Myocardial function in immature and mature sheep with pressure-overload hypertrophy. The effect of pressure-overload left ventricular hypertrophy (LVH) on myocardial function is controversial. A major factor in the controversy may be the age at which the pressure overload was induced. The goal of this study was to determine whether the age at which the LVH was induced affected systolic myocardial function. We studied lambs (n = 8) and sheep (n = 7) with LVH induced by constricting the ascending aorta and their sham-operated controls (n = 7 and n = 7). This new, unsedated, ovine model of LVH was large enough to accommodate chronic surgical implantation of instrumentation and the induction of pressure-overload hypertrophy in both young and adult age groups. Solid-state intraventricular pressure transducers and sonomicrometer crystal were used to assess instantaneous left ventricular pressure and dimensions. Myocardial contractility was assessed with midwall shortening at common preload and afterload. Load alterations were induced by graded infusion of methoxamine. There was depressed myocardial function in adult sheep with LVH compared with adult controls. Lambs with and without LVH had normal left ventricular myocardial function, similar to adult controls. Similar results were obtained in studies after beta-adrenergic receptor blockade. We conclude that maturation decreases the ability of the myocardium to maintain normal contractility in the presence of pressure-overload hypertrophy.


Effects of a coronary alpha 1-constriction on transmural left ventricular flow and contractile function. Modulation of myocardial contractile function and perfusion by alpha 1-adrenergic receptors were examined in anesthetized dogs during left stellate ganglion stimulation. In 11 dogs, stellate stimulation significantly increased heart rate, mean arterial pressure, left ventricular systolic pressure, maximal rate of left ventricular pressure generation, segmental shortening and rate of shortening in anterior and posterior ventricular regions, and myocardial oxygen extraction. Myocardial lactate extraction decreased. The selective alpha 1-adrenergic antagonist prazosin (0.5 mg) injected into the circumflex artery during stellate stimulation caused significant additional increases in maximal rate of left ventricular pressure generation by 19 +/- 5% and in rate of shortening in posterior subendocardium by 20 +/- 6%. No changes were observed in posterior subepicardial or anterior subendocardial segmental contractile function. Myocardial oxygen and lactate extractions returned to their control values following prazosin injection. Regional left ventricular perfusion was measured using tracer microspheres in five additional dogs. Stellate stimulation increased subepicardial and subendocardial perfusion by 30%. Prazosin increased both subepicardial and subendocardial perfusion by an additional 36%. Stellate stimulation increased norepinephrine concentration in the coronary sinus, but no further increase was noted after blockage of alpha 1-receptors by prazosin. Thus, during sympathetic stimulation, an alpha 1-vasoconstriction existed uniformly across the left ventricular wall. However, blockade of this vasoconstriction was associated with an increase in contractile function only in the deeper muscle layers.


Hypothalamic excitatory amino acid receptors mediate stress-induced tachycardia in rats. The role of hypothalamic excitatory amino acid (EAA) receptors in mediating the cardiovascular response to stress was examined using conscious chronically instrumented rats. Microinjection of the EAA agonists N-methyl-D-aspartic acid (NMDA; 1-10 pmol), alpha-amino-3-hydroxy-5-methyl-4-isooxazolepropionic acid (AMPA; 0.3-3.0 pmol), or kainic acid (0.1-1.0 pmol) into the dorsomedial hypothalamus (DMH) elicited dose-related increases in heart rate and modest elevations in arterial pressure. Local microinjection of the NMDA antagonist 2-amino-5-phosphonopentanoic acid (AP5; 100 pmol) selectively blocked NMDA-induced cardiovascular changes, whereas the non-NMDA EAA antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 50 pmol) selectively blocked the responses to AMPA and kainic acid. In the stress trials, microinjection of the nonselective EAA antagonist kynurenic acid (1-10 nmol) into the DMH blocked air stress-induced tachycardia in a dose-related manner. Similar injection of kynurenic acid at sites lateral or posterior to the DMH or injection of xanthurenic acid (a structural analogue of kynurenic acid with no antagonistic properties at EAA receptors) into the DMH failed to influence air stress-induced cardiovascular changes. Injection of either AP5 or CNQX into the DMH at doses shown to be selective for their respective EAA receptor subtypes also attenuated air stress-induced tachycardia. Thus activity at EAA receptors in the DMH appears to be necessary for the generation of stress-induced changes in heart rate.


Large-cell acanthoma. A distinctive keratosis. Large-cell acanthoma is an epidermal neoplasm that is clinically, histologically, and biologically distinctive. Clinically, it differs from solar lentigo by being frequently skin-colored or hypopigmented. Histologically, it is defined by a population of uniformly large keratinocytes; it differs from solar lentigo by the absence of elongated hyperpigmented and sometimes hockey stick-shaped buds of keratinocytes. Biologically, it consists of hyperploid keratinocytes, whereas solar lentigo consists of diploid keratinocytes. Although the exact nosologic status of this entity is still controversial, its features are distinctive enough for the term "large-cell acanthoma" to merit continued usage.


Betamethasone-induced resistance to neuromuscular blockade: a comparison of atracurium and vecuronium in vitro. Steroids induce resistance to neuromuscular blocking drugs. Betamethasone-induced resistance to vecuronium has been demonstrated in vitro, and a presynaptic site of interaction has been suggested. This study investigated whether atracurium is similarly affected. Rat phrenic nerve-hemidiaphragm preparations were bathed in a physiologic solution, and one-half were exposed to betamethasone (1 mumol/L). Dose responses were recorded for atracurium (8-13 mumol/L) and vecuronium (2-12 mumol/L) for control and betamethasone-treated preparations. In comparison to control, the betamethasone groups had significantly less depression of muscle contraction force at all concentrations of atracurium (P = 0.0004) and vecuronium (P = 0.002). The calculated ED50 (50% depression of muscle contraction force, expressed as mean +/- SEM) for atracurium was 8.83 +/- 0.62 mumol/L for controls and 11.19 +/- 0.54 mumol/L for betamethasone-treated preparations. The calculated ED50 for vecuronium was 4.72 +/- 0.41 mumol/L for controls and 6.84 +/- 0.66 mumol/L for betamethasone-treated preparations. Betamethasone therefore increased the ED50 for atracurium by 27% and vecuronium by 45%; however, the magnitudes of these differences were not significant (P = 0.74) between the neuromuscular blocking agents. These results indicate that betamethasone-induced resistance to nondepolarizing neuromuscular blockade affects both atracurium and vecuronium to similar degrees in vitro.


The antihypertensive and cardiac hemodynamic effects of nebivolol. Acute, subacute, and chronic treatment with nebivolol, a novel beta 1-selective adrenergic antagonist, significantly lowered systolic and diastolic blood pressure and heart rate in patients with essential hypertension. No orthostatism and bradycardia were reported. A comparison between a normal control group and 23 hypertensive patients revealed that the ratio between the preejection period (PEP) and the left ventricular ejection time (LVET) of the systolic time intervals (STI) was significantly increased in the hypertensive patients, owing to a prolongation of the PEPc (PEP corrected for heart rate). Treatment with nebivolol 5 mg once a day significantly improved the PEP/LVET, in acute conditions from 0.42 +/- 0.023 to 0.39 +/- 0.018, after one month of treatment from 0.40 +/- 0.013 to 0.36 +/- 0.013 and after one year of treatment from 0.41 +/- 0.012 to 0.36 +/- 0.010, owing to a significant shortening of the PEPc. There was no correlation between changes of blood pressure and changes of STI. Diastolic dysfunction is an early finding in hypertension and may well be reflected in the prolongation of the PEPc. The improvement of left ventricular performance, as measured with STI, suggests that treatment with nebivolol may favorably influence the underlying diastolic dysfunction and be of therapeutic value for hypertensive patients with left ventricular damage.


Takayasu's arteritis with collateral circulation from the right coronary artery to intracranial vessels--a case report. A forty-four-year-old woman with Takayasu's arteritis and involvement of the aortic arch and its main branches complained of precordial pain on effort. Exercise electrocardiograms revealed significant ST segment depression in leads II, III, aVF, and V. Coronary arteriograms demonstrated no stenosis. However, the right coronary arteriogram revealed collateral circulation arising from the sinus node artery to the bilateral vertebral arteries and the left internal carotid artery. The collateral circulation was considered to be an important route of blood flow supply to the brain and, at the same time, a cause of coronary steal syndrome and, consequently, of angina pectoris.


[Embryonal carcinoma in the cryptorchid testis: apropos a case]. We report a case of testicular embryonal carcinoma in a patient with bilateral cryptorchidism. The diagnostic and therapeutic aspects of this disease entity are discussed. Although scrotal ultrasound constitutes an effective diagnostic method, it does not provide pathognomonic images that permit determining tumor histology as shown in the present case. Treatment of this tumor type depends on the degree of tumor spread: for stage I: inguinal orchidectomy and stage II: orchidectomy+lymphadenectomy. The need for adjuvant chemotherapy in these cases is discussed. In our view, it must depend on each specific case.


The differential reactivity of cells of the melanocytic lineage with four monoclonal antibodies against IFN-gamma inducible molecules. The reactivity of four monoclonal antibodies (MAbs) directed against IFN-gamma inducible antigens with melanocytic cells was investigated in the course of local and systemic tumor progression of human malignant melanoma. Frozen sections of histologically defined melanocytic tissues at different stages of progression were stained with these MAbs using an indirect immunoperoxidase technique. The reactivity of MAbs Me15/B3 and Me15/F9, directed against two different epitopes of a 90-kDa molecule, was found to correlate with melanoma progression. Indeed, a significantly lower percentage of small than of advanced primary melanomas or metastases stained positively. A differential staining of nevocytic and dysplastic nevi was further observed for these two MAbs, which were also non-reactive with normal skin melanocytes. The reactivity of MAb Me14/D12, which identifies the intercellular adhesion molecule ICAM-1 and MAb Mel14/F12, directed against a 40-kDa molecule, was found to be independent of the Breslow thickness of primary melanomas. Both the latter MAbs stained a high proportion of nevocytic and dysplastic nevi. The co-expression of the surface molecules defined by MAbs Me14/D12, Me15/B3 and Me15/F9 in the course of melanoma progression was also analyzed. The frequency of this co-expression increased according to the Breslow thickness of primary melanomas. In addition, up to 100% of metastases, as opposed to 20% of dysplastic nevi, were found to be simultaneously stained by these three MAbs. It is therefore conceivable that high-risk melanocytic lesions might be identified by the use of a combination of MAbs directed against IFN-gamma regulated antigens.


