Potential of 13C and 15N labeling for studying protein-protein interactions using Fourier transform infrared spectroscopy. In this study, we examine the interaction between two bacterial proteins, namely HPr and IIAmtl of the Escherichia coli phosphoenolpyruvate-dependent phosphotransferase system, using FTIR spectroscopy. In an interaction involving a 1:1 molar ratio of these two proteins, when they are unlabeled, the overlap of absorbance of the amide I band arising from the peptide group vibrations of the two proteins is such that it is not possible to determine the contribution which each protein makes to the absorbance. Uniform 15N labeling has little effect on the frequency of the amide I band although there is a significant shift of the amide II band. However, we show that uniform (90%) 13C labeling produces a large shift of bands associated with the carbonyl moiety, especially the amide I band. This opens up windows in different regions of the infrared spectrum. Thus, when the same mixture of the two bacterial proteins is made where one of the proteins is uniformly 13C-labeled (in our case HPr), the amide I maxima of this protein shifts by approximately 45 cm-1 toward lower frequency and reveals the previously overlapped amide I band of the unlabeled IIAmtl. This application of 13C labeling shows the potential of studying protein-protein interactions using FTIR spectroscopy. With thoughtful selection of systems and labeling strategies, numerous studies with proteins should be possible. These could include, among others, enzyme-substrate and protein-ligand interactions.


Identification in bovine liver plasma membranes of a Gq-activatable phosphoinositide phospholipase C. Phosphoinositide phospholipase C (PLC) activity extracted from bovine liver plasma membranes with sodium cholate was stimulated by GTP gamma S-activated G alpha q/G alpha 11, whereas the enzyme from liver cytosol was not. The membrane-associated PLC was subjected to chromatography on heparin-Sepharose, Q Sepharose, and S300HR, enabling the isolation of the G-protein stimulated activity and its resolution from PLC-gamma and PLC-delta. Following gel filtration, two proteins of 150 and 140 kDa were found to correspond to the activatable enzyme. These proteins were identified immunologically as members of the PLC-beta family and were completely resolved by chromatography on TSK Phenyl 5PW. The 150-kDa enzyme was markedly responsive to GTP gamma S-activated alpha-subunits of G alpha q/G alpha 11 or to purified Gq/G11 in the presence of GTP gamma S. The response of this PLC was of much greater magnitude than that of the 140-kDa enzyme. The partially purified 150-kDa enzyme showed specificity for PtdIns(4,5)P2 and PtdIns4P as compared to PtdIns and had an absolute dependence upon Ca2+. These characteristics were similar to those of the brain PLC-beta 1. The immunological and biochemical properties of the 150-kDa membrane-associated enzyme are consistent with its being the PLC-beta isozyme that is involved in receptor-G-protein-mediated generation of inositol 1,4,5-triphosphate in liver.


Structure of the glycan chain from the surface layer glycoprotein of Clostridium thermohydrosulfuricum L77-66. The thermophilic eubacterium Clostridium thermohydrosulfuricum L77-66 is covered by a crystalline surface layer composed of identical glycoprotein subunits which are arranged in a hexagonal lattice with centre-to-centre spacings of approx. 14.3 nm. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of cell wall preparations showed the presence of several broadened, carbohydrate-containing bands in a molecular mass range of 90 to 200 kDa. A total carbohydrate content of approx. 14% was determined in the purified surface layer glycoprotein. Chemical deglycosylation of this material by trifluoromethanesulfonic acid resulted in the disappearance of the complex banding pattern. Only a single band with a molecular mass of 82 kDa remained visible upon Coomassie staining. After proteolytic digestion of the surface layer glycoprotein a single glycopeptide fraction with an apparent molecular mass of approx. 25 kDa was obtained by gel filtration. Composition analysis, methylation, periodate oxidation and a combination of homonuclear and 1H-detected heteronuclear shift-correlated nuclear magnetic resonance experiments established the following structure for the glycan chain of the surface layer glycoprotein.


Non-redox protein interactions in the thioredoxin activation of chloroplast enzymes. Thioredoxin derivatives lacking SH groups such as S,S'-dicarboxymethyl-, dicarboxamidomethyl-thioredoxin and cysteine----serine mutant protein are capable of activating chloroplast NADP malate dehydrogenase and fructose-bisphosphatase when added to enzyme assays together with suboptimal amounts of native thioredoxin. The modified thioredoxins alone are inactive. These findings indicate that protein-protein interactions play a significant role in addition to disulfide/thiol exchange reactions in the light-driven regulation of plant enzymes by the various plant thioredoxins.


Biochemical characterization and substrate specificity of rat prostate kallikrein (S3): comparison with tissue kallikrein, tonin and T-kininogenase. A tissue kallikrein-like enzyme encoded by S3 mRNA was purified to homogeneity from rat prostate gland. The apparent molecular mass of the prostate enzyme is 32 kDa as determined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The intact 32 kDa enzyme is split into two bands of lower molecular mass, 18 and 14 kDa, under reducing conditions on SDS-PAGE. NH2-terminal amino acid sequence analyses of the intact enzyme and heavy and light chains revealed the identity to the translated sequence of a prostate kallikrein cDNA (S3). Isoelectric focusing indicated that the prostate enzyme is a basic protein with pI of 7.30-7.45. Specific activities of the prostate kallikrein toward angiotensin I, angiotensinogen and rat low M(r) kininogen as well as tripeptide chromogenic substrates were compared with those of tissue kallikrein, tonin and T-kininogenase. The kinin-releasing activity is inhibited by leupeptin, antipain, benzamidine and soybean trypsin inhibitor. A sensitive and specific radioimmunoassay for the rat prostate kallikrein shows that the immunoreactive kallikrein levels in prostate and submandibular gland were 23.78 +/- 2.62 micrograms/mg protein (n = 5) and 12.29 +/- 2.25 micrograms/mg protein (n = 5), respectively. The results indicate that the prostate kallikrein S3 is expressed at high levels in both prostate and submandibular glands.


