Follicular lymphomas of the gastrointestinal tract. Pathologic features in 31 cases and bcl-2 oncogenic protein expression. The gastrointestinal tract is the most common site for extranodal lymphomas, but follicular lymphomas involving the gut are rare. To study their pathologic features and bcl-2 expression, 31 follicular lymphomas of the GI tract were reviewed and unstained paraffin sections from 24 of the cases were immunohistochemically stained using a monoclonal antibody for the peptide product of the proto-oncogene bcl-2. The most common site of lymphoma involvement was the small intestine, especially the terminal ileum. Gastric lymphomas tended to present clinically with symptomatic ulcers and small intestinal lesions presented with obstruction. Five cases involving the terminal ileum or colon had a gross appearance of multitudinous mucosal polyps and were considered to represent examples of "multiple lymphomatous polyposis." Enhanced expression of the bcl-2 oncogenic protein was detectable in lymphoma cells in 75% of cases and at lower levels in normal lymphoid cells in most cases. Small cleaved or mixed cell lymphomas were more likely to show enhanced expression than were large cell cases. Reactive germinal centers showed no bcl-2 staining. It is concluded that follicular GI lymphomas are associated with distinctive pathological features. In their tendency to express bcl-2, these neoplasms resemble their lymph node-based counterparts. Immunohistochemical staining for enhanced bcl-2 expression is of potential diagnostic utility in distinguishing between follicular lymphoma and follicular lymphoid hyperplasia in the gastrointestinal tract. The relevance of the results to lymphoma of mucosa-associated lymphoid tissue (MALT) is discussed.


Phagocytosis and deposition of vascular beta-amyloid in rat brains injected with Alzheimer beta-amyloid. The presence of extracellular deposits of beta-amyloid protein in the brain is a hallmark of Alzheimer's disease (AD). In an effort to determine the effect of amyloid in an animal model, the authors injected amyloid cores isolated from AD brains into the cortex and hippocampus of rats. Lipofuscin, a major contaminant of the plaque core preparation, was injected on the contralateral side and used as a control to induce an analogous phagocytic cell response. Rats were sacrificed 2 days, 7 days, and 1 month after injection and amyloid located by four histochemical techniques. Amyloid and lipofuscin move from the site of injection into otherwise undamaged neuropil, persist for at least 1 month and are both associated with increases in glial fibrillary acidic protein and microglia (OX-42) staining. By 1 week, many of the amyloid cores are ingested by phagocytes. Some of the beta-amyloid-containing phagocytes migrate to the vessels and to the ventricles, and by 1 month, a significant amount of the amyloid is directly associated with the vessels. This suggests that phagocytic cells can internalize exogenous amyloid and attempt to clear it from the central nervous system (CNS). Therefore, the observed distribution of amyloid is not necessarily the initial site of deposition.


Platelet-derived growth factor (BB homodimer), transforming growth factor-beta 1, and basic fibroblast growth factor in dermal wound healing. Neovessel and matrix formation and cessation of repair. Recombinant platelet-derived growth factor (BB homodimer, rPDGF-BB), transforming growth factor beta 1 (rTGF-beta 1), and basic fibroblast growth factor (rbFGF) can accelerate healing of soft tissues. However, little information is available characterizing the components of wound matrix induced by these growth factors and the molecular mechanisms underlying accelerated repair and wound maturation. In this study, the composition, quantity, and rate of extracellular matrix deposition within growth factor-treated lapine ear excisional wounds were analyzed at different stages of healing using specific histochemical and immunohistochemical stains, coupled with image analysis techniques. Single application of optimal concentrations of each growth factor accelerated normal healing by 30% (P less than 0.0003); rPDGF-BB markedly augmented early glycosaminoglycan (GAG) and fibronectin deposition, but induced significantly greater levels of collagen later in the repair process, compared with untreated wounds rTGF-beta 1 treatment led to rapidly enhanced collagen synthesis and maturation, without increased GAG deposition. In contrast, rbFGF treatment induced a predominantly angiogenic response in wounds, with a marked increase in endothelia and neovessels (P less than 0.0001), and increased wound collagenolytic activity (P less than 0.03). rbFGF-treated wounds did not evolve into collagen-containing scars and continued to accumulate only provisional matrix well past wound closure. These results provide new evidence that growth factors influence wound repair via different mechanisms: 1) rPDGF-BB accelerates deposition of provisional wound matrix; 2) rTGF-beta 1 accelerates deposition and maturation of collagen; and 3) rbFGF induces a profound monocellular angiogenic response which may lead to a marked delay in wound maturation, and the possible loss of the normal signal(s) required to stop repair. These results suggest that specific growth factors may selectively regulate components of the repair response by differing mechanisms, offering the potential for targeted therapeutic intervention.


The sequential development of abnormal prion protein accumulation in mice with Creutzfeldt-Jakob disease. The distribution and sequential development of prion protein (PrP) accumulation in the central nervous system (CNS) and non-neuronal organs of mice infected with Creutzfeldt-Jakob disease (CJD) were investigated immunohistochemically using a new pretreatment method that greatly enhanced the immunoreactivity of PrP. Prion protein accumulation in the CNS was first detected at 30 days after inoculation and then developed near the inoculation site or periventricular area, and later spread to the whole cerebrum and then to the pons. Its staining took some characteristic forms. Among non-neuronal organs, PrP accumulated in the follicular dendritic cells (FDCs) in spleen, lymph node, Peyer's patch, and thymus. FDCs staining appeared in spleen, lymph node, and Peyer's patch at 21 or 30 days after inoculation, and in thymus at 90 days. Germinal centers developed in the thymus of some CJD-infected mice. No PrP staining was detected in any examined organs of age-matched control mice.


Mutated c-Ha-ras oncogene alters cytokeratin expression in the human breast epithelial cell line MCF-10A. MCF-10A, a spontaneously immortalized human breast epithelial cell line, has been transformed by transfection with the mutated c-Ha-ras oncogene obtained from T24 carcinoma cells. The pattern of cytokeratin expression was studied in MCF-10A cells in comparison with plasmid transfected or MCF-10Aneo cells, normal ras proto oncogene transfected or MCF-10AneoN cells, and transformed or MCF-10AneoT cells. Cytokeratin expression was studied by western immunoblot of total cellular proteins separated by two-dimensional gel electrophoresis. Blots with cytokeratin specific AE1 and AE3 antibodies showed identical molecular weight species of cytokeratins in MCF-10A, MCF-10Aneo, MCF-10AneoN, and MCF-10AneoT cells; however, in MCF-10AneoT cells, the intensity of immunostaining and the number of immunoreactive phosphorylated polypeptides keratins 7, 8, 15, and 16 was decreased. It was concluded that c-Ha-ras oncogene-induced transformation alters quantitatively the cytokeratin pattern of human breast epithelial cells and that these changes could explain some of the morphologic alterations observed in c-Ha-ras transformed cells.


Cell death in cranial neural crest development. The rhombencephalic neural crest, crucial to the patterning and development of many craniofacial structures, migrates laterally from the dorsal hindbrain, but not as a continuous sheet. We have used a vital dye to demonstrate a discontinuous pattern of cell death in the dorsal midline of the avian rhombencephalon associated with the migration of the neural crest. Whilst cell death commences in the dorsal midline of the presumptive mesencephalon at stage 8, two distinct domains of cell death are apparent in the rhombencephalon by stage 11. The rostral domain lies over primary rhombomere RhA1 and rhombomere rh3, while the caudal domain occurs on the neural midline between the otic vesicles, in the region of rh5. Using a marker for the neural crest, we show that the rostral and caudal domains of cell death correlate with the absence of neural crest migration from rh3 and rh5. Thus segment-specific cell death in the dorsal region of particular rhombomeres may account for their subsequent failure to contribute to the cranial neural crest.


Simian virus 40-induced disease in rhesus monkeys with simian acquired immunodeficiency syndrome. Simian virus 40 (SV40) disease was diagnosed in four rhesus monkeys that died with SIV-induced acquired immunodeficiency syndrome (AIDS). One juvenile monkey seroconverted for SV40 6 months after inoculation with SIV and developed severe bilateral tubulointerstitial nephritis. In contrast, progressive multifocal leukoencephalopathy (PML) occurred in two adult monkeys that were seropositive for SV40 before SIV inoculation, as well as a third adult that was naturally infected with SIV and seropositive for SV40 5 years before death. Large intranuclear inclusions containing abundant polyomavirus particles were limited to either renal tubular epithelial cells or oligodendrocytes. In situ DNA hybridization for SV40 large T antigen further demonstrated that SV40 nucleic acid was localized to either kidney or brain tissue. By immunohistochemical analysis, areas of central nervous system inflammation and demyelination were shown to contain CD68+ macrophages (gitter cells), aggregates of CD8+ T lymphocytes, and numerous gemistocytic astrocytes that labeled for glial fibrillary acidic protein. These observations indicate that rhesus monkeys with SIV-induced AIDS are predisposed to polyomaviral disease, in which SV40 nucleic acid is observed in renal tissue in primary infections and brain tissue after viral reactivation. Furthermore, this organ-specific replication suggests that tissue-tropic strains of SV40 may develop in immunodeficient monkeys.


Differentiation of bursal secretory-dendritic cells studied with anti-vimentin monoclonal antibody. Embryonic and posthatched differentiation of bursal secretory dendritic cells, which express vimentin intermediate filaments, were studied with anti-vimentin (clone 3B4) and anti-cytokeratin (clone Lu5) monoclonal antibodies. Anti-cytokeratin staining revealed that medullary reticular epithelial cells formed a continuous network at every age, whereas the vimentin positive cells were single and showed dendritic appearance. On the basis of location, number, shape, polarized appearance, and Ia staining, the vimentin-positive cells and secretory dendritic cells appeared to be the same cell. Secretory dendritic cell precursors entered the bursal epithelium between 11 and 13 days of embryogenesis. The first vimentin positive cell appeared in the bud of 14-day embryos. Bud formation preceded the appearance of vimentin-positive cells. These observations suggested that the secretory dendritic cell precursor did not express vimentin when it entered the epithelium. Between 15 days of embryogenesis and 2 weeks of posthatch development, the changes in vimentin staining pattern revealed a cytological differentiation of the vimentin-positive cell. During rapid bursal growth, the number of secretory dendritic cells (vimentin-positive cells) increased about 18 times possibly by proliferation of vimentin-negative precursors in the epithelial arches of the corticomedullary border.


