[Calcium antagonists--determination of current status and therapeutic significance]. The important group of drugs known as calcium antagonists is surveyed from a clinico-pharmacological viewpoint. The recently introduced substances are presented in a detailed manner. Common features and differences in pharmacodynamics and pharmacokinetics of the individual calcium antagonists are explained and their differential therapeutic use in the treatment of chronic cardiovascular diseases is discussed. The article also reports on drug interferences, contraindications and possible extensions of indications. An assessment is made of results achieved so far when applying calcium antagonists in the treatment of chronic cardiovascular diseases and in the prevention of arteriosclerosis.


[Results of prostatectomy in the treatment of obstructive benign prostatic hypertrophy]. Aimed to evaluate the results of surgery in patients with obstructive prostatic hypertrophy, we studied the clinical and flow measure results from 51 patients undergoing prostatectomy (31 TUR, and 20 retropubic prostatectomies). It was shown that prostatectomy significantly improved the irritation (63% of patients), and most importantly, the obstructive (89%) symptoms. Peak flow increased in 100% of patients while maximum flow percentile in 92% of patients. Post-operative results showed no correlation to the patients' age, duration of evolution or severity of urinary symptoms, weight of prostate removed or histology of the prostate. Our results were compared with those obtained with other therapeutical options, and it was demonstrated that prostatectomy (both retropubic and RUT) are, clinically and urodynamically, the most effective procedure in the treatment of obstructive prostate hypertrophy.


Codfish allergy in adults. Identification and diagnosis. Ten adult patients with a medical history of immediate allergic reactions following ingestion of minute amounts of codfish were examined together with 27 control subjects (8 nonatopics, 7 pollen allergics, and 12 suffering from atopic dermatitis) all regularly ingesting codfish without experiencing reactions. All 37 subjects were evaluated by skin prick test, RAST, and histamine release test in order to determine the value of these commonly used diagnostic tests. The results were compared to titrated, oral, double-blind, placebo-controlled food challenges (DBPCFCs) with fresh codfish undertaken in the 10 patients, whereas all control subjects were classified as challenge-negative according to their questionnaires. Totally, 7 of 10 medical histories were confirmed by DBPCFC, and in these most of the reported symptoms proved reproducible. Oropharyngeal itching and swelling occurred as the first symptom, always preceding the onset of any accompanying symptom. For identification of DBPCFC-positive subjects, skin prick test and RAST proved to be the most sensitive tests (7/7), and all tests showed specificities of 90-97%. The skin prick test still seems reliable but does not, however, exclude the need for DBPCFC.


[First results of our experience with the Fabian's prosthesis]. Report on our experience with Fabian's prothesis as alternative therapy to benign prostate hyperplasia (B.P.H.) surgery. Throughout 1990 ten patients with various surgical contraindications were implanted the above device. In spite of the series small size (10 patients) and short follow-up (4 to 11 months), we believe the 70% good results obtained is very encouraging and can either be of help to banish the ominous indwelling vesical catheter, a permanent evidence of Urologists' failure in the prostate obstructive disease, or it will allow us to relief patients with B.P.H. non-secondary to other more aggressive therapies.


Basophil histamine release in patients with birch pollen hypersensitivity with and without allergic symptoms to fruits. Histamine release (HR) studies were performed in 40 birch pollen-allergic patients (positive case history, positive SPT, positive birch pollen-specific serum IgE: RAST > or = 3) with (n = 20, A) and without (n = 20, B) fruit hypersensitivity, and 10 nonatopic volunteers (C). Several fruit allergens were used and characterized by protein determination and immunoblot techniques. Dose-dependent HR (apple peel = apple pulp > peach = cherry) was demonstrated in both allergic groups, but to a higher extent in patients with fruit allergy (P < 0.01). Increased basophil sensitivity to birch pollen was found in the group with fruit allergy (P < 0.001). Strong correlations between the mediator response induced by several fruits indicate common allergens within the extracts. We conclude that fruit-related symptoms require not only high specific serum IgE, but a strong cellular sensitization to birch pollen allergens together with an increased cellular reactivity to fruit allergens.


Effect of astemizole on antigen-mediated histamine release from the blood of patients with allergic rhinitis. The main objective of this study was to test the effectiveness of astemizole in vitro in blocking the release of histamine from blood of patients with allergic rhinitis. The results of this investigation indicated that astemizole inhibited allergen-mediated histamine release from blood basophils of patients with this allergic disorder. The inhibition by astemizole (33-156 mumol) was immediate, requiring no pre-incubation of the cells, and was dose-dependent, with maximal inhibition of about 91%. The relatively high potency of astemizole in inhibiting the immunologic release of histamine may provide an additional measure in the treatment of allergic rhinitis with this H1-receptor antagonist.


Histamine release in immediate-type hypersensitivity reactions in intact human skin measured by microdialysis. A preliminary study. The purpose of this study was to evaluate the application of a microdialysis technique for measurement of interstitial histamine levels in intact human skin. Three allergic subjects were investigated. Single dialysis fibers were glued to nylon tubings and inserted in forearm skin by means of a fine cannula. Dialysis fibers were inserted in triplicate and perfused with isotonic saline at a rate of 3 microliters/min. After a period of 2 h a 60-microliters base-line period was established. Then the patients were skin prick tested (SPT) with allergen in duplicate and a single saline control. Dialysate was collected in consecutive 30 microliters fractions. Histamine concentration in the dialysate was analyzed with a glass fiber fluorescence assay. Median base-line histamine level was 4 (range 4-7) ng/ml. Following allergen SPT, dialysate histamine concentration increased to 81 ng/ml (74-128), with maximum values 10-20 min after SPT. Intraindividual coefficient of variation on peak histamine levels was 18.9%. No histamine increase was seen following saline SPT. We consider microdialysis to be a valuable method for assessment of allergic mechanisms in intact human skin.


Contact allergy to budesonide contained in a nasal spray. We describe a case of contact allergic sensitization to budesonide in a nasal spray used in the treatment of allergic rhinitis. No cross-sensitivity to other topical corticosteroids was found.


[Comparison of the effects of epsilon-aminocaproic acid and aprotinin on intra- and postoperative bleeding in heart surgery]. Excessive bleeding during and after cardiac surgery with cardiopulmonary bypass is a real problem in this kind of surgery. The use of prophylactic high doses of aprotinin (APROT) reduces blood loss in this context but this treatment is expensive. Some investigators have advocated that epsilon-aminocaproic acid (EACA), a cheaper antifibrinolytic drug, could reduce blood loss in cardiac surgery. The goal of this prospective study was to determine if EACA is as effective as APROT for this clinical condition. Sixty patients undergoing elective surgery for cardiac disease were randomly allocated to one of the two groups. Drugs were administered after induction of anesthesia at a dose of 2.10(6) UIK in the APROT group or 5 g in the EACA group. The same dose was added to the priming of the cardiopulmonary bypass circuit. Until the skin closure the patients received 5.10(5) UIK/h of APROT or 2 g/h of EACA. Bleeding during and after surgery was not different between the two groups. No complication, directly due to the treatment administered, was observed. EACA seems to be as effective as APROT to reduce intra and post cardiac surgery blood loss. EACA has the advantage of being cheaper (treatment is approximately 200 times cheaper), therefore allowing a wider use.


The changing pattern of infantile intussusception in northern Nigeria: a report of 47 cases. Infantile intussusception is not rare in Africa, but was reported to be relatively uncommon. This retrospective review of 47 infants with the diagnosis of intussusception included 38 boys and nine girls. Surprisingly, the clinical presentation is at variance with previous reports and beliefs about intussusception in tropical Africa, but is similar to those from Europe and North America. Contrary to expectation, about 70% of infants presented with the classical clinical picture of intussusception with the complete features of vomiting, 'colicky abdominal pain', blood mucoid stools and palpable abdominal mass. The other 30% presented with various combinations of the components of the tetrad. The small bowel was the leading point in 30 infants and the colon in 17. Of the small bowel variety, the two main types were the ileo-ileal and the ileocolic, while the ileocaecal and the various types of colo-colic intussusception were of large bowel origin. Morbidity was minimal. There were four deaths. Intussusception in this series differs in many respects from previous experience in Zaria.


Evaluation of malto-dextrin/glycine oral rehydration solution. The efficacy and safety of malto-dextrin/glycine-based oral rehydration solution (ORS) when compared with the glucose-based oral rehydration solution (WHO) was evaluated in a randomized double-blind clinical trial. Thirty-one subjects and 31 controls were studied. The mean values of the ORS intake, stool output, duration of diarrhoea, urine output, weight gain and serum electrolytes were comparable in both the study and the control groups (p > 0.05). It was found that the malto-dextrin/glycine ORS offered no therapeutic advantage over the standard glucose ORS (WHO). In view of the accidental preponderance of children of significantly lower nutritional status among the control group (p < 0.05), it is likely that the efficacy of malto-dextrin/glycine ORS might actually be less than that of the glucose ORS.


Is cow's milk suitable for the dietary supplementation of rural Gambian children? 1. Prevalence of lactose maldigestion. To define the age-related prevalence of lactose maldigestion (LM), 218 children aged 13-72 months from a rural Gambian village were studied using a lactose breath hydrogen test. There was a significant rise in LM from 21% to 76% between the 2nd and 3rd-5th years of life (p < 0.001). Only eight children had diarrhoea within 2 weeks before the test. Diarrhoea and abdominal discomfort followed in only seven children (3%). There was no difference in mean time of introduction of supplementary diet or cessation of breastfeeding between children aged 12-36 months with lactose digestion and LM. There was a greater proportion of children with normal lactose digestion than with LM in those still receiving breast milk (85% vs 15%), and a greater proportion of LM in the fully weaned group (63% vs 37%) (p < 0.001). There was no relation between weight-for-age or weight-for-height and the ability to digest lactose. The ability to digest lactose is lost in the majority of children during the 2nd year, coincident with the cessation of breastfeeding, but is not a major factor associated with growth failure thereafter.


Is cow's milk suitable for the dietary supplementation of rural Gambian children? 2. Patterns of cow's milk intake. Cow's milk has been advocated as a source of supplementary diet for children in many parts of the developing world. The frequency and forms of cow's milk intake and the factors regulating its availability and consumption by 507 children aged up to 6 years in three Gambian villages were measured. Nineteen per cent of mothers did not give cow's milk to their children. Of 413 children taking cow's milk, 41% consumed it only once a week, 32% two to four times a week, 19% once a day and 8% more than once a day. Eighty per cent of children took both fresh and sour milk. Forty-one per cent of infants up to 1 year old received cow's milk at some time. There was a decrease with age in the proportion of children taking fresh milk and a rise with age in the proportion taking sour milk alone (p < 0.001) and both forms of milk. Only 2% of children were reported to have an adverse reaction to fresh cow's milk. The main factors affecting intake were the availability of money and milk. The consumption of cow's milk in early life is common and free of adverse effects. When it is available, it should be used to supplement the diet of the weaned child.


