Multiple optical stimulation to neuron using Si opto-neural probe with multiple optical waveguides and metal-cover for optogenetics.

We have developed a Si opt-neural probe with multiple waveguides and metal cover for highly accurate optical stimulation. This neural probe had 16 recording sites, three optical waveguides, and metal cover for suppressing light leakage. We evaluated electrochemical properties of the recording sites, and confirmed that the neural probe had suitable characteristics for neural recording. We also demonstrated the optical stimulation to the neurons expressing ChR2 using our probe. As a result, we succeeded multisite optical stimulation, and observed that no light leakage from the optical waveguides because of the metal cover. From in vivo experiments, we successfully recorded optically modulated local field potential using the fabricated Si neural probe with optical waveguides. Moreover, we applied current source density analysis to the recorded LFPs. As a result, we confirmed that light induced membrane current sink in locally stimulated area. Our Si opto-neural probe with multiple optical waveguides and metal-cover is one of the most versatile tools for optogenetics.

Seroprevalence of toxoplasmosis among migrant workers from different Asian countries working in Malaysia.

A serologic study of Toxoplasma antibodies among 501 foreign migrant workers in Malaysia was conducted in a plantation and detention camp. The highest prevalence rate of 46.2% was among Nepalese workers. Statistical analysis indicated the IgG positivity rate among local residents was significantly higher than the migrants studied (p < 0.05). The IgM positivity rate showed no significant difference between the two groups (p > 0.05). No significant difference in the prevalence rate was noted between the migrants and the local workers when grouped by agricultural and non-agricultural occupations (p > 0.05). The continuous introduction of these infections may influence the epidemiology and further compromise efforts in control and prevention. It is therefore important to monitor of non-notifiable diseases.

Variationally optimized basis orbitals for biological molecules.

Numerical atomic basis orbitals are variationally optimized for biological molecules such as proteins, polysaccharides, and deoxyribonucleic acid within a density functional theory. Based on a statistical treatment of results of a fully variational optimization of basis orbitals (full optimized basis orbitals) for 43 biological model molecules, simple sets of preoptimized basis orbitals classified under the local chemical environment (simple preoptimized basis orbitals) are constructed for hydrogen, carbon, nitrogen, oxygen, phosphorous, and sulfur atoms, each of which contains double valence plus polarization basis function. For a wide variety of molecules we show that the simple preoptimized orbitals provide well convergent energy and physical quantities comparable to those calculated by the full optimized orbitals, which demonstrates that the simple preoptimized orbitals possess substantial transferability for biological molecules.

Role of a highly conserved YPITP motif in 2-oxoacid:ferredoxin oxidoreductase: heterologous expression of the gene from Sulfolobus sp.strain 7, and characterization of the recombinant and variant enzymes.

2-Oxoacid:ferredoxin oxidoreductase from Sulfolobus sp. strain 7, an aerobic and thermoacidophilic crenoarchaeon, catalyses the coenzyme A-dependent oxidative decarboxylation of pyruvate and 2-oxoglutarate, a cognate Zn-7Fe-ferredoxin serving as an electron acceptor. It comprises two subunits, a (632 amino acids) and b (305 amino acids). To further elucidate its structure and function, we constructed a gene expression system. The wild-type recombinant enzyme was indistinguishable from the natural one in every criterion investigated. A series of variants was constructed to elucidate the role of the YPITP-motif (residues 253-257) in subunit a, which is conserved universally in the 2-oxoacid:ferredoxin oxidoreductase (OFOR) family. Single amino-acid replacements at Y253 and P257 by other amino acids caused a drastic loss of enzyme activity. T256, the hydroxyl group of which has been proposed to be essential for binding of the 2-oxo group of the substrate in the Desulfovibrio africanus enzyme, was unexpectedly replaceable with Ala, the kcat and Km for 2-oxoglutarate being approximately 33% and approximately 51%, respectively, as compared with that of the wild-type enzyme. Replacement at other positions resulted in a significant decrease in the kcat of the reaction while the Km for 2-oxoacid was only slightly affected. Thus, the YPITP-motif is essential for the turnover of the reaction rather than the affinity toward 2-oxoacid.