Electroencephalographic sleep abnormalities in schizophrenia. Relationship to positive/negative symptoms and prior neuroleptic treatment. Polysomnographic abnormalities in schizophrenia are not well characterized and their associations with schizophrenic symptomatology have not been adequately assessed. To address these issues, we recorded electroencephalographic sleep in 20 drug-naive schizophrenics, 20 drug-free but previously medicated schizophrenics, and 15 normal controls. Drug-naive and previously medicated patients had significantly greater impairment of sleep continuity and shorter rapid eye movement latency when compared with controls. In the previously medicated group, findings were significantly influenced by duration of drug-free status. Rapid eye movement latency was inversely correlated with the severity of negative symptoms (r = -.52) but was unrelated to depressive symptoms. Slow-wave sleep did not differ between schizophrenic patients and normal controls and was unrelated to any clinical parameter. Mechanisms underlying the observed associations between rapid eye movement sleep abnormalities and negative symptoms in the acute phase of schizophrenic illness need to be explored.


Exclusion of linkage between the serotonin2 receptor and schizophrenia in a large Swedish kindred. Family, twin, and adoption studies suggest that genetic factors play an important role in the etiology of schizophrenia. Detection of single gene(s) involved in a higher susceptibility to a hereditary disease is possible with linkage analysis. The effects of serotonin2-receptor antagonists on symptoms of schizophrenia suggest that a mutation in the gene coding for this receptor subtype might be involved in the pathophysiology of this disease. Recently a copy DNA encoding the serotonin 5-HT2 receptor has been isolated and with a human 5-HT2 receptor copy DNA probe the HTR2 locus has been mapped to chromosome 13. Using multipoint linkage analysis between schizophrenia and genetic markers spanning the region of the HTR2 locus, we were able to exclude linkage between this candidate gene and schizophrenia in a Swedish kindred. Given this result, we conclude that the serotonin 5-HT2 receptor gene itself is not a major susceptibility gene for schizophrenia in this family.


Oncoprotein (c-myc, c-erbB1, c-erbB2, c-fos) and suppressor gene product (p53) expression in squamous cell carcinomas of the lung. Clinical and biological correlations. The expression of the protooncogene encoded proteins (c-erbB1, c-erb B2, c-myc, c-fos) and the suppressor gene product p53 was analyzed in 81 human squamous cell carcinomas of the lung and correlated with clinical parameters of the patients (patient survival, presence of metastases and tumor stage) and with biological characteristics of the tumors (tumor growth in nude mice, DNA-ploidy, proliferative activity, drug-resistance and P-glycoprotein or gluathione S-transferase expression). By means of immunohistochemistry, expression of c-erbB1 oncoprotein (EGF-receptor) was detected in 79% of the tumors, c-erbB2 (c-neu) proteins in 35%, c-myc proteins in 48%, c-fos proteins in 41%, and p53 in 43% of the tumors. Patients with c-erbB1 positive tumors had a poor prognosis (p = 0.021). In addition, these tumors were more frequently drug resistant (p = 0.0067). A significant correlation between the growth of the squamous lung carcinomas in nude mice and c-fos oncoprotein expression was demonstrated (p = 0.017). Therefore, EGF-receptor and c-fos products may serve as prognostic factors for the aggressiveness of squamous cell carcinomas of the lung and for the response of these tumors to chemotherapy. No significant correlation was found between the expression of the c-erbB1 or c-fos gene products and stage, metastasis and DNA-ploidy. In contrast to these results, no relationship was found between c-neu or c-myc gene products expression and any of the clinical or biological parameters examined. Aneuploid squamous cell carcinomas of the lung expressed p53 more frequently than diploid tumors (p = 0.027). However, there was no significant difference between p53 expression and stage, survival of patients, metastasis, growth of the tumors in nude mice, proliferative activity and drug-resistance of the tumors.


Glucose attenuation of paradoxical sleep deficits in old rats. Glucose administration enhances memory in several amnestic populations, including old humans and rodents. The present experiment demonstrates that glucose also enhances measures of sleep in old rats. Three-hour day-time sleep EEGs were assessed in 3- and 24-month-old rats. The animals received injections of saline or glucose (100, 500, and 1000 mg/kg) on different days in a counter-balanced order. At doses of 100 and 500 mg/kg, glucose augmented the duration of paradoxical sleep bouts and total paradoxical sleep time in old, but not young, rats. Within 2 weeks after the sleep tests, measures of several brain neurotransmitter functions were obtained. Glucose was more effective in enhancing paradoxical sleep in those individual aged rats with high levels of hippocampal choline acetyltransferase and occipital cortex serotonin concentrations than in aged rats with lower levels on these neurochemical measures. The findings suggest that glucose attenuates selective age-related sleep deficits in old rats. More generally, these results add to a growing body of evidence indicating that moderate doses of peripheral glucose can influence a variety of CNS measures.


Lack of expression of P-glycoprotein in 7 small cell lung cancer cell lines established both from untreated and from treated patients. The establishment and characterisation of 7 small cell lung cancer cell lines is described. Four cell lines were established from biopsies taken from untreated patients and one of these was from primary tumour taken via the fibreoptic bronchoscope. The other three were from biopsies taken from patients who had relapsed after chemotherapy. All grow as non-adherent aggregates of cell and express a range of neuroendocrine and epithelial features characteristics of small cell lung cancer cell lines. No relationship was observed between the characteristics of the cell line in vitro and the treatment status of the patient from whom the biopsy was taken. Furthermore, none of the cell lines expressed p-glycoprotein even though 3 were derived from relapse biopsies where chemotherapy included drugs associated with the multidrug resistance phenotype.


Regulation of cardiac adenylate cyclase activity in rodent models of obesity. We have investigated beta-adrenergic regulation of adenylate cyclase activity in heart tissue membranes from the genetically obese Zucker rat, the genetically obese CBA mouse and the genetically obese diabetic (db/db) mouse. Responsiveness to beta-adrenergic stimulation was impaired in membranes from the obese Zucker rat, but not in the other models. The membranes from obese Zucker rats showed both decreased beta-adrenergic-receptor number and altered coupling between beta-adrenergic receptors and the stimulatory guanine-nucleotide-binding protein, Gs. In contrast, no alterations in either the levels of Gs or the functional interaction between this protein and the catalytic moiety of adenylate cyclase were observed. In these three genetic models of obesity we observe dissimilar alterations in the control of adenylate cyclase.


Regulation of the expression of the mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase gene. Its role in the control of ketogenesis. We have explored the role of mitochondrial 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase in regulating ketogenesis. We had previously cloned the cDNA for mitochondrial HMG-CoA synthase and have now studied the regulation in vivo of the expression of this gene in rat liver. The amount of processed mitochondrial HMG-CoA synthase mRNA is rapidly changed in response to cyclic AMP, insulin, dexamethasone and refeeding, and is greatly increased by starvation, fat feeding and diabetes. We conclude that one point of ketogenic control is exercised at the level of genetic expression of mitochondrial HMG-CoA synthase.


Coenzyme A is a potent inhibitor of acetyl-CoA carboxylase from rat epididymal fat-pads. Rat epididymal fat-pad extracts have previously been shown to contain an insulin-stimulated acetyl-CoA carboxylase kinase, which is co-eluted from Mono Q ion-exchange chromatography with a potent inhibitor of acetyl-CoA carboxylase [Borthwick, Edgell & Denton (1990) Biochem. J. 270, 795-801]. A variety of tests, including reactivity with thiol reagents, identify this inhibitor as CoA. Inhibition requires the presence of MgATP, but is independent of any phosphorylation of the enzyme. The effect is complete in about 5 min and is associated with depolymerization of acetyl-CoA carboxylase. Half-maximal inhibition is observed at about 40 nM-CoA. The inhibitory effects of CoA can be partially reversed by incubation with citrate and more fully overcome by treatment of the enzyme with the insulin-stimulated acetyl-CoA carboxylase kinase.


Cross-talk between M2 muscarinic and D1 dopamine receptors in the cat adrenal medulla. In the study reported here we have reached two conclusions. First, the cat adrenal medulla chromaffin cell possesses a dopamine D1 receptor that seems to be coupled to an adenylyl cyclase. Second, this receptor regulates the muscarinic-mediated catecholamine release response through a negative feed-back loop which uses cyclic AMP as a second messenger. These conclusions are supported by the following findings: (i) SKF38393 (a selective D1 receptor agonist), but not quinpirole (a selective D2 agonist), inhibits the methacholine-mediated catecholamine release responses in a concentration-dependent manner (IC50 of around 1-2 microM). (ii) SCH23390 (a selective D1 antagonist), but not sulpiride (a selective D2 antagonist), reversed by 70% the inhibitory effects of SKF38393. (iii) Dibutyril cyclic AMP (500 microM) inhibited by 80% the secretory effects of methacholine.


Lineage and stage specific expression of HOX 1 genes in the human hematopoietic system. Recent observations have demonstrated the expression of several members of the homeobox-containing (HOX) gene complexes within the hematopoietic compartment. We have analyzed the expression pattern of the entire HOX 1 locus in a panel of leukemia-derived human cell lines representing various blood phenotypes. The expression of the eleven HOX 1 genes is lineage-restricted and these genes are predominantly detected within cells of myelomonocytic origin. This is in strong contrast with the erythro-megakaryocytic specific expression of HOX 2 genes. Furthermore, we have observed that the expression of three HOX 1 genes within B lymphoid lineages is stage-related and that the expression of several of them is switched off during TPA-induced differentiation of Kg1 and U937. These observations suggest that HOX 1 homeoproteins could be regulators of lineage determination during hematopoiesis.


Mutations in the P53 tumour suppressor gene in primary lung cancer in Japan. The reverse transcription-polymerase chain reaction-single strand conformation polymorphism (RT-PCR-SSCP) analysis and sequencing were used to examine p53 gene alterations in 18 surgical specimens of primary lung cancers obtained in Japan. Somatic mutations resulting in amino acid changes were found in eight of the 18 cases (44%). Seven missense mutations were located in amino acid-conserved domains or their vicinities (codons 110 to 307). Most mutations were found at G-C pairs, suggesting that specific carcinogens are involved in the etiology of lung cancer. The p53 mutations showed a significant association with a history of smoking (P = 0.0294). We suggest that the p53 mutations may be associated with smoking-induced lung carcinogenesis.


Genomic differences between O6-methylguanine-DNA methyltransferase proficient (Mex+) and deficient (Mex-) cell lines: possible role of genetic and epigenetic changes in conversion of Mex+ into Mex-. Cell lines that possess O6-methylguanine-DNA methyltransferase (MGMT) repair activity (Mex+ phenotype) or are deficient for MGMT (Mex-) are compared at genomic level. It is shown that 1) 208F rat cells do not contain the MGMT gene, as detected by Southern blot hybridization. 2) Mex- HeLa MR and CHO-9 cells express very low amounts of MGMT mRNA, as detected by PCR. The size of the MGMT specific PCR product was slightly smaller than that generated from Mex+ HeLa S3 cells. 3) HeLa MR, compared to various human Mex+ cell lines, shows a restriction fragment length polymorphism indicating mutational alteration of MGMT gene sequences. 4) Mex- cells (HeLa MR) and cells that express very low MGMT activity (GM637) exhibit hypomethylation of the MGMT gene as revealed by MspI/HpaII restriction digests. 5) Exposure of Mex- cells to 5-azacytidine and selection with N-hydroxyethyl-N-chloroethylnitrosourea (HeCNU) did not yield Mex+ revertants. With V79 cells treated with 5-azacytidine clones resistant to HeCNU were isolated. These cells were MGMT deficient and not cross-resistant to N-methyl-N'-nitro-N-nitrosoguanidine indicating the existance of a defence mechanism other than MGMT against chloroethylating agents. The data suggest down-regulation of MGMT transcription accompanied by decreasing CpG methylation, but in some cell lines also mutational alterations to be involved in extinction of the Mex+ phenotype.


