Dopamine receptor sequences. Therapeutic levels of neuroleptics occupy D2 receptors, clozapine occupies D4. Dopamine (DA) D2, D3, and D4 receptors are targets for antipsychotic drugs. The recent cloning, deoxyribonucleic acid sequencing, and brain location of these receptors provide new insight on the DA hypothesis of schizophrenia, particularly for the basis of antipsychotic therapy of schizophrenia. In schizophrenia brain tissue, D2 receptors are elevated and have lost the link to D1 receptors. Brain positron-emission tomography studies of patients may also reveal elevated D2, depending on the method used. Hallucinations and positive symptoms are blocked when about 70% of the D2 receptors are occupied by neuroleptic drugs. An analysis of the literature indicates that therapeutic concentrations of antipsychotic drugs (in the patient's cerebrospinal fluid or plasma water) act primarily at D2 receptors, with the exception of clozapine, which acts at D4 receptors.


Processed MHC class I alloantigen as the stimulus for CD4+ T-cell dependent antibody-mediated graft rejection. The traditional view of graft rejection is one of direct recognition of allogeneic MHC molecules by effector T cells, the phenotype of which may be predicted by the nature of the MHC disparity. In this article, Andrew Bradley and colleagues discuss recent evidence that suggests this view may be an oversimplification, and argue that additional effector mechanisms, such as alloantibody, need to be reconsidered.


An association between a Bc1I restriction fragment length polymorphism of the glucocorticoid receptor locus and hyperinsulinaemia in obese women. Obesity is likely to be a multifactorial disease with an important genetic component. Animal models of genetic and experimentally induced obesity suggest that glucocorticoid receptor (GR) activity plays a role in the aetiology and maintenance of the obese state. Glucocorticoid activity appears to be essential for the development of hyperinsulinaemia and subsequent fat deposition. In humans, glucocorticoid excess is associated with central fat distribution. We have therefore investigated the restriction fragment length polymorphisms of the human GR gene locus (GRL) and have sought associations of specific alleles with anthropometric measurements and indices of insulin secretion and resistance in obesity. Fifty-six extremely obese, unrelated, nondiabetic premenopausal British Caucasian females and 43 age-matched, normal weight controls were studied. The obese subjects were characterized by fat distribution (waist to hip ratio), insulin secretion and insulin resistance (fasting insulin (FI)), an index of insulin resistance (HOMA), stimulated insulin secretion during an oral glucose tolerance test and insulin-mediated glucose disposal, steady-state plasma glucose). A Bc1I polymorphism (fragments of 4.5 and 2.3 kb) demonstrated significant association with indices of glucose metabolism in obesity; those subjects homozygous for the 4.5 kb fragment had elevated FI (Pc = 0.012) and HOMA (Pc = 0.012) values. The genotypic and allelic frequencies of the GRL Bc1I polymorphism were otherwise similar in obese and normal weight subjects. We postulate that the GRL Bc1I polymorphism may directly affect GR gene expression, or be in linkage disequilibrium with a possible mutation within one of three exons of the GR gene, and thereby modulate GR transcriptional activity on target genes involved in glucose and insulin homeostasis.


The role of cytokines in tumor immunotherapy. Report on the 2nd Frankfurt International Cytokine Symposium 25-27 June 1992, Frankfurter Hof, Frankfurt, Germany. The conference, organized by Profs. Mitrou, Bergmann (Frankfurt), Huber (Mainz) and Niederle (Leverkusen), concentrated almost exclusively on the role of cytokines in cancer. The majority of presentations concerned IFN-alpha, IL 2 or TNF-alpha, but G-CSF, GM-CSF, IL 4, IL 10 and TGF-beta were not neglected. Presentations achieved a laudable balance between basic science and clinically oriented studies. The present report emphasizes the clinical aspects; proceedings of the entire meeting will be published by S. Karger AG, Basel.


Development of tyrosine hydroxylase-, dopamine- and dopamine beta-hydroxylase-immunoreactive neurons in a teleost, the three-spined stickleback. The development of catecholaminergic neuronal systems in the brain of a teleost, the three-spined stickleback, was studied through embryonic to early larval stages by immunocytochemistry using specific antibodies against dopamine, tyrosine hydroxylase and dopamine beta-hydroxylase. By analysing the spatiotemporal patterns of development for the catecholaminergic nuclei, possible homologies with nuclei in amniote brains have been identified. The noradrenergic neurons in the isthmus region of the rostral rhombencephalon originate in the same manner as the A4-A7 + subcoeruleus group in mammals. Their developmental characteristics show the largest similarities with the subcoeruleus group of birds and mammals, although some features are shared with developing A6 (locus coeruleus) neurons. Catecholaminergic neurons never appear during development in the ventral mesencephalon of the three-spined stickleback. A group of large dopaminergic neurons that accompany the cerebrospinal fluid (CSF)-contacting neurons follows the border between the hypothalamus and the ventral thalamus into the caudal hypothalamus, where they are continuous with the dopaminergic neurons in the posterior tuberculum. They are thus topologically comparable with the dopaminergic neurons of the zona incerta in mammals. The dopaminergic CSF-contacting neurons that line the median, lateral and posterior recesses of the third ventricle do not contain tyrosine hydroxylase-immunoreactivity at any developmental stage. This indicates that they take up and accumulate exogenous dopamine or L-dihydroxyphenylalanine, and do not synthesize dopamine from tyrosine at any developmental stage. Tyrosine hydroxylase-immunoreactive neurons appear in the pineal organ on the day of hatching (120 h post-fertilization). They were still observed in 240-h-old larvae, but are absent in the pineal organ of adult sticklebacks. The initial appearance and subsequent differentiation of catecholaminergic neurons in the stickleback embryo follow essentially the same spatial and temporal pattern as in amphibian, avian and mammalian embryos. This observation supports the hypothesis that morphologically, topologically and chemically similar monoaminergic neurons in different vertebrate classes are homologous.


Effect of methylation inhibitors on gene expression in HL-60 cells. The methylation inhibitors Neplanocin A (Nep A), 3'-deazaadenosine (dzAdo), and 3-deaza(+/-)aristeromycin (Dari) were tested for their effect on the expression of histone H2B, actin, and the protooncogenes c-myc, and v-fos. Nep A and Dari bind to the S-adenosylhomocysteine hydrolase resulting in the accumulation of S-adenosylhomocysteine, while dzAdo served as a substrate for the enzyme. With dzAdo, inordinant amounts of 3-deazaadenosylhomocysteine (dzAdoHcy) accumulated in the cell, provided L-homocysteine (Hcy) was present. When added at sublethal concentrations, the methylation inhibitors had little or no effect on c-myc, v-fos, histone H2B, or actin expression, nor did any significant number of the drug-treated cells demonstrate myeloid characteristics. However, growth and gene expression were markedly inhibited upon the addition of Hcy and dzAdo. One of the earliest effects of dzAdoHcy on HL-60 cells was the disappearance of c-myc mRNA. Within 1 h of the addition of dzAdo and Hcy, only trace amounts of c-myc mRNA were detectable. After 4-5 h v-fos, histone H2B, and actin mRNAs also decreased to about 40% of control levels. Differences in the stability of preexisting mRNAs would appear to account for these results. Within 1 h following the addition of dzAdo and Hcy, the synthesis of rRNA and mRNA were completely blocked as measured by the incorporation of [3H]uridine.


Distribution of somatostatin-immunoreactivity in the brain of the larval lamprey (Petromyzon marinus). The detailed distribution of somatostatinergic neurons and fibre tracts in the brain of larval lamprey was studied in serially sectioned material using immunocytochemical techniques. Neurons were found to be arranged in four nuclei: a hypothalamic nucleus consisting of both small cerebrospinal fluid-contacting neurons and larger non-contacting neurons, a thalamomesencephalic nucleus and two isthmotrigeminal reticular nuclei. The hypothalamic nucleus is the first to differentiate. Analysis of young larvae showed that somatostatin-immunoreactivity first appeared in hypothalamic cells (12 mm larvae), while it appeared later in the other nuclei. The different somatostatin-immunoreactive fibre tracts innervate different regions of the brain. In addition, somatostatin-immunoreactive fibres originating from hypothalamic neurons were found in the anterior neurohypophysis, which suggests the presence of a hypothalamohypophysial somatostatinergic system in lampreys.


Preparation of plasmid DNA by sequential enzymatic digestion. A new method for the preparation of plasmid DNA from Escherichia coli, sequential enzymatic digestion, is described. The method is based on sequential and selective enzymatic digestion of all components of E. coli except for the supercoiled plasmid DNA. The key enzymes are exonuclease I and exonuclease III that specifically hydrolyze linear chromosomal DNA and are unable to attack supercoiled plasmid DNA under controlled conditions. Isolated plasmid DNA can be sequenced and digested with restriction enzymes.


Evaluation of DNA probe removal from nylon membrane. Genetic fingerprinting is one of the most challenging applications of any hybridization membranes. Forensic DNA fingerprinting typically uses samples in the range of 100-400 ng of genomic DNA. To ensure the ability to successfully reprobe the samples, it is imperative that repeated stripping of sample DNA be minimized while stripping of the probe DNA be maximized. By using standard dilutions of K562 cell line, we compared the following three stripping techniques: NaOH at 25 degrees C, formamide (HCONH2) at 65 degrees C and 0.1 x standard saline citrate and 0.1% sodium dodecyl sulfate at 95 degrees C (high-temperature stripping). The largest amount of genomic DNA was stripped from the membrane with NaOH, with the other two techniques removing less. Formamide and high-temperature procedures resulted in a loss of approximately 5-10 ng of DNA per strip. In contrast, the NaOH resulted in a loss of approximately 10-20 ng per strip.


[Pulmonary artery involvement in Takayasu arteritis. A case of right ventricular failure as presentation form]. Pulmonary involvement in Takayasu's artery disease has been reported since 1940 with an incidence of 14 to 56%. However, the development of severe pulmonary hypertension is an extremely rare event in the natural course of the disease. The authors report a case of a 62 year old male presenting with severe congestive heart failure of recent onset. The initial evaluation and routine exams suggested the presence of pulmonary hypertension of unknown etiology. The absence of left radial pulse in the physical examination led to the performance of a complete angiographic study which confirmed the diagnosis of Takayasu's arteritis with pulmonary involvement and severe pulmonary hypertension.


The activity of dipeptidyl peptidase II and dipeptidyl peptidase IV in mice immunized with type II collagen. We investigated the activity of peptidases in the serum of mice with experimental polyarthritis that was induced by the injection of type II collagen, an experimental model of human rheumatoid arthritis. The activity of dipeptidyl peptidase II (DPP II) was increased and that of dipeptidyl peptidase IV (DPP IV) was decreased resulting in the significant increase of the serum DPP II/DPP IV ratio in the polyarthritic mice compared with that of controls. These results indicate that the DPP II/DPP IV ratio is a novel index of disease activity in mice with collagen-induced polyarthritis and may be useful in assessing the activity of rheumatoid arthritis in humans.


