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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.

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.

Origins and distribution of the human inferior phrenic arteries were studied by dissecting 68 Japanese adult cadavers. The inferior phrenic arteries were usually observed as paired (left and right) vessels. Their origins were summarized as follows: a) the aorta itself (85/138 cases, 61.6%), b) the ventro-visceral arteries (celiaco-mesenteric system of the aorta) including the celiac trunk (39/138 cases, 28.2%) and the left gastric artery (4/138 cases, 2.9%), and c) the latero-visceral arteries (adreno-renal system of the aorta) including the middle adrenal artery (4/138 cases, 2.9%) and the renal artery (6/138 cases, 4.3%). The left and right arteries occasionally originated in common trunk from the aorta, celiaco-mesenteric system or adreno-renal system (22/138 cases, 15.9%). A typological diagram explaining these variations is given. The inferior phrenic arteries, especially the left ones, sometimes issued visceral or esophageal branches. This fact indicates that the inferior phrenic arteries are homologous with the celiac trunk and mesenteric arteries. It is further discussed that the celiac trunk and mesenteric arteries are originally paired vessels, through introduction of our previous typological diagram of the abdominal arteries.

Recent data suggested that platelet-activating factor (PAF) could play a pathophysiologically important role in the progression of hypoxic-ischemic brain injury. We investigated brain tissue PAF concentration in the hypoxic-ischemic brain of immature rats. Endogenous PAF concentration in brain tissue showed a marked increase in hypoxic-ischemic pups (Group 1, 85.6 +/- 15.5 pg/mg protein) when compared to that of the control (9.1 +/- 3.1 pg/mg protein). In addition, we studied the effects of pretreatment with L-carnitine (5 days and 2 h before the hypoxia) on endogenous PAF concentration in the hypoxic-ischemic brain. Endogenous PAF concentration in the short-term pretreatment group (Group 2, 81.6 +/- 9.7 pg/mg protein) was not different than in Group 1 rat pups. However, a significantly decreased PAF concentration was found in the group of pups that received carnitine pretreatment for 5 days (Group 3, 30.5 +/- 11.0 pg/mg protein). These results indicate that PAF is an important mediator in the immature rat model of cerebral hypoxic-ischemic injury. The suppressor effect of L-carnitine on PAF production may give new insight into the treatment of hypoxic-ischemic brain injury.

We studied the association of fatty liver with subcutaneous and visceral obesity in 46 male and 36 female patients with body mass index (BMI) over 22 kg/m2. The correlation coefficient between the ratio of the visceral adipose tissue to the subcutaneous adipose tissue (V/S) and the computed tomography (CT) number of the liver was -0.299 (P < 0.05) and that between the V/S ratio and the ratio of the CT number of the liver to that of the spleen (CT-L/CT-S) was -0.335 (P < 0.05) in the males. Partial correlation analysis after making correction for BMI showed an increased correlation coefficient of -0.485 (P < 0.05) between the V/S ratio and the CT-L/CT-S ratio in the males. The odds ratio in the males for CT-L/CT-S below 1.0 and V/S above 1.0 was 3.25 with a 95% confidence interval of 1.02 to 9.39. No such association between the V/S ratio and the CT-L/CT-S ratio was present in the female patients. Multiple regression analysis with serum level of alanine aminotransferase, a marker of fatty liver, as an independent variable revealed a partial regression coefficient of -17.7 for CT-L/CT-S (P < 0.05) in the males and -21.7 (P < 0.05) in the females, validating the CT-L/CT-S ratio as an index of fatty liver. The results indicate the association of fatty liver as determined by the CT-L/CT-S ratio with visceral obesity in males.

We studied the magnetic resonance imaging (MRI) of 120 knees in 86 rheumatoid arthritis (RA) patients and of 14 unaffected knees in 12 control cases. We also developed a scoring system as a quantitative analysis method. We divided the MRI into 10 items, and classified the severity of the symptoms into 4 grades (score 0 to 3). The average total score increased according to the radiographic grade. Soft tissue lesions were clearly detected, even in the early stages of RA. Items such as synovial proliferation showed a high score even in the early stages, suggesting that it was the initial symptom of RA. The score also showed a correlation with the inflammatory signs. These results suggest that this scoring system is very sensitive and yields a good reflection of RA activity. We demonstrated that this system is simple and convenient for routine diagnostic use. We further demonstrated that it is useful for following the advancement of RA and for evaluating the response to treatment.

