Distinct N-glycan fucosylation potentials of three lepidopteran cell lines. The fucosyltransferase activities of three insect cell lines, MB-0503 (from Mamestra brassicae), BM-N (from Bombyx mori) and Sf-9 (from Spodoptera frugiperda), were investigated and compared with that of honeybee venom glands. Cell extracts and venom gland extracts were incubated with GDP-[14C]fucose and glycopeptides isolated from human IgG and from bovine fibrin. The labeled oligosaccharide products were released by peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase A, fluorescence marked with 2-aminopyridine and analyzed both by reversed-phase and size-fractionation HPLC. They were identified by their elution positions before and after exoglycosidase treatment in comparison with standard oligosaccharides. These experiments revealed distinct fucosylation potentials in the three cell lines tested. While MB-0503 cells, like honeybee venom glands, are able to transfer fucose into alpha 1-3 and alpha 1-6 linkage to the innermost N-acetylglucosamine, only alpha 1-6-fucosyl linkages were detected with BM-N and Sf-9 cells.


Primary structure of neutral and acidic oligosaccharide-alditols derived from the jelly coat of the Mexican axolotl. Occurrence of oligosaccharides with fucosyl(alpha 1-3)fucosyl(alpha 1-4)-3-deoxy-D-glycero-D-galacto-nonulosonic acid and galactosyl(alpha 1-4)[fucosyl(alpha 1-2)]galactosyl(beta 1-4)-N-acetylglucosamine sequences. Six major neutral and acidic oligosaccharide-alditols were prepared from the jelly coat of Mexican axolotl eggs. These compounds were demonstrated to contain 3-deoxy-D-glycero-D-galacto-nonulosonic acid (dNloA) and L-fucose (Fuc). The structures of the six major oligosaccharides were established as follows: [sequence: see text]


Molecular hybridization techniques in current diagnosis of chronic hepatitis B in childhood. Following the cloning and sequencing of the hepatitis B virus genome, molecular hybridization techniques have been established to detect hepatitis B virus (HBV) DNA in serum and liver tissue. Analyses can be performed by dot blot, Southern blot and in situ hybridization. HBV DNA is regarded to be the most sensitive marker of viral replication and infectivity which was previously related to the presence of hepatitis B e antigen in serum and hepatitis B core antigen in liver cells. In liver tissue different molecular patterns can be recognized as free viral DNA and integrated sequences. Furthermore, introduction of the polymerase chain reaction allows the detection of very small amounts of viral DNA and has markedly improved diagnostic sensitivity. Thus the study of HBV DNA has become a valuable part of the routine diagnosis in chronic hepatitis B, providing a more reliable evaluation of virus replication and infectivity, and facilitating more precise statements about course and prognosis of the disease.


Transient cardiac constriction following purulent pericarditis. Transient cardiac constriction is an unusual complication of purulent pericarditis. It should be suspected in the presence of clinical and haemodynamic deterioration when signs of activity have abated. Features of cardiac constriction were observed in a 4-year-old boy 2 weeks after surgical drainage. The patient was managed conservatively without surgery and the outcome was good. Follow up 2 years later showed a healthy boy with a normal cardiological examination.


Lethal junctional epidermolysis bullosa with normal expression of BM 600 and antro-pyloric atresia: a new variant of junctional epidermolysis bullosa? A newborn girl is described with a lethal junctional epidermolysis bullosa (Herlitz form) (JEB) associated with a congenital localized absence of skin, and a pyloric atresia (PA). The post-mortem examination of the digestive tract showed a widespread cleavage between the epithelium and the chorion. Immunohistological and electron microscopical examination showed a cleavage occurring through the lamina lucida of the digestive basement membrane, as for the skin blisters. Despite the lethal character of this form of JEB, the BM 600 glycoprotein was normally recognized at the dermo-epidermal junction by the monoclonal antibody GB3. This rare association of lethal JEB-PA-localized absence of skin, with a quite unusual GB3 positive immunophenotype could correspond to a new variant of JEB.


Development of height and weight in children with diabetes mellitus: report on two prospective multicentre studies, one cross-sectional, one longitudinal. Optimal regimen for insulin therapy should lead to normal longitudinal growth and weight gain in children with diabetes mellitus. However, reports published so far indicate that this goal of paediatric diabetology is currently not achieved in a considerable number of patients. In a cross-sectional sample of 89 children with insulin dependent diabetes mellitus (IDDM) for more than 3 years, we found the relation of height to weight to be significantly different compared to 102 healthy school children of similar age. Using bivariate analysis, body shape in these children with diabetes was shifted towards small and obese (P less than 0.05) compared to control children. We subsequently initiated a longitudinal study and followed children from the onset of diabetes for the following 3 years, recording height, weight and bone age as well as glycosylated haemoglobin and daily insulin requirement. At diagnosis, height SDS was identical in children with IDDM (+0.04 +/- 0.10) compared to control children (-0.07 +/- 0.10; M +/- SE), while weight SDS was -0.26 +/- 0.10 in children with diabetes (controls: + 0.01 +/- 0.01). Bone age was identically retarded in newly diagnosed IDDM children (-0.73 +/- 0.12 SDS) and in our control group of children from the same regional background (-0.50 +/- 0.12; n.s.). In this group of children with diabetes mellitus followed prospectively, height to weight relationship differed from controls after 2 and after 3 years of the disease (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Diagnostic value of growth hormone-releasing hormone test in children and adolescents with idiopathic growth hormone deficiency. Average growth hormone (GH) peaks following an i.v. growth hormone releasing hormone (GHRH) 1-29 stimulation test were significantly lower in 48 children and adolescents with GH deficiency (GHD) than in 20 age-matched controls (15.2 + 12.7 vs 37.5 + 28.1 ng/ml, 2 P less than 0.001). Twelve patients exhibited a low GH peak (less than 5 ng/ml), 27 demonstrated a normal response (greater than 10 ng/ml) and 9 showed an intermediate rise in plasma GH (5-10 ng/ml). Six of the 12 patients with low GH response to the first GHRH stimulation failed to respond to two other tests immediately before and after a 1 week priming with s.c. GHRH. These subjects with subnormal GH increase at repeat testing had total GHD (TGHD) and multiple pituitary hormone deficiency (MPHD) and had suffered from perinatal distress. On the contrary, 26 of 27 patients with normal GH response to the first test had isolated GHD and only a minority (8/27) had signs of perinatal distress. It is concluded that perinatal injuries primarily damage pituitary structures and that a pituitary defect more probably underlies more severe forms (TGHD and MPHD) of GHD.


The association of hypopituitarism with small pituitary, invisible pituitary stalk, type 1 Arnold-Chiari malformation, and syringomyelia in seven patients born in breech position: a further proof of birth injury theory on the pathogenesis of "idiopathic hypopituitarism". We report seven cases of hypopituitarism all having a history of breech delivery, asphyxia at birth, and syringomyelia. A small pituitary gland was found on MRI or CT in six cases, invisible pituitary stalk on MRI in five cases, and type 1 Arnold-Chiari malformation in six cases. A constellation of these abnormalities are best explained by traction of brain and spinal cord of the subjects exerted during breech delivery and further support the primary role of birth trauma in the genesis of "idiopathic hypopituitarism".


Iatrogenic IgG2 deficiency in a leukaemic child. A case report. A girl with acute non-lymphoblastic leukaemia was treated with immunosuppressive chemotherapy. After cessation of therapy she had three consecutive episodes of infection due to Streptococcus pneumoniae from which she recovered and was shown to have developed a combined deficiency of both IgG2 and IgG4. The patient eventually relapsed and died 3 years after the initial diagnosis. The importance of measuring IgG subclasses in patients treated with immunosuppressive chemotherapy is discussed.


Spontaneous growth in Turner syndrome: evidence for a minor pubertal growth spurt. Spontaneous growth of 141 untreated girls with Turner syndrome was analysed. Of the patients 25% were born prematurely; their weight and height were normal when compared to prematurely born healthy infants. However, birth weight and height was significantly retarded in Turner patients born at term. A curve for height and growth velocity for the age range 0-16 years was constructed with a sensitive statistical method. By use of a mathematical model equations were created for calculating z-scores and the related percentiles for the height of individual patients at given age. Median height of 18 untreated patients at 18 years was 143.8 cm. Analysis of growth velocity revealed a minor but significant growth spurt at the age of 12.5 years. This growth spurt was also detectable in patients without signs of spontaneous puberty and occurred later in patients with 45,X0 karyotype. Bone age progression was linear up to the age of 7.5 years and decelerated thereafter.


IgG subclass deficiency in children with recurrent bronchitis. We studied the incidence of IgG subclass deficiency in children with recurrent bronchitis. Recurrent bronchitis was defined as three or more episodes a year during at least 2 consecutive years, of bronchopulmonary infection, productive cough with or without fever and/or diffuse râles by physical examination in the absence of asthma or atopy. Fifty three children were selected, of whom 30 (57%) were deficient in one of the IgG subclasses. None had an IgG1 deficiency. Nine (17%) were deficient in IgG2, 9 (17%) in IgG3 and 20 (38%) in IgG4. Isolated IgG subclass deficiencies were most frequently seen for IgG4 (14, 26%), less for IgG3 (6, 12%) and even less for IgG2 (4, 7%). Nine (17%) children were IgA deficient and 8 (15%) IgG deficient with a combined IgG subclass deficiency in 8 and 7 of them respectively. By subdivision into different age groups most patients were encountered in the youngest group. The mean content of IgG2, IgG3 and IgG4 in 3- to 4-year-old children with recurrent bronchitis was significantly lower than in the age matched controls. The mean value for IgG4 in the 5- to 6-year-olds was significantly lower than in the control group. This study demonstrates the correlation between recurrent bronchitis in childhood and IgG subclass deficiency. IgG subclass deficiency and recurrent bronchitis are both quite prominent phenomena in young children but rare in older children, suggesting a transient immaturity of the immune system as one of the possible pathogenetic factors. An IgA or an IgG deficiency is highly suggestive for the existence of a combined IgG subclass deficiency.


A family with one child with acrocallosal syndrome, one child with anencephaly-polydactyly, and parental consanguinity. A 2-year-old boy with typical features of the acrocallosal syndrome is presented. His parents are first degree cousins and their first pregnancy resulted in spontaneous abortion whereas the offspring of their second pregnancy was an anencephalic baby with bilateral post-axial polydactyly of the hands. The possibility of including anencephaly in the spectrum of the CNS malformations of the acrocallosal syndrome is suggested.