Choice of enzymes for mapping based on CpG islands in the human genome. The frequencies of sites for rare-cutting restriction enzymes in 2.9 million bp of human genomic DNA sequence in the EMBL database have been determined and compared with the expected frequencies. Rare cutters can be divided into four groups based on certain features of their recognition sites. Mlu, I, Nru I, Spl I, and Pvu I are predicted to cleave genomic DNA most infrequently, which is borne out by the fragment lengths observed for Mlu I and Nru I. Thus, these four enzymes are ideal for making long-range maps based on pulsed-field electrophoresis. Other enzymes like Not I are useful for making more detailed maps. Finer maps for identification of CpG islands and associated genes should involve several rare cutters including Eag I, Sac II and Bss HII. A cluster of sites for at least two such enzymes is a good indicator of a CpG island, and 78% of the island-associated genes can be located in this way.


Hox codes and positional specification in vertebrate embryonic axes. We have compared the ways in which vertebrate Hox genes are used in the patterning of three distinct embryonic contexts, the branchial region, the somites, and the limb. We have identified common features of the three systems, but have suggested on the basis of their differences (in both embryological properties and use of Hox genes) that it is better to consider each as an independent system for regional specification. Nevertheless, there are sufficient common features to expect that exploitation of the distinct experimental advantages of each system will provide important insights to the mode of operation of the others.


Effects of cyclosporin A on resident and passenger immune cells of normal human skin and UV-induced erythema reactions. Cyclosporin A (CyA) has proved effective in various dermatological diseases, but its mechanisms of action within the skin are still ill-defined. In order to characterize more clearly the cellular targets of CyA we examined its effects on skin immune cells in normal and UVB- and PUVA-irradiated skin by means of a three-step immunoperoxidase reaction and immunofluorescence double-labelling technique. The CyA-induced depletion of epidermal Langerhans cells equals that seen with UVB or PUVA alone. CyA alone has no effect on the number and distribution of dermal cells. CyA modulates the UV irradiation-induced changes by: (i) inhibiting the UVB- and PUVA-induced ICAM-1 expression by keratinocytes and (ii) suppressing the PUVA-induced upregulation of CD11a expression by macrophages (72 +/- 12% of Ki-M8+ macrophages express CD11a with PUVA, compared with 20 +/- 5% with CyA + PUVA, P < 0.001). Neither treatment affected ICAM-1 expression by endothelial cells. In addition, CyA increases PUVA minimal phototoxicity dose from 10 +/- 2.6 J/cm2 (PUVA alone) to 15.3 +/- 3.1 J/cm2 (CyA + PUVA), (P < 0.001). In conclusion, the effects of CyA on the skin include a down-modulation of the PUVA-erythema reaction, associated with a direct or indirect modulation of adhesion molecule expression.


The preferential expansion of functional CD4+ lymphocyte populations in vitro. We describe a simple and inexpensive chemical procedure for the selective expansion of human CD4+ lymphocytes. The method employs L-leucine methyl ester (LME) to deplete monocytes and large granular lymphocytes, as well as to inhibit growth of CD8+ lymphocytes. LME treatment eliminates granular cells, but most CD8+ lymphocytes, B-lymphocytes, and CD4+ lymphocytes remain. Peripheral blood mononuclear cells (PBMCs) from normal and HIV-positive individuals are treated with LME for 1 h at ambient temperature and cultured in the presence of IL-2 to expand the cell number. Stimulation with the T-cell mitogens concanavalin A, phytohemagglutinin, or OKT3 antibody augments lymphocyte expansion and within 1-3 weeks the culture is greatly enriched (90-100%) in CD4+ lymphocytes. LME-treated lymphocytes expand up to 10-fold during culture in the presence of IL-2 alone and up to 400-fold following treatment with T-cell mitogens. The immune function of LME-treated and expanded peripheral blood lymphocytes was examined using the response to the recall antigens tetanus toxoid and Candida albicans. Fresh PBMCs exposed to these recall antigens proliferated readily. Similarly, LME-treated lymphocytes following expansion responded to these recall antigens with good fidelity to the original PBMC response patterns in four of six donors. The expanded and LME-treated lymphocytes also exhibited good mitogen responses in three of three donors. The LME procedure allows for the simple and inexpensive generation of expanded, immunologically functional, CD4+ lymphocytes.


Major histocompatibility complex located complement C4 and steroid 21-hydroxylase gene rearrangements in couples with recurrent spontaneous abortions. Major Histocompatibility Complex (MHC) class III located complement C4 and steroid 21-hydroxylase (21OH) genes, which form various deletion and duplication units, were studied by TaqI Restriction Fragment Length Polymorphism (RFLP) in 58 Finnish couples who suffered recurrent spontaneous abortions (RSA). The gene rearrangements found in the RSA couples did not differ from those in the controls.


Restriction fragment length polymorphism analysis of zoo animals using HaeIII and four single-locus probes. Using HaeIII as the restriction endonuclease, restriction fragment length polymorphism analysis of dried blood samples from various animals was conducted. Single-locus probes D2S44, D10S28, D1S7, and D4S139, as well as monomorphic probe D7Z2, were used to examine for banding patterns. If bands were present, the samples were further examined for heterogeneity (whether single or multiple bands were observed) and polymorphism (whether variation in band location was shown between the animals studied within a species). Blood samples from animals, including primates, were obtained from Miami Metrozoo, Miami, Florida. Some of the animals were non-related individuals while others were related. Banding patterns were observed in colobus' for D2S44, D1S7, and D4S139; owl monkeys for D2S44; gorillas for D2S44 and D4S139; gibbons for D2S44 and D4S139; siamangs for D2S44, talapoins for DiS7; cranes for D1S7; and otters for D1S7. Based upon these, all of the animals for which a conclusion could be drawn appeared to be homozygous and monomorphic (exhibited only an invariant single band) for the loci examined except colobus' for D4S139, gorillas for D4S139, cranes for D1S7, and otters for D1S7.


Biochemical and haematological markers of alcohol intake in Ghanaians. The serum gamma-glutamyltransferase (GGT), aspartate aminotransferase (AST), urate and triglyceride and mean cell volume (MCV) were determined in 60 total abstainers, 56 social drinkers and 100 alcoholics. Both enzymes and urate showed progressive rise with increasing alcohol intake. The mean cell volume was only moderately elevated. Gamma-glutamyltransferase (GGT), aspartate aminotransferase (AST), and urate are sensitive enough to detect people who take in alcohol regularly and yet may be regarded as normal and not alcohol dependent.


Effect of operating variables on the separation of DNA molecules by capillary polyacrylamide gel electrophoresis. The influence of operating conditions on the separation of double stranded DNA molecules in capillary polyacrylamide gel electrophoresis is presented in continuation of previous work. In this study, an equation was derived that relates the effects of electrophoresis variables such as gel concentration, field strength, molecular weight and temperature to the migration velocity. As a model system, phage phi X174 DNA restriction fragments obtained by digestion with HaeIII have been examined. As an illustration of optimized conditions, DNA molecules up to the 10(3) base pair range were separated at high temperature (50 degrees C) in less than 6 min and with efficiencies as high as 5000 plates m-1 s-1.


Isolation and characterization of type 1 fimbriae from a chicken pathogenic Escherichia coli serotype O78. Type 1 fimbriae from chicken pathogenic Escherichia coli strain PDI-386 (serotype O78) was purified and characterized. Because of the acid-induced autoagglutination (T. Sekizaki, Y. Nakasato, and I. Nonomura, J. Vet. Med. Sci. 54, 493-499, 1992), the fimbriae could be easily purified by repeating acid sedimentation, washing, and dissolving in buffer (pH 8.0). In electron microscopy, the purified fimbriae showed a filament of 8 nm in diameter and 10 microns in average length. The molecular mass of the protein subunit of the purified fimbriae estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 19,000 daltons. The amino acid composition and its NH2-terminal sequence were similar to the previously described one of the Klebsiella pneumoniae type 1 fimbriae. Moreover, there was an immunological relatedness between them. These results indicated that a molecular diversity found between the fimbriae of E. coli and that of K. pneumoniae has already been existed among chicken pathogenic E. coli strains.


Increase of methionine aminopeptidase activity in hyperplastic Leydig cells of rat cryptorchid testis: a histochemical study. Histochemical study on the changes of the aminopeptidase activities in rat testes after surgically-induced cryptorchidism was conducted comparing them with the histochemical changes in regenerated hepatic cells of the partially hepatectomized rat liver. Methionine-aminopeptidase in Leydig cells gradually increased after cryptorchid was induced, whereas the enzyme activity in regenerated hepatic cells decreased. These histochemical observations were coincident with the data obtained by enzyme assay. The present study has indicated that in the rat cryptorchid testis the increase of methionine-aminopeptidase activity was caused by hyperplastic Leydig cells.


Antagonism of medetomidine sedation by atipamezole in pigs. The efficacy of atipamezole as a medetomidine antagonist was evaluated in pigs. The atipamezole doses (intramuscularly) were 80, 160, 320 and 480 micrograms/kg of body weight, which were one, two, four and six times higher than the preceding medetomidine dose (80 micrograms/kg, intramuscularly). Atipamezole effectively reversed medetomidine-induced sedation, and the optimal action was seen at doses of 160 and 320 micrograms/kg. Recovery from sedation was quick and smooth, and adverse effects such as hyperactivity or tachycardia were minimal with either dose.


Initial studies of embryonic transplants of human hippocampus and cerebral cortex derived from schizophrenic women. Human fetal brain tissue was obtained from first-trimester elective abortions of two women who also had schizophrenia. Portions of the embryonic hippocampus or cerebral cortex were transplanted into the anterior eye chamber of immunologically compromised athymic nude rats. In this environment, embryonic brain tissue derived from normal women generally continues organotypic growth and development for many months. Although initial survival after transplantation was normal, the tissue derived from schizophrenic women manifested less robust growth. However, cells in the transplants showed typical neuronal differentiation, with development of different neuronal types, such as pyramidal cells, granule cells, and gamma-aminobutyric acid (GABA)-containing interneurons. Rhythmic electrical activity was also observed, indicative of some local synaptic organization. The presence of messenger RNA (mRNA) for brain-derived neuronotrophic factor (BDNF) was observed using in situ hybridization. The reason for the decreased rate of growth of these transplants remains unknown and the significance of the finding cannot be assessed from only two fetuses. However, these preliminary findings suggest that fetal transplants may be a useful model system for the detection of developmental pathogenic processes in the expression and transmission of schizophrenia.


Assessment of myocardial perfusion, metabolism and pharmacology using positron emission tomography. Positron emission tomography is a non-invasive radionuclide imaging technique which enables in vivo assessment of regional cardiac function. When used with appropriate radiotracers and kinetic models, it can provide quantitative measurements of myocardial perfusion, metabolism and pharmacology. The development of quantitative measurements should help to decrease much of the ambiguity in the interpretation of data that often occurs. The short half-life of the radionuclides used allows the imaging of more than one function to be performed conveniently within the same scanning session with a low radiation dose to the subject. This integrated approach provides a means for investigating in vivo the physiology and pathophysiology of the human heart.


