Cross-talk between excitatory and inhibitory amino acids in the regulation of luteinizing hormone-releasing hormone secretion. Inhibitory (IAA) and excitatory amino acid (EAA) neurotransmitters appear to play an important role in regulating reproductive functions. L-Glutamic acid (GLU), the major representative of the EAA system, stimulates LHRH release from arcuate nucleus-median eminence (AN-ME) fragments in vitro. Several studies have provided evidence for considering gamma-aminobutyric acid (GABA), a major IAA neurotransmitter, as another regulator of LHRH secretion. Recent reports have indicated that a cross-talk between GABA and GLU participates in the regulation of synaptic transmission in the brain. In concert with this notion, we present evidence indicating that this cross-talk between GABA and GLU appears to be also involved in neuroendocrinological paradigms. In this respect, bicuculline, a GABA-A receptor antagonist, blocked GLU-evoked LHRH secretion from AN-ME fragments in vitro without affecting basal LHRH release. In addition, activation of GABA-A receptors by muscimol (MUS) stimulated basal LHRH secretion. Interestingly, when MUS and GLU were added together to the incubation medium, an additive, stimulatory effect was observed. These observations clearly indicate that a GABAergic mechanism participates, via GABA-A receptors, in GLU-induced LHRH secretion from terminals of the ME. Furthermore, GABA-B receptors appear to negatively modulate the effects of GLU. Activation of GABA-B receptors by baclofen (BAC) blocked GLU-induced LHRH secretion, while phaclofen, a GABA-B receptor antagonist, reversed this effect. In summary, our data provide evidence for a cross-talk between EAA and IAA systems in the regulation of LHRH release, and, therefore, in the control of gonadal function.


Errors in sizing bands of hypervariable DNA profiles on autoradiograms: are they Gaussian? The accuracy of procedures for sizing hypervariable restriction fragments by Southern blot analysis (SBA) has been tested under three different experimental conditions: (i) intrablot serial analyses: three heterozygous DNA profiles were tested 14 times each in the same gel electrophoresis; (ii) intralaboratory analyses: we replicated three profiles (six autoradiographic bands) in over 100 SBA experiments; (iii) interlaboratory analyses: 15 serial measurements produced in a recent collaborative study (Forensic Sci. Int. 1991, 49, 1-15) were taken into account. In these three cases, a typical U-shaped correlation curve between molecular size and coefficient of variation was found. We explain decrease of accuracy at both extremities of the gels in terms of: (i) enlarged shapes of bands and mean electrophoretic resolution at the cathode; (ii) diffusion and blurring of bands at the anodal edge. The three populations of data were subjected to a chi 2 test and to the Kolmogorov-Smirnov test in order to verify their compliance with normal distribution. Twenty-three out of 27 tests indicated no significant deviation from the assumption of Gaussian distribution. We recommend the adoption of tests for normality to validate the use of symmetric confidence intervals for calculating gene frequencies and asserting a match between adjacent bands.


Deletion studies to reveal the basis for size discrepancy in proliferating cell nuclear antigen. Proliferating cell nuclear antigen (PCNA), an essential component for DNA replication in eukaryotes, is a highly conserved nonhistone nuclear protein of 261 amino acids. The molecular weight of mammalian PCNA, estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), differs notably from that predicted by the cDNA sequences, that is, 36,000 in comparison with 29,261 and 28,748 for human and rat PCNA, respectively. To investigate if this discrepancy is due to posttranslational modifications, we studied the PCNA protein synthesized by an in vitro transcription/translation system as well as the protein overproduced in bacteria. We found that both PCNA protein samples were indistinguishable from the authentic protein from the protein mobility in SDS-PAGE. The finding indicates that the size discrepancy is not due to the posttranslational modifications. Hence, the size discrepancy may be due to the protein sequence per se, namely a sequence-related anomaly in SDS-PAGE. Results from the analyses of a series of PCNA derivatives with various lengths of C- or N-terminal deletion indicate that the putative sequence is in the region of residues 128-150.


v-erbB oncogene expression accounts for most variations in protein synthesis after avian erythroblastosis virus infection of chicken embryo fibroblasts: a two-dimensional electrophoresis study. The effect of the v-erbA and/or v-erbB oncogenes on cellular gene expression was investigated after separation by two-dimensional polyacrylamide gel electrophoresis of [35S]methionine-labelled proteins from chicken embryo fibroblasts (CEF), infected by either the avian erythroblastosis virus (AEV) carrying both oncogenes, or by viruses carrying only one of them. We observed significant changes in the synthesis of 34 proteins in AEV-transformed CEF as compared with control cells. The synthesis of 24 of them was increased while the synthesis of the other 10 proteins was decreased. The expression of v-erbB alone is necessary and sufficient to induce changes in the synthesis of 27 proteins while the 7 remaining modifications are observed only in cells expressing v-erbB together with v-erbA. Moreover, the deregulation of protein synthesis by v-erbB-expressing viruses was correlated with the morphological transformation state of cells.


Common accessory genes for the Bordetella pertussis filamentous hemagglutinin and fimbriae share sequence similarities with the papC and papD gene families. The Bordetella pertussis filamentous hemagglutinin (FHA) is a major virulence factor responsible for attachment, one of the early events in bacterial pathogenesis. Deletion of its structural gene, fhaB, or a Tn5 insertion in fhaA, downstream of fhaB, resulted in a FHA- and fimbriae- phenotype, although fhaB and the fim genes are not linked. The fhaB downstream region therefore most likely encodes accessory proteins required for the biosynthesis of FHA and fimbriae, despite the lack of sequence similarities between these two proteins. The nucleotide sequence of this area contains the open reading frames fhaD and fhaA, whose products share sequence similarities with the papD and papC gene products, respectively. PapD is a periplasmic chaperone protein able to bind to the Escherichia coli P pilin subunits and to transport them towards the outer membrane protein PapC which is responsible for pilus membrane translocation. An additional open reading frame, fhaE, is located downstream of fhaA. Its amino acid sequence shares similarities with those of the fimbrial subunits. Deletion analyses suggest that fhaB and the downstream genes can be transcribed as a polycistronic operon, and primer extension analysis revealed the presence of a second promoter between fhaB and fhaD.


Phosphorylation of CREB affects its binding to high and low affinity sites: implications for cAMP induced gene transcription. Cyclic AMP treatment of hepatoma cells leads to increased protein binding at the cyclic AMP response element (CRE) of the tyrosine aminotransferase (TAT) gene in vivo, as revealed by genomic footprinting, whereas no increase is observed at the CRE of the phosphoenolpyruvate carboxykinase (PEPCK) gene. Several criteria establish that the 43 kDa CREB protein is interacting with both of these sites. Two classes of CRE with different affinity for CREB are described. One class, including the TATCRE, is characterized by asymmetric and weak binding sites (CGTCA), whereas the second class containing symmetrical TGACGTCA sites shows a much higher binding affinity for CREB. Both classes show an increase in binding after phosphorylation of CREB by protein kinase A (PKA). An in vivo phosphorylation-dependent change in binding of CREB increases the occupancy of weak binding sites used for transactivation, such as the TATCRE, while high affinity sites may have constitutive binding of transcriptionally active and inactive CREB dimers, as demonstrated by in vivo footprinting at the PEPCK CRE. Thus, lower basal level and higher relative stimulation of transcription by cyclic AMP through low affinity CREs should result, allowing finely tuned control of gene activation.


Transcriptional repression of band 3 and CAII in v-erbA transformed erythroblasts accounts for an important part of the leukaemic phenotype. The v-erbA oncogene confers two prominent properties on transformed erythroblasts: a block of spontaneous differentiation and tolerance to wide variations in the pH or ionic strength of culture medium. V-erbA acts as a constitutive repressor of erythrocyte-specific gene transcription, arresting the expression of at least three different erythroid genes: the erythrocyte anion transporter (band 3), carbonic anhydrase II (CAII) and delta-aminolevulinate synthase (ALA-S). To test whether or not the v-erbA induced repression of these genes is causally related to the v-erbA induced leukaemic phenotype, we have reintroduced the genes for band 3 or CAII into transformed erythroblasts via retrovirus vectors. We show here that such erythroblasts, expressing v-erbA, require the same narrow range of medium pH and ion concentration for growth as do transformed erythroblasts lacking v-erbA, i.e. the v-erbA induced tolerance to pH variation was abrogated. The v-erbA induced differentiation block, however, remained unaffected by the re-expression of band 3 and was only slightly affected by the re-expression of CAII. Our experiments show that the two v-erbA-related 'erythroblast transformation parameters' are separable: suppression of band 3 and CAII accounts for one parameter (pH/ion tolerance), while the second parameter (differentiation block) must involve v-erbA regulation of a different set of target genes.


Zmhox1a, the product of a novel maize homeobox gene, interacts with the Shrunken 26 bp feedback control element. A new maize homeobox gene was isolated by screening a lambda gt11 expression library with the 26 bp Shrunken feedback control element. Zmhox1a (Zea mays homeobox) is an unidentified maize gene mapping to the long arm of chromosome 8. It is a member of a new class of maize homeobox genes only distantly related to the Knotted class. The 3.1 kb Zmhox1a transcript can be detected in different maize tissues and encodes a polypeptide of 719 amino acids. Western blotting experiments detect the native 112 or 115 kDa protein in nuclear protein extracts, the nuclear localization being compatible with a function in transcriptional control. No Zmhox1a protein is detected in maize roots despite the presence of the Zmhox1a transcript; this may indicate a post-transcriptional control mechanism. A highly acidic central region of the Zmhox1a polypeptide implies a transcriptional activator function. The carboxy-terminal part of the maize homeodomain protein is related to the human Oct2 transcription factor, but homology to the POU specific domain is restricted to the POU-B subdomain. It was confirmed by DNase I footprinting experiments that DNA binding of the Zmhox1a homeodomain was at three sites flanking the TATA-box of the Shrunken promoter.


Plasma endogenous opioid levels in acute myocardial infarction patients, with and without pain. Plasma levels of beta-endorphin, met-enkephalin and dynorphin were assessed in acute myocardial infarction (AMI) patients, with and without pain (group I: no pain, N = 12; group II: severe pain, N = 16). Plasma opioid peptide concentration was measured on admission to hospital (between 1 and 3 h after the myocardial infarction onset), at 7, 12, 24 h and at 2, 3 and 4 days. A transient increase in plasma beta-endorphin levels was found in AMI patients with severe pain, the levels normalizing within 12-18 h when pain had ceased. No changes in beta-endorphin concentration were observed in AMI patients without pain. Compared with healthy subjects, low levels of met-enkephalin were found in both groups of AMI patients throughout the study. Low levels of dynorphin were observed in patients with no pain while in the other patients initial low levels of dynorphin normalized when pain ceased. Blood pressure, heart rate and central venous pressure values were normal and did not correlate with plasma opioid levels. The results suggest that endogenous opioids do not affect pain in the early phase of myocardial infarction. The rise in beta-endorphin concentration observed in patients with severe pain seems to be induced by pain stress.


