[Ventricular septal defect, pulmonary hypertension and 3C (cranio-cerebello-cardiac) syndrome: report of a case].

We report a case with the rare association of craniofacial anomalies, cerebellar vermis hypoplasia (Dandy-Walker variant) and congenital heart disease referred to as 3C (cranio-cerebello-cardiac) syndrome. A male infant was born at 40 weeks' gestation. The birth weight was 2,896 g. At birth he had macrocephaly, widely open metopic suture, ocular hypertelorism, cleft palate, apparently low-set ears, cerebellar vermis hypoplasia and ventricular septal defect (VSD) with pulmonary hypertension. Chromosomes were normal and we diagnosed 3C syndrome. At 8 months of age we performed a cardiac catheterization. The pulmonary artery pressure was 70/26 (48) mmHg and pulmonary vascular resistance (RpU) was 8.5 U/m2. But under the oxygenation RpU decreased to 1.5 U/m2. At 10 months of age we performed VSD patch closure. After operation his pulmonary artery pressure was 39/13 (25) mmHg under the oxygen therapy. He was on good course and discharged at 58 post operative day.

Recurrence of Progressive Familial Intrahepatic Cholestasis Type 2 Phenotype After Living-donor Liver Transplantation: A Case Report.

BACKGROUND: Progressive familial intrahepatic cholestasis 2 (PFIC2) is the result of mutations in the ABCB11, which encodes for bile salt export pump (BSEP). An absence of BSEP in the canalicular membrane causes cholestasis and leads to the development of end-stage liver disease in the first decade of life. Liver transplantation (LT) has been considered curative for BSEP disease. However, patients with PFIC2 having undergone LT have recently been reported to develop recurrence of cholestasis together with the clinical and histological features of primary BSEP disease. CASE REPORT: We herein present a rare case of a patient with PFIC2 who developed post-transplantation recurrence of progressive intrahepatic cholestasis due to antibodies against BSEP after living-donor LT, which mimicked primary BSEP disease. The patient had mutations in the ABCB11 gene, resulting in the complete absence of BSEP in the native liver, explaining the lack of tolerance. Immunofluorescence staining of normal human liver sections with the patient's serum and using an anti-human immunoglobulin G antibody to detect serum antibodies showed reactivity to the BSEP epitope in the canalicular membrane. We suggest that the patients having undergone LT had been associated with a risk of autoantibody formation against the BSEP protein. The absence of primary tolerance for the BSEP epitopes may explain the formation of the anti-BSEP antibodies after LDLT.

Formation of inositol phosphates mediated by M3 muscarinic receptors in type-1 and type-2 astrocytes from neonatal rat cerebral cortex.

Muscarinic receptor subtype in type-1 and type-2 astrocytes from rat neonalal cerebral cortex was examined for carbachol-elicited inositol phosphate (IP) formation. The formation of carbachol-elicited IP was inhibited by various muscarinic antagonists in the following relative order of potency: 4-DAMP > or = atropine >> pirenzepine > AF-DX 116. This pharmacological profile suggests that the activation of the M3 muscarinic receptor subtype is responsible for the stimulation of IP formation in both astrocytes.

[Incidence of hip fracture and prognosis in Ehime Prefecture].

Hip fracture among the elderly has increasingly attracted public health concern in Japan. For the purpose of revealing the epidemiological features of hip fracture, post-treatment prognosis, and related factors, we studied all cases of hip fracture which occurred in Ehime Prefecture during 1992 who were admitted to hospitals or clinics, and followed up the cases for a subsequent two years. The incidence rate of hip fracture was 29.2 per 100,000 for males, and 84.0 for females, which are much lower than in Europe and the Untied States. Compared with rates reported previously in Japan, the age--specific incidence rates for males in Ehime were almost identical to the respective rates estimated by a nationwide survey and the rates for Tottori. However, the rate for males 80 years old or more was found to be lower than the corresponding rate for Kagawa. In females, the age--specific incidence rates for Ehime were similar to the Japanese averages, and the rates for Tottori and Kagawa. Falling from a standing position was a leading cause of hip fracture among older patients. Osteosynthesis was the main treatment modality elected trochanteric fractures. On the other hand, prosthetic replacement was predominant in cervical fracture. Multivariate analysis using a multiple logistic model showed that medical facility category, age, type of treatment, and cause of fracture had statistically significant relations to mobility at the time of discharge. Of patients who could walk at the time of discharge, 83.7% (159/190) were alive two years after discharge. Multiple logistic model analysis identified gender as a significant contributing factor to this mortality. Among the survivors, 81.8% (130/159) retain the ability to walk. Logistic model analysis revealed that older age experience a significantly higher risk in losing mobility.