Angiotensin II-induced proliferation of cultured murine mesangial cells: inhibitory role of atrial natriuretic peptide. Angiotensin II (ANG II), as a single factor, induces proliferation in a cultured murine mesangial cell line (MMC). This study was undertaken to evaluate a possible influence of atrial natriuretic peptide (ANP) on this ANG II-induced proliferation. ANP (10(-7) M) for 2 min significantly increased intracellular cGMP levels in MMC. This increase in cGMP was totally abolished when cells were preincubated for 5 min with 10(-7) M ANG II. Stimulation of intracellular cGMP formation by sodium nitroprusside was also decreased in the presence of ANG II. The ANG II-mediated inhibition of ANP-stimulated intracellular cGMP levels was blocked by Dupont 753, suggesting signal transduction through ANG II receptors of the AT1 class. ANP (10(-7) M) for 24 h completely abolished the ANG II-induced proliferation in MMC. However, 10(-7) M ANP had no significant effect on mitogenesis induced by platelet-derived growth factor or epidermal growth factor. Furthermore, ANP reduced the ANG II-stimulated expression of the proliferating cell nuclear antigen, a cofactor of polymerase delta that is active in the S-phase of the cell cycle. The addition of 10(-3) M N-monobutyryl-guanosine 3':5'-cyclic monophosphate or 8-bromo-guanosine 3':5'-cyclic monophosphate also blocked the ANG II-induced proliferation. ANP (10(-7)) M for 24 h had no significant influence on the expression (number and dissociation constant) of ANG II receptors as determined by binding assays. These results suggest that, besides the previously shown vasoconstrictive and vasodilating effects, complex interactions between ANG II and ANP exist that can modulate mesangial cell growth.(ABSTRACT TRUNCATED AT 250 WORDS)


[Tropaphen in experimental drug therapy of cor pulmonale]. The influence of alpha-adrenoblocking drug tropaphen on cardio- and haemodynamics under experimental lung-heart failure was studied in dogs. The findings suggest effectiveness of tropaphen administration in such pathology shown by a decrease of lung hypertension and heart overload, significant increase of the contractile activity and relaxation properties of myocardium of both ventricles and in cardiac pump function optimization.


Simultaneous transcription of eleven cytokines in human alloreactive T lymphocyte clones after stimulation by phorbol ester and A23187. Human alloreactive T lymphocyte clones derived from cells invading a rejected kidney allograft, were analyzed for their ability to transcribe eleven cytokine genes under phorbol ester (PMA) plus calcium ionophore (CaI A 23187) stimulation. In addition to the positive signal previously obtained for IL-2 transcripts, strong specific patterns were seen with cytoplasmic dot hybridizations for IFN gamma and GM-CSF mRNAs in all the 17 clones screened. For the remaining transcripts (IL-3, IL-4, IL-5, IL-6, TNF alpha, LT, M-CSF and HILDA/LIF), these techniques proved to be inadequate. Northern-blots were therefore performed on three clones exhibiting different phenotypes (CD4+ CD8- non cytotoxic, CD4+ CD8- cytotoxic and CD4- CD8+ cytotoxic). Positive specific signal with the eleven probes could be obtained. Nevertheless, the IL-6 message was found only in the helper clone and the TNF alpha transcript appeared at a later time point compared to the other cytokine messages (its maximum expression was observed around 24 hours post-stimulation). In conclusion, we demonstrate that under PMA+CaI activation, one clone is able to simultaneously transcribe at least eleven lymphokine genes. Except, perhaps for IL-6, the pattern of lymphokine transcription did not permit us to distinguish between different lymphocyte subsets.


Didanosine therapy in patients intolerant of or failing zidovudine therapy. To examine the effect of didanosine (2',3'-dideoxyinosine, ddI) on surrogate markers of HIV infection (CD4+ lymphocyte count, p24 antigen) and to evaluate the incidence of adverse effects from ddI. This study was performed as a retrospective chart review of patients who were enrolled in Bristol-Myers Squibb's expanded-access program for ddI. Patient records were obtained from primary care physicians' offices. Twenty-five HIV-infected patients diagnosed with AIDS or AIDS-related complex (ARC) who were intolerant of zidovudine (ZDV) therapy or deteriorating clinically despite ZDV therapy and were eligible for inclusion in the ddI expanded-access program. ddI was administered orally in a citrate-phosphate buffer every 12 hours. Patients were followed by their private physician on a monthly basis. Laboratory analysis at each month included CD4+ lymphocyte count, hemoglobin, hematocrit, serum amylase, uric acid, serum triglycerides, and p24 antigen. Mean CD4+ cell count, serum amylase, hemoglobin, and uric acid at each month during ddI therapy were compared with baseline concentrations for nine months. Patients had received prior ZDV therapy for an average of 15.5 months before starting ddI. Mean CD4+ cell counts increased from 53.9/mm3 at baseline to 72.4/mm3 after 4 months of therapy (p = 0.04) but returned to concentrations comparable with those at baseline after 5 months. One case of documented pancreatitis, two cases of suspected pancreatitis, and nine cases of peripheral neuropathy occurred during ddI therapy. The estimated mean cumulative dose for the development of neuropathy was 1.16 g/kg, which is lower than previously reported. Patients who have received prolonged therapy with ZDV or who have low initial CD4+ counts may not have sustained increases in CD4+ counts from ddI therapy. Also, development of peripheral neuropathy may occur at lower cumulative doses in these patient populations.


Identification of the B cell derived T cell colony promoting activity to soluble CD23. We previously reported that supernatants of phytohemagglutinin (PHA)-stimulated normal human B cells (NBCsup) contain a T cell colony promoting activity. NBCsup were able to (a) increase the number of secondary colonies generated under PHA and interleukin-2 (IL-2) stimulation by peripheral blood-derived primary T colony cells, (b) enhance the ability of CD4+ but not CD8+ peripheral blood T cells to form agar colonies in the presence of PHA and IL-2 and (c) support in vitro differentiation of CD2-3-4-8- prothymocytes into CD2+3+4+ T cells. This activity was therefore refered to as Prothymocyte Differentiating Activity (PTDA). Subsequent studies pointed to striking biochemical and cell source homologies between this B cell derived factor and the 25-kDa soluble CD23 (sCD23). sCD23 has been recently found to display prothymocyte differentiating activity.


Adjunctive medications in patients receiving thrombolytic therapy: a multicenter prospective assessment. The Virginia Multicenter Thrombolytic Study Group. To describe the use of adjunctive therapies in patients with acute myocardial infarction receiving thrombolytic agents. Data were collected prospectively by the study-site investigator or the emergency department physician caring for the patient. Study participation did not influence thrombolytic regimen selection or the adjunctive therapies ordered. Thirteen Virginia hospitals representing a cross-section of hospitals in the state. Eleven are urban medical centers; four have graduate medical education programs. Patients were included in the study if the decision to administer thrombolytic therapy was made in the emergency department. Concomitant medications administered during the first six hours after initiation of thrombolytic therapy. Two hundred ten patients (aged 57 +/- 14.1 y) were evaluated. Ninety-five percent of these patients were treated with tissue plasminogen activator, 3 percent received anisoylated plasminogen streptokinase activator complex, and 2 percent received streptokinase. Ninety-one percent of the patients also received heparin, the most commonly used adjunctive medication; 77 percent concomitantly received lidocaine; 62 percent received aspirin; and only 19 percent received a beta-blocker. Our data provide a reference point for future studies to determine factors that influence the selection of adjunctive agents for treating patients with acute myocardial infarction receiving thrombolytics.


Nonneuroleptic treatment of disruptive behavior in organic mental syndromes. To summarize the literature describing nonneuroleptic treatments of unacceptably disruptive behavior in chronically institutionalized psychiatric patients with mental retardation, autism, organic brain syndrome, and dementia. Relevant articles were identified from a MEDLINE search of the above diagnoses linked with "aggression" and "psychomotor agitation." Additional references were found in the bibliographies of these articles. The studies reviewed were limited to prospective evaluations of nonneuroleptic drug therapy of these behavior disturbances. Case reports, case series, and retrospective studies were excluded. Studies of patients with schizophrenia, affective disorders, and personality disorders were also excluded. Studies of lithium, beta-blockers, carbamazepine, benzodiazepines, and buspirone were adequate for review. As a rule, these studies are hampered by poor design. The lithium studies suggest that mentally retarded patients with behavior disturbances may respond to lithium treatment. The beta-blocker studies suggest improvement in patients with mental retardation, autism, organic brain syndrome, and dementia. Neuroleptic discontinuation or a decrease in dose was possible in some patients. The carbamazepine studies were inconclusive. Carbamazepine should be reserved for patients with concomitant seizure disorders. Benzodiazepines were helpful in treating elderly demented patients. Thus far, buspirone has been evaluated in only a few, poorly designed studies and is not yet recommended for treatment of behavior disturbances. Legislation has restricted the use of neuroleptics in many patients receiving long-term healthcare. Despite the questionable validity of the studies reviewed, lithium, beta-blockers, carbamazepine, and benzodiazepines may be considered as alternatives to neuroleptics in selected cases.


Evaluation of the reliability and validity of a measure of anxiolytic drug-use intensity for pharmacoepidemiologic studies. To assess the reliability and validity of a proposed new standard of drug-use intensity, the minimum marketed dose (MMD), using anxiolytic drugs as models. Retrospective, cohort design. Staff model, nonprofit health maintenance organization. Eighty-five patients who obtained one or more prescriptions for an anxiolytic, antidepressant, or sedative-hypnotic drug during three consecutive one-year time periods. Fifty-nine patients had a prescription filled during the year before the study. For anxiolytic drugs, the magnitude of the average correlation for the MMD measure of drug-use intensity was greater than the total number of prescriptions and the total number of dosage units. Discriminant validity was demonstrated because the MMD was not correlated with measures in unrelated therapeutic categories. Summed MMD units were shown to significantly predict physical impairment (criterion validity). The MMD measure of drug-use intensity was reliable and valid for anxiolytic drugs. These findings suggest that information gathered from automated prescription records may be a useful indicator of drug-use intensity in pharmacoepidemiologic studies.


Differentiation of isolates of Discula umbrinella (Teleomorph: Apiognomonia errabunda) from beech, chestnut, and oak using randomly amplified polymorphic DNA markers. Genetic variation in 30 isolates of Discula umbrinella derived from beech, chestnut, and oak was assessed using randomly amplified polymorphic DNA (RAPD) and restriction fragment length polymorphic markers. Polymerase chain reaction amplifications with 17 primers produced 134 different DNA fragments. Three RAPD fragments were subsequently used for Southern hybridization. By these techniques up to four different individuals could be detected in the same leaf. The presence of several individuals within a single leaf indicates a finely tuned balance between the endophyte and its host. Cluster analysis of all arbitrary primed amplified DNA fragments showed that the isolates could be placed into four groups corresponding to their host origin. The high percentage of private RAPD variants within groups is consistent with low gene flow.


