Infection of human natural killer (NK) cells with replication-defective human T cell leukemia virus type I provirus. Increased proliferative capacity and prolonged survival of functionally competent NK cells. Human T-cell leukemia virus type I (HTLV-I) can infect a variety of human cell types, but only T lymphocytes are efficiently immortalized after HTLV-I infection. This study reports an attempt to infect and to immortalize NK cells with HTLV-I. Co-cultivation of freshly isolated NK cells with a HTLV-I-producing T cell line did not result in NK cell infection. However, NK cells activated with an anti-CD16 mAb and co-cultivated with a HTLV-I-producing T cell line were reproducibly infected by HTLV-I. HTLV-I infection was documented in NK cell lines and clones by the detection of defective integrated provirus by both Southern blot and polymerase chain reaction analysis. Although HTLV-I-infected NK cells produced viral proteins, they did not produce infectious viral particles. HTLV-I-infected NK cells were phenotypically indistinguishable from their uninfected counterparts (CD16+, CD2+, CD56+, CD3-). They also retained the ability to mediate both natural and antibody-dependent cell cytotoxicity. The IL-2-dependent proliferation of HTLV-I-infected NK cells was significantly greater than that of uninfected NK cells. The doubling time of this infected population was reduced from 9 days to 3 days, and the overall survival of the culture in the absence of restimulation was extended from 5 wk to 18 wk. Unlike T lymphocytes, HTLV-I-infected NK cells were not immortal, implying a fundamental difference between these two lymphocyte populations.


Generation and characterization of cloned T helper cell lines for anti-DNA responses in NZB.H-2bm12 mice. We have previously demonstrated that the introduction of the bm12 mutation into NZB mice results in animals that spontaneously produce high titer IgG autoantibodies to dsDNA. The observation that NZB.H-2bm12 develop lupus although NZB.H-2b control mice do not, provides a unique system to study the role of Th cells in the production of antibodies to dsDNA. We have isolated, in the absence of a known stimulating autoantigen, a series of seven autoreactive T cell clones that provide help in vitro for the production of IgG anti-dsDNA antibodies by syngeneic B cells. The data on these seven cloned T cell lines was compared to two cloned T cell lines specific for keyhole limpet hemocyanin. The seven cloned T cell lines, coined clones 19D, 23G, 410F, 410H, C1, C15, and C52 all show significant help in vitro for production of IgM and IgG antibodies to ssDNA and dsDNA; antibody levels increased 7- to 30-fold compared to cultures without T cells. Clones C1, C15, and C52 were furthered studied and were shown to provide help for IgM antihistone and anti-OVA responses but provided significantly less help for IgG antibodies. In contrast, keyhole limpet hemocyanin-specific cloned T cell lines TK2 and TK5 provided help for IgM antibodies to ssDNA, dsDNA, and histone, but failed to significantly increase IgG antibodies to ssDNA, dsDNA, or histone. The cloned T cell lines were restricted to H-2bm12 and proliferated only in response to APC from NZB.H-2bm12 and B6.C-H-2bm12 but not NZB.H-2b or NZB.H-2d mice; their in vitro helper activity was inhibited by antibodies to class II. All cloned T cell lines expressed Thy-1, CD5, and TCR-alpha/beta. Three of the seven clones used TCR-V beta 4. However, the V beta expression of the four remaining autoreactive T cell clones could not be determined. All of the autoreactive cloned T cell lines produce significant IL-4 but no detectable IL-2 or IFN-gamma. We believe that HPLC-purified peptides eluted from I-Abm12 molecules from APC can potentially provide insight on the putative autoantigen.


Medical admissions: a confusing disease. It is strictly for admission to undergraduate medical courses. (i) 50% of seats should be reserved for local candidate on merit of HSC, with academic year starting after the HSC results. (ii) 20% of seats should be reserved for candidates coming from rural areas, on the basis of merit at HSC, with academic year starting after HSC results. (iii) 30% of seats to be filled by an all-India competitive examination. The academic year to start 6 months after the HSC results. A ceiling of minimum number of marks and a minimum number of attempts for eligibility to the all-India PMT would make the conduct of such an examination more practical.


Primary malignant lymphoma of the gastro-intestinal tract: a clinicopathological study of 24 cases. Twenty-four cases of primary lymphoma of the gastro-intestinal tract were diagnosed during the period 1970 to 1991. There was a preponderance of males and the male to female ratio being 1.4:1. Age ranged from 9-70 years, mean 32.2 years. Small intestine was involved in 50% cases, large bowel in 9 cases (37.5%) and stomach in 3 cases (12.5%). There were 5 cases (20.8%) of Hodgkin's disease and 19 cases (79.2%) were of non-Hodgkin's lymphoma. All cases of gastric lymphoma complained of epigastric pain, weight loss and vomiting. In lymphoma of small intestine, 8 patients complained of pain associated with vomiting and 6 patients complained of distension of abdomen. In large bowel lymphoma, pain in right iliac fossa was complained by 4 patients and bleeding per rectum by 3 patients. Out of all the 24 cases, changes in bowel habit were noted in 15 patients and occult blood was positive in 13 cases. Palpable abdominal mass was noted in 14 patients. Histomorphologically, all the 3 cases in the stomach were of lymphocytic lymphoma diffuse type. Out of 19 non-Hodgkin's lymphoma, 15 were of lymphocytic lymphoma and 4 were of histiocytic lymphoma.


Iatrogenic dry eye: late effect of topical steroid formulations. A total of 69 eyes of 35 patients receiving local corticosteroid therapy over a period of 3 months to 2 years for different eye ailments were studied. It was observed that there was a significant reduction (p < 0.05) of Schirmer's test I, as well as tear film break up time values with an increase in the duration of treatment. Of the various preservatives used with the various brands of steroids it was observed that benzalkonium chloride is most commonly associated with dry eye syndrome. The complications of prolonged topical steroid therapy included steroid induced glaucoma (4 eyes), lenticular opacities (4 eyes), epitheliopathy (2 eyes) and flare up of silent viral keratitis (one eye).


Comparative study of midtrimester termination of pregnancy using hypertonic saline, ethacridine lactate, prostaglandin analogue and iodine-saline. The study consisted of terminations of 200 cases of second trimester pregnancies ranging from 14 weeks to 20 weeks. Out of these 200 cases, in 50 cases intra-amniotic instillation of 20% hypertonic saline (200 ml) was done after withdrawing 35-200 ml of amniotic fluid. Ethacridine lactate was instilled in 50 cases extra-amniotically. Prostaglandin F2 alpha was injected intramuscularly at regular intervals in 50 cases. Fifty cases of pregnancies were terminated with extra-amniotic instillation of 5% povidone-iodine solution mixed with normal saline. Comparison was made among all the methods regarding instillation-abortion interval, completeness of abortion, failure of the procedure and postoperative complications. Solution of 5% povidone-iodine in normal saline was found to be comparable in all aspects to other methods and above all a much cheaper alternative for poor patients. Success rate was highest with iodine-saline solution (100%) followed by ethacridine lactate (98%), hypertonic saline (96%) and lowest with prostaglandin F2 alpha (90%).


The blood group and haemoglobin types of the Santals. A total of 488 blood samples from tribal as well as non-tribal donors of Bihar and West Bengal were analysed for presence of abnormal haemoglobin variants. Out of 488 samples ABO and Rh blood groups were studied in 468 samples. Predominance of group B was noted in both the tribal and non-tribal groups and absence of Rh(d) was noted in non-tribal population with the exception of the tribals where 4.42% Rh(d) gene was found. Incidence of haemoglobin E gene was very low in most cases but relatively higher in non-tribal populations, while the range of raised haemoglobin A2 varied from 5.84% to 8.80%.


The natural history of smoking.

The natural history of smoking. This study is a small collection of the natural history of smoking of 865 persons - all smokers and majority (97.11%) of them were males, more than 80% were between 21-50 years, most (90%) were Hindus and 75% were service holders. It revealed that most of the subjects started smoking between 10 and 25 years of age, frequently being requested by their friends but felt nothing mentionable at their first experience. Most of them (64%) were smoking for more than 10 years, 48.56% were smoking more than 10 cigarettes in a day, filter-tipped more often (by 64% subjects) and without any history of break by 56% subjects. Majority (80%) used to inhale the smoke and 40% had no specific time of choice. Most of the subjects (80%) like to smoke in deep thoughts, 66% during excitement, 60% when depressed, 82% when relaxed and 62.5% when alone. But most of them (64%) did not smoke when busy. Majority (70.87%) felt relaxed when smoking. Most of the people (45%) stood reasonable having no cigarette in their stock. Majority (84%) used to smoke even when not offered. Tobacco was consumed largely as smoke by 75% subjects; 78% subjects smoked only cigarettes and about half of the smokers smoked even when they did not enjoy it.


Angiomyolipoma of kidney.

Angiomyolipoma of kidney. Angiomyolipoma is an uncommon benign tumour accounting for less than 1% of all surgically excised tumours of the kidney. It may present as an isolated lesion or in association with tuberous sclerosis. An impalpable tumour is often detected incidentally during investigation of a case for some urinary tract pathology. CT scan has proved to be a useful investigation for pre-operative diagnosis; it may however, be of no value if the fat content of the tumour is low. Here a case of angiomyolipoma of left kidney is reported where pre-operative diagnosis was suggested by CT scan and histopathology confirmed it.


Rare variant of acute promyelocytic leukaemia. Acute promyelocytic leukaemia (APL) is a peculiar sub-type of acute myeloblastic leukaemia characterised by presence of atypical promyelocytes in bone marrow and peripheral blood and common occurrence of haemorrhagic episodes associated with disseminated intravascular coagulation. Two morphological forms of APL are recognised--typical hypergranular and microgranular or M3 variant. This microgranular form of APL is rare but has got some peculiar features and often can be diagnosed by peripheral blood smear examination alone without bone marrow examination. Three cases of microgranular form of APL observed during a period of 6 months are reported here.


Serum erythropoietin titers in hematological malignancies and related diseases. Serum erythropoietin (Epo) titers in patients with various hematological malignancies and related diseases were determined by radioimmunoassay. Serum Epo titer was inversely correlated with hemoglobin concentration in iron deficiency anemia, aplastic anemia, myelodysplastic syndromes (MDS), acute leukemia, malignant lymphoma, multiple myeloma and myelofibrosis, but there was no correlation between serum Epo titer and hemoglobin concentration in chronic myelogenous leukemia or polycythemias. Serum Epo titers in aplastic anemia were much higher than those in iron deficiency anemia. Serum Epo titers in MDS, malignant lymphoma and multiple myeloma differed considerably among patients. Serum Epo titers in untreated polycythemia vera were significantly lower than in treated polycythemia vera or secondary polycythemia.