The relationship of fluid restriction during the 1st month of life to the occurrence and severity of bronchopulmonary dysplasia in low birth weight infants: a 1-year radiological follow up. One hundred consecutive low birth weight (LBW) infants (less than 1751 g) were randomized into a study group having a restricted fluid intake until 4 weeks of age and a control group following the fluid regimen conventionally used in the hospital. Chest X-ray films were examined on admission, at the ages of 3 days, 7 days, 2 weeks and 4 weeks and at bi-monthly visits to the out-patient clinic up to 1 year of age or until the chest examinations were normal. The severity of hyaline membrane disease (HMD) and typical radiological abnormalities of bronchopulmonary dysplasia (BPD) were assessed. Twelve patients succumbed, 1 in the study group and 11 in the control group. The study group seemed to experience less severe HMD than the controls. Of the former 54% and 32% of the latter were alive and had no radiographical signs of BPD at 4 weeks of age (P less than 0.05). The difference between the groups in the cumulative number of normal chest X-ray examinations during the follow up was even more significant. The percentage of normal X-ray films at 1 year of age was 92% in the study group and 72% in the control group. These results suggest that fluid restriction for the first 4 weeks of life can lower the incidence of radiological abnormalities typical of BPD obtained during the 1st year of life in LBW infants. Pulmonary oedema seems to be a significant aetiological factor causing HMD to develop into chronic lung disease.


Neonatal intestinal perforation due to congenital defects in the intestinal muscularis. Congenital defect of the muscular layer of the small intestine is a rare cause of spontaneous bowel perforation in premature infants. During the last 12 years we have observed four similar cases. We describe the most recent one, a premature infant who developed two abdominal events. On her 2nd day of life, spontaneous perforation of the distal ileum due to focal absence of the muscular layer occurred. Several weeks later she developed the typical clinical and histological picture of necrotizing enterocolitis. The clinical and histological characteristics of the two different conditions are compared, and the 24 cases reported in the literature are discussed. We conclude that focal absence of intestinal musculature may be not such a rare entity as is commonly believed.


Intra-uterine long bone growth in small-for-gestational-age infants. To better understand the intra-uterine bone modelling and remodelling process in small-for-gestational-age (SGA) newborn infants, long bone growth was studied using postmortem X-ray films in a group of such infants (n = 34). Bone length, diaphyseal diameter, medullary diameter, cortical thickness, cortical area, the Barnett-Nordin index, and the percentage of cortical area were determined in femur, tibia, and humerus. A separate group of appropriate-for-gestational-age (AGA) newborn infants (n = 146) was used as controls. Length and cortical bone mass in all three bones were significantly lower in SGA infants than in AGA infants. Decreased cortical bone mass in SGA infants was the result of decreased diaphyseal diameters and increased medullary diameters. Similar results were obtained when SGA infants were subclassified as preterm and term and compared with the control group of AGA infants. Bone lengths and diaphyseal diameters in SGA infants did not differ from those observed in a weight-matched control group of AGA infants although the latter were younger by 4 weeks' gestation. However, the cortical bone mass was lower than in the control group because of the relative greater medullary diameters in all three long bones in the SGA infants. Our present results indicate that reduced cortical bone mass in SGA infants is a mixed growth modelling and remodelling dependent process.


Asymptomatic urinary tract infection in childhood. In this study 2,591 apparently healthy children aged 1 day-16 years were examined to determine the existence of asymptomatic urinary tract infection. A prevalence of 4% in newborns, 5.2% in infants, 5.8% in preschool children, 4.5% in elementary school children, and of 4.8% in high-school children was found.


Identification of detomidine carboxylic acid as the major urinary metabolite of detomidine in the horse. Horse urine was investigated for metabolites by chromatography and mass spectrometry following the oral administration of the large animal analgesic sedative detomidine to two stallions and intravenous administration of [3H]-detomidine to a mare. Detomidine carboxylic acid and hydroxydetomidine glucuronic acid conjugate were identified in the urine after the oral doses. In addition, traces of free hydroxydetomidine were observed. About half of the radioactivity of [3H]-detomidine was excreted in the urine in 12 h after the i.v. dose (80 micrograms/kg). Most of the excretion occurred between 5 and 12 h in contrast to urine output which was highest 2-5 h after the dosing. The major radioactive metabolite in the urine was detomidine carboxylic acid. It comprised more than two thirds of the total metabolites in all the urine fractions collected. Its excretion profile was similar to that of total radioactivity. Hydroxydetomidine glucuronide was also excreted. It contributed 10-20% of the total metabolites in the urine. The free aglycone was only seen in the samples collected during the peak urine flow. A minor metabolite was tentatively characterized as the glucuronide of N-hydroxydetomidine.


Metabolic disposition of the non-steroidal anti-inflammatory agent isoxicam in man. The metabolic fate of isoxicam, a long half-life non-steroidal anti-inflammatory agent, in human subjects was investigated using isoxicam labelled with 14C in the N-methyl position. Three healthy male subjects were each administered a single oral 200 mg dose (90.7 microCi) with plasma and urine collected. Total plasma radioactivity peaked between 8 and 24 h postdose with mean 14C plasma radioactivity half-life of 36.1 h. Low levels of plasma radioactivity precluded plasma metabolic profiling. In urine, 37% of the administered radioactivity was recovered through 168 h. Metabolic profiling of urine confirmed the major oxidative excretion product as the hydroxymethylisoxazole metabolite. Identified and confirmed as minor urinary metabolites were radiolabelled open-ring sulfonamide and N-methylsaccharin. Non-labelled saccharin formed by oxidative loss of the 14C N-methyl group from N-methylsaccharin, was also observed. The role of this 'saccharin pathway' in the overall human disposition of isoxicam remains to be elucidated.


Clinical pharmacokinetics of the nifedipine/co-dergocrine combination in impaired liver and renal function. Following a single oral dose of 20 mg nifedipine combined with 2 mg co-dergocrine to 24 subjects, the pharmacokinetics of this drug were studied. 8 normotensive subjects had normal renal and hepatic function, 8 patients had chronic renal insufficiency (creatinine clearance less than 30 ml.min-1) and 8 patients had liver cirrhosis which was confirmed by liver biopsy. The area under the plasma level time curve (AUC infinity) of co-dergocrine increased from 0.59 +/- 0.41 ng.ml-1. (mean +/- SD) in the normals to 1.24 +/- 0.95 ng.ml-1.h in liver cirrhosis (P less than 0.05) and to 1.81 +/- 0.9 ng.ml-1.h in renal failure (P less than 0.05 compared with the control group). Corresponding values for the nifedipine AUC infinity were 564.5 +/- 268 ng.ml-1.h, 1547.5 +/- 1134 (P less than 0.05) and 929 +/- 533 ng.ml-1.h (P less than 0.05; gas chromatographic method). The incidence of adverse effects was lower in patients with renal failure than in subjects with normal renal and liver function as well as in those with liver cirrhosis.


Duration of postoperative immunosuppression assessed by repeated delayed type hypersensitivity skin tests. The duration of postoperative impairment in cell-mediated immunity was assessed by repeated skin testing with seven delayed type common antigens in 15 patients undergoing major elective abdominal surgery compared to a similar testing regimen in 10 healthy volunteers. All were skin tested four times, with 72-hour intervals, and in the surgical patients the first test was applied 2 days before surgery, followed by tests on postoperative days 1, 4 and 7. The tests were read after 48 h. Postoperatively, the skin test area decreased on day 3 (p less than 0.01) and recurred to preoperative levels on day 9. In contrast, the skin test area in the volunteers increased from test to test (p less than 0.001) during the study, confirming a previous finding of a vaccination effect. These results suggest that the postoperative immunosuppression is maintained for about 6-9 days.


Comparative bioavailability of Silipide, a new flavanolignan complex, in rats. The comparative pharmacokinetics of Silipide (IdB 1016, a silybin-phosphatidylcholine complex) and silybin were investigated by measuring unconjugated and total plasma silybin levels as well as total biliary and urinary silybin excretion in rats following administration of a single oral dose (200 mg/kg as silybin). Mean peak levels of unconjugated and total silybin after IdB 1016 were 8.17 and 74.23 micrograms/ml respectively. Mean AUC (0-6 h) values were 9.78 and 232.15 h.micrograms.ml-1 indicating that about 94% of the plasma silybin is present in a conjugated form. After administration of silybin, plasma levels of both unconjugated and total compound were under the analytical detection limit. Cumulative biliary (0-24 h) and urinary (0-72 h) excretion values after administration of IdB 1016 accounted for 3.73% and 3.26% of the administered dose, respectively. After silybin administration, the biliary and urinary excretion accounted for only 0.001% and 0.032% of the dose respectively. Our results indicate a superior bioavailability of silybin administered orally as IdB 1016. This was due mainly to an impressive increase in gastrointestinal absorption.


Effect of recurrent endotoxemia on hemodynamics, lung function and neutrophil activation in sheep. The aim of the study was to find out in which way lung permeability and polymorphonuclear leukocyte (PMNL) functions are modulated under recurrent endotoxin challenge, as it might occur in clinical septic patients. In a sheep model with chronic lung lymph fistula, performing bronchoalveolar lavage (BAL), we investigated the relationship between PMNL function and endothelial as well as epithelial damage in the lung in a sepsis syndrome, using a protocol of recurrent endotoxemia induced by 1 microgram/kg body weight Escherichia coli endotoxin treatment every 12 h over a 5-day period. Pulmonary response showed constantly increased pulmonary arterial pressure at mean values of 24-30 mm Hg. Also, lymph flow did not return to baseline, but remained on a level of 6-9 ml/30 min, after an increase to 12-15 ml/30 min following each endotoxin injection. In contrast, a lower increase in protein clearance was noted upon subsequent endotoxin administration. After initial values of 7-8 ml/30 min following the first endotoxin injection, almost baseline values were measured on the 5th day (3-4 ml/30 min). In systemic hemodynamics, we noted a decrease in cardiac output to 3.0 l/min after the first endotoxin injection, followed by a significant increase to 7 l/min under subsequent endotoxin administration. In PMNL function, we observed an attenuation of the acute response of the decrease in PMNL count, in vitro chemiluminescence response and plasma beta-N-acetylglucosaminidase level. The plasma urea concentration revealed a transient reduction in kidney function. In the epithelial lining fluid (ELF) of the alveoli, total cell count did not change significantly, but the fraction of PMNL increased from 2 to 20% during the 5 days. The ELF/plasma ratios of albumin and total protein did not change significantly. In conclusion, recurrent endotoxemia in a sheep model can produce a hyperdynamic state like in a sepsis syndrome which is further characterized by an initial leakage of the endothelial barrier, only minor affection of the epithelial barrier and by an exhaustion of PMNL function.


Suprahepatic vena cava anastomosis of the donor liver to the recipient retrohepatic vena cava in canine liver transplantation. A revised method of the upper caval anastomosis in canine orthotopic liver transplantation is described. It uses the recipient retrohepatic inferior vena cava below the suprahepatic veins for vascular suturing. Ten consecutive operations were performed to assess the feasibility of this method, with special reference to the outflow obstruction at the level of the suprahepatic inferior vena cava. Seven of 10 dogs survived more than 6 days. The cause of death was not related to the outflow obstruction in any instance. Regardless of the duration of survival, free hepatic vein pressures as well as portal vein pressures of all dogs remained in the physiological range even after the skin closures. Based on these observations, we conclude that this approach is safe and reproducible in experimental transplantation of the canine liver.