Increased frequency of the heterozygous switch region of IgA2 in Japanese patients with IgA nephropathy. The relation between IgA hyperproduction and restriction fragment length polymorphism (RFLP) of the immunoglobulin heavy chain switch (S) region was examined in Japanese patients with IgA nephropathy (IgAN) using Southern blot hybridization. Polymorphism in the S regions of IgM, IgA1 and IgA2 (SA2) was detected. A significant increase in the frequency of heterozygous phenotype of SA2 was shown in the patients. These patients showed a significant increase in the amount of serum IgA, IgA bearing cells and levels of proteinuria. Although two reports on RFLP of the S region have been published in Europe, the results differed. A comparison of the three reports showed different frequencies in the phenotype of the S region between Caucasian and Japanese patients. These results suggested that there is heterogeneity at the S region in IgAN patients in various countries and that this polymorphism might be associated with IgA hyperproduction and the development of proteinuria in Japanese patients.


Beta(+)-thalassemia with hemochromatosis. A 64-year-old man was admitted due to ascites. Laboratory data showed hemoglobin 6.7 g/dl, mean corpuscular volume 82 fl, and ferritin 2,360 ng/ml. Liver biopsy showed hemochromatosis. The diagnosis of beta-thalassemia was suggested by a decreased ratio of beta/alpha-globin synthesis in vitro (0.26). Cloning of the beta-globin gene showed A-to-G mutation in the first base of the ATA box. He was confirmed to be homozygous for this specific allele by beta-gene complex analysis and analysis of Southern blot hybridization of the alpha- and beta-globin genes. His two sons were confirmed to be heterozygous for this allele.


Genetic analysis of systemic lupus erythematosus: association with a T cell receptor restriction fragment length polymorphism in Japanese patients. Many studies have suggested the involvement of multiple genetic loci in the development of systemic lupus erythematosus (SLE). We have analyzed the correlation between various genetic markers and the susceptibility to SLE. In this study, the association of SLE and restriction fragment length polymorphism (RFLP) of T cell receptor gene was evaluated. The cDNA for constant regions of alpha, beta and gamma chain were used as probes and RFLPs were analyzed after digestion with Eco RI, Bam HI, Pst I, Pvu II, Hind III and Bgl II. Among them, polymorphisms were detected using Bgl II- and Hind III-digested DNA and C beta as a probe. Association with SLE in Japanese patients was found only after digestion with Hind III. The absence of the 13 kb polymorphic band appeared to be correlated with the development of SLE (relative risk = 4.78).


Fine-structure mapping of the complex locus Odc-rs5 relative to Igk and distal loci. Odc-rs5 was previously identified as a complex locus closely linked to the Igk complex on mouse Chromosome (Chr) 6 and comprising at least five copies of a sequence related to the mRNA encoding ornithine decarboxylase (ODC) in the genomes of mice of some inbred strains and at least seven copies in others (Richards-Smith and Elliott, Mammalian Genome 2: 215, 1992). In the present study, Odc-rs5 was shown to be composed of at least seven copies of the ODC sequence in both the Odc-rs5a and Odc-rs5b haplotypes. Based upon the distribution of DNA restriction fragments (RFs) that had previously been associated with Odc-rs5a or Odc-rs5b among 42 mice of inbred laboratory strains having various haplotypes at Igk and in mice of two congenic strains [B6.PL-Ly-2a, Ly-3a(75NS)/Cy and B6.PL-Ly-2a,Ly-3a(85NS)/Cy] and a backcross-derived stock (NAK) known to be recombinant within Igk, a fine structure map of Odc-rs5 was deduced relative to Igk and more distal loci. Odc-rs5-derived RFs were located to three distinct regions within and/or distal to Igk and to a fourth site between (Ly-3, Ly-2) and Raf-1. Additionally, DNAs from 19 mice of inbred strains and random-bred stocks derived from wild progenitors trapped at various locations were analyzed and found to exhibit an unexpected variety of combinations of RFs associated with the two Odc-rs5 haplotypes most frequently observed among inbred laboratory strains of mice.


Theophylline produces over-additive relaxation of canine tracheal smooth muscle when combined with beta-agonists: the dose-response relationship. The interaction between theophylline (T) and the beta-agonists albuterol (A) and isoproterenol (I) was examined using canine cervical tracheal smooth muscle devoid of epithelium contracted with 0.1 or 0.3 microM methacholine. Greater functional antagonism with beta-agonists vs. T was confirmed and an ability of T to potentiate beta-agonist relaxation was demonstrated. The EC50 for T increased from 0.13 +/- 0.02 to 0.37 +/- 0.07 mM (mean +/- SEM) in preparations contracted with 0.1 or 0.3 microM methacholine, respectively, while that for I increased from 0.036 +/- 0.008 to 0.17 +/- 0.03 microM, a significantly larger change (P < 0.025). In tissues contracted with 0.3 microM methacholine and pretreated with 10 micrograms/ml of T IC50 values from composite concentration-response curves for I and A were displaced to the left and Emax was increased (56.6 to 71.5% for I, 44 to 61% for A, P < 0.0002). Addition of 10 micrograms/ml T resulted in relaxations which exceeded that calculated by the fractional product method for additive, independent action (P < 0.0001 for I, P < 0.0002 for A at 0.3 microM methacholine), suggesting that at least part of T's action was over-additive. Five, 10 and 20 micrograms/ml T enhanced the effectiveness of single concentrations of I by factors of 1.47 +/- 0.14 (P < 0.05), 2.72 +/- 0.26 (P < 0.01) and 5.34 +/- 0.55 (P < 0.01), respectively, in preparations contracted with 0.1 microM methacholine: I enhanced the effectiveness to a lesser degree. Using two approaches, positive interaction or over-additivity between T and beta-agonists has been demonstrated.


The effects of salmeterol and salbutamol on ciliary beat frequency of cultured human bronchial epithelial cells, in vitro. Studies investigating mechanisms of mucociliary clearance have suggested that beta 2-adrenergic agents may significantly influence ciliary activity of epithelial cells and therefore play a vital role in the maintenance of functional integrity of the airways. We have cultured human bronchial epithelial cells, from surgical explants and investigated the effects of salbutamol and salmeterol, in a time- and dose-dependent manner, on the ciliary beat frequency (CBF) of these cells. Prior to and at several times after exposure to either salbutamol (10(-8) to 10(-3) M) or salmeterol (10(-8) to 10(-4) M), the epithelial cells were monitored for CBF and on the basis of data obtained from these studies, the effect of 10(-6) M propranolol was investigated in the presence of optimal concentrations of salbutamol and salmeterol. Salbutamol was optimally active at a concentration of 10(-4) M and caused a transient but significant increase in the CBF from baseline level of 8.6 +/- 0.4 to 9.6 +/- 0.5 Hz (P < 0.05), after 2 h incubation. In contrast, salmeterol was maximally active at a concentration of 10(-6) M and caused a significantly rapid and prolonged increase in CBF from a baseline value of 9.2 +/- 0.4 to 10.9 +/- 0.6 Hz (P < 0.02) and 10.6 +/- 0.8 Hz (P < 0.05) after 15 min and 24 h incubation, respectively. Propranolol (10(-6) M) abrogated the salbutamol- but not the salmeterol-induced increases in CBF.(ABSTRACT TRUNCATED AT 250 WORDS)


Pattern generation.

Pattern generation. The study of rhythmic motor pattern generation continues to be dominated by preparations in which cellular and circuit mechanisms can be bridged. Using these preparations, basic questions such as how circuits can be modified in order to generate a large number of patterns have begun to be answered. The action of neuromodulators on second messengers and channel proteins can provide a link between molecular studies and behavior. There has been a large increase in the use of computer simulations, and their usefulness as a way of studying pattern generation is growing.


Mechanisms of respiratory rhythm generation. In mammals, a three-phasic respiratory rhythm is generated by a network of various types of neurons in the lower brainstem. The cellular mechanisms of rhythmogenesis involve cooperative interactions between synaptic processes and specific membrane properties. The network seems to be driven by extrinsic sources in mature animals, whereas in the immature network pacemaker neurons might be involved.


Control of functional systems in the brainstem and spinal cord. Progress has been made in the identification of cells, circuits, and networks involved in certain important subcortical functional systems, including swallowing, chewing, posture and locomotion, and in the shared mechanisms for selecting the network for specific motor tasks, including a role for excitatory amino acids for network activation, the shaping of the network by inhibitory control, and the selection of inputs and modulation of outputs by monoamines and other agents.


Toxic effect of bile acid ingestion in rats. Ingestion of raw bile from some grass carp fish has been reported to be associated with impairment of renal function in some Chinese people. 5 alpha-cyprinol, the main bile alcohol in these fish, has been suggested as the toxic compound causing electrolyte imbalance. In this study, we attempted to determine whether oral administration of pure bile acids also affects the electrolyte balance in rats. Twenty-four female Long-Evans rats weighing 200-250 g were used. Conscious cannulated animals were prepared and studied with gastric intubation of 15%, 4.8 mL/kg of sodium cholate (SC), sodium deoxycholate (SD), and water; there were three groups of eight rats in each. A drastic drop in blood pressure, elevation of the ST segment of the EKG, increases in plasma potassium, hydrogen, blood urea nitrogen, and oxygen partial pressure were found. All rats treated with bile acids died within 24 hours. Six SD rats and three SC rats died within eight hours after oral administration of bile acid. It is concluded that the effects of feeding rats bile acids are very similar to the toxic action seen after ingestion of grass carp bile.


Migration of peptide-immunoreactive local circuit neurons to rat cingulate cortex. The times of origin of neurons immunoreactive for somatostatin (SRIF), cholecystokinin (CCK), and vasoactive intestinal polypeptide (VIP) were determined using a combined immunohistochemical-autoradiographic technique. 3H-thymidine (3H-dT) was injected into pregnant rats on gestational day 13 (G13), G15, G17, G19, or G21. Animals were killed on postnatal day 30, that is, after the completion of neuronal migration. Sections of the brain including posterior cingulate cortex (area 29), visual cortex (area 17), and somatosensory cortex (area 3) were processed serially for peptide immunoreactivity and autoradiographically for labeling with 3H-dT. SRIF- and CCK-immunoreactive neurons were cogenerated and comigrated according to an inside-to-outside sequence. Accordingly, neurons in layer VI were born on G13, neurons in intermediate layers were born on G15-G17, and neurons in layer II/III were born on G19-G21. In contrast, VIP-positive neurons did not follow such a sequence. Neurons in all layers of cortex were generated at relatively constant rates between G13 and G21. VIP-immunoreactive neurons were the only known subpopulation of neurons that did not migrate into cortex by an inside-to-outside sequence. Thus, the migration of local circuit neurons to cingulate cortex follows patterns that are similar to those discerned in more differentiated cortical areas.


