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Using 2-dimensional gel electrophoresis, we previously demonstrated that the S100C protein remarkably decreased after immortalization of normal human fibroblasts, and that this protein caused growth inhibition of human tumor cells when forcibly expressed in these cells, suggesting that S100C plays a significant role in tumor suppression. The present study was carried out to determine what type of human tissues express S100C protein, and, subsequently, whether the S100C content in these tissues changes after normal cells have been transformed into cancer cells. We found that ductal cells in various tissues were positively stained with the S100C protein. In comparison, epithelial cells in digestive organs such as the stomach, small intestine, and colon were not stained as strongly. When 14 pairs of human normal and cancerous tissues were stained with the antibody, decreases in the staining levels of S100C were observed in 6 kinds of cancerous tissues--from the bronchus, mammary duct, renal tubule, prostate, uterus, and testis--in comparison with staining in their normal counterparts. These results suggest that S100C is a new tumor marker protein, the expression of which significantly decreases after malignant transformation of human tissues.

Different bone allografts (pasteurized, autoclaved, and frozen) were compared based on their osteoinductive properties. Our primary purpose was to examine the biologic qualities of pasteurized allografts, as pasteurization inactivates most viruses transmitted by transplantation. Frozen, pasteurized, and autoclaved allografts were packed into a standard defect of rabbit ulna. The animals were sacrificed at 2 and 4 weeks after surgery. The parts of bones with experimental defects were explored en bloc, and a roentgenogram was carried out. Ulna bone samples were then embedded in methyl-methacrylate. Roentgenograms showed that after 2 weeks, calluses were well-formed, but irregular in shape in all 3 types of allografts. After 4 weeks, the calluses were regular in shape in all but the autoclaved grafts. After 2 weeks, the healing processes had begun in the frozen and pasteurized grafts, with the reaching approximately the same stage, while in the autoclaved grafts these processes were not seen and the bone particles were surrounded by connective tissue without any changes. After 4 weeks, osteoinductive processes were very strong, with the first signs of complete bone remodeling at the bone edges of the defect in pasteurized and frozen allografts. The osteoinductive values of these 2 types were very high and similar. Autoclaved allografts, on the other hand, had very low osteoinductive values, as they were still at the very beginning of the healing process. Histomorphometric analysis revealed a significant difference in both newly formed osteoid thickness and osteoblast number per microm of bone surface in all experimental groups (P < 0.005). Values of osteoid thickness and osteoblast number were significantly higher in both frozen and pasteurized grafts when compared with the autoclaved ones (P < 0.005). Osteogenic properties of pasteurized bone allografts were preserved, and the allografts have been gradually replaced with newly formed bone. As such, pasteurized bone grafts from a bone bank have approximately the same biologic validity as frozen grafts, while autoclaved grafts impair bone healing.

We have shown that highly proofreading DNA polymerase is required for the polymerase chain reaction in the genetic analysis of hepatitis C virus (HCV). To clarify the status of HCV quasispecies in hepatic tissue using proofreading DNA polymerase, we performed a genetic analysis of the HCV core protein-encoding region in cancerous and noncancerous lesions derived from 4 patients with hepatocellular carcinoma. In contrast to the previously published data, we observed neither deletions nor stop codons in the analyzed region and no significant difference in the complexity of HCV quasispecies between cancerous and noncancerous lesions. This result suggests that the HCV core gene is never structurally defective in hepatic tissues, including cancerous lesions. However, in 3 of the patients, the consensus HCV species differed between cancerous and noncancerous lesions, suggesting that the predominant replicating HCV species differs between these 2 types of lesions. Moreover, during the course of the study, we obtained several interesting variants possessing a substitution at codon 9 of the core gene, whose substitution has been shown to induce the production of the F protein synthesized by a - 2/+1 ribosomal frameshift.

Postoperative hip alignment was studied on radiographs in cases of total hip arthroplasty (THA) and of Bipolar Head Prosthesis(BHP), both with MX-1. Postoperative anteroposterior-view radiographs of hip joints of patients with a normal hip joint on the unoperated side and without pelvic tilt were used. Thirty-nine THA patients (femoral neck fracture), 26 THA patients (osteonecrosis of the femoral head and osteoarthritis of the hip joint), and 34 BHP patients were selected for this study. Lines and points for measurement of 9 parameters were established on radiographs. The position of the greater trochanter upper edge is 6.5 mm (mean) superior to the femoral head center in the normal hip joint of Japanese, unlike in Caucasians. A femoral head prosthesis should be inserted so that its center and the greater trochanter upper edge are level in order to equalize leg lengths. In BHP cases, the insertion is made so that the greater trochanter upper edge is approximately 4-mm superior to the center of the prosthesis. For further securing of the stem and to equalize leg lengths, stems should be available in 11 diameters from 5-15 mm in 1-mm increments. Postoperative hip alignment in MX-1 THA cases was found to be satisfactory.

