concentrations in cells (10(4) range) in culture using 7 beta-hydroxycholesterol as an internal standard.

High dose of fluvastatin sodium (XU62-320), a new inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, lowers plasma cholesterol levels in homozygous Watanabe-heritable hyperlipidemic rabbits.

The effects of fluvastatin sodium (XU62-320), a new type of inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A reductase, on plasma cholesterol and triacylglycerol levels were investigated using homozygous Watanabe-heritable hyperlipidemic (WHHL) rabbit, an LDL-receptor-deficient animal which expresses a hepatic LDL receptor activity less than 5% that of control rabbits. Plasma levels of total, VLDL- and LDL-cholesterol were decreased profoundly after oral administration of fluvastatin at a dose of 50 mg/kg per day for 4 weeks. Plasma triacylglycerol levels were not affected by fluvastatin. Hepatic HMG-CoA reductase activity increased by 3-fold and hepatic LDL receptor activity increased by only 3.7-fold, as calculated by Scatchard plot analysis, with fluvastatin administration for 4 weeks, and the hepatic mRNA level for the rabbit LDL receptor was increased by 3-fold. Combined administration of fluvastatin (50 mg/kg per day) and cholestyramine, a bile acid sequestrant resin, at a level of 2% of the diet for 4 weeks more profoundly decreased plasma total, VLDL- and LDL-cholesterol levels with induction of hepatic cholesterol 7 alpha-hydroxylase and no further induction of the hepatic LDL receptor. Plasma triacylglycerol levels were increased by the combination treatment. These results suggest that high dose of fluvastatin sodium is effective in lowering plasma cholesterol levels in homozygous WHHL rabbits through the shared mechanisms involving decrease in production and secretion of cholesterol from the liver and the induction of hepatic LDL receptor. Additional effect of cholestyramine on decrease in plasma cholesterol levels seems to be due to the further decrease in hepatic cholesterol secretion by up-regulation of hepatic cholesterol 7 alpha-hydroxylase.

Different apheresis methods in the treatment of hypercholesterolemia in primary biliary cirrhosis: a case report.

The two different modes of low-density lipoprotein (LDL) apheresis, cascade filtration (CF) and dextran-sulfate cellulose (DSC) adsorption, were compared for efficiency of cholesterol removal in a patient with severe hypercholesterolemia due to primary biliary cirrhosis (PBC). Decrement in the total cholesterol level by the DSC method was less than that by the CF method. Apolipoprotein B was decreased to almost null by both modules whereas the decrease in albumin was much greater in the second filter of the CF method than in the DSC column. Lipoprotein X which constituted the major portion of serum cholesterol in PBC became negative by passing plasma through the second filter. The CF method was preferred to the DSC method for removal of lipoprotein X, but albumin substitution was mandatory to prevent the decrease of serum albumin in the CF method.

Comparison of hypocholesterolemic effects induced by dietary linoleic acid and oleic acid in hamsters.

We investigated the differences between the hypocholesterolemic effects induced by dietary linoleic acid and those induced by oleic acid in hamsters. Addition of 5% linoleic acid or oleic acid to a 0.1% cholesterol-supplemented diet diminished the increases in plasma total and low density lipoprotein (LDL) cholesterol induced by cholesterol alone. Linoleic acid decreased high density lipoprotein (HDL) cholesterol in comparison with cholesterol alone, whereas oleic acid did not. As compared with a standard diet or a cholesterol-supplemented diet, linoleic acid and oleic acid each prevented hepatic LDL receptor suppression, although linoleic acid was more effective. Oleic acid prevented the increase in plasma cholesteryl ester transfer protein (CETP) activity induced by dietary cholesterol, whereas linoleic acid did not. Neither linoleic acid nor oleic acid altered hepatic 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase activity. Only oleic acid increased hepatic cholesterol 7 alpha-hydroxylase activity. These results suggest that dietary linoleic and oleic acids diminish the cholesterol-induced increases in plasma total and LDL-cholesterol by preventing hepatic LDL receptor suppression, and in the case of oleic acid by also preventing the increase in the plasma CETP activity. These effects on cholesterol 7 alpha-hydroxylase activity may influence bile lipid metabolism.

[Acrodystrophic neuropathy in an alcoholic].

The patient was a 48-year-old alcoholic man with no contributory family history. At age 36 he had developed sensory dominant polyneuropathy with highly impaired temperature sensation and deep sensation in the lower extremities, recurrent ulcers of the toes, and sexual impotence. A sural nerve biopsy at this time revealed marked loss of myelinated fibers with relative preservation of the population of unmyelinated fibers. Subsequently, he developed muscle atrophy of the lower thighs, urinary incontinence, and Wernicke's encephalopathy, and became non-ambulatory at age 44. The peripheral nerve conduction findings suggested predominantly axonal degeneration. The entire course was characterized by alternative progression and partial recovery influenced by his alcohol intake and nutritional state. Alcoholic neuropathy is a major cause of solitary acrodystrophic neuropathy (ADN). Manifestations of autonomic and motor neuropathy are more marked in alcoholic ADN than in HSAN-I, and central nervous system involvement is the hallmark of alcoholic ADN. In the treatment of patients with alcoholic ADN, attention should be paid to diabetes mellitus, malnutritional state, and vitamin deficiency, which frequently complicate alcoholism.

Changes in coagulation factors by passage through a dextran sulfate cellulose column during low-density lipoprotein apheresis.

To determine the extent of adsorption of coagulation factors by a dextran sulfate cellulose column used for selective removal of low-density lipoprotein (LDL), various coagulation factors were measured before and after application to the column during LDL apheresis. The column almost completely adsorbed many coagulation factors. Although the bradykinin concentration was markedly increased by passing the plasma through the column, this increment was suppressed by nafamostat mesilate which inhibits the initial contact phase of the intrinsic coagulation pathway. The von Willebrand factor, which forms a complex with factor VIII in plasma, is reduced in apheresis with nafamostat mesilate to the same extent as in apheresis without nafamostat mesilate. Thus, coagulation factors seem to be adsorbed by different mechanisms which include activation of the initial contact phase by the negative charges of dextran sulfate and concomitant adsorption with the phospholipid portion of lipoproteins containing apolipoprotein B or with von Willebrand factor.