Comparative effects of epanolol and diltiazem on exercise performance and respiratory gas exchange in angina pectoris. The effects of epanolol (a new selective beta-adrenoceptor antagonist), diltiazem and placebo were compared in a group of 16 patients with chronic stable angina pectoris. Each patient received each treatment in random order. Diltiazem reduced weekly angina attack rate from 7.2 (95% CI 3.9-10.5) to 3.9 (1.9-5.9) (P less than 0.01), whereas a lesser reduction was observed after epanolol. Both drugs produced a small but significant (P less than 0.05) increase in treadmill exercise time (placebo 474 s (374-574), epanolol 527 s (431-623) and diltiazem 554 s (462-646). However, aerobic work capacity, assessed by peak achieved oxygen consumption, was not different from the placebo value of 21.2 (18.0-24.4) ml.min-1.kg-1, and clearly subnormal when compared to age- and sex-matched controls (33.0 (30.1-35.9) ml.min-1.kg-1). Ventilatory abnormalities and increased lactate levels on active treatment were observed at peak exercise only. We conclude that the cardiodepressant effects of both active drugs limit blood supply to working skeletal muscle, and that chest pain may be replaced by dyspnoea or fatigue as the limiting factors to exercise.


Effects of mental and physical stress on plasma catecholamine levels before and after beta-adrenoceptor blocker treatment. In a study in mild hypertensives, the impact of mental and physical stress on plasma epinephrine (E), norepinephrine (NE), and on their ratio (NE/E) was evaluated. The effect of two beta-adrenoceptor blocking drugs, atenolol and bopindolol, on plasma catecholamine levels was also examined. Each stressful stimulus significantly increased the NE and E levels compared to rest. The increase was progressive from mental stress, through the handgrip test to the treadmill test. A slight decrease in the NE/E ratio was observed following mental stress and the handgrip test, while this ratio increased during the treadmill test. No significant impact of beta blocking treatment on catecholamine levels was observed under any test condition.


Peripheral hematopoietic stem cell transplantation: current concepts. As methods for increasing stem cells are perfected and alternate regimens for transplantation developed, PSCT will undoubtedly see wider application in combination with BMT and may ultimately replace BMT. The initial encouraging results with PSCT so far portend a major therapeutic role of this modality in the approach to hematologic and oncologic diseases. Prospective randomized trials comparing PSCT and BMT in a variety of clinical settings are needed and are already underway.


Acute effect of an alpha 1-adrenoceptor antagonist on urinary sodium excretion, plasma atrial natriuretic peptide, arginine vasopressin, and the renin-aldosterone system in healthy subjects. To elucidate the mechanism underlying the sodium retention caused by alpha 1-adrenoceptor blockade in man, a placebo-controlled, randomised, double-blind study has been made of the acute effects of bunazosin an alpha 1-antagonist, on urinary sodium excretion, atrial natriuretic peptide (ANP), arginine vasopressin (AVP), and the renin-aldosterone system in 7 healthy men. A single oral dose of bunazosin 2.0 mg caused a significant reduction (P less than 0.05) in urinary sodium excretion after 0-2 h, 2-4 h, and 4-6 h. The mean values for plasma ANP, AVP, aldosterone, and cortisol concentrations at those times were similar after placebo and bunazosin, and plasma renin activity was significantly increased 2 and 4 h after bunazosin. Pretreatment with oral enalapril 10 mg, an angiotensin converting enzyme inhibitor, did not prevent the bunazosin-induced reduction in urinary sodium excretion. There was a significant positive correlation between the drug-induced changes in blood pressure and urinary sodium excretion. The results suggest that ANP, AVP, and renin-aldosterone may play little role in the sodium retention caused by acute alpha 1-adrenoceptor blockade in man.


Effects of escalating doses of recombinant human interleukin-2 in correcting functional T-cell defects following autologous bone marrow transplantation for lymphomas and solid tumors. High-dose chemotherapy followed by autologous bone marrow transplantation (ABMT) in the treatment of malignancies is often associated with immune deficiency following transplantation, possibly contributing to tumor relapse or fatal infection. Interleukin 2 (IL-2), which enhances major histocompatibility complex (MHC)-unrestricted cytotoxicity in vitro, can be applied in vivo as immunotherapy to reduce these potential complications. Recombinant human IL-2 (rIL-2) was administered by continuous i.v. infusion for four courses in 19 patients following ABMT for lymphomas and solid tumors. The patients were assigned to five groups of escalating doses of rIL-2 ranging from 3 to 30 x 10(6) IU/m2/day. The immunological effects and toxicity were monitored. After a transient reduction of lymphocytes in the peripheral blood, a significant lymphocytosis was observed during the rIL-2 infusion with an augmentation of CD2+, CD25+, and CD8(+)-Ia+ T cells and a dose-related increase of CD56+ lymphocytes. Natural killer (NK) activity appeared enhanced in patients treated with as little as 6 x 10(6) IU/m2/day. No statistically significant increase in lymphokine-activated killer (LAK) activity was seen after rIL-2, when compared to LAK activity following ABMT prior to rIL-2 administration. Administration of exogenous rIL-2 to patients who have undergone ablative chemotherapy and ABMT has a role in restoring defective T-cell function. Further trials defining those patients most likely to benefit from rIL-2 integrated with ablative chemotherapy and ABMT are now warranted.


RFLPs of the LDL-receptor gene: their use in the diagnosis of FH and in evaluation of different levels of gene expression on normal subjects. The usefulness of the RFLPs of the LDL-receptor gene in early diagnosis of Familial Hypercholesterolemia (FH) was investigated in 122 FH-families. Four RFLPs, produced by digestion with the enzymes PvuII, ApaLI and AvaII/XbaI were able to detect the affected gene and to follow the inheritance of the disease in 72 out of 97 families (74%). In the remaining 25 families, unambiguous diagnosis was possible in 66% of the cases by use of PvuII, ApaLI and BstEII/EcoRI RFLPs. The RFLPs were also useful to distinguish true homozygotes from compound heterozygotes and to detect families where recombination events occurred or where hypercholesterolemia was not due to a defect of the LDL-receptor gene. In a normal population PvuII RFLP account for 9.6% of the total variance of the LDL cholesterol levels adjusted for confounding variables. The P2 allele was associated with lower LDL cholesterol concentrations (average excess -9.1 mg/dl). This finding allows us to presume there is a DNA sequence, close to the variable PvuII cutting site in intron 15, which could act as an enhancer of the LDL-receptor gene expression.


Application of cladistics to the analysis of genotype-phenotype relationships. We seek to understand the relative contribution of allelic variations of a particular gene to the determination of an individual's risk of atherosclerosis or hypertension. Work in progress is focusing on the identification and characterization of mutations in candidate genes that are known to be involved in determining the phenotypic expression of intermediate biochemical and physiological traits that are in the pathway of causation between genetic variation and variation in risk of disease. The statistical strategy described in this paper is designed to aid geneticists and molecular biologists in their search to find the DNA sequences responsible for the genetic component of variation in these traits. With this information we will have a more complete understanding of the nature of the organization of the genetic variation responsible for quantitative variation in risk of disease. It will then be possible to fully evaluate the utility of measured genetic information in predicting the risk of common diseases having a complex multifactorial etiology, such as atherosclerosis and hypertension.


Apo C-II deficiency type Bari.

Apo C-II deficiency type Bari. We formerly studied an Italian family with apo C-II deficiency. Two probands were homozygous for the defect (unmeasurable circulating apolipoprotein C-II and absence of C-II bands on immunoelectrophoresis). We documented the synthesis of the protein at the intestinal level in the probands with immunohistological techniques. With the purpose of investigating the molecular basis of the defect, Southern analysis, polymerase chain reaction (PCR) amplification and sequence analysis were carried out on one of the two cases. We identified a point mutation C to G transversion in the third exon of the gene causing a premature stop codon. Our hypothesis is that the truncated protein of 36 aa., instead of 79 aa., lacks its functional domain. This causes inefficiency in the activation of lipoprotein lipase (LPL) and the instability of the circulating molecule, which could have an higher catabolic rate compared to a normal protein. The faster disappearance from the circulating compartment make it unmeasurable. The mutation destroys a Rsa I site, present in the normal gene sequence. We suggest the use of this site for a rapid Restriction Fragment Length Polymorphism (RFLP) on PCR amplification products to screen this defect in the Italian population.


Streptomyces lividans possesses a GroEL-like chaperonin. Streptomyces lividans grown at 45 degrees C produces a GroEL-like chaperonin. This protein is specifically synthesized in bacterial cell cultures upon heat shock induction. It has a similar size (62 kDa) to the GroEL-like proteins from Escherichia coli and Bacillus subtilus and shows immunological cross-reaction with serum raised against GroEL from E. coli. The S. lividans 62-kDa protein assembles into oligomers around 20S that show a morphology consistent with a barrel showing six-fold and seven-fold symmetries as previously described in E. coli and B. subtilis.


[Comparison of the effects of benzodiazepine and non-benzodiazepine anxiolytics on agonistic behavior in male mice]. The present study investigated whether there is any difference between the effects of benzodiazepine and non-benzodiazepine anxiolytics on agonistic behavior in male mice, using an ethopharmacological technique. Agonistic behavior was evoked using a resident-intruder paradigm. The effects of four doses of the following drugs were assessed in either resident or intruder mice: diazepam (vehicle, 1, 2.5 and 5 mg/kg, p.o.) and tandospirone (vehicle, 2.5, 5 and 10 mg/kg, p.o.). Residents and intruders were drugged on alternate test days, and all animals received different sequences of each of the drug conditions according to a random schedule. The injection-test interval was 30 min. When a resident mice were treated with either diazepam or tandospirone, the frequency of attack bite was suppressed significantly in a dose-dependent manner. When intruder mice were treated with diazepam, attack bites by untreated residents were significantly increased, whereas tandospirone was ineffective. Although diazepam caused a significant decrease in both locomotion and rearing, tandospirone did not cause motor dysfunction. These evidence indicate that tandospirone, a 5-HT1A receptor agonist, has different pharmacological properties from diazepam.


