A new disorder of praxis in neurodegenerative disease that may be part of Alzheimer's disease.

Apraxia is a well-known disorder of praxis and is caused mainly by damage to the left parietal lobe. We presented two cases of neurodegenerative disease with a distinct disorder of praxis, predominantly involving left parietal lobe. While both patients could understand what they should do, they were not able to initiate action and often stopped during execution of actions. They had no apraxia and no temporal and spatial errors on praxis. Magnetic resonance imaging of both patients showed atrophy of the left parieto-occipital and temporo-occipital lobes, and single photon emission computed tomography showed hypoperfusion in the same lobes. Moreover, one of our cases, using [11C] PIB PET, demonstrated increased uptake in the cerebral cortices, suggesting Alzheimer's disease. The symptoms described are different from other disorders of praxis and similar to bradyphrenia or freezing.

Relationship between response of gamma-glutamyl transpeptidase to alcohol drinking and risk factors for coronary heart disease.

Alcohol drinking has been reported to influence the risk factors for coronary heart disease (CHD), such as the serum levels of triglycerides, HDL-cholesterol and uric acid, and the level of blood pressure. To examine whether there was individual variability in the response of these parameters to alcohol drinking, a cross-sectional study was conducted in 3130 men with a body-mass index (BMI) below 24. The subjects were divided into two groups; a normal gamma-glutamyl transpeptidase (rGTP) (<40 IU/l) group and a high rGTP (> or =40 IU/l) group, and the values were compared after adjusted for age, BMI, exercise and smoking. The level of triglycerides increased according to the amount of drinking in the high rGTP group, whereas no association was observed in the normal rGTP group. The level of HDL-cholesterol increased with drinking in the normal and high rGTP groups, and no difference was observed in the levels of HDL-cholesterol between the two groups. The levels of uric acid and blood pressure also increased with drinking in both groups, but the increase was bigger in the high rGTP group than in the normal rGTP group. The results indicated that there was large individual variability in the responses of the risk factors for coronary heart disease to drinking. Subjects whose rGTP responds less to drinking may have less disadvantageous effects of drinking.

[Reference intervals of urinary biochemical analysis in Japanese men].

To determine the reference intervals of urinary albumin, immunoglobulin G, transferrin, alpha 1 microglobulin, and N-acetyl-beta-D-glucosaminidase, we measured those components and creatinine in urine from men with normal blood pressure(systolic < or = 139 mmHg and diastolic < or = 89 mmHg), HbA1c < or = 5.8% and negative qualitative tests of urinary protein and hematuria. The subjects were 150 men in their thirties, 136 in their forties, and 135 in their fifties. Urines was collected in the morning after at least 12 hours fast and stored at -80 degrees C until assayed. The levels of creatinine decreased with age, whereas no change was observed in the levels of other urinary components. Consequently, the levels of those components expressed as g. creatinine increased with age. The reference intervals per g. creatinine were determined for each age-bracket using the iterative truncation method after logarithmical transformation of the values.

Conformational equilibrium of the reactive center loop of antithrombin examined by steady state and time-resolved fluorescence measurements: consequences for the mechanism of factor Xa inhibition by antithrombin-heparin complexes.

