Insulin-stimulating protein from human plasma.

An insulin-stimulating protein was isolated from human plasma by a procedure involving Sephadex G-100 chromatography and reverse-phase HPLC. The isolated material gave a single band on SDS-polyacrylamide gel electrophoresis. This protein itself had no insulin-like activity, but enhanced fatty acid synthesis from glucose in rat adipose tissue explants in the presence of suboptimal concentrations of insulin. It also stimulated the effect of insulin on CO2 liberation from glucose by isolated rat adipocytes and increased the maximal response to insulin.

Contained rupture of the aneurysm of common iliac artery associated with pyogenic vertebral spondylitis.

STUDY DESIGN: A case report. OBJECTIVES: To report and discuss a case of contained rupture of the aneurysm of common iliac artery associated with pyogenic vertebral spondylitis, so that investigators and practitioners may avoid the diagnostic and therapeutic pitfalls associated with pyogenic vertebral spondylitis and aortic disease. SUMMARY OF BACKGROUND DATA: Pyogenic vertebral spondylitis is a rare disorder that may have serious consequences, including death, if it is not diagnosed promptly and treated effectively. The association of pyogenic vertebral spondylitis with infection of the aorta is a rare but potentially fatal condition that requires prompt diagnosis and aggressive surgical and medical therapy. To our knowledge, this is the first report of a contained rupture of the aneurysm of common iliac artery case associated with pyogenic vertebral spondylitis resulting from an infection with Bacteroides fragilis,although Salmonellae infections are commonly associated with vertebral osteomyelitis and lesions of the contiguous aorta. METHODS: A 60-year-old man with chronic lower back pain began to experience a severe pain and had increased difficulty in walking. An MRI scan showed an increased signal in the L4-L5 disc space and an abscess extending into the spinal canal. The presumptive diagnosis was infective spondylitis. While performing a CT-guided needle biopsy, an unexpected contained rupture of the aneurysm of common iliac artery was discovered. RESULTS: A wide resection of all infected tissue, including the right common iliac artery and bony lesions, was performed in combination with antimicrobial therapy. A cryopreserved aortic allograft was used to reconstruct the artery, and an iliac strut graft was used to fill the debrided vertebral cavity. The patient's postoperative recovery was uneventful. CONCLUSION: The coexistence of pyogenic vertebral spondylitis and lesions of the aorta is rare, but may be lethal if not diagnosed promptly and treated effectively. Even if a patient's condition is stable and the hematocrit is normal, it is important to consider the possibility of a contained rupture of a mycotic abdominal aneurysm in all patients with vertebral osteomyelitis who have acute episodes of unusual severe back pain. CT is sometimes more beneficial than MRI in the identification and characterization of contained rupture of aneurysms.

Plasminogen activator activity in the bovine oocyte-cumulus complex and early embryo.

In this study fibrinolytic assay systems were used to assess the plasminogen activator (PA) potential and plasmin generating ability of oocyte-cumulus complexes isolated from preovulatory bovine follicles (2-8 mm diameter) and of fertilized oocytes from the day of fertilization up to, and including, the hatched blastocyst stage (day 12). During embryo development, the culture medium was changed every 24 hr and samples examined for PA activity. Irrespective of the stage of maturity, no plasminogen or PA could be detected in unfertilized oocytes from which the cumulus layer had been removed. Both plasminogen and PA were found in the cumulus layer indicating that this, rather than the oocyte, was the source of these proteins in the oocyte-cumulus complex. Following oocyte fertilization, no PA activity was detected in either the developing embryo or in the culture medium before day 7. When the embryos had developed to the expanded blastocyst stage, days 7-8, PA production began with activity being detected in both the embryos and their culture medium. Between days 8 and 12, when embryos had reached the hatched blastocyst stage, the PA activity had increased significantly (p<0.05). Analysis of the culture media confirmed this increase in production of PA activity and, based on zymography, it was estimated that the molecular weight of the PA was 78 k daltons.