[Effects of IGN-2098, a new histamine H2-receptor antagonist, on gastric secretion and gastric and duodenal lesions induced in rats. Comparison with roxatidine]. A new compound, IGN-2098 [5,6-dimethyl-2-[4-<3-(1-piperidinomethyl) phenoxy>cis-butenylamino]-4-(1H)-pyrimidone.2HCl], was found to be a potential histamine H2-receptor antagonist in the guinea pig atrium. IGN-2098, given p.o., significantly and persistently (for more than 12 hr) inhibited the basal gastric secretion in pylorus-ligated rats. The agent also significantly inhibited the basal gastric secretion when given by the s.c.-, i.d.- or i.p.-route. Stimulated gastric secretion in fistula rats in response to histamine, carbachol or pentagastrin was also significantly inhibited with IGN-2098 given s.c. Pretreatment with IGN-2098 (p.o.) significantly protected the gastric mucosa against pylorus ligation-, water-immersion stress-, histamine-, indomethacin-, HCl.aspirin-, and HCl.ethanol-induced gastric lesions. In addition, the agent significantly protected the duodenal mucosa against mepirizole-induced ulcers. Based upon the ED50 values, the antisecretory effects on histamine, carbachol or pentagastrin-stimulated acid secretion were 6.0, 37.0 or 80 times more potent than roxatidine, respectively. As to the anti-lesion effects on HCl.aspirin-induced gastric lesions or mepirizole-induced duodenal ulcers, IGN-2098 was 8.1 or 14.8 times more potent than roxatidine, respectively. These results suggest that IGN-2098 will be a useful drug for the treatment of gastric and duodenal lesions in man.


Latex hypersensitivity reactions despite prophylaxis. Latex rubber hypersensitivity represents a significant problem facing the medical, surgical, radiologic, and dental professions. As a tertiary care center, the Childrens Hospital of Philadelphia has a large population of patients with spina bifida and complex genitourinary anomalies; a number of these children have latex rubber allergy, which may first present as intraoperative anaphylaxis. Although there is no substitute for complete antigen avoidance, all medical products containing latex rubber may not have suitable alternatives. Therefore, we have formulated a protocol to prevent perioperative reactions through the use of prophylactic medications and the limitation of latex exposure. This regimen includes steroids, antihistamines, and bronchodilators when indicated. In four children, prophylaxis failed perioperatively because of parenteral infusion of latex rubber proteins.


Importance of imidazoline receptors in the cardiovascular responses to clonidine and rilmenidine in conscious rabbits. The present paper summarizes our studies concerning the involvement of imidazoline and alpha 2-adrenoceptors in the cardiovascular actions of centrally acting drugs rilmenidine, clonidine and methyldopa. We have found that they produce very similar cardiovascular autonomic effects which relate directly to the function of central monoamine neurotransmitters. They mimic certain elements of the noradrenergic neuron system in the central nervous system, in particular the brainstem actions which involve hypotension, bradycardia and resetting of the baroreceptor heart rate reflex. By contrast they turn off serotonergic pathways that are pressor, produce tachycardia and inhibit the baroreceptor heart rate reflex. Recent studies using specific receptors antagonist drugs idazoxan and 2-methoxy-idazoxan indicate that in conscious rabbits the imidazoline receptor actions of rilmenidine is of primary importance at doses which would be considered clinically relevant. We further conclude that the alpha 2-adrenoceptors and the imidazoline receptors are likely to be located in series i.e. along the same cardiovascular autonomic pathways in the brainstem but presumably at different sites.


Individualized treatment of duodenal ulcer disease. A pilot study. A substantial number of duodenal ulcer (DU) patients relapse despite maintenance treatment with antisecretory drugs. The influence of certain risk factors and the heterogeneity of the disease could explain such behavior. The present prospective, open study compares the one-year clinical outcome (with upper GI endoscopy at the beginning of the study, at 6 and 12 months, and at every symptomatic relapse) of four groups of DU subjects, consecutively recruited from December 1987 to December 1988, separated in accordance with whether or not a bleeding DU episode had previously occurred, and whether or not an evaluation of gastric acid secretion had been made. Thus, Group I (17 patients; 12 males, 5 females) included heavy smokers and/or gastric acid hypersecretors; Group II (13 patients; 12 males, 1 female) non- or light smokers non-hypersecretors; Group III (34 patients; 22 males, 12 females) subjects with unknown gastric acid secretion; Group IV (33 patients; 30 males, 3 females) previously bleeding DU patients. All patients, except those in Group II (who were left untreated), were given ranitidine 150 mg at bedtime. The outcome of Groups I+II was compared with that of Group III (considered as "standard therapy") and Group IV patients, the latter presumably with a low risk of relapse because of the low prevalence of smokers. Chi-square test, Fisher's exact test, analysis of variance and the logrank test. During the year of follow-up, 27/97 patients withdrew from the study, while 18 had a DU relapse (remission rates 82.1% +/- 7.4% in Groups I+II, 70.5% +/- 8.4% in Group III, 87.5% +/- 5.9% in Group IV).(ABSTRACT TRUNCATED AT 250 WORDS)


Detection of circulating intercellular adhesion molecule-1 in chronic liver diseases. The leucocyte adhesion molecule intercellular adhesion molecule-1 is induced on bile ducts in patients with primary biliary cirrhosis and primary sclerosing cholangitis and may be involved in targeting immune damage to these structures. It has recently been reported that, when activated, in vitro lymphocytes release a soluble form of intercellular adhesion molecule-1 that can also be detected in human serum. Because it is functionally active, this circulating intercellular adhesion molecule-1 might play a role in regulating inflammation by blocking adhesion. We used an enzyme-linked immunosorbent assay to detect circulating intercellular adhesion molecule-1 in the serum of patients with primary biliary cirrhosis and primary sclerosing cholangitis. Levels of circulating intercellular adhesion molecule-1 were markedly elevated in primary biliary cirrhosis and primary sclerosing cholangitis when compared with other chronic liver diseases. Circulating intercellular adhesion molecule-1 is probably derived from activated lymphocytes rather than from bile ducts because biliary epithelial cells from patients with primary biliary cirrhosis did not release circulating intercellular adhesion molecule-1 when stimulated to express the membrane-bound molecule in vitro. These studies are the first to demonstrate circulating intercellular adhesion molecule-1 in chronic inflammatory diseases that are characterized by strong tissue expression of intercellular adhesion molecule-1 and as such suggest a potential immunoregulatory role for circulating adhesion molecules. The very high levels detected in primary biliary cirrhosis and primary sclerosing cholangitis probably reflect lymphocyte activation, which is further evidence of immune pathogeneses for these diseases.


A high-resolution cytogenetic map of human chromosome 3: localization of 291 new cosmid markers by direct R-banding fluorescence in situ hybridization. We localized 291 new cosmid markers (including 65 RFLPs) on human chromosome 3 by direct R-banding fluorescence in situ hybridization. This system, which is based on fluorescence in situ hybridization combined with replicated prometaphase R-bands, allows the direct visualization of signals on R-banded prometaphases stained with propidium iodide and provides a more rapid and efficient method for genome mapping of cosmid clones. The signals of 291 markers examined here were localized preferentially to R-positive bands throughout chromosome 3. The detailed map positions of 366 clones and the characterization of 142 RFLPs, including the preliminary data reported by Yamakawa et al. (1991, Genomics 9: 536-543; and 11: 565-572), are summarized. This high-resolution cytogenetic map (average distance of 0.58 Mb), in conjunction with a genetic linkage map, can facilitate the analysis of chromosomal and molecular aberrations in genetic diseases and cancers. Furthermore, these mapping data will provide many useful landmarks for the construction of contig maps of chromosome 3.


An interspecific linkage map of mouse chromosome 15 positioned with respect to the centromere. We have used an interspecific backcross between C57BL/6J and Mus spretus to derive a molecular genetic linkage map of chromosome 15 that includes 25 molecular markers and spans 93% of the estimated length of chromosome 15. Using a second interspecific backcross that was analyzed with a centromere-specific marker, we were also able to position our map with respect to the chromosome 15 centromere. This map provides molecular access to many discrete regions on chromosome 15, thus providing a framework for establishing relationships between cloned DNA markers and known mouse mutations and for identifying homologous genes in mice and humans that may be involved in disease.


Isolation of 1001 new markers from human chromosome 11, excluding the region of 11p13-p15.5, and their sublocalization by a new series of radiation-reduced somatic cell hybrids. The determination of the physical map of human chromosome 11 will require more clones than are currently available. We have isolated an additional 1001 new markers in a bacteriophage vector from a somatic cell hybrid cell line that contains most of chromosome 11, except the middle of the short arm. These markers were localized to five different regions, 11p15-pter, 11p12-cen, 11q11-q14, 11q14-q23, and 11q23-qter, by a panel of previously characterized somatic cell hybrids. The region 11q11-14 harbors genes that have been shown to be important in breast cancer, B-cell lymphomas, centrocytic lymphomas, asthma, and multiple endocrine neoplasia, type 1 (MEN1). To determine the positions of the recombinant clones located there, we developed a new series of radiation-reduced somatic cell hybrids. These hybrids, together with those previously characterized, allowed us to map the 11q11-q14 markers into 11 separate segregation groups.


A multipoint genetic linkage map of mouse chromosome 18. We have mapped 13 loci on mouse Chromosome 18 by Southern blot analysis of restriction fragment length polymorphisms among progeny from an interspecific backcross: (C57BL/6J X Mus spretus) X M. spretus. Complete haplotype analysis of 136 of these progeny was used to establish gene order and estimate genetic distances between loci. The gene order (from centromere to telomere) and recombination distances (in centimorgans) were as follows: PGK-1rs5-4.3-Tpi-10-11.8-(Egr-1, Hmg17-rs9)-2.1-Fgfa-2.2-Grl-1-10.1-(Cdx-1, Csfmr, Pdgfrb, Pdea, Rps14)-2.1-Adrb-2-22.9-Mbp. Pgk-1rs5, Tpi-10, Hmg17-rs9, and Rps14 had not been previously mapped in the mouse; Egr-1 had only been syntenically assigned to mouse Chr 18. Nine of the loci, spanning 18 cM, have homologs on the distal long arm of human Chr5--a region rich in genes encoding growth factors and receptors. An additional previously unmapped gene, Drd-1, predicted to be on mouse Chr 18 based on its human chromosomal location, was mapped to the middle region of mouse Chr 13.


DLX2 (TES1), a homeobox gene of the Distal-less family, assigned to conserved regions on human and mouse chromosomes 2. Dlx-2 (also called Tes-1), a mammalian member of the Distal-less family of homeobox genes, is expressed during murine fetal development in spatially restricted domains of the forebrain. Searching for a candidate neurological mutation that might involve this gene, we have assigned the human and mouse loci to regions of conserved synteny on human chromosome 2, region cen--q33, and mouse chromosome 2 by Southern analysis of somatic cell hybrid lines. An EcoRI dimorphism, discovered in common inbred laboratory strains, was used for recombinant inbred strain mapping. The results place Dlx-2/Tes-1 near the Hox-4 cluster on mouse chromosome 2.


Polymorphisms and deduced amino acid substitutions in the coding sequence of the ryanodine receptor (RYR1) gene in individuals with malignant hyperthermia. Twenty-one polymorphic sequence variants of the RYR1 gene, including 13 restriction fragment length polymorphisms (RFLPs), were identified by sequence analysis of human ryanodine receptor (RYR1) cDNAs from three individuals predisposed to malignant hyperthermia (MH). All RFLPs were detectable in PCR-amplified products, and their segregation was consistent with our initial finding of linkage to MH in the nine families previously informative for one or more intragenic markers (MacLennan et al., 1990, Nature 343:559-561). Four amino acid substitutions were identified in the study: Arg for Gly248, Cys for Arg470, Leu for Pro1785, and Cys for Gly2059. Of 45 families tested, a single family presented the Arg for Gly248 substitution where it segregated with malignant hyperthermia, making it a candidate mutation for predisposition to MH in man. The other three polymorphic substitutions failed to segregate with malignant hyperthermia in those families in which they occurred, implying that they represent polymorphisms with little or no effect on the function of the RYR1 gene.