Effects of sphingosine, albumin and unsaturated fatty acids on the activation and translocation of phosphatidate phosphohydrolases in rat hepatocytes. The activities of two phosphatidate phosphohydrolases were measured in cultured rat hepatocytes incubated with 0.1 mM albumin. The activity, which is inhibited by N-ethylmaleimide (PAP-1) is located in the cytosolic and membrane fractions. PAP-1 activity is stimulated by Mg2+ and it can be translocated from the cytosol to the membranes by relatively low (0.5-1 mM) concentrations of fatty acids. In addition, higher concentrations (1-3 mM) of fatty acids cause an increase in the total PAP-1 activity. Translocation of PAP-1 activity in the hepatocytes is preferentially promoted by unsaturated fatty acids (C18:1, C18:2, C18:3, C20:4 and C20:5), rather than by saturated acids (C14:0, C16:0, C18:0). Increasing the extracellular concentration of albumin from 30 microM to 1 mM displaces PAP-1 activity from the membrane fraction. Sphingosine, but not staurosporine, can inhibit the redistribution of PAP-1 activity induced by oleate. The amphiphilic amines, sphingosine, chlorpromazine and propranolol, also decrease membrane-bound PAP-1 activity in the absence of fatty acids, but they do not alter, significantly, the activity of the cytosolic PAP-1. In the presence of 1 mM oleate, sphingosine, chlorpromazine and propranolol decrease the translocation of PAP-1 from the cytosol to the membranes. The phosphohydrolase activity, which is insensitive to N-ethylmaleimide (PAP-2), is specifically located in the plasma membrane (Jamal, Z., Martin, A., Gomez-Muñoz, A. and Brindley, D.N. (1991) J. Biol. Chem. 266, 2988-2996) and it is not stimulated by Mg2+. Saturated fatty acids, albumin, sphingosine and propranolol have no significant effects on PAP-2 activity. However, chlorpromazine decreases PAP-2 activity by about 14%. Linolenate, arachidonate and eicosapentaenoate at 1 mM also produced small (7-10%) decreases in PAP-2 activity. It is proposed that both PAP-1 and PAP-2 activities may be involved in signal transduction, although the main function of PAP-1 seems to be involved in the synthesis of glycerolipids.


Cloning, nucleotide sequence and expression in Escherichia coli of a lipase gene from Bacillus subtilis 168. The gene coding for an extracellular lipase of Bacillus subtilis 168 was cloned and found to be expressed in Escherichia coli. Enzyme activity measurements showed no fatty acid chain length preference. A set of Tn5 insertions which inactivate the gene were localized and used to initiate its sequencing. The nucleotide sequence was determined on two independent clones expressed in E. coli. In one of these clones, the sequence revealed a frameshift, due to the presence of an additional adenine in the N-terminal region, which caused the interruption of the open reading frame, probably allowing translation to initiate at a second ATG codon. The sequence of the wild-type lip gene from B. subtilis was confirmed on the chromosomal fragment amplified by polymerase chain reaction (PCR). When compared to other lipases sequenced to date, the enzyme described here lacks the conserved pentapeptide Gly-X-Ser-X-Gly supposed to be essential for catalysis. However, alignments of several microbial lipase sequences suggest that the pentapeptide Ala-X-Ser-X-Gly present in the lipase B. subtilis may function as the catalytic site. Homologies were found in the N-terminal protein region with lipases from different Pseudomonas species. The predicted M(r) and isoelectric point for the mature protein are 19,348 and 9.7 respectively.


Primaquine-induced superoxide production by beta-thalassemic red blood cells. Primaquine, a prooxidant antimalarial drug, incubated with human red blood cells (RBC) induced marked superoxide generation in the cells as detected by exogenous cytochrome c reduction. In the presence of primaquine, beta-thalassemic RBC produced significantly more superoxide than normal RBC, thus reflecting the vulnerability of beta-thalassemic cells to oxidative stress.


Drug prevention programs can work: research findings. This paper reports findings of a subset of 91 programs, which included drug use measures, from the data base previously reported in the author's meta-analysis of 143 adolescent drug prevention programs. Treatment components of strategies successful in decreasing drug use by adolescents are discussed with regard to both the developmental stages of adolescents and the current etiology of drug abuse. Meta-analysis is briefly discussed. The focus is on issues rather than the actual research. Questions for future programming address theoretical assumptions and practical issues. Is attitude change a prerequisite for decreased drug use? This meta-analysis questions the validity of using knowledge and attitude measures as the only outcome measures. Successful program strategies require innovative planning and close attention to implementation factors. Answers to implementation questions require continued quality research. Implications for future planning may lie in the public policy arena.


Binding characteristics of delta opioid receptors in different regions of the brain of young and old male rats as studied with the highly selective ligand [D-Pen2-D-Pen5] enkephalin. The present experiments were performed to study the binding characteristics of delta opioid receptors in membrane preparations obtained from the brain of adult male rats, and to analyze whether aging modifies these binding parameters. The binding characteristics of delta opioid receptors were evaluated on membrane preparations derived from dissected brain regions (hypothalamus, amygdala, mesencephalon, corpus striatum, hippocampus, thalamus, frontal poles, anterior and posterior cortex) collected from male rats of 3 and 24 months of age; the highly selective ligand 3H-[D-Pen2-D-Pen5] enkephalin (3H-DPDPE) was used. The results obtained in young rats show that the distribution of delta opioid receptors is different in the various brain areas examined; these receptors appear to be maximally concentrated in the frontal poles, anterior and posterior cortex; lower concentrations were found in the other structures considered. Kd (dissociation constant) for the delta sites was found very similar in all areas. The distribution of delta opioid receptors in the brain of 24-month-old rats was similar to that observed in young animals; this result was surprising in view of the fact that aging modifies the number of other types of brain opioid receptors (mu and kappa).