A brief survey of free-living amebae in Thailand and Hamamatsu District, Japan.

The aim of this study was to determine the presence of free-living amebae in aquatic habitats of human environments in Thailand and Hamamatsu district, Japan. Genus identification was based on the morphology of cyst and trophozoite forms and a flagellation test for genus Naegleria. The pathogenic potential was tested in mice by nasal instillation for genus Naegleria and Acanthameba. In 14 provinces of Thailand, amebae were isolated in 43 from 95 water samples and 67 from 120 soil swabs. Amebae of 49 isolates from waters were identified as Acanthameba (36.7%), Naegleria (28.6%), Hartmannella (20.4%), Vahlkampfia (12.2%) and Vannella (2%). Soil samples have significantly higher levels of Acanthameba and Hartmannella (p<0.05) but lower for Naegleria (p<0.05) and 7 unidentified amebae were found. In Hamamatsu district, Japan, 62 amebae of the same genera were isolated from 47 of 95 water samples. There were significantly higher levels of Acanthameba (22.6%) (p<0.05) but lower for Naegleria (4.8%) (p<0.05) than those of Thailand which each of them caused death in mice. Three unidentified amebae were isolated. This finding serves as additional evidence for the presence of free-living amebae under natural and the difference in distribution between tropic and subtropic areas.

Epidemiology, symptoms and treatment of paragonimiasis in Sin Ho district, Lai Chau province, Vietnam.

Sin Ho is a district of the northern mountainous province of Lai Chau, Vietnam, where the people have the habit of eating undercooked crabs. A study on paragoniamiasis carried out from 1994 to 1995 with 1,642 persons in this endemic area, showed that the rate of eating raw-crab was 72.5%. Crab examination (Ranguna kimboiensis) showed an infection rate of Paragonimus metacercaria of 98.1%. With 624 stool samples examined by Kato technique, the infection rate of Paragonimus in humans was 6.4%. With 338 sputum samples examined by direct and centrifuge methods, the infection rate of Paragonimus was 7.4%. Most of the patients were children (63.2%). The infection rate of Paragonimus in dogs was 18.2 - 33.3%. Adult worms, collected from the dogs in the field and from the cats in laboratory, were identified as Paragonimus heterotremus. The main symptoms of Paragonimus patients were cough and hemoptysis (92%), discontinuously developed (96%), without fever (94%), chest pain (70%), pleural effusion (26%), neurogical symptoms (8%), eosinophilia (88.9%), nodular ring shadows in the lungs, as shown by chest X-ray examination and more in lower lobe, (76.2%). Paragonimiasis patients were treated by (a) Praziquantel 25 mg/kg/day x 3 days; the cure rate was 68.8%. (b) Praziquantel 50 mg/kg/day x 3 days; the cure rate was 75%.

Significant role of the increase in renin-angiotensin system in cardiac hypertrophy and renal glomerular sclerosis in double transgenic tsukuba hypertensive mice carrying both human renin and angiotensinogen genes.

Tsukuba hypertensive mice (THM) are a hypertensive model prepared by mating a transgenic mice with human renin gene and a transgenic mice with human angiotensinogen gene. In the present study, we examined effects of renin-angiotensin system (RAS) on cardiac hypertrophy and renal disorders using Tsukuba hypertensive mice. While THM showed an increase of about 30 mmHg in systolic pressure compared to C57BL/6 mice employed as normal control animals, the increase in blood pressure was not observed in the mice to which either gene was transferred. Urinary volume, water intake volume, urinary albumin excretion, heart to body weight ratio and renal glomerular sclerosis index increased significantly in THM, but none of these parameters showed a significant difference from the C57 mice when they were examined in mice to which either of the genes was transferred. In contrast, when lisinopril was administered to THM, all the parameters decreased significantly without lowering the systolic pressure. From these findings, it was demonstrated that RAS was playing a significant role in cardiac hypertrophy and renal disorders of THM and that lisinopril had inhibitory effects on cardiac hypertrophy and renal glomerular sclerosis by inhibiting RAS.