Apolipoprotein B genetic polymorphisms in several human hepatoma derived liver cell lines. The genetic polymorphism of apoB EcoRI and XbaI restriction sites and the 3' VNTR hypervariable region was examined in nine human hepatoma derived liver cell lines and related to the cells' ability to secrete lipids and apoB. EcoRI and XbaI genotypes appeared to be unrelated to triglyceride, cholesterol and apoB accumulating in the medium. The VNTR consisted of alleles with 47 to 67 repeats; however, these repeats were not associated with elevated concentrations of lipid or apoB. Data suggest that in the hepatoma cell lines, apoB polymorphisms in EcoRI, XbaI and the VNTR hypervariable region are not sufficient in themselves to account for triglyceride, cholesterol and apoB in the medium. It is possible that intracellular apoB synthesis and/or degradation as well as postsecretory apoB binding and uptake are responsible for the variability of apoB and lipid accumulation in the culture medium.


Partial amino acid sequence of a somatostatin receptor isolated from GH4C1 pituitary cells. A somatostatin receptor isolated from GH4C1 rat pituitary tumor-derived cells was cleaved with cyanogen bromide or cyanogen bromide+trypsin to obtain sequenceable fragments. Five unique amino acid sequences ranging from 6 to 27 amino acid residues were obtained. The sequence was identical to sequence recently reported for one of two somatostatin receptors cloned from human pancreas [Yamada et al., (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 251-255] except for a single valine to isoleucine substitution. This is the first report of amino acid sequence from a purified somatostatin receptor.


ADPRibosylation of chicken red cell tubulin and inhibition of microtubule self-assembly in vitro by the NAD(+)-dependent avian ADPRibosyl transferase. Chicken erythrocyte tubulin was found to undergo NAD(+)-dependent ADPribosylation in vitro in the presence of ADPRtransferase also isolated from avian red blood cells. Unlike the low level of ADPR incorporation catalyzed by Cholera and Pertussis toxins (i.e., less than 0.005 mol ADPR/mol tubulin), the avian system displayed a much higher stoichiometry of 0.8-1.2 mol ADPR/mol tubulin. Modification resulted in potent inhibition of microtubule self-assembly, even in the presence of bovine brain microtubule-associated proteins or with the addition of pre-assembled microtubules.


Piperine-mediated changes in the permeability of rat intestinal epithelial cells. The status of gamma-glutamyl transpeptidase activity, uptake of amino acids and lipid peroxidation. The effect of piperine (1-[5-(1,3-benzodioxol-5-yl)-1-oxo-2,4-pentadienyl]piperidin e), (from Piper nigrum) on the absorptive function of the intestine was studied. In vitro experiments showed that piperine (25-100 microM) significantly stimulated gamma-glutamyl transpeptidase (gamma-GT, EC 2.3.2.2.) activity, enhanced the uptake of radiolabelled L-leucine, L-isoleucine and L-valine, and increased lipid peroxidation in freshly isolated epithelial cells of rat jejunum. The kinetic behaviour of gamma-GT towards substrate and acceptor altered in the presence of piperine. In the presence of benzyl alcohol, an enhanced gamma-GT activity due to piperine was maintained. These results suggested that piperine may interact with the lipid environment to produce effects which lead to increased permeability of the intestinal cells.


Drug transport, viability and morphology of isolated rat hepatocytes preserved for 24 hours in University of Wisconsin solution. Isolated hepatocytes are a valuable tool to study liver functions. Suitable methods to preserve the isolated cells with good maintenance of viability and functions are crucial to extend experiments with hepatocytes from a single isolation over 2 or more consecutive days. We investigated whether University of Wisconsin (UW) solution, which was designed to preserve organs for transplantation, is also suitable for preservation of isolated rat hepatocytes. Viability, as determined by Trypan blue exclusion and reduction of the tetrazolium 3(4,5-dimethyl-thiazoyl-2-yl)2,5 diphenyltetrazolium bromide to a purple formazan, morphological appearance using electron microscopy, ATP levels and uptake and storage of three model drugs ([3H]vecuronium, [3H]taurocholic acid and [3H]ouabain) were determined directly after isolation and after 22 hr of storage in UW solution at 0-4 degrees. The present study shows that cold storage of rat hepatocytes for 22 hr in UW can be performed without significant loss of viability and with maintenance of proper morphology, cellular ATP and transport functions. In contrast, after storage in Krebs-Henseleit buffer the normal morphology, ATP content and transport functions were strongly affected. These results imply that hepatocytes from a single isolation and stored in UW solution can be used for experiments on 2 consecutive days.


Differences in T cell receptor restriction fragment length polymorphisms in patients with rheumatoid arthritis. The purpose of this study was to determine whether a T cell receptor (TCR) polymorphism, either by itself or in combination with particular HLA polymorphism, leads to susceptibility to rheumatoid arthritis (RA). Eight restriction fragment length polymorphisms (RFLPs) detected with TCR gene segments were investigated in 46 individuals with RA and were compared with data from normal control subjects. A statistically significant difference in the genotype frequencies of a Taq I RFLP detected with the TCR alpha constant region (C alpha) gene was noted. In addition, when the DR4+ subpopulations were examined, the allelic frequency of a 2-kb Bam HI fragment detected with a V beta 8 gene was increased in the samples from RA patients (P less than 0.0086). The results of this study suggest that germline differences in the TCR repertoire may be associated with RA, and that there is a contributory effect of DR4+ haplotypes with certain TCR haplotypes in susceptibility to RA.


Magnetic resonance imaging detection of aortic and pulmonary artery wall thickening in the acute stage of Takayasu arteritis. Improvement of clinical and radiologic findings after steroid therapy. Early diagnosis of Takayasu arteritis in the acute stage (prepulseless stage) is extremely difficult. Identification of a useful approach to detecting the initial changes of arteritis is therefore desirable. Careful clinical examination of a young woman with persistent fever and dry cough revealed faintly audible bruits at the cervical, supraclavicular, and abdominal regions. Aortographic features suggested thickening of the wall of the descending thoracic aorta. Magnetic resonance imaging (MRI) of this area was diagnostic. MRI demonstrated involvement of the ascending aorta and right main pulmonary artery. Steroid therapy (prednisolone 60 mg/day) induced dramatic clinical and radiologic improvement in 2 months. This is the first report of MRI-documented reduction in the thickness of the walls of both the aorta and the pulmonary artery following steroid therapy.


Role of glutathione peroxidase in rheumatoid arthritis: analysis of enzyme activity and DNA polymorphism. Aberrant expression of the antioxidant enzyme glutathione peroxidase (GPx) could contribute to the etiology of rheumatoid arthritis (RA). However, previous enzyme activity studies examining this relationship were inconclusive. Indirect evidence for this relationship derives from the known efficacy of gold therapy in RA, since gold compounds specifically inhibit GPx. The hypothesis that variants of GPx are associated with RA was examined by two approaches: enzyme activity analysis and restriction fragment length polymorphism (RFLP) association analysis. No significant difference was found in whole blood GPx activity between 28 RA patients and 36 controls. GPx activity appeared to be independent of sex, race, or type of drug treatment. However, a statistically significant difference was found with respect to treatment responsiveness. RA patients classified as good responders to gold therapy, but who were no longer taking gold, had a significantly higher GPx activity compared to both the controls and good responders currently on gold therapy. Aberrantly high GPx activity could contribute to RA by generating excess oxidized glutathione, a potent collagenase activator. Gold therapy would reduce GPx activity to normal levels. The restriction enzyme Pvu II in conjunction with a GPx gene probe identified a useful RFLP (Al, 22 kbp; A2, 15 kbp) with allelic frequencies of A1 and A2 equal to 0.11 and 0.89, respectively, in the control population. No statistically significant association, however, could be demonstrated between this allelic variant of the GPx gene and RA.


Correlation between kinetics of soluble CD4 interactions with HIV-1-Env-expressing cells and inhibition of syncytia formation: implications for mechanisms of cell fusion and therapy for AIDS. To study the kinetics of the interactions between soluble (s) CD4 and HIV-1-Env-expressing cells in relation to subsequent events leading to cell fusion and inhibition of syncytia formation. Vaccinia-HIV-1 (Env)-infected CD4- T-cells were used to study the kinetics of sCD4-gp120/41 interactions and syncytia formation (with CD4+ T-cells) under identical conditions. sCD4 association and dissociation rates for HIV-1-Env-expressing cells, and quantification of sCD4-induced gp120 shedding was determined by a quantitative flow cytometry assay. Syncytia inhibition was measured in the continuous presence of sCD4, or after washing of HIV-1-Env-expressing cells following pre-incubation with sCD4. The kinetics of syncytia inhibition correlated with sCD4 binding when sCD4 was maintained during the culture. When Env-expressing cells, which had been pre-incubated with sCD4, were washed to remove unbound sCD4, no syncytia formation inhibition was observed, even following sCD4-induced shedding of greater than 50% of surface gp120 molecules. The lack of syncytia inhibition seen after removal of unbound sCD4, even after pre-incubation of cells under saturation and gp120 shedding conditions, indicated that sufficient numbers of fusogenic molecules remained on the sCD4-treated cells. In addition, fast dissociation of pre-bound sCD4 occurred in culture. These results are important for understanding HIV-1-Env-mediated cell fusion and AIDS therapy.


Clinical features and predictive markers of disease progression in cynomolgus monkeys experimentally infected with simian immunodeficiency virus. To study the pathogenicity of simian immunodeficiency virus (SIVsm) in cynomolgus monkeys in order to establish an animal model for human AIDS. Thirty-three cynomolgus monkeys were monitored for more than 2 years following experimental infection with SIVsm. All the macaques became SIV-infected, as demonstrated by virus recovery from peripheral blood lymphocytes and by the appearance of viral antibodies. SIVsm was found to be pathogenic, killing 29 out of the 33 monkeys (88%) within 26 months. Clinically, infected monkeys developed lymphadenopathy, splenomegaly, diarrhoea, weight loss, neurological symptoms and a remarkably high incidence (39%) of malignant lymphomas. All lymphomas were high-grade malignant and of B-cell origin. Disease progression was associated with low CD4+ lymphocyte count, involution of initially hyperplastic follicular B-cell areas in lymph nodes, reappearance of viral antigen in serum, loss of anti-Gag antibodies and development of systemic giant cell disease in 55% of the monkeys. There are many similarities between SIVsm-induced AIDS in cynomolgus monkeys and human AIDS with regard to clinical, virological, immunological and pathological manifestations.