A general diffusion model for analyzing the efficacy of synaptic input to threshold neurons. We describe a general diffusion model for analyzing the efficacy of individual synaptic inputs to threshold neurons. A formal expression is obtained for the system propagator which, when given an arbitrary initial state for the cell, yields the conditional probability distribution for the state at all later times. The propagator for a cell with a finite threshold is written as a series expansion, such that each term in the series depends only on the infinite threshold propagator, which in the diffusion limit reduces to a Gaussian form. This procedure admits a graphical representation in terms of an infinite sequence of diagrams. To connect the theory to experiment, we construct an analytical expression for the primary correlation kernel (PCK) which profiles the change in the instantaneous firing rate produced by a single postsynaptic potential (PSP). Explicit solutions are obtained in the diffusion limit to first order in perturbation theory. Our approximate expression resembles the PCK obtained by computer simulation, with the accuracy depending strongly on the mode of firing. The theory is most accurate when the synaptic input drives the membrane potential to a mean level more than one standard deviation below the firing threshold, making such cells highly sensitive to synchronous synaptic input.


The electrical coupling of two simple oscillators: load and acceleration effects. We consider two electrically coupled oscillators described by modified Fitzhugh-Nagumo equations. We study the relative influence of the individual cellular characteristics and the electrical coupling on the behavior of the coupled system. We show that, for similar oscillators, the load effect of the slow oscillator increases with the coupling strength. We prove that an asymmetry between the uncoupled bursters can accelerate the system with respect to the free cells, this effect depending on the characteristics of the coupling.


The relationship between the Gabor elementary function and a stochastic model of the inter-spike interval distribution in the responses of visual cortex neurons. In a previously reported study (Berger et al. 1990) we analyzed distributions of interspike intervals recorded extracellularly from cat visual cortex under four stimulus conditions. Stimuli were gratings differing in orientation and spatial frequency. The probability density function of first passage time for a random walk with drift process, which is defined by its barrier height and drift coefficient, was used to characterize the generating process of axonal discharge under resting and stimulus conditions. Drift coefficient and barrier height were derived from the sample mean and standard deviation of the measured inter-spike intervals. For cells with simple receptive fields, variations in spatial frequency produced changes only in drift coefficient. Variations in barrier height were produced only by changes in orientation of the stimulus. Currently, the method used to analyze these data was implemented in a simulation which displayed the relationship between the interval distribution of impulses, the random walk which represents the time series characteristic of the spike train model and the Gabor filter function which represents the geometry of the receptive field process.


Neurokinin concentrations in cerebrospinal fluid. A preliminary study in Parkinson's disease. Immunoreactive neurokinin A was measured in the cerebrospinal fluid of twelve patients with Parkinson's disease and eleven normal subjects, using a sensitive and precise extraction/concentration radioimmunoassay method. The mean value obtained in Parkinson's disease patients (13.2 +/- 4.6 pmol/l) was lower than that of the controls (17.4 +/- 5.9). The tendency toward a significant decrease (p = 0.085) found in this preliminary study could indicate that neurokinin A containing neurons are involved in the pathophysiology of Parkinson's disease. In addition, the establishment of reference values for neurokinin A in cerebrospinal fluid may provide a basis for further studies of this neuropeptide in neurological disorders.


Reduced activity of Na(+)-K+ ATPase of pancreatic islets in chronic renal failure: role of secondary hyperparathyroidism. The activity of Na(+)-K+ ATPase of pancreatic islets modulates their insulin secretion. The study presented here examined the activity of this enzyme in pancreatic islets of chronic renal failure (CRF) rats in an effort to further delineate the mechanisms of impaired insulin secretion in CRF. The Vmax of Na(+)-K+ ATPase, but not its Km, and the ATP content are significantly reduced in islets of CRF rats that have elevated levels of parathyroid hormone (PTH). These derangements are prevented by prior parathyroidectomy of CRF rats (low blood levels of PTH) or by their treatment with the calcium channel blocker verapamil; these latter rats have sustained elevation of blood levels of PTH. The data indicate that the chronic excess blood levels of PTH in CRF initiates events (augmented entry of calcium) that lead to the reduction in ATP content and in Vmax of Na(+)-K+ ATPase of pancreatic islets. Reducing the blood levels of PTH by parathyroidectomy or blocking the action of PTH on calcium entry into cells by verapamil prevents these derangements. The results suggest that chronic inhibition of Na(+)-K+ ATPase may participate in the processes underlying the impaired insulin secretion in CRF.


The postdialytic plasma cyclic guanosine 3':5'-monophosphate level as a measure of fluid overload in chronic hemodialysis. The postdialytic plasma level of cGMP, a marker for the release of atrial natriuretic peptide (ANP) in humans, is closely related to hypervolemia in chronic hemodialysis patients. In order to test the practicability of routine postdialysis cGMP determination for the detection of fluid overload, ANP and cGMP levels in the total hemodialysis population of 81 patients were measured with blood samples drawn immediately after hemodialysis. Twenty-three patients had a cGMP level of more than 20 pmol/mL. In 13 of these, pulmonary congestion was present on the chest roentgenogram. Two of these patients refused a gradual reduction of their dry body weight. In the remaining 21 patients, the weight reduction was associated with a decrease in cGMP levels in all cases and with a decrease in ANP levels in all but two cases. Fourteen of the 21 patients reached a cGMP level below 20 pmol/mL after weight reduction, and at that time, none of these showed signs of pulmonary congestion on chest x-ray. All seven patients, whose cGMP levels remained above 20 pmol/mL despite the reduction, had documented heart disease with impairment of left ventricular function. These results suggest that the plasma cGMP level after hemodialysis is more apt for the determination of dry body weight than is ANP or a chest roentgenogram.