Peripheral blood hematopoietic progenitor cells in beta-thalassemia major. Colony assays in methylcellulose (semisolid medium) of erythroid precursors and granulocyte-macrophage precursors from the peripheral blood of 34 patients with beta-thalassemia major and 31 age-matched normal controls were studied. In patients homozygous for beta-thalassemia, a significant increase of circulating erythroid progenitor cells to 3-5 times of normal controls was observed. Furthermore, the number of circulating colony forming units-granulocyte-macrophage (CFU-GM) and colony forming units-mixed (CFU-Mixed) also increased significantly. In these subjects, there was a linear relationship between the number of burst forming units-erythroid (BFU-E) and CFU-GM. The serum level of erythropoietin (EPO) was also significantly higher than that of normal controls (p < 0.05). The increased hematopoiesis, indicated by an increased number of circulating hematopoietic progenitor cells and an elevated serum level of EPO, suggests that the physiologic regulation of erythropoiesis still operated in patients with beta-thalassemia major.


An in vitro thymidine incorporation assay for human cancers: technical details revisited. Among the choices of laboratory methods for in vitro testing of human tumor chemosensitivity, a thymidine incorporation assay (TIA) has been described. Advantages of a TIA over a colony-forming assay include greater probability of tumor growth, requirement for fewer tumor cells, less dependence on an absolute single-cell suspension, shorter time to completion, and the ability to measure up to three logs cell-kill. Specific technical details which are reported in the literature for performing a TIA are incomplete and/or conflicting between one laboratory and another. This paper reports details of methods designed to remove unbound 3[H]-thymidine from the agarose, standardize background counts, and determine optimal cell-plating densities, labeling time, optimal concentration of 3[H]-thymidine, and the best day after seeding cultures to add the 3[H]-thymidine. Care has been taken to compare the end point of drug sensitivity determinations following these methodologic changes, both among serial TIAs as well as with colony-forming assays.


Cloning of human tumor cell lines in porous glass capillary tubes: a further development of the human tumor stem cell assay. The conventional human tumor stem cell assay for cloning tumor cells for drug sensitivity testing is limited by its inability to test drug combinations. In an attempt to overcome this limitation, we cloned tumor cell lines within porous glass capillary tubes. In contrast to plastic porous tubes, the porous glass membranes were transparent, and colony formation could be judged on an inverted microscope. Human as well as animal cell lines showed sufficient colony growth. Colonies formed within these porous tubes were homogeneously distributed, and their morphology was similar to those formed in the common stem cell assay. Cloning efficiency and colony size depended on the mean pore diameter of the glass membrane, with best colony growth within tubes with a pore diameter ranging from 8.5 nm to 14 nm. A linear relationship between number of cells seeded and number of grown colonies could be demonstrated for the cell lines MDA-231 and Colo 201. Colony growth achieved within porous glass capillary tubes is comparable to that achieved in Petri dishes and in nonporous tubes. We conclude that the porous capillary cloning system meets the basic suppositions for a quantitative cloning assay. Moreover, the porosity of the glass membrane offers the possibility of variable perfusion of medium and drugs. Further investigations will focus on various perfusion modalities and chemosensitivity testing.


Influence of feeding regimen and postnatal developmental stages on antibacterial activity of pancreatic juice. Antibacterial activity of pancreatic juice in the pig (n = 8) was investigated during early postnatal development and in cattle (n = 6) receiving a different feeding regimen. For pancreatic juice collection, a catheter was surgically implanted in the pancreatic duct. Reintroduction of pancreatic juice was achieved through a T-shaped cannula in the duodenum. Pancreatic juice was collected for 30 min in all cases. In piglets, collections were carried out at 2, 5-6, and 7-10 wk of age, and in cattle, after a standard meal, 48 h starvation, and following 24 h intraduodenal glucose infusion. Antibacterial activity was tested on Micrococcus Pyogenes strain ATTC 6538P by disc agar diffusion technique using nonactivated pancreatic juice, before and after heat treatment for 15 min at 65 and 100 degrees C, respectively. Piglets showed a significant rise in antibacterial activity of pancreatic juice after weaning. In cattle, 48 h of starvation resulted in a marked suppression of antibacterial activity. This activity was found to be normal after a standard meal and comparable to that after 24-h intraduodenal glucose infusion. Heating of pancreatic juice to 65 degrees C caused a 35% increase in the antibacterial potency, whereas heating to 100 degrees C completely abolished it. Additionally, dilution of pancreatic juice to 1:10 did not affect antibacterial potency.


Fibrinolytic enzymes in ascites during experimental acute pancreatitis in rats. The ascites accumulating during acute pancreatitis contain proteases that play a role in the progression of this disease. The proteases of the fibrinolytic system in the ascites were therefore studied in experimental acute pancreatitis induced in rats. Synthetic substrate assay and the fibrin plate method revealed high activities of proteases, including plasminogen activator, in the ascites. The plasminogen activator had a mol wt of about 50,000 by zymography. The plasminogen activator adsorbed on Lys-sepharose from the ascites was observed at the 100,000 mol wt position and in the 50,000-100,000 mol wt range on zymography and appeared at the 50,000 mol wt position after treatment by concentration. Its activity was enhanced by trypsin treatment. In other experiments, when incubated homogenate of normal pancreas lacking in zymographic activity was injected intraperitoneally into healthy rats, the recovered fluid displayed lytic zones between the 100,00 and 50,000 mol wt positions. These findings suggest that the ascites contained plasminogen activator, part of which was released by intrapancreatic substances and was present in the precursor form.


Effects of gastric fundectomy and antrectomy on the exocrine pancreas in the hamster. The effect of gastric fundectomy and antrectomy on growth of the exocrine pancreas was studied in hamsters over 5 and 25 d. Sham-operated animals served as controls. After 5 d, basal plasma gastrin concentrations were significantly increased in fundectomized animals (80.3 +/- 20.6 pmol/L) and significantly decreased in antrectomized animals (11.6 +/- 1.1 pmol/L) as compared with the controls (20.0 +/- 1.7 pmol/L). Similar differences were present among the 25-d groups, whereas basal plasma cholecystokinin (CCK) concentrations did not differ significantly between any groups at any time. At 5 d after fundectomy, there was a significant increase in pancreatic tissue [3H]-thymidine uptake and total DNA content, both of which were reduced 5 d after antrectomy. Autoradiography showed significantly increased [3H]-thymidine labeling index of acinar, intralobular duct, and centroacinar cells of the pancreas at 5 d after fundectomy. The increased intralobular duct cell labeling index persisted 25 d after fundectomy. Labeling indexes after antrectomy did not differ significantly from those in the controls, although antrectomized animals had the lowest values in all three cell compartments at 25 d. At 25 d, pancreatic wet wt and total DNA and protein content were significantly increased after fundectomy and significantly reduced after antrectomy. These findings indicate that fundectomy in the hamster induces pancreatic exocrine tissue hyperplasia and hypertrophy, whereas antrectomy leads to retardation of pancreatic growth.


Ductal adenocarcinoma of the pancreas. Histopathological features and prognosis. Between 1972 and 1987, curative surgical resection (RO) was performed in 81 patients with ductal adenocarcinoma of the pancreas. In this study, slides from surgical specimens were reviewed, and histopathological features of the carcinomas were retrospectively reevaluated. Tumor stage was the most important prognostic factor: In UICC stages I, II, and III, the median survival times were 13, 16, and 8 mo, respectively. Lymph node involvement and direct extension of the tumor into adjacent peripancreatic tissue, as well as invasion into peripancreatic organs were found to significantly influence survival. Tumor infiltration of the lymphatic vessels was present in 74% of the resected carcinomas and significantly correlated with survival time. There was no relationship between survival and tumor size; furthermore, histological grade of differentiation, age, and sex had no influence on prognosis.


Effects of intraluminal peptide YY on pancreatic function. Peptide YY (PYY) is released postprandially into both the circulation and the distal intestinal lumen. While circulating PYY inhibits pancreatic secretion and insulin release, the effects of intraluminal PYY on pancreatic function are unknown. The aim of the present study was to evaluate the effect of exogenous, intraileal luminal PYY on pancreatic exocrine function and fasting glucose levels. Chronic pancreatic and ileal fistulae (50 cm from the ileocecal valve) were created in nine mongrel dogs. The animals were given intravenous infusions of secretin (125 ng/kg/h) and cholecystokinin octapeptide (CCK-8, 50 ng/kg/h) for four hours. At the beginning of the second hour, either normal saline or PYY, at low [physiologic (2 ng/min)] or high [supraphysiologic (50 ng/min)] levels, was infused antegrade into the ileal fistula for two hours. Pancreatic juice was collected for PYY and glucose levels. Ileal luminal PYY infusions had no effect on pancreatic bicarbonate or protein output. Fasting serum PYY and glucose concentrations were unaffected by either dose of intraluminal PYY. We conclude that ileal luminal PYY does not influence pancreatic exocrine function or fasting glucose levels.


Influence of a platelet-activating factor antagonist on severe pancreatitis in two experimental models. This study investigates the role of platelet-activating factor (PAF) in experimental pancreatitis. The concentration of PAF quantified in ascites of bile-induced pancreatitis by radioimmunoassay (RIA) ranged from 3.67 +/- 0.39 pmol/mL 2 h to 0.954 +/- 0.39 pmol/mL 10 h after injection of taurocholate. Administration of a potent PAF antagonist, WEB-2170, prior to injection of taurocholate prolonged mean survival time in rats receiving i.v. camostate and albumin (46.4 h, n = 15, vs controls 38.3 h, n = 13). However, the survival rate after 72 h was not improved. The histologically estimated severity of pancreatitis and pancreatic enzymes in blood, tissue, or ascites was not affected. WEB-2170 had no effect on survival when injected simultaneously with taurocholate into the pancreatic duct or given i.v. after induction of pancreatitis (1, 0.1, or 0.01 mg/kg WEB-2170 vs controls). Subcutaneous injection of 10 mg/kg WEB-2170 also did not improve survival in pancreatitis induced by choline-deficient, ethionine-supplemented diet in mice. It is concluded that administration of a PAF antagonist after the onset of severe experimental pancreatitis does not influence its outcome, although activation of PAF may play a role in the pathogenesis of pancreatitis.