Isolation of guinea pig gastric smooth muscle cells.

A method of isolating guinea pig gastric smooth muscle cells was established in order to investigate gastric contractile mechanisms at the cellular level. Strips of gastric muscle were digested enzymatically with collagenase and dispase, and the smooth muscle cells isolated on a percoll density gradient, with a purity of 80-90%. The muscle cells were over 90% viable and demonstrated a remarkable contractile response to carbachol stimulation in a dose-dependent manner, from 10(-7) M to 10(-4) M. Intracellular Ca2+ concentration following carbachol stimulation was monitored, since Ca2+ is essential to the contractile process, and it was found to increase almost in parallel with contraction. These results suggest that the measurement of intracellular Ca2+ concentration is useful for analyzing the contractile mechanism of isolated smooth muscle cells.

[Sensitive detection of K-ras oncogene codon 12 mutations by nested PCR using mismatched primers and selective digestion of non-mutated PCR fragments with restriction enzyme].

Sensitive detection of K-ras oncogene mutation at codon 12 using nested polymerase chain reactions (PCR) with mismatched primers combined with selective digestion of nonmutated DNA fragments after the first PCR amplification was reported by Levi S, et al. (Cancer Res 51:3497-3502, 1991). We examined the usefulness of this novel technique in cell lines carrying various K-ras codon 12 mutations and clinical samples such as colon carcinoma tissue, corresponding normal colonic mucosa and pancreatic juice obtained from the patients with pancreatic carcinoma undergoing endoscopic retrograde pancreatography (ERP). In analysis of tumor cell lines carrying mutated K-ras at codon 12, the fragment length of the amplified DNA was 135 bp after digestion using restriction enzyme BstN1, whereas those of nonmutated cell lines were 106 bp. This method was highly sensitive to detect mutant DNA diluted at a ratio of 2048 fold with normal DNA samples obtained from peripheral blood lymphocytes. In analysis of clinical samples, 4 out of 10 colorectal carcinoma tissues were positive for K-ras gene mutation at codon 12, however, no mutations were detected in corresponding normal colonic mucosa. In analysis of pancreatic juice taken from the patients with pancreatic carcinoma, 1 out of 3 samples were positive for K-ras mutation at codon 12. Thus, this novel approach was thought to be useful for detection of minimum number of cancer cells from compound samples containing larger amounts of normal cells.

[Comparative studies and application of a simple method of extraction of RNA from hepatitis C virus].

Conventional RNA extraction methods, such as SDS/phenol and AGPC methods involve time consuming and complicated manipulation procedures, which could be a cause of contamination. In this study, we compared the conventional methods with a method using a newly developed reagent, "SepaGene-RV" to extract RNA. The results obtained by the new method were the same as those obtained by the conventional methods in the comparative experiments with the sera of patients infected with HCV and detection limit tests using recombinant HCV genome. The sensitivity to detect HCV-RNA with SepaGene-RV was equivalent to that of conventionally used methods to extract RNA, and the manipulation procedure was simpler and it was less time consuming. In addition, we could detect plus-stranded and minus-stranded HCV-RNA by polymerase chain reaction assay (PCR) from liver tissue obtained from liver biopsy and peripheral blood mononuclear cells (PBMC). We confirmed that HCV infects not only liver tissue but also PBMC where replication occurs.

[A case of idiopathic pulmonary hemosiderosis of adult onset].

A 51-year-old man presenting with hemoptysis was admitted to our hospital. Chest radiography revealed air space consolidation in the right lung field. Laboratory data showed anemia, hypoxemia, and no evidence of inflammatory signs, bleeding tendency, renal dysfunction, or collagen vascular diseases. Tests of anti-GBM antibody, P-ANCA, and C-ANCA were negative. Microscopic examination of the lung tissue specimens obtained by video assisted thoracic surgery revealed hemorrhage and numerous hemosiderin-laden macrophages in the alveoli. No deposition of immunoglobulin and vasculitis were seen. These findings were consistent with a diagnosis of idiopathic pulmonary hemosiderosis. Steroid therapy had a limited effect, and the patient died. Idiopathic pulmonary hemosiderosis of adult onset is rare in Japan.

[Myasthenia gravis presenting as isolated respiratory failure].