Interleukin-1 and tumor necrosis factor: effector cytokines in autoimmune diseases. Autoimmune diseases have been studied from the perspective of an abnormal immune response in genetically vulnerable hosts. Although the immune response is responsible for the initiation of autoimmune diseases, the effectors of the disease process likely involves cytokines such as interleukin-1 (IL-1) and tumor necrosis factor (TNF). These polypeptides induce a wide variety of inflammatory events which contribute to the destruction of tissue and tissue remodeling in several autoimmune diseases. Blocking IL-1 with its naturally occurring receptor antagonist, the IL-1 receptor antagonist reduces the severity of disease in animal models of inflammation and autoimmune processes. Clinical studies with the IL-1 receptor antagonist will define the role for this cytokine in the pathogenesis of autoimmune diseases such as arthritis, inflammatory bowel disease, type I diabetes and vasculitis.


A monoclonal antibody to a novel surface antigen, MKW, blocks the antiproliferative and differentiation effects of granulocyte-macrophagecolony-stimulating factor and vitamin D3. A monoclonal antibody (MoAb) recognizes a novel 52-Kd cell protein (MKW) that is expressed on cells of the normal myelocytic and monocytic lineage, a subset of B cells, and the U937 cell line. Using the U937 cell line as a model, the MoAb (anti-MKW) was examined for its ability to inhibit the effects of differentiation-inducing factors. In the U937 cell line, recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) inhibits cell proliferation, 12-O-tetradecanoylphorbol-13-acetate (TPA) inhibits proliferation and induces the early differentiation antigen CD11b, and vitamin D3 inhibits proliferation and induces both CD11b and the late differentiation antigen CD14. The antiproliferative and differentiation effects of rhGM-CSF and vitamin D3 on U937 cells were inhibited by the anti-MKW MoAb. Similar effects were seen when anti-MKW antibody was added 30 minutes before or 2 hours after rhGM-CSF or vitamin D3, suggesting that its effects are not mediated by blocking or binding to the receptors for these growth factors. The anti-MKW MoAb had no effect on TPA-induced differentiation in U937 cells, indicating that TPA exerts its effects through a pathway different from rhGM-CSF and vitamin D3. These results suggest that the MKW antigen is important in controlling the proliferation and differentiation of monocytic cells.


Apoptotic cell death of primed CD45RO+ T lymphocytes in Epstein-Barr virus-induced infectious mononucleosis. The expansion of activated T cells, characterized by the expression of CD45RO as well as HLA-DR antigens, is a central feature in acute infectious mononucleosis (IM) induced by primary infection of Epstein-Barr virus (EBV). However, the fate of these activated T cells in this disease is not clearly understood. We found that, on simple culture, a large proportion of T cells isolated from acute IM patients died rapidly, but only a few T cells from normal individuals did. Morphologic observations and DNA fragmentation analysis showed that the loss of viability of IM T cells after incubation was mediated by apoptosis. IM T cells undergoing apoptosis resided exclusively in the CD45RO+ populations of both CD4+ and CD8+ T cells, most of which were shown to coexpress apoptosis-related Fas antigen. Some cytokines such as interleukin-2 (IL-2), IL-5, and IL-6 could rescue IM T cells from apoptotic cell death. The results seemed to imply that most of primed (CD45RO+) T cells in acute IM might be subject to apoptotic cell death, possibly when leaving from the local sites actively producing certain soluble factors required for their survival. Our studies suggest the programmed cell death of peripheral mature T cells as a mechanism of antigen-driven selection.


Distribution and phenotype of Epstein-Barr virus-harboring cells in Hodgkin's disease. Distribution and phenotype of Epstein-Barr virus (EBV)-harboring cells were determined in Hodgkin's disease (HD) biopsies by in situ hybridization with [35S]-labeled RNA probes specific for the small EBV-encoded nuclear RNAs, EBER1 and EBER2, in some instances preceded by immunohistology for CD20, CD30, CD45RO, and CD68 antigens, the T-cell receptor beta-chain, and latent membrane antigen (LMP) of EBV. Twenty-three of 46 HD cases displayed EBER transcripts in all Hodgkin and Reed-Sternberg (H-RS) cells, and 18 of these cases showed LMP expression exclusively in neoplastic cells. EBER+ small reactive cells were present in 39 cases in low numbers, and in three cases in abundance. Thus, presence of H-RS cells with or without LMP expression was not accompanied by an unrestricted proliferation of reactive EBER+/LMP- lymphoid cells in the majority of HD patients. Simultaneous in situ hybridization with [35S]-labeled immunoglobulin light chain (IgLC) gene probes and nonisotopically labeled EBER probe showed a phenotype of mature B lymphocytes and a polyclonal composition for a large proportion of the EBER+ small cells. However, in contrast to noninfected cells, CD20 expression was not detectable in many of these cells, which may indicate downregulation of certain differentiation antigens in latently EBV-infected small lymphoid cells in vivo.