Isolation of adult pig islet. In vitro assessment and xenotransplantation. This study was conducted to develop an isolation method for adult pig islets and to investigate in vivo function after xenotransplantation as well as in vitro function of isolated islets. From the splenic portion of the glands (n = 20), 3277 +/- 645 islets per gram of pancreas were isolated and recovered by a five-step dextran discontinuous density gradient method. Purity of the final preparation obtained from 20 different consecutive pig pancreata was 80-90%. In vitro incubation study revealed a significant insulin release from isolated adult pig islets in response to glucose stimulation. In vitro perifusion study demonstrated a biphasic insulin response to glucose stimulation from isolated islets. Xenotransplantation of approx 2000 isolated adult pig islets into the portal vein of diabetic Wistar rats (n = 8) significantly reduced serum glucose levels from 431 +/- 24 mg/dL to 173 +/- 18 mg/dL 24 h after transplantation. This study provides a development of an isolation method for adult pig islets and demonstrates that isolated islets are viable and functioning both in vitro study and in vivo study of xenotransplantation.


Solitary true cyst of the pancreas in an adult. A 35-yr-old woman was presented to the hospital because of nausea 20-30 min after eating, associated with weight loss. A CT scan revealed a cystic lesion in the neck of the pancreas. At operation, a 3-cm diameter cyst was excised from the pancreas. Endoscopic retrograde pancreatography (ERP) was used intraoperatively to identify the transected pancreatic duct, after which a Roux-Y pancreaticojejunostomy was performed. Histologically, the cyst proved to be unilocular and was lined with cuboidal epithelium. The natural history of this lesion is unknown as only four previous case reports of unilocular true cysts in adults have been reported in the English literature. These are reviewed.


Submandibular gland as a site for islet transplantation. Homologous transplantation of islets of Langerhans into the submandibular glands of Syrian hamsters was successful in 8 out of 10 recipients. The technique was simple and led to formation of islets of various sizes within the parenchyma of the gland. The morphology and endocrine cell patterns of this islets were identical to pancreatic islets. The advantage of this model for islet transplantation is discussed.


Detection of deep vein thrombosis with a computerized strain gauge plethysmograph. Forty patients suspected clinically the first episode of deep vein thrombosis (DVT) were studied using a new computerized strain gauge plethysmographic (SGP) system, the Phlebotest. A capacitance mode was used to secure the obtaining of maximum venous filling before deflation in every determination. Following parameters were calculated automatically and instantly: the maximum volume change (V sec), the outflow rate during the first second after deflation (F 1.0), the expelled volume in four seconds (EV 4.0) and the surface area over the curve during the first four seconds after deflation (IND). The eighty lower limbs of the patients were divided into three groups according to the Duplex scanning results: proximal (n = 19), distal (n = 13) and no DVT (n = 48) groups. The diagnostic accuracy of the Phlebotest parameters was determined and compared with that of another widely used SGP system, the Perivein. In this case the maximum volume change and the maximum venous outflow (MVO) were calculated manually. The results of the two systems were compatible. For the detection of proximal DVT, the parameters of venous outflow were better than maximum volume change and the best single parameter was maximum venous outflow: F 1.0 for Phlebotest and MVO for Perivein. The two parameters had a similar sensitivity of 79% when 50 and 27 ml/100 ml/min were used as the lower limits of normal respectively. The best criterion for the diagnosis of proximal DVT was F 1.0 < 50 ml/100 ml/min plus the EV 4.0/V sec ratio < 60%, which had a sensitivity of 90%, specificity of 96% and overall accuracy of 94%.(ABSTRACT TRUNCATED AT 250 WORDS)


Natural history and evolution of peripheral obstructive arterial disease. The natural history of peripheral arterial occlusive disease is discussed. Severe limb-threatening ischemia is the most serious consequence of chronic arterial occlusive disease. Severe ischemia and amputation can be considered as an endpoint in peripheral vascular disease. Severe limb ischemia is relatively uncommon in isolated aortoiliac disease and this is more than twice in patients with either femoropopliteal or multisegmental disease. Subsequent studies have also demonstrated that both smoking and diabetes are associated with a substantial risk for sudden ischemia. A clear majority of about 50% deaths are caused by associated coronary artery disease, 15% to stroke and 10% to vascular disease in the abdomen. Ankle systolic blood pressure is one of the most significant factors in the progression of peripheral arterial occlusive disease and also for cardiovascular mortality. In the future, men need to know how therapies as exercise, during regimens would influence the most frequent complications besides severe limb ischemia, namely brain infarction and coronary artery disease.


The grade of carotid stenosis as a prognostic factor during a 14 years' follow-up in 40 angiographically documented patients with cerebrovascular disease. Forty consecutive patients underwent extracranial arteriography between 1974-1976 because of cerebrovascular disease and were treated nonoperatively. This period was chosen to ensure a sufficiently long follow-up. The treatment selection was made individually by the patient's doctor. Deaths (n = 22) during 14 years of follow-up were caused by cerebrovascular disease in 41%, by coronary heart disease in 27% and other in 32%. The median age of decreased patients (group A) was 60.5 years at the time of arteriography and of patients alive after follow-up (group B) 55.5 years, respectively (p = 0.007). Findings were more severe arteriographically in group A than in group B. Twelve patients had occlusion or > 50% stenosis in the common carotid artery or the internal carotid artery in group A and three in group B (p = 0.014). During follow-up there were 17/22 cerebrovascular events in group A and 8/18 in group B (p = 0.033). Smoking was significantly more common in group A (p = 0.002). When the patients were divided on the basis of haemodynamically significant carotid artery stenosis, patients with significant stenosis (group C) proved to be older than patients with non-significant stenosis (group D), 62.0 versus 57.0 years, respectively (p = 0.001). The incidence of cerebrovascular events was equal in these groups. Our data suggest that patients with over 50% carotid stenosis have a poor long term prognosis, associated with age and smoking.


Is emergency aorto-superior mesenteric artery by-pass worthwhile? Out of 9 superior mesenteric artery (SMA) revascularizations made in emergency, only an early direct aortic reimplantation was successful. The other cases were bypasses and their failures were due either to shock at revascularization (2 cases), or to thrombosis of a by-pass (4 cases), or to persistent ischemic lesions (2 cases). It is suggested: (1) that late mesenteric revascularization should be excluded and let place to extensive bowel resection, (2) that, in emergency, direct SMA aortic reimplantation is preferable to aorto-SMA by-pass.


Correlations among capillaroscopic abnormalities, digital flow and immunologic findings in patients with isolated Raynaud's phenomenon. Can laser Doppler flowmetry help identify a secondary Raynaud phenomenon? Thirty subjects with a diagnosis of isolated Raynaud's phenomenon, according to anamnestic and objective criteria, were cross-evaluated by various methods: laser Doppler flowmetry (LDF) during a standardized cold and rewarming test, nailfold capillaroscopy, immunological and other laboratory parameters, to assess the diagnostic and prognostic significance of each method. Correlations were also assessed among disease duration, capillaroscopic pattern score, quantitative digital flow values and laboratory parameters. Nearly 30% of patients showed immunologic abnormalities (ANA positivity at variable titres, ENA, hypocomplementemia, immunocomplexes); 16% of patients had a pathologic capillaroscopic pattern, not well correlated with immunologic findings; a characteristic cold stop reaction of digital flow (partial or total) was detected in 86% of the subjects; in ANA+ pts., ANA titers were positively correlated with the intensity and length of stop reaction (Trec). A significant correlation between digital flow parameters and the capillaroscopic score was also found in each considered group. Our results outline the relevance of LDF, during a standardized thermic test, to evaluate apparently primary RP, because even if a definite scleroderma-like capillaroscopic pattern is absent, this flowmetric method may detect potentially secondary RP patients.


[Immunological and cytogenetic studies on early spontaneous abortions]. To reveal the role of maternal immunological mechanisms in early spontaneous abortions, the blocking effect (BE) of the maternal serum was evaluated in a mixed lymphocyte culture, and the chromosome constitution of the abortuses was examined. 1) The mean BE in non-pregnant women as a control (n = 7) was 0.86%. 2) The mean BE in women with an uneventful course of pregnancy (n = 21) was 24.1%, which was significantly higher than that in the non-pregnant women (p < 0.01). 3) Seventeen of the 36 abortuses (47.2%) were chromosomally abnormal. 4) The mean BE among all the patients with spontaneous abortions (n = 36) was -9.8%, which was significantly lower than that in the pregnant women (p < 0.01). However, no difference in mean BE was observed between the patients with chromosomally normal (-8.4%) and abnormal abortuses (-11.2%). 5) In addition, no difference in mean BE was observed between the patients with an abortus with a positive fetal heart beat and the patients with an abortus with a negative fetal heart beat. These results indicate that the decreased BE observed in patients with early spontaneous abortion is a consequence of abortion rather than the cause of abortion.


Renal artery stenosis in patients with peripheral vascular disease and its correlation to hypertension. A retrospective study. To estimate the frequency of renal artery stenosis, and to detect the correlation between renal artery stenosis and hypertension, 450 consecutive patients with peripheral vascular disease (PVD) were selected. All subjects had undergone aorto-femoral conventional angiography. For a possible association with renal artery stenosis, risk factors, clinical and angiographical variables were evaluated e.g.: age, sex, diabetes mellitus, smoking habits, use of antihypertensive drugs, serum creatinine, serum cholesterol, ECG pathology, side of the renal artery lesion, bilateral stenoses, post-stenotic dilatation, number of renal arteries, aortic atherosclerosis, size of the kidneys and angiography induced renal dysfunction. Renal artery stenosis (RAS) was found in 49.1%, 117 patients had a moderate and 104 had a severe stenosis. Of the 221 patients with a renal artery lesion, 44 were normotensive, 177 hypertensive. Hypertension was significantly correlated to RAS. An association was also found for age over 70 years, smoking and pathologic ECG. It is concluded that renal artery stenosis is very common in a population with peripheral vascular disease, and the results achieved from this study makes it worthwhile to identify possible functional markers in a prospective study.