Generation of neutrophil-chemoattractant by degradation of tumor cell membrane protein. Polymorphonuclear leukocytes (PMN) infiltrating into tumors are assumed to be the result of a migration in response to a tumor-derived chemotactic factor. It is hypothesized that tumor-derived chemotactic factors, which are supposed to be in aggregates, can be exposed by proteolytic degradation of the tumor cell membrane protein (TCM). The TCM suspension made from either Sarcoma 180 or Ehrlich ascites tumor after incubation with Varidase at 37 degrees C for six to eight weeks is referred to as a degraded TCM protein (DTCM). The purpose of this study was to detect whether or not the DTCM was chemotactic for PMNs. Chemotactic activity was assayed by a sponge-matrix model and expressed in the number of PMNs infiltrating into the sponge per mouse after intrasponge injection of the DTCM, dose range, 0.4-0.005 mL. Maximal response to DTCM was restricted to the limited dose between 0.01 and 0.02 mL. Quantities of DTCM > 0.02 mL caused a response in roughly reciprocal proportion to the dose of DTCM. Changes in the circulating leukocyte count in response to an intravenous (i.v.) injection of DTCM was biphasic. The initial relative leukopenia was followed by leukocytosis within 24 hours in rats surviving the effect of DTCM doses < 0.5 mL. However, in rats with i.v. injections of higher doses, profound leukopenia was present at death. The mice, after an intraperitoneal (i.p.) injection of DTCM, dose range, 0.5-1.0 mL, died between 16 and 24 hours after the injection. The bone marrow of these mice showed complete depletion of PMNs.(ABSTRACT TRUNCATED AT 250 WORDS)


Adult bacteremic pneumonia: bacteriology and prognostic factors. Bacteremic pneumonia is a highly specified subgroup of pneumonia that is potentially life-threatening. In order to find out the prognostic factors in this subgroup of pneumonia, we conducted a 40-month retrospective analysis of 70 cases in our hospital. The male to female ratio was 54:16. Forty-one cases were community-acquired bacteremic pneumonia (CABP), and 29 cases were nosocomial bacteremic pneumonia (NBP). Both CABP and NBP were predominated by gram-negative bacteria. Klebsiella pneumoniae was the most common microorganism isolated in both CABP and NBP. The overall mortality was 62.9% (44/70). There was no significant difference in the mortality between CABP (61.0%) and NBP (65.5%). After univariate analysis of all possible prognostic factors, 10 variables were found to have significantly poor prognostic values. They were: 1) the presence of septic shock; 2) the use of ventilatory support; 3) the presence of radiologic spread; 4) treatment in an intensive care unit; 5) male gender; 6) the development of adult respiratory distress syndrome; 7) Klebsiella bacteremic pneumonia in patients with an alcohol habit; 8) patients with ultimately fatal underlying diseases; 9) an initial AaDO2 > 200 mmHg; and 10) an initial arterial pH < 7.25.


Serum sialyl stage-specific embryonic antigen levels in adenocarcinoma of the lung. The serum levels of sialyl stage-specific embryonic antigen (SSEA-1) in 67 patients with adenocarcinoma of the lung were studied to assess their values for diagnosis. A solid-phase immunoradiometric sandwich assay with an FH6 monoclonal antibody was used. Another 49 healthy adults, 52 patients with benign pulmonary diseases, and 142 lung cancer cases with cell types other than adenocarcinoma were evaluated for comparison. The mean (+/- S.D.) levels (U/mL) for adenocarcinoma of the lung, lung cancer other than adenocarcinoma, benign pulmonary diseases and normal subjects were 182.9 +/- 309.7, 53.5 +/- 22.6, 38.9 +/- 17.1 and 30.5 +/- 6.5, respectively. The mean serum sialyl SSEA-1 level was significantly higher in adenocarcinoma of the lung, compared with lung cancer other than adenocarcinoma (p < 0.001), benign pulmonary diseases (p < 0.001), or normal subjects (p < 0.001). For late stage (Stages III and IV, n = 58) adenocarcinoma of the lung, the mean (+/- S.D.) serum sialyl SSEA-1 level (204.3 +/- 327.6 U/mL) was significantly higher than that of earlier stage (Stages I and II, n = 9) adenocarcinoma of the lung (39.9 +/- 19.3), p < 0.001. There was no statistical difference among the mean serum levels of various histologic types of lung cancer other than adenocarcinoma (p > 0.05). In the lower range of values, considerable overlap existed between adenocarcinoma of the lung and lung cancer other than adenocarcinoma. However, very high sialyl SSEA-1 levels (> 140 U/mL) were only encountered in late stage adenocarcinoma of the lung (22/58).(ABSTRACT TRUNCATED AT 250 WORDS)


Prognosis of thymic carcinoma: analysis of 16 cases. Primary thymic carcinoma is a rare malignant neoplasm which arises from the thymic epithelium. Between May 1982 and September 1990, 16 patients with thymic carcinomas, diagnosed at Chang Gung Memorial Hospital, were reviewed. Their ages ranged from 19 to 75 years, with a median of 40 years. Males predominated (male to female ratio was 11:5). The most common presenting symptoms were chest pain, cough, body weight loss and dyspnea. No paraneoplastic syndromes were seen. Chest roentgenograms of 15 patients revealed a mediastinal mass, but a definitive diagnosis could not be made until surgery. Adjacent mediastinal tissues were invaded, or adhered to, by all the tumors. Six distinct histologic types were found, squamous cell carcinoma being the most common (seven cases). The primary treatment of surgical resection was attempted in 14 patients, but only in five cases could the tumors be completely resected; two had a biopsy only. Radiotherapy, with or without chemotherapy, given postoperatively, achieved additional local control in seven of the nine partially resected patients (77%). Distant metastasis occurred in nine of 16 patients (56%). Lymph nodes, bone and lung were the most common metastatic sites. Chemotherapy with cisplatin and/or adriamycin-based regimens was given to patients who had distant metastasis, but the responses were unsatisfactory. The overall survival at one, three and five years was 88%, 51% and 31%, respectively. The median survival was 30 months. The median survival of patients with pure squamous cell carcinoma (> 49 months) was superior to that of patients with other histologic types (18 months; p < 0.01).


Risk factors for hyperinsulinemia in chlorpropamide-treated diabetic patients: a three-year follow-up. To elucidate the presence of chronic hyperinsulinemia and its relation to clinical and biochemical parameters, 112 (53 females and 59 males) Chinese non-insulin-dependent diabetes mellitus (NIDDM) patients under chlorpropamide therapy were closely monitored for three years. Clinical and biochemical risk factors for chronic hyperinsulinemia were studied by regular monitoring of body weight, fasting insulin levels and various biochemical data. Chronic hyperinsulinemia was defined as a mean fasting level over 20 microU/mL (highest level observed in 35 non-diabetics). Among 112 diabetics, 52 cases (46.4%) showed chronic hyperinsulinemia. From simple linear regression analysis, female gender, high BMI and elevated triglyceride and uric acid levels were correlated with insulin levels (p < 0.05). The presence of diabetic complications (retinopathy, neuropathy and nephropathy) and the degree of glycemic control were not significantly different between the normoinsulinemic and hyperinsulinemic groups. In conclusion, 1) NIDDM patients treated with chlorpropamide showed higher fasting insulin levels with 46.4% of them meeting the criteria for chronic hyperinsulinemia; 2) female gender, uric acid, BMI and triglyceride were the risk factors correlated with chronic hyperinsulinemia; and 3) the presence of diabetic complications and diabetic control correlated poorly with chronic hyperinsulinemia.


Plasma prorenin and renin levels in non-insulin-dependent diabetes mellitus. To investigate plasma renin and prorenin levels in non-insulin-dependent diabetes mellitus (NIDDM) and their relation with autonomic nervous function and renal impairment, we measured plasma renin and prorenin levels in 39 NIDDM patients. The patients included 21 males and 18 females, aged 56.3 +/- 6.2. Thirty-four normal age-matched subjects served as controls. Autonomic nervous function was evaluated in 23 patients by the performance of cardiovascular reflex tests. The plasma renin concentration was measured by angiotensin I generation after the addition of an exogenous substrate. Plasma prorenin was activated by trypsin. The results showed that the plasma renin concentration was similar between NIDDM patients and normal subjects, while plasma prorenin was higher in NIDDM patients. No correlation existed between the plasma renin or prorenin levels and autonomic nervous function. The patients with abnormally high levels of prorenin also had a similarly high plasma renin level but not a high creatinine clearance (Ccr) or daily proteinuria. The plasma renin level was correlated inversely with daily proteinuria but not with Ccr. These results suggest that the high plasma prorenin levels in some diabetic patients cannot be explained by renal impairment, poor prorenin conversion or autonomic dysfunction. The hyporeninemia in some patients may be related to microvascular involvement of the kidney.


Energy requirements in response to high protein feeding in young male adults. Two experiments were conducted to investigate the effects of moderate (MPI) and high (HPI) levels of protein intake on energy utilization in 12 healthy young male adults who were confined to a metabolic unit for 56 days. The amount of energy supplied was adjusted so that the subjects could maintain a relatively constant body weight, which was used as the criterion to see whether the body was under an energy balance. All foods were supplied in conventional style. In the first experiment, six young men were given a diet of 1.18 g (MPI) and 1.74 g (HPI) protein/kg of body weight/day in consecutive periods. In the second experiment, another six young men were studied at a protein intake of 1.08 g (MPI) and 2.00 g (HPI)/kg/day. The results indicate that both energy intake and energy expenditure are independent of protein intake. The average gross energy intake of the subjects was between 2,300 to 3,240 kcal/day, corresponding to a light-to-moderate grade of energy expenditure. The efficiency of energy utilization did not differ significantly between the moderate and high levels of protein intake in adults.


Complications of laparoscopic cholecystectomy: an analysis of 200 cases. Complications of the initial 200 cases of laparoscopic cholecystectomy (LC) at the Cathay General Hospital within a period of 11 months were reviewed from video documents of the operations and clinical records. The major complication rate was 3.5%, including one common bile duct (CBD) injury (0.5%), three retained CBD stones (1.5%), one subphrenic fluid accumulation (0.5%), one liver abscess (0.5%) and one cystic duct stump bile leakage (0.5%). All major complications were cholecystectomy-related, and only one of the seven occurred in cases of acute cholecystitis. Age and sex were not related to its occurrence. The rate of minor complications ranged from 0.5% to 10%; they were: shoulder and back pain (10%), gall bladder perforation (10%), retained stones in the abdominal cavity (5%), transient nausea and diarrhea (5%), extension of umbilical port to a mini-laparotomy (3.5%), prolonged operation time > three hours (2%), subcutaneous emphysema (1.5%), wound infection (1.5%) and prolonged ileus (0.5%). The minor complications occurred largely in patients with acute cholecystitis. The complications occurred mostly during the early period of our study, indicating a learning period phenomenon. These could have been avoided if we had had a thorough knowledge of the potential complications and had strictly followed the principles of laparoscopic surgery. We conclude that LC is safe and the complication rate is not higher than that for open cholecystectomy. Most of the complications are preventable if LC is performed by qualified biliary surgeons following strict precautions.