Isolation and mapping of 88 new RFLP markers on human chromosome 8. To obtain new RFLP markers for construction of a high-resolution map of human chromosome 8, a cosmid library was constructed from a somatic hybrid cell that contained chromosome 8 as the only human component in mouse genomic background. Eighty-eight new RFLP markers were isolated and characterized, and 71 of them were sublocalized to chromosomal bands by fluorescent in situ hybridization (FISH). Of these, 36 were localized to the short arm, 34 to the long arm, and 1 to the centromeric region. Five markers defined VNTR loci. This work represents the first extensive isolation and physical mapping of RFLP markers on human chromosome 8. These new markers will serve as useful resources for linkage mapping of loci for inherited diseases and for efforts to identify a putative tumor suppressor gene(s) on chromosome 8.


A molecular genetic linkage map of mouse chromosome 18 reveals extensive linkage conservation with human chromosomes 5 and 18. An interspecific backcross between C57BL/6J and Mus spretus was used to generate a molecular genetic linkage map of mouse chromosome 18 that includes 23 molecular markers and spans approximately 86% of the estimated length of the chromosome. The Apc, Camk2a, D18Fcr1, D18Fcr2, D18Leh1, D18Leh2, Dcc, Emb-rs3, Fgfa, Fim-2/Csfmr, Gnal, Grl-1, Grp, Hk-1rs1, Ii, Kns, Lmnb, Mbp, Mcc, Mtv-38, Palb, Pdgfrb, and Tpl-2 genes were mapped relative to each other in one interspecific backcross. A second interspecific backcross and a centromere-specific DNA satellite probe were used to determine the distance of the most proximal chromosome 18 marker to the centromere. The interspecific map extends the known regions of linkage homology between mouse chromosome 18 and human chromosomes 5 and 18 and identifies a new homology segment with human chromosome 10p. It also provides molecular access to many regions of mouse chromosome 18 for the first time.


Extension of the physical map in the region of the mouse X chromosome homologous to human Xq28 and identification of an exception to conserved linkage. We have extended our pulsed-field gel map of the region of the mouse X chromosome homologous to human Xq28 to include the loci Gdx (DXS254Eh), P3 (DXS253Eh), G6pd, Cf-8, and F8a. Gdx, P3, and G6pd are demonstrated to be physically linked to the X-linked visual pigment locus (Rsvp) within a maximal distance of 340 kb, while G6pd and Cf-8 are approximately 900 kb apart. These studies favor a gene order of cen-Rsvp-Gdx-P3-G6pd-(Cf-8)-tel and extend the physical map of this region to 5 million bp. In conjunction with previous physical mapping studies in both mouse and human, the results suggest conserved linkage for loci in this region of the mouse X chromosome and human Xq28. However, employing pulsed-field gel electrophoresis and genetic pedigree analysis of interspecific backcross progeny, we have found close linkage of a clone encoding a mouse homolog for human factor VIII-associated gene A (F8A) to DXPas8, thus revealing the first exception to conserved gene order between murine and human loci in the region.


Identification of 57 conventional RFLP and 6 VNTR systems with 32 DNA clones on chromosome 11p15. Fifty-four clones containing human inserts were selected from a cosmid library constructed from a somatic cell hybrid containing chromosome 11p15.3-p15.5 as its only human complement. In 32 of these clones, 63 polymorphic systems were identified with a panel of restriction enzymes: 57 conventional RFLP systems and 6 highly polymorphic VNTR systems. Although we examined the cosmid with only seven enzymes, 18 clones (including 6 VNTRs) were polymorphic with three or more enzymes. The results suggested that DNA sequences on the peritelomeric region of chromosome 11p tend to be highly variable. Because these markers are highly informative, they will be excellent resources for investigations of hereditary diseases and tumor suppressor genes in this region of chromosome 11.


Regional localization of three convertases, PC1 (Nec-1), PC2 (Nec-2), and furin (Fur), on mouse chromosomes. The genes for three convertases, PC1 (Nec-1), PC2 (Nec-2), and furin (Fur), have been regionally localized on chromosomes 13, 2, and 7, respectively, by interspecific backcross analysis. These results refine previous localizations by in situ hybridization as well as confirm and extend known regions of homology between mouse and human chromosomes.


Assignment of the gene coding for hepatocyte growth factor-like protein to mouse chromosome 9. The gene coding for hepatocyte growth factor-like protein has been localized to mouse chromosome 9 at a locus (Hgfl) distal to the Trf locus. The likely gene order in this region is centromere-Trf-Gnai-2-Hgfl-Cck. The region surrounding the Hgfl locus shows homology of syntenty to band p21 on human chromosome 3.


Blood glucose response to stress hormone exposure in healthy man and insulin dependent diabetic patients: prediction by computer modeling. To establish a qualitative and quantitative model of blood glucose response to stress hormone exposure, healthy subjects (HS) on and off somatostatin (250 micrograms/h) as well as insulin dependent diabetic patients were infused with either epinephrine (E), glucagon (G), cortisol (F), growth hormone (GH) or with a cocktail of these hormones raising plasma stress hormones to values seen in severe diabetic ketoacidosis. The developed input/output model consists of two submodels interconnected in series plus two additional submodels for correction of gains describing both sensitivity of tissue response and utilisation as well as provision of glucose. It was shown and confirmed experimentally that blood glucose response to stress hormones was essentially nonlinear. Furthermore, the mathematical models for healthy subjects and for insulin dependent diabetic patients proved to be of the same structure and differed only in the values of some typical parameters. The model raises the possibility to describe and in part to predict blood glucose response to stress hormone exposure in healthy man and insulin dependent diabetic patients.


Role of T-cell derived cytokines in the downregulation of immune responses in parasitic and retroviral infection. Parasitic infection is frequently accompanied by a downregulation in host cell-mediated immunity. Recent studies suggest that this modulation of helper T cells and effector cell function can at least in part be attributed to the action of a set of inhibitory cytokines produced by T lymphocytes as well as by a number of other cell types. The best characterized of these inhibitory lymphokines are IL-4, IL-10 and TGF-beta. Interestingly, both IL-4 and IL-10 are produced by the Th2 but not the Th1 subset of CD4+ helper cells. The former subset dominates in many situations of chronic or exacerbated parasitic infection and is thought to suppress Th1 function as a consequence of the cross-regulatory activity of these two cytokines. The latter hypothesis is supported by recent experiments demonstrating that mAb-mediated neutralization of IL-10 reverses suppressed IFN-gamma responses and/or disease susceptibility in mice with parasitic infections. In vivo neutralization of TGF-beta has also been reported to increase host resistance to parasite challenge. In addition to suppressing T-cell differentiation, function or proliferation, IL-4, IL-10 and TGF-beta each inhibit the ability of IFN-gamma to activate macrophages for killing of both intracellular and extracellular parasites. Moreover, the three cytokines are able to synergize with each other in downregulating these parasiticidal effects. Interestingly, each of the cytokines inhibits the production of reactive nitrogen oxides, an effector mechanism previously demonstrated to play a major role in parasite killing by activated macrophages. In the case of IL-10, this suppression of nitrogen oxide production appears to result from an inhibition of TNF-alpha synthesis leading to defective macrophage stimulation. While distant from parasites in their biology and phylogeny, some retroviruses also appear to induce an over-production in downregulatory cytokines which is closely associated with the onset of immunodeficiency. Thus, in an animal model involving infection of mice with LP-BM5 MuLV and in human HIV infection, Th2 (IL-10 and/or IL-4) cytokine synthesis is increased while Th1 (IFN-gamma and/or IL-2) cytokine production is suppressed. These observations suggest that cytokine-mediated cross-regulation may play a role in the pathogenesis of acquired immune deficiency disease, contributing both to the progression of retroviral infection and the increase in susceptibility to opportunistic infections and malignancy. Observations of similar cytokine cross-regulatory activities in organisms as diverse as helminths, protozoa and retroviruses predict that comparable mechanisms may operate in a wide variety of infectious diseases.


Effect of organophosphate pesticides on glutaminase synthetase activity in rat brain. Of the organophosphate pesticides studied, dichlorvos inhibited significantly both, phosphate-activated glutaminase and alpha-keto acid-activated glutaminase, while monocorotophos inhibited moderately alpha-keto acid activated glutaminase in rat brain. Phosphamidon inhibited glutamine synthetase activity negligibly.


Occurrence of colonization factor antigens I & II in enterotoxigenic Escherichia coli associated diarrhoea in Iran & correlation with severity of disease. The occurrence of colonization factor antigens I and II (CFA/I and II) and type 1 somatic pili was investigated in 197 enterotoxigenic Esch. coli (ETEC) isolated from 197 patients of diarrhoea (aged under 3 yr) during February 1985 to March 1986 in Tehran, Iran. Among ETEC strains, 154 strains were heat-stable enterotoxin (ST) producers, 27 strains were heat-labile enterotoxin (LT) producers, and 16 strains produced both toxins. Sixty five (33%) strains showed mannose-resistant haemagglutination (MRHA) of human and/or bovine erythrocytes; of these, 51 (86%) strains were positive for CFA/I and II. Seventy one (36%) strains also exhibited type 1 somatic pili. CFA/I was found in 4 (15%) LT producing, 24 (16%) ST producing, and 2 (13%) LT/ST producing strains. In contrast, CFA/II was only found in ST producing strains (17 strains) and those producing both toxins (4 strains). Patients having CFAs-positive ETEC strains had a significantly (P less than 0.001) higher number of stool evacuation per day and a longer duration of diarrhoea than those having CFAs-negative strains. Fifty nine patients had mixed infections of ETEC strains and other enteropathogens. CFA/I or II (CFAs)-positive and CFAs-negative ETEC strains were found in 17 and 42 patients with mixed infections respectively. The mean number of stool evacuations per day was much higher in patients with ETEC and rotavirus than those with only ETEC infection (P less than 0.001). However, severity of the disease was not affected by the presence or absence of CFA/I or II in ETEC strains found in these patients.