Histochemical study of the heart of the axolotl (Ambystoma mexicanum). We have investigated the presence of cells containing monoamines, substance P, and neuron-specific enolase (NSE) in the heart and in the pericardial wall of a urodele amphibian, the axolotl. Fibers containing substance P-like immunoreactivity were present in the heart but not in the pericardial wall. Also present in the heart were small branched cells, which stained metachromatically with toluidine blue. Similar cells were found in the peritoneum and were tentatively identified as mast cells. NSE-immunoreactive fibers were found both in the heart and in the pericardial wall. Small intensely fluorescent (SIF) cells of the pericardial wall contained a high concentration of norepinephrine but no other monoamines, substance P, or NSE. Comparison with data available for the mudpuppy, Necturus maculosus, a closely related amphibian species, suggests that the innervation of the heart in the axolotl is substantially different.


In vitro effects of substance P and somatostatin on lymphoproliferation in rainbow trout (Salmo gairdneri). Lymphocytes from peripheral blood of rainbow trout are put in the presence of increasing concentrations of substance P (SP) and somatostatin (SOM). We have shown that SP stimulates and SOM inhibits lymphoproliferation and that the effects are dose dependent. These results suggest that SP and SOM receptors may exist on fish peripheral blood lymphocytes. When cells are stimulated by PHA or LPS, the presence of SP enhances the response to PHA whereas it only modifies the response to LPS to a slight extent. The presence of SOM inhibits PHA- or LPS-induced stimulation. The inhibition of the proliferation is higher in the case of LPS-stimulated cells. These results suggest that there is an unequal distribution of neuropeptide receptors among the various lymphocyte subpopulations.


Ion channel activity and transmembrane signaling in lymphocytes. Transmembrane signalling refers to the process of transfer of information from the extracellular world into the intracellular space. The information is transduced through several possible pathways. The significance of cell surface dynamics, ion channel activity and drug effects are discussed in the signal transmission, with special reference to Na+ channels and the Ca2+ sensitive potassium channels.


Management of accidental kerosene ingestion. Accidental kerosene ingestion is still a common problem in Libya. It causes considerable morbidity and occasionally mortality. The role and choice of antibacterial agents in its management remain unsettled. Pulmonary damage has been reported as resulting from aspiration. In aspiration pneumonia, anaerobic organisms may be important pathogens and metronidazole may have a place in therapy. The present randomized trial in 100 children with accidental kerosene ingestion assesses the role of ampicillin, carbenicillin and metronidazole in its management. The results are not conclusive but chemoprophylaxis appears to decrease morbidity. Of the various regimens used, the ampicillin/metronidazole combination was found to be slightly better than the others. Further study is recommended.


Maternal and environmental factors associated with infections and undernutrition in young Australian aboriginal children. Forty-eight Aboriginal infants in remote communities in north-west Australia were studied monthly from birth to 2 years. Birthweight and growth were positively associated with maternal health during pregnancy, regular antenatal supervision and lack of drinking alcohol or smoking tobacco as well as with personal and family hygiene. Impaired growth was associated with families which had more expensive consumer goods such as televisions, air conditioners, video cassette recorders and cassette tape players. Most children had impaired growth or frank failure to thrive which was associated with high rates of infections, particularly of the respiratory and gastro-intestinal tracts, and high rates of hospitalization.


Intussusception in children under 2 years of age in the State of Qatar : analysis of 67 cases. Intussusception is one of the leading causes of bowel obstruction in early infancy and childhood. From 1984-1989, 67 patients under 2 years of age with intussusception were diagnosed and treated in our institution. There were 48 boys and 19 girls ranging in age from 2 months to 2 years with a mean of 7.4 months. Presenting symptoms and signs included abdominal pain (96%), vomiting (93%), rectal bleeding (60%) and a palpable mass (67%). Symptoms and signs were present for less than 24 hours in about 80% of cases. Most of the intussusceptions were of the ileocolic type (75%). The overall success rate of hydrostatic barium enema reduction was 49%. The highest rate of reduction by enema was among patients between 9 and 16 months of age (83%). The success rate of barium enema reduction was negligible after 24 hours of cardinal symptoms. Five children underwent surgical exploration without contrast studies because of delayed presentation and signs of an acute abdomen. A pathological lead point was found in only four cases, the commonest being Meckel's diverticulum. The average length of hospitalization was 2.57 days after barium enema reduction and 7.55 days after surgical reduction. There were no deaths. There was no case of perforation during enema reduction. Three children had recurrence within 3 months of initial presentation. The best outcome is associated with early diagnosis and barium enema reduction, or selected surgical intervention when indicated.


Irrational speech occurring in the course of anti-tuberculous therapy--possibly caused by isoniazid. Two girls, both aged 11 years, one with pulmonary tuberculosis and the other with abdominal tuberculosis, were admitted to the Wesley Guild Hospital, Ilesha, Nigeria during the months of June and October, respectively, in 1989. During the course of treatment with combination chemotherapy, both developed irrational speech which responded to temporary withdrawal of isoniazid from the combination. Subsequent progress was uneventful.


Plasma zinc, copper, selenium, ferritin and whole blood manganese concentrations in children with kwashiorkor in the acute stage and during refeeding. Plasma zinc, copper, selenium, ferritin and whole blood manganese concentrations were measured in 22 children with kwashiorkor on admission to hospital and on days 5, 10 and 30 of refeeding. Twenty similarly aged, healthy, well nourished children served as controls. The mean (SEM) zinc, copper and selenium concentrations of 7.5 (0.93), 10.8 (0.64) and 0.29 (0.02) mumol/l, respectively, in the children with kwashiorkor on admission were all significantly lower than the values of 13.7 (0.66), 25.6 (1.72) and 0.72 (0.04) mumol/l in the controls. In contrast, the erythrocyte manganese level of 1.67 (0.09) micrograms/gHb and the median ferritin concentration of 293 micrograms/dl were significantly higher than in the controls. After 30 days there was full clinical recovery with significant weight gain and a return of the plasma albumin, caeruloplasmin, copper and ferritin to normal. However, manganese remained elevated and zinc and selenium concentrations remained significantly low. Our results suggest that nutritional rehabilitation of children with kwashiorkor is incomplete by 30 days and cannot be judged purely by a return of the plasma proteins to normal. Addition of selected trace elements to the diet may hasten full recovery.


The prevention of Indian childhood cirrhosis. Previous studies have led to the hypothesis that the gross hepatic copper storage characteristic of Indian childhood cirrhosis (ICC) is due to the early introduction of animal milk feeds which have been contaminated with copper from brass household utensils. Amongst the families of 100 cases of ICC, the incidence of ICC in children born after dietary advice had been given (1/86) was significantly lower than in older siblings (12/125). This study attempted to document the incidence of ICC and the usage of brass before and after an intervention programme in Pune District advising against this pattern of infant feeding. The study encountered numerous difficulties in data gathering, but documented a fall in ICC prevalence resulting in its virtual disappearance in Pune District. This contrasted with an unchanged incidence in Chandigarh. Although a fall in brass usage was seen in Pune District, this was actually a spontaneous sociological change rather than a result of health education.


Poisoning from henna dye and para-phenylenediamine mixtures in children in Khartoum. Poisoning by a mixture of henna dye and para-phenylenediamine dyes led to the hospitalization of 31 Sudanese children between 1984 and 1989. There was a characteristic clinical presentation. All children presented with an acute and severe angioneurotic oedema and 15 of the cases required emergency tracheostomy for respiratory obstruction. Acute renal failure occurred in five children who recovered after peritoneal dialysis. Mortality was high, all 13 deaths occurring within 24 hours of presentation. Hypotensive shock gave a poor prognosis. It is possible that similar cases may be occurring unrecognized where henna is traditionally used. A programme of public education and restriction of para-phenylenediamine is urgently required in The Sudan and other affected nations. Ingestion was accidental in 12 children, deliberate in 10 and homicidal in three cases. Cutaneous absorption was likely in the remaining six.


The use of herbal remedies in Jamaica.

The use of herbal remedies in Jamaica. A survey of paediatric inpatients at the Tropical Metabolism Research Unit in the University Hospital, Kingston, Jamaica demonstrates that 71% had been treated with herbal remedies before their presentation to the medical services. The risks of the high prevalence of such medication in children are outlined, and two remedies in particular are highlighted because of their potential toxicity.


Hepatotoxicity of high dose salicylate therapy in acute rheumatic fever. Liver function tests, including serum alanine aminotransferase (ALT) activity, serum bilirubin, alkaline phosphatase, serum proteins, blood ammonia levels and intravenous glucose utilization, were monitored in 50 children with acute rheumatic fever receiving anti-rheumatic doses of aspirin. There was a significant increase in blood ammonia levels and serum ALT after aspirin therapy. A significant fall in glucose utilization coefficient was also recorded. Serum alkaline phosphatase, bilirubin and total proteins did not change significantly. Twenty-two of the 50 children recorded a rise in serum ALT; in 12, the rise was five- to tenfold. These 12 children developed adverse symptoms to aspirin. Also, all had a marked rise in blood ammonia levels. The children improved clinically and biochemically on withdrawal of aspirin. There was no constant relationship between hepatocellular function and serum salicylate levels.


Limited impact of a targeted food supplementation programme in Bangladeshi urban slum children. An energy-dense supplementary food, together with nutrition education, was given to a group of moderately malnourished children aged 6-12 months in a poor slum community of urban Bangladesh. An age- and sex-matched control group received only nutrition education. Both groups were followed monthly with respect to weight gain and morbidity. The purpose of the study was to assess the differential impact of a targeted supplementary feeding programme with nutrition education and a nutrition education programme alone on monthly weight gain during 6 months. During the 1st 3 months of the intervention, the monthly weight gain of the supplemented children was 205 g vs 159 g in the control children (p less than 0.05). In the following 3 months, differences in weight gain were no more significant. Several possible explanations for this transient impact are discussed. It is suggested that nutrition education in the control group may have been responsible for the limited difference between the two groups, but seasonal and epidemiological factors may also have played a part.


Efficacy of a therapeutic feeding centre evaluated during hospitalization and a follow-up period, Tahoua, Niger, 1987-1988. Between 1 February 1987 and 31 May 1988 an evaluation of a nutritional rehabilitation centre in Tahoua, Niger was conducted. Among the 381 children admitted to the centre, 61 (16%) had kwashiorkor and 347 (91.3%) were aged between 6 and 29 months. Recovery and death rates were 46.2% and 14.4%, respectively. The median duration of stay until recovery was 21 days. Sixty-two per cent of deaths occurred during the 1st week of hospitalization. Three risk factors for death were identified by the study: patients with kwashiorkor with a weight/height (W/H) less than -3 SD, those with marasmus with a W/H less than -5 SD, and those dehydrated with marasmus. Among children included in the follow-up study after leaving the centre, the risk of dying during the follow-up period among children who absconded was 7.1 times higher than the risk observed among children who recovered. Among the children who recovered, no relapse was observed 3-18 months after they left the centre. This investigation indicates the importance of intensive therapeutic feeding centres in areas with a high prevalence of malnutrition.


