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Biomedical subjects

S Kuroki

Publications and source records attributed to S Kuroki.

At least 37 records · Page 2Linked to original sources

Calpastatin domain L is involved in the regulation of L-type Ca2+ channels in guinea pig cardiac myocytes.

We have found previously that L-type Ca2+ channel run-down in cell-free patches is partially (10-28%) reversed by calpastatin (CS) and have suggested that CS, an endogenous inhibitor of calpain, has a Ca2+-channel-regulating function. CS is composed of repetitive domains 1-4 (calpain-inhibitory domain) and domain L (a domain whose function is unknown). We therefore investigated which domain of CS was involved in the regulation of Ca2+ channel activity in guinea pig cardiac myocytes using the patch-clamp technique. After the patches were excised into inside-out mode in basic internal solution, the Ca2+ channel activity ran down to 0.45% of the control level recorded in the cell-attached mode. Application of human recombinant full-length CS (25 microM) and domain L (25 microM) restored the Ca2+ channel activity to 13 and 19% of the control level, respectively, while the channel activity was not restored by CS domain 1 (25 microM) (0.66%). Mouse CS domain XLL (25 microM), a complex of domain XL and domain L, restored the calcium channel activity to 11% of the control level. These results suggested that the Ca2+ channel-regulating function of CS is located in domain L. This study is the first description of the function of CS domain L.

Animals↗

A study of conformational stability of polyglycine and poly(L-alanine), and polyglycine/poly(L-alanine) blends in the solid state by (13)C cross-polarization/magic angle spinning NMR.

13C Cross-Polarization/Magic Angle Spinning nmr and T(1rhoH) experiments of polyglycine (PG), poly(L-alanine) (PLA), and PG/PLA blends prepared from dichloroacetic acid solution have been carried out, in order to elucidate the conformational stability of these polypeptides in the solid state. From these experimental results, it was clarified that the conformations of PG and PLA in their blends are strongly influenced by intermolecular hydrogen-bonding interactions that cause their miscibility at the molecular level.

Carbon Isotopes↗

Administration of bacterial lipopolysaccharide to rats induces heme oxygenase-1 and formation of antioxidant bilirubin in the intestinal mucosa.

Heme oxygenase (HO)-1, the rate-limiting enzyme in heme degradation, is induced by oxidative stress and its major end product, bilirubin, is a potent physiological antioxidant. We studied the induction of HO-1 and bilirubin production in intestinal mucosa using a rat model of sepsis. E. coli lipopolysaccharide was administered intraperitonealy to male Wistar rats and intestinal mucosa was harvested. Intestinal lipid peroxides increased significantly at 1 hr and peaked at 170% of the control value at 5 hr. GSH significantly decreased at 3 hr, reaching the nadir of 50% of the control value at 5 hr. HO-1 mRNA was maximally induced fivefold at 3 hr and HO-1 protein maximally increased to 10 times the control value at 7.5 hr. Both bilirubin and bilirubin oxidative metabolites were maximally increased at 10 hr, to 4.3 and 3.7 times the control value, respectively. These data suggest that oxidative stress in sepsis quickly induces HO-1 in intestinal mucosa and that subsequent production of bilirubin works as an antioxidant. The small intestinal mucosa is an active participant in the general response to sepsis.

Animals↗

Gadolinium pretreatment decreases survival and impairs liver regeneration after partial hepatectomy under ischemia/reperfusion in rats.

BACKGROUND: Modulation of Kupffer cell functions by treatment with gadolinium chloride protects the liver against reperfusion injury. However, its effect on liver regeneration after hepatectomy under ischemia/reperfusion has not been studied. Using a common clinical ischemia/reperfusion technique, we examined the effect of gadolinium on liver regeneration after hepatectomy in rats. METHODS: After an initial 15-minute ischemia and 15-minute reperfusion, 70% hepatectomy was performed during the second 15-minute ischemia period in gadolinium-pretreated (gadolinium group) and saline solution--pretreated (control group) rats. The 24-hour survival rate, relative liver weight, DNA synthesis rate, and hepatic adenosine triphosphate level were examined immediately after hepatectomy and on postoperative days (PODs) 1, 2, 3, and 7. Serum levels of total bilirubin, glutamic pyruvic transaminase, and endotoxin were also measured. RESULTS: The 24-hour survival rate was significantly lower in the gadolinium group (67%) than in the control group (100%). On POD 1, the relative liver weight and DNA synthesis rate were significantly lower in the gadolinium group than in the control group. On POD 1, serum total bilirubin and endotoxin levels were significantly higher in the gadolinium group than in the control group. Immediately after hepatectomy, the hepatic adenosine triphosphate level was significantly lower in the gadolinium group than in the control group. CONCLUSIONS: Under ischemia/reperfusion, gadolinium pretreatment impairs liver regeneration and energy status after hepatectomy and decreases postoperative survival.