Influence of food on the oral bioavailability of moxonidine. Moxonidine is a new centrally acting anti-hypertensive with a very low adverse drug reaction profile. Among others, the aim of the study presented here was to determine the influence of food on the pharmacokinetics of moxonidine. Single oral moxonidine doses of 0.2 mg fasting and 0.2 mg non-fasting were administered in a randomized cross-over study. Eighteen subjects participated in the study, all of whom completed the study according to the protocol. Three sets of analytical plasma data could not be evaluated pharmacokinetically giving a total number of 15 evaluable cases. Renal excretion was evaluated for all 18 subjects. Food intake had no influence on the pharmacokinetics of moxonidine. The relative bioavailability of moxonidine administered under non-fasting conditions reached 94% of the bioavailability after fasted administration. Food intake resulted in a slight decrease of Cmax and a minimal increase of tmax as compared to the fasted treatment. The absorption half-life t1/2a showed a minor prolongation. These differences were not statistically significant in any of the parameters. For t1/2 lambda 2, CLtot and Ae(24h) no statistically significant differences were found between the fasting and non-fasting treatment. The amount of moxonidine excreted unchanged in urine accounted for about 46% of the dose administered after both treatments.


Metabolism of diazepam and related benzodiazepines by human liver microsomes. The metabolism of diazepam has been studied in vitro using microsomal preparations from five human livers. An HPLC method was developed for the assay of diazepam, its congeners and its metabolites. Various methods for the incorporation of diazepam into the incubation medium were explored. It was shown that the use of organic solvents or small quantities of hydrochloric acid enhanced the solubility of this substrate. However all of the organic solvents tested were associated with substantial (around 50%) inhibition of metabolism of diazepam by both major pathways (N-demethylation and C3-hydroxylation). The use of hydrochloric acid gave satisfactory solubilization of diazepam, but not of pinazepam, prazepam or halazepam. Detailed metabolic studies were conducted only for diazepam, using neither hydrochloric acid nor organic solvents in the incubation medium. Formation of N-desmethyl-diazepam increased approximately linearly with diazepam concentration to 200 microM, and did not show saturation. Formation of temazepam gave a curved profile over the same range of diazepam concentrations, suggestive of a sigmoidal relationship. Michaelis-Menten parameters could not be determined for either reaction, but intrinsic clearances for N-demethylation varied over a 6-fold range. Diazepam N-demethylation was apparently promoted by the inclusion of temazepam in the incubation medium, while C3-hydroxylation of diazepam was enhanced in the presence of N-desmethyldiazepam. Mephenytoin in the incubation mixture had no effect on diazepam metabolism by either pathway. The present studies have defined some of the methodological problems inherent in in vitro metabolic studies with benzodiazepines, and have shed further light on the metabolism of diazepam in vitro by human liver.


Tumor size, nuclear morphometry, mitotic indices as prognostic factors in axillary-lymph-node-positive breast cancer. The biopsy specimens from the primary tumors of 234 women with axillary-lymph-node-positive breast carcinomas (followed up for a mean of 10.9 years) were subjected to interactive morphometric analysis of nine nuclear factors. The proliferative activity of the tumors was estimated by determining two different mitotic indices. Morphometrically determined nuclear factors and mitotic indices showed a significant correlation to the histological grading (p less than 0.0001). Mitotic activity index (MAI; p = 0.018) and volume-corrected mitotic index (M/V index; p = 0.005) accurately predicted the tumor recurrence. Recurrence-free survival was related to the M/V index (p = 0.0003), MAI (p = 0.0024) and tumor size (p = 0.0144). Disease-related survival was determined by the tumor size (p less than 0.0001), M/V index (p = 0.0142) and MAI (p = 0.0492) in that order. On the other hand, the nuclear factors analyzed and the histological grading used had no predictive value (i.e. tumor recurrence, recurrence-free survival or tumor-related survival) in these women. The results indicate that mitotic indices can be successfully applied in place for subjective grading and nuclear morphometry in predicting the disease outcome in patients with axillary-lymph-node-positive breast carcinomas. The mitotic indices provide independent prognostic information in addition to tumor size. The major clinical implications of these results would be to accurately disclose among these women the high-risk patients (i.e. those with high mitotic indices), who might benefit from more agressive adjuvant therapies.


Changes in hepatic hemodynamics and oxygen consumption after partial hepatic congestion in dogs. The effects of 66% hepatic congestion (group 2, n = 6) on liver blood flow and hepatic oxygen metabolism were investigated in anesthetized dogs using an ultrasonic transit time flowmeter. The results were compared with those for control dogs (group 1, n = 6) and for 60% hepatectomized dogs (group 3, n = 6) wherein almost the same amount of hepatic parenchyma was removed as was congested in group 2. Portal blood flow (PVF) in group 2 and group 3 decreased similarly to 60 and 63% of the baseline values, respectively (p less than 0.05). Cardiac output (CO) in group 2 and group 3 also decreased significantly in proportion to the decrease in PVF. Among the dogs in group 2, hepatic arterial blood flow (HAF) was fairly well maintained at 86% of the baseline value, despite the decrease of cardiac output, whereas the HAF in group 3 decreased to 49% of the baseline value at 1 h after hepatectomy. The calculated hepatic arterial resistance (HAR) in group 3 increased significantly due to the 60% loss of the hepatic arterial vascular bed. The HAR in group 2, by contrast, became lower than that in group 1, suggesting a compensatory decrease of HAR for the obstructed portal flow to the congested area. These results were well consistent with our angiographic findings (n = 3) that the portal flow to the congested segments was completely obstructed and the congested segments received only an arterial blood supply. The centrilobular hepatocytes of the congested segments showed marked vacuolar degeneration and the total hepatic oxygen consumption in group 2 was reduced (p less than 0.05). However, the decrease in oxygen consumption in group 2 was not so severe as in group 3 (p less than 0.05). These data suggest that some parts of the preserved congested segments were still viable and had the capacity of aerobic metabolism even 4 h after the ligation of the drainage vein of those segments. In this study, the importance of the hepatic arterial flow to the congested segment has been demonstrated. When the congested hepatic segment is to be preserved intraoperatively, care must be taken to maintain the hepatic arterial blood flow during the perioperative period.


Absorption and disposition of 14C-labelled oxiracetam in rat, dog and man. The absorption and disposition of the nootropic drug oxiracetam (4-hydroxy-2-oxo-pyrrolidine-1-yl acetamide) were studied in rats and dogs (10 mg/kg i.v. and 10, 50 and 3000 mg/kg p.o.) and two healthy male volunteers (800 mg p.o.) using a [14C]-labelled preparation. Peroral absorption of [14C]-oxiracetam was incomplete in rats (28-42%), high in dogs (81-90%) and intermediate in man (about 56%). The rate of absorption was high in all species. Elimination was biphasic and the concentration of total radioactivity in blood and plasma declined rapidly with an initial elimination half-life of 1-3 h in all species. The specific systemic exposure to [14C]-oxiracetam was lowest in the rat, intermediate in the dog and highest in man. In all species the systemically available radioactivity was nearly exclusively excreted in urine in the form of unmetabolized oxiracetam. Whole-body autoradiography and quantitative determination of the radioactivity in various organs following i.v. and p.o. administration of [14C]-oxiracetam to rats demonstrated extensive distribution of the compound with high levels in kidney, liver, lung and skin, and very low levels in the brain. The radioactivity was rapidly eliminated from the body and minimal accumulation was observed upon repeated administration of 10 mg/kg for 8 days. Levels in the brain were still low, but higher than following a single dose, indicating slow diffusion across the blood-brain barrier. In pregnant rats treated with [14C]-oxiracetam radioactivity passed reversibly and to a limited extent through the placenta into fetal tissue.


Hyperthermo-radiosensitivity of esophageal cancer cells with high DNA ploidy in vitro. The in vitro sensitivity to hyperthermia and radiation of esophageal cancer cells obtained from 50 patients was assayed by using the in vitro succinate dehydrogenase inhibition test, and the findings were correlated with data on DNA analysis. The DNA distribution patterns were grouped into types I, II, III and IV, according to the frequency of aneuploid cell populations. Esophageal cancer cells of high ploidy (type IV) had a lower sensitivity to radiation, however, a greater sensitivity to hyperthermia as compared to cells of low ploidy (type II). The sensitivity to hyperthermia was determined as positive in 2 of 8 for type II, in 7 of 25 for type III, and in 9 of 17 for type IV. In contrast, the positive sensitivity rates to radiation in type II, III, and IV were 25.0, 8.0, and 5.9%, respectively. When cells were exposed to combination of radiation with hyperthermia, the positive sensitivity rates increased in all groups (50.0, 44.0 and 70.6% in type II, III, and IV, respectively). There was a significant correlation between mean DNA values and the SD activities following exposure to heat treatment or radiation. These data, describing that cancer cells of high ploidy might have greater sensitivity to hyperthermia compared to radiation, indicate the clinical benefits of hyperthermia in cases of esophageal carcinoma, especially for patients with high DNA ploidy.


Involuntary action of the external anal sphincter. Manometric and electromyographic studies. The changes in the rectal neck (anal canal) pressure and electromyography (EMG) of the external anal sphincter and levator ani muscle were studied in 20 dogs before and after internal anal sphincter excision by 2 weeks and monthly up to 10 months. The rectal neck pressure dropped to 41% of the preoperative level. It was then gradually elevated from the beginning of the 1st month following excision till it reached 88% of the level before excision at the 10th month. The external sphincter EMG basal activity after internal sphincterectomy by 2 weeks showed a decrease in the amplitude of the slow and high potential waves. From the 1st to the 7th month after excision, changes in the frequency and amplitude of the waves occurred, and by the 10th month, the external sphincter showed the characteristics of the EMG of both the internal and external anal sphincters. The basal activity of the levator ani muscle increased at the 10th month. As reported previously, the changes in the rectal neck pressure and in the EMG of the external anal sphincter after internal sphincterectomy seem to be due to the histological changes in the muscle and include striated and smooth muscle fiber hypertrophy and proliferation. These changes adapt the external sphincter to serve the function of not only voluntary but also involuntary continence.


Analysis of criteria for grading bladder cancer in urine cytological tumor diagnosis by means of an expert system. An inductive expert system was used for the analysis of criteria for grading bladder carcinoma in urine cytological tumor diagnosis. This analysis seems necessary in order to provide a better standardization of grading and to avoid tumor grades, which are rather inhomogeneous with respect to morphology and prognosis. The analysis of the database by the inductive system shows a considerable variation of the cytomorphology of different bladder carcinomas graded as G2 tumors, whereas G1 and G3 tumors are more homogeneous groups respectively. Especially nuclear morphological criteria are important features for the detection of highly differentiated carcinomas, whereas nucleolar features might be helpful to assess the proliferative nature of the carcinoma. The future goal of avoiding a grading system with prognostically inhomogeneous tumor grades seems possible when using an inductive expert system for consultation.


