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To study the expression of the amiloride-sensitive sodium channel, a putative mechano-receptor in the rat eye, reverse transcriptase-polymerase chain reaction and in situ hybridization were done. The gene for the alpha subunit of the amiloride-sensitive sodium channel was shown by polymerase chain reaction to be expressed in mRNA isolated from the whole eye tissue. In situ hybridization demonstrated that the gene was expressed in basal layers of the corneal and conjunctival epithelium, ciliary epithelial cells, lens epithelial cells at the equator, retinal and iris pigment epithelial cells, ganglion cells and cells in the inner and outer nuclear layers of the retina. The results suggest that the amiloride-sensitive sodium channel plays a role in maintaining sodium balance as well as in possible mechanosensation in these ocular tissues.

Nursing practitioners' perception of inpatients' anxiety, self-esteem, purpose-in-life and health locus of control.

X-ray computed tomography (CT) has been used for diagnosis of pulmonary emphysema because it can reveal the morphology of low attenuation areas. Recently, 99mTc-Technegas imaging, one of several types of scintigraphic techniques, has been used for ventilation scintigraphy. Technegas scintigraphy was performed on 15 patients with pulmonary emphysema, and we compared the extent and degree of abnormal findings on Technegas scintigraphy with the extent of low attenuation areas shown by CT. We classified the findings of Technegas imaging into three grades, from mild to severe, according to the extent of peripheral irregularity and central hot spot formation. We also classified the findings of CT as centrilobular emphysema into three grades from mild to severe according to the extent of low attention areas in the peripheral lung fields. In 5 cases, CT and Technegas assessment resulted in equivalent diagnoses. In eight cases, Technegas images showed more detailed findings than CT images. In the two remaining cases, which were diagnosed as panlobular emphysema on CT, Technegas images showed the severe stage. Technegas scintigraphy was useful for diagnostic assessment of pulmonary emphysema, especially for panlobular emphysema, which is difficult to distinguish from the normal lung condition by CT assessment.

Five patients with malignant pleural mesothelioma (MPM) were studied to determine whether CYFRA 21-1 is useful for diagnosis of this disease. In pleural effusions, the median concentration of CYFRA 21-1 from 4 patients with MPM was significantly higher than for 34 patients with benign diseases. The sensitivity of serum CYFRA 21-1 for diagnosis of MPM was 40% and its concentration changed in proportion to disease activity in all cases. Immunohistochemically, anticytokeratin 19 antibody revealed strong staining in both epithelial and sarcomatous MPM tissues. Based on these results, we conclude that measurement of CYFRA 21-1 in pleural effusions and serum may be useful for diagnosing and monitoring MPM.

Sulfate and taurine are the main metabolites of L-cysteine in mammals and are excreted in the urine. The effect of a high protein diet on the ratio of sulfate to taurine excretion was studied in rats using synthetic 25% (standard protein diet group, group A) and 40% (high protein diet group, group B) casein diets. Average taurine and sulfate excretions (mumol/kg of body weight per day) were 280.4 +/- 93.8 and 943.2 +/- 144.8 in group A and 553.4 +/- 124.5 and 2675.0 +/- 390.9 in group B, respectively. Thus, the average taurine/sulfate ratio in group A was 0.30 +/- 0.08. By a single administration of 5 mmol of L-cysteine/kg of body weight in group A, the average taurine and sulfate excretions increased to 1127.5 +/- 120.2 and 4043.0 +/- 305.6, respectively, but the taurine/sulfate ratio changed only slightly (0.28). The average taurine/sulfate ratio in group B was 0.22 +/- 0.07, a significantly lower ratio than that in group A, which means that daily intake of a high protein diet resulted in more sulfate excretion. The taurine/sulfate ratio in group B was affected only slightly (0.19) by the cysteine administration as well. These results suggest that the ratio of taurine and sulfate production was determined by dietary protein content and that the increase in sulfate production is larger than that of taurine production when the intake of dietary protein is increased.