Undescended testis: evaluation by magnetic resonance imaging. We describe our experience of prospective magnetic resonance imaging (MRI) study in patients of undescended testis, with a 1.5 T equipment using body coil. There were thirty two patients, aged 1.5 to 14 years with a mean age of nine years. Surgical follow up was obtained for thirty one patients. We were able to indicate the position of 26 testes in 22 patients and absence of five testes in three patients. MRI was falsely positive and negative for five and two testes, respectively. Testicular tissue at ectopic site was identified by presence of characteristic signal intensity pattern, mediastinum testis and its location along empty spermatic canal in cases of inguinal testis either singly or in combination. MRI was able to detect atrophic changes in four testes, confirmed on surgery. The study concludes that MR imaging is useful in the localization and tissue characterization of a non palpable testis. However, it is not sensitive enough for complete exclusion of the diagnosis of an undescended testis. Thus a surgical or laproscopic exploration may be needed further in selective cases for the management of patient.


Myocardial anti-ischemic characteristics of a novel class of beta-adrenoceptor blockers. The effect of different beta-adrenoceptor blockers on free radical-mediated cardiac membrane lipid peroxidation (CMLP) was compared to their beta-blocking potency (pA2). CMLP was determined by the measurement of malondialdehyde (MDA) formation in rat cardiac membrane homogenates exposed to a free radical generating system (FeCL3/ADP/DHF) in the presence or absence of the beta-adrenoceptor blockers (1-1,000 microM). beta-adrenoceptor blocking potency (pA2) was determined using guinea pig right atria stimulated with isoproterenol. The catechol containing beta-blocker (DCC-10255) was shown to be a potent inhibitor of CMLP via an iron-dependent mechanism. On the other hand, the non-catechol beta-adrenoceptor blocker, timolol, was shown to be a weak inhibitor of CMLP. Furthermore, dl- and d-propranolol were active and equipotent though less potent than DCC-10255 in inhibiting CMLP. The myocardial cytoprotective efficacy for catechol and non-catechol beta-adrenoceptor blockers was evaluated in an isolated rat myocyte model. It was demonstrated that catechol-containing agents with either strong or weak beta-adrenoceptor blockade possess a relatively potent in vitro antioxidant and myocardial cytoprotective efficacy against free radical mediated CMLP and myocyte injury, respectively. It is concluded that the inhibition of CMLP by beta-adrenoceptor blockers is independent of their beta-adrenoceptor blockade.


Comparative review of third-generation progestins. Desogestrel, gestodene, and norgestimate represent a new generation of progestins designed for use in oral contraceptives. A high degree of efficacy has been retained in these progestins, and the adverse metabolic impact exhibited by older progestins has been reduced considerably. The clinical profile of each progestin, as it is marketed in Europe in combination with ethinyl estradiol, is reviewed. Direct comparisons are made whenever applicable. The major advantages of these formulations over the older combined oral contraceptives are that they have less effect on lipid metabolism and on carbohydrate metabolism, and they are less androgenic. The clinical implications of these findings are discussed.


Women, lipoproteins, and cardiovascular disease risk. Women, like men, are susceptible to coronary atherosclerosis. Like men, more women die of heart disease than all forms of cancer combined. Coronary atherosclerosis is the result of the interplay of a number of factors, the most important of which are abnormal levels of circulating lipoproteins. As more has become known about the mechanisms by which abnormal levels of circulating lipoproteins promote atherosclerosis, certain risk factors have emerged as concerns for women, including: (1) diabetes mellitus as a risk factor, perhaps through its more profound effects on circulating lipoproteins; (2) serum triglyceride levels, and (3) changes in high-density lipoprotein cholesterol. The widespread use of exogenous hormones in women as both oral contraceptives and postmenopausal hormone replacement may also play a role in developing atherosclerosis. In general, estrogen affects circulating lipoprotein levels favorably, whereas progestins have the opposite effect. The effects of estrogen/progestin combinations in either oral contraceptives or postmenopausal hormone replacement will depend on the relative dose and potency of each of these constituents. Epidemiologic studies indicate that the use of oral contraceptives has no profound effect on the long-term risk of heart disease, whereas unopposed estrogen (without progestin) in postmenopausal hormone replacement therapy may lower that risk considerably. Recent U.S. and European guidelines for the detection, evaluation, and treatment of hypercholesterolemia in adults make it imperative that obstetrician-gynecologists, in their dual role as primary-care physicians and prescribers of exogenous hormones, be aware of and informed about the relationship between circulating lipids and lipoproteins and coronary heart disease in women.


Diabetes mellitus, hypertriglyceridemia, and heart disease risk in women. Coronary heart disease is the most common cause of death in men and women in developed countries. Three primary risk factors--high serum cholesterol concentration, hypertension, and cigarette smoking--are known to increase the risk in both men and women more or less equally, although the latter two risk factors are a somewhat greater risk to men. This paper reviews two additional risk factors whose impact may be greater in women: diabetes and hypertriglyceridemia. Understanding how diabetes and hypertriglyceridemia act differently in women may explain some of the sex differences in the risk of heart disease.


Mechanism of action/effects of androgens on lipid metabolism. The strongest predictors of cardiovascular disease in women have been shown to be diabetes, high blood pressure, cigarette smoking, and, to a lesser degree, hypertriglyceridemia. The difference in risk between men and premenopausal women has been explained by the following widely held hypothesis: androgens lower plasma concentrations of high-density lipoprotein (HDL), particularly the HDL-2 subfraction, and increase plasma concentrations of low-density lipoprotein (LDL). In contrast, estrogens have the opposite effect, raising plasma concentrations of HDL, particularly HDL-2, and lowering plasma concentrations of LDL. After the menopause, it is believed that the protective effect of estrogens in women is lost and the incidence of heart disease rises to equal that in men. This paper provides a brief review of the effect of endogenous and exogenous androgens on lipoprotein metabolism in men and women, and considers the relevance of these findings to the choice of progestogens used in oral contraceptive preparations.


Steroids and lipid metabolism: mechanism of action. The effects of steroids on hepatic metabolism depend on the preexisting metabolic status of the individual and the structure, dosage, and, to a certain extent, the route of administration of the steroid compound. Oral administration of synthetic estrogen causes an increase in hepatic production of very-low-density lipoprotein (causing in turn an increase in triglyceride levels), an increase in coagulation factors, and an increase in high-density-lipoprotein cholesterol, which is related to a decrease in hepatic lipase activity. Parenteral administration of estrogen does not appear to cause any such modification. The effects of progestogen on hepatic metabolism are dependent on the androgenic activity of the specific progestogen administered. A progestogen with greater androgenic activity causes a decrease in high-density-lipoprotein cholesterol, which is related to an increase in hepatic lipase activity. This article reviews the effects of steroids (both estrogens and progestogens) on the mechanisms of action in lipid metabolism.


Effects of hormone replacement therapy.

Effects of hormone replacement therapy. According to the World Health Organization, the global life expectancy at birth was 55 years in 1974, will be 63 years in 2000, and will be close to 70 years in the year 2025. By the year 2025, approximately 20% of the world's population will be more than 60 years old, and the problems associated with hormonal changes become significant with more than one third of a woman's expected life concentrated in the postmenopausal years. Much evidence suggests that the use of postmenopausal estrogen substantially reduces the morbidity and mortality of coronary heart disease, and to date, low-dose estrogen therapy is the single most effective method for prevention of osteoporosis. However, only hysterectomized women can use estrogen alone because of the risk of endometrial cancer associated with unopposed estrogen therapy. Thus, women with an intact uterus must use estrogen in combination with progestogen to avoid this risk, but the different progestogens may variously modify the beneficial effects of estrogen on lipoproteins, and, ultimately, coronary heart disease. Although hormone replacement therapy during menopause soon will be one of the major areas of preventive medicine, the effects of estrogen in combination with different progestins on coronary heart disease, blood pressure, blood coagulation, bone density, and the central nervous system have not been investigated adequately. This paper reviews the current knowledge of known or suspected effects.


Augmentation by bispecific F(ab')2 reactive with P-glycoprotein and CD3 of cytotoxicity of human effector cells on P-glycoprotein positive human renal cancer cells. A bispecific F(ba')2 was constructed that was composed of two Fab fragments, one derived from anti-CD3 monoclonal antibody (mAb) (OKT3) and the other from anti P-glycoprotein mAb (MRK 16). This bispecific F(ab')2 enhanced the binding and cytotoxicity of human peripheral blood mononuclear cells (PBMCs) on P-glycoprotein-positive human kidney cancer cells (ADMHK/E). It had no effect on the cytotoxicity of PBMCs on P-glycoprotein-negative HK/E cells [long-term cultured HK/E (LCHK/E)]. Control F(ab')2 composed of OKT3 or MRK16 alone did not influence the cytotoxicity of PBMCs on ADMHK/E cells. These findings suggest that the MRK16-OKT3 bispecific F(ab')2 may be therapeutically beneficial in treatment of human multidrug-resistant cancers.


Intravenous recombinant secretory leukoprotease inhibitor augments antineutrophil elastase defense. Secretory leukoprotease inhibitor (SLPI), a 12-kDa serine antiprotease, normally protects the upper airway epithelial surface from attack by neutrophil elastase (NE). In the context that a variety of inflammatory lung diseases are characterized by large neutrophil burdens with resultant high levels of NE in the lung, recombinant SLPI (rSLPI), a molecule identical to natural SLPI, may be an effective means to augment the anti-NE protective screen of the lung. To determine whether intravenous rSLPI will augment respiratory tract and epithelial surface levels of SLPI and anti-NE capacity, rSLPI was administered intravenously to sheep and SLPI levels were quantified in plasma, lung lymph (as a measure of lung interstitial levels), lung epithelial lining fluid (ELF), and urine. rSLPI (1 g) was administered over 10 min, and after 30 min plasma levels of SLPI were 8 microM and decreased with a half-life of 1.8 h. Lymph SLPI levels paralleled the plasma levels: 4 h after infusion the lymph-to-plasma ratio was 0.8. ELF SLPI levels paralleled the lymph levels: 4 h after infusion the ELF-to-lymph ratio was 0.3. Western analysis demonstrated intact SLPI in lymph and ELF, and functional analysis showed increases in lymph and ELF anti-NE capacity that paralleled the levels of SLPI. As might be expected from a protein with a molecular mass of 12 kDa, urine excretion was high, with 20% of the SLPI excreted over 5 h. However, if the rate of infusion was slowed, SLPI excretion decreased significantly, with a 3-h infusion associated with 9% excretion and a 12-h infusion associated with less than 0.2% excretion.(ABSTRACT TRUNCATED AT 250 WORDS)


Characterization of a simian virus 40-transformed human podocyte cell line producing type IV collagen and exhibiting polarized response to atrial natriuretic peptide. Biology of glomerular visceral epithelial cells ("podocytes") and their role in inflammatory process remain obscure, partly because of the lack of well-differentiated podocyte cultures. We have established a human cell line by transfecting with a replication-defective SV40 plasmid (pSVHB1), a primary culture of podocytes derived from an enriched preparation of unencapsulated glomeruli free of tubule and Bowman's capsule contaminants. Podocyte specificity of the primary culture was assessed by a dual immunomorphological and functional approach. The resulting cell line (HGVEC.SV1) was cloned and the clonal cells were adapted to hormonally defined medium supplemented with only 2% newborn bovine serum. Clone A4 has been exhibiting over 35 passages, a combination of markers unique to podocytes, including expression of vimentin, podocalyxin, ectoenzymes (CALLA antigen and mRNA), heparan-sulfate proteoglycans (molecular mass of core protein = 75 kDa), and production of type IV collagen (alpha 1 and alpha 5 chains) established by immunoprecipitation and Northern blot analysis. Cytokeratin was detected in rare cellular foci and the search of Von Willebrand factor was negative. This clonal cell line has been used to demonstrate: (1) that human podocytes are highly sensitive to atrial natriuretic peptide (ANP) which induced a dose-dependent increase in cGMP production (x20 at 0.5 microM ANP), and (2) that secretion of ANP-stimulated cGMP is dramatically polarized as 93% of extracellular cGMP were released in the apical medium when filter-grown HGVEC. SV1A4 cells were stimulated at their basal pole.