Activation of antithrombin by high-affinity heparin as an inhibitor of factor Xa has been ascribed to an allosteric switch between two conformations of the reactive center loop. However, we have previously shown that other, weaker binding, charged polysaccharides can give intermediate degrees of activation [Gettins, P. G. W., et al. (1993) Biochemistry 32, 8385-8389]. To examine whether such intermediate activation results from different reactive center loop conformations or, more simply, from a different equilibrium constant between the same two extreme conformations, we have used NBD covalently bound at the P1 position of an engineered R393C variant of antithrombin as a fluorescent reporter group and measured fluorescence lifetimes of the label in free antithrombin as well as in antithrombin saturated with long-chain high-affinity heparin, high-affinity heparin pentasaccharide, long-chain low-affinity heparin, and dextran sulfate. Steady state emission spectra, anisotropies, and dynamic quenching measurements were also recorded. We found that the large steady state fluorescence enhancements produced by binding of activators resulted from relief of a static quench of fluorescence of NBD in approximately 50% of the labeled antithrombin molecules rather than from any large change in lifetimes, and that similar lifetimes were found for NBD in all activated antithrombin-oligosaccharide complexes. Similar anisotropies and positions of the NBD emission maxima were also found in the absence and presence of activators. In addition, NBD was accessible to quenching agents in both the absence and presence of activators, with an at most 2-fold increase in quenching constants between these two extremes. The simplest interpretation of the partial static quench in the absence of activators, the different degrees of enhancement by different antithrombin activators, and the similar fluorescence properties and quenching behavior of the different states is that there are two distinct types of conformational equilibrium involving three distinct states of antithrombin, which we designate A, A', and B. A and A' represent low-affinity or inactive states of approximately equal energy, both having the hinge residues inserted into beta-sheet A. A is fluorescent, while A' is statically quenched. State B represents the activated loop-expelled conformation in which none of the NBD fluorophores are statically quenched, as a result of the loop, including the P1-NBD, moving away from the body of the antithrombin. Different activators are able to shift the equilibrium to the high-activity (B) state to different extents and hence give different degrees of measured activity, and different degrees of relief of static quench. The similar properties and accessibility of the NBD in the A and B conformations also indicate that the P1 side chain is not buried in the low-activity A conformation, suggesting that an earlier proposal that activation involves exposure of the P1 side chain cannot be the explanation for activation. As an alternative explanation, heparin activation may give access to an exosite on antithrombin for binding to factor Xa and hence be the principal basis for enhancement of the rate of inhibition.

Effects of the frequency of alcohol intake on risk factors for coronary heart disease.

A cross-sectional study was conducted in 3660 male workers to examine whether the weekly frequency of alcohol intake affected serum lipids and blood pressure, which are risk factors for coronary heart disease, independently of the weekly alcohol consumption. Information regarding life-style habits and current medication was obtained by questionnaire. The effects of the frequency were examined using Tukey's test in the groups of drinkers divided according to their alcohol consumption. In moderate (189-377 ml/week) drinkers, a higher frequency of drinking was related to a higher level of HDL-cholesterol, and a lower level of triglycerides. In light (1-188 ml/week) and heavy (378-566 ml/week) drinkers, a higher frequency of drinking was also related to a higher level of HDL-cholesterol. There were no significant relations between the frequency of drinking and total cholesterol, or blood pressure in these three groups. Similar results were obtained when values were adjusted for age, body-mass index, smoking, physical activity and weekly alcohol consumption. Multiple regression analysis in the whole drinkers also showed that the weekly frequency of drinking was associated with HDL-cholesterol, but not with triglycerides (p = 0.052), total cholesterol, or blood pressure. The results suggest that the weekly frequency of drinking may affect the levels of HDL-cholesterol independently of the weekly alcohol consumption.

Secondary tethered cord syndrome in spinal dysraphism.

Secondary tethered cord syndrome following initial repair for spinal dysraphism is an important area of interest. In this study, 32 cases with spinal dysraphism in the lumbosacral region were enrolled, in whom radical repair with autologous material had been carried out in the early stage soon after birth. During the follow-up period of up to 19 years 10 months, surgery was considered to be indicated in 2 of the 8 lipomeningocele cases and in 6 of the 24 meningocele and meningomyelocele cases, because of the presence of tethered cord syndrome 4-19 years after the primary operation. In all 8 of these cases, MR imaging demonstrated tethered spinal cord in the form of low conus medullaris. In 6 of the 8 operated cases surgery was followed by improvement of the spinal neurological deterioration. According to our experience, early untethering for secondary tethered cord syndrome is essential. In addition, since the complications of Silastic duraplasty at untethering were all minor and the operative outcome was satisfactory, the use of silicone rubber sheeting as a dural substitute might be recommended to prevent adhesion of the spinal cord.

Serine 380 (P14) --> glutamate mutation activates antithrombin as an inhibitor of factor Xa.

