Organic analysis. XLIV. A new deproteinizing agent for determination of blood sugar.
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Biomedical subjects
Publications and source records attributed to K Ohashi.
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As a regional therapy for hepatic malignancy, we developed a simple method of isolated liver perfusion (hyperthermo-chemo-hypoxic). In the present study, the influence of this method on the hepatic tissue and other organs was experimentally evaluated and applied it to seven patients. Experimentally, all dogs survived without hepatic insufficiency and systemic toxicity. Clinically, one patient died on postoperative day 14 of hepatic failure. The reason was that liver temperature reached 43 degrees C, which seemed to be the maximum limit for thermal toxic effect to the human liver. The other six patients well tolerated the perfusion with mild increases of serum aminotransferase and total bilirubin levels and decreases of hepaprastin levels. Serum aminotransferase and total bilirubin and hepaprastin levels returned to normal levels by postoperative day 14. There were no significant differences between the isolated liver perfusion group (n = 7) and hepatectomy-only group (n = 27). Six patients were disease-free during the observation period after the perfusion. This system is a simple, useful method for treating patients with metastatic cancer limited to the liver.
The trophoblast cells which contact with maternal tissue express HLA class I either weakly or not at all. This phenomenon is beneficial for the protection of the fetus from immunological rejection by the mother. However, the mechanisms of reduced expression of HLA class I are unexplained. Therefore, we utilized choriocarcinoma cells to analyse the suppressive mechanisms to HLA class I expression in trophoblasts. In this paper we reported the heterogeneity of HLA class I expression in five choriocarcinoma cell lines. The levels of surface HLA class I molecules correlate approximately with mRNA of HLA class I heavy chain, and all choriocarcinoma cells produce moderate amounts of surface and secreted beta 2 microglobulin. It was also found that interferon gamma upregulates HLA class I molecules and mRNA in two choriocarcinoma cell lines, NUC1 and HCCM5.
We investigated the effect of the activation of complement in human serum on isolated adult porcine islets using an in vitro model of pig-to-human islet transplantation. Pancreata were obtained from slaughterhouse pigs (6-8 mo old). Islets were prepared by intraductal collagenase digestion followed by purification on Ficoll gradients. The purified islets were incubated with xenogeneic human serum with or without heat inactivation for 45 min. As control, islets were incubated with autologous porcine serum. After the incubation, the islets' responsiveness to an acute glucose stimulus (5.5 mM, static incubation) was evaluated by measurement of the insulin content of the medium. Islets exposed to human serum showed significantly lower insulin secretory response than the control (1.76 +/- 1.17 microU/islet/120 min, without heat inactivation; 1.74 +/- 1.36 microU/islet/120 min, with heat inactivation; 3.39 +/- 0.92 microU/islet/120 min, control). No difference in insulin secretory response, however, was observed between islets exposed to human serum with heat inactivation and without. Furthermore, we evaluated the cytotoxic activity of human serum on porcine islets by a complement-dependent cytotoxicity using the MTT colorimetric assay, and found that the human serum had no complement-dependent cytotoxic activity against the islets. We concluded that the insulin secretion dysfunction of porcine islets exposed to human serum was not due to the activation of complement and there was no evidence of hyperacute rejection mediated by complement activation in the in vitro model of pig-to-human islet transplantation.
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To assist in the development of new approach to the palliation and treatment of intrahepatic cholangiocarcinomas, we classified tumours into mass-forming (MF), peri-ductal extension (PD), and spicula-forming (SF) types in 14 subjects who underwent surgical treatment. Lymph node metastasis and microscopic lymphatic invasion were pronounced in the PD and SF types. Furthermore, in SF type tumours the incidence of microscopic vascular and perineural invasion was high. The proliferating cell nuclear antigen labelling index, a reflection of the proliferation rate of tumour cells, was significantly higher in PD and SF types than in the MF type. The prognosis associated with the MF type tended to be better than that of the other two types. For the MF type, liver resection with tumour free margins must be performed, whereas for the PD and SF types, aggressive treatment, that is a combination of extensive liver resection, lymph node dissection, and effective adjuvant anti-cancer therapy should be provided to aim at life-long cure.