Purulent meningitis in children: outcome using a standard management regimen with chloramphenicol. We report a prospective study of 108 children aged from 2 weeks to 10 years with purulent meningitis admitted to the children's wards of Port Moresby General Hospital, 105 of whom were treated with a standard management regimen using chloramphenicol (given intramuscularly initially) as the only antibiotic. The case fatality rate of 16.7% and the apparently low morbidity rate were felt to be very satisfactory in a high risk population. The medical officers' compliance with the standard antibiotic regimen was good, but was disappointing for the use of anticonvulsants. The presence of convulsions before admission or at any time in the illness, coma on admission, and a history of illness of more than 3 days duration prior to admission were strongly associated with death. We believe phenobarbitone should be given prophylactically to children less than 2 years of age who have meningitis. Further improvements in outcome are likely to be achieved not by changes in antibiotic policy but by improving early diagnosis and basic supportive care, and by preventing convulsions.


Indications for lumbar puncture in children presenting with convulsions and fever of acute onset: experience in the Children's Emergency Room of the University of Benin Teaching Hospital, Nigeria. A total of 522 children, aged 1 month to 6 years, who presented with convulsions and fever of acute onset at the Children's Emergency Room of the University of Benin Teaching Hospital over a 1-year period, were prospectively evaluated. Bacterial meningitis was diagnosed in 22 (4.2%) on bacteriological and/or biochemical evidence. The causative organisms were cultured from the CSF in 13 (Neisseria meningitidis = 7, Streptococcus pneumoniae = 5 and Haemophilus influenzae = 1) and identified by Gram stain only in three (Gram-positive diplococci = 2 and Gram-negative diplococci = 1). No organisms were identified in the CSF of six of the children with meningitis. The prevalence of meningitis declined sharply after 6 months of age. Six of the children with bacterial meningitis lacked classical meningeal signs but had other indications for lumbar puncture. The following were significantly associated with meningitis: age under 6 months; focal or multiple seizures; absence of a past or family history of seizures; unrousable coma; and an extracranial focus of infection. It is concluded that bacterial meningitis occurs in a good proportion of children, even beyond infancy, with convulsions associated with fever of acute onset, and that decision on the need for lumbar puncture should be guided by clinical features such as age and the presence of complex febrile seizures.


Screening for glucose-6-phosphate dehydrogenase deficiency can prevent severe neonatal jaundice. Infants with the severe variant of glucose-6-phosphate dehydrogenase (G6PD) deficiency may develop hyperbilirubinaemia sufficiently severe to cause kernicterus and death, acute haemolysis on exposure to oxidant stress, congenital non-spherocytic haemolytic anaemia and, rarely, increased susceptibility to bacterial infection. In spite of these potential problems, G6PD deficiency is often not included among screening programmes for inherited disorders. In a comprehensive screening and educational programme, we tested around 34,000 infants for G6PD deficiency. Of the total group, 18.4% (24.5% boys and 11.8% girls) were deficient. Forty-two of the 6246 (0.67%) G6PD-deficient infants required exchange transfusion. None of them developed kernicterus. By contrast, of 4755 infants who had not been screened because they were born at home, three developed kernicterus. In addition, four G6PD-deficient infants had developed kernicterus in the 20-month period prior to the screening programme. None of the hyperbilirubinaemic infants had blood group incompatibility or any other identifiable cause of hyperbilirubinaemia. To avoid this disastrous result, we believe that neonatal screening for G6PD deficiency, together with a comprehensive education programme, is advisable in those parts of the world where the severe variant of G6PD deficiency is prevalent.


Acquired immunodeficiency syndrome: education exposure, knowledge and attitude of Nigerian adolescents in Calabar. The knowledge of and attitude to acquired immunodeficiency syndrome (AIDS) of 738 secondary school youths in Calabar were studied by questionnaire in March 1991. Most (92%) of the adolescents had heard about AIDS, largely through the mass media (79-85%), with parents and teachers contributing in less than 40%. About 30% did not know that AIDS exists in Nigeria. Most of them knew that promiscuity, blood transfusion and sharing injection needles and syringes are the major modes of transmission, but a number still incriminate toilet seats, eating utensils, hand-shaking and kissing. Only 31% were aware that condoms provide protection. For self-protection the youths prefer abstinence (45%) and confinement to one sex partner (19%). Only 3.6% would adopt the use of condoms. To prevent the spread of AIDS, the youths prescribe isolation (37%), treatment (34%), and killing (14%) of diagnosed cases. Most youths (77%) will stop seeing friends and 63% will reject relatives who develop AIDS. The study reveals that while general awareness of AIDS is fairly good, detailed knowledge is riddled with misconceptions and confusion. It is recommended that physicians in the community should assist in disseminating accurate information, with the support of parents, teachers and the youths themselves.


Preterm infants with patent ductus arteriosus: treatment with an enteral preparation of indomethacin. Forty-one preterm infants with a mean gestational age and birthweight of 29.7 weeks and 1322 g, respectively, were treated with an enteral preparation of indomethacin for the closure of haemodynamically significant patent ductus arteriosus. The indomethacin was prepared by diluting the content of the commercially available 25-mg indomethacin capsule with lactose by the method of trituration. The mixture was suspended in solution form just prior to administration to the infants. The ductus arteriosus was successfully closed in 29 infants, giving an overall success rate of 71%. The response to treatment was considered satisfactory, except for infants whose birthweights were less than 750 g. Only two infants (5%) developed major complications, both in the form of ileal perforation. As the outcome of treatment with this enteral preparation of indomethacin was comparable with that of the intravenous preparation reported in previous studies, it would be an acceptable alternative to the intravenous preparation in areas where the latter is not available or considered expensive.


Rett syndrome in South Africa.

Rett syndrome in South Africa. Rett syndrome is a fairly recently recognized neurodevelopmental disorder of unknown aetiology that affects exclusively girls in whom early development is apparently normal but by the age of 6-18 months autistic behaviour and dementia, apraxia of gait, stereotypic repetitive hand movements, seizures and deceleration of head growth occur. Except for one Brazilian case, all previous reported cases have been from the northern hemisphere. We report three children (two Indian and one African) with the clinical features of Rett syndrome and believe these to be the first documented cases in Indian and African children in the southern hemisphere. More widespread knowledge of this syndrome is required, especially in developing countries, in order to prevent unnecessary and costly investigations, and to help families deal effectively with this important syndrome.


Giant lipoblastoma: a case report.

Giant lipoblastoma: a case report. A Nigerian boy presented to the paediatric surgical unit of this institution with what appeared to be a malignant congenital mesenchymal neoplasm of the right axilla. The mass was lipofibromatous and invaded local structures. Excision was complete. The specimen weighted 4 kg. Histological features were those of a lipoblastoma. Following excision, he had a neurological deficit involving the 4th and 5th cervical neural segments which persisted till he was lost to follow-up 2 years later.


The association of household pollutants and socio-economic risk factors with the short-term outcome of acute lower respiratory infections in hospitalized pre-school Nigerian children. In a 9-month study of acute lower respiratory infections (ALRI), the short-term prognostic implications of socio-economic and household risk factors were examined in 103 hospitalized pre-school Nigerian children. Seventy-nine (77%) subjects were potentially exposed to the combustion products of kerosene stoves, 16 (16%) to wood smoke and five (5%) to the products of cooking gas combustion. Only 17 subjects (17%) were exposed to household cigarette smoke. A highly significant association (p < 0.005) was shown between household cooking fuel and the outcome of hospitalization: five (63%) of the eight who died were potentially exposed to wood smoke. The duration of hospitalization was only significantly associated with paternal income (p < 0.05). None of the other domestic risk factors was significantly related to outcome or duration of admission. These findings suggest an association between an adverse outcome of ALRI and domestic exposure to wood smoke. While the hospital-based source will not allow definite conclusions, the present findings underscore the need for community-based studies. The implications for future strategies of ARI control are discussed, with suggestions.


Bacteraemia in severely malnourished children. During a 2-year period, 792 severely malnourished children (kwashiorkor, marasmus and marasmic kwashiorkor) were hospitalized (14.8% of all admissions). Community-acquired bacteraemia was recorded in 7.7% and nosocomial bacteraemia in 2.2% of malnourished children. The relative risk for bacteraemia on admission was 1.6 (95% confidence interval [CI] 1.3-1.9) and 2.0 for nosocomial bacteraemia (95% CI 1.4-3.0) in malnourished children as compared with well nourished children. Bacteraemia was 3.5 times more common in children with kwashiorkor than in marasmic children. Gram-negative enteric aerobes were isolated 2.6 times more frequently from malnourished than from well nourished children. The relative risk of death in malnourished children with bacteraemia was 2.5 times that of malnourished children without bacteraemia. Thirty-six per cent of deaths in children with kwashiorkor were related to bacteraemia. This study emphasizes the increased risk of bacteraemia in malnourished children, particularly those with kwashiorkor, and the impact on mortality.


Plasma levels of mebendazole in children with hydatid disease. The plasma levels of mebendazole in children with hydatid cyst disease were measured with high pressure liquid chromatography. In 24 children who received mebendazole orally in a dose of 50 mg/kg, the mean (SD) level 4 hours later was 25.76(9.81) ng/ml (87.6(33) nanomole/l). This result was similar to those in most adult series. During more prolonged treatment, the plasma level 4 hours after the dose of the drug rose significantly with respect to the initial level (p < 0.05).


Neonatal and post-neonatal onset of early congenital syphilis: a report from Mozambique. Congenital syphilis (CS) has been and continues to be a principal public health problem in developing countries. Despite the wide experience acquired, physicians still have problems in diagnostic evaluation. We report 145 cases of CS at the Central Hospital, Maputo, emphasizing the differences in clinical features and in the results of serological and X-ray examinations between the neonatal and post-neonatal age groups. In the post-neonatal age group, the clinical expression of CS is mostly overt. It is commonly recognized that manifestations of CS in the neonatal age group are often poor or negative, yet a relevant percentage of CS that we report were fully symptomatic. In the neonatal age, the Venereal Disease Research Laboratory (VDRL) test in the mother and characteristic osteochondritic lesions on X-ray examination of the long bones help to make the diagnosis; in the post-neonatal age group, the VDRL test in the child is more often positive than in the mother and X-ray examination shows most periostitic lesions.