Role of the renin-angiotensin system in cardiac hypertrophy and renal glomerular sclerosis in transgenic hypertensive mice carrying both human renin and angiotensinogen genes.

Tsukuba hypertensive mice (THMs) are transgenic mice carrying human renin and angiotensinogen genes. The aim of this study was to evaluate the role of the renin-angiotensin system (RAS) in cardiac hypertrophy and renal disorders in THMs. After a 2-wk control period, 10-wk-old THMs were treated with lisinopril (ACEI group) or hydralazine (hydralazine group) or left untreated (control group) for 8 wk. C57BL/6 mice of similar age (wild group) were used as normal controls. Systolic blood pressure and urinary albumin excretion were measured once a week. All mice were sacrificed at 20 wk of age, and heart to body weight ratio, cardiac myocyte diameter, renal glomerular sclerosis index, and glomerular size were measured. Fibronectin expression was also evaluated. At 20 wk of age, systolic blood pressure and urinary albumin excretion in the control group were significantly higher than those in the wild group and significantly lower than those in the ACEI and hydralazine groups. Heart to body weight ratio and cardiac myocyte diameter were significantly higher in the hydralazine and control groups than in the other groups. Renal glomerular sclerosis index and glomerular size were also significantly higher in the control group than in the other groups, and there were significant differences between the ACEI and hydralazine groups in these variables. Fibronectin expression was marked in the control and hydralazine groups. These findings suggest that the RAS plays an important role in cardiac hypertrophy in THMs, but that both the RAS and elevation of blood pressure contribute to the pathogenesis of renal glomerular sclerosis.

Epidemiology of clonorchiasis in Ninh Binh Province, Vietnam.

Clinical and stool examinations for clonorchiasis were carried out in an endemic area, Kim Son District, Ninh Binh Province, Vietnam. Stool examination with the Kato-Katz technic revealed that in 306 residents selected randomly, 42 people (13.7%) were infected with Clonorchis sinensis. The rate was biased towards men (23.4%) as opposed to women (1.5%) and increased with age. No children younger than 10 years old were infected, reflecting difference in a chance for acquisition of infection through a habit of eating raw fish. Few clinical abnormalities were found by blood and urine examinations of the patients. Treatment with praziquantel decreased the infection rate to 5.3% at 6 weeks later. Snails, Melanoides tuberculatus, collected from ponds around the settlements were infected with cercariae at a rate of 13.3%. Farmed fish (Hypophthalmichtys molitrix) in the ponds were infected with metacercariae at rates of 56.4% in small individuals and 100% in large ones. The life cycle of C. sinensis is exclusively completed in the ponds and the traditional habit of eating raw fish in summer was thought to be a major route of infection.

[Medical practice-supporting system in pulmonary function testing].

A pulmonary function testing system which automates measurement, storage and reporting as much possible has been functioning in Mitsui Memorial Hospital since 1992. Data obtained from four measurement apparatus are transferred to the central microcomputer by a local area network (LAN). To facilitate automatic interpretation, we developed a knowledge-based system using 87 production rules called 'PADRES', because the knowledge-based system had the following advantages over other systems; (1) The expression of knowledge is understandable and new pieces of knowledge can be easily added, (2) the process of reasoning can be clearly shown, and (3) the accumulation of many instances of data is not required. Using 'PADRES', it takes 2.6 seconds to interpret one set of data on average. Our system performs about 5,000 routine pulmonary function tests per year without faults. Because the program for interpretation is a separate module written in C language, it is readily portable to other operating systems (OS), although its original target was OS-9, a multitask operating system for microcomputers. In conclusion, the application of a knowledge-based system for microcomputers in the pulmonary function test laboratory was considered successful.

Atrial natriuretic peptide and antihypertensive action due to beta-blockade in essential hypertensive patients.