Molecular monitoring of the myl/retinoic acid receptor-alpha fusion gene in acute promyelocytic leukemia by polymerase chain reaction. The acute promyelocytic leukemia (APL) t(15;17) translocation generates a myl/retinoic acid receptor-alpha (RAR-alpha) chimeric gene that is transcribed as a fusion myl/RAR-alpha messenger RNA. Using primer sets derived from RAR-alpha and myl cDNAs, we were able to amplify the breakpoint sites of the fusion transcripts of all 35 APL RNA samples by reverse polymerase chain reaction (PCR) and nested primer approach of two rounds of amplification. DNA fragments of different size were obtained according to the chromosome 15 breakpoints (intron 3-bcr 3; exon 6-bcr 2; and intron 6-bcr 1). bcr 1 and bcr 3 represent the regions of the myl locus most frequently involved among APL (48.5 and 34.2 of cases, respectively); bcr 3 constitutes 62.5% of cases among M3V as compared with 25.9% of M3 cases. The feasibility of monitoring the APL clone by PCR analysis in five APL patients who received different treatment (chemotherapy, all-trans-retinoic acid or bone marrow transplantation) was evaluated. In five of nine bone marrow samples of patients in complete remission, t(15;17)-positive cells could be detected by PCR analysis. We conclude that PCR amplification of the myl/RAR-alpha junctions represents the easiest and rapid method for diagnosis and monitoring of the APL clone.


Interleukin-6 production in high-grade B lymphomas: correlation with the presence of malignant immunoblasts in acquired immunodeficiency syndrome and in human immunodeficiency virus-seronegative patients. The mechanisms leading to malignant cell proliferation may differ between the different histologic forms of high-grade non-Hodgkin's lymphomas. To analyze the potential role of interleukin-6 (IL-6) as a growth factor for lymphomatous cells in these different forms, the in situ production of this cytokine was analyzed in lymphomatous samples taken from 24 patients, 18 of whom were human immunodeficiency virus (HIV) infected. Eleven Burkitt's lymphomas (BLs), seven diffuse large-cell lymphomas, and six immunoblastic lymphomas were studied. In situ hybridization experiments showed that the IL-6 gene was expressed in all tissues. The number of IL-6 gene-expressing cells was 7 times higher in the non-BLs than in the BLs, and it was 17 times higher than that of 14 control lymph nodes displaying a benign follicular hyperplasia. Analysis of individual cases indicated that the level of IL-6 gene expression was strongly correlated with the presence of immunoblasts within the malignant clone. In contrast, this level was not correlated with the presence of Epstein-Barr virus genome in the lymphoma or with the HIV status of patients. Immunohistochemical studies with an anti-IL-6 monoclonal antibody showed that IL-6 was produced in non-BLs, but not in BLs. In the former, IL-6 mainly originated from reactive, nonmalignant cells. Immunohistochemical analyses of non-BLs also showed that malignant cells produced the 80-Kd chain of the IL-6 receptor. Taken together, these results suggest that IL-6 may act as a growth factor in some forms of high-grade B lymphomas. The presence of immunoblasts may be an indicator of such forms.


Decreased pyrimidine nucleoside monophosphate kinase activity in sickle erythrocytes. We have previously shown that physiologic concentrations of hemin cause marked inhibition of several red blood cell (RBC) enzymes. Because endogenous heme content is elevated in sickle RBCs, we have examined the activity of hemin-sensitive enzymes in these RBCs. One of the hemin-sensitive enzymes, pyrimidine nucleoside monophosphate kinase (PNMK), was shown to have decreased activity in sickle RBCs relative to RBCs of equivalent cell age. The other hemin-sensitive enzymes, including adenylate kinase (AK), pyrimidine 5'-nucleotidase (P5N), 6-phosphogluconate dehydrogenase (6PGD), and aldolase, had activities that were appropriate for cell age. We have also examined the affinity of the hemin-sensitive enzymes to hemin. Using two different methods, PNMK was shown to have the highest binding affinity to hemin. The exquisite sensitivity of PNMK to inhibition by hemin, coupled with the enzyme's high affinity to hemin, may account for the decrease in PNMK activity and the lack of significant decrease in the other hemin-sensitive enzymes in sickle RBCs. These results suggest that the increased endogenous heme content in sickle RBCs may be responsible for the decrease in PNMK activity. Whether the increased endogenous heme content of sickle RBCs can cause hemolysis indirectly by inhibiting RBC enzymes remains to be determined.


Human papillomavirus in breast cancer.

Human papillomavirus in breast cancer. Histological sections from paraffin-embedded breast carcinoma and axillary lymph nodes were examined for the presence of human papillomaviruses by two different techniques: the polymerase chain reaction (PCR) and the in situ hybridization with biotin-labelled probes. By PCR we detected HPV 16 DNA sequences in 29.4% of breast tumours and in some metastatic lymph nodes, though we were unable to identify any HPV DNA sequences by in situ hybridization. These results suggest that HPV's could play a role in the genesis of breast neoplasia.


The human glutamate receptor cDNA GluR1: cloning, sequencing, expression and localization to chromosome 5. The rat glutamate receptor is a 907 amino acid transmembrane protein. Using the rat GluR1 cDNA as a probe, we have isolated cDNA clones from a human hippocampal cDNA library. Sequence of a full length cDNA clone revealed 98.2% and 89.4% identity to the rat sequence at the amino acid and nucleotide levels respectively. The human cDNA clone detected an RNA transcript in human cerebral cortex, hippocampus and cerebellum, similar to that seen in rat. In situ hybridization experiments showed that human GluR1 mRNA is present in granule and pyramidal cells in the hippocampal formation and that there is no apparent difference of distribution between control patient and patient with Alzheimer's disease. Dot blot analysis of flow-sorted human chromosomes showed that the GluR1 gene maps to chromosome 5.