Ranitidine treatment and cortisol metabolism in man. Experimental evidence suggested that H2-receptor antagonists may inhibit not only hepatic but also adrenal cytochrome P-450 dependent monooxygenases. Therefore, the effects of ranitidine (150 mg b.i.d. over 14 days) on cortisol metabolism and antipyrine clearance have been investigated in nine healthy volunteers. Urinary excretion of 6 beta-hydroxycortisol (6 beta-OHC) and 17-hydroxycorticosteroids (17-OHCS) remained unaffected by ranitidine pretreatment, as did the calculated 6 beta-OHC/17-OHCS ratio. Only marginal effects were observed on antipyrine kinetics and metabolite formation. We conclude that neither adrenal production of corticosteroids nor their hepatic metabolism is affected by ranitidine administration.


Upper tract urothelial tumor after transurethral resection for bladder tumor. We present a retrospective review of 30 patients who developed upper urinary tract tumors (UUTT) after having been diagnosed and treated for a bladder neoplasm. The mean patient age was 63 years (range 54-72). An average of 4.8 (range 1-10) transurethral resections (TUR) had been performed for bladder cancer prior to the appearance of UUTT. The length of time elapsed between the initial bladder TUR and the subsequent diagnosis of UUTT was variable, with increased incidence in the first 24-72 months (63.3% of the cases). In 21 cases (70%) the bladder tumor was multiple and in 28 (93.3%) the tumor was recurrent. In those patients with unilateral vesicoureteral reflux, an increased incidence of UUTT was found in the refluxing renal unit. In patients with poorly differentiated (grade 3 and grade 2-3) bladder tumors, UUTT was of the similar grade of anaplasia in 75% of cases. These observations, together with those previously published in the literature, allow us to recommend the use of excretory urography (IVP) every 2 years during the first 6 years of follow-up in patients treated for recurrent and/or multiple bladder tumor. Thereafter, follow-up would depend upon the individual clinical situation.


Impotence due to corporeal veno-occlusive dysfunction: long-term follow-up of venous surgery. Corporeal veno-occlusive dysfunction is an important cause of organic impotence and is characterized by increased flow rates to create and to maintain erection during artificial erection produced by intracavernous saline infusion. Sixty-seven patients with this erectile insufficiency underwent penile ligature-resection of deep dorsal vein between 1982 and 1986, and were evaluated by nocturnal plethysmography, pharmacocavernometry as well as a questionnaire about their sexual life for long-term follow-up. The surgical procedure was controlled in the operating room by reduction of the erectile flow rates. Thirty-one patients were potent postoperatively and were able to have satisfactory intercourse. Results after resection were slightly better than after simple ligation of the deep dorsal vein. Four patients had penile glans insensibility resulting from the surgical dissection. There were 7 relapses several months after the procedure due to leakage through other deeper veins. Eight failures were due to additional psychogenic disorders or to neurologic disease not accurately diagnosed before the treatment because they all developed normal papaverine-induced erection after venous surgery while before it they only developed a slight tumescence. Reduction of the erectile flow rates within normal values was impossible during surgery in 3 patients. Eleven failures were due to concomitant arterial disease. Resection of the deep dorsal vein can restore penile erection in patients with cavernovenous leakage in about 50% of well-selected patients.


Penile revascularization in nonresponders to intracavernous injections using a modified microsurgical technique. At certain centers, microsurgical penile revascularization, using different surgical techniques, has gained importance throughout the past years. In general, only patients classified as intracavernous injection nonresponders are subjected to this kind of surgery. Since 1988, revascularization surgery has been performed at our clinic on 19 intracavernous injection nonresponders. The Hauri technique was carried out on the first 6 patients. The last 10 patients underwent modified anastomosis surgery. The inferior epigastric artery and the dorsal penile artery are anastomosed, one behind the other, end-to-side, to the dorsal penile vein. This results in a more simple procedure with assurance of flow. The Virag technique was performed on 3 patients. 18 patients achieved erections with or without the aid of intracavernous injections (at a mean follow-up of 13.4 months). 11 patients were capable of spontaneous erections, whereby it was particularly noted that 8 of the 10 patients undergoing the modified technique achieved spontaneous erections. The results demonstrate that intracavernous injection nonresponders benefit from revascularization surgery.


Can fine needle aspiration biopsy detect incidental prostatic carcinoma (T1) prior to TUR? The purpose of this study was to find out whether randomly taken fine needle aspiration biopsy (FNA) can detect incidental prostatic carcinoma prior to transurethral resection (TUR) and what are the effects of local tumor stage and grade on detection rate. Biopsies were taken from 344 patients, who came to hospitals for elective TUR without clinical evidence of prostatic carcinoma. Histologic examination of the TUR material showed prostatic carcinoma in 49 cases (14%). Sufficient material for cytologic examination was found in 343 cases. Of the 16 cases of T1a carcinoma in histologic examination, cytology found only 1, which was a G3 carcinoma. Of 33 T1b carcinoma in histologic examination, cytology found 6 and an additional 7 were suspect findings. Out of 6 G3 tumors in histologic examination, cytology showed 4. In our hands the proportion of false-negative cytologic findings in randomly taken FNA was so large that routine use of random FNA prior to TUR or as a screening procedure cannot be recommended, but positive FNA finding can be regarded as cancer.


Horseshoe kidney: the impact of percutaneous surgery. Six patients with a stone disease and/or ureteropelvic junction obstruction in a horseshoe kidney underwent percutaneous surgery. No major complications were observed and only 1 patient presented residual fragments in the lower calyx 3 months after treatment. The special features of the use of percutaneous nephrolithotomy and endopyelotomy for this anomaly are described.


Rationale of urodynamic assessment in adult enuresis. Between October 1988 and November 1990, 39 adult patients (average age 18.5 years) with enuresis underwent urodynamic evaluation. Filling cystometry was performed transurethrally using normal saline in the supine position. Abnormal cystometric findings were observed in 11 patients (28.2%), unstable bladder being the most common pathology (n = 7). Abnormal cystometric findings did not correlate with sex, age and diurnal symptoms of the patients. The only difference between pathologic and normal cystometry groups was the functional bladder capacity which is found to be reduced significantly in the former group. Cystometry does not contribute to the diagnosis of enuresis but helps to enlighten its physiopathology.


Effects of pentoxifylline in the hamster zona-free oocyte spermatozoa penetration assay and on spermatozoa transmembrane migration motility. The effects of pentoxifylline on capacitation, acrosome reaction and motility of spermatozoa in vitro were evaluated by the hamster zona-free oocyte penetration assay and the transmembrane migration method, respectively. The improvement in penetration capacity by the addition of pentoxifylline 40 min before coincubation had a trend toward statistical significance (0.05 less than less than 0.1) in normal fertile men and had statistical significance (p less than 0.05) in asthenospermic patients. There was no improvement by the addition of pentoxifylline 16 h before coincubation. Pentoxifylline significantly increased sperm motility in both normal (p less than 0.001) and asthenospermic patients (p less than 0.05) in the transmembrane migration method. This study showed that pentoxifylline could be used as a stimulant of in vitro sperm motility in male infertility.


Measurement of blood flow in the parameatal foreskin flap for urethroplasty in hypospadias repair. During the one-stage repair of hypospadias the blood flow of the parameatal-based foreskin flap was measured in 30 patients by using laser Doppler velocimetry. The mean blood flow in the penile foreskin was estimated to be 15 ml/min/100 g and it increased to 150-200% after the induction of anesthesia, and then decreased to 72% at the tip of the created parameatal foreskin flap. Topical application of papaverine hydrochloride restored the flow to the preincision level in half of the cases. The parameatal foreskin flap, both in the form of a manta-wing and in extended circumferential form, appears to have an enough microcirculation suitable for one-stage repair (OUPF II and OUPF IV, respectively) of hypospadias.


Erythropoietin in renal cell carcinoma: evaluation of its usefulness as a tumor marker. Erythropoietin levels in serum were analyzed in 165 patients with renal cell carcinoma. All samples were taken before therapy and stored at -80 degrees C. Erythropoietin, a glucoprotein produced by the renal cortex was quantified by an enzyme immunoassay. Fifty-five of 165 patients (33%) had elevated serum levels. In patients with metastatic disease (M+), elevated levels were found in 24 of 65 cases (38%). Patients with high-grade tumors had significantly more often increased erythropoietin than those with low-grade tumors. No correlation between erythrocytosis and elevation of erythropoietin in serum was found. There was a significant difference in survival between patients with normal and patients with elevated erythropoietin levels (p = 0.013). The study shows that erythropoietin is a tumor marker with a low sensitivity. However, it correlates with stage and grade and provides prognostic information.


Malignant priapism due to a huge renal carcinoma. Metastases to the penis from renal cell carcinoma producing priapism are very rare, as only 18 cases have been described since 1964. We present an additional case of priapism due to massive metastatic involvement of the corpora cavernosa in a patient with huge renal cell carcinoma, 28 cm in diameter, discussing the possible pathogenetic mechanism of such a rare condition.


Peritoneum-preserving radical nephrectomy in a patient on continuous ambulatory peritoneal dialysis. We describe a pararectus retroperitoneal approach for radical nephrectomy in a patient on chronic ambulatory peritoneal dialysis. This approach allows early ligation of the renal pedicle pre-operatively and immediate resumption of peritoneal dialysis post-operatively.


Malignant mesothelioma of the testicular tunica vaginalis. The histological, histochemical and immunohistochemical features of a malignant mesothelioma of the tunica vaginalis testis in a 69-year-old patient are described. The mesothelial derivation of the tumor was confirmed using a panel of selected antibodies. Malignant mesothelioma of the testicular tunica vaginalis is a rare tumor that must be considered so that appropriate treatment can be instituted.


Clinical significance of flow cytometric deoxyribonucleic acid measurements of deparaffinized specimens in bladder tumors. A retrospective study of flow cytometric measurements on paraffin-embedded tumor specimens from 188 patients with bladder tumor was conducted. Results were analyzed in combination with the morphological variation in bladder tumors. It was found that the deoxyribonucleic acid (DNA) ploidy pattern, the degree of infiltration and the multiplicity of bladder tumor were closely related with tumor recurrence, among which the DNA ploidy pattern was most significant. In aneuploid bladder tumors, the recurrence rate and the mean annual recurrence frequency were 76.7% and 1.46, respectively, and in the diploid bladder tumors, they were 18.7% and 0.33, respectively. Aneuploidy was the most indicative parameter of recurrence in bladder tumors. In addition, according to the DNA content and the ploidy level of the aneuploid cell lines, the aneuploid tumors in our group were divided into four types, namely tetraploid tumors, noneuploid tumors with an DNA index (DI) less than or equal to 1.5, hypotetraploid tumors and aneuploid tumors with several cell lines. The results showed that the recurrence rate of tetraploid tumors was relatively lower, and it became higher and higher in the following order: noneuploid tumors with DI less than or equal to 1.5, noneuploid tumors with DI greater than 1.5, and two-aneuploid tumors. This indicates that there are different biological behaviors in tumors with different ploidy patterns. Finally, the correlation between the DNA ploidy pattern and tumor metastases was also discussed.