Pyogenic liver abscess causing acute Budd-Chiari syndrome. Acute Budd-Chiari syndrome caused by compression of the inferior vena cava by a space-occupying lesion in the liver is rare in children. We report a case where the compression was due to a large staphylococcal abscess in the right lobe of the liver. A high index of suspicion in such a case ensures early intervention and saves life.


Hyponatraemia and the inappropriate ADH syndrome in pneumonia. We studied serum sodium, plasma osmolality and urinary sodium and osmolality on days 1, 3 and 5 of hospitalization of 100 children aged from 1 month to 12 years admitted with a diagnosis of pneumonia. Hyponatraemia (serum sodium concentration < or = 130 mmol/l) was found in 31 patients at the time of admission. The probable cause of hyponatraemia in 94% of cases was the syndrome of inappropriate antidiuretic hormone secretion (SIADH). Symptoms and signs indicative of severe pneumonia were two to three times more frequent and the mean duration of tachypnoea, chest-wall retraction and hospital stay about one and a half times longer in children with hyponatraemia. Four children died (two on day 1, one on day 5 and one on day 8); all four had a serum sodium concentration < or = 125 mmol/l which persisted until death. Of the remaining 27 hyponatraemic children, serum sodium concentrations returned to normal on day 3 in 26, while in one hyponatraemia persisted until day 7. The recovery from hyponatraemia showed a good correlation with improvement in clinical signs of respiratory distress. The SIADH occurred in about one-third of the children hospitalized for pneumonia, and was associated with a more severe disease and a poorer outcome. Perhaps fluid restriction in these cases may improve the outcome.


Increase in total serum triglyceride and phospholipid in kwashiorkor. Fasting serum lipids in children presenting with protein-energy malnutrition were studied in comparison with those of sex- and age-matched well nourished controls. There was no difference in serum total cholesterol between malnourished and well nourished children. However, serum triglyceride and phospholipid concentrations were significantly higher in children with kwashiorkor or marasmic kwashiorkor than in well nourished controls. It appears that serum concentrations of triglycerides and phospholipids increase with increasing severity of malnutrition, and that fasting serum triglyceride and phospholipid concentrations above 3 mmol/l are jointly predictive of a poor prognosis in malnourished children.


Nutritional pattern and eco-physiology of Hortaea werneckii, agent of human tinea nigra. The life cycle of Hortaea werneckii includes yeast-like, hyphal and meristematic growth. The preponderance of each form of propagation can be influenced by environmental conditions. The clinical entity 'tinea nigra' is explained by ecological similarities between supposed natural niches and human hyperhydrotic skin. The species is recognizable by assimilation of lactose, nitrate and nitrite, no or little growth with L-lysine, cadaverine, creatine and creatinine, and tolerance of 10% NaCl. It generally does not grow at 36 degrees C.


Role of prostanoids in the inflammatory reaction and their therapeutic potential in the skin. Vasodilatory prostaglandins (PG), contributing to the inflammatory reaction, have gained considerable attention. It is becoming apparent that PG have pharmacological effects traceable to biological activities distinct from smooth muscle relaxation. The data from pharmacological experiments presented here indicate the diverse action of vasodilatory PG analogues in the skin of laboratory animals. Nocloprost, a stable PGE2 analogue, induced erythema in intact skin of rats when applied topically and inhibited in the same dose range an irritant-induced inflammatory reaction in the ears of mice. Iloprost, a stable PGI2 analogue, showed proinflammatory activity after local application by enhancing the leukotriene B4 induced cell infiltration in the skin of mice. The attenuation of the spreading of ear necrosis in mice, on the other hand, indicates an anti-ischemic therapeutic potential of iloprost. Research in the past has elucidated the influence of PG on the vascular component of inflammation, but the role of PG on the cellular component of inflammation is less clear. The diverse effects of PG in skin indicate the need for a better understanding of their local actions.


Effects of muscarinic receptor agonists and antagonists on rat brain serotonergic activity. The effects of some muscarinic M1 and M2 receptor agonists and antagonists on rat brain serotonergic activity was assessed by noting their effects on the levels of 5-hydroxytryptamine (5-HT) and its major metabolite, 5-hydroxyindole acetic acid (5-HIAA), estimated by a high pressure-liquid chromatographic (HPLC) technique. The muscarinic M1 receptor agonists, arecholine and McN-A-343, and the M2 receptor agonists, gallamine and AF-DX 116, induced a dose-related decrease in the concentrations of both 5-HT and 5-HIAA. On the contrary, scopolamine and the selective M1 receptor antagonist, pirenzepine, increased the levels of the amine and its metabolite. The anti-cholinesterase agent, physostigmine, and the putative M2 receptor agonist, carbachol, induced a dose-related dual effect, with the smaller doses decreasing and the higher doses increasing 5-HT and 5-HIAA concentrations. The results indicate that an inverse relationship exists between the cholinergic and serotonergic neurotransmitter systems in the rat brain due to the likely presence of muscarinic heteroreceptors on serotonergic neurones. The data also indicates that though physostigmine and carbachol may function as M2 receptor agonists, they lose their receptor specificity on dose increment.


Repeated batch cultivation of rBHK cells on Cytodex 3 microcarriers: antithrombin III, amino acid, and fatty acid metabolic quotients. Anchorage-dependent human antithrombin III-producing recombinant baby hamster kidney (rBHK) cells were cultivated on Cytodex 3 microcarriers in repeated batch mode. During a 3-month experiment four different low-serum (0.025% fetal bovine serum) or serum-free medium formulations were evaluated for (a) the initial growth phase of cells and (b) the subsequent production phase, whereby two free fatty acid (FFA) supplements were examined with respect to their growth-promoting and product-formation-enhancing properties. Selected nutrient and (by)product consumption and production rates (including those for antithrombin III, amino acids, and fatty acids) are reported. The calculated metabolic quotients reflect the prevailing slow growth conditions (mu approx. 0.06 day-1) associated with microcarrier cultures. Specific antithrombin III productivities vary significantly as a function of the feed medium supplementation with FFA.


The Hopkins Lupus Pregnancy Center: 1987-1991 update. The course of pregnancy in patients with systemic lupus erythematosus is not known. The Hopkins Lupus Pregnancy Center has followed 64 patients (74 pregnancies) prospectively since 1987. Patients are seen monthly and clinical and pregnancy-related data collected, with particular emphasis on the occurrence of lupus flare. Flare rate during pregnancy was 1.63 per person-year, compared to 0.64-0.65 after delivery or in non-pregnant patients. Flare did not influence pregnancy outcome. Low serum C3 or C4 and high anticardiolipin antibody predicted pregnancy loss, and prednisone dose, aspirin use, diastolic second trimester blood pressure, C3 at first visit, and race predicted preterm birth. Maternal flare and preterm birth are important risks in lupus pregnancy. The latter can be predicted from maternal pregnancy data.


[A case of primary intracranial malignant melanoma showing leptomeningeal dissemination]. A 28-year-old woman was hospitalized in drowsy state with signs of increased intracranial pressure. CT scans revealed diffuse increased density with marked enhancement in the subarachnoid space, as well as ventricular dilatation. V-P shunt operation was performed to control intracranial pressure. Repeated cytological examinations of CSF couldn't determine the tumor origin. CT scan of thoracic spine showed a cystic tumor in its dorsal aspect. T2-weighted MRI revealed multiple spotty low intensity, specific to melanin granules, throughout the whole spine. Her thoracic spine was explored, and the intradural tumor was partially removed. Histopathological examination revealed the tumor cell which had dark nucleus with conspicuous nucleolus and cytoplasmic granules. These findings were compatible with malignant melanoma. Her general condition were deteriorated progressively and she died about 5 months after her admission. Postmortum examination showed diffuse leptomeningeal invasion of dark tumor throughout the entire central nervous system, and metastasis to peritoneum and omentum via V-P shunt system. Histopathological examination proved the tumor to be malignant melanoma. Electrone microscopic examination also revealed melanosome in the cytoplasm. Primary intracranial malignant melanoma is divided in two groups, nodular type and leptomeningeal type. In the latter type, early diagnosis is very difficult, just as in our case, because only a little tissue specimen can be obtained. In a case of leptomenigeal carcinomatosis, possibility of primary malignant melanoma, though rare, should always be kept in mind, and specific staining such as Fontana-Masson's staining should be tried.


Evaluation of a microcarrier process for large-scale cultivation of attenuated hepatitis A. Microcarrier culture was investigated for the propagation of attenuated hepatitis A vaccine in the anchorage-dependent human fibroblast cell line, MRC-5. Cells were cultivated at 37 degrees C for one to two weeks, while virus accumulation was performed at 32 degrees C over 21 to 28 days. The major development focus for the microcarrier process was the difference between the cell and virus growth phases. Virus antigen yields, growth kinetics, and cell layer/bead morphology were each examined and compared for both the microcarrier and stationary T-flask cultures. Overall, cell densities of 4-5 x 10(6) cells/ml at 5-10 milligrams beads were readily attained and could be maintained in the absence of infection at either 37 degrees C or 32 degrees C. Upon virus inoculation, however, substantial cell density decreases were observed as well as 2.5 to 10-fold lower per cell and per unit surface area antigen yields as compared to stationary cultures. The advantages as well as the problems presented by the microcarrier approach will be discussed.


Competition of Mn2+ and Zn2+ with 59Fe2+ and 59Fe3+ for the plasma membrane receptors from lactating mouse mammary gland. Nonlabeled MnCl2 and ZnSO4 compete with 59Fe(2+)-ascorbate and 59Fe2(3+)O3 for transport binding sites situated on the plasma membranes of lactating mouse mammary gland cells. The binding was found to be a process reaching saturation. The heterologous competition used here ruled out the participation of transferrin and to propose that Fe, Mn, and Zn are transported from blood to milk by a mechanism involving one receptor during lactation. Further experiments are necessary to establish the details of the transport mechanism.