Acute aortic dissection is a life-threatening condition, and may be treated with aggressive hypotensive drug therapy, but emergency surgery is often necessary. We evaluated the effectiveness of stent-grafts for the treatment of acute aortic dissection. Aortic dissection was surgically created in the descending thoracic aorta in 20 adult mongrel dogs. A stent-graft was inserted in the entry position. The tested animals were divided into 4 groups based on re-entry type and blood pressure alteration rate (AR) after acute aortic dissection. After insertion of the stent, the following results were observed: a) AR improved; b) proximal descending aorta and superior mesenteric arterial flows increased; c) cardiac function improved; and d) the dissecting aortic diameter decreased in the presence of pressure gradient group. From these results, insertion of a stent-graft to treat acute aortic dissection was judged to be effective.

Intrathymic (i.t.) injection of allogenic cells without administration of anti-lymphocyte serum (ALS) in neonatal recipients has induced donor-specific tolerance to subsequent cardiac allografts in rats. This study examines whether similar tactics can be successfully applied to a hamster-to-rat cardiac xenotransplantation model. Lewis neonates on their first day of life underwent i.t., subcutaneous (s.c.), intraperitoneal (i.p.), or intravenous (i.v.) injections of 5 x 10(7) Golden Syrian hamster splenocytes. After six weeks, the rats underwent heterotopic cardiac transplantation of hamster hearts. Cyclophosphamide (CyP) was administered on the day before surgery and postoperatively to suppress antibody-mediated graft rejection. Rats given splenocytes with 80 mg/kg of CyP had the following graft survival times: 8 to 12 days for i.t. injection (mean, 9.4 days); 5 to 7 days for s.c. injection (mean, 6.6 days); 4 to 11 days for i.p. injection (mean, 7.4 days); and 4 to 13 days for i.v. injection (mean, 7.9 days). Only the extension of graft survival produced by i.t. injection was statistically significant in comparison with the rats given only CyP treatment (mean, 7.5 days; P < 0.05). Thus, it appears that i.t. injection of xenogenic splenocytes in neonatal recipients with administration of CyP, but without ALS, can prolong xenograft survival. This biological intervention may be most useful in pediatric xenotransplantation when combined with other immunomodulation techniques.

The effects of a low protein diet on the excretion of sulfate and taurine, major metabolites of L-cysteine in mammals, were studied in rats fed with synthetic 10% (group A) and 25% (group B) casein diets. The average excretions of total taurine (taurine plus hypotaurine) and total sulfate (free plus ester sulfate) (mumol/kg of body weight per day after the adaptation to the synthetic diet) in group A were 14.2 +/- 13.4 and 122.3 +/- 39.6, respectively, which were very low compared with 280.4 +/- 93.8 and 943.2 +/- 144.8, respectively, in group B. The taurine/sulfate ratio in group A was 0.12 +/- 0.11, which was significantly lower than that (0.30 +/- 0.08) in group B. A single intraperitoneal injection of 5 mmol of L-cysteine per kg of body weight in group A resulted in an increase in average taurine and sulfate excretion to 693.4 +/- 195.6 and 2440.6 +/- 270.0, respectively, and thus the average taurine/sulfate ratio increased to 0.29. These increases were transient and low taurine excretion resumed again 24 h after the L-cysteine administration. L-Cysteine injection in group B resulted in a similar increase in taurine and sulfate excretion, but the ratio changed only slightly (0.28). The present results suggest that in vivo production of taurine is reduced preferentially over sulfate production when sulfur amino acid supply is limited.

To evaluate viral interference between hepatitis B and C, we studied coinfected patients serologically and molecular biologically. Twenty-seven patients positive for hepatitis B surface antigen (HBsAg) and anti-hepatitis C virus (HCV) antibody, were classified into Groups BC-L and BC-H according to DNA-polymerase activity (less or greater than 100 cpm, respectively). Patients with hepatitis B or C alone were also enrolled as controls. HCV-RNA was detected more often in Group BC-L than in Group BC-H. Genotype 1b of HCV was determined in 75% of Group BC-H, 87.5% of Group BC-L, and 70.7% of hepatitis C-only patients. Activity of DNA-polymerase in coinfected patients was lower in patients positive for HCV-RNA as compared with those negative. HBsAg titers tended to be lower in coinfected patients than in patients with hepatitis B virus (HBV) alone. In conclusion, in coinfection, HBV may suppress the replication of HCV and HCV appears to reduce the expression of HBsAg and probably suppresses HBV replication.</P>