We describe a case of primary localized nodular tongue amyloidosis associated with Sjögren's syndrome in a 62-year-old woman. The presence of Sjögren's syndrome was confirmed both serologically and histologically. The amyloid tumor, which was marginally excised, recurred 3 years later and was re-excised. Immunohistochemical examination revealed that the amyloid protein was of the AL (lambda-light chain) type. Infiltration of plasma cells was observed around the minor salivary glands of the tongue surrounded by amyloid. The relationship between the plasma cells and amyloid deposition is discussed.
The serum insulin, fractional absorption, serum human C-peptide, and plasma glucose responses of normal fasting subjects were compared after the subcutaneous and intravenous administration of human insulin (recombinant DNA) and Novo Actrapid insulin. While no statistical difference at each time point was observed between the two insulins, for each time point after 0.05 and 0.1 U/kg s.c., the 0-6 h area under curve (AUC) after the 0.05 U/kg dose was greater for human insulin than for pork insulin. There was no difference in the 0-6 h AUC after the 0.1 U/kg s.c. dose. The serum human C-peptide responses to the two insulins were virtually identical. With the 0.05 U/kg s.c. dose, hypoglycemic effect of human insulin was greater than for Actrapid. This difference did not occur after 0.1 U/kg s.c. Following intravenous administration using 0.05 U/kg, the serum IRI, serum C-peptide, and glucose responses were the same. These data indicate only slight differences between human insulin and Actrapid insulin.
In a series of animal experiments on guinea pigs and rats, conducted in an attempt to find a better solution for the treatment of sensorineural hearing impairment and tinnitus-known as diseases almost impossible to heal-it was revealed electrophysiologically, biochemically and electronmicroscopically that loop diuretics, referred to as ototoxic drugs, cause an increased amount of the therapeutic preparation to transfer to the inner ear fluid. It is concluded, on the basis of the encouraging results of these experiments, that combined medication with metabolic activator, steroid hormone and loop diuretics is significantly more effective in improving the symptoms of patients with inner ear deafness and tinnitus.
To measure the endolymphatic and perilymphatic spaces, we used human temporal bones (horizontal serial sections) under two selection criteria: absence of otological pathology, and absence of artifact in the membranous labyrinth (boundary between endolymphatic and perilymphatic spaces) maintaining an intact structural integrity. Under magnified projection, the area of scala tympani, scala vestibuli, scala media, vestibular endolymphatic space, and vestibular perilymphatic space were measured separately, using a microcomputer digitizing tablet. Three repeated measurements were obtained and averaged. The mean total labyrinthine fluid space was 204.5 mm3; the mean total endolymphatic space was 38.1 mm3 and mean total perilymphatic space 166.4 mm3. The mean total vestibular fluid space was 120.9 mm3 and mean total cochlear fluid space 83.6 mm3. In the vestibule, the perilymphatic space occupied 74.8%, and the endolymphatic space, 25.2%, whereas 90.8% of the cochlear fluid space was occupied by perilymph.
To characterize the molecular events involved in both apoptosis and transformation process induced by Marek's disease virus (MDV), the expressions of the bcl-2 and bcl-x genes, ones of the dominant apoptosis-regulating genes, in Marek's disease (MD) tumor cell lines and cells prepared from MDV-infected chickens were analyzed. The expression of bcl-2 was down-regulated in both CD4+ and CD8+ T cells prepared from MDV-infected chickens at 3 weeks p.i. No bcl-2 transcript was detected in MD tumor-derived MSB1 and MTB1 cell lines, which had been established from primary MD tumors. On the other hand, the bcl-xL transcript whose product can also inhibit apoptosis was expressed in cell lines derived from MD. By the treatment with phorbol 12-myristate 13-acetate (PMA) and ionomycin, normal CD4+ T cells were induced to express bcl-xS which can promote apoptosis, while bcl-xL was constitutively expressed in MD cell lines. Our results suggest that bcl-xL rather than bcl-2 might play an important role in the transformation process by MDV.