Impaired T-cell priming and proliferation in cats infected with feline immunodeficiency virus. Cats naturally infected with feline immunodeficiency virus (FIV) are particularly susceptible to infection with opportunistic pathogens, suggesting that these animals are unable to develop an effective immune response against the pathogen. Previous studies have used CD4+:CD8+ lymphocyte ratios and mitogen blastogenesis to identify immunological abnormalities in FIV-infected cats. However, these studies provide limited information for understanding the nature of the cellular dysfunction in FIV-infected cats, particularly defects in antigen-specific immune responses. To investigate whether cats infected with FIV are less able to mount an immune response to previously unencountered antigens, we compared the development of antigen-specific cellular immunity at the stage of T-cell priming in uninfected and FIV-infected cats. The general immune status of cats was assessed by peripheral blood CD4+:CD8+ lymphocyte ratios (flow cytometry), and by lymphocyte blastogenesis response to T- and B-cell mitogens. In addition, we describe the development of an autologous culture system to measure specific priming of naive feline T-cells to soluble antigen in vitro. This assay was used to compare T-cell priming in uninfected cats and cats which had been infected with FIV for 6-27 months. As in HIV infection, CD4+:CD8+ lymphocyte ratios in FIV-infected cats were found to be inverted, due to a reduction in the percentage of CD4+ cells. In addition, lymphocyte blastogenesis to both T- and B-cell mitogens was significantly impaired in FIV-infected cats. Priming to keyhole limpet haemocyanin (KLH) elicited a late proliferative response resulting from the expansion of CD4+ (T-helper cells). T-cell growth factor secretion correlated with cell proliferation. Restimulation of cells with fresh antigen-presenting cells and antigen showed that antigen-specific T-cell priming had occurred in the initial culture. When primary proliferation responses in FIV-infected cats were examined, it was observed that naive CD4+ T-cells from FIV-infected cats were significantly impaired (P less than 0.001) in their ability to be primed to KLH when compared with uninfected controls. Impaired priming of naive CD4+ T-helper cells to antigen in FIV-infected cats may explain the increased susceptibility of these animals to infection by opportunistic pathogens. The poor ability of human patients with AIDS to develop humoral immunity following vaccination may also be caused by such a defect in T-cell priming.


Diagnostic accuracy of three clinical case definitions for advanced HIV disease. To assess the accuracy of three clinical case definitions for advanced HIV disease: the World Health Organization (WHO) case definition, and the original and revised Caracas case definitions. Retrospective chart review. A clinic for patients with all stages of HIV infection at the Johns Hopkins Hospital, Baltimore, [correction of Bethesda] Maryland, USA, a tertiary care university hospital. Two hundred and twenty-four HIV-positive adults who underwent initial evaluation between 1 January 1990 and 31 December 1990. A score for each definition was assigned based on initial evaluation. The sensitivity, specificity, and predictive values were calculated using the Centers for Disease Control (CDC) staging criteria, and results were correlated with total CD4 cell counts. The sensitivities of the WHO, and the original and revised Caracas definitions were 40, 67, and 60%, respectively, using CDC disease stage IV as a positive standard. Specificities were between 99 and 100%, using CDC stage II-III disease as a negative standard. Mean CD4 cell counts for patients with positive scores were 184, 160, and 158 x 10(6)/l, respectively, compared to 191 x 10(6)/l for CDC stage IV patients. Sensitivity was lower when the positive standard was expanded to include all patients with CD4 cell counts less than 200 x 10(6)/l. In our study population, case definitions were specific, but only moderately sensitive for advanced HIV disease. Prospective studies should be conducted in diverse geographic regions, using lymphocyte or CD4 cell counts when possible.


Distribution of tyrosine hydroxylase immunoreactive nerve fibers in the canine larynx. The sympathetic innervation of the canine larynx was investigated using tyrosine hydroxylase (TH) immunohistochemistry. Many tyrosine hydroxylase immunoreactive (TH-IR) nerve fibers were observed around arteries and arterioles in the laryngeal mucosa and intrinsic laryngeal muscles. In the glandular region, TH-IR fibers were also found, with some of these fibers terminating around the basement membranes of the glandular cells. The quantity of TH-IR fibers in the mucosa differed among regions of the larynx. Many of these fibers could be found in the laryngeal surface of the epiglottis as well as the posterior glottis. These findings suggest that TH-IR fibers may directly innervate muscles in the intrinsic larynx.


Developmental switch in the pharmacology of Ca2+ channels coupled to acetylcholine release. The pharmacological specificity of Ca2+ channel-secretion coupling in acetylcholine (ACh) and somatostatin (SOM) release was studied in the chick eye choroid neuromuscular junctions and in dissociated ciliary ganglion (CG) neurons. ACh secretion changes in development from stage (St) 40, when release is dihydropyridine (DHP) and partially omega-conotoxin (omega-CgTX) sensitive, to posthatch, when release is insensitive to DHPs but sensitive to omega-CgTX. St 40 CG neurons cultured with striated muscle have release properties similar to those of St 40 iris and choroid but different from those of St 34 neurons, which are neither DHP nor omega-CgTX sensitive. SOM (also coreleased from posthatch choroid terminals) can inhibit ACh release in both posthatch and St 40 choroids, suggesting that the SOM receptor interacts with both DHP-sensitive and -insensitive channels.


Rostrocaudal polarity of the tectum in birds: correlation of en gradient and topographic order in retinotectal projection. Rostrocaudal polarity of the tectum is first detectable at embryonic day 2 (E2) in the chick by the rostrocaudal gradient of en expression. When ectopic tectum is produced by heterotopic transplantation at E2 in the diencephalon, the gradient of en expression is inverted from the host polarity by environmental influences. Here we report that the retinal fibers project to the ectopic tectum in a topographic order in accord with the inverted gradient of en expression. Moreover, if ectopic tectum was produced at E3, when the en pattern was preserved, the retinotectal projection pattern was also in accord with the en pattern, suggesting that rostrocaudal polarity of retinotectal order follows en expression patterns.


Inhibition of the omega-conotoxin-sensitive calcium current by distinct G proteins. Leu-enkephalin (Leu-Enk), norepinephrine (NE), somatostatin (SS), and bradykinin (BK) decrease the voltage-dependent calcium current in NG108-15 cells. Here we have investigated whether distinct G proteins, or a G protein common to all of the pathways, mediates this inhibition. We found that pertussis toxin (PTX) reduced all of these transmitter actions, except that of BK. To examine which of the PTX-sensitive pathways is transduced by GoA, we constructed an NG108-15 cell line that stably expresses a mutant, PTX-resistant alpha subunit of GoA. After treatment with PTX, the mutant GoA alpha rescued the Leu-Enk and NE pathways but not the SS pathway. At least three different G proteins can transduce receptor-mediated inhibition of calcium currents in nerve cells. The effects of these G proteins appear to converge on the omega-conotoxin GVIA-sensitive calcium current.


A cell line with unusual characteristics from an ovarian carcinoma patient: modulation of sensitivity to antitumour drugs. A cell line (GZL-8) was established by cloning from ascitic fluid of an untreated ovarian carcinoma patient. The cells grew rapidly, accumulated lipids and showed chromosomal alterations. One of the marker chromosomes showed characteristics of a Y-like chromosome. This unusual finding was confirmed by DNA hybridisation using specific probes to the Y chromosome. The cells stained with fluorescent antibodies to desmoplakin and cytokeratins 8, 18, 19, and weakly with vimentin but not with desmin. The presence of epithelial membrane antigen, human milk fat globulin, alpha-lactalbumin, alpha-fetoprotein, placental alkaline phosphatase and oestrogen receptor-related antigen was demonstrated by indirect immunoperoxidase staining, but no CA-125 antigen could be detected. The cells showed positive reaction with antibodies to P-glycoprotein. The function of the P-glycoprotein transport system was demonstrated by the rhodamine-123 release test. The cells were initially responsive to doxorubicin, and to high concentrations of cisplatin. Growth inhibition by doxorubicin, especially at low doses was enhanced by the addition of verapamil or tamoxifen. This was shown by the soft agar clonogenic assay, by direct cell counting and by the MTT reducing test. Our results show that combination between drug and sensitivity modulators may be of potential clinical value in ovarian cancer.


Identification and interpretation of epidermal growth factor and c-erbB-2 overexpression. Overexpression of normal cellular genes may be one mechanism by which malignant cells can acquire a selective growth advantage. The epidermal growth factor receptor and the c-erbB-2 protein are members of the erbB family and are good examples of genes that appear to act through this mechanism. Molecular and biochemical analyses of these two proteins also illustrate how studies of growth factors, growth factor receptors and oncogenic retroviruses may lead to new approaches to diagnosis and treatment. In particular, overexpression of these growth factor receptors has identified clinical subgroups that may respond differently to chemotherapy and provides the opportunity for antibody targeted therapy. Overexpression of these proteins can be identified using immunocytochemistry on both histological sections and fine-needle aspirates, thus enabling these parameters to be assessed preoperatively and to be monitored during therapy.


Conformational states of ribulosebisphosphate carboxylase and their interaction with chaperonin 60. Conformational states of ribulosebisphosphate carboxylase (Rubisco) from Rhodospirillum rubrum were examined by far-UV circular dichroism (CD), tryptophan fluorescence, and 1-anilino-naphthalenesulfonate (ANS) binding. At pH 2 and low ionic strength (I = 0.01), Rubisco adopts an unfolded, monomeric conformation (UA1 state) as judged by far-UV CD and tryptophan fluorescence. As with other acid-unfolded proteins [Goto, Y., Calciano, L. J., & Fink, A. L. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 573-577], an intermediate conformation (A1 state) is observed at pH 2 and high ionic strength. The A1 state has an alpha-helical content equivalent to 64% of that present in the native dimer (N2 state). However, fluorescence measurements indicate that the tertiary structure of the A1 state is largely disordered. A site-directed mutant, K168E, which exists as a stable monomer [Mural, R. J., Soper, T. S., Larimer, F. W., & Hartman, F. C. (1990) J. Biol. Chem. 265, 6501-6505] was used to characterize the "native" monomer (N1 state). The far-UV CD spectra of the N1 and N2 states are almost identical, indicating a similar secondary structure content. However, the tertiary structure of the N1 state is less ordered than that of the N2 state. Nevertheless, when appropriately complemented in vitro, K168E forms an active heterodimer. Upon neutralization of acid-denatured Rubisco or dilution of guanidine hydrochloride-denatured Rubisco, unstable folding intermediates (I1 state) are rapidly formed. At concentrations at or below the "critical aggregation concentration" (CAC), the I1 state reverts spontaneously but slowly to the native states with high yield (greater than 65%). The CAC is temperature-dependent. At concentrations above the CAC, the I1 and the A1 states undergo irreversible aggregation. The commitment to aggregation is rapid [ef. Goldberg, M. E., Rudolph, R., & Jaenicke, R. (1991) Biochemistry 30, 2790-2797] and proceeds until the concentration of folding intermediate(s) has fallen to the CAC. In the presence of a molar excess of chaperonin 60 oligomers, the I1 state forms a stable binary complex. No stable binary complex between chaperonin 60 and the N1 state could be detected. Formation of the chaperonin 60-I1 binary complex arrests the spontaneous folding process. The I1 state becomes resistant to interaction with chaperonin 60 with kinetics indistinguishable from those associated with the appearance of the native states. In vitro complementation analysis indicated that the product of the chaperonin-facilitated process is monomeric.(ABSTRACT TRUNCATED AT 400 WORDS)