Comparison of omeprazole and nizatidine in the treatment of duodenal ulcers. This study compares the efficacy and side effects of omeprazole and regular- and double-dose nizatidine in the treatment of duodenal ulcers. Duodenal ulcer healing rates in these three groups (omeprazole, 20 mg qd; nizatidine, 300 mg hs and 600 mg hs) were 81.8%, 19% and 30%, respectively, after two weeks of therapy; and 90.5%, 70% and 84.2%, respectively, after four weeks of treatment. Omeprazole had a significantly better healing rate than nizatidine, 300 mg or 600 mg, after two weeks of treatment (p < 0.01), but not after four weeks of treatment. Omeprazole relieved the ulcer pain sooner than nizatidine (p < 0.05). Smoking decreased the duodenal ulcer healing rate in the omeprazole group, but not in the nizatidine groups. Clinical features, such as sex, age, alcohol consumption, ulcer size, past history of upper gastrointestinal bleeding and duration of peptic ulcer history, did not collate with the healing rate. Patients with double-dose nizatidine did not show any benefits over those with a regular dose in this study. Adverse effects were minor, and there were no significant changes in biochemistry after therapy in these three groups of patients.


In vitro perfusion study of the human placenta at term: preliminary results. Dual perfusion of the human placental lobule in vitro is a useful method for studying the transfer of molecules across the placenta, including the transfer of endogenous substances and xenobiotics. To establish the first model of in vitro placental dynamic study in our country, we used a dual recirculating perfusion system modified from that described by Miller et al. A placental lobule without tears or gross lesions was chosen. Both the fetal and maternal sides of the placenta were perfused with Medium 199 in addition to heparin, glucose, dextran and antibiotics. Perfusate samples were obtained periodically and analyzed for blood gas, glucose, lactate, human placental lactogen (hPL) and human chorionic gonadotropin (hCG). The stability of this human placental lobule preparation during 10 hours of perfusion was reflected by the stability of the arterial pressure and by the constant volume in the fetal compartment. A constant rate of oxygen was delivered by the maternal circulation system, and a steady rate of oxygen was gained by the fetal circulation system. Neither oxygen nor glucose consumption by the tissue was significantly reduced during the period of perfusion. The releasing rates of hCG and hPL did not change significantly during perfusion. The development of this dual perfusion system for the human placenta can provide for the study of placental hemodynamics and transplacental transport in perinatal medicine in our country.


Binding and growth-stimulation of cervical cancer cell lines by prolactin. Two cervical cancer cell lines CC7-T and Si-Ha were employed to observe the relationship between cervical cancer and prolactin. By immunocytochemical and indirect immunofluorescent assays using two prolactin monoclonal antibodies PRL-149 and PRL-151, both cell lines with added prolactin (10 ng/mL) were noted to be positive for PRL-151, but negative for PRL-149. The control cell lines from ovarian cancer and the myeloma lines were both stained negative. By using MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, it was noted that CC7-T and Si-Ha grew better in the presence of various added concentrations of prolactin, ranging from 0.1 to 1,000 ng/mL, suggesting that prolactin may enhance the growth of cervical cancer. The degree of stimulation appears to depend on cell differentiation. However, prolactin levels in the cultured supernatant were undetectable by the enzyme immunoassay (EIA) method. We postulate that prolactin can bind and stimulate the growth of some cervical cancer cell lines, probably through the prolactin receptor rather than by autocrine regulation.


Streptococcus bovis endocarditis associated with colonic adenocarcinoma: report of a case. Streptococcus bovis (S. bovis) endocarditis has been increasing over recent decades, especially among the senile population. A 74-year-old man presented with intermittent fever for two months. He had a past history of aortic dissection and underwent a Bentall operation one year before admission. A Janeway lesion was noted on his right hand and six blood cultures grew S. bovis. He was treated with penicillin-G, 3 microU intravenously, every six hours, and became afebrile three days later. A colonofiberoscopy was carried out despite the absence of any gastrointestinal symptoms, and a 2 x 2 cm mass was found at the cecum, with pathologic proof of adenocarcinoma. The patient died from a massive intracranial hemorrhage on the 23rd hospital day. Review of the literature revealed an intimate association between S. bovis bacteremia (or endocarditis) and underlying colonic neoplasia. Failure to be aware of the possible consequences of this combination may lead to detrimental patient prognosis. We strongly advise that every patient presenting with bacteremia or endocarditis due to this organism, even if they are free from gastrointestinal symptoms, should undergo a thorough lower gastrointestinal investigation to rule out colonic neoplasia.


Multiple cerebral aneurysms in a child with cardiac myxoma. Neurologic involvement is not uncommon in cardiac myxoma, yet its association with multiple cerebral aneurysms is rare, especially in children. We describe a patient with left atrial and ventricular myxoma, who otherwise lacked cardiac symptomatology and did not exhibit auscultatory, roentgenologic, or electro-cardiographic evidence of heart disease. The clinical presentation was a sudden attack of a cerebrovascular occlusive disorder. Multiple fusiform aneurysms at the frontal and parietal branches of the left anterior and middle cerebral arteries were demonstrated on cerebral angiograms. The myxomas were surgically removed and the patient improved.


Cardiac tamponade resulting from recurrent small-cell carcinoma of the uterine cervix temporarily responding to CE/CAV chemotherapy: report of a case. A case of recurrent small-cell carcinoma of the uterine cervix, initially presenting with cardiac tamponade, is reported. After pericardiotomy, the patient was treated with an alternating combination of chemotherapy, which included cisplatin plus etoposide (CE) and cyclophosphamide, adriamycin plus vincristine (CAV). A partial response, with relief of a cough and diminishing metastatic pulmonary lesions, was noted from serial chest roentgenographs after the initial three cycles of chemotherapy. The patient did not receive any further treatment and the recurrent cough and dyspnea were noted two months later. In spite of the same chemotherapeutic regimen and chest radiotherapy, the patient died nine months after the initial diagnosis of metastasis.


Enhancement of DNA synthesis in rat thymocytes by stimulating their muscarinic acetylcholine receptors. The binding of 3H-acetylcholine (ACh) to acetylcholine receptors (AChRs) on rat thymocytes was examined and found to be inhibited by the treatment with several antagonists against nicotinic and muscarinic AChRs. This result suggested that thymocytes have AChRs with different affinity, and bear both nicotinic and muscarinic AChRs on their surfaces. To make clear the functional significance of the AChRs, DNA synthesis of the thymocytes stimulated with ACh was examined. 3H-thymidine uptake of thymocytes was significantly increased when the cells were stimulated with ACh or another cholinergic agonist. The increment of DNA synthesis caused by ACh in thymocytes was not reduced by treatment with nicotinic antagonists, but was decreased by treatment with any of the muscarinic antagonists. Concentration of the intracellular second messengers, inositol 1,4,5-triphosphate (IP3) and guanosine 3',5'-cyclic monophosphate (cGMP) was also made higher by ACh stimulation. It is discussed that the enhancement of intracellular IP3 and cGMP concentrations after stimulation of muscarinic AChRs appears to be related with the increment of thymocyte DNA synthesis.


Modeling of heteroduplex formation during PCR from mixtures of DNA templates. We have investigated the capability of PCR to amplify a specific locus from each template in a mixture of allelic DNAs. Modeling experiments employed a 300-bp HOX2B segment as target and utilized, as distinct template "alleles," genomic DNAs from 1 human and from 2 chimpanzees known to differ in sequence at the target. Two modes were used: (1) mixtures of PCR products that had been previously amplified individually from a single template and (2) PCR amplification en masse from composite human/chimpanzee genomic DNA templates. Products generated by either mode were separated by denaturing gradient gel electrophoresis (DGGE). Detection of a "trace" allele mixed with a "dominant" one was possible by simple ethidium bromide staining of the gel up to a sensitivity of 1 part in 20. A balanced mixture was represented by 5:5 and 4:3:3 mixtures of allelic PCR products or genomic templates; an uneven mixture of dominant and trace alleles, by 9:1 and 8:1:1 mixtures. For 5:5, two homoduplex bands and two heteroduplex bands of equal intensity were generated. For 9:1, the trace homoduplex disappears whereas the two heteroduplexes are easily visible. For 8:1:1, four heteroduplexes and one homoduplex were observed; homoduplexes and heteroduplexes formed from the trace alleles were not visible. These experiments demonstrate that PCR can amplify mixed allelic templates in direct proportion to the stoichiometric fraction of each template. Trace species are captured as heteroduplexes with the most abundant species and are clearly displaced on denaturing gradient gels from the dominant homoduplex species. Our analytical studies can be applied to analysis of sequence variants in DNA collected from cancerous or infected tissues.(ABSTRACT TRUNCATED AT 250 WORDS)


Cystic fibrosis genotyping by direct PCR analysis of Guthrie blood spots. In the United States the most common cystic fibrosis (CF) alleles known are F508, G551D, G542X, R553X, and N1303K. These mutations comprise approximately 85% of U.S. CF alleles, and their detection along with analysis of XV-2C and KM-19 restriction fragment length polymorphisms (RFLPs) can enable the determination of CF status. To facilitate studies for determining CF carrier status, we developed methods to detect each of these mutations and RFLPs by direct PCR amplification of dried blood spots collected on newborn screening (Guthrie) cards. Following collection, samples were protected from contamination by individual plastic bags. One-mm2 segments of filter paper were added directly to 100-microliters PCR reactions containing 1/16 mM spermidine. Three initial cycles at 96 degrees C, then 55 degrees C, for 3 min were performed to free DNA and minimize inhibition by other related materials. Next, 1 unit of Taq polymerase was added and a 2-min extension was carried out at 72 degrees C, followed by 33 amplification cycles using denaturing, annealing, and extension temperatures and times optimal for each primer set. Then, 35 microliters of each reaction was run on 8% acrylamide gels directly or 1% agarose gels following digestion; genotypes were inferred by ethidium bromide staining of gels. Guthrie blood spots of 250 CF probands and their parents were screened and the frequencies of all five mutations as well as the XV-2C KM-19 RFLP haplotypes were determined.(ABSTRACT TRUNCATED AT 250 WORDS)


Expression, purification, and characterization of Bacneu. A soluble protein tyrosine kinase domain encoded by the neu-oncogene. To further characterize the structure and regulation of the tyrosine kinase encoded by the rodent neu oncogene, its cytoplasmic tyrosine kinase domain has been expressed as a soluble protein, called Bacneu, in Sf9 insect cells, using the baculovirus expression system. Expression of Bacneu was detected by immunoblotting with anti p185neu antisera and in vitro autophosphorylation analysis as early as 24 h postinfection. Maximal expression was observed at 48 h postinfection. The soluble kinase was purified to near homogeneity by sequential chromatography on DEAE-Sepharose, phosphocellulose, poly-L-lysine, and Sephacryl 300, yielding 0.55 mg Bacneu per L of Sf9 cells (4% yield). The kinase is more active in the presence of Mn2+ compared to Mg2+ ions. The specific activity of the kinase using poly(Glu4Tyr1) as a substrate is 179 nmol/min/mg. Maximal incorporation of 1.4 mol of phosphate per mol of enzyme by autophosphorylation was found to increase the activity of the enzyme 1.5- to twofold. These results indicate that the Bacneu kinase is activated by phosphorylation. Therefore, it will be a useful reagent for characterizing the effects that phosphorylation by other cellular kinases and dephosphorylation by phosphatases have on its activity.