Bovine erythrocyte haemolysates enhance plasminogen activation by tissue-type plasminogen activator.

An active fraction that accelerates plasminogen activation by tissue-type plasminogen activator (t-PA) was purified from a haemolysate of bovine erythrocytes. When the haemolysate was mixed with t-PA, it produced a 2- to 3-fold increase in plasminogen activation as measured by an insoluble fibrinolytic assay system and a soluble amidolytic assay system with the chromogenic substrate S-2251. Zymographic analysis showed that, while the haemolysate increased t-PA activity, it did not alter the electrophoretic characteristics of the t-PA nor did it induce any fibrinolysis in the absence of t-PA or plasminogen. The haemolysate was devoid of plasmin and plasminogen activator activity but was most effective in accelerating plasminogen activation by t-PA in the presence of substrate. Based on the purification characteristics of the active fraction in the haemolysate, it appears to have a molecular weight of less than 10 kDa.

[Management of cesarean section under replacement therapy with factor VIII concentrates in a pregnant case with congenital combined deficiency of factor V and factor VIII].

The congenital combined deficiency of Factor V and Factor VIII, a rare bleeding disorder, was identified in a 25-year-old woman. She was admitted to our hospital with a complaint of genital bleeding. Her prothrombin time and activated partial thromboplastin time were prolonged. She had low levels of Factor V coagulant activity (F. V:C) 14%, and Factor VIII coagulant activity (F. VIII:C), 12%, and normal levels of von Willebrand factor antigen (vWF:Ag), ristocetin cofactor (Rcof) and Protein C antigen. Her Protein C inhibitor level was slightly low. Her Rcof, vWF:Ag and F. VIII:C were elevated following administration of 1-deamino-8-D-arginine-vasopressin (DDAVP), but her F. V:C remained unchanged. Four years later, her F. VIII:C rose to 70% during the course of her pregnancy, but her F. V:C value remained low. It was expected that the vaginal delivery would be possible at the termination of pregnancy. Premature rupture of the membranes and an anomaly of rotation appeared in the course of delivery, however, and cesarean section was accomplished without excess bleeding under replacement therapy with Factor VIII concentrates. These findings suggested that DDAVP and Factor VIII concentrates were useful for management of her delivery. However the mechanisms of the rise of plasma F. VIII:C during pregnancy in a case with congenital combined deficiency of Factor V and Factor VIII are unclear.

[B-chronic lymphocytic leukemia/prolymphocytic leukemia (CLL/PL)--a case report].

A 48-year-old male was admitted to our hospital on April 20, 1989 because of general fatigue and abdominal fullness. Physical examination showed hepatomegaly, massive splenomegaly, and systemic lymphadenopathy. Hematological findings revealed WBC 73,000/microliters, RBC 289 x 10(4)/microliters, Hb 8.0g/dl, and platelet 9.1 x 10(4)/microliters. WBC differential count demonstrated a mixture of 63% matured small lymphocytes and 32% prolymphocytoid cells. Bone marrow aspiration was unsuccessful with a dry tap. Surface marker analysis of peripheral blood lymphoid cells disclosed that they were positive for anti-HLA-DR, CD 5, CD 19, CD 20, CD 21, CD 25, Sm-IgM, Sm-IgD, and Sm-K. He was diagnosed as B-CLL/PL, and treated with VEPA with partial remission. CLL/PL which was advocated by Melo in 1986 is regarded as a distinct clinical entity intermediate between CLL and PLL in clinical and laboratory features. Our case is interesting with regard to good response to combination chemotherapy, though most cases of CLL/PL have a resistance to standard chemotherapy.

[Combination chemo-endocrine therapy of metastatic and stage IV breast cancer with cyclophosphamide, adriamycin, prednisolone and tamoxifen (CAPT)--with special reference to management of brain and liver metastasis].