Adenosine Triphosphate↗

Effects of cholestyramine on hepatic cholesterol 7alpha-hydroxylase and serum 7alpha-hydroxycholesterol in the hamster.

Cholestyramine increases activities of hepatic cholesterol 7alpha-hydroxylase and serum levels of 7alpha-hydroxycholesterol. To examine if serum 7alpha-hydroxycholesterol levels parallel with enzyme activity, 0, 0.5, 1, 2, 4, and 10% of cholestyramine was administered to female golden Syrian hamsters for 28 d in the dose-dependent study, and 2% cholestyramine for 0, 1, 3, 7, 14, 21, and 28 d in the time-dependent study. In the dose-dependent study, hepatic and serum cholesterol levels were significantly decreased dose-dependently when more than 0.5% of cholestyramine was fed for 28 d. Cholestyramine increased the cholesterol 7alpha-hydroxylase activity in a dose-dependent manner, while the serum 7alpha-hydroxycholesterol level was essentially unchanged. No correlation was found between the serum level and the hepatic enzyme activity. In the time-dependent study, hepatic and serum cholesterol levels markedly decreased when 2% cholestyramine was fed for longer than 3 d. The serum triglyceride level increased significantly for the first 7 d and then decreased. Cholesterol 7alpha-hydroxylase activity increased significantly as early as day 1, reached maximum activity level on day 7, and then kept the significantly high values until day 28. The serum 7alpha-hydroxycholesterol level significantly increased for the first 7 d and decreased to the pretreatment level thereafter. 7Alpha-hydroxycholesterol levels significantly correlated with serum cholesterol and triglyceride levels. We conclude that the serum 7alpha-hydroxycholesterol level does not always reflect the activity of hepatic cholesterol 7alpha-hydroxylase, when cholesterol metabolism is severely disturbed by cholestyramine.

Animals↗

Coronary artery disease and endothelial nitric oxide synthase and angiotensin-converting enzyme gene polymorphisms.

Nitric oxide (NO) inhibits platelet aggregation, proliferation of vascular smooth muscle cells, and leukocyte adhesion to endothelial cells. Thus, genetic variation in or near NO synthase may be involved in the pathogenesis of coronary artery disease (CAD). We investigated the association between CAD and endothelial constitutive NO synthase (ecNOS) and angiotensin-converting enzyme (ACE) gene polymorphisms by polymerase chain reaction analysis. These genotypes were examined in consecutive patients with CAD (n = 40) and control subjects (n = 34). The frequency of ecNOS genotypes in the CAD group (4a/a + 4a/b: 48%; 4bb: 52%) did not differ from that in the control group (4a/a + 4a/b: 29%; 4bb: 71%). The allele frequencies of ecNOS4a and ecNOS4b also did not differ between the two groups. On the other hand, the frequency of the ACE DD genotype in the CAD group (DD: 45%; ID: 35%; II: 20%) was significantly higher than that in the control group (DD: 21%; ID: 35%; II: 44%). The frequency of the D allele in patients with CAD (0.63) was also significantly higher than in controls (0.38). Combined analysis showed that the frequency of the ecNOS genotypes in ACE DD genotype subjects (4a/a + 4a/b: 39%; 4b/b: 61%) in the CAD group was not significantly different from that in the control group (4a/a + 4a/b: 29%; 4b/b: 71%). The ACE genotype might be a predictor of CAD, while the ecNOS genotype appeared to confer no appreciable increase in the risk of CAD.

Case-Control Studies↗

Prevalence and genetic diversity of cagD, cagE, and vacA in Helicobacter pylori strains isolated from Japanese patients.