Glutamate-induced inhibition of paired pulse facilitation of monosynaptic excitatory post-synaptic potentials in frog spinal motoneurons. To evaluate actions of glutamate on excitatory synaptic transmission in the central nervous system, we examined glutamate-induced changes in the paired pulse facilitation of monosynaptic excitatory post-synaptic potentials evoked by stimulation of the lateral column fibers (LC-EPSPs) on lumbar motoneurons in the frog spinal cord. Glutamate (1 mM) depolarized motoneurons both in the presence and absence of Mg2+. In most cells perfused with Mg(2+)-free or high Ca(2+)-Mg2+ solutions, the glutamate potential was accompanied by a reduction in peak amplitude of EPSPs, although the degree of change varied with the cells. Glutamate enhanced the EPSP amplitude in a few cells with Mg(2+)-free and high Ca(2+)-Mg2+ solutions, and in most cells with high Mg2+ medium. In 3/5 cells tested, the paired pulse facilitation of EPSPs was reduced by glutamate when the EPSP amplitude either increased or decreased. NMDA (50 microM), kainate (50-100 microM), quisqualate (5-50 microM) and L-2-amino-4-phosphonobutyrate (L-AP4, 1 mM) also decreased the facilitation in about half of the cells tested. The glutamate-induced decrease in the facilitation was observed in both the presence and absence of Mg2+ and was not affected by the concomitant application of glutamate and antagonists for non-NMDA or NMDA receptors, such as 6-cyano-7-nitro-quinoxalinediones (CNQX, 60 microM) or 2-amino-5-phosphonovalerate (APV, 250 microM). Glutamate reduced the facilitation of excitatory post-synaptic currents (EPSCs) recorded at a constant membrane potential under voltage clamp, when the EPSC amplitude either increased or decreased and when the input conductance either increased or decreased.(ABSTRACT TRUNCATED AT 250 WORDS)


Interactive role of L-glutamate and vasopressin, at the level of the PAG area, for cardiovascular tone and stereotyped behaviour. The periaqueductal gray (PAG) area may modulate cardiovascular functions and trigger several stereotyped behavioural responses through a mechanism mediated by the interaction of L-glutamate with arginine vasopressin (AVP). Moreover, only the NMDA- but not the non-NMDA-glutamergic subtype receptors might participate in the control of these neurovegetative functions also modifying the homeostasis of the hypothalamic-neurohypophysis system. This latter effect may be due to the tight connections between the PAG area neurons to the more cephalic nuclei within the brainstem.


Bacterial conjugation: everybody's doin' it. Recent evidence suggests that bacterial conjugation is ubiquitous in the bacterial world and that DNA transfer between different genera-kingdoms is possible. It has also been demonstrated that many bacterial gene transfer systems resemble each other at the molecular level and that others are a blend of two or more of these systems. Thus, in the absence of a sexual cycle, bacterial conjugation, along with bacteriophages, transposons, and natural transformation systems, forms a potent force for gene dissemination and repair in the eubacteria and simple eukaryotes.


Effect of D1 and D2 agonists in primates withdrawn from long-term treatment with haloperidol: the potential role of dopamine D1 receptors in dyskinesia. The effects of dopamine (DA) D1 and D2 receptor agonists were evaluated in eight Cebus apella monkeys. The monkeys had previously received haloperidol for 2 years, and five of the monkeys had developed mild oral dyskinesia. SKF 81297 (a full D1 agonist) induced marked oral hyperkinesia, consisting of tongue protrusions and licking or chewing movements, most pronounced in the monkeys with pre-existing oral dyskinesia. SKF 38393 and SKF 75670 (partial D1 agonists) also induced some oral dyskinesia, but to a lesser extent than SKF 81297, and with few licking movements. The partial D1 agonists, but not the full agonist, induced sedation. All of the D1 agonists induced grooming behavior, the full D1 agonist to the greatest extent. In the case of SKF 81297, the grooming was closely associated with the licking behavior. Quinpirole (a selective D2 agonist) and apomorphine (a mixed D1/D2 agonist) induced a hyperactive syndrome (nonoral stereotypy with rapid repetitive movements and increased arousal and locomotor activity). Quinpirole induced no grooming behavior and reduced pre-existing oral movements. The data indicate behavioral differences between D1 and D2 receptors and suggest that D1 receptors may be involved in the pathophysiology of some forms of dyskinesia syndromes.


Effects of alpidem in anxious elderly outpatients: a double-blind, placebo-controlled trial. The efficacy and safety of alpidem, a new anxiolytic imidazopyridine, were compared with those of placebo in anxious elderly patients (65-80 years) by means of a randomized, double-blind, parallel group study. Following a 7-day "placebo run-in," 40 anxious patients were randomized to receive either alpidem or placebo. Daily doses ranging from 75 to 150 mg (25-50 mg t.i.d.) were administered for 3 weeks. Hamilton Rating Scale for Anxiety (HRSA), State Trait Anxiety Inventory (STAI-X1), Visual Analogue Scale (VAS), and Clinical Global Impression (CGI) were used on days 0, 3, 7, 14, and 21 for assessing efficacy. Psychomotor and mnesic performances were evaluated at the same time by means of the Digit Symbol Substitution Test (DSST), the Grünberger's test for fine motor coordination, and the Hawie's test for immediate memory. Possible adverse events were also recorded during the five visits. The anxiolytic efficacy of alpidem was significantly (p < 0.01) superior to that of placebo in all the rating scales adopted. The anxiolytic action was clearly evident from day 7. For most of the patients the active dose was 25 mg t.i.d. No relevant adverse effects were observed in both groups. No impairment of psychomotor and mnesic performances could be observed in the alpidem group. Alpidem is a new interesting anxiolytic drug for anxious elderly patients because it appears remarkably safe and, at effective doses, it does not impair psychomotor performances and cognitive functions.


Prevention of myocardial infarction by nitroglycerin plus intravenous beta-blockers. Seven patients with ongoing angina with ST-segment elevation, refractory to oral nifedipine and intravenous nitroglycerin, were treated by adding intravenous beta-blockers. Chest pain resolved in all of them in a few minutes, and myocardial infarction did not develop in 5 patients. We recommend this approach for patients to whom thrombolytics are contraindicated or have been very recently administered, although further investigation is needed to extend its application more widely. It does not preclude the use of other therapies, if considered necessary.


[Alternative therapy in the acute phase of endogenous depression--antiepileptics and new benzodiazepines and their comparison]. The average therapeutic effect of classical tricyclic antidepressants and antidepressants of the second and third generation varies between 60-70%. Moreover, tricycklic antidepressants are associated with undesirable effects mostly anticholinergic and "cardiotoxic", which may be a contraindication in particular in patients with multiple diseases and in older age groups. The authors compared two new alternative therapeutic procedures, represented by new highly effective benzodiazepines and anticonvulsants, as regards the therapeutic effect and side-effects. From this comparison ensues that their general therapeutic effect is on average comparable with the effect of tricyclic antidepressants, however, in the spectrum of their undesirable symptoms the anticholinergic effects are absent. Recent benzodiazepines have a better effect on depressions where anxiety and agitation are in the foreground. Their effect on depressive core symptoms is more marked, as compared with anticonvulsants. The most frequent side-effects are fatigue, somnolence and vertigo. Anticonvulsants have a balanced effect in different syndromological forms of depression. Undesirable effects are mostly gastrointestinal. In the authors' trials it did not lead to hypomania.


Role of estrogen receptor variants in the development of hormone resistance in breast cancer. Recent evidence suggests that the progression to hormone resistance in some breast tumors is due to mutations in the estrogen receptor (ER). Various types of ER variants have been found in breast cancer biopsies and breast cancer cell lines. The ER variants include dominant-positive receptors that are transcriptionally active in the absence of estrogen, and dominant-negative receptors that are themselves transcriptionally inactive but prevent the action of the normal receptor. The mechanisms by which these variants cause loss of hormonal control is becoming clear. ER variants may be prognostic factors for breast cancer. By modifying the action of ER variants, it should be possible to develop new strategies for treatment of malignant breast disease.


Pulmonary pharmacology in pregnancy.

Pulmonary pharmacology in pregnancy. The pharmacologic treatment of any pregnant patient with a respiratory illness should occur only after careful consideration of the effects of altered maternal physiology and the potential effects on the developing fetus. Pregnancy changes gastrointestinal absorption of drugs, creates an increased volume of distribution, decreases protein binding, and increases renal excretion and hepatic metabolism of most drugs. These changes may necessitate variation in dose or dosing interval of drugs administered during pregnancy. Before administration of a therapeutic agent, the available animal and human data summarizing fetal effects should be reviewed. Most of the bronchodilators and more common antibiotics used for respiratory infections have been used without adverse fetal effects. Important exceptions do exist and certain agents should be avoided. When the patient expresses concern over a pharmacologic or radiologic exposure during pregnancy, a detailed history of the type, timing, and duration of exposure and genetic consultation should be obtained. Finally, drug treatment of serious illness in the pregnant patient should not be withheld unless the risk of the agent in question clearly outweighs the fetal and maternal risk of untreated disease.


Acute respiratory failure in pregnancy.

Acute respiratory failure in pregnancy. Acute respiratory failure in pregnancy is an important cause of maternal and fetal morbidity and mortality. Causes include: ARDS, venous air embolism, beta-adrenergic tocolytic therapy, asthma, thromboembolic disease, pneumothorax, and pneumomediastinum. The most common predisposing diseases for ARDS complicating pregnancy are sepsis, pneumonia, aspiration of gastric contents, and amniotic fluid embolism. Knowledge of normal maternal-fetal physiology and determinants of fetal oxygen delivery (uterine blood flow, placental transfer, fetal circulation) can help sustain normal fetal development, usually without compromising maternal care. The increased microvascular permeability seen in ARDS is likely mediated by neutrophils, proinflammatory mediators (e.g., tumor necrosis factor, interleukin-1, arachidonic acid metabolites) and activation of the complement cascade. Treatment of respiratory failure in pregnancy is largely supportive, including mechanical ventilation, hemodynamic support, nutrition, and prophylaxis against thromboembolism. No specific therapy has as yet been proven effective for ARDS, other than treating the underlying cause. Respiratory failure from status asthmaticus is treated with vigorous bronchodilator therapy, high-dose glucocorticosteroids, magnesium sulfate, and careful ventilator management. Occasionally, more experimental therapies (e.g., isoproterenol infusion, halothane anesthesia) are indicated. Certain strategies can help prevent respiratory failure from aspiration of gastric contents, beta-adrenergic tocolytic therapy, and thromboembolic disease.


Glutamine and glucose metabolism in bovine blood lymphocytes. 1. Glutamine and glucose metabolism was studied in bovine blood lymphocytes incubated at 37 degrees C in the presence of Krebs-Ringer bicarbonate buffer (pH 7.4) containing 1 mM [U-14C]glutamine and 5 mM [U-14C]glucose, respectively. 2. The major metabolic products from glutamine were ammonia, glutamate, and to a lesser extent, aspartate and CO2. Glucose was metabolized mainly to lactate and, to a lesser extent, pyruvate and CO2. These findings indicate incomplete oxidation of glutamine and glucose carbons in bovine blood lymphocytes. 3. Glucose provided three-fold greater amounts of energy to bovine blood lymphocytes than did glutamine on the basis of their measured end-products. Glycolysis accounted for 50% of glucose-derived ATP production. 4. Our findings suggest similar metabolic patterns of glutamine and glucose in lymphocytes between ruminants and non-ruminant species (e.g. rats). However, in contrast to rat peripheral lymphocytes, glucose, rather than glutamine, was a major energy substrate for bovine blood lymphocytes.