Extracorporeal shock wave lithotripsy with ultrasound-guided lithostar plus. Since 1989, the Siemens lithostar plus, an upgrade of the lithostar with the ultrasonically guided overhead module, has been available for clinical use. This unit may be used for the treatment of either biliary or urinary calculi. We report on 75 patients with urinary calculi treated between March 1989 and June 1990 with the lithostar overhead module. Stone localization showed a rate of: caliceal stones 33.5%, pelvic stones 44%, upper ureteral stones 9.3% and lower ureter stones 13%. The overall disintegration rate was 86%, with a stone-free rate after 3 months of 78%.


Topical chemoprophylaxis of superficial bladder cancer with mitomycin C and adjuvant hyaluronidase. The effect of intravesical instillation of 200,000 IU hyaluronidase in addition to mitomycin C as chemoprophylaxis of superficial bladder cancer was evaluated. Based on our result of a previous study that this approach results in a significantly reduced recurrence rate (7.1 versus 32.1%), 43 patients undergoing transurethral resection of Ta-T1 tumors were retrospectively analyzed after a mean observation period of 48.5 months. During the 2 years of prophylactic therapy, tumor recurrence was seen in 6 patients (13.9%). Of the 37 patients who remained recurrence-free under treatment, 5 (13.5%) had recurrent tumors later during a mean observation period of 24.5 (18-42) months after treatment. These values are significantly lower than those obtained previously from a group of 63 patients treated with mitomycin alone, with recurrence rates of 33.3% during chemoprophylaxis and 26.2% thereafter (mean observation period 50.4 months). It appears that additive hyaluronidase enhances the local effect of mitomycin C in the intravesical chemoprophylaxis of bladder cancer, presumably by improving diffusion into the bladder mucosa and catalyzing the breakdown of a hyaluronic acid contained in a protective halo around malignant urothelial cells.


Treatment of advanced bladder cancer by submucosal injection of methotrexate in the anorectal area. In 1988, Shafik et al. presented their first attempt to apply cytostatics in the treatment of pelvic malignancies. Experimentally and clinically, they studied the anal route for administration of methotrexate, and their first results showed that higher concentration of drugs can be achieved in the bladder wall compared to that in serum. Since December 1988, we have been engaged in experimental and clinical investigations of the possibilities this method might reveal. Experiments with animals have proven that submucosal injection of the cytostatic agent does not induce local changes. The clinical trial on 20 patients with tumors of stage T2-T4 has not shown side effects, either local or systemic. The treatment was well tolerated by the patients. Anorexia has been reported in 3, elevated temperature in 2 and leukopenia in 4 of 20 patients. Complete response has been documented in 3 (15%), partial response in 3 (15%), stabilization in 5 (25%) and progression of the disease in 9 (45%) of the patients. Mean duration of remissions was 10.3 months (range 9-12). The distribution of submucously injected contrast medium into the anorectal area has been studied by computerized tomography in 4 of the patients. Although our results do not support the original estimates of the inventors of the method, its ease of application, the possibility of out-patient administration and negligible side effects are sufficient arguments for performing further experimental and clinical studies.


Frequency of urogenital tuberculosis in an unselected autopsy series from 1928 to 1949 and 1976 to 1989. The evaluation of a total of 12,211 autopsy protocols prior to the introduction of chemotherapy (1928-1949) and in the recent years (1976-1989) had the following results: the frequency of renal tuberculosis has dropped from 2.0 to 0.6% of all autopsies in men and from 1.7 to 0.9% in women. While the frequency of miliary tuberculosis infections in general has decreased to about one fourth (from 2.7 to 0.6% of all autopsies), renal involvement in patients with miliary tuberculosis has rather increased (from 35.4 to 52.2 for men and from 37.5 to 68.0% for women). The frequency of miliary tuberculosis of the kidneys was two times higher in girls than in boys (3.8% of girls and 1.8% of boys). Tuberculosis of the renal pelvis, ureter or urinary bladder secondary to a renal tuberculous infection has decreased from 65 to 4.3% of all tuberculous infections with renal involvement in males and from 25 to 0% in females. In the second period, the frequency of genital tuberculosis has in all dropped to about one tenth.


The Wardill test: does it predict future voiding flow? A prospective study was undertaken to assess the predictive value of measuring the peak flow rate at completion of a transurethral prostatectomy with suprapubic pressure (the Wardill test). Twenty-six patients had their expressed flow rate measured on applying a standardised suprapubic pressure and compared to their peak flow rate measured at least 6 weeks postoperatively. No significant correlation was found to exist between the two variables.


Inhibitors of stone formation in hypercalciuric children with and without stone disease. The extretion of two inhibitors of urinary stone formation (citrate, magnesium) was evaluated in 71 hypercalciuric children and 50 controls. Patients were classified into two groups: 42 nonstone former (NSF), 22 boys and 20 girls, 3-14 years old, and 29 stone formers (SF), 18 boys and 11 girls, 2.5-18 years old. Our study was unable to show significant differences in magnesium and citrate urinary outputs between controls and patient groups. The Mg/Ca ratio was found significantly lower in hypercalcuric children than in controls (p less than 0.001), but not between NSF and SF patients. Our data demonstrated that both NSF and SF groups had a significantly lower citrate/Ca ratio than controls (p less than 0.001), also it was lower in SF than in NSF (p less than 0.05). We found no significant difference in citrate excretion between boys and girls neither in patients nor in controls.


Histopathological alterations of deep dorsal penile vein in venogenic impotence. The mechanism and hemodynamics of penile veins in erection have long been a matter of controversy and hypotheses. With the intention to develop a new concept of venous ultrastructure in penile venous insufficiency, we studied the histopathology of 13 dorsal penile veins obtained from important patients during venous ligation because of proven venous leakage. Besides, 1 normal deep dorsal vein taken from a volunteer who underwent another type of penile operation was also examined. All sections were stained with hematoxylineosin, Gomori trichrome, periodic acid-Schiff, Gomori reticulum and Verhoeff elastic stains. All 13 veins showed some significant histological changes namely nodular hypertrophy in 7, nodular atrophy in 2, diffuse atrophy in 2, diffuse hypertrophy in 1 and minimal changes in 1. In general, the dominating histological appearance was hypertrophy of the muscular pattern and enhancement of collagenization. None of these alterations was prevalent in the normal control. The cause and the result relationship between venous leakage and the mentioned histological changes still remains a matter of debate.


Effect of external beam radiotherapy on prostatic carcinoma DNA content as measured by static image cytometry. Static image cytometry was used to study the effect of radiotherapy on the DNA content of prostatic carcinoma. Feulgen-stained specimens from 50 patients before and after radiotherapy were examined. The DNA index increased significantly after treatment (p less than 0.03), and it occurred equally in those with low (less than 1.5) and high (greater than or equal to 1.5) indices. The level of the initial DNA index was significantly related to the rate of symptomatic progression (p less than 0.05). The initial change in and final DNA indices were not related to survival however. There was a significant increase in Gleason score (p = 0.001) and histological grade (p = 0.01) after irradiation and, when taken individually, the degree of change was related to survival in both, p = 0.05 and 0.004, respectively. As Gleason score, Mayo grade and clinical stage were not independent variables; none of them was related to survival when corrected for the other factors. Thus, DNA content or ploidy, as well as Mayo grade and Gleason score, increases in recurrent tumor after radiotherapy. This may be of importance when considering what treatment to use and how intensively the patients should be followed up.


The role of serum laminin P1 in the diagnosis of transitional cell carcinoma of the bladder. In order to explore the possible role of serum laminin P1 in the diagnosis of transitional cell carcinoma (Tcc) of the bladder, the sera of 50 healthy control subjects and of 35 bladder-cancerous patients were measured by radioimmunoassay. In 27 patients (77%), the serum level was elevated above the upper limit of normal range. Statistically significant elevation could be determined in all stages and grades of Tcc of the bladder compared to those of normal subjects (mean +/- SD: 1.18 +/- 0.16 kU/l). Deterioration of the clinical stages or pathologic grades of the tumor was associated with a progressive increase in the mean values of serum laminin P1. A longer study with serial determination is planned to assess the prognostic significance of serum laminin in bladder cancer patients.


A system for focal intracavitary irradiation of bladder cancer with remote iridium-192 afterloading. An afterloading system for remote-control focal intracavitary irradiation of bladder cancer is presented. With 192Ir as radiation source, we demonstrated the practicability of the method in 23 chinchilla rabbits. A system for focal intracavitary irradiation of bladder cancer in humans was developed. A guiding tube with an inflatable balloon led to a reproducible vesical diameter. The intravesical part of the tube was shielded with lead and had a slit for focal radiation exposure. Three variant guiding tubes with different positions of the slit were designed to allow irradiation of any part of the bladder. The application is planned in patients with locally advanced tumors who refuse radical cystectomy or who are medically inoperable.


Studies on the retention of the mucous-membrane-adhesive anticancer agent hydroxypropylcellulose doxorubicin. Incorporation of hydroxypropylcellulose (HPC-)doxorubicin, which we developed as a mucous-membrane-adhesive drug preparation, was instilled into the urinary bladder in 10 clinical cases. Tumor of the urinary bladder was a single tumor in all 10 cases, and preclinical histology showed transitional cell carcinoma, grade 1 or 2, and a lower stage than T1. HPC-doxorubicin, 20 mg/20 ml, was administered in 5 cases, and the other 5 cases received the conventional aqueous doxorubicin, 20 mg/20 ml by way of a catheter and the urethra. Cold punch biopsy was performed after 3 days of instillation, and the incorporation of doxorubicin into both tumorous and normal tissue was measured by high-pressure liquid chromatography. After 3 days, it was found that in the HPC-doxorubicin-administered group, doxorubicin was detected in both tumorous and normal tissue, but it was not detected in either tissue after aqueous doxorubicin administration. In 5 cases of the HPC-doxorubicin group, doxorubicin levels in the tumorous and normal tissue were examined, and it was found that significantly more doxorubicin was detected in the tumorous tissues. Thus, it may be said that our HPC-doxorubicin remained longer within the urinary bladder than the conventional aqueous doxorubicin preparation. Instilled HPC-doxorubicin is more highly concentrated in the tumorous tissue than in the normal bladder tissue, and thus, HPC-compounded anticancer drugs may be therapeutically more useful.


The human renal 25-hydroxyvitamin D3-1 alpha-hydroxylase: properties studied by isotope-dilution mass spectrometry. The renal 25-hydroxyvitamin D3-1 alpha-hydroxylase activity has been measured in normal human kidney cortex, using a highly specific assay based on isotope-dilution mass spectrometry. The cortex was obtained from kidneys removed due to renal tumours. The subcellular distribution of 25-hydroxyvitamin D3-1 alpha-hydroxylase activity was studied. Enzyme activity was only observed in the mitochondrial fraction. Mitochondria from non-tumourous kidney cortex had a Vmax of 0.17 +/- 0.02 pmol min-1 mg-1 protein and the apparent Km was in the range of 14 mumol l-1. There was a tendency to a higher 25-hydroxyvitamin D3-1 alpha-hydroxylase activity in preparations from male kidney (0.21 +/- 0.03 pmol min-1 mg-1 protein) than female (0.12 +/- 0.02, P less than 0.05). A significant inverse correlation between serum phosphate and 25-hydroxyvitamin D3-1 alpha-hydroxylase activity was found. No correlation was observed between enzyme activity and serum levels of 1,25-dihydroxyvitamin D (total and free index), PTH, total calcium or ionized calcium. The results indicate that there is a sex difference in human 25-hydroxyvitamin D3-1 alpha-hydroxylase activity similar to the one observed in laboratory animals. Furthermore, the data support the hypothesis that serum phosphate is a major regulator of 1,25-dihydroxyvitamin D3 production in man.