Medical treatment of critical limb ischaemia: encouraging results if candidates selected early, therapy based on physiological principles and long-term control established. Medical therapy of CLI is only indicated in patients unsuitable for vascular reconstruction or angioplasty. The approach includes early detection of subjects at risk, observation of physiological principles of therapy and long-term control. The finding of ankle pressure < or = 50 mmHg is of particular value in early detection because of high specificity with respect to CLI. Limb dependency increases skin flow in CLI while in patients with moderate ischaemia vasoconstriction occurs. Positioning of the limb thus contributes to therapeutical effect. Vasodilator substances increase foot blood flow only when administered to patients covered with blankets; improper administration may cause vasoconstriction. When a long-term control programme is established, the number of episodes of CLI decreases as compared to spontaneous course of disease. Rational use of available methods in early diagnosis and therapy considerably improves the prognosis of CLI.


Walk training and drug treatment in patients with peripheral arterial occlusive disease stage II. A review. On the basis of the present studies physical therapy is the most effective basis therapy of peripheral arterial occlusive disease stage II according to Fontaine. The consequent integration of the patients into widespread vascular training groups would be desirable. All present studies with so-called vasoactive drugs led to a statistically significant increase in pain-free walking distance. This is especially true for the substances naftidrofuryl, pentoxifylline, and buflomedil. Nevertheless, these studies do not fully meet the standards set by the GCP or the FDA guidelines. It must also be said that the increase in walking distance by vasoactive substances is less pronounced than the effect obtained by walk training alone. Both the vasoactive therapy and controlled walk training aim at an increase in pain-free walking distance. It is, however, still unclear whether the modes of therapy described influence the primary disease. Angiographically controlled studies are momentarily not available.


[Urinary calcium excretion as an early prediction marker for pregnancy induced hypertension]. Two hundred and forty-nine healthy women at 36 weeks gestation and 38 women with pregnancy induced hypertension (PIH) underwent urinalysis for urinary calcium excretion to obtain the calcium/creatinine ratio. This ratio was significantly lower in the women with PIH than in the healthy pregnant women, and it was especially low in those with PIH who had no family history of hypertension. Three hundred forty-eight healthy women at 20 weeks of gestation underwent urinalysis to determine their calcium/creatinine ratio as a means to predict PIH in patients showing no symptoms. In the primiparous group with no family history 58% of the patients with a low calcium/creatine ratio eventually developed PIH. Using a receiver operator curve, we calculated a predictive threshold calcium value for PIH of 0.06 at six month's pregnancy. Thus urinary calcium excretion may be a useful early marker for PIH.


Preliminary results of a complete study protocol on synthetic vascular graft healing and its complications. The microscopic and anatomic features and bacteriologic culture results of different portions of single, explanted dacron synthetic vascular grafts (SVG) were studied together with patient clinical data. With this complete study protocol a better understanding of the healing process and its associated pathology can be achieved. We studied three, amply distanced graft portions from each of five patients (15 total graft portions) undergoing revision for infectious and non-infectious reasons. We divided the SVG portions studied into a Group 1, with high degrees of graft healing and into a Group 2, with both infection-dependent, early healing complications and perigraft chronic inflammatory reaction-dependent, late healing complications. These late healing complications were found dependent upon a host vs graft reaction. This study confirmed in humans the important role of an internal and external fibrotic graft incorporation in the definitive healing of a SVG. A host vs graft reaction was suggested to be an alternative to the frequently cited low virulent infection pathogenesis of late SVG healing complications. A sure definition and treatment of late SVG healing complications will only be established by means of a complete study protocol performed on a large number of explanted SVGs.


Prevalence of risk factors in patients with peripheral arterial disease. A clinical and epidemiological evaluation. The aim of this study was to evaluate the prevalence of arterial hypertension and other risk factors in patients suffering from peripheral arterial disease (PAD) in two clinical samples (1.: 102 patients with PAD 69 M, 33 F, studied in our angiology laboratory, matched for sex and age with 102 healthy volunteers; 2.: 184 hospitalized patients, 80 M, 104 F, mean age 57.2 +/- 10.8, with PAD) and in two epidemiological cohorts (1.: Trabia Study, 835 subjects; 2.: Casteldaccia Study, 723 subjects). All patients were subjected to a full clinical and laboratory examination, including the determination of the ankle/arm pressure ratio (Winsor index, positive for PAD when lower than 0.95). In the first clinical study we observed a significantly (p < 0.01) greater prevalence of arterial hypertension (51.9 vs 9.8%), hypercholesterolemia (48.2 vs 21.6%), hypertriglyceridemia (53.7 vs 26.1%), smoking habit (64.3 vs 44.2%), and hyperglycemia (26 vs 7,9%) in PAD patients than in controls. In the second clinical study considering separately the patients under and over 65 years, all risk factors resulted to be more prevalent in younger people than in the aged, except for diabetes and hypertension. In our epidemiological experience, the prevalence of PAD increases with aging, above all in males. In the Trabia Study the risk factors, more associated with PAD, were hypercholesterolemia, smoking and obesity (41.18%) in males and hypertension and hypercholesterolemia (33.3%) and obesity (25%) in females. In the Casteldaccia Study the most important risk factors were smoking (64.28%), hypercholesterolemia (42.86%) and hypertriglyceridemia (35.71%) in males, and obesity (60%), hypercholesterolemia (30%) and diabetes (20%) in females. Cholesterol levels and smoking were significantly higher in PAD patients than in the general population, whereas hypertriglyceridemia and glycemia were not. Arterial hypertension was significantly associated with PAD in the Trabia but not in the Casteldaccia Study. Obesity was significantly associated to PAD in females in both studies. In the Casteldaccia Study, lower HDL-cholesterol levels were observed in PAD patients, above all in males, whereas significantly greater Apo-B values and lower Apo-A1 levels (in males) were shown. The different levels of associated risk factors and their prevalence in PAD patients confirm the multifactorial pathogenesis of atherosclerosis. The exact role of each risk factor in the genesis of PAD is difficult to be evaluated due to the complex biological and statistical interrelationships among different risk factors. However, the management of associated risk factors may favourably influence the risk profile in each patient suffering from PAD.


[Clinical significance of serum CA130 level in women with spontaneous abortion, hydatidiform mole and early normal and tubal pregnancy]. We examined the serum CA130 level in women in early pregnancy with normal (n = 13), and abnormal (n = 28) course. The abnormal course consisted of intrauterine spontaneous abortion (n = 10), hydatidiform mole (n = 3) and tubal pregnancy (n = 15). The serum CA130 level was higher than for normal nonpregnant women (35u/ml) in 69% (9/13) of women with normal pregnancy, 90% (9/10) of those with intrauterine spontaneous abortion and 100% (3/3) of those with hydatidiform mole; the mean value and standard deviation for these three groups were 131 +/- 150u/ml (n = 13), 197 +/- 253u/ml (n = 10), and 47 +/- 15u/ml (n = 3), respectively. In contrast, the serum CA130 level of the 15 women with tubal pregnancy was 28 +/- 21u/ml. Among these patients, all of the 13 women without genital bleeding had a CA130 level within the normal range for nonpregnant women (mean +/- SD; 20 +/- 6u/ml). Since CA130 is abundant in the decidual tissue but is scant in tubal tissue, a high CA130 level in maternal sera during early pregnancy may indicate the presence of the destruction of decidual tissue, while a low or normal CA130 level throughout early pregnancy is regarded as characteristic of tubal pregnancy.


[Screening for gestational diabetes mellitus based on blood glucose 2hrs value after a 75g oral glucose load]. We examined the relationship between the time of blood collection and the blood glucose value by analyzing blood glucose values around one hour and two hours after a 75g oral glucose load in 50 women who were at 24 to 28 weeks of gestation. We performed a 75g, 2hrs screening test at 16 to 33 weeks of gestation on all 2,570 pregnant women who visited our out patient clinic between May, 1986 and August, 1991. 1. During the 50-70 minutes after glucose loading, the blood glucose level changed 15-20mg/dl every 10 minutes. During the 110-130 minutes after glucose loading, the change was smaller (10-15mg/dl). These results suggest that the blood glucose level is more stable at 120 minutes than at 60 minutes. 2. When the cut-off point was set at 130mg/dl, the screening test for GDM with a 75g, 2hrs value had a sensitivity of 100%, and a specificity of 85.5% and the incidence of GDM was 3.1%. A 75g, 2hrs screening test therefore seems to be as useful in detecting GDM as a 50g, 1-hour test and to be more suited to crowded out patient clinics because its ability to detect GDM is not seriously affected by small variations in the blood sampling time.


Venous thromboangiitis associated with regional enteritis. Thromboembolic complications occurring in patients with Crohn's disease are increasingly reported and are associated with a high mortality. The mechanism by which the thrombogenic process occurs is unclear. As recent findings suggest that Crohn's disease is a chronic vasculitis with multifocal gastrointestinal infarctions secondary to an imbalance of the cellular hemostatic pathway, extradigestive thrombotic complications might be mediated by the same vascular immune reaction. We report an unusual case of recurrent venous thrombosis associated with regional enteritis, the main point of interest being two-fold: first, this case provides a morphological evidence for an angiitic process as the cause of extradigestive thrombosis; second, the gastrointestinal disease remained subclinical for more than four years although the patient developed major venous thrombotic complications.


Peripheral vascular disease in newly diagnosed non-insulin-dependent diabetic. The aim of this study was to assess the prevalence of peripheral vascular disease (PVD) in newly diagnosed diabetic patients and the possible relationship to various risk factors. One hundred and twenty non-insulin-dependent diabetics (NIDDs) aged 50-70 years and 93 non-diabetic subjects, matched for age and sex, were studied using Doppler ultrasound. None had a history of alcoholic abuse, while 12 diabetic and 8 non-diabetic subjects were smokers. There were 6 male subjects with PVD (5 NIDDs, 1 control subject) and 2 female diabetic subjects with PVD (p: No SD). In group of male diabetics with PVD, HDL-C levels were found to be lower and triglyceride levels higher, than in those without diabetes, but the difference was not significant. Hypertension, body mass index and smoking were not associated with the presence of PVD in either female or male diabetic subjects. It is concluded that, although PVD tended to be more common in men with newly diagnosed diabetes, the overall findings support the view that macrovascular disease is related to duration of diabetes.