Fission yeast sts1+ gene encodes a protein similar to the chicken lamin B receptor and is implicated in pleiotropic drug-sensitivity, divalent cation-sensitivity, and osmoregulation. The Schizosaccharomyces pombe sts1+ gene, identified by supersensitive mutations to a protein kinase inhibitor, staurosporine, was isolated by complementation by the use of a fission yeast genomic library. Nucleotide sequencing shows that the sts1+ gene encodes a 453 amino acid putative membrane-associated protein that is significantly similar (26% identity) to the chicken lamin B receptor. It is also highly related (53% identity) to a budding yeast ORF, YGL022. These three proteins contain a similar hydrophobicity pattern consisting of eight or nine putative transmembrane domains. By gene disruption we demonstrate that the sts1+ gene is not essential for viability. These disruptants exhibit pleiotropic defects, such as cold-sensitivity for growth and at the permissive temperature, a supersensitivity to divalent cations and several unrelated drugs including staurosporine, caffeine, chloramphenicol, sorbitol, and SDS. Disruption of the sts1+ gene does not lead to a sensitivity to thiabendazole or hydroxyurea.


Molecular cloning and developmental expression of the catalytic and 65-kDa regulatory subunits of protein phosphatase 2A in Drosophila. cDNA clones encoding the catalytic subunit and the 65-kDa regulatory subunit of protein phosphatase 2A (PR65) from Drosophila melanogaster have been isolated by homology screening with the corresponding human cDNAs. The Drosophila clones were used to analyze the spatial and temporal expression of the transcripts encoding these two proteins. The Drosophila PR65 cDNA clones contained an open reading frame of 1773 nucleotides encoding a protein of 65.5 kDa. The predicted amino acid sequence showed 75 and 71% identity to the human PR65 alpha and beta isoforms, respectively. As previously reported for the mammalian PR65 isoforms, Drosophila PR65 is composed of 15 imperfect repeating units of approximately 39 amino acids. The residues contributing to this repeat structure show also the highest sequence conservation between species, indicating a functional importance for these repeats. The gene encoding Drosophila PR65 was located at 29B1,2 on the second chromosome. A major transcript of 2.8 kilobase (kb) encoding the PR65 subunit and two transcripts of 1.6 and 2.5 kb encoding the catalytic subunit could be detected throughout Drosophila development. All of these mRNAs were most abundant during early embryogenesis and were expressed at lower levels in larvae and adult flies. In situ hybridization of different developmental stages showed a colocalization of the PR65 and catalytic subunit transcripts. The mRNA expression is high in the nurse cells and oocytes, consistent with a high equally distributed expression in early embryos. In later embryonal development, the expression remains high in the nervous system and the gonads but the overall transcript levels decrease. In third instar larvae, high levels of mRNA could be observed in brain, imaginal discs, and in salivary glands. These results indicate that protein phosphatase 2A transcript levels change during development in a tissue and in a time-specific manner.


Internal calcium release and activation of sea urchin eggs by cGMP are independent of the phosphoinositide signaling pathway. We show that microinjecting cyclic GMP (cGMP) into unfertilized sea urchin eggs activates them by stimulating a rise in the intracellular free calcium ion concentration ([Ca2+]i). The increase in [Ca2+]i is similar in both magnitude and duration to the transient that activates the egg at fertilization. It is due to mobilization of calcium from intracellular stores but is not prevented by the inositol trisphosphate (InsP3) antagonist heparin. Furthermore, cGMP does not stimulate the eggs Na+/H+ antiport when the [Ca2+]i transient is blocked by the calcium chelator bis-(O-aminophenoxy)-N,N,N',N'-tetraacetic acid (BAPTA), suggesting that cGMP does not activate eggs by interacting with the their phosphoinositide signaling pathway. However, the [Ca2+]i increase and activation are prevented in eggs in which the InsP3-sensitive calcium stores have been emptied by the prior microinjection of the InsP3 analogue inositol 1,4,5-trisphosphorothioate. These data indicate that cGMP activates eggs by stimulating the release of calcium from an InsP3-sensitive calcium store via a novel, though unidentified, route independent of the InsP3 receptor.


Lack of correlation between hormonal blood levels and endometrial maturation in agonadal women with repeat implantation failure following embryo transfer from donated eggs. Five women with ovarian failure who repeatedly failed to conceive following embryo transfer from donated eggs underwent endometrial development investigation. One endometrial biopsy was obtained on cycle days 19, 21, and 23 during three consecutive artificially induced cycles. All five patients had only early secretory changes on days 19 and 21. Histological evaluation on cycle day 23 revealed various developmental stages: two women had "in-phase" endometrium, two patients had adequately developed stroma but significantly retarded glandular maturation, and one women showed no progress. The histological findings were conclusive for a significant maturation delay and an impaired endometrial receptivity. There was a lack of correlation between the peripheral hormonal blood levels and the endometrial maturation.


[Treatment of infantile hepatitis syndrome with injection of yin zhi huang]. The infantile hepatitis syndrome is defined as a liver disease which occurs during the infantile period with the clinical manifestation of jaundice, hepatomegaly and hepatic dysfunction. The cause of disease is complex and the treatment is difficult. From March 1987 to June 1988, 55 patients (treatment group) were treated with injection of yin zhi huang, and 15 cases (control group) with injection of Inosine, vitamin C and glucose. The course of treatment lasted 15 days. At the beginning and the end of the treatment, serum bilirubin level, sALT level, duodenal juice bilirubin level, liver size and T cell subsets in peripheral blood had been detected separately. The results showed that among the treatment group, serum bilirubin level was significantly lower at the end of the treatment than that of the control group. The sALT, and hepatomegaly were also reduced or lessened, while the duodenal juice bilirubin level was increased. But there was no difference in T cell subsets between the two groups.