Thirteen patients exhibited a communicating hydrocephalus following subarachnoid hemorrhage secondary to ruptured intracranial aneurysms and were treated with shunt procedures. The interval between subarachnoid hemorrhage and surgery averaged 9 weeks. Seven of the patients showed improvement. The prognostic value for surgical management was evaluated on the basis of three different diagnostic examinations (computed tomography(CT), cisternography and constant infusion test). A correct diagnosis was obtained in 78 per cent in cisternography, and 63 per cent in infusion test and CT. All patients responding to surgery showed a typical pattern in cisternography, consisting of ventricular retention of radiopharmaceutical tracer for 48 h or longer in association with no radioactivity over the cerebral hemispheres. The constant infusion test correlated well with typical cisternographic patterns. CT is useful in demonstrating pathophysiological changes in hydrocephalus. Periventricular hypodensity was visible in patients with normal or slightly elevated intracranial pressure, accompanied by fairly rapid deterioration. All of them responded well to shunting. In most cases which benefited from the shunt, the postoperative CT showed not only normal-sized ventricles but also marked regression of the hypodensity over a short period.

The distribution of corticotropin releasing factor (CRF) and arginine vasopressin (AVP) in hypothalamic nuclei were examined in control and estrogen-treated female rats. CRF activity was measured using monolayer cultured rat anterior pituitary cells and AVP by radioimmunoassay. Hypothalamic nuclei were punched out according to the method of Palkovits. The distribution of CRF activity in 5 different hypothalamic nuclei was similar to that of AVP in intact female rats. CRF activity in hypothalamic nuclei, pituitary ACTH content and plasma ACTH levels in estrogen-treated rats were not significantly different from those in control rats. However, significant elevation of AVP content was observed in the supraoptic and paraventricular nuclei of estrogen-treated rats. These results indicate that CRF and AVP are distributed in similar hypothalamic nuclei, but that they are not identical.

A perifusion method has been developed using rat hypothalamo-neurohypophyseal system (HNS) or neural lobe to investigate the control mechanism of arginine vasopressin (AVP) release. A specific radioimmunoassay (RIA) for AVP was developed to measure AVP in perifusion medium employing anti-AVP serum which was obtained by immunizing rabbits. At a final dilution of 1/12,000, the antiserum showed less than 0.66 and 0.01% cross reactivity with lysine-vasopressin and oxytocin, respectively. But it did not cross reacted with other peptide hormones. The lowest detectable level of vasopressin was 0.5 pg/tube. The intra-assay coefficient of variation averaged 10.4%. The dilution curve of perifused medium was well paralled to the standard curve of AVP assay. AVP release from HNS or neural lobe gradually declined to the stable level in 90-120 min after the initiation of perifusion. Good repeatability of the AVP release from neural lobe was recognized by repeated stimulation with 10 min perifusion of 60 mM KCl at every 60 min. HNS released AVP in dose related manner to the osmotic challenge of sodium or glucose, and AVP release was stimulated from HNS by prostaglandin E2, but not by dopamine. These results show that the perifusion methods using AVP-RIA is a useful method to examine the AVP release from HNS or neural lobe.

Chromosome studies were conducted on two patients with adult T-cell leukemia. In both patients, a marker chromosome 14q+ and a structural change involving chromosome 1 with trisomy of the q arm were found in peripheral blood leukocytes. The 14q+ marker chromosome had resulted from translocation from #5p in one patient and #5q in the other patient. The present and previous studies suggest that the donor chromosomes involved in the 14q+ translocation are variable. This indicates that the 14q+ marker chromosome rather than the donor chromosome is intimately related with adult T-cell leukemia.