[Influences of magnesium sulfate on maternal calcium metabolism in preterm labor]. The effects of intravenous magnesium sulfate tocolysis on calcium metabolism were studied in 10 patients with preterm labor. A loading dose of magnesium sulfate (4g) was administered intravenously maintenance intravenous infusion of magnesium sulfate (1g per hour). All patients simultaneously received 50 micrograms ritodrin per minutes by intravenous infusion. Serum magnesium increased from 1.91 +/- 0.06mg/dl to 4.6 +/- 0.71mg/dl at 30 minutes (p < 0.01) and it remained relatively high. The fall in serum calcium corrected by serum total protein was most rapid during the first 30 minutes, from 9.04 +/- 0.47mg/dl to 8.3 +/- 0.27mg/dl (p < 0.01). Urinary excretion of magnesium, represented as the calcium/creatinine ratio, rose markedly from 0.05 +/- 0.01 to 3.18 +/- 0.8 at an hour (p < 0.01) and thereafter remained higher than the baseline level. Changes in urinary excretion of calcium paralleled those of urinary evcretion of magnesium. Serum parathyroid hormone rose from 118 +/- 42.2pg/ml to 294 +/- 121pg/ml at 6 hours (p < 0.05). Serum 1 alpha,25-(OH)2D3-rose from 89.3 +/- 44.2pg/ml to 126 +/- 38.7pg/ml (p < 0.05). Serum calcitonin showed no significant change. These findings indicate that correction of hypocalcemia mainly depends on secretion of parathyroid hormone in the early stage, and thereafter depends on the cooperative action of parathyroid hormone and 1 alpha,25-(OH)2D3.


[Etiological considerations of the loose shoulder from a biochemical point of view--biochemical studies on collagen from deltoid and pectoral muscles and skin]. In order to elucidate the etiology of the loose shoulder, biochemical characteristics of deltoid and major pectoral muscles and skin collagen from patients with loose shoulder have been investigated. Amounts of soluble collagen, reducible cross-links of collagen and their precursors in both muscles and skin from the loose shoulder were found to be higher than those in the control. These data suggest that soft tissues, especially deltoid muscles, of patients with loose shoulder contain relatively immature collagen fibers compared with normal shoulders. In re-operation cases of the loose shoulder, the collagen fibers in skin were more immature, which may reflect the clinical features of the disease.


[An experimental study on tissue induction in anterior cruciate ligament reconstruction with the scaffold-type polyester artificial ligament (Leeds-Keio)]. The effect of experimental autogenous tissue implantation (infrapatellar fat pad, fascia lata) together with scaffold type artificial ligament (Leeds-Keio) in ACL reconstruction was studied in 56 mongrel adult dogs. Specimens were examined macroscopically, histologically and by means of microangiography. The results were compared with those in which the artificial ligament was used alone. When infrapatellar fat pad was transferred on artificial ligament, tissue induction was observed 2 weeks after reconstruction. When ACL was reconstructed using a combination of artificial ligament and a small piece of fascia lata, maturated dense collagen fibers were observed 24 weeks after reconstruction. These results indicated that quicker tissue induction could be obtained in ACL reconstruction using an artificial ligament together with a infrapatellar fat pad, and that collagen fiber maturation was accelerated when ACL was reconstructed using an artificial ligament together with a small piece of fascia lata.


[Experimental study on the stability of cementless porous coated endoprosthesis]. This study investigated cementless fixation of titanium porous coated endoprosthesis using canine weight-bearing model. Though immature and thin, ingrown trabeculae reached the deepest region of porous coating four weeks after implantation, and gradually matured and thickened thereafter. Pull-out tests of the porous stem at eight weeks showed excellent fixation with average ultimate pull-out force of 142 kgf and average ultimate shear strength of 2.6 GPa. Bone ingrowth rate gradually increased at the proximal and distal region of the stem and, conversely, it gradually decreased at the middle level, suggesting the dependence of bone ingrowth upon the distribution of the amount of stress transferred from implant to the femur. Severe bone remodeling was found on some specimens in which stress transfer was not physiological. Porous coated femoral stems achieve excellent fixation by bone ingrowth, but require physiological stress transfer in order to avoid significant bone remodeling after implantation.


[A histo-morphometric study on peripheral nerve allografts in rats with cyclosporin A]. This investigation evaluates regeneration across peripheral nerve allografts in minor mismatch rats immunosuppressed with Cyclosporin A (CSA). Lewis(RT1(1)) rats were recipients of 20 mm sciatic nerve grafts from allogenic Fischer (RT1(1)) donors. The recipients were randomly allocated to CSA immunosuppressed or untreated groups. CSA was administered at a daily dose of 5 mg/kg by subcutaneous injection for 8 weeks. Two animals from each group were sacrificed at 2, 4, 6, 8 and 12 weeks after the operation, and bilateral sciatic nerves were resected. Until the 8th week, the CSA-treated group showed good vascularization and minimal scar formation. The regeneration was faster and better in the CSA-treated group than in the untreated group. At the 12th week, however, the CSA-treated group showed scarring in the grafted nerves and Wallerian degeneration in the distal nerves, whereas the untreated group showed increased vascularization and myelinated fibers. The results have demonstrated that CSA greatly facilitates the regeneration process across the nerve allograft whereas the discontinuation of CSA leads to reduction of the regenerated nerve fibers. These findings would indicate that the use of CSA is imperatively needed even in minor mismatch cases.


Effects of catechol compound administration on nerve growth factor synthesis in the peripheral nervous system. The effects of the intraperitoneal administration of catechol compounds on nerve growth factor (NGF) synthesis in the peripheral nervous system were examined in Wistar male rats. Five injections of 4-methylcatechol (4-MC) during a three-day period caused a threefold to fourfold increase in the NGF content of organs with sympathetic innervation (heart, submandibular gland) and the sciatic nerve. Next, we investigated time-dependent changes in the NGF content after a single injection of 2 micrograms of 4-MC. A transient increase in the NGF content was detected in the heart and submandibular gland at 16 hours after administration. In the sciatic nerve, a transient increase in the NGF content was noted at 20 hours in the nerve segments on the distal side, and at 24 hours in the segments on the proximal side. In the superior cervical ganglion and dorsal root ganglion, i.e., the locations of the sympathetic and sensory nerve cell bodies, an increase in the NGF content was detected between 32 and 40 hours. Therefore, catechol compounds stimulate NGF synthesis in the peripheral nervous system, and NGF induced by 4-MC is transported retrogradely in the axon to the soma in a physiological manner. Furthermore, in the dorsal root ganglion and superior cervical ganglion, the use of substance P and tyrosine hydroxylase activity as biochemical markers of sensory and sympathetic neurons has demonstrated the biological activity of newly-synthesized NGF induced by catechol compounds.


[An experimental study on direct nerve implantation into denervated muscle--the functional recovery and the site of implantation]. The functional recovery of the newly formed endplates in the muscle reinnervated by direct neurotization was studied grossly, electrophysiologically and histologically in the anterior tibialis muscle of rats. The proximal stump of the tibial nerve was severed at the ankle level and was embedded at the level of distal one fifth of the muscle where no endplates were detected just after denervation of the muscle. Histologically, the accumulations of acetylcholinesterase activity were detected 4 weeks after neurotization. Electrophysiological study using a multi-channel electrode revealed the two-directional propagation of action potentials 8 weeks after neurotization, and the propagation started from no other sites than the nerve-implantation. The muscle tension revealed 42% of the contralateral muscle 52 weeks after neurotization. These results concluded that the function of the newly formed endplates spread throughout the muscle and it was lasting.


[Experimental study on the spinal lesions in hyperostotic mouse (twy/twy): special reference to the pathogenesis of the ossification of the spinal ligaments and to the action of ethane-1-hydroxy-1, 1-diphosphonate (EHDP)]. The hyperostotic changes in the spine of the twy mouse and the action of ethane-1-hydroxy-1, 1-diphosphonate (EHDP) on those hyperostotic changes were studied histomorphometrically. The periosteal bone formation and the endosteal bone resorption in the anterior cortex of the vertebral bodies of the twy mouse continued even after maturation, resulting in the anterior cortex shifting to the anterior direction. In parallel with these changes, calcification in the enthesis (to which spinal ligaments and the outer layer of disc fibrosis are attached) continued, ending in ankylosis of the whole spine. It was similar to a membranous ossification. Moreover calcification in these regions was most sensitively inhibited by EHDP and reappeared as soon as the discontinuation of EHDP. These findings showed that the pathogenesis of the twy mouse was the hyperostosis which continued even after maturation in the genetical region of a membranous ossification, especially in the enthesis. Further they appeared to be similar to the initial changes in ankylosing spinal hyperostosis or the ossification of the spinal ligaments. In addition, our results provided evidence for the usefulness of EHDP in the twy mouse.


[Radiological changes in the femoral side after cementless total hip arthroplasty]. We investigated radiological and functional changes of the femurs with cementless total hip arthroplasty. Our particular attention was paid to the relationship between the thigh pain and the rate, period of its incidence and the progression of radiological changes. The cases studied consisted of 57 female patients with severe coxarthrosis who were implanted with a SUS316-L macroanchoring type of THP (JIAT). Clinical evaluations, radiographic findings in the femoral side, and osteometric observations were carried out periodically in the period before surgery until five years after surgery. The JOA criterion at the time of the final examination was average 91 points. However, we observed a high degree of bone change in the femur on the radiogram. Of 63 hips, there was 71% cortical bone atrophy, a 43% reduction of cortex thickness, 25% cortical bone hypertrophy, 94% endosteal bone bridging in the stem tip, a 64% linear sclerosis of the neighboring stem, and a 37% enlargement of the medullary cavity. Bone atrophy was manifested soon after surgery, and there was 34% recovery. Further, there was no connection between thigh pain and bone changes. We, therefore, concluded that such bone changes as atrophy would not immediately bring about adverse consequences.


Tissue reactions at the implant-bone interface of the cementless total knee prostheses with sinking (Okayama University Mark-II type). Periprosthetic tissues of 14 revised knees of 11 patients, who had undergone Mark-II type cementless total knee prosthesis, were examined to find the relationship between tissue and sinking. Nine patients had rheumatoid arthritis and two had osteoarthritis. Two types of tissue reactions were observed at the implant-bone interface: 1) granulation-dominant tissue, composed of giant cells and macrophages phagocytizing high density polyethylene flakes and dense granules of metal, on the femoral side in six cases, and tibial side in four, 2) fibrosis-dominant tissue on the tibial side in seven, and femoral side in four. On each type, three zones were observed. In the deep zone adjacent to the bone, hyaline cartilage-like tissue was located on the tibial side in four cases, and femoral side in one. An immunohistochemical stain with S-100 proteins and transmission electron microscopy revealed chondrocyte-like features. Chondrogenesis was more frequently observed on the tibial side.