In order to elucidate factors influencing the prognosis of small-cell lung cancer (SCLC), we reviewed the records of 253 patients with SCLC and evaluated 20 pretreatment prognostic factors by univariate analysis and Cox's multiple regression analysis. Recursive partitioning and amalgamation (RPA) was employed to identify subgroups with similar survival rates. Cox's multiple regression analysis identified five significant factors: extent of disease, number of metastatic sites, serum albumin, serum lactate dehydrogenase, and presence of weight loss. Among these, extent of disease was the most influential factor. RPA analysis revealed three subgroups predicting significantly different prognoses. The median survival time and 3-year survival rate were 18.4 months and 20.6%, respectively for the good-risk group (limited disease without weight loss), 13.5 months and 9.1%, respectively for the intermediate-risk group (limited disease with weight loss or extensive disease with less than two metastatic sites), and 9.2 months and 0%, respectively for the poor-risk group (extensive disease with two or more metastatic sites). These results will be useful for development of new staging system or subsequent stratification for randomized trials.

The carbohydrate-binding specificities of lectins in solution to glycoproteins and neoglycolipids immobilized on a solid phase were analyzed in order to establish a simple, rapid method for structural analysis of the carbohydrate moieties of small amounts of individual glycoproteins blotted on membrane. Eight glycoproteins containing typical O-linked tetrasaccharides or a series of typical N-linked oligosaccharides of the high-man-nose type, hybrid type, and complex type and 6 neoglycoproteins containing mono- or di-saccharides were dot blotted on membranes and the membranes were then reacted with 8 kinds of horseradish peroxidase-conjugated lectins before and after heat treatment. Neoglycolipids containing the glycoprotein-derived oligosaccharides immobilized on a thin layer chromatography plate were also reacted with lectins. The heat treatment of the membrane increased lectin reactivity toward the glycoproteins. The carbohydrate-binding behavior of lectins, Phaseolus vulgaris erythroagglutinin, wheat germ agglutinin, and concanavalin A in solution toward glycoproteins and neoglycolipids immobilized on a solid phase differed from that of immobilized lectins toward oligosaccharides in solution. This difference should be noted in lectin detection of specific carbohydrates of individual glycoproteins on membrane.

In this study we examined the effects of continuous calcium channel blocker (CCB) infusion on pancreatic duct-ligated acute pancreatitis (AP) in rabbits. Thirty rabbits were used for this study. Animals in group 1 (n = 10), which served as a control group, underwent dummy operations and received 0.5 microliter/h normal saline via the internal jugular vein. Animals in group 2 (n = 10) with artificially-induced pancreatitis received the same dosage of saline in the same manner. Animals in group 3 (n = 10) with artificially-induced pancreatitis received 180 micrograms/kg/h CCB (Verapamil) via the jugular vein starting from just before pancreatic duct ligation. AP histology score, plasma amylase levels and liver function tests were measured after 48 h. Verapamil infusion did not prevent the rise in plasma amylase levels, nor did it prevent pancreatic inflammation and damage. Serum levels of serum glutamate pyruvate transaminase, serum glutamate oxalacetate transaminase and alkaline phosphatase were significantly elevated in group 2 and significant reductions were seen in the Verapamil treated animals (group 3). The findings in this study imply that a continuous 180 micrograms/kg/h dose Verapamil infusion does not ameliorate the pathogenesis of pancreatitis induced by ligation of pancreatic duct but do not rule out a dose-dependent protective effect. Meanwhile, the lowering of liver function test scores should be considered the beneficial effect of CCBs, and this should be investigated in further studies.