Tissue antioxidant status in streptozotocin-induced diabetes in rats. Effects of dietary manganese deficiency. Interactions between manganese (Mn) deficiency and streptozotocin (STZ)-diabetes with respect to tissue antioxidant status were investigated in male, Sprague-Dawley rats. All rats were fed either a Mn-deficient (1 ppm) or a Mn-sufficient (45 ppm) diet for 8 wk. Diabetes was then induced by tail-vein injection of STZ (60 mg/kg body weight), after which the rats were kept for an additional 4 or 8 wk. The control groups comprised rats not injected with STZ and fed either Mn-deficient or Mn-sufficient diets for a total of 12 wk. The Mn-deficient diet decreased the activities of manganese superoxide dismutase (MnSOD) in kidney and heart, and of copper-zinc superoxide dismutase (CuZnSOD) in kidney, in the non-diabetic animals. In the diabetic rats, the Mn-deficient diet induced more pronounced decreases in activities of these same enzymes, and also increased liver MnSOD activity. Plasma and hepatic vitamin E levels increased progressively with the duration of diabetes, independent of dietary Mn intake. Lipid peroxidation, as measured by H2O2-induced production of thiobarbituric acid reactive substances in erythrocytes, also increased, concomitant with decreased liver and kidney glutathione (GSH) levels. These findings demonstrate for the first time and interactive effective between Mn deficiency and STZ-diabetes, resulting in amplification of tissue antioxidant changes seen with either Mn deficiency or STZ-diabetes alone. This effect of Mn deprivation in experimental diabetes suggests a physiological role for Mn as an antioxidant nutrient.


Three-dimensional reconstruction of a complex of human alpha 2-macroglobulin with monomaleimido Nanogold (Au1.4nm) embedded in ice. Cysteine 949 and glutamine 952 are known to be part of the thiol ester site of each of the four subunits of human alpha 2-macroglobulin (alpha 2M). The hydrolysis of this thiol ester bound to methylamine results in the incorporation of the amine and liberation of a free sulfhydryl group that can be specifically labeled. Therefore, a high-resolution marker specific for the sulfhydryl groups, the monomaleimido Nanogold (Au1.4nm) cluster was used to bind this amino acid. After cryoelectron microscopy, a three-dimensional reconstruction of the alpha 2M-Nanogold conjugates (alpha 2M-Au1.4nm) was achieved, revealing the internal location of the thiol ester sites in the transformed alpha 2M molecules. From this study we propose three possible locations for the cysteine 949.


Studies of the interaction between boron and calcium, and its modification by magnesium and potassium, in rats. Effects on growth, blood variables, and bone mineral composition. Two experiments were performed to confirm that boron interacts with calcium, and that this interaction can be modified by dietary magnesium and potassium in the rat. Upon manipulating the dietary variables listed above, it was found that under certain conditions, boron and calcium deprivation similarly affected several variables; for example, they both could be made to elevate plasma alkaline phosphatase activity and to depress femur calcium concentration. Under some dietary conditions, both boron and calcium deprivation affected some variables related to blood or iron metabolism. However, the effects of dietary boron and calcium on spleen weight/body weight ratio, hematocrit, and femur iron concentration generally were not similar. Femur copper, magnesium, phosphorus, and zinc also were affected by an interaction between boron and calcium under some dietary conditions. The findings show that there is a relationship between boron and calcium, but they do not clearly indicate the nature of the relationship. However, the data suggest that boron and calcium act on similar systems in the rat.


The effect of time of introduction of a high-fructose, low-copper diet on copper deficiency in male rats. The purpose of the present study was to compare the time of introduction of the high-fructose low-copper diet on the expression of copper (Cu) deficiency. Weanling male rats were randomly assigned to either a diet containing 62.7% fructose or starch, and 6.0 (F+Cu) or 0.6 (S-Cu) microgram Cu/g diet, respectively, for either 1, 2, or 3 wk before being transferred to a diet containing fructose and inadequate in copper (F-Cu). At week 10, body weight and relative heart size of rats initially consuming the F + Cu diet was inversely related to the week placed on the F-Cu diet, but not for those initially consuming S-Cu. Hematocrit, hepatic Cu concentration and RBC superoxide dismutase activity were significantly lower in rats initially consuming S-Cu when compared to those fed F + Cu. Mortality was greatest in rats switched to the F-Cu diet at weeks 1 and 2 when compared to those switched at week 3 regardless of the type of diet initially consumed. Plasma cholesterol, triacylglycerols, and blood urea nitrogen concentrations were not significantly altered by the type of diet initially consumed or by the time of introduction of the F-Cu diet. It was concluded that changing rats to a F-Cu diet at 1, 2, or 3 wk after weaning did not significantly improve some of the characteristic signs associated with Cu deficiency, but the later that the F-Cu diet was introduced after weaning the greater the chances for survival.


Zirconium. An abnormal trace element in biology. The action of Zirconium (Zr) on biological systems presents an enigma. It is ubiquitous, being present in nature in amounts higher than most trace elements. It is taken up by plants from soil and water and accumulated in certain tissues. The entry into animal systems in vivo is related to the mode of exposure and the concentration in the surrounding environment. Retention is initially in soft tissues and then slowly in the bone. The metal is able to cross the blood brain-barrier and is deposited in the brain and the placental barrier to enter milk. The daily human uptake has been known to be as high as 125 mg. The level of toxicity has been found to be moderately low, both in histological and cytological studies. The toxic effects induced by very high concentrations are nonspecific in nature. Despite the presence and retention in relatively high quantities in biological systems, Zr has not yet been associated with any specific metabolic function. Very little information is available about its interaction with the compounds of the genetical systems, such as nucleic acids. Apparently, the metal is neither an essential nor toxic element in the conventional sense. However, the increasing exposure to this element through its increasing use in new materials and following radioactive fallout, has increased the importance of the study of its effects on living organisms. The tetravalent nature of the ionic state and the high stability of the compounds formed are important factors that need to be considered, as also the accumulation of this element in the brain, reminiscent of the relationship between Al3+ and Alzheimer's disease.


Serum nickel levels of diabetic patients and healthy controls by AAS with a graphite furnace. In this study, serum nickel levels of diabetic patients and healthy controls were determined by AAS with a graphite furnace. The serum nickel concentrations were found to be 1.15 +/- 1.89 micrograms/L in healthy controls and 0.82 +/- 0.74 microgram/L in diabetics. There was, however, no statistically significant difference between the two groups (p = 0.13). The relationship of nickel levels to diabetes type and duration, diabetic complications and treatment, sex, age, and heredity was investigated. However, again no significant differences were found, nor was there any correlation between serum nickel levels and blood sugar, HBa1c, fructosamine, sialic acid levels, and age.


Effects of excess selenomethionine on selenium status indicators in pregnant long-tailed macaques (Macaca fascicularis). Forty pregnant long-tailed macaques were treated daily for 30 d with 0, 25, 150, or 300 micrograms selenium as L-selenomethionine/kg body weight. Erythrocyte and plasma selenium and glutathione peroxidase specific activities, hair and fecal selenium, and urinary selenium excretion were increased by and were linearly related to L-selenomethionine dose. Hair selenium was most sensitive to L-selenomethionine dose, with an 84-fold increase in the 300 micrograms selenium/(kg-d) group relative to controls (r = 0.917). Daily urinary selenium excretion (80-fold, r = 0.958), plasma selenium (22-fold, r = 0.885), erythrocyte selenium (24-fold, r = 0.920), and fecal selenium (18-fold, r = 0.911) also responded strongly to L-selenomethionine. Erythrocyte and plasma glutathione peroxidase specific activities increased 154% and 69% over controls, respectively. Toxicity was associated with erythrocyte selenium > 2.3 micrograms/mL, plasma selenium > 2.8 micrograms/mL, and hair selenium > 27 micrograms/g. Plasma, erythrocyte, and hair selenium concentrations may be useful for monitoring and preventing the toxicity of L-selenomethionine administered to humans in cancer chemoprevention trials.


Selenium in Hungary. The rock-soil-human system. Selenium (Se) status of Hungarian rock-soil systems has been poorly investigated. The goal of this study was to get general information on the main features of Se geochemistry, and to have a brief look into the human Se status in Hungary, developing an appropriate method for a forthcoming systematic regional investigation. A multistage program was applied: 1. stream sediment sampling; 2. investigation of rock-soil profiles typical in Hungary; and 3. extraction of mobile Se fractions from soils. Human serum samples from apparently healthy blood donors residing in both urban and rural areas were collected from three districts in Hungary. It was concluded that Hungarian rock-soil systems, especially the acid igneous rocks and the widely distributed young sediments (loess and sand formations) with most of the agricultural activity, are low in Se, and the mean serum Se level of the blood samples were also low.


Two modes of homologous C3 deposition on Ramos Burkitt's lymphoma cell substrains co-expressing DAF (CD55), CD59, and CR2 (CD21), and on cells lacking them. We established decay-accelerating factor (DAF)/CD59-positive and -negative substrains of a human B cell line, Ramos, R(DAF+/CD59+) and R(DAF-/CD59-) respectively. Unexpectedly, treatment of R(DAF+/CD59+) cells with Mg2(+)-EGTA-serum resulted in efficient C3 deposition, while treatment of R(DAF-/CD59-) cells did not. All six substrains of R(DAF-/CD59-) cells were CR2-negative, and treatment of the cells with M177 [a membrane cofactor protein (MCP) cofactor-blocking antibody] and/or acidic buffer only minimally affected the extent of C3 deposition. However, when R(DAF-/CD59-) cells were pretreated with M177 followed by incubation with low conductivity (3 mS) Mg(2+)-EGTA-serum, C3 deposition leading to effective cytolysis was provoked. On the other hand, all seven R(DAF+/CD59+) substrains were CR2-positive and could potentially induce C3 autoactivation without cytolysis under physiological conditions. Both M177 and pH again minimally affected the extent of C3 deposition. However, conductivity altered the sensitivity to C3: at under 3.0 mS, R(DAF+/CD59+) cells became almost insensitive to alternative pathway-mediated C3 deposition. Anti-CR2 partially inhibited C3 deposition on R(DAF+/CD59+) cells and C3 deposition was abrogated on the CR2-lacking R(DAF-/CD59-) cells, suggesting that CR2 was associated with the deposition of C3. These results, together with the finding that fluid phase activation of complement did not enhance C3 deposition, suggest that there are two distinct modes of spontaneous homologous C3 deposition on human lymphoma cells. In one case, CR2 or its related molecules participates in C3 deposition overcoming the protective function of DAF/MCP and this type of C3 deposition is maximized under physiological conditions. In the other case, C3 deposition is induced by another homologous C3 activator that becomes functional under low conductivity conditions and the absence of DAF/MCP. These two modes of homologous alternative pathway activation would explain the reported instances of spontaneous C3 deposition on human B lymphoid cell lines (under physiological conditions in the presence of DAF/MCP) and on paroxysmal nocturnal hemoglobinuria erythrocytes (under low conductivity conditions in the absence of DAF/MCP).