Uptake of chylomicron remnant retinyl esters in human leukocytes in vivo. Retinoids have been successfully used in the treatment of some forms of leukaemia, suggesting that such cells have an efficient uptake mechanism for circulating retinoids. Therefore, we have studied the uptake of lipoprotein-associated retinyl esters in human leukocytes in vivo. After an oral load of 100 mumol retinyl palmitate (30,000 retinol equivalents) per square meter given to healthy adults, the concentration of retinoids in circulating leukocytes was determined. A peak was measured after 5 h, which coincided with a peak of retinyl esters in plasma. To test whether low-density lipoprotein receptors are necessary for the postprandial uptake of retinoids, we studied retinoid uptake in leukocytes from two patients homozygous for familial hypercholesterolaemia. After an oral load of retinoids we found that leukocytes from these patients took up at least as much retinoid as leukocytes in normal individuals, suggesting that uptake of chylomicron remnant retinyl esters may proceed independent of the low-density lipoprotein receptor. The expression of mRNA for the low density lipoprotein receptor-related protein, which is a putative chylomicron remnant receptor, was similar in leukocytes from a patient homozygous for familial hypercholesterolaemia and normal individuals. Six hours after vitamin A administration, recovery of unesterified retinol was 71% in normal leukocytes, however, only 9% unesterified retinol was recovered in leukocytes from the two patients with familial hypercholesterolaemia. Thus, the apparent rate of retinyl ester hydrolysis was markedly reduced in leukocytes from these patients, indicating different intracellular traffic of chylomicron remnants in normal individuals and patients homozygous for familial hypercholesterolaemia.


Incomplete suppression of hepatic glucose production in non-insulin dependent diabetes mellitus measured with [6,6-2H2]glucose enriched glucose infusion during hyperinsulinaemic euglycaemic clamps. We have minimized methodological errors in the isotope dilution technique by using stable isotope, [6,6-2H2]glucose, thus avoiding the problem of contamination of tritiated glucose tracers and, by maintaining a constant plasma tracer enrichment have reduced error due to mixing transients. Using these modifications we have calculated hepatic glucose production in 20 patients with non-insulin-dependent diabetes mellitus during low (1 mU kg-1 min-1) and high (8 mU kg-1 min-1) dose insulin infusions. Mean fasting hepatic glucose production was 14.2 +/- 0.8 mumol kg-1 min-1. This suppressed by only 68% to 4.6 +/- 0.8 mumol kg-1 min-1 during the low-dose insulin infusion (plasma insulin 0.85 +/- 0.05 nmol l-1) and did not suppress further during the high-dose insulin infusion (plasma insulin 14.55 +/- 0.83 nmol l-1). Hepatic glucose production was significantly higher than zero throughout the study. Thus, we have found that minimization of known errors in the isotope dilution technique results in physiologically plausible and significantly positive values for hepatic glucose production indicating that the liver is resistant to insulin in patients with non-insulin-dependent diabetes mellitus.


Identification in human urine of a natural growth inhibitor for cells derived from solid paediatric tumours. Partially purified urine of healthy human subjects contains several fractions able to inhibit the proliferation of cultured human neuroblastoma cells. One of the most active fractions was further analysed by gas chromatography-mass spectrometry and shown to contain genistein, a substance formed in the human body from precursors obtained by diet. Synthetic genistein was able to inhibit the proliferation of human neuroblastoma cells with a half-maximal effect at 5-10 mumol l-1 concentrations. Genistein displayed similar potencies in inhibiting the proliferation of cells derived from various other solid pediatric tumours. Our results suggest that genistein is a natural antineoplastic agent present in diet and that it could be useful for the therapy of paediatric tumours.


Human recombinant C5a enhances lipopolysaccharide-induced synthesis of interleukin-6 by human monocytes. The effect of human recombinant C5a (hrC5a) on the synthesis of interleukin-6 (IL-6) was studied in human monocytes. Monocytes incubated in the absence of hrC5a and of bacterial lipopolysaccharide (LPS) produced only low amounts (less than 100 U/2 x 10(6) cells/16 h) of IL-6 activity. LPS in concentrations from 10 pg ml-1 to 10 ng ml-1 greatly stimulated the synthesis of IL-6 to about 50.000 U/10(6) cells/16 h. When hrC5a was added to the monocyte media maximal IL-6 synthesis was reached at lower LPS concentrations, i.e. at 0.1 ng ml-1 LPS in the presence of 100 ng ml-1 hrC5a. Maximal IL-6 production was not significantly enhanced by hrC5a. Metabolic labelling with [35S]-methionine followed by immunoprecipitation of IL-6 showed that the increased IL-6 activity in the medium of hrC5a treated monocytes was due to a stimulation of the de novo synthesis of IL-6. Increased amounts of IL-6 mRNA were found in monocytes treated with LPS and hrC5a compared with monocytes stimulated only with LPS. HrC5a prolonged the elevation of IL-6 mRNA levels after stimulation of monocytes with LPS. HrC5a thus enhanced the LPS-induced synthesis of IL-6 by human monocytes.


Regional effects and clearance of endothelin-1 across pulmonary and splanchnic circulation. To determine the impact of i.v. endothelin-1 on systemic, pulmonary and splanchnic circulation, as well as the peptide's regional clearance, hepatic venous and right heart catheterization was performed in healthy volunteers. During the peptide's continuous i.v. administration (0.4 pmol x kg-1 x min-1, 60 min) its plasma concentration rose from 2.1 +/- 0.5 to 9.5 +/- 5.3 pmol/l (pulmonary artery), from 2.1 +/- 0.9 to 5.0 +/- 1.6 pmol/l (femoral artery), and from 1.5 +/- 0.6 to 2.9 +/- 1.2 pmol/l (hepatic vein). This was accompanied by an increase in mean systolic arterial pressure from 127 +/- 14 to 131 +/- 12 mmHg (P less than 0.05). Concomitantly, cardiac output and heart rate decreased from 7.0 +/- 1.1 to 5.8 +/- 1.0 l/min and from 63 +/- 6 to 56 +/- 5 beats/min, respectively, while total vascular resistance increased from 964 +/- 273 to 1204 +/- 338 dyn x cm x s-5 (P less than 0.01). No major changes in pulmonary circulation were observed, while splanchnic vascular resistance increased from 4472 +/- 1056 to 5361 +/- 1420 dyn x cm x s-5 (P less than 0.01) and estimated hepatic blood flow decreased from 1403 +/- 218 to 1218 +/- 219 ml min-1 (P less than 0.01). During endothelin-1 infusion the pulmonary vascular bed accounted for approximately 53% of the peptide's overall disposal.(ABSTRACT TRUNCATED AT 250 WORDS)


Glucagon-like peptide-1 cells in the gastrointestinal tract and pancreas of rat, pig and man. A highly specific monoclonal antibody directed against the C-terminal part of glucagon-like peptide-1 (GLP-1) was raised to immunohistochemically evaluate the distribution of GLP-1 containing cells in the entire gastrointestinal tract including pancreas of rat, pig and man. In the pancreas GLP-1-immunoreactive cells were found variously shaped and predominantly located in the periphery of the islets. Ultrastructurally, GLP-1 was co-localized with glucagon in the alpha-granula of A-cells and was mainly restricted to the electrondense core. In the intestine open type cells reaching the lumen via a slender apical process were stained with the GLP-1 antibody. They occurred in all parts of the crypts but predominantly in the basal portion. The density of GLP-1 immunoreactive cells varied between species in a characteristic order: rat greater than pig greater than man. In pig and human gut a large number of cells occurred in the distal jejunum and ileum. A continuous increase of cell densities was found from the proximal to the distal colon resulting in highest numbers in the rectum. In rats the highest cell density occurred in the ileum. Again, a continuous increase of GLP-1-positive cell numbers was evident from the proximal to the distal portion of small and large bowel. GLP-1 was partly co-localized with PYY. The GLP-1 positive cells appeared electronmicroscopically as L-cells with the typical large granula. This morphological data indicates that GLP-1-releasing cells in the small intestine are appropriately positioned in the distal part to sense and respond to the presence of nutrients that have escaped the absorptive surface of the upper small intestine.


Effects of interferon alpha on insulin binding and glucose transport in human adipocytes. We have previously demonstrated that interferon administration impairs glucose tolerance and causes insulin resistance in healthy man. Whether this is a direct effect of interferon is not known. The present study was undertaken to examine directly the effect of interferon alpha on insulin binding and action on glucose transport in isolated human adipocytes. Different concentrations of interferon alpha (range 10(-3)-10(5) IU ml-1) and different incubation times (0-5-24 h) with interferon were employed. Acute and 5-h and 24-h exposure of human adipocytes to 10(-2)-10 IU ml-1 of interferon increased the high affinity binding of 125I-insulin (P less than 0.05). In contrast, human interferon alpha had no effect on insulin binding in rat adipocytes. In short-term studies interferon had no effect on 14C-glucose transport clearance. 24-h preincubation of human adipocytes with 10(-2), 10, 10(4) IU ml-1 interferon increased maximally-insulin stimulated 14C-glucose transport clearance (P less than 0.05) and glucose transport responsiveness to insulin was enhanced by 24% (P less than 0.05) in cells exposed to 10(-2) IU ml-1 interferon. After 5 and 24-h preincubations with interferon we observed modest changes in glucose transport sensitivity to moderate concentrations of insulin (50-100 pM) with upregulation in the presence of 10(-2)-10 IU ml-1 interferon and downregulation in the presence of 10(4)-10(5) IUm ml-1 interferon (P less than 0.05). The insulin sensitivity index (ED50) did not change.(ABSTRACT TRUNCATED AT 250 WORDS)


Neurogenic responses of urethra isolated from the dog. Electrical transmural stimulation evoked contraction and relaxation in isolated urethral circular muscle of the dog. The responses were abolished by tetrodotoxin, indicating their neurogenic origin. The contractile force in the middle urethra was greater than that in the proximal and distal urethra. The contractions were not affected by atropine and propranolol, but were completely inhibited by phenoxybenzamine, prazosin and guanethidine. In preparations contracted with prostaglandin F2 alpha, electrical stimulation induced frequency-dependent relaxation in all urethral portions. Atropine, phenoxybenzamine, prazosin and guanethidine had no effect on the relaxation, while propranolol slightly attenuated the relaxation induced at the highest frequency used (5 Hz). The non-adrenergic, non-cholinergic relaxation was also not affected by ketanserin, methysergide, diphenhydramine, alpha,beta-methylene ATP or capsaicin. Exogenously applied phenylephrine and clonidine both produced contractions but the maximal response to clonidine was much smaller than that to phenylephrine. Acetylcholine produced no or feeble contractions. In the preparations contracted with prostaglandin F2 alpha, isoproterenol and vasoactive intestinal polypeptide (VIP) produced relaxation. These results suggest that the circular muscle of dog urethra is reciprocally innervated by sympathetic adrenergic and non-adrenergic, non-cholinergic nerves, and that the neurogenic responses are markedly affected by muscle tension and the portion of the urethra examined.