Heparin regulates the inhibitory activity of antithrombin. It has been proposed that residues P15 and P14 are expelled from beta-sheet A of antithrombin by heparin binding, permitting better interaction of the reactive center loop with factor Xa. We have made a P14 antithrombin variant (S380E) to create an activated inhibitory form of antithrombin in which P14 is already expelled from beta-sheet A. S380E antithrombin fluorescence is enhanced 35 +/- 5% compared with control antithrombin. There is minimal further increase in antithrombin fluorescence upon heparin binding. The variant has a 5 degrees C lower T(m) than control antithrombin. The variant is an inhibitor of proteinases and has a nearly 200-fold increased basal rate of inhibition of factor Xa, after correction for an increased stoichiometry of inhibition. This is comparable to that of antithrombin activated by high affinity heparin pentasaccharide. Full-length high affinity heparin causes only a 7-fold additional increase in rate and a large increase in stoichiometry of inhibition. In contrast, the basal rate of inhibition of thrombin is similar to that of control antithrombin but is increased 300-fold by heparin. These findings suggest that the native state of the S380E variant exists in a loop-expelled conformation that is consequently highly reactive toward factor Xa.

Relationship between the serum level of leptin and life-style habits in Japanese men.

AIMS/METHODS: To investigate whether smoking affects the serum level of leptin, 708 male workers aged 25-65 years old were cross-sectionally surveyed. RESULTS: Multiple regression analysis indicated that among the various parameters examined, the level of leptin was positively associated with the body mass index and the levels of insulin, total cholesterol and uric acid, and was inversely associated with physical activity and the level of creatinine. The partial correlation coefficient of leptin was highest against the body mass index (r = 0.40), followed by insulin (0.29) and physical activity (-0.14), after adjustment for other leptin-related variables. However, no association was observed between the level of leptin and smoking (0.05), alcohol consumption (0.09) or age (0.09). CONCLUSIONS: The findings suggest that among life-style habits, physical activity, but not smoking or alcohol consumption, significantly affects the serum level of leptin in Japanese men.

Two different types of dehalogenases, LinA and LinB, involved in gamma-hexachlorocyclohexane degradation in Sphingomonas paucimobilis UT26 are localized in the periplasmic space without molecular processing.

gamma-Hexachlorocyclohexane (gamma-HCH) is one of several highly chlorinated insecticides that cause serious environmental problems. The cellular proteins of a gamma-HCH-degrading bacterium, Sphingomonas paucimobilis UT26, were fractionated into periplasmic, cytosolic, and membrane fractions after osmotic shock. Most of two different types of dehalogenase, LinA (gamma-hexachlorocyclohexane dehydrochlorinase) and LinB (1,3,4,6-tetrachloro-1,4-cyclohexadiene halidohydrolase), that are involved in the early steps of gamma-HCH degradation in UT26 was detected in the periplasmic fraction and had not undertaken molecular processing. Furthermore, immunoelectron microscopy clearly showed that LinA and LinB are periplasmic proteins. LinA and LinB both lack a typical signal sequence for export, so they may be secreted into the periplasmic space via a hitherto unknown mechanism.

[Effects of a 5-day fast on clinical laboratory data from patients with rheumatoid arthritis].

There have been few studies on the effects of a fast on clinical laboratory data in Japanese. We studied twelve women with rheumatoid arthritis who were not taking any medicine and stayed in the Koda hospital for a diet which lasted 55 days. They basically took a 1200 kcal vegan diet and undertook a 3-5-day fast three times. The clinical laboratory data obtained before and after the second fast (day 27-day 31) were compared. Average body weight decreased by 1.5 kg. There were no changes in CRP. Rapid turnover proteins such as alpha 1 and beta 2-microglobulin decreased, whereas albumin, IgG, IgA and IgM increased. HDL-C increased without a change in LDL-C or triglycerides. Free T3 decreased and free T4 increased, while TSH did not change. The increases in albumin, Ig, HDL-C and free T4 were not consistent with the results of previous studies. This difference may have been due to the low calorie vegan diet before the fast.

[Urine C-peptide excretion in hypocaloric states and factors affecting its excretion].