Agonist-antagonist interactions in the skin: comparison of effects of loratadine and cetirizine on skin vascular responses to prick tests with histamine and substance P. The skin vascular responses (weal, flare, blood flow measurements) elicited by intradermal administration by pricking of histamine (HS) and substance P (SP) were evaluated 6 h after a single intake of anti-H1 agents displaying different activity profile on skin tests at currently recommended dosages (loratadine 10 mg, cetirizine 10 mg) as compared to placebo (P). The weal and flare response and the increases of blood flow occurring in the usual flare area after HS and SP were almost completely abolished by cetirizine. Inhibition of HS- and SP-induced weal and flare reactions was less marked after loratadine and blood flow in the expanding flare after HS and SP showed significant fluctuations over time. In view of the present results and of data obtained in previous experiments with intradermal injection of agonists, we hypothesize that mode of administration of agonists significantly influences the size of the residual weal after anti-H1 agents. We demonstrate that SP weals induced by pricking are largely inhibited by a potent H1 blockade which supports the view that this phenomenon, as well as the SP-flare, is due to SP-induced histamine liberation. We also, for the first time, report on fluctuations recorded at the edge of the developing flare with laser Doppler flowmetry early after prick testing with a weak H1 blockade. This opens up new avenues in dynamically testing H1-receptor occupancy in vivo and in situ in human skin.


Influence of age and gamma irradiation on the proliferative activity in regenerating rat liver. The effect of aging (2-14 months) and total body irradiation (5.7 Gy of gamma radiation) on liver regeneration was investigated in rats 30 h after partial hepatectomy. Exposure of rats to irradiation 30 min before partial hepatectomy caused latent injury in the remaining liver cells. During the course of liver regeneration this became manifested as a delay in increasing the nucleic acid concentration and content and liver weight and, furthermore, as inhibition of the increase in the mitotic index and cellularity and pronounced increase in the frequency of chromosome aberrations in the postmetaphase. The pattern of age-related changes during liver regeneration was the same as that after irradiation, so that the differences between irradiated and nonirradiated animals became smaller with age.


The use of octadecyl-bonded microparticulate silica in the separation of free and bound fractions during saturation analysis of vitamin D metabolites. The use of octadecyl-bonded microparticulate silica to separate free and bound fractions during the saturation analysis of 25-hydroxyvitamin D and 1,25-dihydroxyvitamin D has been investigated. A slurry of octadecyl-bonded silica in an appropriate incubation buffer was prepared and used in parallel with a conventional dextran-coated charcoal suspension in several assay procedures. Standard curves, non-specific binding and plasma values were compared. A competitive protein binding assay for 25-hydroxyvitamin D and two radioreceptor assays and one radioimmunoassay for 1,25-dihydroxyvitamin D were investigated. In most cases the octadecyl-bonded silica preparation gave the more favourable results; its action was rapid, time- and temperature-independent, and it produced low non-specific binding and higher B0 values in all the assays examined. It was in our hands easier to use than dextran-coated charcoal. The use of octadecyl-bonded silica is recommended as an efficient agent for the separation of free and bound fractions in the saturation analysis of vitamin D metabolites.


Anticancer strategies involving the vasculature: vascular targeting and the inhibition of angiogenesis. Progress in our understanding of tumour angiogenesis and an appreciation of the marked differences between quiescent and proliferating endothelium are revealing new strategies for anticancer therapy. This article outlines these strategies and critically assesses their chances of success.


Activation of platelet concentrate during preparation and storage. This review will discuss how stored platelets become activated and will examine their ability to function and survive in vivo, posttransfusion. Experimental methods which have been shown to alter platelets during storage will be detailed. Using beta-thromboglobulin (beta-TG) and surface adhesion receptors as markers, investigators have examined the activation changes in platelet concentrates during preparation and storage. Resuspension of the platelet pellet after isolation of platelet-rich plasma appears to play a major role in producing platelet activation and beta-TG release during preparation. However, there is a significant amount of interdonor variability in platelet activation even at this early stage of storage. Over 5 days of storage, platelets release approximately 50% of their beta-TG contents. Furthermore, between 40% and 60% of the platelets express the alpha-granule membrane protein, P-selectin (GMP-140), during storage, which is also indicative of platelet activation. These activation changes correlate to some degree with platelet recovery posttransfusion but clearly do not explain the full lesion of platelet storage. The surface density of two platelet membrane receptors, glycoproteins (GP) Ib and IIb/IIIa, also change with activation, although in opposite directions. Platelet surface GPIb decreases initially with storage and then recovers, perhaps due to its relocation to the platelet surface from an intracellular pool. In contrast to GPIb, mean platelet surface GPIIb/IIIa increases slightly during storage, probably as a consequence of platelet activation and release of alpha-granule GPIIb/IIIa to the surface. Some hypotheses are offered regarding how these activated platelets can continue to circulate after transfusion. Further exploration of the platelet storage lesion will hopefully provide needed answers and thus permit better treatment of hemostatic disorders in the future.


Changes in platelet membrane glycoproteins during blood bank storage. The literature on membrane glycoprotein changes occurring during platelet storage is reviewed. Technical problems which may have biased reports are clarified, and the consensus of current studies is established. The membrane alterations that occur during blood bank storage are reminiscent of the surface changes which result from platelet secretion: an increased surface concentration of GP IIb-IIIa and GMP-140, and the appearance of platelet membrane microparticles in the supernatant plasma. The clinical importance of these changes is unknown. Future directions for research on the structure and function of stored platelets are discussed.


Matrix-assisted laser desorption/ionization mass spectrometry of nucleic acids with wavelengths in the ultraviolet and infrared. A number of different matrices have been tested and compared for ultraviolet and infrared (UV and IR) matrix-assisted laser desorption/ionization (MALDI) of oligodeoxyribonucleotides and mixtures thereof, as well as ribonucleic acids (tRNA from yeast and rRNA from E. coli). A new technique for removing alkali cations from nucleic acid samples during sample preparation on the sample support is demonstrated. The amount of oligonucleotide sample consumed during a typical measurement in IR-MALDI-MS was determined.


Temporal differentiation and migration of substance P, serotonin, and secretin immunoreactive enteroendocrine cells in the mouse proximal small intestine. Precise spatial interrelationships exist between substance P, serotonin, and secretin containing enteroendocrine cells in the gastrointestinal tract of mice. In the proximal small intestine these products are coexpressed in various combinations in single enteroendocrine cells along the crypt to villus axis in a pattern that suggests the sequential expression of substance P, serotonin, and secretin. In this report we use bromodeoxyuridine (BrdU) and multilabeling immunohistochemistry to define the temporal and spatial interrelationships between substance P, serotonin, and secretin immunoreactive cells in the mouse proximal small intestine. Our findings demonstrate the sequential expression of substance P, serotonin, and secretin in a population of upwardly migrating enteroendocrine cells and, furthermore, identify a population of crypt associated cells coexpressing substance P and serotonin that fails to traverse this pathway. The lack of secretin immunoreactive cells in the crypts suggests that local factors present in the crypts and/or on villi regulate secretin expression. The combined use of BrdU and multilabeling immunohistochemistry provides a method for defining enteroendocrine cell differentiation pathways throughout the gastrointestinal tract.


[Clinico-epidemiological analysis of the cases of HIV infection in the north-western region of Russia]. The paper provides an epidemiological characterization of HIV infection spread in a Russia's large region with more than 10 million people. The epidemiological findings show that the significant onset of HIV infection occurred among the population in this region in mid 1988. Homosexuals and bisexuals are prevalent among the HIV-infected, sexual contact is the main mode of HIV transmission. In addition to delivery of HIV infection from foreign countries, there are cases of local transmission. The clinical evidence indicates that most HIV-infected people are asymptomatic. Herpes viruses, Mycobacteria tuberculosis, Toxoplasma and fungi are common among causative agents of AIDS-related infections.


[Clinical manifestations and the problems of classification of HIV infection]. The clinical manifestations and some immunological parameters (CD4 lymphocytes, CD4/CD8 ratio, IgM, IgA, IgG levels, skin test) were examined in 226 adult patients (148 males and 78 females) infected with HIV. These included 58 (26%) asymptomatic patients with seropositive test, 109 (48%) with the only clinical manifestation generalized lymphadenopathy; 54 (24%) with AIDS-related infections, 5 (2%) with AIDS. A subsequent follow-up of 3 months to 3 years demonstrated that AIDS developed in 7 patients, 9 died. The period of infection with HIV and death ranged from 1.5 to 9 years. The signs of cell immunodeficiency were found in 70% of the examinees. Recommendations are given on the classification of HIV infection.


[Hemostatic disorders in patients with HIV infection]. Impaired hemostasis was studied in 50 adult patients with HIV infection. The blood coagulative potential, the number and functional activity of platelets were examined. Platelet aggregation and secretion were shown to change earlier than thrombocytopenia developed and clinical signs of HIV infection appeared. The disturbance in the plasma section of hemostasis are due to concurrent opportunistic diseases and infections.


[Pneumocystis carinii infection in a HIV infection department]. To determine the prevalence of Pneumocystis carinii in the HIV-infection department, a simultaneous survey was made of 33 HIV-infected patients at various stages of the disease, of close relatives that were nursing the patients in the unit (n-7), and of medical staff of the department (n-20). Patients with toxic infections who were on another floor of the same hospital and medical students were examined as a control group. For detection of P. carinii antigen, smears from the deep airways were tested in the immunofluorescence. P. carinii was detected in 87.7% of HIV-infected patients, in 71.4% of their relatives and in 80.0% of the medical staff, in 16.6% of control patients and 27.7% of students. The main type of the infectious process in pneumocystosis is its carriage; 2 patients at a stage of relapses (IIIB) that corresponds to AIDS were recorded as having pneumocystis pneumonia.


[Clinico-experimental study of mental processes in men with HIV infection]. A total of 55 males aged 19.5 to 62 years who were infected with HIV were examined psychopathologically+ and neuropsychologically by A.R. Luria's methods. Twenty-one of these patients were homosexuals. Syphilis was recorded in 34.7% in this group. Lymphadenopathy was the major clinical sign of HIV infection. Symptoms of organic involvements+ of the central nervous system were revealed by a psychopathological method. Neuropsychological studies detected dysfunction of the right cerebral hemisphere, particularly in a group of homosexuals suffering from syphilis.


[Comparative analysis of functional activity of peripheral blood phagocytes in HIV-infected patients and in those with recurrent herpes simplex]. The functional activity of peripheral phagocytes were comparatively studied in 14 HIV-infected patients and 28 patients with chronic Herpes simplex viral infection. The two groups exhibited lowered adhesive capacity of phagocytes, impaired production and excretion of active oxygen metabolites. In addition, the patients with chronic Herpes simplex infection showed much elevated levels of myeloperoxidase and acid phosphatase, which indicated its compensatory pattern. The HIV infected had no enhanced enzymatic activity. One cannot rule out that these differences in the functional activity of phagocytes are associated with different effects of viral peptides on the cellular wall of phagocytes.