[A clinical study on allicin in the prevention of thrush in newborn infants]. This article deals with the 0.06/1000 allicin and 2.5% sodium bicarbonate in order to look for effective drugs in preventing thrush. The results revealed: (1) The incidence of the disease of the two drugs in the less dangerous group was significantly decreased compared with that of the control (P less than 0.01); (2) In the more dangerous group, the incidence of the disease of the allium group was more significantly decreased than that of the control, but no significant decrease in sodium bicarbonate was observed. The two drugs are both effective in preventing thrush and the allium is more effective.


Thymus ontogeny in frogs: T-cell renewal at metamorphosis. Metamorphosis in amphibians presents a unique problem for the developing immune system. Because tadpoles are free-living, they need an immune system to protect against potential pathogens. However, at metamorphosis, they acquire a variety of new adult-specific molecules to which the tadpole immune system must become tolerant. We hypothesized that Xenopus laevis tadpoles may avoid potentially destructive antiself responses by largely discarding the larval immune system at metamorphosis and acquiring a new one. By implanting triploid (3N) thymuses into diploid (2N) hosts, we examined the influx and expansion of host T-cell precursors in the donor thymus of normally metamorphosing and metamorphosis-inhibited frogs. We observed that donor thymocytes are replaced by host-derived cells during metamorphosis, but inhibition of metamorphosis does not prevent this exchange of cells. The implanted thymuses export T cells to the spleen. This donor-derived pool of cells declines after metamorphosis in normally developing frogs but is retained to a greater extent if metamorphosis is inhibited. These studies confirm previous observations of a metamorphosis-associated wave of expansion of T cells and demonstrate that it is not dependent on the relatively high concentrations of thyroid hormones required for metamorphosis. Although some larval T cells persist through metamorphosis, others may be destroyed or the larval population is significantly diluted by the expanding adult population.


TCR V alpha- and V beta-gene segment use in T-cell subcultures derived from a type-III bare lymphocyte syndrome patient deficient in MHC class-II expression. Previously, we and others have shown that MHC class-II deficient humans have greatly reduced numbers of CD4+CD8- peripheral T cells. These type-III Bare Lymphocyte Syndrome patients lack MHC class-II and have an impaired MHC class-I antigen expression. In this study, we analyzed the impact of the MHC class-II deficient environment on the TCR V-gene segment usage in this reduced CD4+CD8- T-cell subset. For these studies, we employed TcR V-region-specific monoclonal antibodies (mAbs) and a semiquantitative PCR technique with V alpha and V beta amplimers, specific for each of the most known V alpha- and V beta-gene region families. The results of our studies demonstrate that some of the V alpha-gene segments are used less frequent in the CD4+CD8- T-cell subset of the patient, whereas the majority of the TCR V alpha- and V beta-gene segments investigated were used with similar frequencies in both subsets in the type-III Bare Lymphocyte Syndrome patient compared to healthy control family members. Interestingly, the frequency of TcR V alpha 12 transcripts was greatly diminished in the patient, both in the CD4+CD8- as well as in the CD4-CD8+ compartment, whereas this gene segment could easily be detected in the healthy family controls. On the basis of the results obtained in this study, it is concluded that within the reduced CD4+CD8- T-cell subset of this patient, most of the TCR V-gene segments tested for are employed. However, a skewing in the usage frequency of some of the V alpha-gene segments toward the CD4-CD8+ T-cell subset was noticeable in the MHC class-II deficient patient that differed from those observed in the healthy family controls.


The calcium second messenger system in bipolar disorders: data supporting new research directions. Studies of aberrations in second messenger function may help to elucidate some of the multiple complex neurobiological alterations in bipolar affective disorders. The phosphatidylinositol and calcium ion (Ca2+) second messengers are of particular interest because of evidence of hyperactivity of these signaling mechanisms in both mania and bipolar depression and of their normalization by lithium and other mood-stabilizing treatments. Because the intracellular Ca2+ signal has a biphasic action, a single aberration could explain diverse clinical manifestations of the same illness, and a single action on the messenger could explain the biphasic actions of many treatments for bipolar disorders.


Lifetime complications of drug use in intravenous drug users. Ninety-two intravenous drug users (IVDUs) were identified from a study of 1,640 relatives of treated alcoholics and felons. Nearly all of the IVDUs (90%) reported a lifetime history of some degree of physical, psychological, or social difficulty related to drug abuse, but only 35% had ever received treatment for drug abuse. Overall, compared to subjects with a history of substantial illicit drug use who had never injected, IVDUs were significantly more likely to report all drug-related problems assessed. Even in populations of drug users not selected through treatment, any history of intravenous (IV) drug use is strongly indicative of a lifetime occurrence of complications of drug use.


The relationship between craving, anxiety, and other symptoms in opioid withdrawal. Craving and anxiety share many similar attributes including comparable responses to pharmacological agents. To better characterize the relationship among craving, anxiety, and other opioid withdrawal signs and symptoms, we examined 32 opioid-dependent subjects undergoing clonidine-assisted opioid withdrawal. A 13-item analog scale measured subjective withdrawal symptoms. Statistical analyses of separate subscale items, using craving as the dependent variable, showed that the highest positive correlations occur for craving and anxiety. The high correlation between craving and anxiety suggests common mechanisms underlying both.