Physical activity and anthropometric and functional characteristics of mildly malnourished Senegalese children. This study examines the effects of chronic malnutrition on the functional capacities and physical activity patterns of a group of 100 healthy Senegalese children between the ages of 10 and 13 years. Anthropometric measurements, a sub-maximal step test, spirometric tests and testing of four motor skills (foot races, jumping, throwing, gripping) were conducted and their physical activity was monitored by recording of heart rate every minute for 6 hours. The weights of two-thirds of these children fell below -1 SD from the WHO/NCHS* norm for their ages, their test results were inferior to those of Western children, and the level of their physical activity appeared also to be low. When these children are divided on the basis of weight deficits for age into well nourished and malnourished groups, malnourished children register poorer functional performances than well nourished children, but no difference exists with respect to the intensity of physical activity. These results highlight the negative effect of malnutrition on children's physical performance. The consequences are disturbing as the subsistence in the Sahelian Region depends on substantial physical labour.


Vitamin A status of young Gambian children: biochemical evaluation and conjunctival impression cytology. A pilot study was conducted to examine the vitamin A status of Gambian pre-school children by conventional biochemical means and to evaluate the use of conjunctival impression cytology (CIC) in the detection of subclinical vitamin A deficiency. Children were examined on three occasions to coincide with periods of low and high carotene intakes. Plasma retinol and beta-carotene showed highly significant seasonal changes that reflected the seasonal fluctuations in dietary vitamin A. A high prevalence of low plasma retinol levels suggested that vitamin A deficiency may be a public health problem in this community. Underlying infection and vitamin A instability may have contributed to the low plasma retinol levels and given an overestimate of the prevalence of vitamin A deficiency. Seasonal changes in conjunctival impression cytology were not significant. Abnormal CIC results showed no clear association with plasma beta-carotene or retinol.


Concentration of zinc, copper and metallocalorie ratio in bottle-milks prepared by poor urban families. The concentration of zinc and copper and metallocalorie ratios were measured in samples of bottle-milks fed to infants from a poor urban settlement in Brasilia, Brazil. The bottle-milks fed to infants under 1 year of age from a sample of 40 families were analysed for zinc and copper. Zinc and copper concentrations ranged from 0.7 to 11.5 mg/l (mean 3.37) and from 0.09 to 1.47 mg/l (mean 0.43), respectively. The mean metallocalorie ratio was 1.26 mg/MJ (5.29 mg/1000 kcal) for zinc and 0.17 mg/MJ (0.73 mg/1000 kcal) for copper. Based on minimum recommendations for formulae, 55% of the bottle-milks had both zinc and copper concentrations below 3.2 and 0.4 mg/l, respectively. However, owing to added sugar and high caloric concentrations in the milk preparations, metal:calorie ratios were below the minimum recommendations in 72.5 and 62.5% of cases for zinc and copper, respectively. The variation in zinc and copper concentrations in the milks prepared by mothers/infant caretakers under unsupervized home conditions is wide. Owing to the low availability of zinc in cow's milk preparations, it is of concern that more than half the bottle preparations had zinc and copper concentrations/ratios below the minimum recommendations.


Evaluation of urinary tract infection in malnourished black children. Urinary tract infection (UTI) is a well recognized complication in malnourished children. The need to investigate these patients for underlying renal pathology has not been clearly defined. Seventy-five children with malnutrition were evaluated for UTI by culture of urine obtained suprapubically prior to antibiotic therapy. All patients with UTI were investigated with renal ultrasonography, intravenous pyelography (IVP) and voiding cystourethrography (VCU). Haemoglobin, white cell count, serum urea, creatinine and electrolytes were determined in all the children. The mean age of the children was 15.5 months (range 3-60 months). UTI was diagnosed in 26 (34.7%), of whom 21 (81%) were boys. The overall prevalence of UTI in those with kwashiorkor/marasmic kwashiorkor was 42%. Escherichia coli was the organism most commonly cultured (84.6%). Renal sonography, IVP and VCU were normal in all infected cases and vesicoureteric reflux was not detected in any. This study confirms the high prevalence of UTI in malnourished children. As no anatomical abnormalities were demonstrated in the patients with UTI, imaging of the renal tract other than real sonography does not appear to be indicated in the malnourished child in a first episode of UTI with normal renal function.


A study on delayed hypersensitivity to rotavirus in infancy and childhood. The T-cell responsiveness to rotavirus antigen and phytohaemagglutinin (PHA) together with T-cell total and subsets quantitation was carried out in 50 non-diarrhoeal and six rotavirus diarrhoeal subjects. All individuals in the non-diarrhoeal group responded well to PHA and had normal values for T-cell subsets. The number of positive responders to the rotavirus antigen increased gradually from 0% in the newborns to 92% in older children. The increasing risk of exposure to rotavirus infection is thought to be a chief cause of this age-related variation. All the rotavirus diarrhoeal patients responded well to the rotavirus antigen, indicating a potent test system. The T-cell responses to PHA and the T-cell subsets were significantly low. This could be due to temporary T-cell suppression that may accompany viral infection. Our results are discussed in the context of previous studies.


The prognosis of medical coma in Ibadan: results of multivariate analysis. During a recent prospective investigation of 116 children with medical coma in Ibadan, 52 (45%) survived intact, 23 (20%) developed residual neurological deficits and the remaining 41 (35%) died. This series is concerned with the use of multivariate analysis in predicting outcome of medical coma in an individual patient, based mostly on clinical information (27 variables) obtained within 12 hours of admission. Stepwise logistic regression analysis revealed that only 17 of the 27 variables have independent significance in predicting outcome. When discriminant analysis was performed on these 17 variables so as to obtain a classification function of outcome (survived intact, survived with deficits and dead), 92% of the cases were correctly classified. Eight children were, however, seriously misclassified. These were four intact survivors and four dead cases who were classified as dead and survived without neurological sequelae, respectively. These results suggest that multivariate analysis of clinical and laboratory information obtained in the comatose state can provide predictive information.


Subacute sclerosing panencephalitis in Riyadh, Saudi Arabia. The clinical features and outcome of disease in 14 cases of subacute sclerosing panencephalitis (SSPE) diagnosed at the King Khalid University Hospital, Riyadh during an 8-year period are similar to those described elsewhere. Therapy was associated with arrest of deterioration for 2.5 years in one patient, and with survival after diagnosis for 2-7 years in four others. Many of the cases had initial misdiagnoses because of the frequently bizarre modes of presentation. It is thought that many more cases of SSPE occur in Saudi Arabia and also in many other tropical countries than are currently recognized. The establishment of national SSPE registries is advocated to improve early identification and management of cases.


A study of core domains, and the core domain-domain interaction of cytochrome c fragment complex. To gain insight into the folding mechanism of the cytochrome c complex, we prepared a complete set of homologous and hybrid two-fragment ferric complexes of four different types and related complexes from horse, tuna, yeast iso-l, and Candida cytochromes c. The complexes were characterized for structural properties. Apparent equilibrium constants of the complexes were determined to calculate delta G0 for binding. The results have allowed us to assign four core domains of the complex. A core domain is a structural region containing a hydrophobic core and the surrounding shell which folds and unfolds as a unit. Core domain 1 folds by itself and consists essentially of the right channel structure, found by R. E. Dickerson and colleagues, and a part of the heme. Core domains 2, 3, and 4, respectively, are assigned based on the cores located on the left (the Fe-S bond) and right sides and at the bottom of heme. Evidence of the core domain-domain interaction to stabilize the Fe-S bond, combined with the kinetic studies by G. R. Parr and H. Taniuchi, has led to a model of two alternative folding orders of the core domains for the horse type I complex: domain 1----3----2----4 or 1----2----3----4. Furthermore, delta G0 variation between the complexes has shown non-additive behavior, indicating the existence of a residue-residue interaction between the heme- and apofragments in the complex. Evidence suggests that this interaction in most cases occurs within or through the core groups of the ordered interface between the heme- and the apo-fragments formed by folding of core domains 1, 2, and 3. Evidence also suggests that such core group interaction manifests itself in the interaction to stabilize the Fe-S bond and may be manifested in the core domain-domain interaction.


The pyruvate dehydrogenase complex from the parasitic nematode Ascaris suum: novel subunit composition and domain structure of the dihydrolipoyl transacetylase component. The pyruvate dehydrogenase complex (PDC) from muscle of the adult parasitic nematode Ascaris suum plays a unique role in its anaerobic mitochondrial metabolism. Resolution of the intact complex in high salt dissociates the pyruvate dehydrogenase subunit but leaves the dihydrolipoyl dehydrogenase subunit (E3) and two other proteins with apparent M(r)s of 45 and 43 kDa bound to the dihydrolipoyl transacetylase (E2) core. These proteins are not observable on Coomassie brilliant blue-stained gels of other eukaryotic PDCs, but the 45-kDa protein is similar in apparent M(r), pI, and sensitivity to trypsin to the Kb subunit of the bovine kidney PDH alpha kinase. Acetylation of the ascarid PDC with [2-14C]pyruvate under conditions designed to maximize the incorporation of label into protein yielded only a single radiolabeled subunit, E2. These results confirm earlier reports that the ascarid PDC lacks protein X, an integral component recently identified in other eukaryotic PDCs. About 1.6 to 1.8 mol of 14C was incorporated/mole of E2, suggesting that the ascarid E2 contained two lipoly-bearing domains. Domain mapping of the 14C-acetylated ascarid E2 by limited tryptic digestion identified two lipoyl-bearing fragments with apparent M(r)s of 50 and 34 kDa and two core fragments with apparent M(r)s of 46 and 30 kDa. The ascarid E2 domain structure appears to be similar to that of other E2s. However, it appears that the subunit-binding domain (E2B) of the ascarid E2 may be significantly larger or be flanked by larger than normal interdomain regions. An enlarged E2B domain may be necessary to accommodate the additional binding of E3 to the E2 subunit in the ascarid complex, in the absence of protein X.


The differential diagnosis of primary peritoneal papillary tumors. Primary tumors of the peritoneum are rare. Histological differentiation between papillary mesotheliomas, primary ovarian tumors, borderline tumors of the ovary with peritoneal deposits and primary peritoneal carcinoma may be difficult. The expression of vimentin, keratin, pankeratin, CEA, CA125, CA19-9, S100, B72.3 and BerEP4 was therefore investigated in twelve women with primary malignant peritoneal tumors, twelve women with pleural mesothelioma, eight women with serous ovarian carcinoma and four men with peritoneal mesothelioma. The marker pattern we used was no help in differentiating between metastatic ovarian carcinoma and primary peritoneal carcinomatosis. A combination of the markers S100, B72.3 and BerEP4 helped the distinction between mesotheliomas and the other malignancies. If two or all three markers are detectable, primary peritoneal carcinomatosis or metastatic ovarian carcinoma is the possible diagnosis. If none of the three markers are found, a diagnosis of mesothelioma is highly probable.