A 61-year-old woman in excellent health and taking no medications presented with progressive dyspnea on exertion. She did not exhibit diplopia, dysphagia, dysarthria or muscle weakness. Her condition deteriorated, and respiratory distress developed. The patient was intubated and placed on mechanical ventilation. A computed tomogram of her chest revealed a moderately enlarged thymus gland unsuspected on a plain x-ray film. Myasthenia gravis was diagnosed on the basis of a high acetylcholine receptor antibody titer. Treatment started with prednisolone, anticholinesterase agent, and plasma exchange. The patient underwent a thymectomy 3 weeks after her diagnosis. Her symptoms were brought under control by anticholinesterase agent and prednisolone. This case illustrates the need to consider myasthenia gravis as well as other motor-neuron disorders when evaluating individuals presenting acute respiratory failure of unknown origin.

Conversion to prolonged-release tacrolimus for pediatric living related donor liver transplant recipients.

Prolonged-release tacrolimus allows for once-daily dosing. Although many adult recipients have been switched from standard tacrolimus, prolonged-release tacrolimus has not been popular for pediatric patients despite the potential benefits for medication compliance. We report on prolonged-release tacrolimus for 11 pediatric living related donor liver transplant (LRDLT) recipients. Patients under 18 years of age who were receiving standard tacrolimus-based immunosuppression and steroid taper underwent conversion from standard to prolonged-release tacrolimus. We monitored tacrolimus trough levels and liver function tests (LFTs). We also assessed adverse effects and satisfaction levels for prolonged-release tacrolimus. Mean age at transplantation was 4.3 years. The mean duration of follow-up was 12 months. The ratios of trough levels with prolonged-release vs standard tacrolimus were 0.97, 0.95, and 0.92 at 1, 2, and 4 weeks post conversion, respectively. Two patients discontinued prolonged-release tacrolimus owing to abnormal LFTs and neurological abnormalities, respectively; but symptoms resolved after reconversion. One patient returned to standard tacrolimus and the other was converted to cyclosporine. Once-daily administration satisfied 89% of patients. In the overall assessment, conversion to prolonged-release tacrolimus satisfied all patients. Prolonged-release tacrolimus was useful for pediatric patients after LRDLT. Trough levels after conversion were compatible with those before conversion. Most patients were satisfied with prolonged-release tacrolimus. However, some patients failed conversion because of unexpected responses. Close observation after conversion is required even if patients have previously had an uneventful course on standard tacrolimus.

Healthy lifestyles are associated with higher natural killer cell activity.

BACKGROUND: To investigate the association of individual lifestyle with immune response against tumors, we assayed peripheral blood lymphocytes of 62 healthy males ranging in age from 30 to 60 years for natural killer (NK) cell activity, which is the first line of defense against tumors, and for frequencies of NK cell subsets. METHODS: The NK cell activity was determined by 51Cr release assay, and NK cell subsets were counted using surface antigens (CD16, CD57) which recognize NK cell subgroups with different cytolytic potentials. Subjects were classified into groups reporting good, moderate, and poor lifestyles according to their responses on a questionnaire regarding eight health practices (tobacco smoking, alcohol consumption, hours of sleep, physical exercise, eating breakfast, balanced nutrition, hours of work habits, and mental stress). RESULTS: Individuals reporting good lifestyle habits were found to have the highest NK cell activity and it was significantly higher than the NK cell activity in those reporting poor lifestyle habits. Those reporting good health practices regarding smoking and physical exercise showed significantly higher NK cell activity at an effector-to-target-cell ratio of 40:1. CONCLUSIONS: The higher NK cell activity among individuals reporting good lifestyle practices may reflect an increase in the cytolytic potential of NK cell activity within the CD16+, CD57- subset.

Health status, life habits, and social background among the JPHC study participants at baseline survey. Japan Public Health Center-based Prospective Study on Cancer and Cardiovascular Diseases.

A self-reported questionnaire on the health status, life habits, and social background was conducted at baseline in the Japan Public Health Center-based Prospective Study on Cancer and Cardiovascular Diseases (JPHC study). This report presents the outcome of the study regarding past or family history of various diseases, medical treatment, life habits such as physical labor or sports, and social background among study participants. In both cohorts I and II, prevalent past and family history included hypertension, stroke, and cancer, whereas the prevalence of coronary heart diseases was historically low. The prevalence of a past history of hypertension and stroke was higher in the northern part of Japan, Ninohe, and Yokote, and lower in Okinawa, compared to the other districts. The prevalence of participants with a history of stomach cancer and liver cancer was higher in Arikawa than in other districts. The frequency of participants who took medication from doctors ranged from 20% to 30%, higher in the Tohoku areas, and lower in Okinawa compared to the other districts. All districts showed a high rate of over 70% for the participation rate for basic health examination conducted by the local government, The rate was particularly high in the Tohoku area where a high prevalence of a history of hypertension was found. The frequency of persons who had a chance to participate in sports or physical exercise was high in Okinawa and Suita subcohort 2, although the mean total physical activity (both at work and for leisure time) was lowest in the latter subcohort. No substantial differences were found in compositions of personality among districts. The frequency of more active and positive persons, however, was relatively higher in urban areas and lower in Okinawa compared to the other districts. The association between the differences of health status, life habits, and social background and the occurrence of various lifestyle-related diseases will be clarified in a follow-up study within the JPHC study.