Characterization of the peripheral action of neuropeptide K on the rat cardiovascular system. The effects of neuropeptide K (NPK) were measured on mean arterial pressure (MAP) and heart rate (HR) after i.v. injection in urethane-anesthetized rats. NPK (6.5 and 32.5 nmol/kg) produced sustained decreases in MAP and elicited increases in HR. Whereas the NPK-induced tachycardia lasted more than 30 min at 32.5 nmol/kg, a latent and long-lasting bradycardia appeared from 20 min after injection of 6.5 nmol/kg. The initial tachycardia was converted to bradycardia by metoprolol but remained unaffected by hexamethonium, atropine and naloxone. These four treatments, however, prevented the bradycardiac response to NPK at 30 min. Whereas phentolamine, idazoxan, bilateral adrenalectomy and chemical sympathectomy with 6-hydroxydopamine (6-OHDA) preserved the initial tachycardia induced by NPK, they converted the decrease in HR to a tachycardiac response at 30 min. The vasodepressor response to NPK was significantly enhanced by bilateral adrenalectomy, chemical sympathectomy and metoprolol but remained unaffected by all other treatments. Neither the MAP nor the HR responses to NPK were affected by indomethacin. These results suggest that NPK can accelerate HR through non-reflex activation of the sympathoadrenal system. The secondary bradycardia induced by NPK may be due to a vagal reflex while the vasodepressor response to NPK is probably attributable to a direct action mediated by specific receptors on arterial blood vessels. Thus, NPK is considered as the most potent biologically active tachykinin so far described on the rat cardiovascular system.


The protein kinase C activators, phorbol 12-myristate,13-acetate and phorbol 12,13-dibutyrate, are convulsant in the pico-nanomolar range in mice. Administration of phorbol 12-myristate,13-acetate (PMA, 10 fmol-10 nmol) or phorbol 12,13-dibutyrate (PDB, 0.2-495 nmol) (i.c.v.) to mice induced: hindlimb scratching, tremor, myoclonic jerks, hyperlocomotion, clonic seizure, followed by death or recovery. CD50 values for clonic seizures for PMA and PDB were 1.0 pmol and 1.2 nmol. 4-alpha-Phorbol (68-686 nmol) was inactive. The effects of PDB (24-247 nmol) were reduced by pretreatment with staurosporine (30 nmol, i.c.v.). Protein kinase C activators are potent convulsants in vivo.


The effect of hydrochlorothiazide on the enhanced coughing associated with treatment with enalapril. The effect of hydrochlorothiazide, a diuretic which is used in the treatment not only of edema but also of hypertension, on coughs associated with treatment with enalapril was studied in guinea pigs. Chronic treatment with enalapril markedly and dose dependently enhanced the number of capsaicin-induced coughs. However, chronic treatment with hydrochlorothiazide significantly reduced the number of coughs associated with enalapril treatment, also in a dose-dependent manner. These results suggest that diuretics might be used to reduce the coughing associated with treatment with inhibitors of angiotensin-converting enzyme in patients with hypertension.


External ATP antagonizes the effect of potassium channel openers in guinea-pig ventricular papillary muscle. Right ventricular papillary muscles of the guinea-pig heart were electrically stimulated. Cromakalim 10-100 microM and Ro 31-6930 3 microM depressed the contractile force and shortened the duration of action potentials. Glibenclamide 0.3-3 microM, ATP 100 microM and alpha, beta-methylene ATP (alpha, beta-meATP) 30 microM antagonized these effects. Suramin 300 microM failed to reverse the alpha, beta-meATP-evoked antagonism of the action of cromakalim. It is concluded that both intra- and extracellular ATP may interfere with potassium channel openers and that extracellular ATP does not act via the known P2-purinoceptor subtypes.


CCK-8-stimulated insulin secretion in vivo is mediated by CCKA receptors. The effects of the cholecystokinin A (CCKA) receptor antagonist, L-364,718, and the CCKB receptor antagonist, L-365,260, on CCK-8-stimulated insulin secretion were studied in vivo in the mouse. It was found that CCK-8-stimulated insulin secretion was suppressed by L-364,718 at a low dose level (0.078 mumol/kg). In contrast, L-365,260 caused a partial inhibition of CCK-8-stimulated insulin release only at the high dose level (24 mumol/kg). It is concluded that the CCK-8-stimulated insulin release in vivo is mediated by CCK receptors of the CCKA subtype.


Glibenclamide does not block arterial relaxation caused by vasoactive intestinal polypeptide. Vasoactive intestinal polypeptide (VIP) caused a concentration-dependent relaxation in rabbit mesenteric artery with intact endothelium. The relaxant response to VIP was inhibited by the removal of the endothelium or by pretreatment with methylene blue, but not by pretreatment with glibenclamide. A small but significant relaxant response to VIP in the endothelium-denuded artery was also unaffected by glibenclamide. These findings indicate that ATP-sensitive K+ channels are not involved in the endothelium-dependent or endothelium-independent arterial relaxation elicited by VIP.


Probenecid pretreatment enhances anticonvulsant action of NBQX in mice. NBQX (2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline) was studied following intravenous administration to mice. Maximal electroshock seizures were suppressed by low doses of NBQX (ED50 = 13 mg/kg) but effects had dissipated by 10 min after a dose of 25 mg/kg. Coadministration of the transport inhibitor probenecid (p-(dipropylsulphamoyl)-benzoic acid) enhanced and prolonged the anticonvulsant action of NBQX and also enhanced and prolonged ataxia. NBQX may be rapidly eliminated by a process sensitive to probenecid.


Effect of haloperidol and clozapine on (+)SKF 10,047-induced dopamine release: role of 5-HT3 receptors. (+)SKF 10,047 preferentially increased dopamine release in the nucleus accumbens compared to the striatum. Dopamine output was evaluated in the same freely moving rats by trans-cerebral dialysis. Clozapine and DAU 6215, a 5HT3 antagonist, which itself did not modify dopamine release in both areas, selectively antagonized (+)SKF 10,047-induced dopamine release in the nucleus accumbens. Haloperidol by itself increased dopamine release in both areas and these effects were additive with those induced by (+)SKF 10,047.


Dopamine release and dopaminergic inhibition of acetylcholine release in rat striatal slices after nigro-striatal hemitransection and parenteral ganglioside administration. Hemitransection of the nigro-striatal bundle in adult rats reduced [3H]dopamine ([3H]DA) uptake into striatal slices from the lesioned side to about 20% of that in the contralateral side 5 days after surgery. Spontaneous recovery of [3H]DA uptake was observed at days 8 and 15 post-lesion (42 and 67% of the unoperated side, respectively). After a short treatment (3 days) with the GM1 ganglioside inner ester (AGF2, 30 mg/kg i.p., daily, starting on day 2 after surgery) [3H]DA uptake amounted to 52% of that in the unoperated side. The electrically evoked fractional overflow of [3H]DA was increased by 500% in slices prepared from the lesioned side 5 days after injury, largely due to the reduced re-uptake by the DA axon terminals. The increase on day 5 was only about 350% in AGF2-treated animals. The DA D2 receptor antagonist, (-)-sulpiride, potentiated the stimulus-evoked overflow of [14C]acetylcholine in slices from the unoperated side prelabelled with [14C]choline. The effect of (-)-sulpiride was much reduced (by about 80%) in the lesioned striata at days 5 and 8 after surgery. Partial recovery was seen at day 15. The lesion did not modify the (-)-sulpiride effect in animals treated with AGF2 from the 2nd to the 5th day post-lesion. Thus early ganglioside administration slows the loss of endogenous dopaminergic control of acetylcholine release caused by partial hemitransection of the nigro-striatal bundle.


Mechanisms involved in the hyperglycemic effect of the 5-HT1C/5-HT2 receptor agonist, DOI. Previous experiments have indicated that 5-HT2 receptors and catecholaminergic systems mediate the rise in plasma glucose levels elicited by acute administration of the 5-HT1c/5-HT2 receptor agonist, 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane (DOI). On this basis, we investigated the location of these serotonin receptors and the nature of this catecholaminergic involvement. Administration of DOI (0.4 mg/kg i.v.) to conscious rats (bearing jugular catheters) elicited a rapid rise in plasma glucose which was associated with a decreased insulin response to a glucose bolus (300 mg/kg i.v.). Pretreatment with the peripherally acting 5-HT1c/5-HT2 receptor antagonist, BW 501C67 (0.5 mg/kg i.v. 10 min beforehand) prevented the rise in plasma glucose triggered by the peripherally acting 5-HT1c/5-HT2 receptor agonist, alpha-methyl-5-HT (0.75 mg/kg i.v.), but amplified the rise elicited by DOI. Pretreatment with chlorisondamine (1 mg/kg i.v. 10 min beforehand) or adrenalectomy 20 h beforehand prevented the DOI-induced hyperglycemia. On the other hand, pretreatment with dexamethasone (0.35 mg/kg s.c. 2 h and 20 min beforehand) did not affect the DOI-induced hyperglycemia. It is concluded that the hyperglycemic effect of DOI administration is mediated by centrally located 5-HT2 receptors and, in turn, adrenal epinephrine release.