Promotion of the in vitro renaturation of dodecameric glutamine synthetase from Escherichia coli in the presence of GroEL (chaperonin-60) and ATP. The folding and assembly of dodecameric glutamine synthetase (GS) from Escherichia coli was examined in the absence and presence of the E. coli heat shock protein, GroEL (chaperonin-60). At nonphysiological temperatures (15-20 degrees C), unfolded GS spontaneously renatured to 80-90% of its original activity in the absence of GroEL. At near-physiological temperatures (37 degrees C), only 20-40% of the original activity returns. Under the latter solution conditions, GroEL and ATP enhance the extent of GS renaturation to 70-80% of the original activity at 37 degrees C. In the absence of ATP, GroEL arrests the renaturation of unfolded GS by forming a stable binary complex. The addition of ATP to this complex resulted in the release of GS subunits and formation of active dodecameric GS. The order of addition of ATP or unfolded GS to GroEL results in differences in the t1/2 values where half-maximal GS activity is attained. At a constant GS concentration, the formation of the GroEL.GS complex followed by ATP addition resulted in approximately a 2-fold increase in the observed t1/2 value compared to that observed when GroEL was preincubated with ATP before the GS renaturation reaction was initiated. These differences in renaturation rates may be related to binding affinity differences between the ATP-free and -bound GroEL conformer for unfolded or partially folded protein substrates [Badcoe, I. G., Smith, C. J., Wood, S., Halsall, D. J., Holbrook, J. J., Lund, P., & Clarke, A. R. (1991) Biochemistry 30, 9195-9200].(ABSTRACT TRUNCATED AT 250 WORDS)


Early-life patterns of plasma gut regulatory peptide levels in calves: effects of the first meals. The effects of the first meals on the release of seven gut regulatory peptides were studied in newborn calves fed colostrum either at serial intervals during the first day of life or at 28 h only. Fasted animals showed no significant variation of plasma peptides until the first feed, except for somatostatin, which peaked at 4-5 h and declined thereafter. As assessed before and 1 h after feeding, the first meal tended to induce rises in plasma gastrin, cholecystokinin and pancreatic polypeptide, while the other peptides were unaffected. Repeated colostrum feeds induced marked increases in plasma gastrin, cholecystokinin, secretin and vasoactive intestinal peptide from 10 h on. Pancreatic polypeptide was transiently increased from 4 to 16 h. Feeding was followed by a transitory reduction of plasma somatostatin and by a prolonged decrease of plasma motilin. We conclude that colostrum feeding potently modulates the release of several regulatory peptides shortly after birth in calves. These responses may be important for the adaptation of gut growth, secretions and motility to food ingestion in the neonatal period.


Carcass composition and resistance to fasting in neonatal piglets born of sows immunized against somatostatin and/or receiving growth hormone-releasing factor injections during gestation. Thirty-eight gestating sows were either immunized against somatostatin (SRIF) and/or injected with growth hormone-releasing factor (GRF). Treatment effects on carcass composition and resistance of newborn piglets to a 60-hour fast were investigated. Protein content of carcasses at birth was increased in piglets of sows receiving GRF or immunized against SRIF, however, when sows received both treatments there was a reduction in carcass protein content (p = 0.01). Other carcass components were unaltered by treatments, and none of the treatments affected metabolic or endocrine profiles of piglets at birth. Concentrations of GH, IGF-I (p less than 0.01), glucagon and cortisol (p less than 0.05) increased linearly with duration of fast, whereas glucose values decreased. Resistance to fasting was unaltered in piglets from any treatment thereby suggesting that exogenous GRF and/or SRIF immunization of sows during gestation are unlikely to improve survival of newborn piglets.


A kinetic model of CD4+ lymphocytes with the human immunodeficiency virus (HIV). This report describes a kinetic model of in vitro cytopathology involving interactions of human immunodeficiency virus (HIV) with CD4+ helper T lymphocytes. The model uses nonlinearly coupled, ordinary differential equations to simulate the dynamics of infected and uninfected cells and free virions. It is assumed that resting cells are more readily infected than activated cells, but once infected, only activated cells produce more virus. Resting cells can be activated by some appropriate stimulus (e.g. phytohemagglutinin, soluble antigen). The model predicts that the initial inoculum of virus is taken up by resting cells and without stimulation the system comes to a steady state of two populations, namely infected and uninfected cells. Stimulation of this system produces two additional populations, namely infected and uninfected activated cells which, along with the previous populations, exhibit cyclic behavior of growth, viral expression/release, and death. Additional stimuli enhance or diminish the cyclic behavior depending upon their occurrence in time. These simulations suggest a similar dynamics in human HIV infection and may explain a major factor responsible for the widely varying depletion rate of (CD4+) helper T cells in AIDS patients.


Esmolol infusion during nitroprusside-induced hypotension: impact on hemodynamics, ventricular performance, and venous admixture. The impact of esmolol infusion on hemodynamics, ventricular performance, venous admixture, sympathoadrenal, and renin-angiotensin system responses during sodium nitroprusside (SNP)-induced hypotension was studied in 11 patients undergoing lymph node dissection during general anesthesia with 60% nitrous oxide and fentanyl. Radial arterial and thermistor-tipped pulmonary catheters were employed for hemodynamic monitoring. Arterial and mixed venous blood gas tensions, arterial plasma renin activity (PRA), and plasma catecholamine levels were measured. Derived hemodynamic parameters and venous admixture (Qs/Qt) data were obtained from standard equations. Transesophageal echocardiography (6 patients) was used to assess left ventricular performance using the relationship between end-systolic wall stress (ESWS) and velocity of circumferential shortening (VCFC). After surgical incision, arterial hypotension was induced with SNP alone. Esmolol was infused at each of the following rates in sequence: 200, 300, and 400 micrograms/kg/min. Each esmolol infusion lasted 20 minutes and the SNP dose was adjusted to maintain MAP at 55 to 60 mm Hg. The mean dose of SNP required to induce hypotension was 5.5 micrograms/kg/min +/- 0.5 SE. Compared to prehypotension values, SNP induced significant increases in Qs/Qt and reductions in PaO2, systemic vascular resistance (SVR), and stroke volume index (SVI). Esmolol infusion caused dose-dependent (highest with 400 micrograms/kg/min) reductions in the SNP requirement, heart rate (HR), SVI, Qs/Qt, and PRA, and also led to significant increases in SVR and left ventricular (LV) internal diameter in diastole as well as systole. Furthermore, esmolol infusion was associated with a dose-dependent downward and leftward shift of the ESWS versus VCFC relationship, implying diminished contractility.(ABSTRACT TRUNCATED AT 250 WORDS)


Pulmonary infiltration and eosinophilia associated with sulfasalazine therapy for ulcerative colitis: a case report and review of literature. We report a 52-yr-old man with ulcerative colitis who developed sulfasalazine-induced pulmonary infiltration with eosinophilia (PIE syndrome), which resolved completely after withdrawal of this drug. Desensitization to sulfasalazine was successful, and allowed the patient to receive this drug without recurrence of the pulmonary toxicity. This is the first case of the sulfasalazine-induced PIE syndrome in Japan; a review of the world literature found no previous cases of successful desensitization following sulfasalazine-induced PIE syndrome.


c-myc, c-erbB-2, and Ki-67 expression in normal breast tissue and in invasive and noninvasive breast carcinoma. c-myc, c-erbB-2, and Ki-67 expression was examined by immunohistochemistry in 11 normal breast tissues and 42 invasive and 14 noninvasive breast carcinomas. The c-myc product was detected in all breast carcinoma specimens and in 7 of 11 normal breast tissues. Invasive tumors stained more frequently with the anti-myc monoclonal antibody than did noninvasive tumors, while the level of expression in normal breast tissue was much less than that in breast cancer. Membrane staining of the c-erbB-2 protein was demonstrated in 29% (4 of 14) of noninvasive ductal carcinomas and in 45% (19 of 42) of invasive breast carcinomas. None of the 11 normal breast tissue samples was positive. The mean value of Ki-67-positive cells was 0.91 +/- 0.31% for normal breast tissue, 4.57 +/- 1.36% for noninvasive ductal carcinoma, and 12.76 +/- 2.18% for invasive breast cancer. In 42 invasive breast carcinomas, the expression of c-myc, c-erbB-2, and Ki-67 proliferation marker were compared with lymph node status, estrogen receptor status, progesterone receptor status, and age of patients at diagnosis. c-erbB-2 overexpression and Ki-67 overexpression were identified as the only factors associated with lymph node status. We concluded that they might be additional prognostic factors for breast carcinoma.


Soluble intercellular adhesion molecule 1 is released by human melanoma cells and is associated with tumor growth in nude mice. We have studied the cytokine regulation of cell surface and soluble intercellular adhesion molecule 1 (ICAM-1) expression on the human melanoma cell line A375M. Unstimulated cells express ICAM-1 on their cell surface but do not secrete significant levels of soluble ICAM-1. Interleukin 1, interleukin 6, tumor necrosis factor, and gamma-interferon all increased cell surface expression of ICAM-1. Tumor necrosis factor, interleukin 1, and gamma-interferon also caused the release of soluble ICAM-1. The serum of melanoma patients has been reported to contain elevated levels of soluble ICAM-1; however, the source of this ICAM-1 is unclear. The serum from nude mice bearing s.c. human melanoma tumors was found to contain soluble human ICAM-1. ICAM-1 levels showed a positive correlation with tumor weight. The release of ICAM-1 from melanoma tumors, in response to host-derived cytokines, may have relevance to immune recognition of the tumor.


Homeosis in the mouse induced by a null mutation in the Hox-3.1 gene. We have replaced the Hox-3.1 coding sequence with the E. coli lacZ gene by means of homologous recombination in embryonic stem cells and thus produced null mutant mice. Homozygous mice were born alive, but most of them died within a few days. In the trunk region of homozygotes, several skeletal segments were transformed into the likeness of more anterior ones, as observed in Drosophila with loss-of-function homeotic mutations. The most obvious transformations were the attachment of the 8th pair of ribs to the sternum and the appearance of a 14th pair of ribs on the 1st lumbar vertebra. The pattern of beta-galactosidase activity was identical in heterozygotes and homozygotes and reflected faithfully the Hox-3.1 expression pattern. Thus, the mutation modified the identity, rather than the position, of embryonic cells that would normally express Hox-3.1.