Sixty-three evaluable patients with metastatic and stage IV breast cancer who had not previously undergone chemo-endocrine therapy were treated with a combination chemoendocrine therapy regimen consisting of cyclophosphamide 100 mg p.o. every day, adriamycin 10 mg i.v. on day 1 to 5, prednisolone 10 mg or 20 mg (20 mg was given on day 1 to 5) p.o. every day, and tamoxifen 20 mg p.o. every day. Adriamycin on day 1 to 5 was repeated three times every two weeks. After a total dose of 150 mg of adriamycin, the patients were changed to maintenance therapy consisting of cyclophosphamide 100 mg p.o., prednisolone 10 mg p.o. and tamoxifen 20 mg p.o. every day. After 72 months of the treatment there were 61 patients good for evaluation, 13 patients achieved a complete response (21.3%) with a median survival of 30.5 months and 18 patients had a partial response (29.5%) with a median survival of 21.0 months, and 30 patients failed to respond (49.2%) with a median survival of 8.5 months. There was a significant difference in survival time between responders (CR + PR) and non-responders (NC + PD) (p less than 0.001). Responses by site were seen in lung 10/18 (55.6%), liver 3/6 (50.0%), brain 2/4 (50.0%), bone 6/17 (35.3%) and soft tissue 14/24 (56.3%). A Satisfactory response for brain and liver metastasis, which are usually viewed as a sign of grim prognosis, was obtained similar to other sites of metastasis. Retreatment with CAPT, which was attempted in patients with secondary brain metastasis who responded to CAPT for initial brain metastasis, was uniformly effective. High ration of androgen to corticosteroid, positive estrogen receptors, long disease-free survival (over two years), premenopausal, high Broca' index (above 110) resulted from the chemo-endocrine therapy regimen CAPT. Toxicity was minimal and consisted of nausea, vomiting, alopecia and leucopenia.

Size of the tumor and other factors influencing prognosis of carcinoma of the head of the pancreas.

Extent of the tumor in carcinoma of the pancreas seems an important factor in stage classification. The purpose of this report is to present the results of studies on the relationship between the size of the tumor (T) and prognosis, and the correlation between the size and the other factors influencing prognosis such as invasion to the pancreatic capsule, invasion to the retroperitoneal tissue, metastasis to the regional lymph node, and so on. T was divided into four groups: T1, T2, T3, T4. Tumor diameter of 2 cm or less was designated as T1, 2.1-4 cm as T2, 4.1-6 cm as T3, and 6.1 cm or more as T4. Case reports of 163 patients who underwent resection of carcinoma of the pancreatic head were submitted by 10 major institutions and retrospectively analyzed. There was significant differences in 1-year cumulative survival rates between T1 and T3, T1 and T4, and T1 and T2 + T3 + T4, but no significant difference in 3- or 5-year survival rates among the T groups. The incidence of no capsular invasion, no retroperitoneal invasion, and no lymph node metastasis had a tendency to decrease along with increase in tumor size and the incidence of definite capsular invasion, and lymph node metastases increase with increase in the size. Attempts to diagnose tumors of less than 2 cm should be made to ameliorate the poor surgical results. However, it should be stressed that even in the T1 group regional lymph node metastases were seen in about half of the cases.

Studies on the biological activity of an insulin-stimulating peptide from a tryptic digest of bovine serum albumin.