An evaluation of the discrepancy definition of dyslexia. The purpose of this study was to determine whether children with dyslexia, that is, children whose reading levels were significantly lower than would be predicted by their IQ scores, constituted a distinctive group when compared with poor readers, that is, children whose reading scores were consistent with their IQ scores. The performance of children with dyslexia, poor readers, and normally achieving readers was compared on a variety of reading, spelling, phonological processing, language, and memory tasks. Although the children with dyslexia had significantly higher IQ scores than the poor readers, these two groups did not differ in their performance on reading, spelling, phonological processing, or most of the language and memory tasks. In all cases, the performance of both reading disabled groups was significantly below that of nondisabled readers. The findings were similar whether absolute difference or regression scores were used. Reading disabled children, whether or not their reading is significantly below the level predicted by their IQ scores, experience significant problems in phonological processing, short-term and working memory, and syntactic awareness. On the basis of these data, there does not seem to be a need to differentiate between individuals with dyslexia and poor readers. Both of these groups are reading disabled and have deficits in phonological processing, verbal memory, and syntactic awareness.


Academic achievement following childhood brain disease: implications for the concept of learning disabilities. The present study examined three hypotheses regarding the consequences of early brain damage for academic achievement: First, early brain insults will have a negative impact on achievement, even in children with normal intelligence. Second, underachievement in these children will be at least partially independent of IQ (i.e., not fully accounted for by a lowering of IQ within the average range). Third, normally intelligent children with histories of brain insult will also manifest selective cognitive dysfunctions. To test these hypotheses, we compared two groups of children who had recovered from Haemophilus influenzae type b meningitis. The "complicated" group consisted of children who, as a consequence of having sustained neurologic complications during their illness, were more likely to have had brain insults. Children in the "uncomplicated" group did not have complications with their illness and were regarded as having escaped significant central nervous system (CNS) pathology. Only children with normal hearing and a prorated Full Scale IQ of at least 80 (WISC-R) were considered. Group differences on the Wide Range Achievement Test-Revised were consistent with the first two hypotheses. Although the two groups had similar Verbal IQs, the complicated group also had a lower mean Performance IQ and performed less well on perceptually demanding neuropsychological tasks. Findings suggest that learning disabilities may have selective, brain-related cognitive antecedents; but they challenge the practice of using IQ criteria for clinical diagnosis.


Discrepancy compared to low achievement definitions of reading disability: results from the Connecticut Longitudinal Study. We used data derived from a survey sample, the Connecticut Longitudinal Study (CLS), to compare two commonly employed definitions of reading disability: a discrepancy-based model (D) and a low reading achievement model (L). We identified children satisfying each definition in second grade and compared the groups retrospectively in kindergarten and prospectively in fifth grade using parent-based, teacher-based, and child-based measures. Our findings suggest more similarities than differences between the reading disabled groups. The most salient differences were those related to ability and seem inherent in the definitions of the groups: Children identified as D have significantly higher verbal, performance, and full scale IQ scores than those identified as L. These findings suggest that both groups of children with reading disability, that is, those defined by either D or L, should be considered eligible for special education services.


Minor neurological dysfunction is more closely related to learning difficulties than to behavioral problems. In a group of 570 nine-year-old children (315 boys and 255 girls) without an overtly handicapping neurological condition, relationships were studied between the presence of minor neurological dysfunction (MND) on the one hand and cognitive and behavioral problems on the other hand. The aim of the study was to investigate whether MND was more closely related to learning problems than to behavioral difficulties. The group was a subpopulation of the birth cohort of the Groningen Perinatal Project in the Netherlands, in which perinatal developmental relationships are studied. The age-specific and standardized neurological examination technique of Touwen (1979) was used, resulting in a description of the neurological condition in terms of the number of deviant clusters. The absence of deviant clusters indicated a normal neurological condition (n = 418); children with one or two deviant clusters were classified as MND-1 (n = 86) and those with more than two deviant clusters as MND-2 (n = 66). The presence and severity of MND was significantly related to poor performance on standardized reading, spelling, and arithmetic tests. MND was also related to ratings of distractible behavior on parent and teacher questionnaires, but to a lesser extent than the cognitive problems. Ratings of so-called "troublesome" and "timid" behavior were not related to MND. Our conclusion is that learning problems are more closely related to MND than are behavioral difficulties. This has implications for prevention and intervention: In the former the focus should be more on biological hazards, in the latter on environment and rearing attitudes.


Comparison of visual-spatial performance strategy training in children with Turner syndrome and learning disabilities. This study examined the effects of a verbal mediation strategy on three groups of subjects who had visual-spatial deficits. Thirteen females with Turner syndrome, 13 females with nonverbal learning disabilities, and 14 males with nonverbal learning disabilities, who ranged in age from 7 to 14 years, were taught via a cognitive behavioral modification approach to verbally mediate a spatial matching task. Pretest and posttest performance differences on parallel forms of a visual-spatial orientation task were examined. All three groups showed significant improvement in visual-spatial task performance after the training. There were no significant differences in the degree of improvement among the three groups. The results suggest that children with Turner syndrome may benefit from problem-solving strategy training in a manner similar to children with nonverbal learning disabilities.


Undergraduate admission policies, practices, and procedures for applicants with learning disabilities. This study examined (a) the academic and nonacademic criteria used by admission personnel to determine the eligibility of undergraduate applicants with learning disabilities, (b) agreement of criteria used by institutions of varying competitiveness, and (c) the frequency with which admission personnel conduct validity studies on these criteria. A nonrandom sample consisting of 66 state universities and colleges in the Northeast was surveyed. The results suggest that the academic and nonacademic criteria employed are similar to those employed for applicants without learning disabilities, the criteria used by different types of institutions differ significantly, and admission personnel do not conduct validity studies. Further clarification and validation of admission criteria appears warranted.


Similarities in the social competencies of learning disabled and low achieving elementary school children. This study sought to examine distinctions in social competencies between children with learning disabilities (LD) and other children who also experience academic difficulties. Eighty-five children with LD (54 male, 31 female) in Grades 3 through 6 from a large urban school district were compared to a group of low achieving (LA) peers matched on achievement as well as sex, race, and grade. The samples were 42% black, 39% Hispanic, and 19% Anglo. Both groups completed two self-concept questionnaires, a loneliness scale, and a measure of their social relationships outside of school. In addition, their classmates completed a peer rating scale and their teachers completed two ratings of the child's social skills. The results indicated that children with LD and LA children were comparable on most measures, although children with LD reported themselves as being less lonely than LA children. In addition, regular-class children rated children with LD as more likable than LA children. The results highlight similarities in the social competencies of children with LD and LA children and suggest that special education classes may offer some social advantages to children with mild handicaps.


Students with learning disabilities in the university environment: a study of faculty and student perceptions. This study examines faculty and student perceptions regarding university students with learning disabilities, sensitivity to such students' special needs, accommodations, and the perceived impact of a learning disability. Results reveal a general sensitivity to the special needs of students with learning disabilities; however, group differences suggest several areas warranting further attention.


Identifying alterable patterns in employment success for highly successful adults with learning disabilities. Adults with learning disabilities were studied to ascertain the patterns of successful functioning that promoted high levels of vocational success. This area of research has been neglected in the developing research base on adults with learning disabilities. In this study of 46 highly successful and 25 moderately successful adults with learning disabilities using ethnographic interviews, it was found that the overriding theme was control and that control was sought through the pursuit of two sets of themes--internal decisions and external manifestations. These themes transcended the entire sample, and the clear difference between the groups was the degree of attainment on the various elements the themes comprised. These elements and themes are discussed and a model of successful vocational functioning is developed and explained.


The neurobiology and neuropsychology of adult learning disorders. The area of adult learning disabilities is reviewed from a neurobiological and neuropsychological perspective. The seminal contributions of Galaburda's neuroanatomic studies, Duffy's neurophysiologic findings, and recent magnetic resonance imaging studies have illustrated various possible pathophysiologic bases in dyslexia and are reviewed along with other neurobehavioral disorders that may coexist with learning disorders. Neuropsychological assessment methods in evaluating learning disabilities are reviewed and test results in a case of traumatically induced dyslexia are compared to those obtained in three adults with residual developmental dyslexia. The problems associated with treatment of adult learning disorders are discussed in the context of brain plasticity and recovery of function from the perspective of a brain-injury model. Lastly, the basis common to neurologic factors subserving learning disability and emotional dysfunction is explored.


Adults with learning disabilities: current and future research priorities. This article presents a review of current research, or what is currently known about adults with learning disabilities. The review is organized under different settings, including the community, postsecondary, and employment environments, and is followed by a review of longitudinal studies. Based on the review of the current status of the literature, research priorities for the 1990s are presented, including (a) identifying and conducting research with specific individuals or samples within the adult (LD) population; (b) identifying successful intervention techniques within the cognitive, social, personal, and vocational domains; (c) conducting research in a variety of environments, including community, employment, and postsecondary settings and across critical periods of an individual's lifespan (such as movement from postsecondary environments into employment and from employment to retirement); and (d) examining alternative methodologies, such as single-subject, case study, group research, and so forth, in future investigations. The priorities are followed by a description and comparison of various research methodologies.


The relationship of phonological awareness, rapid naming, and verbal memory to severe reading and spelling disability. The present study examined the relationship of phonological awareness, naming speed, and verbal memory to the scores obtained from five tests assessing word attack, word identification, reading comprehension, and spelling skills in 54 children with severe reading disabilities (48 boys and 6 girls; M age = 9 years, 7 months). Multiple regression analyses indicated that the best predictor of achievement across the five academic tests was the Verbal Comprehension factor from the Wechsler Intelligence Scale for Children-Revised. Age, socioeconomic status (SES), and externalizing behavior problems were also significant predictors of achievement, depending on the academic measure. After controlling for age, SES, behavior problems, and intelligence, the phonological awareness task added significantly to the prediction of word attack, spelling, and reading comprehension scores; rapid letter naming added significantly to the prediction of word identification and prose passage speed and accuracy scores; and a word-list memory task added significantly to the prediction of word recognition scores. These results suggest that several independent processes interact to determine the extent and severity of reading problems.