Antigenic variation of the bovine ephemeral fever virus glycoprotein. Glycoprotein-specific monoclonal antibodies (MAbs) were used to select escape mutants of bovine ephemeral fever (BEF) virus to determine the escape frequency for different epitopes and to construct an epitope map. At least six antigenic sites were detected by this method and escape frequencies between 10(-2) and 10(-8) were recorded. One new non-conformational site was defined by a MAb, 5A5, which neutralized Berrimah and Kimberley viruses as well as three BEF virus strains. Batch to batch variation was detected in the BB7721 strain of BEF virus when tested for MAb neutralization. Eighteen strains of BEF virus, isolated from blood and insects from a variety of locations in Australia over a period of 33 years, were examined using MAbs and at least one epitope could not be detected in strains isolated since 1975. Implications for vaccine development are discussed.


[Refinement of the spatial structure of the gramicidin A ion channel]. The spatial structure of the gramicidin A (GA) transmembrane ion-channel was refined on the base of cross-peak volumes measured in NOESY spectra (mixing time tau m = 100 and 200 ms). The refinement methods included the comparison of experimental cross-peak volumes with those calculated for low-energy GA conformations, dynamic averaging of the low-energy conformation set and restrained energy minimization. Accuracy of the spatial structure determination was estimated by the penalty function Fr defined as a root mean square deviation of interproton distances corresponding to the calculated and experimental cross-peak volumes. As the initial conformation we used the right-handed pi 6,3 LD pi 6,3 LD helix established on the base of NMR data regardless of the cross-peak volumes. The conformation is in a good agreement with NOE cross-peak volumes (Fr 0.2 to 0.5 A depending on NOESY spectrum). For a number of NOEs formed by the side chain protons, distances errors were found as much as 0.5-2.0 A. Restrained energy minimization procedure had little further success. However some of these errors were eliminated by the change in torsional angle chi 2 of D-Leu12 and dynamic averaging of the Val7 side chain conformations. Apparently, majority of deviations of the calculated and experimental cross-peak volumes are due to the intramolecular mobility of GA and cannot be eliminated within the framework of rigid globule model. In summary the spatial structure of GA ion-channel can be thought as a set of low-energy conformations, differing by the side chain torsion angles chi 1 Val7 and chi 2 D-Leu4 and D-Leu10 and the orientation of the C-terminal ethanolamine group. Root mean square differences between the atomic coordinates of conformations are in the range of 0.3-0.8 A.


The influence of signs prompting motorists to yield before marked crosswalks on motor vehicle-pedestrian conflicts at crosswalks with flashing amber. The purpose of this experiment was to evaluate the effects of signs reading "STOP HERE FOR PEDESTRIANS" alone and in conjunction with advance stop lines on pedestrian safety at multilane crosswalks with pedestrian-activated amber flashing lights. Motorist and pedestrian behaviors measured throughout this experiment included the occurrence of various types of motor vehicle-pedestrian conflicts; the distance that motorists stopped before the crosswalk when yielding to pedestrians; and the percentage of motorists yielding to pedestrians. The introduction of the sign alone 50 feet (15.15 m) before the crosswalk increased the distance before the crosswalk that motorists yielded to pedestrians and reduced the percentage of motor vehicle-pedestrian conflicts whether the flashing light was activated or not. The addition of the advance stop line was associated with further improvements in both measures.


Insulin-like growth factor I and erythropoiesis. The bone marrow, the primary site of hematopoiesis, is a self-renewing system consisting of a unique micro-environment that promotes the differentiation and proliferation of the various hematopoietic cell lines. While many critical factors necessary for red cell production have been identified, the regulation of erythropoiesis has not been completely elucidated. In addition to multi-lineage growth factors (e.g. interleukin 3 or 4) and lineage-specific hematopoietic growth factors (e.g. erythropoietin), several lines of evidence suggest a key role for insulin-like growth factor I (IGF-I). First, growth hormone stimulates erythropoiesis and IGF-I is known to mediate many of growth hormone's actions (somatomedin hypothesis). Second, factors in bovine serum and in serum from an anephric human with erythropoietic activity distinct from erythropoietin have been identified as IGFs. Third, IGF receptors are found on both erythrocyte precursors as well as mature erythrocytes. Fourth, in vitro IGF-I stimulates erythropoiesis in bone marrow cultures. Fifth, IGF-I administration to neonatal or hypophysectomized animals results in increased erythropoiesis in vivo. Recent studies indicate that IGF-I at physiologic concentrations stimulates erythropoiesis and that growth hormone's action is indirect, occurring via IGF-I. The physiologic source of IGF-I for the bone marrow may be delivery from the serum (an endocrine mechanism) or synthesis within the bone marrow by stromal or other cells (a paracrine mechanism). Our recent studies have shown that mouse bone marrow stromal cells secrete both IGF-I and IGF binding proteins (IGFBPs). The role of IGFBPs in erythropoiesis is not known, but they might modulate the local concentration of IGF-I.(ABSTRACT TRUNCATED AT 250 WORDS)


Variable gene usage, physiology and development of Ly-1+ (CD5+) B cells. Two related features of Ly-1+ B cells (CD5+ B cells in man) are their biased specificities and variable gene usage, and their relationship to the bulk of Ly-1- ('conventional') B cells. An emerging model views Ly-1+ B cells as a memory population in the adult animal, whose generation is largely limited to fetal and neonatal development. Selection based on germline-encoded self-reactivity then shapes the adult repertoire.


Biochemical analysis of MHC and non-MHC lymphocyte membrane antigens. Basic biochemical techniques have been skillfully used to characterize MHC-bound peptides and to examine the changes that occur in MHC molecules and lymphocyte membrane receptor complexes upon ligand binding. The fine tuning of these experimental approaches was possible because of the availability of a variety of biochemical and biological reagents.


Polymerase chain reaction for detection of cytokine gene expression. A powerful method to amplify reverse-transcribed RNA, the polymerase chain reaction can be used to measure cytokine gene transcription in a small number of cells, or in cases where there is low mRNA copy number. This technique may be used to obtain qualitative or quantitative determinations of cytokine gene expression. In this review we discuss the various strategies recently described for the evaluation of cytokine expression using the polymerase chain reaction.


Multienzyme membranes for biosensors.

Multienzyme membranes for biosensors. Artificial multienzyme complexes were prepared in which enzymes were covalently bound to polysaccharide structures activated with urea and formaldehyde. Double enzyme complexes of glucose oxidase and catalase, a glucose oxidase and invertase, were prepared by immobilization on to cellulose fabric. Also, catalase was covalently bound to soluble dextran. The resulting multienzyme systems were highly active and stable, making them suitable for use in measuring the concentrations of glucose and saccharose in solutions. The measurements were performed using an amperometric oxygen electrode and multienzyme membranes containing glucose oxidase and catalase for the first substrate, as well as glucose oxidase bound to cheese-cloth and a 'liquid' membrane of dextran-bound catalase. To determine the concentration of saccharose, a multienzyme membrane with bound glucose oxidase and invertase was used in combination with a 'liquid' dextran-catalase. The enzyme electrodes exhibited a measuring range of 0.1-5 mol dm-3 and a response time of 2-3 min. The electrodes may be used for measuring saccharose and glucose concentrations both in fermentation broths and food products.


Organization of galanin-like immunoreactive neuronal systems in weakly electric fish (Apteronotus leptorhynchus). Immunohistochemical procedures were used to investigate the distribution of galanin-like immunoreactive neuronal somata, fiber pathways and apparent termination fields in the gymnotiform brain. Immunoreactive somata were observed only in the hypothalamus and were confined to preoptic, lateral and caudal hypothalamic regions. Within these areas, positive cells tended to be most concentrated in juxtaventricular nuclei. Dense immunoreactive fiber systems originating from hypothalamic regions were seen to project in separate or coalescing fiber bundles to the basal telencephalan, thalamus/tuberal diencephalon, midbrain and brainstem. The density of positive axons and boutons was quite variable, but regions which displayed the most massive network of axons included structures within the hypothalamus itself (anterior periventricular preoptic nucleus, caudal and lateral hypothalamus), ventral telencephalon (superior and ventral subdivisions), thalamic/tuberal areas (central posterior nucleus and tuberal neuropil within the ventral territory of the prepacemaker nucleus) and brainstem nuclei (dorsal reticular nucleus and the medial paralemniscal nucleus). Within these areas axons appeared more randomly distributed and varicose than along fiber tracs, and in counterstained sections were occasionally seen in apposition to unstained neuronal cell bodies and dendrites. In addition, a system of fibers was seen in the neurointermediate lobe of the pituitary. It is concluded that galanin-like immunoreactive neurons in the gymnotiform brain have a more restricted distribution than those in mammals, and that the major fiber systems emanating from the hypothalamus resemble the diverse projections of the tuberomammillary nucleus of higher vertebrates. The anatomy of galanin-like immunoreactive systems in the apteronotid brain suggests a role in neuroendocrine regulation and an involvement with anatomical areas controlling aggressive and courtship behaviour.


Localization of endoderm-specific mRNAs in differentiating F9 embryoid bodies. Primitive endoderm in the peri-implantation mouse embryo differentiates into two separate lineages, visceral and parietal endoderm (VE and PE). F9 teratocarcinoma cells, when grown in suspension in the presence of retinoic acid (RA), differentiate into embryoid bodies (EBs) which can be used as a model system to study the spatially appropriate induction of VE- and PE-specific gene expression. We have used a whole mount in situ hybridization technique to follow the localization of VE-specific AFP and PE-specific tPA mRNAs during EB differentiation. We show that the putative endoderm-specific markers are localized to the endoderm in mature EBs, and that AFP mRNA is localized to the apical edge of the VE in RA EBs. Surprisingly, prior to localization of endoderm-specific markers at the periphery of EBs, these genes are expressed at low to moderate levels in all cells of the EB. Our data suggest that the establishment of endoderm in EBs is position dependent and not the result of sorting out of predetermined, randomly distributed cells. Our observations also suggest a two-step process for establishing endoderm-specific gene expression, involving up-regulation of transcripts throughout the EB prior to the restriction of their expression to the outer differentiating cell layer.


Afferent connections of the rat substantia nigra pars lateralis with special reference to peptide-containing neurons of the amygdalo-nigral pathway. The afferent connections of the rat substantia nigra pars lateralis have been studied using the retrograde axonal transport of fluorescent latex microspheres. The most numerous groups of retrogradely labelled nerve cell bodies were observed bilaterally in the parabrachial complex and several hypothalamic nuclei, whereas the parietal neocortex, the fundus striati, the central nucleus of the amygdala and the bed nucleus of the stria terminalis were labelled on the injected side only. The neuronal projections from the central amygdaloid nucleus to the substantia nigra pars lateralis and lateral part of the rostral pars compacta have additionally been confirmed by anterograde tracing using wheat-germ agglutinin coupled to horseradish peroxidase. The presence of some peptides in this pathway was studied by combining the use of the same retrograde tracer with immunofluorescence after intra-amygdaloid injections of colchicine. With this method, we have demonstrated that Met-enkephalin, dynorphin and neurotensin are probably utilized as neurotransmitters or co-transmitters in the neurons of the amygdalo-nigral pathway.