The effects of beta-blocker treatment on hemodynamics were studied in relation to plasma atrial natriuretic peptide (ANP) levels in 17 outpatients with essential hypertension. Administration of propranolol for twelve weeks to untreated subjects resulted in a significant (P < 0.001) rise in plasma ANP levels (from 37.9 +/- 21.2 to 66.7 +/- 46.2 pg/mL, mean +/- SD). Systolic and diastolic blood pressures were significantly decreased (P < 0.05 and P < 0.01, respectively). Heart rate was also significantly decreased (P < 0.001). On the other hand, a significant reduction of cardiac index was detected (from 4.12 +/- 1.34 to 2.96 +/- 0.75 L/min/m2, P < 0.01) with chronic administration of propranolol, suggesting a reflection of decreased cardiac function. A significant negative correlation was observed between %changes in systolic blood pressure and %changes in plasma ANP (r = -0.594, P < 0.05). These results suggest that the increased plasma ANP levels may contribute to the antihypertensive effect with propranolol.

Effect of an angiotensin II receptor antagonist, TCV-116, on neointimal formation following balloon injury in the SHR carotid artery.

1. In the present study, we examined the effect of a novel angiotensin II type I receptor antagonist, TCV-116, on carotid neointimal formation after balloon injury in SHR and WKY rats. 2. Oral administration of TCV-116 at a dose of 10 mg/kg per day reduced not only systolic blood pressure but also neointimal formation after carotid balloon injury. TCV-116 also suppressed cardiac hypertrophy. An angiotensin-converting enzyme inhibitor, lisinopril (20 mg/kg per day), had a similar effect to that of TCV-116. 3. In the WKY experiment, both TCV-116 and lisinopril suppressed neointimal formation as well as systolic blood pressure, but did not suppress cardiac hypertrophy. 4. Although SHR showed markedly enhanced neointimal formation after balloon injury compared with age-matched WKY rat, both TCV-116 and lisinopril showed similar suppressive effects on neointimal formation in both SHR and WKY rats. 5. These results confirm the important role of angiotensin II in neointimal formation following balloon injury. Further studies are needed to clarify the mechanism of the difference between SHR and WKY rats in the response of vascular smooth muscle cells.

Pathologic changes related to subcutaneous implantation of chlormadinone acetate for preventing estrus in bitches.

Pathologic changes related to chlormadinone acetate (CAP) implantation were examined using 14 bitches given doses 2.5 to 25 mg/kg for 2 years. Absence of corpus luteum in bitches given 5 mg/kg or more supported long-term preventive effect of CAP on estrus. The uteri were dose-dependently enlarged and mucometra was occasionally found. Endometrial epithelium hyperplasia was observed but less in smaller doses. Changes in the mammary gland were only growth and lactation at normal degree. No remarkable changes were observed in ACTH and LH cells in the pituitary gland. Low, stable levels of CAP maintained in plasma by subcutaneous implantation seemed to be the main reason for absence or slight CAP-related pathologic changes.

Effect of an angiotensin II receptor antagonist, TCV-116, on rat carotid artery neointimal formation after balloon injury.

Arterial injury by a balloon catheter produces marked smooth muscle cell proliferation and the participation of angiotensin II in this response has been suggested. In this study, we examined the effect of a novel angiotensin II type I receptor antagonist, TCV-116, on neointimal formation after rat carotid artery balloon injury. Oral administration of TCV-116 at doses of 1, 5 or 10 mg/kg/day significantly reduced the cross-sectional intimal area by 30%, 46% and 54%, respectively, and reduced the ratio of the intimal to medial cross-sectional areas by 23%, 41% and 50%. An angiotensin-converting enzyme inhibitor, lisinopril, had an effect similar to that of TCV-116. The effect of both drugs was significantly correlated with the reduction of both blood pressure and cardiac hypertrophy. We conclude that TCV-116 can prevent neointimal formation after balloon injury as well as reducing blood pressure and preventing cardiac hypertrophy.

Characterization of renal aldosterone receptors in genetically hypertensive rats.