Restoration of proliferating cell nuclear antigen (PCNA) complex formation in xeroderma pigmentosum group A cells following cis-diamminedichloroplatinum (II)-treatment by cell fusion with normal cells. We examined the role of the factor deficient in xeroderma pigmentosum group A (XP-A) cells in the formation of proliferating cell nuclear antigen (PCNA) complex with DNA in the DNA repair process in human fibroblasts following cis-diamminedichloroplatinum (CDDP)-treatment. Immunofluorescence staining after methanol fixation was used to detect the PCNA complex formation. When quiescent normal cells were PCNA-stained at 3 h after 100 microM CDDP treatment for 1 h, almost all nuclei of the cells showed a punctuated staining pattern. On the other hand, nuclei of XP-A cells were not stained. These results were the same with the findings following 10J/m2 of ultraviolet light (UV)-irradiation. The quantitative analysis of the PCNA immunofluorescence intensity of normal cells revealed that the mean intensity was increased by 4.8 times by the CDDP-treatment and 6.1 times by the UV-irradiation, compared with that of untreated cells. The intensities among nuclei ranged widely in both treatments. In contrast, the mean intensity was not increased in XP-A cells by the same treatments. However, when XP-A cells were fused with normal cells with polyethylene glycol (PEG) treatment, the nuclei of the XP-A cells showed positive PCNA-staining following CDDP-treatment or UV-irradiation in almost all cases. These results suggest that the PCNA complex formation may play a role in the DNA repair process after the step where the factor deficient in XP-A cells is involved following CDDP-treatment as well as following UV-irradiation.


Conditions for assembly of tubulin-based structures in unfertilized sea urchin eggs. Spirals, monasters and cytasters. Cytasters were induced in the unfertilized eggs of the sea urchin Lytechinus pictus by two different methods: (1) treatment with hexylene glycol or taxol, which are known to lower the tubulin critical concentration in vitro, but do not activate the nuclear cycle, and (2) by raising the cytoplasmic pH from that of unfertilized cytoplasm to that of fertilized, which activated the nuclear cycle and initiated tubulin polymerization. In the unactivated eggs, with increasing concentrations of the inducing agents, temperature and duration of treatment, microtubule structures that formed showed a progression from loose microtubule networks and spiral arrays to tightly organized cytasters. In eggs incubated in sea water containing 2.5 or 10 mM ammonium acetate titrated with HCl or NaOH in steps from pH 5.0 to pH 9.0, the nuclear cycle and tubulin polymerization were initiated at about pH 7.0. The degree of development attainable after three hours was dependent on the pH, with spirals forming at the threshold level of pH 7.0, monasters at pH 7.5, and at pH 8.5 cells formed cytasters, multipolar spindles and even completed multipolar divisions. In unactivated eggs, tubulin was made available by lowering of the tubulin critical concentration; in activated eggs it was made available from some previously unavailable store. The evidence suggests that the amount of assembly-competent tubulin available, regardless of how it is made available, determines the type of structure that is formed, with microtubule bundles and spirals at the lower concentrations and functional cytasters at the upper. We also describe some details of cytaster formation, including the role of microtubule interactions and the movement of dense granules to the aster centers in hexylene glycol-treated eggs.


Radioimmunoassay for the synthetic ergoline derivative cabergoline in biological fluids. An antiserum against cabergoline, a powerful dopamine-agonist under clinical trials for the treatment of Parkinson's disease and hyperprolactinemia, has been raised in rabbits by immunization with an immunogen produced by conjugation of cabergoline to bovine serum albumin. The antiserum was able to bind a derivative of cabergoline labelled with tritium and was able to distinguish the drug molecule from some of its close related compounds and from other agents that could be simultaneously present in plasma from patients undergoing treatment with cabergoline. The antiserum and the tritium labelled hapten were used to develop a radioimmunoassay for cabergoline determination in human plasma and urine. A linear relationship between cabergoline added and % radioactivity bound was found in the range 1.9-500 pg/tube. The addition in the assay of 200 microliters human plasma or 25 microliters urine did not affect the specific and the non-specific binding of the radiolabelled hapten so enabling us to obtain a final sensitivity of about 12 pg/ml plasma and 120 pg/ml urine. The assay was validated in terms of reproducibility, precision and accuracy over the whole range of concentrations tested both in plasma and urine. The plasma concentrations at the steady state in a patient with Parkinson's disease who had received the drug at single oral daily doses of 3, 5 and 7 mg were determined using the assay.


Immunoglobulins in milk from cows immunized with oral strains of Actinomyces, Prevotella, Porphyromonas, and Fusobacterium. Immunization of pregnant cows with bacteria leads to the presence of high concentrations of specific antibodies in colostrum and milk. A total of 14 cows was immunized with single strains of heat-killed oral bacteria or pools of strains of Actinomyces, Porphyromonas, Prevotella, and Fusobacterium. Two cows were treated with adjuvant alone. The mean percentages of IgG1, IgG2, IgM, and IgA in all of the milks were 83.8, 3.8, 9.3, and 3.1, respectively. ELISA and whole cell agglutination assays demonstrated high titers in the milks from the cows immunized with either individual strains or the bacterial pools. The highest titers determined by ELISA belonged to the IgG1 isotype and in several milks were 64-fold greater than titers in milk from cows treated with adjuvant alone. The concentrations of all antibodies and the titers determined by ELISA and whole cell agglutination assays markedly decreased from the first to the sixth milkings. The functional specificity of the antibodies was demonstrated by agglutination tests against a wide range of bacteria including members of Actinomyces, Fusobacterium, Porphyromonas, Prevotella, Streptococcus, Eubacterium, Propionibacterium, Peptostreptococcus, Bacteroides, Actinobacillus, Haemophilus, Capnocytophaga, and Wolinella. Minimal cross-reactions with bacteria in other genera were observed with all of the milks. High-titer milk preparations have been obtained from immunized cows, and the capacity of the bovine antibodies to agglutinate target bacteria indicates their potential usefulness in oral passive immunization studies.


Common purslane: a source of omega-3 fatty acids and antioxidants. omega-3 fatty acids, alpha-tocopherol, ascorbic acid, beta-carotene and glutathione determined in leaves of purslane (Portulaca oleracea), grown in both a controlled growth chamber and in the wild, were compared in composition to spinach. Leaves from both samples of purslane contained higher amounts of alpha-linolenic acid (18:3w3) than did leaves of spinach. Chamber-grown purslane contained the highest amount of 18:3w3. Samples from the two kinds of purslane contained higher leaves of alpha-tocopherol, ascorbic acid and glutathione than did spinach. Chamber-grown purslane was richer in all three and the amount of alpha-tocopherol was seven times higher than that found in spinach, whereas spinach was slightly higher in beta-carotene. One hundred grams of fresh purslane leaves (one serving) contain about 300-400 mg of 18:3w3; 12.2 mg of alpha-tocopherol; 26.6 mg of ascorbic acid; 1.9 mg of beta-carotene; and 14.8 mg of glutathione. We confirm that purslane is a nutritious food rich in omega-3 fatty acids and antioxidants.


Long-term haemodynamic effects of a 4-week regimen of nipradilol, a new beta-blocker with nitrovasodilating properties, in patients with portal hypertension due to cirrhosis. A comparative study with propranolol. To study the long-term effects of pharmacological combination therapy, a comparison was made of the haemodynamic changes in patients with cirrhosis and portal hypertension following a 4-week treatment of propranolol or nipradilol, a new nonselective beta-blocker with nitrovasodilating effect. Nipradilol (12 mg/dag, n = 12) significantly diminished wedged hepatic venous pressure (WHVP, 25 +/- 16%), the hepatic venous pressure gradient (HVPG, 20 +/- 12%), and estimated hepatic blood flow (EHBF, 18 +/- 16%). Propranolol (30 mg/day, n = 11) also caused a significant reduction in WHVP (22 +/- 21%) and HVPG (24 +/- 21%), but not in EHBF. The percentage of portal pressure reduction and the frequency of nonresponders did not differ between the nipradilol and propranolol groups. Both agents reduced heart rate by approx. 20%. Nipradilol, however, did not cause a significant reduction in cardiac index (CI) versus a 14% reduction by propranolol. Pulmonary capillary wedge pressure and central venous pressure, an index of preload, were decreased slightly in the nipradilol group. When nonresponders were excluded, there was a significant correlation of the percentage of reduction between WHVP and CI or systemic vascular resistance, in the nipradilol group. These results indicate that nipradilol may have potent hypotensive effects on portal hypertension, similar but not superior to propranolol. Nipradilol, at the dosage used in the present study, did not appear to exert a nitrovasodilating effect to enhance the portal pressure reduction induced by beta-blocking action.


Detection of proliferating cell nuclear antigen in diagnostic histopathology. We describe the effects of tissue preservation, fixation time, and hydrolytic treatment on the detection of proliferating cell nuclear antigen (PCNA) by immunoperoxidase staining with three commercial anti-PCNA antibodies (19A2, 19F4, PC10). Our goal was to provide guidelines for PCNA immunohistochemistry in formalin-fixed, paraffin-embedded specimens. In proliferative cell compartments, nuclear staining was achieved with all three antibodies. In some cases PCNA was also expressed in non-proliferative, histologically normal tissues associated with tumors or other lesions elsewhere. In most autopsy specimens PNCA immunoreactivity was markedly diminished as compared with similar surgical specimens. Incubation overnight with primary antibody at 4 degrees C enhanced PCNA immunoreactivity over incubation at 42 degrees C for 45 min. Pre-treatment with 2 N HCl did not increase staining. Staining with the PC10 antibody was much better preserved than staining with the antibodies 19A2 and 19F4 after prolonged formalin fixation of surgical specimens and in tissues obtained at autopsy. With all three antibodies, however, PCNA immunoreactivity was well preserved during formalin fixation for 8-24 hr and during fixation delays for 8 hr at room temperature. This indicates that PCNA is stable under conditions routinely encountered in diagnostic surgical pathology and facilitates its potential use as a diagnostic proliferation marker.