Testicular and adrenocortical function in healthy men and in men with benign prostatic hyperplasia. The influence of aging upon serum concentrations of testicular steroids, sex hormone binding globulin (SHBG) and pituitary hormones and on adrenal steroid levels and adrenal steroid response to ACTH was studied in 81 healthy men aged 20-87 years. These endocrine variables were also compared in 43 patients with benign prostatic hyperplasia (BPH), aged 58-89 years and in a subgroup of 41 men, aged 58-87 years, from the above mentioned reference population. The normal endocrine aging was characterized by a rise in SHBG levels, decreasing levels of testicular steroids and non-SHBG-bound testosterone (NST) and increasing gonadotropin levels and decreasing concentrations of total estrone. Adrenal androgen levels decreased in the presence of unchanged levels of cortisol and the adrenal steroid response to ACTH changed by decreasing increments in dehydroepiandrosterone (DHA) and increasing increments in 17 alpha-hydroxyprogesterone (17OHP). With the exception of the alterations in SHBG and adrenal androgens, all these changes were finished before the seventh decade of life. BPH patients had elevated levels of testosterone and NST in the presence of normal SHBG and gonadotropin levels, elevated levels of DHA and DHA sulfate (DHAS) in the presence of normal cortisol levels, a "younger" pattern of adrenal steroid response to ACTH as judged from the increments in DHA and 17OHP, elevated ratios between estrone and 4-androstene-3,17-dione suggesting an increased peripheral aromatization and subnormal prolactin levels. BPH patients may be considered as "endocrinologically younger" than healthy subjects. DHA and especially its proximate metabolite 5-androstene-3 beta, 17 beta-diol exert powerful estrogenic effects on the receptor level. Thus the elevated levels of DHA and DHAS in the BPH patients may create an hyperestrogenic condition in addition to the slight hyperandrogenicity caused by the elevated NST levels. Both endocrine aberrations may play a role in the etiology of BPH, in accordance with the dual sex steroid sensitivity of the periurethral glands.


Carnosine, nerve growth factor receptor and tyrosine hydroxylase expression during the ontogeny of the rat olfactory system. The localizations of carnosine, nerve growth factor (NGF) receptor and tyrosine hydroxylase (TH) were studied in the embryonic and postnatal rat olfactory bulb and epithelium by means of single- and double-immunostaining methods. Tyrosine hydroxylase ontogeny was also evaluated at the mRNA level by in situ hybridization. All these molecules were expressed in the olfactory bulb but with different developmental patterns and cellular localization: carnosine immunoreactivity is seen from embryonic day 17 in primary olfactory neurons scattered in the nasal cavity and in fibres projecting from them to the olfactory bulb. Nerve growth factor-receptor immunoreactivity associated with small glial-like cells is visible in some glomeruli starting from the second day of postnatal life. At postnatal day 10 NGF-receptor immunoreactivity is extended to all glomeruli. Periglomerular neurons expressing TH mRNA and protein are present prenatally and their number sharply increases during the early postnatal development. Double-staining methods show that TH and NGF-receptor immunoreactivity do not overlap in cell bodies and processes. In addition, NGF-receptor immunoreactivity is not colocalized with carnosine. These findings definitely exclude NGF-receptor expression in periglomerular and primary olfactory neurons, suggesting that at least part of NGF-receptor expression in the olfactory bulb is associated with glial cells. In addition, they provide the first immunohistochemical data on carnosine ontogeny and confirm at the mRNA level previous studies on the ontogeny of TH protein.


Regulation of progesterone secretion in human syncytiotrophoblast in culture by human chorionic gonadotropin. In the present investigation we sought to define the specific sites in the pathway of placental progesterone biosynthesis that underlie the action of human chorionic gonadotropin (hCG). When the cells were challenged with dibutryl cAMP (dbcAMP), forskolin or isobutylmethylxanthine, they produced significantly higher amounts of progesterone which in the presence of the hCG antibody was reduced to the level of the control set of cells. Trophoblast cells cultured in serum free medium with 25-hydroxycholesterol (25-OHC) produced increased amounts of progesterone. In the presence of hCG antibody at a concentration which neutralized the secreted hCG, the steroid production was completely blocked, even when the 25-OHC was added to the medium. Also, direct quantitation of the cytochrome P450 SCC enzyme in the absence of hCG indicated a significant decrease. The exogenous addition of low density lipoproteins (LDL) increased the progesterone secretion by the trophoblast cells in culture. Neutralization of hCG by the antibodies, however, drastically reduced the LDL induced progesterone secretion, which was restored by the addition of dbcAMP to the medium. Based on these findings, we suggest a stimulatory effect of hCG on normal trophoblast cells at the level of LDL utilization and cytochrome P450 SCC enzyme. Since dbcAMP could mimic these actions of hCG, the data suggest a possible autocrine/paracrine role of hCG on the trophoblast cells. An additive effect of hCG and cAMP on progesterone secretion observed in our studies, indicate that apart from hCG, adenylate cyclase activity may also be regulated by other factors.


Sanctuary site relapse in chemotherapy-treated testicular cancer. The testis and central nervous system (CNS) may act as sanctuary sites for testicular germ cell tumours, as cytotoxic drugs penetrate these areas less well than systemic sites. We describe three patients who relapsed in the testis (one patient) or CNS (two patients) after receiving chemotherapy for responsive systemic disease. All three were asymptomatic at relapse, which was first manifest by rising tumour marker levels. These sanctuary site relapses were managed locally with surgery +/- radiotherapy. Two patients were rendered disease-free; one died of progression of his local disease only. Sanctuary site tumour should be considered when relapse occurs in the setting of otherwise chemosensitive disease; local therapy may be curative.


A preoperative single course of high-dose cisplatin and bleomycin with glutathione protection in bulky stage IB/II carcinoma of the cervix. One course of preoperative chemotherapy including high-dose cisplatin (40 mg/m2 daily for 5 consecutive days) with glutathione protection and bleomycin (15 mg on days 2, 8 and 9) was administered to 27 patients with bulky operable cervical carcinoma (stage IB/II) in a pilot study. In all patients the tumor diameter was greater than 4 cm. Surgery (radical hysterectomy with pelvic and para-aortic lymphadenectomy) was planned within one month of chemotherapy. In 27 evaluable patients, nausea/vomiting was the most pronounced side effect. Significant (but transient) increases in serum transaminases were detected in 19 patients. Electrolyte imbalance (hypokalemia) was detected in 6 patients (one with hypocalcemia). These reversible effects were not associated with other signs of renal toxicity. Objective clinical responses were observed in 21 patients, 18 of them partial and 3 complete responders (pathologically confirmed in 2). Radical hysterectomy with pelvic and para-aortic lymphadenectomy was performed with no particular complications. The shrinking of bulky tumor made the operation easier, especially in parametrial resections. High-dose cisplatin chemotherapy prior to surgery is feasible with acceptable toxicity. The encouraging results of this study warrant further investigations to define the role of neoadjuvant therapy.


Treatment of high-risk, nonseminomatous testicular cancer with cisplatin, ifosfamide and bleomycin: long-term results. Thirty-four patients with stage IIC (unresectable, retroperitoneal tumor mass (RTM) greater than 5 cm), stage IVC (minimal lung metastases less than 10 cm3 and RTM greater than 5 cm) and IVD (lung metastases greater than 10 cm3 and RTM greater than 5 cm), who had not received previous chemotherapy, were treated with cisplatin (40 mg/m2, on days 2-4), ifosfamide (5 g/m2, on days 1 and 5) and bleomycin (30 mg, on days 1, 8, 15) (PIB), every 21 days. Twenty of the 34 patients (59%) achieved a complete remission (CR). Furthermore, five patients (15%) showed no evidence of disease (NED) after surgical removal of residual tumor masses (NED rate of 74%). A tumor marker-negative partial remission (PR) occurred in 3/34 patients (9%), and a tumor marker-positive PR in another 3/34 patients (9%). Three patients did not respond to this regimen. At a median follow-up period of 38 months (range, 15-47 months), 26/34 patients (76%) were alive, 21 (62%) of them without evidence of disease and three with a stable tumor marker-negative remission. Major toxicity consisted of myelosuppression, neurotoxicity and nephrotoxicity. Chemotherapy-related mortality occurred in two patients (one septicemia and one bleomycin-induced lung fibrosis). In conclusion, PIB is an effective induction regimen in patients with high-risk NSTC. However, controlled clinical trials are necessary to prove the superiority of dose intensification schedules.


Single agent activity of carboplatin in patients with previously untreated non-seminomatous germ cell tumours. The response to a single course of carboplatin has been investigated in 12 patients with previously untreated non-seminomatous testicular germ cell tumours. Patients received one course of carboplatin at a dose calculated to achieve a target area under the free carboplatin plasma concentration versus time curve (AUC) of 7 mg/ml x mins using the formula: dose (mgs) = target AUC x (GFR + 25). Response to carboplatin was assessed after a single course and treatment was then continued on the POMB/ACE schedule. Ten of 12 patients had either a greater than 50% decrease in serum HCG and/or AFP levels or a greater than 50% decrease in tumour volume after a single course of carboplatin. No patient had evidence of disease progression after carboplatin. This study demonstrates that single agent carboplatin is highly active in patients with non-seminomatous testicular germ cell tumours and thus provides evidence to justify its inclusion in chemotherapy combinations.


Palliative care: a Yorkshire approach.

Palliative care: a Yorkshire approach. Palliative care services have traditionally tended to concentrate on the needs of patients with cancer. The philosophy of Yorkshire Health is to extend those services to all people with life-threatening diseases. The progress in service provision which has taken place in the region is described, but work still needs to be done in training professionals to provide the appropriate level of skilled care to patients in hospitals, hospices and community.