Structurally distinct and stage-specific adenylyl cyclase genes play different roles in Dictyostelium development. We have isolated two adenylyl cyclase genes, designated ACA and ACG, from Dictyostelium. The proposed structure for ACA resembles that proposed for mammalian adenylyl cyclases: two large hydrophilic domains and two sets of six transmembrane spans. ACG has a novel structure, reminiscent of the membrane-bound guanylyl cyclases. An aca- mutant, created by gene disruption, has little detectable adenylyl cyclase activity and fails to aggregate, demonstrating that cAMP is required for cell-cell communication. cAMP is not required for motility, chemotaxis, growth, and cell division, which are unaffected. Constitutive expression in aca- cells of either ACA or ACG, which is normally expressed only during germination, restores aggregation and the ability to complete the developmental program. ACA expression restores receptor and guanine nucleotide-regulated adenylyl cyclase activity, while activity in cells expressing ACG is insensitive to these regulators. Although they lack ACA, which has a transporter-like structure, the cells expressing ACG secrete cAMP constitutively.


Proliferating cell nuclear antigen is required for DNA excision repair. Fractionation of extracts from human cell lines allows nucleotide excision repair of damaged DNA to be resolved into discrete incision and polymerization stages. Generation of incised intermediates depends on the XP-A protein, a polypeptide that recognizes sites of damaged DNA, and on the human single-stranded DNA-binding protein HSSB. The proliferating cell nuclear antigen (PCNA) is required for the DNA synthesis that converts the nicked intermediates to completed repair events. This need for PCNA implies that repair synthesis is carried out by DNA polymerase delta or epsilon. The ability to visualize repair intermediates in the absence of PCNA facilitates dissection of the multiprotein reaction that leads to incision of damaged DNA in a major pathway of cellular defense against mutagens.


New approaches to poliovirus diagnosis using laboratory techniques: memorandum from a WHO meeting. Laboratory diagnosis of poliomyelitis is an important part of the WHO initiative for global eradication of poliomyelitis. During the last year, new methods have been developed for the detection of poliovirus in clinical specimens, for intratypic differentiation, for the analysis of poliovirus neurovirulence, and for the detection of poliovirus antibodies. Progress in laboratory techniques for detection of poliovirus antibodies and for characterization of poliovirus isolates has suggested several new approaches to poliovirus diagnosis using laboratory techniques.


Megestrol acetate reverses multidrug resistance and interacts with P-glycoprotein. We evaluated the multidrug resistance (MDR)-modulating effects of progesterone (PRG) and an orally active, structurally related compound, megestrol acetate (MA), in several MDR human cell lines. At 100 microM, both steroids inhibited the binding of a Vinca alkaloid photoaffinity analog to P-glycoprotein (P-gp) in MDR human neuroblastic SH-SY5Y/VCR cells [which show greater than 1500-fold resistance to vincristine (VCR) in the tetrazolium dye (MTT) assay]. However, 100 microM MA markedly enhanced the binding of [3H]-azidopine to P-gp in both SH-SY5Y/VCR cells and the MDR human epidermoid KB-GSV2 cell line (which displays 250-fold resistance to VCR in the MTT assay). PRG had little effect on the binding of [3H]-azidopine to P-gp. MA at low doses was more effective than PRG in sensitizing cells to VCR and enhancing their accumulation of [3H]-VCR. The highly resistant SH-SY5Y/VCR subline exhibited significant collateral sensitivity to both steroids. These data suggest that MA may be a clinically useful modulator of MDR.


Intrinsic resistance to anticancer agents in the murine pancreatic adenocarcinoma PANC02. PANC02 is a ductal adenocarcinoma of the pancreas that is resistant to every known class of clinically active antitumor agent. To study the mechanism(s) underlying the intrinsic drug resistance of this tumor, a mammary adenocarcinoma (CA-755) that also grows in C57/BL mice and is known to be drug sensitive was used for comparison. PANC02 resistance and CA-755 sensitivity to several antitumor agents and to X-ray therapy was confirmed in mice, and PANC02 also demonstrated relative resistance in tissue culture. Relative to Chinese hamster ovary (CHO) and CA-755 cells, PANC02 did not appear to show a higher rate of mutation to drug resistance in culture as based on the 6-thioguanine resistance marker. Although P-glycoprotein characteristic of the multidrug resistance (MDR) phenomenon could be demonstrated at the mRNA level using a sensitive RNAse protection assay, the level of expression found was several orders of magnitude lower than that observed in phenotypic MDR cell lines. Furthermore, quinidine failed to increase the sensitivity of PANC02 cells to Adriamycin under conditions that clearly potentiated the toxicity of the drug to a CHO cell line exhibiting classic MDR traits. The heterogeneity in the distribution of drugs was inferred as being significantly greater in PANC02 versus CA-755 cells in vivo as based on measurements of within-animal, within-tumor variance in the distribution of the marker compounds inulin and antipyrine. Although it may not be the only mechanism involved, this greater intratumor heterogeneity in drug distribution could theoretically play a major role in the intrinsic drug resistance of PANC02 in vivo.


Ca2+ regulation of mechanical properties of striated muscle. Mechanistic studies using extraction and replacement of regulatory proteins. Extraction of regulatory proteins from thick and thin filaments of vertebrate striated muscle has proven to be an important approach in elucidating roles of these proteins in regulating contraction and in probing specific mechanisms of activation. For some proteins, such as LC2 and C protein, extraction has been fundamental in demonstrating the importance of these proteins in modulating contraction and the kinetics of cross-bridge interaction. For other proteins, such as TnC and troponin, extraction has provided significant insight into the importance of thin-filament intermolecular cooperativity in modulating Ca2+ sensitivity of the contractile process. A combination of extraction and readdition has provided a means of introducing mutated or derivatized proteins into fibers to accomplish a variety of experimental objectives. The use of this approach is likely to grow with the need to test the functional consequences of site-specific mutations as part of studies directed to mechanisms of regulation or altered regulation in heart and skeletal muscles under normal and pathophysiological conditions. Such studies are likely to include extraction in combination with other probes of function such as flash photolysis of reaction substrates or products within the cross-bridge interaction cycle. Although extraction is a powerful approach and is likely to be extended to proteins not discussed in this review, an essential element of experimental design in studies such as these is that appropriate control experiments be done to verify that observed effects of the extraction protocol are specifically attributable to the protein that is removed.


Effects of beta-adrenergic receptor stimulation and blockade on rate-dependent atrioventricular nodal properties. Recent work has shown that alterations in the dynamic atrioventricular (AV) nodal response to changes in heart rate can significantly modify AV nodal function. The present study was designed to evaluate the nature and potential importance of sympathetic regulation of the rate-dependent properties of the AV node. Selective stimulation protocols and mathematical formulations were used to independently quantify AV nodal recovery, facilitation, and fatigue in 12 morphine-chloralose-anesthetized dogs. Vagal effects were prevented by bilateral vagal transection and intravenous atropine, and the sinus node was crushed to allow a broader range of pacing cycle lengths. In seven dogs with sympathetic nerves intact, beta-adrenergic receptor blockade increased the recovery time constant (tau rec) for the conduction of premature test beats from 47 +/- 2 (mean +/- SEM) msec (control) to 62 +/- 1 msec (p less than 0.001), whereas isoproterenol decreased tau rec to 38 +/- 1 msec (p less than 0.001). In addition, beta-blockade increased the maximum amount of rate-dependent AV nodal fatigue from 7 +/- 1 msec (at a cycle length of 198 +/- 9 msec [control]) to 17 +/- 2 msec (p less than 0.001). In five dogs with decentralized stellate ganglia, tau rec was decreased from 71 +/- 3 msec (control) to 57 +/- 4 msec and 48 +/- 2 msec (p less than 0.001 for each) by left stellate ganglion stimulation at 5 and 10 Hz, respectively. Maximum fatigue was similarly reduced from 16 +/- 1 msec (control) to 12 +/- 2 msec (p = NS) and 8 +/- 1 msec (p less than 0.01), respectively. Stellate ganglion stimulation, isoproterenol, and beta-blockade did not alter AV nodal facilitation. A mathematical model incorporating quantitative indexes of AV nodal function accurately accounted for tachycardia-dependent increases in the atrial-His activation interval, which were enhanced by beta-adrenergic receptor blockade and reduced by isoproterenol. Furthermore, this model showed that beta-adrenergic effects were increased by increasing heart rate, with the majority of the rate-dependent action being due to changes in the time course of AV nodal recovery. We conclude that beta-adrenergic receptor stimulation alters functional properties that govern the AV nodal response to changes in heart rate. These changes in functional properties alter the ability of the AV node to conduct impulses during tachycardia and, as such, could play a major role in the ability of sympathetic stimulation to promote and beta-adrenergic receptor blockade to prevent the occurrence of AV nodal reentrant arrhythmias.


Pituitary imaging using a labelled somatostatin analogue in acromegaly. A number of neoplasms are known to express somatostatin receptors, and the use of somatostatin receptor imaging in their localization has recently been described. We have looked at the use of an 123I-labelled Tyr3-octreotide analogue of somatostatin in the visualization and functional characterization of growth hormone-secreting pituitary adenomas. Fifteen patients with biochemically-proven acromegaly were scanned using this agent. In eight of these we also assessed acute GH responses to octreotide in order to correlate these responses with tumour uptake characteristics. Planar and single-photon-emission computerized tomographic (SPECT) images of the head were obtained using a gamma-camera at 10 minutes, and 4 and 24 hours, after injection of the radiopharmaceutical. Blood for serum GH was sampled for 12 hours after administration of a single dose of 100 micrograms octreotide. Twelve of the acromegalic subjects showed significant uptake of the radiopharmaceutical in the pituitary fossa. Of the eight patients in whom we assessed acute GH responses, five demonstrated a significant fall in GH in response to octreotide. These subjects also showed positive uptake in the pituitary on scanning. The three patients who had no fall in GH had no uptake on scanning. Uptake of 123I-labelled Tyr3-octreotide in the pituitary fossa appears to correlate closely with the presence of a therapeutic response to octreotide.