Hepatocyte growth factor (HGF) and c-met proto-oncogene product (c-Met) have varied biological functions in different tissues and have been implicated in mitogenic, motogenic and morphogenic responses in both organ regeneration and carcinogenesis. Some studies have suggested that the overexpression of c-Met and epidermal growth factor receptor (EGFR) are associated with growth advantage, while transforming growth factor-beta receptor II (TGF beta R II) is associated with growth disadvantage of human prostatic adenocarcinoma. However, it is unclear if the expression of c-Met correlates with the expression of EGFR and TGF beta R II, and with the proliferative status of human prostatic adenocarcinoma. Using immunohistochemical staining with anti-c-Met (C-12), anti-EGFR (NCL-EGFR) and anti-TGF beta R II (L-21) antibodies, we determined the frequency of expression of c-MET, EGFR, and TGF beta R II respectively in a series of 134 radical prostatectomy specimens. We evaluated the relationship between the expression of these receptors and clinicopathological characteristics. Overall, c-Met immunostaining was detected in 54 of 134 (40.3%) cases, EGFR in 45 (33.6%) and TGF beta R II in 64 (48.4%). The overexpression of c-Met was significantly more common in poorly differentiated (P < 0.0001) and in the diffusely infiltrated specimens (P < 0.0005). In contrast, TGF beta R II was significantly overexpressed in the well differentiated specimens (P < 0.0001) and associated negatively with c-Met (P < 0.0001). Overall, these data suggest that c-Met/HGF receptor and TGF beta R II overexpression may be involved in the differentiation of human prostatic adenocarcinoma, c-Met with de-differentiation and TGF beta R II with differentiation.

Liquid laboratory waste containing osmic acid and cacodylic acid was mixed with potter's clay or hydraulic cement. The clay-waste product was kneaded into blocks and baked in a klin (1,200-1,400 degrees C). The cement-waste product was allowed to harden into concrete blocks. Some of the baked clay blocks and concrete blocks were ground, and immersed in 1 N NaOH or 10% HCI solutions for 3-6 months. X-ray microanalysis of the dried samples of these solutions showed that no leakage of osmium and arsenic occurred in the baked clay embedding, and that some leakage of these agents occurred in the concrete embedding. The present study indicates that the baked clay embedding method is useful for safe storage of dangerous laboratory wastes. Additional experiments suggested that glass embedding is also useful for safe storage of laboratory wastes or harmful metals.

A sensitive method of HLA-DRB1 typing was devised using a semi-nested polymerase chain reaction (PCR) followed by a restriction fragment length polymorphism (RFLP) analysis (semi-nested PCR-RFLP method). The first-round amplification (30 cycles) of the semi-nested PCR was performed using DRB generic primer pairs and the second round of PCRs (20 cycles) were performed using DRB1 group-specific primers. The products of the second round PCRs were digested with restriction endonucleases for the typing of HLA-DRB1 alleles. By this method, HLA-DRB1 typing was possible from 10 pg of genomic DNA extracted from lymphocytes and from 0.5 microliter of 1,000 times diluted blood without DNA extraction. HLA-DRB1 alleles could be typed from a 2-mm long bloodstained cotton thread prepared from 10 times diluted blood and from a 2-mm thread of whole blood bloodstains stored at room temperature for 2 years. From the mixture of blood of two individuals with different genotypes, DRB1 alleles of the minor component were detected down to 1/1,000 of the major component. This semi-nested PCR-RFLP method is useful for HLA-DRB1 typing from extremely small amounts of DNA and from mixed samples.

POSSUM, a Physiological and Operative Severity Score for the enUmeration of Mortality and morbidity, is a scoring system which assesses perioperative surgical risks (Copeland GP et al.: Br J Surg, 1991, Vol 78, 356-360). The POSSUM scoring system consists of two categories of assessment to assess the risk of surgery. A 12-factor (age, cardiac status, pulse rate, systolic blood pressure, respiratory status, Glasgow Coma Score, serum concentration of urea, potassium and sodium, hemoglobin concentration, white cell count and findings on electrocardiography) and 4-grade physiological score (PS) were developed. This was combined with a 6-factor (type of surgical procedure, number of procedures, blood loss, peritoneal soiling, presence of malignancy and mode of surgery) and 4-grade operative severity score (OSS). The present paper attempts to validate it retrospectively. Postoperative hospitalization period and duration of antibiotics administration were both significantly correlated with OSS, but not with PS. These results suggest that the POSSUM scoring system is useful for predicting the postoperative clinical course.