Increased thermal stability of proteins in the presence of naturally occurring osmolytes. Organisms and cellular systems which have adapted to stresses such as high temperature, desiccation, and urea-concentrating environments have responded by concentrating particular organic solutes known as osmolytes. These osmolytes are believed to confer protection to enzyme and other macromolecular systems against such denaturing stresses. Differential scanning calorimetric (DSC) experiments were performed on ribonuclease A and hen egg white lysozyme in the presence of varying concentrations of the osmolytes glycine, sarcosine, N,N-dimethylglycine, and betaine. Solutions containing up to several molar concentrations of these solutes were found to result in considerable increases in the thermal unfolding transition temperature (Tm) for these proteins. DSC scans of ribonuclease A in the presence of up to 8.2 M sarcosine resulted in reversible two-state unfolding transitions with Tm increases of up to 22 degrees C and unfolding enthalpy changes which were independent of Tm. On the basis of the thermodynamic parameters observed, 8.2 M sarcosine results in a stabilization free energy increase of 7.2 kcal/mol for ribonuclease A at 65 degrees C. This translates into more than a 45,000-fold increase in stability of the native form of ribonuclease A over that in the absence of sarcosine at this temperature. Catalytic activity measurements in the presence of 4 M sarcosine give kcat and Km values that are largely unchanged from those in the absence of sarcosine. DSC of lysozyme unfolding in the presence of these osmolytes also results in Tm increases of up to 23 degrees C; however, significant irreversibly occurs with this protein.(ABSTRACT TRUNCATED AT 250 WORDS)


Gramicidin channels that have no tryptophan residues. In order to understand how aromatic residues modulate the function of membrane-spanning proteins, we examined the role of the four tryptophans in gramicidin A (gA) in determining the average duration and permeability characteristics of membrane-spanning gramicidin channels; the tryptophan residues were replaced by tyrosine (gramicidin T, gT), tyrosine O-benzyl ether [gramicidin T(Bzl), gT(Bzl)], naphthylalanine (gramicidin N, gN), and phenylalanine (gramicidin M enantiomer, gM-). These analogues form channels with durations and conductances that differ some 10- and 16-fold, respectively. The single-channel conductance was invariably decreased by the Trp----Yyy replacement, and the relative conductance alterations were similar in phosphatidylcholine (DPhPC) and monoglyceride (GMO) bilayers. The duration variations exhibited a more complex pattern, which was quite different in the two membrane environments: in DPhPC bilayers, gN channels have an average duration that is approximately 2-fold longer than that of gA channels; in GMO bilayers, the average duration of gN channels is about one-tenth that of gA channels. The sequence-dependent alterations in channel function do not result from alterations in the channels' peptide backbone structure, because heterodimers can form between the different analogues and gramicidine A, and there is no energetic cost associated with heterodimer formation [cf. Durkin, J. T., Koeppe, R. E., II, & Andersen, O. S. (1990) J. Mol. Biol. 211, 221]. The alterations in permeability properties are consistent with the notion that Trp residues alter the energy profile for ion permeation through long-range electrostatic interactions.


Moderate oxidation of hypertriglyceridemic low-density lipoprotein causes apolipoprotein B epitope change and enhances its uptake by macrophages. We prepared monoclonal antibody (MabB4) that selectively binds to acetylated low-density lipoprotein (LDL). Native hypertriglyceridemic LDL (HT-LDL) obtained from IIb and native normotriglyceridemic LDL (NT-LDL) from type IIa scarcely bound with MabB4. When these LDL were oxidized moderately by incubation with copper ions, the binding of MabB4 to HT-LDL was enhanced compared to that of NT-LDL, although the contents of the hydroperoxide they produced were the same. The incorporation of moderately oxidized HT-LDL into macrophages was enhanced compared to that of NT-LDL, and the rate of incorporation parallel the binding of LDL for MabB4. These results suggested that moderate oxidation of HT-LDL expressed some apolipoprotein B epitope on the surface of acetylated LDL to a much greater degree than NT-LDL, and that this expressed epitope might work as a ligand of moderately oxidized HT-LDL for the recognition by macrophages.


Proliferative responses of a bovine leukemia virus-infected lymphoblastoid B-cell line by its culture supernatant and cytokines. The growth-promoting activity in the culture supernatant of bovine lymphoblastoid B-cell lines (BL2M3 and BL312) were examined. BL2M3 cells proliferated well in response to conditioned medium (CM) obtained from BL2M3 and BL312 cell cultures. These BL2M3 and BL312 CM were used as sources of BL2M3 cell growth-promoting factor (BL2M3-GPF). BL2M3-GPF was sensitive to acid (pH 2) and alkali (pH 10) and was heat-labile. Proliferative responses of BL2M3 cells were not induced by human recombinant (r)IL 1, rIL 2, rIL 6, granulocyte-colony stimulating factor (rG-CSF) or tumor necrosis factor (rTNF)-alpha. Human low molecular weight B cell-growth factor (LMW-BCGF) was, however, capable of augmenting the proliferation of BL2M3 cells. BL2M3 cells formed clusters in response to LMW-BCGF, whereas they showed single and discete appearance in the presence of BL2M3-GPF. These results suggested that bovine lymphoblastoid B-cell lines might release and respond to the growth-promoting factor for in vitro proliferation of its own cell line, BL2M3.


Changes in intracellular and cell surface localization of Le(x) epitope during germ cell differentiation in fetal mice. Changes in the intracellular and cell surface localization of Lewis x (Le(x)) determinants in germ cells during fetal development in mice were examined by light and electron microscopy. Light microscopically, in undifferentiated gonads on day 12 post coitum (p.c.), the anti-Le(x) monoclonal antibody (MAb) was specifically bound to the plasma membranes and to the cytoplasmic granule-like structures of germ cells. In the testes on day 13 p.c., most of the germ cells were enclosed within the testicular cords and showed an MAb-positive reaction which was restricted mainly to the cytoplasmic granule-like structures. The reaction on the plasma membranes almost disappeared. On the other hand, the ectopic germ cells still showed a positive reaction on their plasma membranes. In the ovaries on day 13 p.c., the germ cells also exhibited positive reactions both on the plasma membranes and on the granule-like structures. Immunoelectron microscopic observations agreed well with these light microscopic observations in such a way that both the plasma membranes and the "small dense bodies" (SDB) were positive in undifferentiated gonads on day 12 p.c. In the germ cells organized into the testicular cords, the reaction to anti-Le(x) MAb became restricted to the SDB. These results may indicate that such intracellular changes in Le(x) determinants during germ cell differentiation are associated with the enclosure of germ cells within the testicular cords.


Molecular cloning and immunological analysis of immunodominant piroplasm surface proteins of Theileria sergenti and T. buffeli. cDNA libraries of Theileria sergenti and T. buffeli piroplasms were constructed in lambda gt11 and screened with rabbit anti-piroplasm sera. A major antigen of T. sergenti (33 kDa) and that of T. buffeli (34 kDa) was identified from the recombinant phages by using recombinant antigen-selected monospecific antibodies. The reactivities of the cloned proteins with rabbit antisera, infected calf sera and mouse monoclonal antibody suggested that the 33 and 34 kDa proteins expressed species-common and species-specific epitopes. The DNA probes from these recombinant clones showed species-specific hybridizations in Southern blotting with genomic DNA from piroplasms. These results indicate that the Japanese T. sergenti can be distinguishable from the Australian T. buffeli with regard to a polymorphism of the major immunodominant proteins of piroplasm.


Plasminogen activators in the human endometrium, cellular origin and hormonal regulation. Endometrial tissue explants in culture were found to release urokinase-type plasminogen activator (u-PA) and tissue-type plasminogen activator (t-PA). In order to identify their cellular origin and possible hormonal regulation, enriched cultures of glandular epithelial cells and stromal cells were prepared from fresh endometrium, and the cultures treated with hormones. Both epithelial and stromal cell cultures were found to secrete u-PA and t-PA. Treatment of epithelial cell cultures with oestradiol, progesterone and DH-testosterone had no effect on the secretion of t-PA or u-PA. In stromal cell cultures, on the other hand, the secretion of u-PA was significantly reduced after treatment with progesterone, whereas oestradiol and DH-testosterone had no effect. This reduction of u-PA antigen in the tissue culture medium did not result from a reduction of the relative level of u-PA mRNA in the cells, suggesting that the synthesis of u-PA was not reduced. Alternatively, an increased clearance of u-PA by the cells from the medium may explain the reduction. This in vitro observation probably reflects the in vivo reduction of u-PA in endometrial secretion during the secretory phase.


The interferons and their use in condyloma acuminata. There are three naturally occurring interferons: alfa, beta, and gamma. Alfa, derived from lymphoblastic tissue, is approved by the Food and Drug Administration for the treatment of condyloma acuminata (genital or venereal warts). Genital warts are caused by human papillomaviruses, of which more than 50 subtypes have been described. Traditional therapies have centered on destruction of the lesions by either cytotoxic or physical modalities. Intralesional interferon exerts its antiviral effects on infected cells without causing damage to the surrounding tissue. In general, success rates with intralesional interferon alfa are comparable to traditional modalities. There is also evidence that interferon alfa might be particularly useful in the treatment of lesions that have failed to respond to other modalities.


Meniscus-like synovial fold in the atlantoaxial (C1-C2) joint. Some of the synovial joints in the human body have a fibrocartilaginous disc interposed between the joint surfaces to absorb or evenly distribute loads. Examples of fibrocartilagenous discs include the intervertebral disc, knee joint meniscus, and triangular fibrocartilages in the distal radioulnar joint and the acromioclavicular joints. The joint capsule and the surrounding tissue from nine cervical spines (18 C1-C2 joints) were dissected and prepared for gross examination and histology. We found meniscus-like synovial folds in 13 of the 18 atlantoaxial joints. These folds were located at the anteromedial and posteromedial aspect of the joint. Each synovial fold was of semilunar shape, with a thickened outer edge and thin inner edge giving a wedge-shape cross section. In one case, the synovial fold was grossly similar in appearance to a knee joint meniscus, and on histological examination there was evidence of cartilagenous metaplasia in part of the fold. The findings are compared with the limited data reported in the literature.


Clinical value of measuring the interferon-induced enzyme 2'-5'-oligoadenylate synthetase in children. 2'-5'-oligoadenylate synthetase, an interferon-induced enzyme and a sensitive indicator of the presence of interferon, was measured in peripheral blood mononuclear cells in children with inflammatory disorders of known and unknown origin in order to assess the value of such a test in patient management. Differences in median 2'-5'-oligoadenylate synthetase values in groups of children with viral or bacterial diseases or healthy children were observed. Considerable overlap of the 2'-5'-oligoadenylate synthetase range in the three groups was observed. All children with acute viral infections had increased levels early in their disease and a low value clearly argued against an acute viral infection. However, as more than one-third of children with bacterial diseases displayed elevated 2'-5'-oligoadenylate synthetase concentrations, distinction between a viral and bacterial cause of disease by measuring this enzyme was difficult. Estimation of 2'-5'-oligoadenylate synthetase concentrations in patients with inflammatory diseases of unknown origin, including juvenile rheumatoid arthritis, systemic lupus erythematosus, idiopathic uveitis and glomerulonephritis did not contribute to establishing a diagnosis or measuring disease activity.