BACKGROUND/AIMS: Intraductal papillary mucinous tumors of the pancreas have been recognized as a distinct clinical entity. However, their biological behavior has not been clearly defined. The aim of this study was to examine the prognosis of this tumor, to clarify the biological behavior and determine the most appropriate treatment. METHODOLOGY: Correlations between prognosis of operated cases and histopathologic features were investigated. RESULTS: In 105 patients with characteristic clinical features of intraductal papillary mucinous tumors, the lesions were classified as hyperplasias in 21%, intraductal tumors in 48% and invasive carcinomas in 31%. Minimal invasion was apparent in 25%, lymph node metastasis in 21%, and fistula formation in 31% of the invasive lesions. Non-invasive and minimally invasive intraductal papillary mucinous tumors were essentially free from risk of tumor recurrence. Other invasive intraductal papillary mucinous tumors showed a significantly poor prognosis. CONCLUSIONS: Because of the variation in pathological characteristics, patient outcome and the possibility of differential diagnosis, the treatment might be recommended as follows: the case of hyperplasia can be followed-up with close surveillance. Non-invasive and minimally invasive intraductal papillary mucinous tumors should be operated with function-preserving minimal pancreatectomy. For patients with invasive intraductal papillary mucinous tumors evident with preoperative imaging modalities, radical operations with lymph node dissection might be needed.
BACKGROUND/AIMS: IPMT (intraductal papillary-mucinous tumor) of the pancreas has unique clinicopathological characteristics. The lesions which show characteristic clinical features of IPMT exhibit a wide spectrum of histological types ranging from atypical hyperplasia to invasive cancer. Therefore, surgical treatment cannot be recommended for all patients with IPMT. It is necessary to assess the malignant potential of IPMT in individual patients in order to select an appropriate approach. The aim of this study was to evaluate the effectiveness of endoscopic ultrasonography and intraductal ultrasonography as compared with ultrasonography and computed tomography for this purpose. METHODOLOGY: Ultrasonography, computed tomography, endoscopic ultrasonography and intraductal ultrasonography were performed in 49 cases of IPMT (atypical hyperplasia 7, adenoma 23, noninvasive 7 and invasive adenocarcinoma 12). On the basis of the histopathological analysis of another 28 cases of resected IPMT specimens, criteria for differential diagnosis by imaging modalities were defined as follows: Nonneoplastic lesion (atypical hyperplasia): no wall thickening or nodule; noninvasive IPMT (adenoma and intraductal carcinoma): a nodule or wall thickening is present; and invasive IPMT with pancreatic parenchymal invasion: a mass with a heterogenous pattern or interruption of the pancreatic duct wall by the mass. RESULTS: The diagnostic accuracy rate for differentiating nonneoplastic lesion noninvasive IPMT, and invasive IPMT was 33% by ultrasonography, 38% by computed tomography, 77% by endoscopic ultrasonography, and 67% by intraductal ultrasonography. Sensitivity, specificity and accuracy rates for differentiating neoplastic and nonneoplastic IPMT by ultrasonography was 33%, 100%, 42%, by computed tomography 36%, 100%, 44%, by endoscopic ultrasonography 90%, 71%, 88%, by intraductal ultrasonography 94%, 29%, 84%, respectively. Sensitivity, specificity and accuracy rates for differentiating invasive and noninvasive IPMT by ultrasonography was 25%, 100%, 80%, by computed tomography 33%, 100%, 83%, by endoscopic ultrasonography 55%, 97%, 88%, by intraductal ultrasonography 56%, 91%, 84%, respectively. Diagnostic accuracy for invasive IPMT except minimally invasive cases by endoscopic ultrasonography and intraductal ultrasonography was 80%, based on the results of the examination which demonstrated a higher grade lesion. CONCLUSIONS: With these criteria, ultrasonography and computed tomography showed high specificity, but low sensitivity for the differential diagnosis of neoplastic/nonneoplastic and invasive/noninvasive IPMT. However, endoscopic ultrasonography and intraductal ultrasonography had high sensitivity and diagnostic accuracy for the differential diagnosis of neoplastic/nonneoplastic lesions. Combination of endoscopic ultrasonography and intraductal ultrasonography showed a high accuracy rate in the diagnosis of invasive IPMT. Thus endoscopic ultrasonography and intraductal ultrasonography contributed significantly to the choice of the treatment for IPMT.