Conservative treatment is ineffective for ossification of the posterior longitudinal ligament (OPLL) in the thoracic spine, and surgical treatment is indicated for most cases, while such cases are not often experienced. In the present study, the results of surgical management involving mainly posterior decompression for this disease were evaluated clinically. The study included 9 patients (1 man and 8 women) who underwent surgical treatment for OPLL of the thoracic spine between 1984 and 1993. Laminectomy was performed in 5 patients, and laminectomy plus anterior decompression of the OPLL via the posterior approach based on Otsuka's method was performed in 2 patients. In 1 patient, laminoplasty for OPLL of the cervical spine was combined with laminectomy of the symptomatic lesion in the thoracic spine. One patient underwent anterior decompression and fusion. The results were evaluated using the Japanese Orthopaedic Association score (JOA score) and recovery rate. The postoperative follow-up period ranged from 1 year to 10 years and 3 months (mean, 4 years and 6 months). The mean JOA score was 4.8 before surgery and improved to 7.6 at the final examination. This was a mean recovery rate of 50.1%. Symptoms caused by OPLL in the thoracic spine can be alleviated by posterior decompression where OPLL extends from the upper to the middle thoracic spine or extends from the middle to the lower thoracic spine. It seems, however, that OPLL localized to the middle thoracic spine requires anterior decompression.

The ultrastructural and biochemical changes in the brain tissue of 20 mice were studied. The mice, separated into acute and chronic groups, were injected with phorbol myristate acetate (PMA) to trigger the release of free radicals. Malondialdehyde measurement and electron microscopic examination were applied for the evaluation of the effects of the free radicals. The level of lipid peroxide in the chronic PMA group was found to be significantly higher than it was in the acute PMA group (P < 0.005). An electron microscopic examination of the acute group revealed disruption of the mitochondrial cristae and dilatation of the endoplasmic reticulum in the neurons. Myelin sheaths of the nerve fibers exhibited focal structural changes. Neurons and neuroglial cells in the chronic group, however, exhibited distinct ultrastructural alterations. The ultrastructural and biochemical findings showed that free radicals lead to brain damage.

We performed haptoglobin (Hp) genotyping by polymerase chain reaction using allele-specific primer-pairs. The major six genotypes of Hp were identified using this method. Among Japanese individuals living in Ehime and Okayama Prefectures, the allele frequencies were estimated to be Hp2 = 0.723 and Hp1s = 0.277. Genotyping of Hp was possible with 0.3 ng of DNA and with 0.125 microliter of blood. It was also possible with whole blood left at room temperature for a month and also with the bloodstains left at room temperature for three years. In the heated blood samples, both alleles, Hp2 and Hp1s, were detected in those heated at 100 degrees C for 2 h. In bloodstains, Hp2 and Hp1s were detected in samples heated at 100 degrees C for 2 h and 120 degrees C for 30 min. In addition, the genotype could be detected in samples other than blood such as saliva, hair roots, tissue sections and dental pulps. The present method for Hp genotyping is expected to become a useful method in forensic analysis.

Thomsen-Friedenreich antigen (T antigen) has been supposed to be a cancer-specific carbohydrate antigen. We have previously shown that one third of the Japanese population normally expressed T antigen in gastric surface epithelia and the other two thirds expressed fucosyl-T antigen. Their sialylation and blocked-synthesis were associated with diseased conditions. In the present study, we studied gastric surface epithelial expression of monosaccharide antigen Tn, i.e., a precursor of T antigen, and sialyl-Tn. Normal fundic and pyloric epithelia, respectively, expressed Tn supranucleally and cytoplasmically, but did not express sialyl-Tn. In the intestinal metaplasias and intestinal-type adenomas, goblet cells expressed sialyl-Tn in their vacuoles, and absorptive cells expressed Tn apically. In gastric-type adenomas, Tn, but not sialyl-Tn, was detected. Intestinal-type cancers expressed Tn and sialyl-Tn more often than the diffuse-type cancers. Five cancers did not express Tn, sialyl-Tn, or the T-related antigens. In these, four were diffuse-type cancers. We concluded that: a) normal gastric epithelial cells express Tn; b) metaplastic differentiation is associated with sialylation of Tn and c) expression of Tn and sialyl-Tn is depressed in the gastric cancers.