Histamine-induced inositol phosphate accumulation in type-2 astrocytes.

Histamine elicited dose-dependent accumulation of [3H]inositol phosphates in type-2 astrocytes, but not in type-1 astrocytes. The ED50 was about 2.4 x 10(-6) M and the maximal response was obtained at 10(-4) M. This response was dose-dependently inhibited by H1-antagonists, mepyramine and D- and L-chlorpheniramine. Furthermore, D- and L-chlorpheniramine showed stereoselectivity in the inhibition. On the other hand, an H2-antagonist, famotidine, and an H3-antagonist, thioperamide, did not inhibit the response. These results indicate that histamine stimulates accumulation of inositol phosphates in type-2 astrocytes via H1-receptors.

Endoscopic diagnosis of reflux esophagitis by the dye-spraying method.

Twenty-one patients (13 patients after gastric surgery and 8 patients with hiatus hernia or other esophageal disease) in whom reflux esophagitis had been diagnosed by standard endoscopy, and who had undergone Lugol-spraying endoscopy, were reviewed. Ninety-two biopsy specimens were obtained from the esophageal mucosa under direct vision and stained with hematoxylineosin and PAS. The coincidence rate of the findings of dye-spraying endoscopy with histological findings was 89.9% (80/89), as opposed to 69.7% (62/89) attained by standard endoscopy (p less than 0.001), indicating that Lugol-spraying endoscopy is highly compatible with histology. Of 34 biopsy specimens showing papillary hyperplasia, 18 (52.9%) were stained unevenly and lightly, and 14 (41.2%) evenly and lightly. Of nine biopsy specimens showing epithelial regeneration, eight (88.9%) were unstained. All 26 biopsy specimens with erosion/ulcer were also unstained. It is concluded that Lugol-spraying endoscopy is of value in diagnosing reflux esophagitis, especially in determining the intensity and extent of the disease.

[Investigation of etching and bonding on the inner surface of free enamel. SEM observation and thermal cycling test for pigment invasion].

The purpose of this study is to examine adhesive condition on the inner surface of free enamel. If it was possible to use an adhesive composite resin to reinforce free enamel, it would be unnecessary to remove the free enamel entirely. To this study, we have used a visible light-cured and adhesive composite resin for the inner surface of free enamel. At the first, we carried out SEM examination of adhesive condition on the inner surface of free enamel. Then we performed thermal cycling test for pigment invasion. Results 1. SEM observation found tag-like extensions on the inner surface of free enamel. These extensions were of 2 shapes: horseshoe and beehive. 2. Thermal cycling test for pigment invasion showed practically no difference in marginal leakage between the experimental free-enamel cavities and control box cavities. 3. As a result of observation of adhesive conditions on the inner surface of free enamel, we believe that a certain degree of reinforcement of that enamel can be expected from composite resin.

Two cases of hypertrophic cardiomyopathy with coronary vasospasm.

Chest pain in patients with hypertrophic cardiomyopathy seems to be caused by relative myocardial ischemia due to the left ventricular outflow pressure gradient and myocardial hypertrophy. However, in 2 cases of hypertrophic cardiomyopathy chest pain was associated with coronary vasospasm. Thus, chest pain in these cases was decreased not by a beta-blocker but by isosorbide dinitrate and a calcium antagonist. Because beta-blockers are commonly used for hypertrophic obstructive cardiomyopathy and chest pain may be aggravated by beta-blockers in patients with coronary vasospasm, a combination of beta-blocker, isosorbide dinitrate and calcium antagonist was necessary for this hypertrophic cardiomyopathy with variant angina.

Decreased left ventricular filling pressure 8 months after corrective surgery in a 55-year-old man with tetralogy of Fallot: adaptation for increased preload.

A 55-year-old man with tetralogy of Fallot underwent corrective surgery. Left ventricular filling pressure increased markedly with increased left ventricular volume one month after surgery, then decreased over the next 7 months, presumably due to increased left ventricular compliance.