Recent evidence suggests that hyperinsulinemia may contribute to the development of various risk factors of atherosclerosis. To examine the effects of energy intake on insulin secretion, 24-h urine C-peptide was measured in twelve women with rheumatoid arthritis who were not taking any medicine and stayed in Koda hospital for a diet therapy which lasted 55 days. They were basically placed on a 1200 kcal/day vegan diet combined with three 3-5-day fasting periods (200 kcal/day). Urine C-peptide excretion markedly decreased from 31-40 to 8-14 micrograms/day during the fasting periods. Among the anthropometric variables examined, the average level of urine C-peptide excretions measured in the fasting periods showed a significant correlation with the percentage and the amount of body fat. However, such correlation was not observed while the calorie intake was 1200 kcal. No clinical laboratory parameter showed a significant correlation with urinary C-peptide excretion. These results suggest that the major determinant of urine C-peptide excretion is food intake and that hyperinsulinemia could be easily improved by restricting energy intake.

[Relationship between the prevalence of proteinuria and obesity in Japanese men].

5174 Japanese male workers were studied cross-sectionally to examine a possible relationship between obesity and the prevalence of proteinuria (1 + or greater), determined using a reagent strip. The subjects were divided into three groups with body-mass indexes (BMIs) of < 23, 23-24.9 and > or = 25, and the prevalence of proteinuria was compared among the three groups, after adjustments for age, blood pressure and blood levels of HbA1c. Among subjects with hypertension and/or a high level of HbA1c, i.e., > or = 5.9%, the group with a BMI of > or = 25 had a higher prevalence of proteinuria than the group with a BMI of < 23. In contrast, the prevalence of proteinuria was not affected by obesity in the subjects with neither hypertension nor a high level of HbA1c. These results suggest that for obese men with hypertension or diabetes, reducing body weight is important not only as part of the treatment for those diseases, but also to prevent proteinuria.

[Comparison of LDL-C values measured with the automated method and the ultracentrifugation method in severe hypertriglyceridemia, and prevalence and life-style of patients with hypertriglyceridemia].

The correlation between LDL-cholesterol (LDL-C) values assayed by the direct method and the ultra-centrifugation method is reported good in normal to moderate hypertriglyceridemia, but it is not clear in severe hypertriglyceridemia. We examined such a correlation in mild (triglycerides, 150-400 mg/dl; n = 3) and severe (> or = 800 mg/dl, n = 9) hypertriglyceridemia. The bias of LDL-C determined by the direct method in comparison with the ultracentrifugation method was from -1.1% to 3.4% and from -49.5% to 15.7% in mild and severe hypertriglyceridemia, respectively. The prevalence of severe hypertriglyceridemia was only 0.2% both in hospital patients and in company workers. Data analyses of company workers indicated that people with severe hypertriglyceridemia have a higher body-mass index, consume more alcohol, smoke more, and exercise less than those with a normal level of triglycerides. These results suggest that there is not a good correlation between LDL-C values assayed by the direct method and the ultracentrifugation method in severe hypertriglyceridemia; but that the direct method can be used for the clinical examination of LDL-C, because of the very low prevalence of severe hypertriglyceridemia. Patients with severe hypertriglyceridemia should improve their life-style as soon as possible.

Effect of the platelet-derived growth factor antagonist trapidil on mesangial cell proliferation in rats.

Platelet-derived growth factor (PDGF) is known as a potent mediator in the proliferation of mesangial cells in culture and in mesangial proliferative nephritis. The present study was undertaken to evaluate the effect of trapidil, an antagonist of PDGF, on mesangial cell proliferation in culture and in anti-Thy-1.1 nephritis in rats. Trapidil significantly inhibited 3H-thymidine incorporation in the mesangial cells stimulated by PDGF BB and suppressed mesangial cell proliferation in culture in a dose-dependent manner. In anti-Thy-1.1 nephritis, a significant reduction in the number of total glomerular cells and also proliferating (proliferating cell nuclear antigen positive) cells was demonstrated on day 7 in the rats treated with trapidil as compared with controls. Although renal function expressed as blood urea nitrogen and creatinine levels did not differ between rats with and without trapidil treatment, the present results suggest a salutary effect of trapidil on mesangial cell proliferation. PDGF, therefore, could play an important role in mediating mesangial cell proliferation.

Relationship between glycosylated hemoglobin and the prevalence of proteinuria in Japanese men.