The perspectives of mothers whose children are in special day classes for learning disabilities. This article presents information about the attitudes and beliefs of mothers concerning their children in learning disabilities classes. Four mothers were interviewed over a 1-year period. Their comments about services provided by educational and mental health professionals indicated high levels of frustration and dissatisfaction. Parental expectations and perspectives differed from those of educators. Participants and educators were found to view learning disabilities differently.


Information flow in the hospital: a comparative study of the Hungarian and the Dutch situation using a two axes model of hospital information flow. This study compares the organization and structure of information flow in a Dutch and in a Hungarian hospital. The study was carried out as a field orientation part of a Health Care Management project of the Hogeschool van Amsterdam. The host of the field orientation was the BAZIS Foundation, the Central Development and Support Group Hospital Information System, Leiden. The visited hospitals were equipped with the BAZIS Hospital Information System. The method of study consisted of series of formalized interviews with all-level actors of a hospital; the interpretation of data was enhanced by a two axes (patient and management) model of information flow defined by the authors. In summary, Dutch hospitals show a more elaborate information flow system, with more information flow channels sideways among equal levels, less bureaucracy in organization of information flow, and significantly more benefits of automation, compared to the Hungarian situation.


Standardized nursing language for healthcare information systems. Since a substantial component of health care delivery is reflected in nursing's work, it is imperative that nursing expedites implementation of a standardized language that reflects nursing's work and ultimately allows outcome evaluation. This paper will summarize the state of development and related issues of standardized language in nursing, including: Nursing Minimum Data Set, Taxonomies of Nursing Diagnoses, Nursing Interventions, Outcomes, and the Nursing Management Minimum Data Set. The Nursing Minimum Data Set, including nursing care, patient or client demographic, and service elements, reflects a standardized collection of essential nursing data used by multiple data users in the health care delivery system across all types of settings. The nursing care elements include nursing diagnosis, nursing intervention, nursing outcome, and intensity of nursing care. Currently, more than 100 nursing diagnoses have been accepted for clinical testing by the North American Nursing Diagnosis Association (NANDA) and have been incorporated into a taxonomy of nursing diagnoses that reflects patient responses to actual or potential health problems that nursing can address. A current formulation of a taxonomy of nursing interventions for the treatment of the nursing diagnoses yielded 336 nursing intervention labels organized at three or four levels of abstraction. Concomitant with these endeavors is the necessity for identifying outcomes associated with each diagnosis and its treatment. Concepts and a classification for indicators of these outcomes are being reviewed. Last, to address the contextual covariates of patient outcomes, a collection of core variables needed by nurse managers to make management decisions and compare nursing effectiveness across institutions and geographic regions is under development. In summary, standardized measures to determine cost effective, high quality, appropriate outcomes of nursing care delivered across settings and sites are being developed.


A visual field quantification system for the Goldmann Perimeter. We have developed a visual field quantification system that can accurately quantify the measurement results by the Goldmann Perimeter (GP). This system calculates the plural indexes introduced in published papers. In recent years, several isopter quantification methods have been proposed. In these methods, the isopters are digitized and the data are input into a computer, after which a computer program evaluates the changes in the isopters quantitatively. However, each program is only able to evaluate private indexes. We have developed a system that quantifies data using multiple methods. This system used five methods that have been introduced in published papers. With this system, a physician can analyze GP data with multiple methods and can diagnose diseases accurately. We have already input about 2500 GP recording papers into the computer by this system. We then calculated the plural indexes, and visualized the temporal changes in the perimetry.


The effect of a multiple literature database search--a numerical evaluation in the domain of Japanese life science. In literature database searching, we show that it is necessary to use plural databases for a more improved search. We also compare the results of a single database search with that of multiple database search in the domain of Japanese life sciences. We searched the MEDLINE and EMBASE using the same search terms. There were some differences in the results, owing to differences in the journals and recording methods. We herein show some of the differences in the journals contained in both databases. Furthermore, we show the differences in the number of papers derived from the same journal. Next, as an example of a practical search, we selected some universities in Japan, searched both databases regarding papers published from these universities and then merged the results by hand. According to our results, only 63% of all papers were common to both databases.


The prevalence and risk factors of hypertension in population aged 30-65 years in rural area, Amphoe Phon, Khon Kaen. The prevalence of hypertension in a population aged 30-65 years in a rural area in Amphoe Phon, Khon Kaen was 18 per thousand (22 per thousand in males and 16 per thousand in females). The prevalence of isolated systolic hypertension was 5 per thousand (10 per thousand in males and 2 per thousand in females). The risk factors of blood pressure elevation were age, body mass index, level of development of village and non farmer occupation.


Liver injuries: a five-year study at Surat-thani Hospital. Most liver injuries can usually be handled by simple technique. In this study, 70 per cent of the cases needed only simple suture. There were 10 cases (16.67%) in which no surgical procedures were required at all, because the bleeding had stopped prior to surgery. Complications and mortality varied according to the type of liver injury, the involved organ injury and the most important factor, shock prior to and during surgery. This study reports not only the type of liver injury, causes, treatment, complications and mortality rate but it also compiles the surgical techniques used in managing liver injury according to severity.


Anterior lumbar discectomy: a study of iliac and fibular bone graft. The author studied the iliac and fibular autogenous bone grafts in anterior lumbar discectomy for herniated disc of 29 patients. They were examined 2 1/2 to 8 years postoperatively. Iliac numbness was found in 4 of 15 patients (26.7%) and fibular pain was found in one of 14 patients (7.1%). For functional result, iliac and fibular graft were found excellent in 86.6 and 92.9 per cent respectively. There was no statistically significant difference in proportion of patients experiencing discomfort at donor site or in the functional result between iliac and fibular grafts.


A comparative study of isobaric and hyperbaric solution of bupivacaine for spinal anaesthesia in caesarean section. Though hyperbaric solution of bupivacaine following intrathecal injection had satisfactory spread of analgesia, it regressed rapidly with more side effects. Isobaric bupivacaine seemed to provide a slow regression of analgesia with fewer undesirable effects except for its inadequate spread of analgesia. As a result, if the dosages as well as the time of administration of isobaric solution are well adjusted, we believe that it is safe and reliable with an excellent level of analgesia for caesarean section.


In vitro antimicrobial activity of cefodizime, a third generation cephalosporin. Cefodizime is one of the new broad-spectrum cephalosporins. It is an aminothiazolyl iminomethoxy cephalosporin which is metabolically stable and has a prolonged serum half life. Cefodizime was primarily active against gram-negative bacilli and at the concentration of 0.5 mg/L, it inhibited 90 per cent of Enterobacteriaceae. P. mirabilis was the most susceptible species tested (MIC90 of 0.02 mg/L). E.coli, K.pneumoniae, Salmonella spp., Shigella spp. and M. morganii were also very sensitive to cefodizime, with the MIC90 of 0.25-0.5 mg/L. Cefodizime, however, was not active against most gram-negative bacilli possessing Type I beta-lactamases of Richmond and Sykes, namely, Enterobacter spp., P. aeruginosa and A. anitratus (MIC90 of > 128 mg/L). Among gram-positive bacteria, only S.pyogenes was highly susceptible (MIC90 of 0.05 mg/L), while S. aureus (methicillin-sensitive) was moderately susceptible and Enterococcus spp. was resistant. Cefodizime appeared to be bactericidal and was not affected by serum. High inoculum (10(7) cfu/ml) of K.pneumoniae and Enterobacter spp. resulted in increase of the MIC of cefodizime. This study shows that local bacterial isolates in a university hospital in Bangkok, Thailand were not different in susceptibility pattern from those reported in developed countries. The in vitro activity of cefodizime as a third generation cephalosporin, with its good pharmacokinetic property, and the property of the agent as a biological response modifier, should prove that this is a promising new agent in treating serious infections especially in immunosuppressed hosts.


HIV infection in male patients attending a sexually transmitted disease clinic. Three hundred and fifty-two heterosexual males, attending a sexually transmitted disease clinic at Siriraj Hospital from December 1989 to February 1991 were studied for the prevalence of HIV infection. Of these, 334 men reported prostitutes as the main source of their sexually transmitted disease. No one had received blood transfusion in the last 5 years, and there was no history of intravenous drug use, homosexuality or bisexuality. HIV antibody was found in the sera of 24 men (6.8%). HIV seropositivity was associated with serologic makers of syphilis (P < 0.05) but was not associated with present genital ulcers on physical examination or other STDs. These data indicate the high rate of female prostitutes to male transmission of HIV infection in the presence of sexually transmitted disease and confirms the relationship between syphilis and HIV infection. HIV/AIDS educational programmes and campaigns to promote condom use among prostitutes and clients are an urgent need in Thailand.


Primary lymphoma of the esophagus: a case report. A case of primary non-Hodgkin's lymphoma of the esophagus was reported. The macroscopic manifestation from esophagography, computed tomography and esophagoscopy were demonstrated. Pathological findings were demonstrated. The tumour responded well to combined therapy of surgery, radiotherapy and chemotherapy.


Acute cyanide poisoning: a case report with toxicokinetic study. Cyanide poisoning is a life threatening condition. But specific antidotes exist and can be easily prepared from available substances in hospital. Administration of antidotes will produce methemoglobin, which itself causes hypoxia. Nitrite induced methemoglobin can be extremely dangerous and even lethal. Before administering the antidotes, the diagnosis should be confirmed. Nitrite should not be given if the poisoning is mild or diagnosis is uncertain, to avoid excessive methemoglobin, dosage of sodium nitrite must be adjusted according to hemoglobin level (Table 1). Usage of sodium nitrite and sodium thiosulfate in the recommended doses are safe and effective for cyanide poisoning.


Granulomatous mastitis: case report and review of literature. A 38-year-old woman presented with a 5-month history of a right breast mass after delivery of her second child. The lesion simulated the picture of a carcinoma on clinical grounds. Fine-needle aspiration also reported an erroneous diagnosis of malignancy. At surgery, a discrete granuloma with microabscesses was seen without any causative organisms being identified. Open biopsy or Tru-Cut needle biopsy is recommended to prevent such an error which may result in unnecessary radical surgery.