We studied parasite detectability in thick films by an acridine orange fluorescence technique (AO) to test its applicability and the use of a Malaria Diagnosis Microscope (MDM)-ESL in the detection of parasites, compared to the conventional Giemsa staining method. This study was conducted on 1,390 clinically suspected malaria cases of Thaton township, Myanmar. We found sensitivities of 82.8% for Plasmodium falciparum (P. falciparum) and 100% for Plasmodium vivax (P. vivax) and specificities of 97.1% for P. falciparum and 98.6% for P. vivax. AO had a higher sensitivity than Giemsa-stained films at low levels of parasitemia (< 1,000/microl). AO showed lower sensitivity and higher specificity than the Giemsa method at parasite levels of more than 1,000/microl. The results of using the AO method, achieved by both novice and experienced observers, showed no significant difference and required less practice to perform the test as well as to identify the parasite. The acridine orange fluorescence technique using a malaria diagnosis microscope MDM-ESL series is simple, rapid and cost effective. The microscope is conveniently operable using standard AC power or a 12-V DC car battery, and it is easily convertible to a conventional biological microscope. With the exception of species differentiation, which is not possible with this method, this method would be appropriate for both clinical and epidemiological studies.

Gastroparesis is a frequent and sometimes life-threatening complication of diabetes mellitus. Autonomic neuropathy seems to be one of the most important mechanisms underlying this entity, together with the other probable pathologies. The present study was performed in order to identify an alternative to gastric scintigraphy as a screening test. The gastric emptying times of 60 subjects (Group 1: 20 insulin-dependent patients, Group 2: 20 non-insulin-dependent diabetes mellitus patients, and Group 3: 20 healthy volunteers) were monitored by gastric scintigraphy. Perception thresholds for cold, heat, and vibration were tested by a quantitative sensory test, and QTc dispersions were calculated from standard electrocardiography recordings. In addition, fasting blood glucose, hemoglobin A1c and urine beta2-microglobulin and microalbumin concentrations were determined for the patient groups. Funduscopic examination was performed by an independent ophthalmologist. Gastroparesis was determined in both patient groups, regardless of fasting blood glucose and hemoglobin A1c concentrations. A strong correlation was observed between nephropathy, retinopathy, and cardiac autonomic denervation (QTc) and gastroparesis. In conclusion, retinal and renal microvasculopathy parameters and cardiac autonomic function tests may be useful for screening diabetic patients for gastroparesis.

We describe a modified method for typing a polymorphic microsatellite D12S391 locus by PCR using a newly designed primer pair. This primer pair produces shorter D12S391 amplified fragments (104-156 bp) than the primer pair originally described by Lareu et al. (209-261 bp). The detection system for the D12S391 locus using the new primer pair and capillary electrophoresis (CE) analysis was evaluated using various forensic samples. The typing results from 70 DNA samples using the new primer pair and the original primer pair were completely identical. One hundred twenty-five amplified fragments from D12S391 alleles were sized correctly within +/- 0.25 bp of the D12S391 allelic ladder. A rare allele, 19.3, previously found only in Caucasians, was found for the first time in a Japanese subject, and it was clearly distinguished from allele 20 by the CE analysis. This detection system was sensitive and could detect D12S391 types from 16 pg of genomic DNA, and from a minor component at a ratio of 1:10 in mixed samples. This system was more useful for the analysis of degraded DNA than was the method using the original primer pair, and could detect D12S391 types from bloodstains that had been stored for 26 years. In addition, the specificity of the method was demonstrated using nonhuman DNA.

Genetic and molecular biological methodologies are being applied to the study of patients with epilepsy at an ever-increasing pace. Accurate classification of epilepsy within large families has allowed identification of genes through linkage analysis and then isolation of gene products. Mutations causing ion channel abnormalities coupled with clinical patterns of focal epilepsy syndromes are beginning to change our thinking about the etiology of recurrent seizures in all patients. Molecular methodology is beginning to have impact on understanding of the mechanisms of actions of drugs used to treat epilepsy and will have an impact on how future treatments are designed.

This study involved the examination of 1,006 chest x-ray films of workers from the industries devoted to shipyard welding, stone grinding, and refractory crushing in southern Okayama prefecture. Of the reviewed films, analysis was focused on subjects with a profusion rate of 0/1 as well as pneumoconiotic subjects (exhibiting profusion rates of 1/0 or greater) in order to discover cases in the beginning stages. One-hundred-and-seventy-four films illustrated a profusion rate of 0/1 or greater, and the proportion of this profusion rate was revealed to be highest in shipyard welders. Even some workers under 40 years of age were found to have already developed pneumoconiosis. Of these 1,006 subjects, 30 volunteers permitted us to measure their personal dust exposure concentrations. The measured concentration of the shipyard welders' dust exposure (respirable dust; 3.3 86.3 mg/m3, total dust; 7.5-117.0 mg/m3) was higher than those of the other 2 industries. Statistical differences among the industries were observed in the respirable dust concentrations. A statistically significant positive correlation was demonstrated between the working duration in dusty environments and the rate of profusion. The present findings suggest the need for taking adequate measures in Okayama in order to prevent workers from developing, or to help retard the progression of, pneumoconiosis.