Generation and characterization of IL-2-activated veto cells. The regulation of in vivo cytolytic response is important in a model of murine graft-vs-host disease induced by the injection of parental splenocytes into unirradiated B6D2F1 recipients. Injection of C57BL/6J spleen cells into B6D2F1 recipients results in an acute form of graft-vs-host disease that is characterized by the presence of CTL and suppressor cells, runting, and occasionally death. In contrast, injection of DBA/2J spleen cells into B6D2F1 recipients results in a chronic form of graft-vs-host disease that is characterized by the lack of in vivo CTL and hyperproduction of Ig and autoantibodies that results in an SLE-like syndrome. One reason for the lack of donor antirecipient CTL after injection of DBA/2J donor cells is that B6D2F1 recipient cells functionally inactivate the donor DBA/2J CTL precursor cells by expressing veto activity. These B6D2F1 veto cells are radiosensitive, inhibited by anti-CD8 antibodies, found primarily in lymph nodes, and were further characterized by testing the response of these inhibitory cells to lymphokines. These studies indicate that IL-2 can potentiate the activity of the veto cells induced in vivo and veto cells with a similar phenotype can be generated by in vitro incubation of naive lymph node cells with IL-2. These cells have been designated as IL-2-activated veto cells or LAV cells. IL-2 did not increase inhibitory activity by increasing the number of CD8+ cells or the number of CD8 molecules on the LAV cell surface but by altering the activation state of the LAV cell. The inhibitory capabilities of antibodies binding various cell surface molecules indicated that CD2 and intercellular adhesion molecule-1 molecules in addition to CD8 molecules played a role in the function of LAV cells.


Fetal skin: a site of dendritic epidermal T cell development. Thy-1 Ag and CD3-associated TCR-gamma (V gamma 3)/delta (V delta 1) are coexpressed on virtually all dendritic epidermal T cells (DETC) in the adult mouse. In contrast, day 16 fetal mouse skin contains small numbers of CD45+/Thy-1+/CD3- but no CD3+ cells. To see whether the CD45+/Thy-1+/CD3- fetal skin cells can qualify as DETC precursors, we transplanted day 16 fetal skin of C57BL/6 (Thy-1.2) mice onto adult B6Pl-Thy-1a (Thy-1.1) animals. At certain time points after transplantation, grafts were analyzed for the presence of Thy-1 and CD3/TCR Ag. Examination of the grafts, 4 days after transplantation, revealed the presence of few donor-type Thy-1.2+/CD3- and some Thy-1.2+/CD3+ epidermal cells of either round or dendritic configuration. At 10 weeks after transplantation, essentially all CD45+/Thy-1.2+ epidermal cells were anti-TCR V gamma 3 and anti-CD3-reactive, displayed a uniformly dendritic configuration, and, thus, represent DETC. Our assumption that CD45+/Thy-1+/CD3- cells are the only lymphocytes within day 16 fetal skin gained additional support by the observations: 1) that unfractionated as well as anti-CD45 gated single cell suspensions prepared from day 16 fetal skin were consistently devoid of anti-CD3 epsilon and anti-TCR V gamma 3-reactive cells; and 2) that stimulation of these cell suspensions with either Con A plus IL-2 or IL-2 alone regularly resulted in the outgrowth of CD45+/Thy-1+/CD3-/TCR V gamma 3- cells, but never in the appearance of CD45+/Thy-1+/CD3+/TCR V gamma 3+ cells. Our additional finding that Con A plus IL-2- or IL-2-stimulated day 16 fetal skin cells and cell lines derived therefrom contain transcripts of some (CD3 gamma, TCR C beta, TCR C gamma 1, TCR C gamma 4) but not of other (CD3 delta, CD3 epsilon, TCR C alpha, TCR C delta) genes encoding the CD3/TCR complex suggests that Thy-1+/CD3- fetal murine skin cells are of T cell lineage. We therefore propose that the fetal skin microenvironment can provide the stimuli promoting growth and maturation of CD3/TCR V gamma 3/V delta 1-expressing DETC from their CD3- precursors.


Impaired calcium mobilization in CD4+ and CD8+ T cells in a retrovirus-induced immunodeficiency syndrome, murine AIDS. After infection with LP-BM5 murine leukemia viruses, susceptible strains of mice develop a severe and progressive immunodeficiency disease, termed murine AIDS (MAIDS), features of which include markedly impaired T cell response to mitogens or specific Ag stimulation and decreased production of IL-2. Since an elevation of intracellular calcium concentration resulting from binding of Ag to the TCR is associated with IL-2 production, T cells from mice either uninfected or infected with LP-BM5 murine leukemia viruses were examined by a calcium mobilization assay. Both CD4+ and CD8+ T cells from infected mice manifested impaired calcium mobilization responses upon in vitro stimulation with anti-CD3 mAb or Con A. The abnormalities appeared early after virus inoculation and showed no difference in time course between subsets of T cells. Frequencies of prestimulation calcium-positive cells among both CD4+ and CD8+ cells in mice with MAIDS were significantly higher than those for uninfected mice. These abnormalities were associated with presence of the MAIDS-inducing defective virus genome, but were not induced by infection of mice genetically resistant to development of MAIDS or with nonpathogenic helper murine leukemia virus, a virus component that induces high spontaneous proliferation of T cells, even in MAIDS-resistant mice.


Effects of CD4 synthetic peptides on HIV type I envelope glycoprotein function. Benzylated derivatives of a peptide (CD4(81-92)) representing the CDR3-like region of CD4 were previously found to inhibit gp120 binding, HIV-1 infectivity, and syncytium formation. These results have been interpreted to indicate a role for the corresponding CD4 region in these processes. The peptide (TbYICbEbVEDQKAcEE) is the prototype of a series of similar CD4(81-92) derivatives. We report that this peptide noncompetitively inhibits binding to CD4 of both gp120 and a mAb (MAX.16H5), both of which recognize the CDR2-like region of CD4. The binding of an antibody (Leu 3a) that is directed against a different area of the D1 domain of CD4 was also inhibited. The peptide derivative inhibited both HIV-1- and HTLV-1-mediated syncytium formation in the same concentration range. Nonbenzylated cyclic and linear peptides representing the CDR3-like region of CD4 (CD4(84-101)) had only minor effects on gp120 binding which were not sequence specific. The results of this study suggest that the effects of benzylated CD4(81-92) derivatives on HIV-1 binding or fusion should not be used to reach conclusions about the function of the corresponding CD4 region.


[A surgical case of thoracic aortic aneurysm due to Takayasu's aortitis associated with ulcerative colitis]. We experienced a case of thoracic aortic aneurysm due to Takayasu's aortitis associated with ulcerative colitis. Steroid was medicated to control inflammation and operation was performed. The ascending aorta, aortic arch, brachiocephalic artery, and left carotid artery were replaced by artificial graft. We made elephant trunk type anastomosis at the distal side of the graft to provide for growth of the aneurysm after operation. This was a rare case considered autoimmune overlapping syndrome, and its background was complicated by HLA-Bw52 and parasitic Metagonimus Yokogawai. Relationship between steroid medication and progression of the disease is not certain yet. Postoperative course is uneventful, no recurrence of inflammation is seen and the aneurysm is not enlarged until now.


Detection of bcr/c-abl mRNA in chronic myelogenous leukemia by polymerase chain reaction to identify chromosome translocation. The hallmark of chronic myelogenous leukemia (CML) is the Philadelphia chromosome (Ph1) which is caused by a translocation of the c-abl gene from chromosome 9 to the breakpoint cluster region (bcr) on chromosome 22. Polymerase chain reaction (PCR) can be used to detect the chimeric bcr/c-abl mRNA as evidence of the translocation. We applied a very simple and quick method of isolating cytoplasmic RNA, as well as reverse transcription and PCR in detecting bcr/c-abl mRNA of seven CML patients in various stages. Our results showed that only a small amount of either peripheral blood or bone marrow material was required for the expression of the bcr/c-abl mRNA. This is a very fast and time-saving method since a one-step method of cytoplasmic RNA isolation is used instead of the several steps in total RNA isolation as published in other literature.


Investigation into allergic response in patients with chronic sinusitis. We attempted to investigate the role of nasal allergy in sinusitis to elucidate whether it results from an immediate-type allergic reaction of the sinus mucosa or from allergic edema-induced sinus ostial obstruction. Forty-two patients with chronic sinusitis were selected for allergen skin tests, measurements of serum total and specific IgE, and sinus tissue-specific IgE. The data were then correlated to examinations of nasal mucosal scrapings and histopathology of the sinus mucosa. We found that serum levels of total IgE and house dust mite-specific IgE antibodies were significantly higher in patients (n = 12) allergic to house dust than in the nonatopics (n = 30; p less than 0.0001). There was no difference in the sinus tissue-specific IgE antibody. Eosinophils and basophilic cells in epithelial scrapings from the inferior turbinates, assessed by Hansel staining, were high in 66.7% and 50% of the atopic patients, respectively, and 36.7% and 26.7% of the nonatopics, respectively. The rates were influenced by the existence of infection and nasal polyps. The increase in eosinophils, mast cells and plasma cells, assessed by histopathologic examination, were not prevalent in the sinus mucosa of atopic patients. It is concluded that nasal allergy may be a predisposing factor to sinusitis and that the pathologic change of the sinus mucosa is mainly secondary, due to sinus ostial obstruction.


Changes in activity of cytochrome oxidase in the cochleae of guinea pigs with experimental endolymphatic hydrops. Cochlear dysfunction may indicate inadequate generation of intracellular metabolic energy which is necessary for the regulation of the transport of ions and fluid as well as production of electro-energy in the cochlea. Changes in cochlear function in the early stages of endolymphatic hydrops attributed to dysfunction of the sensory hair cells and stria vascularis were investigated. The main source of metabolic energy, produced by intracellular respiration, is located in the stria vascularis and sensory hair cells. Since cytochrome oxidase is one of the most important respiratory enzymes, vibratome sections of hydropic and normal cochleae were stained cytochemically for cytochrome oxidase activity in this study. Decreased activity of this enzyme was consistently shown in the normal-appearing outer hair cells and stria vascularis, as well as the degenerated hair cells and stria vascularis, of the higher turns of the hydropic cochlea. The results coincide with those in other studies of electrophysiologic changes in cochlear function in hydropic animals. A decrease in the activity of respiratory enzymes was noted before the destruction of the cellular structures. The activity of cytochrome oxidase may serve as a useful indicator for demonstrating the functional status of cochlear hair cells and stria vascularis.