BACKGROUND: Although cagD and cagE (cagDE) identified upstream of cagA have been shown to be involved in the induction of interleukin (IL)-8 expression, the relationship between cagDE status and gastroduodenal diseases still remains to be examined. Thus we investigated prevalence and genetic diversity of cagD, cagE, and vacA in Helicobacter pylori strains isolated from patients with peptic ulcer or gastritis. METHODS: We analyzed 73 H. pylori strains isolated from Japanese patients (gastritis (GA), 15; gastric ulcer (GU), 28; duodenal ulcer (DU), 23; GU and DU, 7). The presence of cagDE was evaluated by polymerase chain reaction (PCR) and Southern hybridization. The vacA genotype was examined by PCR, using type-specific primers. RESULTS: cagDE was present in 13 (86.7%) of 15 patients with GA, 26 (92.9%) of 28 patients with GU, 21 (91.3%) of 23 patients with DU, and 6 (85.7%) of 7 patients with GU and DU (P = 0.89). vacA signal sequence type s1 was found in 14 (93.3%) of 15 patients with GA, 26 (92.9%) of 28 patients with GU, 22 (95.7%) of 23 patients with DU, and 6 (85.7%) of 7 patients with GU and DU (P = 0.84). Sequences of cagDE and vacA in our Japanese strains were highly homologous with one another, and there were no disease-specific mutations. CONCLUSIONS: Most of the H. pylori strains in Japan were cagDE-positive, vacA s1 type, regardless of clinical outcome. The present study also indicated that these genes were conserved well among our H. pylori isolates.

Adult↗

Cloning and expression of the Ca2+ channel alpha1C and beta2a subunits from guinea pig heart.

Complimentary DNA clones encoding the alpha1C and beta2a subunits of guinea-pig cardiac L-type Ca2+ channels were isolated using the PCR method. The open reading frame encoded 2,169 amino acids for the alpha1C and 597 amino acids for the beta2a subunit. The proteins showed 94.2 and 94.8%, respectively, identity to the respective subunit of the rabbit protein. The message size of the guinea pig alpha1C and beta2a subunits was 8.0 and 3.5/4.0 kb, respectively. RT-PCR analysis revealed that the alpha1C subunit is expressed exclusively in the heart, while the beta2a subunit is expressed in the heart, cerebellum, whole brain, and stomach. The alpha1C and beta2a subunits are transiently expressed in BHK (baby hamster kidney) cells, and the channel currents were studied using the whole-cell patch clamp technique in medium containing 30 mM Ba2+. In cells expressing alpha1C alone, the Ba2+ current was activated at -30 mV and more positive potentials and peaked at about 10 mV. The co-expression of beta2a with alpha1C did not affect the voltage-dependence of the current, but increased the peak current and accelerated current decay. In cells transfected with guinea pig alpha1C and rabbit beta1+alpha2/delta, a Ba2+ current comparable to those in native myocytes was observed. The Ba2+ current can be blocked completely by nifedipine and is enhanced 3-fold by Bay K 8644. On the other hand, neither forskolin nor okadaic acid affects the Ba2+ current, suggesting that cAMP-mediated modulation is not easily reproduced in transfected cells, unlike that seen in native cardiac myocytes.

Amino Acid Sequence↗

Preoperative internal biliary drainage is superior to external biliary drainage in liver regeneration and function after hepatectomy in obstructive jaundiced rats.

OBJECTIVE: To examine the differences in regeneration rates and functions of the liver at the time of and after hepatectomy in obstructive jaundiced rats with preoperative external and internal biliary drainage. SUMMARY BACKGROUND DATA: The significance of biliary drainage before surgery is controversial in patients with obstructive jaundice. METHODS: After biliary obstruction for 7 days, rats were randomly divided into three groups: obstructive jaundice and hepatectomy (OJ-Hx), external biliary drainage and hepatectomy (ED-Hx), and internal biliary drainage and hepatectomy (ID-Hx). The OJ-Hx group underwent hepatectomy without biliary drainage; the other two groups underwent hepatectomy after biliary drainage for 7 days. At the time of hepatectomy, all rats were provided with internal biliary drainage. On days 0, 1, 2, 3, and 7 after hepatectomy, the DNA synthesis rate and the concentrations of adenine nucleotides and malondialdehyde in the liver were determined as markers of the hepatic regeneration rate, energy status, and lipoperoxide concentration, respectively. Portal endotoxin concentrations were measured and serum hyaluronic acid concentrations were determined as an indicator of hepatic endothelial function. RESULTS: The relative liver weight was significantly higher in the ID-Hx group than in the OJ-Hx group on days 1, 3, and 7 after hepatectomy and than in the ED-Hx group on days 1 and 2. The rate of hepatic DNA synthesis was significantly higher in the ID-Hx group than in the OJ-Hx and ED-Hx groups on day 1. The rate was similar in the ED-Hx and ID-Hx groups on day 2 but was significantly higher than in the OJ-Hx group. The hepatic malondialdehyde concentration was significantly higher on day 1 in the ED-Hx group than in the other two groups. It was lowest in the ID-Hx group throughout the study. Both biliary drainage procedures lowered the portal endotoxin concentration and serum hyaluronic acid concentration at the time of hepatectomy. The serum hyaluronic acid concentration was lowest in the ID Hx group. Hepatic adenine triphosphate concentrations and energy charge levels were similar among the three groups. CONCLUSION: Although both external and internal biliary drainage before hepatectomy improved serum liver function tests, portal endotoxin concentration, and serum hyaluronic acid concentration at the time of surgery, preoperative internal biliary drainage was superior to external drainage, as evidenced by the better liver regeneration and function after hepatectomy.