Activation of the classical complement pathway by mannose-binding protein in association with a novel C1s-like serine protease. Serum mannose-binding protein (MBP) is a C-type lectin that binds to terminal mannose and N-acetylglucosamine moieties present on surfaces of certain pathogens and activates the classical complement pathway. In the present study, we describe the mechanism underlying the activation triggered by MBP. The human serum MBP fraction was obtained by sequential affinity chromatography on mannan-Sepharose, anti-IgM-Sepharose and anti-MBP-Sepharose in the presence of calcium ions. This fraction contained a C1s-like serine protease as assessed by C4 consumption. The C1s-like serine protease, designated MBP-associated serine protease (MASP), was separated from MBP by rechromatography on anti-MBP-Sepharose in the presence of ethylenediaminetetraacetic acid. MASP exhibited both C4- and C2-consuming activities. The molecular mass of MASP was estimated to be 83 kD with two polypeptides of heavy (66 kD) and light (L) (31 kD) chains linked by disulfide bonds. The serine residue responsible for protease activity is located on the L chain. Reconstitution experiments using MASP and MBP revealed that combination of the two components restores C4- and C2-activating capacity on mannan. Based on analyses of molecular size, antigenicity, and 11 NH2-terminal amino acid sequences of the L chain, we conclude that MASP is a novel protein different from C1r or C1s. Our findings are not in accord with a proposed mechanism by which MBP utilizes the C1r2-C1s2 complex to initiate the classical complement pathway.


Human lymphokine-activated killer cells are cytotoxic against cells infected with Toxoplasma gondii. Experiments were conducted to determine whether human lymphokine-activated killer (LAK) cells are cytotoxic against cells infected with Toxoplasma gondii. Nylon wool nonadherent (NWNA) peripheral blood lymphocytes, as well as purified natural killer cell (NK) (CD3- CD16+ CD56+) and T (CD3+ CD16- CD56-) cells obtained from five healthy T. gondii seronegative volunteers exhibited minimal cytotoxic activity against T. gondii-infected cells. When standard LAK (S-LAK) cell preparations were induced by incubation of NWNA cells with recombinant interleukin 2, induction of remarkable cytotoxic activity against T. gondii-infected cells. When standard in LAK cell preparations from each of the volunteers. The phenotype of the LAK precursor and effector cells varied depending on the target cell used. Whereas the precursor and the effector cells of most of the LAK activity against K562 and Daudi cells were cells with NK phenotype, when T. gondii-infected cells were used as targets, both cells with NK and T cell phenotypes were precursors and effectors of the lysis. When cytotoxic activity of S-LAK cells was compared with the activity of adherent LAK (A-LAK) cells, A-LAK cells displayed higher cytotoxic activity against T. gondii-infected cells, as well as against K562 and Daudi cells. Cold target inhibition experiments suggested that there is a subset of LAK effector cells capable of lysing both T. gondii-infected cells and Daudi cells, whereas other subsets preferentially or exclusively lyse one of these target cells.


Purification of three cytotoxic lymphocyte granule serine proteases that induce apoptosis through distinct substrate and target cell interactions. We recently reported the purification of a lymphocyte granule protein called "fragmentin," which was identified as a serine protease with the ability to induce oligonucleosomal DNA fragmentation and apoptosis (Shi, L., R. P. Kraut, R. Aebersold, and A. H. Greenberg. 1992. J. Exp. Med. 175:553). We have now purified two additional proteases with fragmentin activity from lymphocyte granules. The three proteases are of two types; one has the unusual ability to cleave a tripeptide thiobenzyl ester substrate after aspartic acid, similar to murine cytotoxic cell protease I/granzyme B, while two are tryptase-like, preferentially hydrolyzing after arginine, and bear some homology to human T cell granule tryptases, granzyme 3, and Hanukah factor/granzyme A. Using tripeptide chloromethyl ketones, the pattern of inhibition of DNA fragmentation corresponded to the inhibition of peptide hydrolysis. The Asp-ase fragmentin was blocked by aspartic acid-containing tripeptide chloromethyl ketones, while the tryptase fragmentins were inhibited by arginine-containing chloromethyl ketones. The two tryptase fragmentins were slow acting and were partly suppressed by blocking proteins synthesis with cycloheximide in the YAC-1 target cell. In contrast, the Asp-ase fragmentin was fast acting and produced DNA damage in the absence of protein synthesis. Using a panel of unrelated target cells of lymphoma, thymoma, and melanoma origin, distinct patterns of sensitivity to the three fragmentins were observed. Thus, these three granule proteases make up a family of fragmentins that activate DNA fragmentation and apoptosis by acting on unique substrates in different target cells.


Comparative clonal analysis of human immunodeficiency virus type 1 (HIV-1)-specific CD4+ and CD8+ cytolytic T lymphocytes isolated from seronegative humans immunized with candidate HIV-1 vaccines. The lysis of infected host cells by virus-specific cytolytic T lymphocytes (CTL) is an important factor in host resistance to viral infection. An optimal vaccine against human immunodeficiency virus type 1 (HIV-1) would elicit virus-specific CTL as well as neutralizing antibodies. The induction by a vaccine of HIV-1-specific CD8+ CTL in humans has not been previously reported. In this study, CTL responses were evaluated in HIV-1-seronegative human volunteers participating in a phase I acquired immune deficiency syndrome (AIDS) vaccine trial involving a novel vaccine regimen. Volunteers received an initial immunization with a live recombinant vaccinia virus vector carrying the HIV-1 env gene and a subsequent boost with purified env protein. An exceptionally strong env-specific CTL response was detected in one of two vaccine recipients, while modest but significant env-specific CTL activity was present in the second vaccinee. Cloning of the responding CTL gave both CD4+ and CD8+ env-specific CTL clones, permitting a detailed comparison of critical functional properties of these two types of CTL. In particular, the potential antiviral effects of these CTL were evaluated in an in vitro system involving HIV-1 infection of cultures of normal autologous CD4+ lymphoblasts. At extremely low effector-to-target ratios, vaccine-induced CD8+ CTL clones lysed productively infected cells present within these cultures. When tested for lytic activity against target cells expressing the HIV-1 env gene, CD8+ CTL were 3-10-fold more active on a per cell basis than CD4+ CTL. However, when tested against autologous CD4+ lymphoblasts acutely infected with HIV-1, CD4+ clones lysed a much higher fraction of the target cell population than did CD8+ CTL. CD4+ CTL were shown to recognize not only the infected cells within these acutely infected cultures but also noninfected CD4+ T cells that had passively taken up gp120 shed from infected cells and/or free virions. These results were confirmed in studies in which CD4+ lymphoblasts were exposed to recombinant gp120 and used as targets for gp120-specific CD4+ and CD8+ CTL clones. gp120-pulsed, noninfected targets were lysed in an antigen-specific fashion by CD4+ but not CD8+ CTL clones. Taken together, these observations demonstrate that in an in vitro HIV-1 infection, sufficient amounts of gp120 antigen are produced and shed by infected cells to enable uptake by cells that are not yet infected, resulting in the lysis of these noninfected cells by gp120-specific, CD4+ CTL.(ABSTRACT TRUNCATED AT 400 WORDS)


Glucocorticoids induce the expression of CD8 alpha chains on concanavalin A-activated rat CD4+ T cells: induction is inhibited by rat recombinant interleukin 4. Rat T lymphocytes, activated in vitro with concanavalin A (Con A), were shown by flow cytofluorographic analysis to contain a population of cells that simultaneously expressed CD4 and the alpha chain of CD8. The inclusion of the glucocorticoid hormone dexamethasone in the culture medium greatly increased both the frequency of these double-positive cells and the level of CD8 alpha chain expression. The level of expression of CD4 was not affected, and the cells that expressed CD8 antigen only also remained unchanged in surface phenotype. Detailed studies demonstrated unequivocally that the CD4+ CD8 alpha + cells were not artifacts produced by the random association of single-positive cells in the flow cytofluorograph, but arose from precursors that were single-positive CD4+ cells before activation. Furthermore, Con A activation of purified CD4+ T cells, in the presence of T cell-depleted accessory cells, showed that CD8+ T cells played no role in the induction process. However, the induction of CD8 alpha chain expression on CD4+ T cells and the enhancement of this expression by dexamethasone were almost completely inhibited by rat recombinant interleukin 4 (IL-4). Detection of mRNA for rat CD8 alpha chain by Northern blot closely paralleled the cell surface expression of CD8 alpha antigen, indicating that dexamethasone and IL-4 had opposing effects on mRNA levels. In contrast, IL-4 and dexamethasone both induced CD8 alpha chain expression on a rat CD4+ T cell clone when this was activated by specific antigen, and, although the effect with IL-4 was relatively weak, it did not antagonize the effect of the glucocorticoid. The possible significance of these results is briefly discussed.


Predominance of fetal type DJH joining in young children with B precursor lymphoblastic leukemia as evidence for an in utero transforming event. The presence of N sequences in the complementarity determining region 3 (CDR3) of the rearranged immunoglobulin H chain is developmentally regulated: N regions are generally present in the DJH joinings of adult B cells but are often absent in fetal B cells. Analysis of the CDR3 in 61 B precursor acute lymphoblastic leukemias indicated that 87.5% of the leukemias obtained from children < or = 3 yr old lacked N regions at the DJH junction. In contrast, in children > 3 yr old, only 11.1% of the leukemias lacked N regions at this junction, a frequency similar to what we have observed in B cells from children and adults. These findings suggest that the majority of leukemias presenting within the first 3 yr of age arise from an in utero transforming event.


A severe combined immunodeficient (SCID) mouse model for infection with Entamoeba histolytica. We used severe combined immunodeficient (SCID) mice to study resistance to invasive infection with Entamoeba histolytica. Seven of seven SCID mice developed liver abscesses when challenged intrahepatically with virulent HM1:IMSS strain E. histolytica trophozoites. Only one of seven similarly challenged immunocompetent congenic C.B-17 mice developed an abscess. Adoptive transfer of polyclonal rabbit anti-E. histolytica antiserum, but not preimmune rabbit serum, completely protected 7 of 12 SCID mice from intrahepatic challenge with ameba. These results demonstrate that lymphocyte-based immunity is important in protection against amebic liver abscess, and that anti-E. histolytica antibody can protect against amebic infection in this system. The SCID mouse may provide a powerful model for studying the components of protective immunity to invasive amebiasis.