Tremendous excitement has been generated by the use of botulinum toxin for the treatment of various types of urethral and bladder dysfunction over the past several years. Botulinum toxin is the most lethal naturally occurring toxin known to mankind. Why, then, would an urologist want to use this agent to poison the bladder or urethral sphincter? In this review article we will examine the mechanisms underlying the effects of botulinum toxin treatment. We will discuss the current use of this agent within the urologic community and will provide perspectives on future targets of botulinum toxin.

In this study, we tested brain surface cooling as a new method of inducing selective brain hypothermia, and evaluated its effects on focal cerebral ischemia using a cat model of transient middle cerebral artery (MCA) occlusion. Cats underwent 1 h of MCA occlusion followed by 5 h of reperfusion. Brain surface cooling was induced for 4 h during and after MCA occlusion in the hypothermia group, but not in the normothermia group. Brain surface cooling was performed using saline perfusion into the subdural space. Rectal temperature, brain surface temperature, and deep brain temperature were monitored, and regional cerebral blood flow (rCBF) and somatosensory evoked potential (SEP) were serially measured. After 5 h of reperfusion, water content was also measured. Although the rectal temperature was maintained at about 37 degrees C, the brain surface temperature decreased rapidly to 33 degrees C and was maintained at that temperature. For 3 h following reperfusion, the rCBF was lower in the hypothermia group than in the normothermia group. At 4 and 5 h after reperfusion, the recovery of SEP amplitude was significantly more enhanced in the hypothermia group than in the normothermia group. In the gray matter, the water content was significantly more diminished in the hypothermia group than in the normothermia group. These results demonstrate that our method is useful for protecting the ischemic brain from a transient MCA occlusion. This method may be adapted for neurological surgery.

Transforming growth factor-beta1 (TGF-beta1) exerts potent immunosuppressive effects. In this study, we investigated the potential role of TGF-beta1 produced by hepatocellular carcinoma (HCC) cell lines in immunosuppression mechanisms. Using the Mv1Lu cell-growth inhibition assay and an enzyme-linked immunosorbent assay (ELISA), we detected optimal levels of TGF-beta1 in the culture supernatants conditioned by the HCC cell lines PLC/PRF/5, Hep3B, and HepG2. To determine the biological activity of TGF-beta1 in the supernatants, we examined the effects of the culture supernatants on the production of interferon (IFN)-gamma induced during the culture of peripheral blood mononuclear cells (PBMCs) stimulated with interleukin (IL)-12. IFN-gamma production of IL-12-stimulated PBMCs in the 1:1 dilution of the acid-activated conditioned medium of PLC/PRF/5, Hep3B, and HepG2 reduced to 14.7 +/- 0.8, 17.3 +/- 9.0, and 35.9 +/- 14.6%, respectively, compared with the value in the culture with control medium (complete culture medium). These results suggest that HCC cells producing TGF-beta1 may reduce the generation or activation of cytotoxic T lymphocytes (CTL) and natural killer (NK) cells, and thus could enhance their ability to escape immune-mediated surveillance.

We have previously developed an enzyme-linked immunosorbent assay (ELISA) to measure stool decay-accelerating factor (DAF) and found that stool DAF concentrations were significantly elevated in patients with colorectal cancer, suggesting that the measurement of stool DAF may be valuable for the detection of colorectal cancer. In order to refine the assay for the measurement of stool DAF, we investigated 1) effects of centrifugation of stool samples, 2) effects of detergents, and 3) adequate combination of various anti-DAF monoclonal antibodies for the ELISA system using only monoclonal antibodies. We found that high-speed centrifugation could be omitted and that only the removal of large undigested food residues by centrifugation of short duration in a low-speed benchtop microcentrifuge sufficed to adequately prepare the stool samples. Addition of 2 detergents, octyl beta-glucoside and sodium deoxycholate, known to solubilize glycosyl-phosphatidylinositol-anchored proteins such as DAF, did not influence stool DAF values. By using 2 mouse anti-DAF monoclonal antibodies (clone 4F11 and 1C6), we were able to achieve a stable ELISA for the measurement of stool DAF using a uniform source of antibodies. The results should allow us to consistently apply the DAF assay for routine use in the detection of colorectal cancer.