Further studies on platelet-mediated neurotoxicity. The mechanism of ischemic neuronal injury is not fully resolved. The present view is that vascular occlusion per se does not fully account for the extent of neurological dysfunction. We hypothesized that platelet secretory products might contribute to ischemic neuronal injury in the central nervous system (CNS) (Joseph et al., Stroke, 20 (1989) 38-44 and 1316-1319). Our preliminary studies using organotypic rat spinal cord cultures exposed to human platelet and its secretory products, revealed that platelet product(s) had neurotoxicity. Further studies, using the same methods, were conducted here, with the addition of several refinements such as use of gel-filtered platelets (as opposed to washed platelets), adding additional relevant controls including platelet membranes, red blood cells and washed rat platelets. The results confirmed our initial finding that an agent(s) in platelet secretion is neurotoxic. Subsequently, we identified serotonin (5HT), a major platelet product, as having toxic effects on neurons. This toxicity of 5HT appeared to be blocked by ketanserin, a 5HT2 receptor antagonist. Judging by the concentrations of 5HT that demonstrated neurotoxicity in these in vitro studies, it appears that products secreted from activated platelets could have pathological significance in vivo.


Microglia isolated from rat brain secrete a urokinase-type plasminogen activator. In a previous study, we found particular proteases which degrade myelin basic protein (MBP) in a conditioned medium of cultured rat brain microglia. The MBP degrading activity in microglial-conditioned medium (Mic-CM) increased markedly in the presence of plasminogen. By Sephadex G-150 column chromatography, plasminogen-dependent MBP degrading activity was eluted at the position of about 47 kDa and 28 kDa. Furthermore slight plasminogen-dependent protease activity in the presence of fibrin (tissue plasminogen activator activity) was detected at a molecular weight of about 68 kDa. The two molecular forms (47 kDa and 28 kDa) of plasminogen-dependent protease were demonstrated by casein-zymography, and it was suggested that they were urokinase type-plasminogen activators (uPA). This suggestion was confirmed by immunoblotting using anti-uPA antiserum. The unique 28 kDa type was considered to be produced from the 47 kDa form by limited proteolysis. Secretion of PA from microglia was demonstrated by cell zymography. In contrast, significant secretion of plasminogen activator inhibitor could not be detected in the Mic-CM. In addition, lipopolysaccharide significantly decreased the secretion of PA from microglia, while interleukin-1 and basic fibroblast growth factor enhanced the secretion.


A 35 kDa Fos-related antigen is co-localized with substance P and dynorphin in striatal neurons. The rat striatum after dopamine denervation followed by repeated apomorphine treatment was examined for the co-expression of c-fos and Fos-related antigens with dynorphin, substance P and [Met5]enkephalin using Western blot and immunohistochemical techniques. Administration of apomorphine, a dopamine agonist, elevated the level of 35 kDa Fos-related antigen which co-localized with dynorphin and substance P, but not enkephalin, in striatal neurons.


Ultrastructural localization of nitric oxide synthase immunoreactivity in guinea-pig enteric neurons. Electron microscopic immunocytochemistry was used to localize immunoreactivity for nitric oxide synthase (NOS) in whole-mount preparations of myenteric plexus and circular muscle from guinea-pig ileum. NOS immunoreactivity was patchily distributed in myenteric neurons and was not specifically associated with any subcellular organelle or with the plasma membrane. This localization leaves unanswered the question of how nitric oxide is stored and released. NOS immunoreactive fibres in the circular muscle were found closer than 100 nm to muscle cells. NOS immunoreactive nerve fibres made synaptic contacts with NOS immunoreactive and non-immunoreactive enteric neurons. These results indicate that nitric oxide may regulate the activity of both myenteric neurons and smooth muscle.


An endothelial ligand for L-selectin is a novel mucin-like molecule. The adhesive interaction between circulating lymphocytes and the high endothelial venules (HEV) of lymph nodes (LN) is mediated by lymphocyte L-selectin, a member of the selectin family of cell adhesion proteins. Previous work has identified a sulfated 50 kd glycoprotein (Sgp50) as an HEV ligand for L-selectin. We now report the purification of this glycoprotein and the utilization of the derived N-terminal amino acid sequence to clone a cDNA. The predicted sequence reveals a novel, mucin-like molecule containing two serine/threonine-rich domains. The mRNA encoding this glycoprotein is preferentially expressed in LN. Antibodies against predicted peptides immunoprecipitate Sgp50 and stain the apical surface of LN HEV. These results thus define a tissue-specific mucin-like endothelial glycoprotein that appears to function as a scaffold that presents carbohydrates to the L-selectin lectin domain.


An antigenic determinant on human centromere protein B (CENP-B) available for production of human-specific anticentromere antibodies in mouse. Centromere protein B (CENP-B) is one of the centromere DNA binding proteins constituting centromere heterochromatin throughout the cell cycles. Some components of mammalian centromeres including CENP-B are target antigens for autoimmune disease patients, often those with scleroderma. Recent isolations of CENP-B genes from human and mouse suggested that CENP-B was highly conserved among mammals. From the previous analysis of the reactivity of patient anticentromere sera, two autoepitopes have been located on the DNA binding domain at the amino-terminal region. The amino acid sequences for both the epitopes are perfectly conserved in the two species, human and mouse. In this study, to identify a human-specific antigenic determinant, the remaining two epitopes were further located in separate carboxyl-terminal regions of human CENP-B. Although the amino acid sequence of one epitope is identical to that of the corresponding region in mouse CENP-B, the other has a less homologous sequence. To confirm that the latter epitope was available for production of human-specific anticentromere antibodies, mice were immunized with the recombinant human CENP-B product. One serum that exclusively stained human centromere structure, but not that of other mammals, was identified in the immunofluorescence microscopic observation. The epitope analysis showed that the less conserved one was recognized by this serum. These results suggested that the corresponding region defines the antigenic determinants for the species specificity.


Isolation and characterization of nuclei from rice embryos. A method has been developed to isolate pure preparations of nuclei in high yield from commercially available viable rice embryos (germ), employing extraction with buffer solution containing glycerol (without detergent) and polyamine, followed by centrifugation on a 30% Percoll cushion. The intactness of the isolated nuclei was confirmed by light microscopy as well as electron microscopy. The protein profiles of both whole nuclei and nuclear extracts obtained by SDS-PAGE, organellar marker enzyme activities, DNA and RNA analyses, and in vitro RNA synthesis, all indicate that the highly purified nuclei are isolated from rice embryos.


Inhibitory effects of bis(2-aminohexyl)disulfide and its analogues on polymorphonuclear leukocyte functions in vitro. Water soluble analogues of the anti-inflammatory compound, bis(2-aminopropyl)disulfide dihydrochloride (compd. I) with a butyl (II), phenyl (III), benzyl (IV) or pyrrolidinyl group (V) instead of the methyl group were synthesized, and their effects on the functions of cells related to inflammation were studied in vitro. Compounds II, III and IV showed much higher inhibitory activity than compd. I on formyl Met-Leu-Phe (FMLP)-induced O2(-)-generation of polymorphonuclear leukocytes (PMNs) and platelet aggregation. Compound II showed the strongest activity among the compounds (IC50 values: 2.6 microM). The inhibition of O2(-)-generation of PMNs by compd. II was the most effective when FMLP was used as a stimulant rather than when phorbol myristate acetate, A-23187 and opsonized zymosan were used. However, compd. II was not an O2(-)-scavenger. Compounds II, III and IV significantly inhibited a series of activation processes in PMNs, chemotaxis, phagocytosis and lysosomal enzyme release at doses ranging from 10 to 100 microM. Under these doses, compds II, III and IV did not affect the histamine release from mast cells or the hemolysis of erythrocytes. These results strongly suggest that the anti-inflammatory action caused by compd. II and its analogues was at least partly due to inhibition of several functions of PMNs and platelets.


Characterization of a mouse hepatocyte growth-stimulating factor in serum of mice treated with carbon tetrachloride. The physicochemical and biological properties of a mouse hepatocyte growth-stimulating factor (mHGSF), whose amount in mouse serum increased markedly 24 h after carbon tetrachloride administration (E. Gohda et al., Life Sci. 46, 1801 (1990)), were examined. This factor was a heat-labile protein with a molecular weight of 75000. Its activity was sensitive to disulfide reduction. Maximal stimulation of deoxyribonucleic acid synthesis in cultured rat hepatocytes by this factor was greater than that by acidic fibroblast growth factor (acidic FGF) or mouse epidermal growth factor (EGF) and was comparable to maximal stimulation by human hepatocyte growth factor (hHGF), a heterodimer with a molecular weight of about 85000. The effect of mHGSF was additive to the maximal effects of acidic FGF and EGF and was synergistic with the maximal effect of insulin, but was neither additive nor synergistic with the maximal effect of hHGF. The mHGSF activity was not inhibited by a neutralizing anti-hHGF antiserum, which recognizes nonreduced hHGF but not reduced heavy and light chains of hHGF. mHGSF did not show any cross-reactivity to anti-hHGF monoclonal and/or polyclonal antibodies as measured by an enzyme-linked immunosorbent assay for hHGF. These results suggest that mHGSF is a hHGF-like factor with some structural difference from hHGF.


Comparison of flanking regions of the 5S ribosomal ribonucleic acid genes in Leptospira biflexa and Leptospira interrogans. One of the genes encoding the 5S ribosomal ribonucleic acid (rRNA) for the Leptospira biflexa strain Patoc I was isolated and sequenced. The physical maps of the 5S rRNA genes in Leptospira were constructed. The strains of Leptospira biflexa had two genes on their chromosome; these two 5S rRNA genes were located several kb apart and sequences flanking these genes were divergent. In contrast to saprophytic leptospires, maps in parasitic leptospires that had only one gene for 5S rRNA on their genome were highly conserved and the physical maps of the genes in almost all strains were similar.


Permeability of rat atrial endocardium, epicardium, and myocardium to large molecules. Stretch-dependent effects. Atrial distension, which stimulates atrial natriuretic peptide secretion by atrial myocytes, also stretches nonmuscle cells. In a noncontracting in vitro preparation of combined right and left atria we demonstrated by electron microscopy that, at 37 degrees C, transition from zero pressure to a physiological distending pressure of 5.1 mm Hg rapidly rendered atrial endocardial endothelium permeable to the macromolecular probes horseradish peroxidase (HRP; M(r), approximately 40,000) and wheat germ agglutinin-HRP (M(r), approximately 70,000); each probe was introduced at the atrial cavitary endocardial surface. Stretch-dependent permeabilization was also demonstrable in spontaneously contracting atria, was reversed by removing the distending pressure, and was unaffected by varying external Ca2+ concentration from 0.2 to 1.4 mM or by experimental perturbations that markedly decrease ANP secretory rates. Although transendocardial HRP and wheat germ agglutinin-HRP passage required stretch, native ferritin (M(r), = 500,000) could traverse unstretched endocardium. Probes were detected in noncoated endocardial vesicles and intercellular junctions between endocardial cells, but the relative contributions of vesicular transcytosis and paracellular diffusion could not be determined. Although HRP entered plasmalemmal caveolae of myocytes in stretched atria, myocytes did not internalize HRP by fluid-phase endocytosis. Distending pressure also caused apparent flow reversal in thebesian blood vessels, with retrograde transfer of HRP across the endocardium into the myocardium. HRP and ferritin presented at the external surface of the epicardium (visceral pericardium) were endocytosed by mesothelial cells, entered junctions between mesothelial cells, and readily crossed the epicardium of both stretched and unstretched preparations.