Techniques of plasma filtration with rotating flat membranes are capable of producing a Taylor vortex and concomitantly reducing concentration polarization by formed elements. The present study describes filtration characteristics of a small-sized rotating filter that is superior to a capillary filter in terms of filtrate flux. The rotating filter with a surface area of 60 cm2 was composed of two concentric cylinders, with the inner cylinder mobile, with a flat polycarbonate membrane (pore diameter: 0.8 micron), and the outer cylinder fixed. Filtration experiments were performed using bovine blood to determine filtrate flux at rotational speeds from 600 to 4,500 rpm. Photographic observations using aluminum powder revealed that many Taylor vortices were present in the radial gap (0.6 mm). Filtrate flux increased with transmembrane pressure until a maximum filtrate flux of 0.02 cm/min at 600 rpm, 0.42 cm/min at 1,800 rpm, 0.60 cm/min at 3,600 rpm, and 0.69 cm/min at 4,500 rpm were reached at a blood flow rate of 100 ml/min. Based on the filtration resistance model, values for fractional filtration resistance of the red blood cell polarization layer ranged from 0.25 to 0.80 at a rotational speed of 3,600 rpm, whereas those of a capillary filter range from 0.80 to 0.90. This indicates that Taylor vortices promote back diffusion of red blood cells accumulated on membrane surfaces. Thus, the filtrate flux of a rotating filter is two orders of magnitude higher than that of a capillary filter because of the back diffusion of red blood cells facilitated by Taylor vortices.
The pharmacokinetics and pharmacologic effects of human insulin (Novo) derived from porcine insulin were studied by a double-blind crossover comparison with porcine monocomponent insulin in healthy volunteers. Both insulins were given by subcutaneous (s.c.) and intravenous (i.v.) injection to healthy male volunteers. The doses of injected insulin were 0.05 U/kg and 0.1 U/kg for the s.c. study, and 0.025 U/kg and 0.05 U/kg for the i.v. study. The plasma immunoreactive insulin (IRI) increased rapidly, plasma C-peptide and glucose decreased gradually, and plasma glucagon increased transiently after injection of the insulins. The pharmacokinetics and pharmacologic actions of human insulin were essentially similar to those of porcine insulin. The area under the time-concentration curve (AUC) of IRI was slightly greater after s.c. injection of human insulin than after that of porcine insulin at the 0.05-U/kg dose level. Because no significant difference was observed in the plasma C-peptide level and no such difference was noticed in the AUC of IRI between the two insulins after i.v. injection, the bioavailability of human insulin seemed to be greater than that of porcine insulin after s.c. injection. The plasma glucose-reducing effect was slightly greater after s.c. injection of 0.05 U/kg of human insulin than after s.c. injection of the same dose of porcine insulin. Apart from symptoms of hypoglycemia (sweating, palpitations, and hot flushes), no other unwanted reactions were observed after administration of either insulin. These results suggest that human insulin has a usefulness similar to that of porcine insulin in clinical practice. The clinical relevance of the slight difference in bioavailability observed in the s.c. study remains to be investigated.
Complaints after photorefractive keratectomy (PRK) include both severe pain and a decrease in quality of vision due to corneal subepithelial haze. We suspect that one of the causes of these troubles is a temperature rise at the corneal surface during ablation. We cooled down the eye with cold balanced salt solution before and after photorefractive keratectomy. Postoperatively, this reduced severe pain, subepithelial corneal haze and any damage to the corneal endothelium.
Separation of acrosome-reacted viable sperm is an important problem in the analysis of sperm function and the micromanipulation of gametes such as the microinjection of sperm into ooplasm and subzonal sperm insertion. In the present study, we collected acrosome-reacted sperm selectively using paramagnetic immunobeads, referred to as MH61-beads, which attach themselves to the head of the acrosome-reacted sperm and are recovered by a magnet. The maximum number of sperm was recovered by the MH61-beads when 3 x 10(5) sperm/100 microliters and 6 x 10(5) MH61-beads were coincubated for 1 hour. The recovery of sperm was significantly correlated with the percentage of the acrosome reaction in the capacitation medium. After incubation for 12 hours, 2.4-9.0 x 10(4) sperm could be collected, and the percentage of viable and acrosome-reacted sperm was 28-60%. To improve the recovery of the acrosome-reacted sperm, we utilized follicular fluid, which induces the acrosome reaction. Addition of follicular fluid to the capacitation medium at a final concentration of 20% induced the acrosome reaction and increased the recovery of acrosome-reacted sperm significantly. In conclusion, separation of acrosome-reacted sperm by the MH61-bead-binding method could become a useful technique in research of sperm function and gamete micromanipulation.