Characterization of muscarinic receptors mediating vasodilation in guinea-pig ileum submucosal arterioles by the use of computer-assisted videomicroscopy. Muscarinic receptors of resistance vessels (submucosal arterioles, outside diameter 50-75 microns) from the guinea-pig small intestine were investigated in vitro using a computer-assisted videomicroscopy system (Diamtrak). The muscarinic receptor which mediates vasodilation of precontracted [U-46619 (300 nM) or (-)-noradrenaline (10 microM)] arterioles was characterized with several muscarinic agonists and subtype-selective antagonists. The following agonists all produced equivalent maximum vasodilation (given in rank order of potency): acetylcholine = arecaidine propargyl ester (APE) greater than oxotremorine = (+/-)-muscarine = (+/-)-methacholine greater than carbachol greater than 4-[[N-(4-chlorophenyl)carbamoyl]oxy]-2-butynyltrimethylammonium iodide (4-Cl-McN-A-343). 4-[[N-(3-Chlorophenyl)-carbamoyl]oxy]-2-butynyltrimethylammonium chloride (McN-A-343) and N-ethyl-guvacine propargyl ester (NEN-APE) produced minimal or no arteriolar vasodilation. The muscarinic antagonists pirenzepine, (+-)-5,11-dihydro-11-[[[2-[2-((dipropylamino)methyl)-1-piperidinyl] ethyl]amino]-carbonyl]-6H-pyrido(2,3-b)(1,4)-benzodiazepin-6-one (AF-DX 384), 11-[[4-[4-(diethylamino)butyl]-1-piperidinyl]acetyl]-5,11-dihydro- 6H-pyrido(2,3-b)(1,4)-benzodiazepin-6-one (AQ-RA 741), p-fluorohexahydro-sila-difenidol (p-F-HHSiD), 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP) and (R)- and (S)-hexahydro-difenidol [(R)-HHD, (S)-HHD] shifted the muscarine, methacholine or carbachol dose-response curve to the right in a competitive manner. Schild analysis of the data yielded pA2 values for pirenzepine (6.74/6.9), AF-DX 384 (6.72), AQ-RA 741 (6.58), p-F-HHSiD (7.53/7.57), 4-DAMP (9.06), (R)-HHD (7.88/8.32) and (S)-HHD (5.52/5.88). Thus, it can be concluded that submucosal arterioles possess only the M3 functional muscarinic receptor, the activation of which causes blood vessel dilation.(ABSTRACT TRUNCATED AT 250 WORDS)


Excitatory effects of adenosine 5'-triphosphate on rat locus coeruleus neurones. Pontine slices of the rat brain were used for extracellular recording of the frequency of spontaneous action potentials of locus coeruleus (LC) neurones. In the absence of 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), alpha,beta-methyleneadenosine 5'-triphosphate (alpha,beta-meATP; 0.3-30 mumol/l) and 2-methylthio ATP (0.3-100 mumol/l), but not ATP (1-100 mumol/l) increased the firing rate. In the presence of DPCPX 0.1 mumol/l, all three purinoceptor agonists were active, the potency order being alpha,beta-meATP greater than 2-methylthio ATP = ATP. Preincubation of the slices with tetrodotoxin (TTX) 0.5 mumol/l decreased the spike discharge but did not alter the percent facilitatory effect of alpha,beta-meATP 30 mumol/l. There was no desensitization to alpha,beta-meATP 10 mumol/l on repeated or continuous application. Suramin 100 mumol/l selectively depressed the effect of alpha,beta-meATP 30 mumol/l without interfering with the effect of equiactive concentrations (10-100 mumol/l) of glutamic acid. The concentration-response curve of alpha,beta-meATP was shifted in a parallel manner to the right by suramin 10 mumol/l. While DPCPX 0.1 mumol/l facilitated firing, suramin 100 mumol/l did not change it. In conclusion, LC neurones may possess P2-purinoceptors of an unidentified type, which share some P2x characteristics.


Naltrindole retards tolerance development to morphine-induced effects on EEG and EEG power spectra. In the present study, EEG and EEG power spectra were used to assess the effects of naltrindole, a selective delta opioid antagonist, on the development of tolerance to morphine. Adult female Sprague-Dawley rats were implanted with cortical EEG electrodes and permanent indwelling i.c.v. and i.v. cannulas. Twice daily for 7 days, rats were pretreated with either i.c.v. naltrindole (20 nmol) or i.c.v. water, 20 min before i.v. morphine (10 mg/kg) injections. The treatments produced EEG slow-wave bursts and associated behavioral stupor. The amount and duration of these effects decreased less rapidly over the 7 days in the naltrindole-pretreated rats than in the water-pretreated rats. I.c.v. naltrindole pretreatment also prevented significant decreases in latency to onset of slow-wave sleep that were seen in the i.c.v. water-pretreated group. EEG data were further analyzed on a Pathfinder II computer. The development of tolerance was reflected by decreases in the total absolute EEG spectral power (1-50 Hz) over the 7-day period. Rats that were pretreated with i.c.v. naltrindole (20 nmol) did not display a significant decrease in total absolute EEG spectral power by the 7th day, as did the i.c.v. water-pretreated group. Furthermore, significant differences were seen for complexity, mobility, and edge frequency between the two pretreatment groups. A delayed qualitative change in the EEG power spectra was also observed in rats pretreated with i.c.v. naltrindole. On day 1, EEG slow-wave bursts were associated with increases in EEG spectral power over the 1-10 Hz range.(ABSTRACT TRUNCATED AT 250 WORDS)


Effects of nor-binaltorphimine on the development of analgesic tolerance to and physical dependence on morphine. The effects of a highly selective kappa antagonist, nor-binaltorphimine (nor-BNI), on the development of tolerance to morphine analgesia and physical dependence on morphine were examined. Pretreatment with nor-BNI (5 mg/kg s.c.) 2 h prior to injection of morphine or a selective kappa agonist, U-50,488H, significantly antagonized the analgesic effect of U-50,488H, but not morphine analgesia in mice. The development of tolerance to morphine analgesia was significantly potentiated by pretreatment of mice with nor-BNI 2 h prior to morphine treatment during chronic morphine treatment for 5 days. Additionally, the pretreatment with nor-BNI during chronic treatment with the high dose of morphine for 5 days significantly potentiated the naloxone-induced body weight loss in morphine-dependent mice and rats. These findings suggest that inactivation of the kappa opioid system may potentiate the development of tolerance to morphine analgesia in mice and may aggravate the naloxone-precipitated body weight loss in morphine-dependent mice and rats.


Effects of cocaine and its quaternary derivative cocaine methiodide on cardiovascular function in squirrel monkeys. The effects of cocaine and its quaternary derivative cocaine methiodide, which does not cross the blood-brain barrier, were studied on cardiovascular function in squirrel monkeys. In conscious monkeys, cocaine produced clear dose-dependent increases in blood pressure and heart rate, while cocaine methiodide did not. Both cocaine and cocaine methiodide enhanced the effects of norepinephrine in anesthetized animals, suggesting that both inhibit neuronal uptake of norepinephrine; cocaine was approximately 30 times more potent than cocaine methiodide. In anesthetized monkeys both cocaine and cocaine methiodide produced small, short duration pressor effects, although cocaine was at least 10 times more potent than cocaine methiodide. Cocaine's effects in anesthetized animals were clearly blunted in comparison to its effects in conscious animals. These effects of cocaine on blood pressure occurred at doses lower than those required to enhance norepinephrine's effects, indicating that the norepinephrine uptake blocking effects of the drugs cannot fully account for their cardiovascular effects. The greatly enhanced effect of cocaine in conscious animals and the finding that cocaine methiodide had little effect in conscious animals indicates that central mechanisms are involved in the effects of cocaine on cardiovascular function in conscious animals.


Effects of methoxamine and barium on 45Ca2+ fluxes in the rat vas deferens. The aim of this study was to determine whether methoxamine and barium stimulate 45Ca2+ uptake or efflux in the rat vas deferens in a manner that correlates with their contractile activity, and whether 45Ca2+ movements are inhibited by verapamil or nifedipine. Basal La(3+)-resistant (cellular) 45Ca2+ uptake was significantly greater in the epididymal half (791 +/- 27 nmol g-1) than in the prostatic half (654 +/- 14 nmol g-1) of the rat vas deferens and was unaffected by verapamil (61 microM) or nifedipine (14 microM). Methoxamine (8 microM) was without effect on 45Ca2+ uptake in either half but BaCl2 (1 mM) increased 45Ca2+ uptake by 31% in the prostatic half and by 22% in the epididymal half. The barium-induced increases in 45Ca2+ uptake were markedly reduced or abolished by verapamil (2 microM) or nifedipine (0.3 microM), which at these concentrations have no effect on the rhythmic contractions but abolish the initial small phasic contraction induced by barium. The basal rate of 45Ca2+ efflux from the intact vas deferens (into Ca2+ containing Krebs-Henseleit solution or into Ca-free Krebs-Henseleit solution +/- EGTA 0.05 mM) was not affected by verapamil (61 microM) or nifedipine (14 microM). Methoxamine (8 microM) produced a marked, transient and reversible increase in 45Ca2+ efflux into 2.5 mM CaCl2 Krebs-Henseleit in 50% of the intact vasa deferentia examined which was augmented by verapamil (61 microM). BaCl2 (1 mM) produced a small increase in 45Ca2+ efflux into Ca(2+)-containing and Ca(2+)-free Krebs-Henseleit solutions from some intact vasa deferentia and this was not inhibited by nifedipine (14 microM).(ABSTRACT TRUNCATED AT 250 WORDS)


Prolactin induces growth-related gene expression in rat aortic smooth muscle in vivo. We examined the effects of in vivo administration of prolactin on growth-related gene expression in aorta. Optimal mRNA expression for both the proto-oncogene, c-myc and ornithine decarboxylase occurred at 22 mg/kg prolactin. For c-myc, this was seen as early as 15 min. Prolactin-induced ornithine decarboxylase mRNA expression began at 1 h. The results confirm work showing induction of these genes by prolactin in tissues where it is mitogenic and supports a role for prolactin in the trophic response of vascular smooth muscle.


Transferable antibiotic resistance among thermotolerant coliforms from rural drinking water in India. A total of 231 thermotolerant coliforms was isolated from rural drinking water from four states of India. Of these, 220 isolates were resistant to ampicillin, chloramphemicol, streptomycin and tetracycline. Multiple (MAR), double and single antibiotic resistances were observed in 31.4, 48.6 and 13.7% of the isolates, respectively. Out of 177 antibiotic-resistant isolates examined for transmissibility, only 15.3% were able to transfer their resistances to Escherichia coli K-12 recipient. The resistances were transferred by 32.5% of MAR, 21.9% of double resistant and 7.6% of single resistant isolates. Ampicillin resistance was transferable in 14.69% strains while resistances for the rest of the antibiotics were transferable in less than 4% strains. MAR strains of E. coli and Klebsiella sp. showed highest levels of R-plasmid transfer.


Giardia--diagnosis, clinical course and epidemiology. A review. Infection with giardia may be associated with significant ill-health and while the reported incidence of infection is increasing in the United Kingdom, the true prevalence of infection and extent of morbidity due to this organism is unknown. Diagnosis is made difficult by non-specificity of symptoms and low sensitivity of traditional diagnostic techniques. Immunological methods of diagnosis hold promise for the future, but in the meantime, more routine testing by laboratories and multiple faecal testing by clinicians may prevent unnecessary morbidity. The late summer/autumn peak in reported infection is difficult to explain while the age distribution is typical of an organism which is spread faeco-orally. The importance of potable water supplies as a source of infection in this country is not clear, nor is the role of zoonotic spread. The apparent susceptibility to infection of certain population groups requires further exploration as does the role of the asymptomatically infected in transmission.


The ability of airborne Klebsiella pneumoniae to colonize mouse lungs. A strain of Klebsiella pneumoniae was aerosolized and its survival in air at different relative humidities was studied. Survival was dependent upon relative humidity and aerosols were most stable during storage at a relative humidity of 60%. Mice were exposed to aerosols of K. pneumoniae produced at this humidity and lung samples taken at timed intervals after exposure. Fifteen strains of K. pneumoniae were tested for their ability to colonize mice, but only five were detectable in mouse lungs 7 days after exposure. Three of these strains persisted without an increase in bacterial numbers, regardless of the initial inoculum used. Two strains of K. pneumoniae, designated strains 15 and 16, persisted in a similar manner when used at a low dose; however, when the dose received per lung was increased there was a rapid multiplication of bacteria in the lungs.