[Tumor necrosis factor-alpha and interleukin-1beta in the blood plasma of patients with HIV infection]. The paper presents examination findings of 67 HIV-infected patients: 32 children aged 2.5 to 16 years and 35 adults aged 21 to 46 years. The proportion of patients with higher alpha-tumor necrosis factor and interleukin-1 beta increased with progression of the disease, the two parameters being higher in the children than in the adults. A correlation was found between the appearance of cytokines in the plasma and some clinical signs, such as fever, weight loss, anemia, neurological disorders. A parallel study of the patients' immunograms indicated significant correlations between the degree of impairments in the composition of lymphocyte subpopulations and the appearance of plasma cytokines.


[Detection of Mycoplasma and its antibodies in the blood of HIV-positive patients and in healthy persons]. The comparative study of HIV-positive and clinically healthy persons has indicated that the antigens of M. pneumoniae, M. fermentans and U. urealyticum are encountered nearly twice more frequently in the blood of HIV-infected patients than in that of healthy individuals. Mycoplasma antibodies are detected in HIV-positive persons 12 times more frequently than in healthy ones. Among the HIV-infected persons there are those who have simultaneously antigens of some Mycoplasma species.


[The immunoenzyme test system for detection of HIV-1 antigens based on using immune polyclonal anti-HIV serum and monoclonal antibodies against gene GAG HIV-1 proteins]. The paper describes the enzyme immunoassay system for detection of human immunodeficiency virus antigens, which is based on the use of rabbit anti-HIV antibodies and monoclonal antibodies to HIV-1 gene proteins gag. The system may be useful in the examination of laboratory and clinical samples to reveal both free and conjugated antigens in the composition of immune complexes. The sensitivity of the assay system under development is 0.5 ng/ml at 100% specificity.


[Peptides from the principal neutralizing and CD4-binding domain: similar immunoreactive properties and structure pattern]. The human immunodeficiency virus (HIV) proteins gp120 and gp41 are the principal immune target in HIV infection. One of the most important trends in the study of AIDS is linked to the mapping of sites involving in the binding to the cell receptor CD4 and in the induction of virus-neutralizing antibodies (VNA). Recent studies have revealed that gp120 as the major domain contains inducing type-specific BNA (PND) and a binding region with CD4 (CD4-BR). PND is located in the hypervariable loop of gp120 (residues 301-336 for a BRU strain), and CD4-BR is in the conservation area (residues 410-450). By using the synthetic fragments from these areas (BRU and MN strains) and HIV-infected persons' sera, the authors established that the immune response to PND and CD4-BR is somewhat interrelated: there is a synchronized response of HIV antibodies to peptides from the two regions in ELISA (r = 0.82). For analysis of this phenomenon, experiments with cross-linked immunoreactivity of rabbit antisera to peptides from PND and CD4-BR with homologous and heterologous peptides were performed by applying three control peptides from HIV and hepatitis B virus. It has been found that there is a cross reactivity between rabbit anti-PND (MN, BRU) and anti-CD4-BR abs. Peptide homological analysis revealed common structural elements for PND and CD4-BR despite significant differences in their proposed functions. There is a large amount of positively charged aa within both PND and CD4-BR which may be involved in gp120-CD4 interaction. Acetylation of Lys residues resulted in complete loss of peptide reactivity.


[Expression of HIV-1 tat gene under the control of P 7,5 KD vaccinia virus promoter in CV-1 cells]. The transient expression of the HIV-1 gag genes and a HIV-1 ++trans-activator protein (tat)-encoded was made in cultured CV-1 cells. In recombinant plasmids, the gag gene was under the control of HIV-1 ++trans-activator sequence (tar) and the tat gene was under the control of a 7.5-kd vaccinia promoter. Transactivation of gag gene expression, which was stimulated by a tat gene expression product, was observed in the presence of wild vaccinia virus. The transaction was immunologically evaluated from the binding to monoclonal anti-p17 and anti-p24 antibodies. The findings lead to the discussion whether the regulatory proteins of HIV-1 can express in vaccinia virus vectors.


[Study of antigenic structure of HIV-1 protein p24 using monoclonal antibodies]. During the experiments 4 murine and 3 rat hybridomas producing monoclonal antibodies (MAb) against the protein p24 of human immunodeficiency virus type 1 (HIV-1) have been obtained. Using the immunoblotting technique, it was established that all the species of MAb reacted with the same viral proteins which are derivatives of gag gene--p24 and p55. The properties of MAb have been studied in competitive binding. Their ability of binding to different fragments of the gag protein produced by the recombinant plasmids in E. coli cells have been investigated in ELISA. The analysis of the findings suggests that the HIV-1 protein p24 contains at least 3 antigenic epitopes. All species of MAb reacted with 3 different HIV-1 strains and 2 HIV-1 isolates, but failed with 2 different HIV-2 strains. The only MAb NS5E4 can be used as an immunosorbent in the antigenic capture reaction.


[Persistent viruses in serological diagnosis of HIV infection]. Specific antibodies to persistent viruses (CMV, EBV, HBV) were detected by ELISA in groups of HIV-infected patients and persons showing indefinite results of the immunoblotting test for HIV-1 antigens, on the one hand, and in HIV-seronegative donors and patients with clinical manifestations of viral infection (CMVI) on the other. The findings indicate that the persons with indefinite immunoblotting test results show elevated blood CMV and HBV antigen levels than in the matched group of seronegative donors. This fact suggests that persistent viral infections might involve in the formation of an indefinite pattern when the sera were tested for HIV. The patients whose sera behaved in such a way represent a clinical risk group for HIV infection and call for further follow-up.


The effects of purified 25-kDa lipase from a clinical isolate of Pseudomonas cepacia in the lungs of rats. Nanogram quantities of a 25-kDa lipase purified from culture supernatants of Pseudomonas cepacia 90ee, a sputum isolate from a cystic fibrosis (CF) patient, were placed in the lungs of healthy rats. The resulting pathological changes included large amounts of proteinaceous exudate, the accumulation of polymorphonuclear leukocytes and red blood cells, and disorganization of alveolar structure. Pseudomonas cepacia 90ee immobilized in agar beads was also placed in the lungs of rats in a model of chronic infection. This resulted in bronchopneumonia and a milder inflammatory response than that elicited by the purified enzyme.


Electrophoretic analysis of heterogeneous lipopolysaccharides from various strains of Vibrio vulnificus biotypes 1 and 2 by silver staining and immunoblotting. Lipopolysaccharides (LPS) of 11 strains of Vibrio vulnificus biotypes 1 and 2, isolated from an eel farm, and of 10 reference strains, were examined by SDS-polyacrylamide gel electrophoresis coupled with silver staining and immunoblotting. LPS samples were obtained from whole-cell lysates, outer membrane fragments, and extracellular products. By silver staining, only a diffuse band of low-molecular weight could be visualized in all cases except for a biotype 1 strain isolated from water. However, immunoblotting with antisera obtained against strains of biotypes 1 and 2 from eels allowed visualization of multiple O-polysaccharide chains. All biotype 2 strains, independently of their origins, belonged to the same serotype and presented the same LPS profile, whereas eel isolates of biotype 1 were serologically identical and different from the rest of tested strains of biotype 1. This is the first report of LPSs with a ladder-like structure in Vibrio vulnificus.


Differentiation between parasystole and reentry in concealed bigeminy. There are two different theories to explain the mechanism of concealed bigeminy: one is '2:1 concealed reentry'; the other is 'irregular parasystole.' Two exemplary cases of the even-number variant of concealed bigeminy are presented. In case 1, the mechanism can be explained by an irregular parasystole due to a modulated parasystole; however, findings during temporary sinus arrest caused by vagal stimulation indicate that this case is not governed by a parasystole, but by a 2:1 concealed reentry. In case 2, the mechanism can be explained by a 2:1 concealed reentry without parasystole; however, findings during temporary sinus arrest indicate that this case is governed by an irregular parasystole due to a type-I second-degree entrance block. Thus, in cases of concealed bigeminy without pure ectopic cycles, it does not seem easy to explain the mechanism of concealed bigeminy on the theory of a modulated parasystole.


Administration of human recombinant insulin-like growth factor-I to patients following major gastrointestinal surgery. The aim was to study the pharmacokinetic parameters and biological activity of a single dose of human recombinant IGF-I (rhIGF-I) administered to patients following major gastrointestinal surgery. A double blind placebo controlled externally randomized study of 30 patients; the study commencing 24 hours after major colonic or gastric surgery. After a baseline blood sampling day, IGF-I (40 micrograms/kg by single subcutaneous dose, n = 20) or placebo (n = 10) was administered and serum and urine samples collected over the ensuing 72 hours. Serum IGF-I, IGF-II, IGF binding proteins (IGFBP-1, IGFBP-3), GH and insulin were measured by radioimmunoassay. Serum IGF bioactivity was assessed using a validated porcine cartilage bioassay. Serum and urinary electrolytes were measured by standard methodology. Serum immunoreactive IGF-I levels peaked at 4 hours following injection of IGF-I (1.09 +/- 0.12 U/ml mean +/- SEM), remained elevated for 15 hours and returned to basal levels by 24 hours after injection. IGF bioactivity was increased by 57% 6 hours after IGF-I injection. Mean levels of IGFBP-1 and IGFBP-3, IGF-II and GH were unaffected by IGF-I administration. Insulin levels were suppressed at 30 minutes following injection of IGF-I compared with the placebo group (16.9 +/- 3.0 mU/I vs 32.3 +/- 7.1, P = 0.02); thereafter, there were no differences in insulin levels. The mean change in serum creatinine following IGF-I (-6.3 +/- 3.0 mmol/l) was significantly different from that in the control group (+7.2 +/- 6.2, P = 0.03). Creatinine clearance rose from a mean of 71.6 +/- 7.5 ml/min to 83.2 +/- 7.6 ml/min after IGF-I treatment (P = 0.02). In the IGF treated patients, cholesterol levels consistently fell (-0.20 +/- 0.05 mmol/l); this was not observed in the placebo group (+0.20 +/- 0.14, P = 0.006). Basal serum potassium levels in the IGF treatment group (4.1 +/- 0.1 mmol/l) fell to 3.8 +/- 0.1 at 4 hours (P = 0.002) and 3.6 +/- 0.1 at 10 hours (P = 0.001) returning to a level of 4.0 +/- 0.1 (P = 0.293) at 24 hours after injection. There were no other observed differences in serum or urinary electrolytes or serum free fatty acids and triglycerides. Pharmacokinetic parameters derived from baseline adjusted IGF-I measurements revealed a slow absorption of the administered dose with a Tmax of 5.0 +/- 0.43 hours and an elimination half-life of 10.8 +/- 1.2 hours. The computed volume of distribution was 0.33 +/- 0.05 I/kg and the clearance on average 25 ml/min. A single subcutaneous dose of IGF-I normalized circulating IGF-I levels in post-operative patients, was well tolerated and without side-effects. IGF bioactivity was increased and associated with a fall in serum cholesterol, potassium and creatinine levels and a rise in creatinine clearance. Further long-term studies are now required to assess the anabolic effects of rhIGF-I in this type of patient group.