Long-term treatment with the dopamine agonist CV 205-502 of patients with a clinically non-functioning, gonadotroph, or alpha-subunit secreting pituitary adenoma. We aimed to assess the effects of prolonged treatment with the dopamine agonist CV 205-502 on tumour volume, visual field defects, and serum gonadotrophin and alpha-subunit concentrations in patients with gonadotroph, alpha-subunit secreting, or clinically non-functioning pituitary adenomas. The patients were treated with CV 205-502 in a final daily dose of 300 micrograms for at least 1 year. The patients were seen at 2 or 3-week intervals during the first 3 months of treatment, and thereafter every 1 or 2 months. Computerized tomography and Goldmann perimetry were performed before treatment and during follow-up. Blood samples were drawn before treatment and at each out-patient visit. One patient with gonadotroph, two with alpha-subunit secreting, and two with clinically non-functioning pituitary adenomas were studied. Computerized tomography showed tumour shrinkage in one patient. In two other patients an improvement of visual field defects was observed. In four patients, a significant decrease in serum FSH and/or alpha-subunit concentrations occurred within the first 3 months of treatment. In the remaining patient, a significant decrease of serum FSH and alpha-subunit concentrations was found after more than 3 months of treatment. In patients with clinically non-functioning, gonadotroph, or alpha-subunit secreting pituitary tumours, long-term treatment with the dopamine agonist CV 205-502 decreases serum FSH and/or alpha-subunit concentrations. This decreased secretory activity from the pituitary tumour may be accompanied by an improvement of visual field defects, or tumour shrinkage on computerized tomography. Therefore, treatment with CV 205-502 may be useful in patients with clinically non-functioning, gonadotroph, or alpha-subunit secreting pituitary tumours, who cannot be operated upon.


Distribution and quantification of somatostatin in inflammatory disease. To study the possible alteration of mucosal-submucosal somatostatin-containing cells in inflammatory bowel diseases (IBD), the total numbers of somatostatin-containing endocrine cells (SCEC) and submucosal ganglion cells (SGC) were counted in Crohn's disease (CD) and ulcerative colitis (UC). Tissue specimens from 25 CD and 25 UC patients were fixed in Hollande's fixative immediately after resection and were investigated by immunohistochemical staining. A single specimen was collected from 25 colorectal cancer patients, the control group. There was a significant difference in the number of SCEC between the tissues taken from the proximal colon (ascending and transverse colon) and the distal colon (descending and sigmoid colon). The distal colon tended to contain more somatostatin-immunoreactive cells than did the proximal colon. In IBD, SCEC were decreased in number compared with the controls. This decrease was related to the degree of inflammation in CD; the higher the grade of inflammation, the lower the number of SCEC. The number of SGC was decreased in IBD: however, a significant decrease was noticed only in CD. The anatomic origin and the degree of inflammation did not affect the number of SGC. In the present study, the decrease of somatostatin-containing cells was noticed in both CD and UC, but there was no significant difference between CD and UC. Therefore, it was assumed that this decrease was secondary to inflammation. However, the decrease of somatostatin, which works as an inhibitory peptide for inflammation, might have some role in the pathogenesis of IBD.


Effect of erythropoietic stress on donor hematopoietic cell expression in chimeric rhesus monkeys transplanted in utero. We have previously reported the successful development of hematopoietic chimerism after the in utero transplantation of fetal hematopoietic stem cells (HSC) in rhesus monkeys (Macaca mulatta). These animals exhibit sustained engraftment without immunosuppression or graft-versus-host disease (GVHD). To assess the functional response of the donor-derived erythropoietic population, we assayed the relative expression of donor and recipient hematopoietic progenitors in chimeric monkeys before and after anemic stress. Anemia in our chimeric animals resulted in increased erythropoietin (EPO) production comparable to controls. This was accompanied by changes in erythroid progenitor profiles, again similar to controls. Chimeric animals demonstrated normal reticulocytosis and reconstituted their hematocrit after hemorrhage at the same rate as controls. The donor-derived erythropoietic population exhibited normal responses to recipient regulatory signals and did not seem to expand at the expense of other hematopoietic lineages. Thus the proportions of engraftment for the myeloid and erythroid precursors in bone marrow and for blood lymphocytes remained stable. Our results demonstrate that the in utero transplantation of fetal HSC results in stable engraftment of donor erythropoietic progenitors, which appear to be fully integrated within the recipient's regulatory system. The abnormalities reported in the postnatal transplantation setting can then be attributed to immunologic reactions requiring conditioning myeloablative regimens. Fetal transplantation bypasses all these factors.


Heterotypic adherence between murine leukemia/lymphoma cells and marrow stromal cells involves a recognition mechanism with galactosyl and mannosyl specificities. We have previously shown that leukemia/lymphoma (LL) cells adhere to marrow stromal cells (MSC), and MSC induce clonal growth of LL cells. Even though CD11a/18 and CD54 are important in leukocyte and endothelial cell interaction, the literature suggested that these adhesion proteins are not involved in adhesion of hematopoietic stem cells to MSC. We therefore utilized a unique ICAM-1- murine MSC line (MLT) to evaluate the mechanisms of adherence of LL cells (L5178Y and L1210) to MLT. Adherence of LL cells to extracellular matrix (ECM) proteins was also examined. L1210 cells attached to collagen types III and IV, laminin, and fibronectin, but not to collagen type I. L5178Y cells did not attach to any of the ECM proteins tested. The adherence of both L1210 and L5178Y to MSC was unaffected by rat monoclonal antibodies to murine CD11a, CD11b, and CD18. Neoglycoprotein probes, mannosyl-bovine serum albumin (BSA) and galactosyl-BSA, inhibited the adherence of L5178Y and L1210 cells to MSC by 34%-63% of controls at concentrations of 10(-3) and 5 x 10(-3) M. In contrast, fucosyl-BSA had no inhibitory effect on LL cell adherence of MLT. These data suggest that 1) LL cells may adhere to MSC by a lectin mechanism with mannosyl and galactosyl specificities; and 2) other mechanisms of adherence, not yet defined, are also important in this system.


Synergism in cytosolic Ca2+ mobilization between bradykinin and agonists for pertussis toxin-sensitive G-protein-coupled receptors in NG 108-15 cells. Bradykinin (BK) induced a transient and pertussis toxin (PT)-insensitive increase in cytosolic Ca2+ ([Ca2+]i) in NG 108-15 neuroblastoma x glioma hybrid cells, whereas leucine-enkephalin (EK), somatostatin, norepinephrine or carbachol showed a weak but PT-sensitive action. When any one of the latter agonists was applied to the cells treated with low doses of BK, however, the level of [Ca2+]i rise caused by the agonist was remarkably increased in a PT-sensitive manner. The decreasing of extracellular Ca2+ only slightly influenced the actions of these agonists. Thus, synergism between a BK receptor and PT-sensitive G-protein-coupled receptors results in marked intracellular Ca2+ mobilization by the latter agonists.


Adjunctive antibiotic treatment of women with preterm rupture of membranes or preterm labor. Subclinical infection is associated with preterm rupture of the membranes (PROM) and preterm labor (PTL) in many cases. It was hypothesized that antibiotic treatment might delay delivery and/or decrease infectious morbidity in those with PROM or PTL. Patients from 19 to 34 weeks with PROM and no labor or PTL with intact membranes (but not both) were separately randomized to receive ampicillin versus placebo in addition to usual therapy. There were 36 women with PTL (21 ampicillin/15 placebo) and 84 with preterm PROM (41 ampicillin/43 placebo). Demographically, the treatment and placebo groups were similar. Outcome variables analyzed included delivery delay after treatment, maternal chorioamnionitis/endometritis, Apgar score, neonatal infection, or respiratory distress, and hospital stay. There were no significant differences between the ampicillin and placebo groups in those with PTL or preterm PROM as it concerned outcome parameters. Adjunctive ampicillin used for treatment of idiopathic PTL or preterm PROM was not beneficial in this study.


Presentation and management of eclampsia. Three hundred forty-seven cases of eclampsia were managed at the University College Hospital (UCH), Ibadan, Nigeria from 1977 to 1986 (9.3 per 1000 deliveries). Thirty-one percent of seizures occurred antenatally, 46.2% during labor and 23.2% postnatally. Only 5% first occurred in the hospital. Seizures were controlled with diazepam, lytic cocktail (chlorpromazine, pethidine and phenergan), sodium amylobarbitone, paraldehyde and bromethol. Maternal mortality was 2.9%, and perinatal mortality 193 per 1000, respectively. Prevention of avoidable factors (absent or poor antenatal care and prolonged labor) by the provision of comprehensive antenatal care will reduce the incidence of eclampsia. Improvement in management facilities prior to transfer to referral centers, the use of magnesium sulfate or diazepam to control seizures and the avoidance of hypotonic solutions and 50% glucose during therapy will reduce morbidity and mortality.


[The characteristics of the effect of mediator substances on the dynamic oxygen tension in the gastric mucosa]. The studies on rats were carried out to determine dynamics of pO2 in the mucous membrane of the stomach under the effect of acetylcholine, norepinephrine, serotonin, histamine, prostaglandin E2 and ATP. As to the changes in pO2 the mediator substances were arranged as follows (from more intensive effect to less pronounced one): serotonin, acetylcholine, prostaglandin E2, norepinephrine, histamine and ATP. As to the duration of the action--PGE2 acetylcholine, serotonin, norepinephrine, ATP and histamine. Under the joint action of the mediator substances (serotonin, norepinephrine, histamine) with acetylcholine the effects of domination and modulation of acetylcholine effect are found.


Anticardiolipin antibodies in eclampsia. Twenty patients with eclampsia were studied for the presence of anticardiolipin antibodies by the standard microflocculation test (quantitative) used in the VDRL. Four out of 20 patients (20%) were positive for anticardiolipin antibodies. Anticardiolipin antibodies which are antiphospholipid antibodies by decreasing the prostacyclin production may damage the vascular endothelium and cause thrombosis in the vessels. Hence, the presence of anticardiolipin antibody in eclampsia may have association with antiphospholipid antibody syndrome described by Harris et al. (Lancet ii: 1211, 1983) and its central nervous system sequelae.


Amoxycillin/clavulanic acid (augmentin) compared with triple drug therapy for pelvic inflammatory disease. Sixty Jordanian women with pelvic inflammatory disease (PID) were studied. Of these, 31 were given oral amoxycillin/clavulanic acid (augmentin) for a mean duration of 8.4 days and 29 were given a standard triple drug regimen of oral ampicillin, intramuscular gentamicin and metronidazole tablets/pessaries for a mean duration of 7.2 days. Bacterial culture (cervical and high vaginal swabs) was positive in every case, most often E. coli but sometimes more than one pathogen was isolated. No gonococci were isolated and tests for chlamydia in 16 patients (8 in each group) were negative, suggesting a dissociation between the etiology of PID and sexually transmitted disease in this Jordanian study. After 3 days of treatment, more patients in group I (augmentin) showed diminution of symptoms of pain and discharge (P less than or equal to 0.05) compared to group II. At the end of treatment, complete cure or satisfactory improvement was recorded in 93.1% and 92.9% of cases in the two groups, corresponding to in vitro bacterial efficacy of 90.4% and 96.5%, respectively. No serious side effects were noted in either regimen. The results of this comparative study suggest that oral amoxycillin/clavulanic acid (augmentin) may be a convenient alternative to the triple drug regimen usually administered for the treatment of pelvic inflammatory disease.