A total of 5,174 Japanese men were included in a cross-sectional study to examine the relationship between the glycated hemoglobin (HbA1C) level and the prevalence of proteinuria as determined using a reagent strip. The prevalence of proteinuria rose significantly at HbA1C levels above 5.9%, whereas no relationship was observed at HbA1C levels below 5.9%. Multiple logistic regression analysis showed that blood pressure and a family history of diabetes were independent factors associated with proteinuria in subjects with a HbA1C below 5.9% who were not under medication for diabetes. In contrast, HbA1C, obesity and smoking were associated with proteinuria in subjects who were under medication for diabetes and/or have a HbA1C above 5.9%. These findings suggest that maintaining a HbA1C level below 5.9%, non-smoking and a standard body weight may reduce the prevalence of proteinuria in Japanese men. Healthy life-style and standard body weight are especially important for subjects with a family history of diabetes.

A rare case of sarcoidosis with bilateral leg lymphedema as an initial symptom.

A 55-year-old man was admitted to our hospital because of bilateral leg lymphedema. He also showed subcutaneous nodules and CT scan disclosed multiple enlarged lymph nodes at thoracic, abdominal, and inguinal areas. Biopsy of the inguinal lymph node and the subcutaneous nodule revealed noncaseating epithelioid cell granuloma, a finding consistent with sarcoidosis. Lymphedema was attributed to the blockade of lymph flow by the systemic lymph node involvement of the disease. Within 1 week after the initiation of steroid therapy, his leg edema disappeared. Lymphedema could be the initial symptom of systemic sarcoidosis.

Change in environment of the P1 side chain upon progression from the Michaelis complex to the covalent serpin-proteinase complex.

Serpins inhibit proteinases by forming a kinetically trapped intermediate during a suicide substrate inhibition reaction. To determine whether the kinetic trap involves a repositioning of the P1 side chain of the serpin following formation of the initial Michaelis complex, we used the tryptophan of a P1 M-->W variant of human alpha1-proteinase inhibitor as a fluorescent reporter group of the environment of the P1 side chain. The P1W variant was a valid model serpin and formed SDS-stable complexes with both trypsin and chymotrypsin with a stoichiometry of inhibition close to 1.0. Rates of inhibition of chymotrypsin for wild-type and variant alpha1-proteinase inhibitor differred only approximately 1.8-fold. Rates of inhibition of trypsin were, however, 25-fold lower for the variant than for the wild-type inhibitor. Steady-state fluorescence spectra showed a change in environment for the P1 side chain upon forming both covalent complex with trypsin or chymotrypsin and noncovalent complex with anhydrochymotrypsin. The P1 environments in the chymotrypsin and anhydrochymotrypsin complexes were, however, different. Fluorescence quenching studies confirmed the burial of the P1 side chain upon formation of both the noncovalent and covalent complexes, but were not able to discriminate between the solvent accessibility in these complexes. Stopped-flow fluorescence measurements resolved the covalent intramolecular reaction that led to covalent complex and showed that, during the course of the covalent reaction, the environment of the P1 side chain changed consistent with a repositioning relative to residues of the proteinase active site as part of formation of the trap. This repositioning is likely to be a crucial part of the trapping mechanism.

[Adult-onset type II citrullinemia in a patient undergoing continuous ambulatory peritoneal dialysis].

A 57-year-old Japanese man was admitted to Toyama Medical & Pharmaceutical University Hospital with delirium and flapping tremor on April 2, 1997. He had been undergoing continuous ambulatory peritoneal dialysis (CAPD) because of diabetic nephropathy since 1993. Blood chemistry showed slightly elevated plasma ammonia level with no evidence of liver injury, and his portal venography revealed no port-systemic shunt. He was diagnosed as having type II citrullinemia because of an elevated citrulline level on amino acid analysis and very low hepatic argininosuccinate synthetase activity obtained from biopsy specimen of liver. In this case, plasma concentrations of ammonia and citrulline were not so high as those in previously reported cases, although the hepatic argininosuccinate synthetase activity was actually less than 10% of the control value. Owing to CAPD, he was conservatively controlled in a relatively good condition. This indicates that CAPD seems to be a useful therapeutic approach for citrullinemia since liver transplantation is still difficult in Japan.