Tangential neuronal migration in the avian tectum: cell type identification and mapping of regional differences with quail/chick homotopic transplants. This paper is a sequel to a previous report, using quail/chick chimeras with partial tectal transplants, in which a tangential invasion of host (chick) tectal territories by cells originating in the quail graft was demonstrated. The cells displaying this secondary tangential migration appeared restricted to two strata (stratum griseum centrale (SGC) and stratum griseum et fibrosum superficiale (SGFS)). Here we describe the morphology of the tangentially displaced neurons, as well as their overall distribution in the host tectal lobe, by means of an antibody that specifically recognizes quail cells, staining them in a Golgi-like manner. Neurons that migrated into the SGC are identified as multipolar projection neurons, typical of this stratum. The majority of cells that migrated into the SGFS correspond to horizontal neurons, as was also corroborated by observations in Golgi-impregnated material. These horizontal cells are concentrated in laminae b, d and f, where their processes form well delimited axonal plexuses. In confirmation of previous results, SGC neurons have a limited range of migration, whereas SGFS cells translocate across much longer distances. In reconstructions of appropriate cases, a remarkable polarity was noted. Significant invasion of chick tectum by quail cells mostly occurred in the rostral half of the host tectum. The long-range migration of superficial horizontal cells frequently reached, but did not cross, the rostral tectal boundary. Conversely, tangential migration in the caudal half of the host tectum was scarce and coincided with a typical arrangement of quail-derived radial columns interdigited with chick-derived columns. These findings are discussed in relation to existing data on immature neuronal populations, molecular marker distribution and polarity of the avian optic tectum.


Developmental expression of the 3-fucosyl-N-acetyl-lactosamine/CD15 epitope by an olfactory receptor cell subpopulation and in the olfactory bulb of the rat. In the olfactory epithelium a subpopulation of receptor cells were CD15 (3-fucosyl-N-acetyl-lactosamine)-positive at their somata and their processes. Cytomorphology and location implied their mature character. The labeled neurons were associated with olfactory cilia and showed a unique distribution within the olfactory epithelium and a remarkable numerical change during postnatal ontogeny. The axons of the marked olfactory receptor cells possibly play a role in guiding other axons to their appropriate targets in the olfactory bulb. In the olfactory bulb immunoreactivity was predominantly found in the glomerular and in the external plexiform layer and showed an age-related shift from the superficial to the deep layers.


Characteristics of the interactions between acid glycosaminoglycans and proteins in normal human plasma as revealed by the behaviour of the protein-polysaccharide complexes in ultrafiltration and chromatographic procedures. Acid glycosaminoglycans were isolated from normal human plasma: (a) following fractionation of plasma (with protease inhibitors) on Sephadex G-200; (b) by ultrafiltration through membranes with retention of molecules above 50 kDa, with and without previous addition of NaCl, Triton X-100, urea, or guanidine HCl; (c) by filtering on Ecteola-cellulose either untreated plasma or after treatment with NaCl, urea, Triton X-100, papain or NaOH. More than 95% of plasma glycosaminoglycans interact with plasma proteins to give complexes that exhibit reproducible behaviour on Sephadex G-200 and are retained by ultrafiltration membranes, which the 12-20 kDa polysaccharide chains do filter. High charge plasma glycosaminoglycans show ionic interactions with proteins, while low charge glycosaminoglycan interactions are resistant to Ecteola charged groups, to 0.5% Triton and 4 M urea, while not to 4 M guanidine HC1. Some glycosaminoglycan-protein complexes appear resistant to proteolysis, suggesting that they may originate from lymphocytes. The simple method utilized for plasma GAG measurement may represent an useful tool in clinical practice.


Determination of fetal hemoglobin by cation exchange liquid chromatography. A sensitive liquid chromatographic method for the analysis of the major component of fetal hemoglobin (HbF0) in blood using high-resolution strong cation exchange column is described. The imprecision of the method was assessed in 20 repeated assays of blood samples containing 0.67%, 0.97%, 2.2% and 3.3% of HbF0. Within-assay CVs were 5.0%, 2.8%, 1.8% and 1.5% and between-assay CVs 5.8%, 3.7%, 1.9% and 2.7%, respectively. The HbF0 concentration was stable in EDTA blood for at least 20 days at +8 degrees C and 3 days at +22 degrees C. Blood HbF0 levels (mean +/- S.D.) were 0.41 +/- 0.20% for 21 healthy males, and 0.48 +/- 0.19% for 22 healthy non-pregnant females. The method is simple and reproducible, and allows the sensitive determination of blood HbF0.


Serum alpha 1-antitrypsin in patients with hepatocellular carcinoma. To evaluate the diagnostic value of alpha 1-antitrypsin (alpha-AT) as a tumor marker for hepatocellular carcinoma (HCC), we studied the serum levels of alpha-AT by rocket immunoelectrophoresis and alpha-fetoprotein (alpha-FP) by radioimmunoassay in 46 proven HCC patients, 43 cirrhosis patients and 200 healthy blood donors. The mean alpha-AT level of the 46 patients with HCC (4.8 +/- 2.7 mg/ml) was significantly higher than that of 200 healthy control subjects (1.7 +/- 0.7 mg/ml) (P less than 0.0001). The sensitivity of alpha-AT in 24 patients with high level of alpha-FP (greater than 400 ng/ml) and 22 patients with low level of alpha-FP (less than 400 ng/ml) were 96% and 64%, respectively. There was no substantial correlation between alpha-FP and alpha-AT in the two groups (alpha-FP greater than 400 ng/ml, alpha-FP less than 400 ng/ml) (r = 0.078, 0.064). The sensitivity for HCC using alpha-FP level alone (greater than 400 ng/ml) was only 52%, and the sensitivity using alpha-AT level alone (greater than 3.2 mg/ml) was 76% of the 46 patients. Combining both tests, sensitivity was improved only to 80%.


Epitope-specific induction of mesangial lesions with proteinuria by a MoAb against mesangial cell surface antigen. A MoAb 1-22-3 (IgG3) was produced in mice immunized with rat glomeruli. A blotting study indicated the antigen molecule recognized by this MoAb has a molecular weight of about 25 kD, which is the same as that of the Thy 1.1 molecule. This MoAb is capable of inducing the morphological changes similar to those induced by anti-thymocyte serum or the anti-Thy 1.1 MoAb, ER4 and massive proteinuria in rats by a single i.v. injection. Proteinuria started immediately after MoAb 1-22-3 injection and peaked on day 5. Reaction products were detected by immunoelectron microscopy in vitro on the limited mesangial cell surface facing endothelial cells and in vivo in partially lysed mesangial cells 30 min after injection. Unlike the proteinuria-inducing MoAb ER4, reactivity of MoAb 1-22-3 was detected neither on endothelial cells, epithelial cells, nor along the glomerular basement membrane in vivo and in vitro. There is a difference in reactivity toward guinea-pig and rabbit materials between MoAb 1-22-3 and the commercial anti-Thy 1.1 MoAb (OX7). The antigenic determinant of MoAb 1-22-3 is concluded to be a new epitope and only the binding of MoAb 1-22-3 to this epitope proved to lead to an abnormal increase of glomerular capillary wall permeability.


The expression and role in T cell adhesion of LFA-3 and ICAM-2 on human thyroid cells. Thyroid follicular cells from patients with Graves' disease and Hashimoto's thyroiditis express intercellular adhesion molecule-1 (ICAM-1) and this is in part responsible for T cell adherence in vitro. To assess the potential role of other adhesion molecules in autoimmune thyroiditis, we investigated the expression and function of lymphocyte function-associated antigen-3 (LFA-3) and ICAM-2 on thyroid cells. Under basal culture conditions, a mean of 22.7% of Graves' thyroid cells (n = 8) expressed LFA-3 and this was enhanced by a mixture of T cell-derived cytokines and by IL-1, but not by TSH. LFA-3 was also demonstrated on Graves' (n = 4) and Hashimoto (n = 2) thyroid cells by immunohistochemical staining ex vivo. A small number of thyroid cells (mean 5.5%, n = 5) expressed ICAM-2 by flow cytometry but this was not altered by cytokines, and ICAM-2 could only be demonstrated on endothelial cells by immunohistochemical staining. It seems likely that contamination of primary thyroid cultures by such cells accounted for the small number of ICAM-2+ cells found using flow cytometry. Almost all of the cultured cells expressing LFA-3 or ICAM-2 also expressed ICAM-1, as assessed by dual staining. Blocking LFA-1, LFA-3, and ICAM-1 with monoclonal antibodies inhibited the adherence of T cells to thyroid follicular cells in assays of cell clustering; antibodies against ICAM-2 had no effect. These results show that two important adhesion receptor ligands, ICAM-1 and LFA-3, are expressed by thyroid cells in autoimmune thyroiditis and that these are likely to have functional importance in allowing T cells to bind to thyroid cell targets. This may play an important role in the initiation and maintenance of Graves' disease and Hashimoto's thyroiditis.


VLA family in rheumatoid arthritis: evidence for in vivo regulated adhesion of synovial fluid T cells to fibronectin through VLA-5 integrin. Adhesion of T cells to extracellular matrix (ECM) proteins through VLA integrin receptors is crucial for lymphocyte trafficking, tissue localization and inflammatory function. We have investigated the expression of different VLA integrins (VLA-1-5) on peripheral blood (PB) and synovial fluid (SF) T lymphocytes from patients with rheumatoid arthritis (RA). Their expression on different cell types from synovial membrane (SM) is also reported. The role of VLA-4 fibronectin (FN) receptors in the interaction of activated SF T cells from RA patients with a 38-kD fragment of FN has been previously demonstrated. Here we have focused functional studies on VLA-5 as an alternative FN receptor for RA T cells. A significant higher proportion of SF T cells were able to bind to an 80-kD fragment of FN, containing the Arg-Gly-Asp (RGD) cell binding site, compared with PB T cells. This attachment was almost completely inhibited by anti-VLA-5 MoAbs as well as by RGD peptides. This enhanced capability by SF T cells appears to be independent of the level of the surface expression of the receptor and correlates better with their activation state as determined by the expression of the activation molecule AIM (CD69). The evidence for the expression of VLA heterodimers on both SF and SM cells from RA patients suggests the possible implication of ECM proteins in mediating and perpetuating inflammation in vivo.