To investigate the aldosterone responsiveness of genetically hypertensive rats, we compared characteristics of renal cytosolic aldosterone receptors from the M strain of stroke-prone, spontaneously hypertensive rats (M-SHRSP) with normotensive Wistar-Kyoto rats (WKY). In M-SHRSP, blood pressure was elevated significantly at 6 wk of age, when their plasma aldosterone concentrations were similar to those in WKY. Decreases in urine volume and sodium excretion were also observed in M-SHRSP. At 10 wk of age, M-SHRSP plasma aldosterone concentrations became significantly higher than those in WKY. On the other hand, the concentration of renal cytosolic aldosterone receptors (type I, aldosterone specific) had already increased at 6 wk of age in M-SHRSP, with no difference in affinity, and levels remained increased thereafter. There were no significant differences in molecular weights or ionic charges of either "activated" or "non-activated" aldosterone-receptor complexes between M-SHRSP and WKY, indicating that the molecular properties were similar in both groups. These results suggest that the increased concentration of aldosterone receptors in the kidneys of M-SHRSP might increase their aldosterone responsiveness and contribute to the development of high blood pressure in these animals.

Effects of mebendazole on Angiostrongylus costaricensis in mice, with special reference to the timing of treatment.

Mebendazole was given to mice infected with Angiostrongylus costaricensis at a single dose of 5 mg/kg at 6, 11, 16 or 21 days post-infection (p.i.) and in five successive doses at 5 mg/kg daily at 6, 11 or 16 days p.i. The effects were comparatively assessed by examining various parameters in host mice and worms. As a whole, the effects of mebendazole were caused more conspicuously by five successive treatments than by a single treatment. In both treatment modalities, the effects were more remarkable in earlier treatments, and nearly complete effects were caused by five successive treatments before 15 days p.i. These results suggest that the inhibition of egg formation and/or oviposition will inhibit the pathological changes caused in the disease by A. costaricensis, especially before the onset of the changes.

Sphingolipids of a cestode Metroliasthes coturnix.

Sphingolipids of Metroliasthes coturnix were studied. The cestode contained no detectable amounts of sphingomyelin. The major glycosphingolipids found were monogalactosylceramide, galactosyl-alpha-1-4-galactosylceramide, galactosyl-beta-1-6-galactosyl-beta-1-6-galactosylceramide and galactosyl-beta-1-6-galactosyl-beta-1-6-galactosyl-beta-1-6-galactosylce ramide. Their ceramides were mostly composed of C18-20 sphinganine or 4-D-hydroxysphinganine, which is N-acylated by extraordinarily long normal and 2-hydroxy acids such as C26 acid (range C16 to C30) except tetragalactosylceramide. The structure of glycosphingolipids was confirmed after their chromatographic separation into each molecular species using a novel analytical device, fast atom bombardment-mass spectrometry linked with reversed-phase high-performance liquid chromatography.

[Diaphragmatic fatigue under inspiratory resistive load in healthy subjects and the effect of several selections of frequency bands on the H/L (high/low) ratio].

In seven normal adult males the authors studied diaphragmatic fatigue by analysis of the power spectrum of the diaphragmatic electromyogram obtained with an esophageal electrode, and of the change of transdiaphragmatic pressure. Diaphragmatic fatigue was produced by inspiring through a resistive load, and mean value of the index of the tension time of the diaphragm (TTdi) maintained until fatigue was 0.33 +/- 0.09, which has been known to produce fatigue. The mean value of ratios of power at high frequencies to power at low frequencies (H/L) decreased to 61.2 +/- 16.8% of the initial value at just before breaking time. Breaking time was 4.88 +/- 1.37 minutes in average. The selection of low and high band widths for analysis of H/L ratio has differed slightly in the literature, but such differences do not have any substantial influence on the estimation of the changes of H/L ratio.

[A case of fatal asthma induced by timolol eye-drop].

A 74 year-old man, a known asthmatic since 1972, was treated by timolol eye-drop for acute glaucoma. Several hours later the patient developed a severe attack of asthma, and died subsequently. The autopsy findings were consistent with those of typical status asthmaticus. Our patient is, to our knowledge, the first case of fatal asthma induced by timolol eye-drop in Japan.