We have designed to apply 2-pyridine aldoxime methiodide(PAM), considered to be a cholinesterase reactivator in vitro, both to the laboratory rabbits poisoned by parathion and to the patients of parathion poisoning, and obtained the following results: 1. With administration of PAM, a prompt and complete dispersion of symptoms of the poisoning can be realized. 2. Cholinesterase activity of red blood cell has instantly and completely recovered, and that of serum transiently. 3. The amount of serum mucoprotein and the activity of active protein-SH-radical of serum varied in direct proportion to the activity of serum cholinesterase. 4. Generally, an intravenous injection of 1g. PAM is sufficient even in the severe case and it may be increased when necessary. 5. The ill effect has not been encountered in the PAM administration. 6. PAM exerts no influence on the cholinesterase activity of normal blood. 7. PAM is expected to play an important role as a prophylactic agent of alkylphosphate poisoning. From these results it seems clear that PAM is a specific and effective antidote against alkylphosphate intoxication.

To reveal the relationship between the morphologic structure and the functioning of cells it is of great importance to know the molecular arrangement in cytoplasm. For this purpose the observation by polarization microscope is one of the most exellent methods. There are several excellent works on nerve fibre done using polarized lightl,2,3,4 but there is no report concerning the nerve cell itself. It is the purpose of this paper to show our data obtained on ganglion cells and nerve fibres with the deduced speculation on the structural arrangement of lipoprotein in the protoplasm of nerve cells and fibres.

The existence of autonomic adjustment functions of eye pressure was demonstrated in various ways, both clinically and experimentally. It is possible to consider that in normal condition, I.O.P. is controlled autonomically like cardiac or respiratory rate irrespective of the internal or external influences of the body. The auther calls such a phenomenon "autonomic eye pressure adjustment function". The mechanism of this physiological function will be reported on in articles to follow.

1. The shell of the ova of Enterobius vermicularis is composed of two chitinous layers, a compact outer layer and a looser inner layer. Both surfaces of the two layers have a dense border. 2. The outer layer has innumerable tubules about 0.2 micron in diameter, and only those tubules found in the thinner part of the layer open to the outside and inside. 3. The outer layer of mature eggs is about 0.45 micron and the inner layer about 2 to 6 microns, in thickness. 4. The chitinous microfibrils in both layers form a rcticular structure which contains very fine granules in the mesh.

In 1955 SANO found mitochondria by the supravital stain with Janus green B in the basophilic stippled cells from the circulating blood of the lead intoxicated rabbitsl , and in 1956 by means of electronmicroscope VALLEJO-FREIRE, BRUNNER et al. found mitochondria in the reticulocytes2,3, and later at the end of 1956 BRAUNSTEINER et al. also succeeded in revealing mitochondria and the vesicular structure by electron microscope in the ultra thin section of young red cells4. We also have found the mitochondria and the endoplasmic reticulum in young red cells. It has been discussed long whether the reticulum of reticulocytes is a preexistent structure or an artifact. The fact that the mitochondria exist in the reticulocyte seems to support strongly the preexistence theory of the reticulum, substantia reticulo filamentosa. However, the fact that the reticulum has several characteristics different from the general mitochondria5,6 can not be ignored. In this paper we should like to demonstrate the photos of mitochondria and the endoplasmic reticulum in the denucleated red cells revealed by electron microscope comparing to the picture of reticuluocyte appeared by supravital stain.

The evaluations of the basal metabolism before the operation, at the completion, and at the follow-up examinations, have been conducted on the patients with cervical carcinoma, all under the age of 50 years, and operated on in the Clinics of Obstetrics and Gynecology, Okayama University Medical School and the following results are obtained: 1. The basal metabolism of the patients with cervical carcinoma of either stage I or stage II as compared with that of the normal women is more accelerated; while the basal metabolism of the cervical carcinoma of stage II. is more augmented than that of the cervical carcinoma stage I. 2. The basal metabolism at the completion of operation is markedly lower than that before the operation. 3. In those who received the autotransplantation of the ovary concomitantly with operation, the basal metabolism once increases at the 4th month after the operation and after that it maintains a low stable state. 4. The basal metabolism of those who received the operative castration continues increasing up to the 8th month after the operation and thereafter it returns to a rather balanced state. 5. From the aspects of the fluctuations of the basal metabolism, it has been recognized that the autotransplantation as compared with the castration exerts less influences on the somatic system and for a shorter period of time, and also the endocrine system of the former returns to the balanced condition earlier.

1. Adversive movements were induced by electrical stimulation and metrazol injection on area 4c of the cerebral cortex. 2. The adversive movement from area 4c does not pass through the thalamus, nucleus caudatus, nucleus lenticularis or superior collicuIus, but through direct efferent pathways in the internal capsule. 3. The adversive movement from area 4c passes through the pyramidal tract.