Galanin inhibits tolbutamide-stimulated insulin secretion in the perfused pig pancreas. The neuropeptide galanin is known to inhibit insulin secretion in a variety of species. However, controversies exist regarding its action on insulin secretion in the perfused pig pancreas, since both inhibitory and stimulatory effects have been described. We therefore perfused the isolated porcine pancreatico-duodenal block with galanin (100 nmol/l) in the presence of 5 or 15 mmol/l glucose or with 5 mmol/l glucose and 10 mumol/l tolbutamide. We found that at this dose level, galanin did not affect insulin secretion stimulated by glucose alone. In contrast, galanin clearly suppressed tolbutamide-stimulated insulin secretion. Hence, we conclude that galanin has a weak influence on insulin secretion in the pig pancreas, being unable to inhibit glucose-stimulated insulin secretion at the dose level of 100 nmol/l. However, when active, galanin clearly inhibits insulin secretion also in the pig pancreas.


Comparison of top and bottom loading of a dextran gradient for rat pancreatic islet purification. Rat pancreatic islet yields obtained with dextran gradient purification were compared after suspending the digest into either the top or the bottom layer of the gradient. A 5-layer discontinuous gradient was used, which consisted of 16 ml 31% dextran as bottom layer, overlayered with 25%, 23%, 20% and 11% dextran (4 ml each). When the digest of 1 rat pancreas was suspended into the top layer of the gradient, the total number of islets obtained from the 11-20, 20-23 and 23-25% interfaces was 862 +/- 38, 240 +/- 39 and 54 +/- 5, respectively. From this gradient, also 1409 +/- 81 islets were retrieved from the bottom layer (i.e., exocrine pellet). In contrast, when the pancreas digest was suspended into the bottom layer of the gradient, 1964 +/- 63, 435 +/- 42, and 177 +/- 34 islets were obtained from the successive interfaces, and only 50 +/- 20 islets from the exocrine pellet. The total islet volume obtained from the two uppermost interfaces was 3.46 +/- 0.31 microliters after top-loading, and 4.93 +/- 0.16 microliters after bottom-loading (n = 7, p < 0.01). When the islets retrieved from one bottom loaded gradient were transplanted into either 1 (n = 6) or 2 (n = 9) diabetic recipients, glucose levels normalized in all instances. We therefore conclude that a bottom-loaded dextran gradient separates islets from exocrine tissue effectively, resulting in significantly higher islet yields than obtained with a top-loaded dextran gradient.


An ultrastructural study of the effects of topical tretinoin on microcomedones. It has been hypothesized that topical tretinoin prevents inflammatory acne lesions by loosening follicular impactions (microcomedones) and clearing the follicular canal of retained keratin. To lend support to this hypothesis, 15 volunteers applied 0.1% tretinoin cream once daily for 12 weeks to one side of the face and an emollient cream to the other side. During the 12-week treatment period, samples of microcomedones were obtained using the follicular biopsy technique. By 6 weeks, the number of microcomedones was reduced approximately 50% from baseline on the side treated with tretinoin and approximately 80% at the end of the study. Only a minimal reduction in the number of microcomedones was seen on the emollient side, even after 12 weeks of application. Morphologic examination showed a progressive loss of the cohesiveness of the microcomedones and ultrastructural alterations in the epithelial cells treated with tretinoin. The microcomedones changed from well-developed keratinous plugs containing many bacteria to a few wispy layers of keratin containing few bacteria. This study confirms the microcomedolytic activity of tretinoin and provides evidence that this activity is associated with changes in the differentiation of the follicular epithelial cells.


Antibacterial properties of investigational, new, and commonly used antibiotics against isolates of Pseudomonas cepacia in Michigan. Microdilution antimicrobial susceptibility testing was performed with 73 isolates of Pseudomonas cepacia collected from the sputum of patients throughout Michigan with cystic fibrosis. Susceptibility testing was done using new and investigational antibiotics (loracarbef, cefixime, cefpirome, desacetyl-cefotaxime, cefpodoxime, cefmetazole, cefepime, cefprozil, and fleroxacin) and commonly used antibiotics (ceftazidime, mezlocillin, piperacillin, ciprofloxacin, tobramycin, and amikacin). Ceftazidime was the most active antibiotic, and 91.8% of isolates were susceptible to it with MIC50 and MIC90 values of < or = 4 and 16 micrograms/ml, respectively. For mezlocillin, piperacillin, and ciprofloxacin 84.9, 89 and 39.7% of the isolates, respectively, were mostly moderately susceptible. Loracarbef, cefixime, cefprozil, cefmetazole, cefepime, fleroxacin, cefpodoxime, tobramycin, and amikacin did not show activity against P. cepacia. For cefpirome and desacetylcefotaxime 24.7 and 60.3% of the isolates, respectively, were moderately susceptible. Both MIC50 and MIC90 were > 32 micrograms/ml for cefpirome and 32 and > 64 micrograms/ml for desacetylcefotaxime.


Vital dye analysis of cranial neural crest cell migration in the mouse embryo. The spatial and temporal aspects of cranial neural crest cell migration in the mouse are poorly understood because of technical limitations. No reliable cell markers are available and vital staining of embryos in culture has had limited success because they develop normally for only 24 hours. Here, we circumvent these problems by combining vital dye labelling with exo utero embryological techniques. To define better the nature of cranial neural crest cell migration in the mouse embryo, premigratory cranial neural crest cells were labelled by injecting DiI into the amniotic cavity on embryonic day 8. Embryos, allowed to develop an additional 1 to 5 days exo utero in the mother before analysis, showed distinct and characteristic patterns of cranial neural crest cell migration at the different axial levels. Neural crest cells arising at the level of the forebrain migrated ventrally in a contiguous stream through the mesenchyme between the eye and the diencephalon. In the region of the midbrain, the cells migrated ventrolaterally as dispersed cells through the mesenchyme bordered by the lateral surface of the mesencephalon and the ectoderm. At the level of the hindbrain, neural crest cells migrated ventrolaterally in three subectodermal streams that were segmentally distributed. Each stream extended from the dorsal portion of the neural tube into the distal portion of the adjacent branchial arch. The order in which cranial neural crest cells populate their derivatives was determined by labelling embryos at different stages of development. Cranial neural crest cells populated their derivatives in a ventral-to-dorsal order, similar to the pattern observed at trunk levels. In order to confirm and extend the findings obtained with exo utero embryos, DiI (1,1-dioctadecyl-3,3,3',3'-tetramethylindo-carbocyanine perchlorate) was applied focally to the neural folds of embryos, which were then cultured for 24 hours. Because the culture technique permitted increased control of the timing and location of the DiI injection, it was possible to determine the duration of cranial neural crest cell emigration from the neural tube. Cranial neural crest cell emigration from the neural folds was completed by the 11-somite stage in the region of the rostral hindbrain, the 14-somite stage in the regions of the midbrain and caudal hindbrain and not until the 16-somite stage in the region of the forebrain. At each level, the time between the earliest and latest neural crest cells to emigrate from the neural tube appeared to be 9 hours.(ABSTRACT TRUNCATED AT 400 WORDS)


Requirement of c-kit for development of intestinal pacemaker system. A discovery that the protooncogene encoding the receptor tyrosine kinase, c-kit, is allelic with the Dominant white spotting (W) locus establishes that c-kit plays a functional role in the development of three cell lineages, melanocyte, germ cell, and hematopoietic cell which are defective in W mutant mice. Recent analyses of c-kit expression in various tissues of mouse, however, have demonstrated that c-kit is expressed in more diverse tissues which are phenotypically normal in W mutant mice. Thus, whether or not c-kit expressed outside the three known cell lineages plays a functional role is one of the important questions needing answering in order to fully elucidate the role of c-kit in the development of the mouse. Here, we report that some of the cells in smooth muscle layers of developing intestine express c-kit. Blockade of its function for a few days postnatally by an antagonistic anti-c-kit monoclonal antibody (mAb) results in a severe anomaly of gut movement, which in BALB/c mice produces a lethal paralytic ileus. Physiological analysis indicates that the mechanisms required for the autonomic pacing of contraction in an isolated gut segment are defective in the anti-c-kit mAb-treated mice, W/Wv mice and even W/+ mice. These findings suggest that c-kit plays a crucial role in the development of a component of the pacemaker system that is required for the generation of autonomic gut motility.


Caprine beta-mannosidosis: regional differences in deficits of CNS myelin proteins. Caprine beta-mannosidosis is an autosomal recessive disorder characterized by marked deficiency of beta-mannosidase activity, accumulation of oligosaccharides, and pathologic changes involving prominent dysmyelination. The myelin deficits show marked regional variation, with spinal cord mildly affected and many brain regions severely affected by morphologic criteria. In this study, levels of myelin basic protein (MBP) and proteolipid protein (PLP) were measured by immunoblotting in samples prepared from spinal cord, brainstem and cerebral hemispheres of normal and affected goats at 2-4 days (newborns) and 2-4 weeks of age. In affected goats, total levels of MBP in spinal cord were normal, while PLP levels were 60-70% of normal at both ages. In contrast, PLP and MBP in brainstem and cerebral hemispheres were severely decreased at both ages, with levels of PLP 10-13% and MBP 25-29% of normal in newborns, and generally more reduced at 2-4 weeks. When myelin fractions were isolated on 0.32/0.85 M sucrose gradients, yields were about 38 and 25% of normal in spinal cord at the two ages, but less then 3% of normal in brainstem. Yields of myelin-like fraction were decreased as well, but to lesser extents than yields of myelin. Myelin from spinal cord had a normal composition with regard to PLP and MBP content, while the myelin fraction from brainstem was markedly deficient in both proteins. This suggests formation of myelin with a very abnormal composition in brainstem, or inclusion of large amounts of membranes other than myelin in this fraction. The more severe deficits in brainstem and cerebral hemispheres compared to spinal cord are consistent with morphologic observations.(ABSTRACT TRUNCATED AT 250 WORDS)


A rat monoclonal antibody specific for murine type 1 pneumocytes. A rat monoclonal antibody (MAb), 411-52, that binds specifically to murine pulmonary alveolar type 1 cells was developed. The cell-binding specificity of MAb 411-52 was assessed by light microscopy on immunoperoxidase-labeled tissue sections, electron microscopy on immunogold-labeled tissue blocks, and by flow cytometric analysis and fluorescence-activated cell sorting of immunofluorescently labeled cells enzymatically dissociated from murine lungs. The epitope recognized by MAb 411-52 was first detected in immunoperoxidase-stained sections of neonatal lungs of mice approximately 3 weeks after birth. In adult mice, the MAb 411-52-directed, immunoperoxidase-staining pattern was uniform throughout the lung parenchyma, was restricted to the luminal surfaces of alveoli, and was absent from type 2, endothelial, and interstitial cells, as well as from the epithelial cells of conducting airways. Electron microscopic analysis of immunogold-labeled lung tissue confirmed the type 1 cell binding specificity of MAb 411-52. Analysis by multiparameter, laser flow cytometry indicated that MAb 411-52 binds to 4.6 +/- 0.5% (mean +/- SD) of enzymatically dissociated cells from the lungs of normal adult mice. The absence of immunogold-labeling of type 2 cells suggested that the epitope recognized by MAb 411-52 might be a differentiation marker for the type 1 cell phenotype. With this MAb and standard immunohistochemical techniques, it is possible to visualize directly type 1 cells in paraffin sections.