The distribution of corticotropin releasing factor (CRF) and arginine vasopressin (AVP) in hypothalamic nuclei were examined in control and estrogen-treated female rats. CRF activity was measured using monolayer cultured rat anterior pituitary cells and AVP by radioimmunoassay. Hypothalamic nuclei were punched out according to the method of Palkovits. The distribution of CRF activity in 5 different hypothalamic nuclei was similar to that of AVP in intact female rats. CRF activity in hypothalamic nuclei, pituitary ACTH content and plasma ACTH levels in estrogen-treated rats were not significantly different from those in control rats. However, significant elevation of AVP content was observed in the supraoptic and paraventricular nuclei of estrogen-treated rats. These results indicate that CRF and AVP are distributed in similar hypothalamic nuclei, but that they are not identical.

In order to elucidate the specific thyrotropic area in the hypothalamus, thyrotropin releasing hormone (TRH) content and concentration were measured in discrete hypothalamic nuclei and areas after triiodothyronine (T3) administration (T3 10 micrograms/rat/day for 6 days), thyroidectomy (TX) and acute cold exposure in male rats. In th TX and T3 groups, serum TSH levels were significantly increased in TX group and markedly decreased in T3 and TX with T3 groups as compared to the sham operated control group (Sham). TX produced a slight but nonsignificant decrease in TRH content in most of the hypothalamic nuclei examined as compared with the Sham group. However, a significant increase in TRH contents was seen in the anterior hypothalamic nucleus (AHN), median eminence (ME) and posterior pituitary (PP) in TX with T3 group as compared to the rats with only TX. In the acute cold stress experiments, serum TSH levels were elevated from 15 to 30 min of 4 degrees C exposure. Together with these peripheral changes, TRH content and concentration in the suprachiasmatic nucleus (SC) were increased significantly at 15 min and had returned to the normal level by 30 min after 4 degrees C cold exposure. However, in the paraventricular nucleus (PV) and dorsal premammillary nucleus (PMD), marked decrease in TRH concentrations were observed with this stress. Therefore, 1) decreased TSH release in TX rats treated with T3 was induced by the block of TRH release from the AHN and ME as compared with the TX group, and 2) elevated serum TSH levels in 4 degrees C cold stress might be induced by the release of TRH from the PMD and PV. These experiments demonstrate that the specific hypothalamic area for TSH release was located in some of the anterior and posterior hypothalamic nuclei and in the ME.

The effect of cytochalasin B (CB) on the surface structure of Ehrlich ascites tumor cells was investigated using the scanning electron microscope. The effect occurs in two steps: formation of zeiotic knobs on the cell surface and subsequent grouping of the knobs at one pole of the cell. The early step of zeiotic knob formation occurs at low concentrations of CB (0.5-1 microgram/ml) at 37 degrees C and at high concentrations of the drug (5-10 microgram/ml) at low temperature but within 1 min at 37 degrees C. This step is only partially inhibited by 5 x 10(-3) M sodium azide. The subsequent grouping of zeiotic knobs lasts for more than 2 min at 37 degrees C and occurs only in the case of high concentrations of CB. It is inhibited by sodium azide and is often associated with grouping of the microvilli, which are then lost from all of the cell surface except the area of knob-grouping.

3-(3-methyl-3-butenyl)-1,2,3,4,5,6-hexahydro-6, 11-dimethyl-8-hydroxy-2,6-methano-3-benzazocine (KF-1820) is a derivative of benzomorphan and is different from pentazocine only in the site of the double bond. KF-1820 showed potent analgesic activity in all tests performed. KF-1820 was 6 to 12 times and 30 to 40 times more potent than morphine and pentazocine, respectively, when administered subcutaneously. KF-1820 had little or no narcotic antagonist property. Physical dependence liability tests indicated that KF-1820 may be a little less, or as liable as, pentazocine to produce physical dependence. ID50 values of KF-1820, pentazocine and morphine for depression of contractions of isolated guinea pig ileum to coaxial stimulation correlated well with their analgesic activities in the rodent. The dissociation equilibrium constant of KP-1820 confirmed the in vivo finding that KF-1820 had little or no narcotic antagonist property.