Animals↗

Effect of vaginal delivery on the Q-Tc interval in a patient with the long Q-T (Romano-Ward) syndrome.

Twelve ECG leads were monitored continuously during peripartum in a 23-year-old Japanese woman diagnosed as having the long Q-T (Romano-Ward) syndrome. Corrected Q-T (Q-Tc) intervals determined by 2 investigators blinded from the clinical informations disclosed that the Q-Tc interval increased during labor, suggesting that physical and/or emotional stress during labor might cause prolonged Q-Tc intervals in women with the long QT syndrome.

Adult↗

Bilirubin as an anti-oxidant for surgical stress: a preliminary report of bilirubin oxidative metabolites.

BACKGROUND: Bilirubin has been recognized as an anti-oxidant. The purpose of this study was to examine whether bilirubin would act as an antioxidant for surgical stress in humans. MATERIALS AND METHODS: Serum bilirubin and urinary bilirubin oxidative metabolites (BOM) were measured in 96 patients who underwent surgery. The antioxidant activity of bilirubin was assessed using BOM measured by enzyme-linked immunosorbent assay with an anti-bilirubin monoclonal antibody. RESULTS: Serum bilirubin levels increased after surgery in all 96 patients (p < 0.01), but did not correlate with operation time or blood loss (p = 0.53 and p = 0.28, respectively). BOM increased only in patients with major surgeries (p = 0.048). Significant correlations between BOM and operation time and blood loss were found (p < 0.01). CONCLUSIONS: Bilirubin appears to act as an antioxidant for invasive surgery in humans. Urinary BOM could be a reliable marker for the degree of surgical stress.

Adolescent↗

Broader clinical spectrum of Fukuyama-type congenital muscular dystrophy manifested by haplotype analysis.

Fukuyama-type congenital muscular dystrophy, Walker-Warburg syndrome, and muscle-eye-brain disease are clinically similar autosomal-recessive diseases, characterized by congenital muscular dystrophy, cobblestone lissencephaly, and eye anomalies. The classification of these disorders remains controversial. We analyzed five patients with congenital muscular dystrophy from four families who had severe eye and brain anomalies, such as retinal dysplasia and hydrocephalus, using polymorphic microsatellite markers flanking the Fukuyama-type congenital muscular dystrophy locus on chromosome 9q31. All patients were heterozygous for the Fukuyama muscular dystrophy founder haplotype with 3-kb insertion. In three cases, the other chromosome without the 3-kb insertion exhibited the same haplotype with a nonsense mutation on exon 3 of the Fukuyama gene. Thus, these three patients were compound heterozygotes for the condition. Severe eye anomalies such as retinal dysplasia or detachment and hydrocephalus could be included in the clinical spectrum of Fukuyama muscular dystrophy. The clinical spectrum of this disease is much broader than previously presumed.

Abnormalities, Multiple↗

Anti-GalNAc-GD1a antibody-associated Guillain-Barré syndrome with a predominantly distal weakness without cranial nerve impairment and sensory disturbance.