Diurnal variation of insulin sensitivity in NIDDM patients and normal subjects. A modified insulin suppression test was adopted to assess the diurnal variation in insulin sensitivity and insulin clearance in 14 non-insulin-dependent diabetes mellitus (NIDDM) patients and eight age-, sex- and weight-matched normal subjects. The modified insulin suppression test was combined with an infusion of regular insulin, 30 mU/min x m2; glucose, 6 mg/kg x min; and somatostatin, 500 micrograms/h, for 120 minutes followed by only a somatostatin infusion for 60 minutes. Blood samplings were performed at appropriate times to obtain data on steady-state plasma insulin (SSPI), steady-state plasma glucose (SSPG as an index of insulin sensitivity), metabolic clearance and the half disappearance time (T1/2) of insulin. Blood specimens were also obtained during SSPI for measurement of erythrocyte insulin receptor binding. Each subject took the insulin suppression test twice. One test was started at 8 am and the other at 4 pm; each test was preceded by 16 hours of fasting. The order of the insulin suppression tests in each subject was randomized and balanced. In normal subjects, the SSPG level was lower in the morning than in the afternoon (118.0 +/- 43.6 vs 150.3 +/- 34.2 mg/dL, p less than 0.05). The NIDDM patients had a higher SSPG in the morning (217.7 +/- 51.4 vs 188.3 +/- 40.6 mg/dL, p less than 0.01). There was no diurnal difference in insulin clearance or the T1/2 in either normal subjects or NIDDM patients.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of tocopherol on platelet aggregation in non-insulin-dependent diabetes mellitus: ex vivo and in vitro studies. Tocopherol has been shown to have antiplatelet effects in insulin-dependent diabetes mellitus. However, its antiplatelet effect in non-insulin-dependent diabetes mellitus (NIDDM) remains to be established. In this report, the antiplatelet effect of tocopherol was assessed in a randomized, double-blind and crossover study of 15 NIDDM subjects. Each subject received tocopherol (dl-alpha-tocopherol nicotinate, 200 mg, tid) and a placebo for two six-week treatment periods separated by a three-week period in between for wash-out. The mechanisms of the antiplatelet effect of tocopherol were also studied in vitro. A significant decrease in platelet reactivity was observed after tocopherol treatment as compared with the pretest, and the magnitude of the decrease during tocopherol treatment was significantly evident when compared with that of the placebo treatment, as assessed by collagen (5, 10 micrograms/mL)-induced platelet aggregation of whole blood. A dose-dependent reduction in both ADP-and collagen-induced platelet aggregation was observed with tocopherol from 0.1 to 3.0 mM in vitro. No corresponding changes in ATP secretion and thromboxane synthesis were observed. Tocopherol also significantly inhibited fibrinogen-induced aggregation of elastase-treated platelets at a concentration of 0.1 mM. We demonstrated that platelet aggregation of whole blood ex vivo, among 15 NIDDM subjects was suppressed in tocopherol treatment, so tocopherol may have an antiplatelet effect in NIDDM subjects. The inhibitory effect of the platelet aggregation of tocopherol may be partially accomplished through interference with fibrinogen binding towards its receptor.


Prenatal cytogenetic diagnosis in amniocentesis. From 1982 to 1990, cytogenetic studies were successfully conducted in 2,975 (96.19%) of the 3,096 pregnant women who underwent amniocentesis. The average maternal age was 33.7 years and the average gestational age was 18.1 weeks. Common indications of amniocentesis included advanced maternal age (AMA) (54.76%), previous fetus with chromosomal aberrations (6.82%) or gross anomalies (5.01%), intrauterine gross anomaly (4.97%) and maternal exposure to drugs or radiation (5.28%). Among the 89 cases (2.99%) with detected chromosomal aberrations, 53 were numeric (31 trisomies, 21 sex chromosome aberrations and one tripoidy) and 36 were structural (six de novo and 30 hereditary structural rearrangement). The incidence of chromosomal aberrations was 2.03% in cases with AMA. While only four of the 143 cases with previous fetal trisomy 21 had recurrence, the recurrent rate was 90.91% in 11 cases with previous fetal chromosomal translocation. Thirty (20.27%) of the 148 cases with abnormal sonograms showed chromosomal aberrations. Certain congenital anomalies are closely associated with cytogenetic changes: duodenal atresia and trisomy 21; cystic hygroma and 45,X; and polyhydramnios and trisomy 18. Only two of the 157 cases with indications of drug or radiation exposure had abnormal cytogenetic studies. Two of the 53 cases with detected numerical aberrations (47,XXY and 47,XXX) and 27 cases with hereditary structural rearrangement elected to continue their pregnancies. All of these babies were delivered without gross anomalies. This study suggests that for prenatal diagnosis. However, complementary measures, such as routine antenatal ultrasound and maternal serum alphafetoprotein, should be added to increase the efficacy of genetic amniocentesis.


Serum gastrin levels in children with peptic ulcer disease. Fasting serum gastrin values were measured by radioimmunoassay in 53 children with peptic ulcer disease. The mean fasting serum gastrin levels for children with chronic duodenal ulcer, acute gastric ulcer, acute duodenal ulcer who were younger than two years old and acute duodenal ulcer who were older than two years old were: 46.2 +/- 25.7 pg/mL, 46.9 +/- 43.5 pg/mL, 47.9 +/- 12.0 pg/mL and 37.5 +/- 17.8 pg/mL, respectively. Children with peptic ulcer disease did not have elevated fasting serum gastrin levels when compared with age-matched controls. We conclude that a fasting serum gastrin value cannot be used as a screening test for peptic ulcer disease in children, except for Zollinger-Ellison syndrome.


Breast milk beta-glucuronidase in breast milk jaundice. Breast milk jaundice (BMJ) is the most common etiology of prolonged unconjugated hyperbilirubinemia in a newborn infant. The pathogenesis of BMJ has been studied by many investigators, yet the etiology still remains uncertain. A prospective study was done to examine the role of breast milk beta-glucuronidase in BMJ in healthy full-term Chinese newborns. There were 71 infants in the formula-fed group (FF group), 28 in the breast-fed group (BF group) and 27 in the mixed feeding group (MF group). Study infants were followed at the well-baby clinic for at least two months. Enzyme activity in maternal breast milk and infants' stools decreased significantly during the first week postpartum. There was no intergroup difference in fecal enzyme activity for the different age groups. Significantly more infants in the BF group (35.7%) developed prolonged unconjugated hyperbilirubinemia than did infants in the FF group (2.8%) and the MF group (0%). We did not find higher enzyme activity in breast milk or stools of infants with BMJ than in infants without BMJ. We, therefore, conclude that breast milk beta-glucuronidase is not related to the development of BMJ.


Comparison of different ligature materials used for T-tube esophageal exclusion. Four different ligature materials--plain catgut, chromic catgut, dexon and silk--were used for ligature of the distal arm during T-tube exclusion of the cervical esophagus in 12 dogs. Ligature by plain catgut was maintained for only a short period, but the duration of esophageal occlusion with the other three ligature materials was around 10 days. Ligated esophageal segments were examined grossly and histologically two months after the procedure. The diameter of the esophageal lumen in the ligated segments had become smaller compared with the neighboring normal esophageal lumen. The most prominent histologic changes were atrophy and fibrosis of the muscle coat, vessel congestion and inflammatory cell infiltration in the ligated segments. These tissue reactions were more severe in the chromic catgut and silk ligatures. Among the 11 evaluable dogs, four had symptoms of dysphagia after removal of the T-tube. All four dogs had a sinus discharge and granuloma formation at the T-tube esophagostoma. The diameter of the esophageal lumen was more constricted in dogs with dysphagia. Among the four ligature materials, dexon had the advantages of a long duration of occlusion, less tissue fibrosis and little sequel of esophageal stenosis, making it the most suitable for ligature during esophageal exclusion.


Meningeal carcinomatosis from solid tumors: clinical analysis of 42 cases. From January 1984 to June 1990, we observed 42 patients with meningeal carcinomatosis, 20 men and 22 women, aged 21 to 80 years (median age, 53 years). The two most common primary malignancies were lung cancer (50%) and breast cancer (31%). Sixty-four per cent was adenocarcinoma. On the first lumbar puncture, 86% had malignant cells in the cerebrospinal fluid. The findings of brain computed tomography were hydrocephalus (62%), contrast enhancement in the cerebral sulci or basal cisterns (31%), concomitant parenchymal metastases (15%) and normal scan (18%). In five out of seven cases, myelography showed irregular filling defects over the spinal cord or cauda equina. Treatment results were evaluated in 24 patients. Eight received radiation therapy (RT) alone, and 16 had combined therapy with RT plus intrathecal methotrexate (IT MTX). Of the patients who received RT alone, only one patient with lung carcinoma was stabilized clinically. Of the cases receiving combined therapy, seven improved clinically. Six of these were patients with breast carcinoma who received IT MTX via Ommaya reservoir. The latter had a median survival of 23 weeks. The follow-up period of the entire group of patients ranged from one day to 50 weeks. The median survival was four weeks. Based on this study, combined therapy with RT and IT MTX is indicated for breast carcinoma with meningeal carcinomatosis, but the therapeutic effects are uncertain for lung carcinoma and other malignancies.


Calcaneocuboid joint deformity in clubfoot. In the management of congenital clubfoot, recent attention has been given to the deformity of the calcaneocuboid joint in addition to the changes in the talonavicular, talocalcaneal and ankle joints. Fifty-five patients with 86 clubfeet operated on in the past six years were retrospectively studied. Radiographically, the calcaneocuboid subluxations were graded from I to IV by the Thoemtz classification. The grading has some correlation with the gross deformity, although there is no statistical data to confirm it. Postoperatively, those with grade I feet seemed to have satisfactory results with a posterior release only or with a posteromedial release. Grade II-III feet generally required extensive posteromedial and lateral release. Those who had a calcaneocuboid release and reduction obtained better correction than those who did not. We conclude that calcaneocuboid subluxation should be appraised preoperatively and reduced during surgery in order to achieve good results.


Results of Senning operation for transposition of great arteries. We report our results with 10 infants and children who underwent atrial repair using the Senning operation between 1985 and 1990. All cases had abdominal situs solitus, levocardia and atrio-ventricular concordance (D-bulboventricular loop). Nine patients had simple D-transposition of the great arteries without ventricular septal defects (VSD) or left ventricular outflow tract obstruction. The other patient had a double outlet right ventricle with subpulmonic VSD and pulmonary hypertension (PHT) and underwent a palliative Senning procedure. All patients had a balloon atrial septostomy (BAS) before surgery, except for one with Taussig-Bing syndrome. One patient had a Blalock-Hanlon operation after BAS. The age at the time of surgery ranged from two months to four years seven months (mean: 22 months) and weight ranged from 4.3-12 kg. There were two hospital mortalities including the patient with VSD and PHT. All of the patients had echocardiographic examinations and six of the eight survivors received cardiac recatheterization four to 19 months (mean: 7.4 months) postoperatively. No baffle leaks were noted in the survivors. Two patients had both mild tricuspid regurgitation and slightly decreased right ventricular contractility, and one patient had a pressure gradient of 6 mmHg between the superior vena cava and neo-right atrium. The clinical follow-up interval was eight to 64 months (mean: 31.4 months). All survivors showed a sinus rhythm on their latest electrocardiogram and were participating in normal daily activities without medication.