A two-chain polypeptide, which corresponds to amino acid residues 115-143 and 144-184(185) of bovine serum albumin, connected to each other by a disulfide bridge, potentiated the effects of insulin on glucose transport and glucose metabolism in isolated rat adipocytes. Although the peptide alone had little activity, it shifted the concentration-response curves of insulin-stimulated D-[1-14C]glucose oxidation, 2-deoxyglucose transport, and lipid synthesis from D-[U-14C]glucose to lower insulin concentrations. It also increased the maximal responses of these parameters to insulin. However, it did not affect insulin binding to adipocytes. The peptide protected insulin considerably from degradation, but this effect alone cannot account for its effect in increasing the maximal responses to the hormone, and even when degradation of a submaximal concentration of insulin was suppressed by bacitracin, the peptide still had an enhancing effect. These results suggest not only that the peptide influences a step distal to receptor-mediated insulin binding but also that inhibition of insulin degradation alone cannot explain its total effect. The peptide lost its insulin-stimulating activity completely when it was further digested with V8 or lysine-specific endopeptidase, or when it was reduced and then carboxamidomethylated or oxidized with performic acid. Similar active tryptic fragments were obtained from human and rat albumins. Insulin-stimulating peptides should be useful in studies on the mechanisms of insulin action including both the sensitivities and responsiveness of target cells to the hormone.

Insulin-stimulating peptide from tryptic digest of bovine serum albumin.

Insulin-stimulating peptide was isolated from a tryptic digest of bovine serum albumin by gel permeation, SP Sephadex column chromatography, reversed phase HPLC and cation-exchange HPLC. This peptide, with a molecular weight of about 8,400, had no insulin-like activity by itself, but enhanced fatty acid synthesis from glucose in rat adipose tissue explants in the presence of suboptimal concentrations of insulin. It also stimulated the effect of insulin on CO2 production from glucose in rat adipocytes, without affecting insulin binding. These stimulations were dose-dependent and were observed at concentrations of more than 2 X 10(-7) M peptide only in the presence of a suboptimal concentration of insulin.

Insulin-stimulating protein from human plasma. Chemical characteristics and biological activity.

A protein that potentiates the action of insulin in vitro was purified from human plasma. When reduced with 2-mercaptoethanol and then carboxymethylated, it yielded a single subunit, indicating that it was composed of two identical subunits connected by a single disulfide bond. This modified subunit tended to inhibit rather than stimulate insulin activity. A distinctive feature of the amino acid composition of this protein (H-ISP) was the absence of histidine, arginine, and tryptophan. The molecular mass, subunit composition, the characteristic amino acid composition and the N-terminal amino acid residue of H-ISP are very similar to those of human plasma apolipoprotein A-II (apo A-II). The isoelectric point of H-ISP was estimated to be 4.91, which is identical with that of the major apo A-II isoform. H-ISP did not itself have insulin-like activity in increasing CO2 liberation from labeled glucose and 2-deoxyglucose uptake by isolated rat adipocytes, but it potentiated the action of insulin in these parameters. It had no appreciable affect on the binding or degradation of 125I-labeled insulin by adipocytes. Like H-ISP, apo A-II isolated from human plasma also had no insulin-like activity by itself, but stimulated the effect of insulin on CO2 production from labeled glucose in isolated rat adipocytes. From these results, it is concluded that H-ISP is identical with the major apo A-II isoform. Incubation of isolated adipocytes with H-ISP resulted in marked increase in the activity of pyruvate dehydrogenase in a dose-dependent manner in the absence of added insulin. H-ISP also stimulated pyruvate dehydrogenase activity in a subcellular system consisting of plasma membranes and mitochondria from rat adipocytes. The effect of H-ISP on pyruvate dehydrogenase activity could be produced by treatment of the isolated mitochondrial fraction alone.

[A neuron-specific enolase (NSE) positive leiomyosarcoma].

A 40-year-old woman with multiple skin tumors was admitted to our hospital on September 5, 1989. Extensive studies by means of light and electron-microscopic examination and immunohistochemical analysis revealed a neuron-specific enolase (NSE) positive leiomyosarcoma. The level of her elevated serum NSE decreased after receiving her initial chemotherapy, but after the fourth session of chemotherapy, the NSE began to elevate again and she died on December 25, 1989. NSE appears to be a useful marker for determining if the neuronal and neuroendocrine cells are normal. This rare case, however, seems to demonstrate that abnormal, nonneuronal cells like a sarcoma also open metabolic pathways to synthesize NSE.