The contrast sensitivity function in low vision. The contrast sensitivity of 51 low vision patients (95 eyes) showed a substantial decrease in all spatial frequencies. The peak contrast sensitivity was shifted to 1 C/D from the normal 3-4 C/D. The high-frequency cut off is correlated to the visual acuity positively. Of the 33 patients, the preferred eye was the eye with better peak sensitivity in 28 patients (84%), while the preferred eye was the eye with better visual acuity in 22 patients (66%). It seems that the peak sensitivity is more important than visual acuity in determining eye preference. It provides a useful information for the clinician to determine which eye to train with low vision aids. Forty patients among 46 patients (87%) with RFN (Recognized Frequency Number) > or = 3 are able to read No.5 reading card, while 5 patients among 5 patients with RFN < 3 are not able to read No.5 reading card. The contributions of CSF to the low vision are evaluated.


Cloning of a type I keratin from goldfish optic nerve: differential expression of keratins during regeneration. We report the cDNA sequence and predicted amino acid sequence of a novel type I keratin, designated as GK50, and show that keratin expression in the goldfish optic nerve is highly complex. The GK50 protein is one of at least three type I keratins expressed in goldfish optic nerve based on both antibody reactivity and blot-binding to the type II keratin ON3. After optic nerve crush in situ hybridization shows a localized increase in GK50 mRNA expression in the crush zone. This is in contrast to ON3 mRNA which shows a localized increase that is limited to the proximal and distal margins of the crush zone, suggesting a diversity of keratin expression in different cell types of the goldfish optic nerve.


Cytokeratin 14 expression in rat liver cells in culture and localization in vivo. Rat liver epithelial cells (LECs) are non-parenchymal proliferating cells that readily emerge in primary culture and can be established as cell lines, but their in vivo cell(s) of origin is unclear. We reported recently some evidence indicating that the LEC line, T51B, contains two cytokeratins (CKs) equivalent to human CK8 and CK14 respectively. T51B cells also contain vimentin assembled as a network of intermediate filaments distinct from that of the CKs. In the present study, we examined the expression of CK14 gene in various LEC preparations and a Triton-resistant rat skin cytoskeletal fraction, and then assessed its usefulness as an LEC specific marker in the liver. Northern and Western blot analyses with cDNAs and antibodies for CK8, CK14, CK18 and vimentin confirmed that rat hepatocytes express CK8 and CK18 genes only, whereas T51B cells express CK8, CK14 and vimentin genes in the absence of CK18. CK14 was also present in LECs derived as primary from embryonic-day 12 rat liver and secondary cultures from 4-day-old rat liver. Primary cultures of oval cells isolated from 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB) treated rat liver (an enriched source of biliary epithelial cells) contained CK14 mRNAs which were slightly shorter than those in LECs. The analyses of CK5 (the usual partner of CK14) gene expression using specific cDNA and antibody clearly demonstrated its absence in LECs. In situ double immunolocalization analyses by laser scanning confocal microscopy showed that CK14 was not present in hepatocytes (HES6+ cells) and was expressed in some biliary epithelial (BDS7+ cells). CK14-positive cells were also found in the Glisson's capsule. However, CK14-positive cells of the portal region were vimentin negative, whereas those of the Glisson's capsule were vimentin positive. Our results suggest that CK14 gene expression is part of the differentiation program of two types of LECs and that this differential CK14 gene expression can be used as a new means to type LECs in culture and in vivo.


Characterization of an endothelial protein in the developing rabbit kidney. A new protein (EnPo 1 antigen) abundant on endothelial cells and glomerular podocytes has been characterized by means of the mouse monoclonal antibody EnPo 1. Following electrophoretical separation of rabbit kidney homogenates EnPo 1 recognized a protein with a molecular weight of 110 kDa and an isoelectric point of 5.9 in Western blots. Using immunohistological techniques, the EnPo 1 antigen has been localized in high concentrations on glomerular podocytes of different developmental stages. Furthermore, the EnPo 1 antigen was expressed on endothelial cells of all adult rabbit organs tested so far. Detailed analysis of neonatal rabbit kidney revealed the abundance of EnPo 1 antigen on both differentiated vessels as well as on immature endothelial cells and endothelium of the microvasculature. Thus, for the first time a marker for in situ investigations of angiogenic processes within the mammalian kidney is available. Analysis of kidney cryosections by confocal laser scan microscopy revealed a direct connection between mature and differentiating vessels in the outer kidney cortex. Furthermore, two differentially organized cell populations discriminated by their EnPo 1 binding pattern were localized in the embryonic renal cortex. Morphologically, these cells were not distinguishable from other mesenchymal cells.


Modification of the in vitro cytotoxicity of hydrogen peroxide by iron complexes. The effect of a range of iron chelates on the cytotoxicity of H2O2 was studied on a mammalian epithelial cell line. Iron complexes which were internalised enhanced the cytotoxicity of H2O2 measured by delayed thymidine incorporation. Iron complexed to 8-hydroxyquinoline (Fe/8-HQ) potentiated the cytotoxicity of 50 microM by 38% and Fe/dextran by 23%. Pre-exposure of cells to Fe/dextran at 4 degrees C did not result in any potentiation of H2O2-induced cytotoxicity which we ascribe to failure of the Fe/dextran to be endocytosed at low temperature. Iron complexes which are slowly taken up or remain extracellular protected the cells from H2O2-induced cytotoxicity. Thus, Fe/EDTA inhibited the cytotoxicity of 50 microM H2O2 by 33%; Fe/ADP by 80% and Fe/ATP by 88%, suggesting mutual extracellular detoxification.


Conformational study of a peptide epitope shows large preferences for beta-turn conformations. The conformational preferences of the 7-residue peptide Glu-Val-Val-Pro-His-Lys-Lys was investigated using a global search algorithm, namely the Electrostatically Driven Monte Carlo (EDMC) method, and the ECEPP/2 potential energy function. This particular sequence corresponds to the N-terminal portion of a 19-residue peptide antigen whose three dimensional structure, when complexed to a cognate antibody, was reported recently. As a result of this study a series of low-energy conformations were identified showing a common folding pattern with residues Val-3, Pro-4, His-5 and Lys-6 forming a beta turn. A comparison of the computed conformations with the one determined by X-ray crystallography in the antibody-antigen complex reveals marked similarities. In most of the cases rms deviations smaller than 1.1 A were found for the backbone atoms of the four residues forming the turn. These results suggest that the recognition process is accomplished in this case through the interaction of the antibody with relatively stable conformers of the antigenic peptide.


DP epitope mapping by using T-cell clones. To determine whether a correlation exists between the genomic HLA class II DP DNA polymorphism and cell surface expression and to detect the DP epitopes responsible for alloreactivity, anti-DP T-cell clones were generated against new PLT blank RFLP DPa and DPb-defined specificities. The clones were tested on the 10th IHWS B-LCLs and on local panel cells. Oligotyping of the tested cells made it possible to (a) correlate the DPa specificity with the DPB1*0402 specificity and (b) split DPb into DPB1*1001 and DPB1*1401. By comparing DNA sequences of the second exon to panel reactivity, the epitopes responsible for DPB1*1001 and 1401 were defined and attributed to beta-chain residues contributing to peptide selection inside the HLA groove. However, DNA sequences could not explain anti-DPa allospecificity, indicating that another structure not yet definable may be involved.


Mutagenesis around residue 176 on HLA-B*0702 characterizes multiple distinct epitopes for anti-HLA antibodies. Preformed antibodies against HLA-A,B,C molecules cause hyperacute rejection of transplanted allogeneic tissues. To understand better the molecular basis of hyperacute rejection, narrowly reactive anti-HLA-B*0702 monoclonal antibodies have been studied. Previous epitope mapping studies of these monoclonal antibodies by mutating B*0702 have conflicted with antibody-blocking studies. To resolve these discrepancies, we mutated B*0702 residues around the antigenically important residue 176, and measured anti-B*0702 antibody binding. Antibody MB40.2 binding is abrogated by mutations at residues 169, 180, and 182, close to residue 176 in the primary structure. However, MB40.2 binding is not affected by 12 other B*0702 mutations close to residue 176 in the tertiary structure. This suggests that MB40.2 may recognize a sequential B*0702 epitope including residues between positions 169-182. Antibody BB7.1 binding requires B*0702 alpha 2-domain residues 166 and 169. Competition for B*0702 residue 169 explains why MB40.2 and BB7.1 crossblock. Because BB7.1 binding also requires B*0702 alpha 1-domain residues, BB7.1 may contact both alpha-helices, straddling the B*0702 peptide-binding groove. Previous results showed that both MB40.2 and MB40.3 binding require B*0702 residues 176/178. However, MB40.3 binding is not affected by any of 15 other mutations near residue 176. This suggests that MB40.3 does not contact residues 176/178; rather, residues 176/178 appear to affect MB40.3 binding by subtly influencing B*0702 conformation. Thus, monoclonal antibodies influenced by a defined B*0702 region around residue 176 appear to recognize three different types of epitopes. This suggests that human alloantibodies also recognize diverse types of HLA epitopes.