Strangulation of the umbilical cord due to combined amniotic band and true knot. A case of fetal death due to an amniotic band and an umbilical cord true knot is presented. To the best of our knowledge the simultaneous occurrence of true knot of cord and amniotic band has never been previously reported and may shed light on the possible time of true knot formation in this case.


Cervical priming and labor induction by multiple doses of intracervical prostaglandin E2 gel. One hundred seventy-two term pregnant women with medical or obstetric conditions requiring induction of labor were treated with intracervical administration of 0.5 mg prostaglandin E2 in tylose gel. Multiple administrations were necessary in 42 cases (24.4%), two administrations in 31 cases (18.0%) and three administrations in 11 cases (6.4%). Intracervical administration of PGE2 tylose gel (0.5 mg dose) is useful to prime the cervix, induce labor, and significantly modify Bishop score.


Operative vaginal delivery. ACOG Technical Bulletin. Number 152--February 1991. With any operative vaginal delivery, it is important to document in the medical record the indications for the procedure, including the position and station of the vertex and specifics of technique, timing, and ease of the procedure. The use of cord pH and blood gas determinations may be helpful for further evaluation of the fetus.


Plasminogen activators and inhibitors in normal late pregnancy, postpartum and in the postnatal period. Tissue plasminogen activator (t-PA), urokinase plasminogen activator (u-PA) and plasminogen activator inhibitors (PAI) are elevated in late pregnancy with t-PA and u-PA remaining so at 6 weeks postnatal. PAI-2 remains at postpartum but was absent by 6 weeks postnatal unlike PAI activity which was absent at postpartum and returned to nonpregnant level at postnatal. The potential fibrinolytic response to stress is much reduced in pregnancy thus increasing the risk of thromboembolism.


Linkage analysis of insulin-receptor gene in familial NIDDM. Although non-insulin-dependent diabetes mellitus (NIDDM) is clearly inherited, the mode of inheritance and genetic etiology remain unknown. Impaired insulin action is an important component of NIDDM, which may precede NIDDM onset, and appears to be inherited. Numerous defects of the insulin-receptor gene have been described in syndromes of extreme insulin resistance, and this gene is a strong candidate for genetic predisposition to NIDDM. To test this hypothesis, we examined 18 white pedigrees from Utah that had two or more siblings with NIDDM. For each pedigree, individuals not known to be affected were tested by standard oral glucose tolerance test, and diagnoses of NIDDM and impaired glucose tolerance were made by World Health Organization criteria. Each individual was typed for seven restriction-fragment-length polymorphism markers at the insulin-receptor locus, and marker phase was established by segregation. Linkage was examined with the LINKAGE program under six models, including autosomal dominant and autosomal recessive, with individuals with impaired glucose tolerance counted either as affected or of unknown status and with or without sporadic cases of diabetes. Under each model, linkage was significantly rejected. Neither inspection of individual pedigree log of odds scores nor formal tests of heterogeneity suggested a subgroup in which linkage of NIDDM and insulin-receptor gene was likely. In addition, sharing of insulin-receptor gene haplotypes among 108 affected sibling pairs drawn from the pedigrees did not deviate from that expected by chance alone.(ABSTRACT TRUNCATED AT 250 WORDS)


Management of massive upper gastrointestinal haemorrhage from multiple sites of peptic ulceration with somatostatin and octreotide--a report of five cases. Surgical management of massive upper gastrointestinal bleeding after failed medical treatment may be hazardous because of diffuse bleeding from several sites, further complicated in some patients by intercurrent disease, age, or previous surgery. Experience with combined somatostatin and octreotide therapy in five such patients is described. All were treated initially with either intravenous somatostatin (250 micrograms/hour) or octreotide (Sandostatin) (50 micrograms/hour) for periods ranging from three to five days, after which they were given subcutaneous octreotide (50 or 100 micrograms three times daily). Bleeding was controlled by this regimen in all cases. The patients were all discharged from hospital on either ranitidine (n = 4) or omeprazole (n = 1). Repeat endoscopy at the end of the treatment period with somatostatin and octreotide (n = 1) or four weeks after discharge (n = 3) showed complete healing of the bleeding sites. Somatostatin and octreotide may be of value in controlling severe upper gastrointestinal bleeding in patients in whom surgery is hazardous because of bleeding from several peptic lesions further complicated in some by intercurrent disease or age.


Enteral nutrition in malnutrition following gastric resection and cephalic pancreaticoduodenectomy. Nutritional recovery was studied during continuous enteral nutrition in 29 patients who had developed malnutrition after gastric surgery. Patients were divided into three groups according to the type of surgery involved: total gastrectomy (n = 10), partial gastrectomy (n = 12), or cephalic pancreaticoduodenectomy (n = 7). The evolution of anthropometric and biological nutritional parameters in each group was compared with that observed in a control group of 10 nonoperated anorectic patients. Significant gains in body weight, arm muscle circumference, triceps skinfold, serum transferrin and global nutritional status were observed after 3 to 4 weeks of enteral nutrition in each group, while serum albumin, serum cholesterol, hemoglobin, and total lymphocyte count did not change significantly. No significant difference was observed between the groups. However, weight gain tended to be slower in patients with cephalic pancreaticoduodenectomy. This study confirms that enteral nutrition is an effective method of nutritional repletion after gastrectomy. Enteral nutrition can be used in undernourished gastrectomized patients when dietary measures alone have proven inadequate.


Genetic divergence between the Wistar-Kyoto rat and the spontaneously hypertensive rat. A method of restriction fragment length polymorphism (RFLP) analysis was used to estimate the amount of genetic divergence between the spontaneously hypertensive rat (SHR) strain and the Wistar-Kyoto (WKY) strain. DNA from each strain was digested with eight restriction endonucleases and hybridized with six single copy gene sequences. The number of hybridization bands in each digestion was used to estimate the total number of bases analyzed and RFLPs were scored as single mutations. Divergence was then estimated by dividing the number of mutations by the number of bases analyzed. In a total of 808 bases analyzed in WKY rats, a minimum of 13 mutations were scored in SHR, which yields a nucleotide divergence of 1 change per 62 bp. This is an extremely high amount of divergence given the known origin of these two strains and is comparable to the maximum divergence possible between unrelated humans.


Treatment of acute variceal bleeding.

Treatment of acute variceal bleeding. Once the bleeding patient has been resuscitated and the diagnosis of acute variceal hemorrhage established by endoscopy, emergency injection sclerotherapy should be employed as the therapeutic option of choice. Endoscopic band ligation is a promising new technique that may prove to be as effective as sclerotherapy, with fewer complications. Pharmacologic treatment (with vasopressin and nitroglycerin) and balloon tamponade remain important alternative treatments, both as empiric temporizing therapy before sclerotherapy can be arranged and in the approximately 30% of patients who continue to bleed after a single sclerotherapy session. Continued bleeding in many of these patients can be controlled with a second session of sclerotherapy. If active acute bleeding persists after two sclerotherapy treatments, treatment should be considered a failure. Some of these patients may be suitable for surgical treatment with either staple-gun transection of the esophagus or emergency portacaval shunting.


Prevention of variceal rebleeding.

Prevention of variceal rebleeding. Recurrent variceal hemorrhage occurs in 50% to 80% of cirrhotic patients who survive a variceal bleeding episode. The aim of preventing rebleeding is to improve survival by reducing the mortality associated with rebleeding; however, although shunt surgery is the most effective treatment to prevent recurrent bleeding, it does not increase survival and is associated with an increased incidence of chronic portosystemic encephalopathy. The distal splenorenal shunt is associated with a reduced incidence of encephalopathy, compared with nonselective shunts, but the true magnitude and longevity of this effect is still controversial. beta-Blockers reduce the incidence of rebleeding, but the effect is modest, and there is little or no effect on mortality when compared with no treatment. Injection sclerotherapy reduces the incidence of rebleeding and improves survival when a schedule of both emergency and long-term injection is compared with no sclerotherapy. No technical variation of injection sclerotherapy has been shown to be superior to another. Endoscopic variceal banding may result in fewer complications but the efficacy is similar to that of injection sclerotherapy. Trials of long-term sclerotherapy versus beta-blockers show very similar mortality and rebleeding rates. Addition of beta-blockade to sclerotherapy does not confer any advantages when compared with sclerotherapy alone. Improvements in pharmacologic therapy, such as the addition of isosorbide mononitrate to propranolol, may in the future make drug therapy the first treatment option to prevent rebleeding. Shunt surgery is superior to sclerotherapy in preventing rebleeding and has a similar mortality; however, liver transplantation is technically more difficult in shunted patients, but shunts do not adversely affect overall survival after transplantation. There are few data to allow optimal selection of a particular therapy or sequence of therapies to prevent variceal rebleeding for any individual patient. This will need to be studied in large trials and is a major issue in the current clinical management of cirrhotics who have bled from varices.


Prevention of initial variceal hemorrhage. Results from prospective controlled trials do not justify the use of either prophylactic shunt surgery or sclerotherapy for the prevention of initial variceal bleeding. Use of nonselective beta-adrenergic blockers has been shown to reduce significantly the risk of first variceal hemorrhage, but their effect on survival is marginal. Because only 25% to 40% of patients with cirrhosis and varices experience variceal bleeding, is it justified to place all patients with varices on beta-blocker therapy? The answer lies in the identification of a high-risk population (i.e., patients with large varices and endoscopic red color signs) and patients who can tolerate and will be compliant with therapy. The duration of therapy required is unknown and perhaps requires a lifetime commitment. Future research will involve the use of combinations of pharmacologic agents to reduce further portal pressure and perhaps the use of pharmacologic agents and sclerotherapy. The goal of a significant improvement in survival is yet to be obtained.


Pharmacologic treatment of portal hypertension. Variceal formation and rupture are dreaded complications of chronic liver disease and portal hypertension. The pharmacologic treatment of portal hypertension should be able to stop as well as to prevent variceal hemorrhage. There are two principal types of vasoactive drugs in the treatment of portal hypertension: vasoconstrictors and vasodilators. Vasoconstrictors reduce the splanchnic blood flow, thereby decreasing the portal blood flow and portal pressure. Vasodilators act by different mechanisms, including by relaxation of myofibroblasts in the fibrous septa and presinusoidal areas of the liver and by direct vasodilation of the collateral circulation. In addition, paradoxically, they could decrease portal flow and pressure by inducing a baroreflex-mediated mesenteric arterial vasoconstriction. A miscellaneous group of drugs is also available. These drugs reduce the blood flow and pressure in the gastroesophageal variceal system by mechanisms other than vasoconstriction or vasodilation. The success of these pharmacologic agents is limited once the varices have ruptured. The use of beta-blockers in the prophylaxis of the first variceal bleeding has been proven of benefit in this respect. Future research should be aimed at elucidating the role that humoral and endothelial factors play in development of the hyperdynamic circulatory state that characterizes patients with portal hypertension. Once these etiologic factors have been identified and new knowledge is acquired about their role in the complications of chronic liver disease, the challenge will rest on developing novel pharmacologic therapies specifically targeting these factors.