Restriction fragment length polymorphisms near the islet amyloid polypeptide gene in Japanese subjects. Two restriction fragment length polymorphisms (RFLPs) near the human islet amyloid polypeptide (IAPP) gene were examined in 50 Japanese patients with non-insulin-independent diabetes mellitus (NIDDM) and 54 non-diabetic controls. RFLPs were identified with the enzymes PvuII (A1 = 21 kb and A2 = 18 kb) and BglII (B1 = 9 kb and B2 = 7 kb). These RFLPs were in complete linkage disequilibrium with A1 which was in disequilibrium with B2, as was A2 with B1. Since these two RFLPs map to different locations in the 5'-flanking region of the IAPP gene, they are most likely due to changes in the sequence of the sites recognized by PvuII and BglII rather than to an insertion/deletion-type DNA polymorphism. There were no differences in the genotypic or allelic frequencies of these RFLPs between Japanese subjects with NIDDM and non-diabetic controls implying that these RFLPs do not play a major role in the development of NIDDM in this population.


Non-suppressed thyroidal radioactive iodine uptake (RAIU) in thyrotoxic phase in a case of subacute thyroiditis with thyroid-stimulating antibodies (TSAb). In this paper, we report a 49-year-old female with subacute thyroiditis who had thyroid-stimulating antibodies (TSAb) and thyroid-stimulation-blocking antibodies (TSBAb) in serum. Although she was in the thyrotoxic phase and TSH was suppressed in May, 1990, her radioactive iodine uptake (RAIU) was not suppressed (35.5%) and a thyroid scan disclosed a diffuse goiter with no defect. Serum assays revealed the presence of TSAb, but TSBAb were negative. In August, 1990, the right lobe became undetectable by thyroid scan when the RAIU was 20.7% with the TSH level remaining suppressed. At that time, TSAb were negative, while TSBAb were positive. When the RAIU was 31.1% in October, 1990, both thyroid lobes became visible and the TSH level was normalized. TSBAb became negative, and although TSAb reappeared it later became undetectable. These results indicate that the changes in the patient's thyroid scan and RAIU were attributable to the presence of TSAb.


Simultaneous measurement of changes in the membrane potential and the intracellular Ca2+ concentration caused by somatostatin in human GH-producing pituitary tumor cells. Changes in the membrane potential and the intracellular Ca2+ concentration ([Ca2+]i) caused by somatostatin (SRIF) were simultaneously measured in human GH-producing pituitary tumor cells, by means of the nystatin-perforated whole cell clamp technique and Fura-2 AM. An application of 10(-8) M SRIF hyperpolarized the membrane and arrested Ca(2+)-dependent spontaneous action potentials. [Ca2+]i concurrently decreased during membrane hyperpolarization. When the membrane potential was clamped below the threshold for voltage-gated Ca2+ channels, [Ca2+]i decreased and SRIF did not further reduce [Ca2+]i. In cells which did not show spontaneous action potentials, SRIF hyperpolarized the membrane but it affected [Ca2+]i little. From these results it was concluded that the reduction in [Ca2+]i caused by SRIF was ascribed to the decrease in Ca2+ influx through voltage-gated channels during membrane hyperpolarization. The effect of SRIF on the voltage-gated Ca2+ channel current was also examined under the perforated whole cell clamp. SRIF (10(-8) M) inhibited the Ca2+ channel current to 80.8 +/- 15.4% (n = 5) of the control. Because SRIF-induced inhibition of the voltage-gated Ca2+ channel current was not prominent, it was considered that membrane hyperpolarization is the major cause of the reduction in [Ca2+]i in human GH-producing cells.


Aortic input impedance in mild to moderate chronic congestive heart failure: lack of interrelation with neurohormonal activation. Patients with chronic congestive heart failure (CHF) have activated neurohormonal systems, which may induce vasoconstriction. In addition, the arterial wall sodium content may increase and could have direct trophic effects on vascular smooth muscle cells. These mechanisms might elevate left ventricular (LV) pulsatile load. We measured aortic input impedance to find out the LV pulsatile load and neurohormonal activation in 20 patients with mild to moderate chronic CHF. Cardiac index (2.2 +/- 0.3 l.min-1 x m-2) and LV ejection fraction (38 +/- 4%) were reduced, pulmonary wedge pressure was elevated (21 +/- 2 mmHg). Plasma norepinephrine levels (462 +/- 62 pg.ml-1), plasma renin concentration (12 +/- 4 ng.AI.ml-1h-1), plasma levels of atrial natriuretic peptide (408 +/- 64 pg.ml-1) and, to a slight degree, plasma arginine vasopressin (1.1 +/- 0.3 pg.ml-1) were increased. Characteristic impedance, Zc, was within the normal range (80 +/- 6 dyn.s.cm-5) and showed no significant correlation with the degree of neurohormonal activation (r-values: -0.05 to -0.35). Our data demonstrate that in early stages of CHF stimulation of the neurohormonal systems does not significantly elevate LV pulsatile load; therefore there is no substantial alteration in the physical properties of the great arteries.


Impact of primary surgical approach in the management of the impalpable testis. There is no univoque opinion about the place of preoperative studies in non-palpable testes. During a 6.5-year period, we operated on 296 impalpable testes in prepubertal boys. A combined inguinal-abdominal approach was used in all cases verifying the eventual abdominal testis and its exact vascular anatomy before any manipulation of the cord was undertaken. Forty-five testes (15.2%) were canalicular, 142 (48%) were abdominal, 5 (1.7%) dysgenetic and 104 (34.1%) absent (agenesis or vanishing testis). Of the abdominal testes, 122 underwent a standard orchidopexy in dartos pouch, 11 a staged repair, 8 a Fowler-Stephens operation and 1 orchiectomy. All means of investigation for impalpable testes are either unreliable, too expensive or too invasive for routine use, and in most cases, a surgical exploration has to be performed anyway. The primary surgical approach has the most favorable cost/benefit ratio, being diagnostic and therapeutic at one time. Provided the exploration is performed correctly, all the advantages of previous laparoscopy can be achieved with surgery alone.


Correlation between the expression of P-glycoprotein and multidrug-resistant phenotype in transitional cell carcinoma of the urinary tract. The expression of a multidrug-resistant (MDR) gene product, P-glycoprotein, was examined immunohistochemically in 41 transitional cell carcinomas (TCCs) of the urinary tract. In 23 of these, chemosensitivity to adriamycin (ADM) and vinblastine (VBL) was also assessed by a microtiter succinate dehydrogenase inhibition test and the correlation between the expression of P-glycoprotein and MDR phenotype was investigated. P-glycoprotein was detected in 13 (72.2%) of the 18 untreated TCCs of the upper urinary tract (UUT), 6 (31.6%) of the 19 untreated TCCs of the bladder, and all of the 4 TCCs treated with M-VAC chemotherapy, respectively. Fourteen (87.5%) of the 16 TCCs with a positive expression of P-glycoprotein were resistant to ADM and VBL, whereas all of the 4 TCCs sensitive to both drugs were negative in the expression of P-glycoprotein. The succinate dehydrogenase activity of TCCs with a positive expression of P-glycoprotein was significantly higher than that of TCCs with a negative expression of P-glycoprotein (P < 0.05). Thus, there was a good correlation between the expression of P-glycoprotein and MDR phenotype in the chemosensitivity test. These results suggest that intrinsic MDR exists in some TCCs of the urinary tract, particularly UUT, and that the immunohistochemical investigation of P-glycoprotein may be useful for predicting the MDR phenotype in TCCs of the urinary tract.


Early diagnosis of the multiple endocrine neoplasia type 2 syndrome: consensus statement. European Community Concerted Action: Medullary Thyroid Carcinoma. The diagnosis of medullary thyroid carcinoma by biochemical and genetic testing is possible in families with multiple endocrine neoplasia type 2. At an early stage total thyroidectomy usually cures the patient. As the clinical penetrance of the autosomal dominant, transmitted, multiple endocrine neoplasia type 2 gene is not complete, family screening is indicated for every new patient who presents with apparently sporadic medullary thyroid carcinoma. Problems related to a screening programme and early diagnosis have led the members of the European Community Concerted Action: Medullary Thyroid Carcinoma group to formulate a consensus on biochemical and genetic screening. For biochemical screening, measurement of basal and pentagastrin and/or calcium stimulated serum levels of calcitonin by radioimmunoassay are essential starting at the age of three and continuing annually until 35 years of age. Furthermore, annual screening for pheochromocytoma by measuring the urinary excretion of catecholamines and for hyperparathyroidism by serum calcium determination is indicated. Genetic screening using linked markers can be done with a 95% accuracy in informative families when DNA is available from at least two family members proven to be affected. Biochemical screening can thus be reserved for gene carriers, while those at low risk can be reassured. Combined biochemical and genetic screening for multiple endocrine neoplasia type 2 is important and effective for the cure of medullary thyroid carcinoma.


Noradrenergic mediation of spinal antinociception by 5-hydroxytryptamine: characterization of receptor subtypes. The present study examined the involvement of spinal noradrenergic mechanisms in spinal antinociception by the 5-hydroxy-tryptamine (5-HT) receptor-selective agonists CGS 12066B (5-HT1B; 7-trifluoromethyl-4(4-methyl-1-piperazinyl)-pyrrolo[1,2-a]quinoxaline), TFMPP (5-HT1C; M-trifluoromethylphenyl-piperazine) and DOI (5-HT2; 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane) using the rat hot plate test. Effects of alpha-adrenoreceptor antagonists (phentolamine, yohimbine), the adrenergic neurotoxin 6-hydroxydopamine, and the selective noradrenergic uptake blocker desipramine were determined. CGS 12066B, TFMPP and DOI produced dose-related antinociception. The antinociceptive effect of each agent was reduced by pretreatment with both phentolamine and yohimbine (15-60 micrograms). Pretreatment with 6-hydroxydopamine (100 micrograms, intrathecal) for 7-10 days, which reduced spinal cord levels of noradrenaline by 87%, inhibited the action of TFMPP (and 5-HT), but not CGS 12066B or DOI. Pretreatment with desipramine (25 mg/kg, systemic) potentiated the action of TFMPP but not CGS 12066B or DOI (or 2-methyl-5-HT). These results suggest that antinociception by TFMPP is dependent on release of endogenous noradrenaline from the spinal cord, while that produced by CGS 12066B and DOI is not. As TFMPP exhibits a close similarity to 5-HT in these experiments, the 5-HT receptor subtype being activated to induce noradrenaline release may either be a 5-HT1C or a 5-HT1S subtype. Other mechanisms account for the observed blockade of the action of CGS 12066B and DOI by alpha-adrenoreceptor antagonists.