The serum antibodies to N-acetylgalactosaminyl GD1a (GalNAc-GD1a) and other gangliosides as well as to Campylobacter jejuni were determined in 147 patients with Guillain-Barré syndrome (GBS). We found a distinctive clinical pattern in patients with anti-GalNAc-GD1a antibodies compared with those without the antibodies, that is, lack of cranial nerve involvement (87% versus 38%), distal-dominant weakness (80% versus 25%), and no sensory disturbance (73% versus 22%). The frequency of distal-dominant weakness was significantly higher in patients with both C. jejuni infection and anti-GalNAc-GD1a positivity (100%) than in C. jejuni-negative/anti-GalNAc-GD1a-positive (25%), C. jejuni-positive/anti-GalNAc-GD1a-negative (32%) and C. jejuni-negative/anti-GalNAc-GD1a-negative patients (20%). Lack of cranial nerve involvement and sensory disturbance were found in most C. jejuni-positive/anti-GalNAc-GD1a-positive and C. jejuni-negative/anti-GalNAc-GD1a-positive patients, but not in C. jejuni-positive/anti-GalNAc-GD1a-negative and C. jejuni-negative/anti-GalNAc-GD1a-negative patients. Although the anti-GM1-positive/anti-GalNAc-GD1a-negative patients mostly (75%) lacked cranial nerve involvement, distal-dominant weakness (38%) and lack of sensory disturbance (13%) were infrequent. These results may indicate that (1) the combination of C. jejuni infection and anti-GalNAc-GD1a antibodies, but not anti-GalNAc-GD1a, anti-GM1, or C. jejuni infection alone, is associated with a predominantly distal weakness, (2) the presence of anti-GalNAc-GD1a, rather than C. jejuni infection or anti-GM1 antibody, is associated with a lack of sensory disturbance, (3) both anti-GalNAc-GD1a and anti-GM1 antibodies are independently associated with a lack of cranial nerve impairment.

Acetylgalactosamine↗

[A case of synchronous bilateral double primary lung cancer successfully treated by Nd-YAG laser therapy and chemotherapy].

A 61-year-old man was admitted to our hospital with the chief complaint of hemosputum. He was a heavy smoker. A chest radiograph revealed a tumor shadow in right S3 which invaded the pulmonary artery. There was also an associated patchy shadow in the periphery as well as ipsilateral pleural effusion. Bronchoscopy revealed a near occlusion with superficial infiltration at the right B3 bronchus and a nodular tumor at the bifurcation between the left upper and lingual division bronchi, which was consistent with endoscopic early lung cancer. Although both tumors were histologically diagnosed as squamous cell carcinoma, this was considered to be a case of synchronous double primary lung cancer due to their mutual isolation. The left tumor was subsequently diagnosed as carcinoma in situ. Following Nd-YAG laser therapy for carcinoma in situ and 4 courses of systemic chemotherapy using TXT and CDDP, bronchoscopy revealed no residual cancerous tissue and no tumor apart from the nodular shadow in right S3 was seen on a radiograph. Subsequent right upper and middle lobectomy and lymph node (R2a) dissection revealed no residual tumors.

Antineoplastic Combined Chemotherapy Protocols↗

Run-down of L-type Ca2+ channels occurs on the alpha 1 subunit.

Run-down of L-type Ca2+ channels in CHO cells stably expressing alpha 1c, alpha 1c beta 1a, or alpha 1c beta 1a alpha 2 delta gamma subunits was studied using the patch-clamp technique (single channel recording). The channel activity (NPo) of alpha 1c channels was increased 4- and 8-fold by coexpression with beta 1a and beta 1a alpha 2 delta gamma, respectively. When membranes containing channels composed of different subunits were excised into basic internal solution, the channel activity exhibited run-down, the time-course of which was independent of the subunit composition. The run-down was restored by the application of calpastatin (or calpastatin contained in cytoplasmic P-fraction) + H-fraction (a high molecular mass fraction of bovine cardiac cytoplasm) + 3 mM ATP, which has been shown to reverse the run-down in native Ca2+ channels in the guinea-pig heart. The restoration level was 64.7, 63.5, and 66.4% for channels composed of alpha 1c, alpha 1c beta 1a, and alpha 1c beta 1a alpha 2 delta gamma, respectively, and was thus also independent of the subunit composition. We conclude that run-down of L-type Ca2+ channels occurs via the alpha 1 subunit and that the cytoplasmic factors maintaining Ca2+ channel activity act on the alpha 1 subunit.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Genetic contribution of the tumor necrosis factor region in Guillain-Barré syndrome.

We studied genetic polymorphisms in the tumor necrosis factor (TNF) region in 81 Japanese patients with Guillain-Barré syndrome (GBS) and 85 controls. A significantly higher frequency of the 100-base pair (bp) (TNFa2) allele of the TNFa microsatellite marker, which is associated with high TNF alpha production, existed in Campylobacter jejuni-positive (Cj+) GBS patients than in controls, suggesting the involvement of a genetic predisposition to high TNF alpha secretion in the development of C. jejuni-related GBS.

Alleles↗