Social support and depression in men during alcoholism treatment. Psychiatric comorbidities have been shown to be important predictors of the outcome of alcoholism treatment. This study examines whether perceived lack of social support can be identified as an independent predictor of symptoms of depression experienced during alcoholism treatment over and above the effects of personality characteristics and the severity of alcohol and psychiatric history. We studied 189 alcoholic men in treatment at a rural midwestern Department of Veterans Affairs medical center. Multiple regression analyses found that reduced social support significantly predicted depression (measured by the Beck Depression Inventory) during treatment while controlling for personality characteristics and the alcoholism and psychiatric subscales of the Addiction Severity Index. Although self-esteem, neuroticism, and psychiatric severity also were significantly associated with depression in the hierarchical regression model, social support demonstrated the strongest unique contribution to depression of any of the predictors. These results suggest that social support has an independent association with depression and perhaps may play an important role in improving treatment compliance and the outcome of alcoholism treatment.


A survey of state Medicaid policies for coverage of screening mammography and Pap smear services. In the winter of 1990, we surveyed all states and the District of Columbia to ascertain Medicaid policies on screening mammography and Pap smear--two critical preventive cancer screens for women. Forty-four state Medicaid programs cover screening mammography and all 51 jurisdictions cover Pap smear services. However, the extent of coverage and reimbursement rates vary widely across states. Only a small minority of states reported age or frequency limits for screening; five of these are in conflict with nationally recommended guidelines.


Effects of camptothecin, an inhibitor of DNA topoisomerase I on ribosomal gene structure and function in TG cells. The effects of camptothecin treatment and topoisomerase I inhibition on ribosomal gene structure and function were investigated in TG cells, a human tumour cell line. 90- and 180-min treatments with 25 microM camptothecin resulted in an increased DNA fragmentation and decreased activity of topoisomerase I in cell extracts. After 180-min treatment, the incorporation of labelled uridine into total cell RNA was reduced to 39% and the ribosomal RNA synthesis to 10%, as compared to values of control cells. At the ultrastructural level, the nucleolar components appeared to be segregated; after selective DNA staining, with osmium-amine complex, a part of the nucleolar chromatin of treated cells showed the presence of thin, extended DNA filaments, superimposable to those present in control cells.


The non-peptide NK1 receptor antagonist, (+/-)-CP-96,345, produces antinociceptive and anti-oedema effects in the rat. The non-peptide NK1 receptor antagonist, (+/-)-CP-96,345, has been evaluated for antinociceptive activity in two well-characterized inflammatory pain models in the rat. (+/-)-CP-96,345 abolished carrageenin-induced mechanical hyperalgesia, significantly reduced carrageenin-induced paw oedema and attenuated the second phase of the formalin response. The results suggest that NK1 receptor activation occurs during the induction of inflammatory pain states in the rat.


Heterogeneity of endothelin-sarafotoxin receptors mediating contraction of pig coronary artery. The contractile effects of endothelin-1, endothelin-3, sarafotoxin 6b and sarafotoxin 6c were studied in endothelium-denuded rings of pig coronary artery. Endothelin-1, sarafotoxin 6b and sarafotoxin 6c produced monophasic concentration-response curves (mean EC50 values 6.7, 14.8 and 1.6 nM), whereas the concentration-response curve to endothelin-3 was biphasic (mean EC50 values 9.6 nM and 0.32 microM). The maximal effect of sarafotoxin 6c was about one third of that reached by the other peptides. The higher sensitivity component of the curve to endothelin-3 was abolished in the presence of sarafotoxin 6c (0.3 microM), while the EC50 value for the other component remained unchanged. Sarafotoxin 6c (0.3 microM) failed to alter the EC50 values of endothelin-1 and sarafotoxin 6b. These data strongly suggest the presence of at least two endothelin-sarafotoxin receptors mediating contraction of pig coronary artery, one with the profile of the endothelin ETA receptor subtype, the other recognizing sarafotoxin 6c and endothelin-3, but not endothelin-1 and sarafotoxin 6b, being thus different from the ETB receptor subtype.


Cardiovascular actions of the kappa-agonist, U-50,488H, in the absence and presence of opioid receptor blockade. 1. The cardiovascular actions of U-50,488H, a kappa-receptor agonist, were studied in rat isolated perfused hearts, and in anaesthetized rats, over concentrations or doses generally above those required to produce kappa-receptor-mediated effects. 2. U-50,488H dose-dependently decreased left-ventricular peak systolic pressure and beating rate in vitro and reduced blood pressure and heart rate in vivo. 3. Over the concentration range of 1-30 microM in vitro, and the dose-range of 0.5-32 mumol kg-1 in vivo, U-50,488H prolonged the P-R, QRS and Q-T intervals of the ECG. 4. The effects of U-50,488H were not antagonized by an opioid receptor antagonist, naloxone (1 microM or 8 mumol kg-1). Similarly, the opioid receptor antagonist, MR 2266, at 8 mumol kg-1 did not significantly reduce the cardiovascular actions of U-50,488H in vivo. 5. The actions of U-50,488H on responses to electrical stimulation were also studied. Over the dose range of 0.5-32 mumol kg-1, U-50,488H altered thresholds and effective refractory period. It had a biphasic action on thresholds for induction of ventricular fibrillation. Thresholds were decreased at lower doses (0.5-4 mumol kg-1) but increased at higher doses (8-32 mumol kg-1). The effects of lower doses were blocked by naloxone. Effective refractory period and threshold pulse width only increased with dose. 6. In conclusion, U-50,488H at high concentration, had direct depressant actions on cardiac contractility, electrical excitability and the ECG. These depressant effects were not antagonized by the opioid receptor antagonists, naloxone and MR 2266, and probably do not involve opioid receptors. Furthermore, some of the observed effects were those expected to result from sodium channel blockade.


Studies on the mechanism of 5-HT1 receptor-induced smooth muscle contraction in dog saphenous vein. 1. We have investigated the mechanism of smooth muscle contraction evoked by activation of 5-HT1-like receptors in dog isolated saphenous vein. 2. In the presence of the 5-HT2 receptor antagonist, ritanserin (0.1 microM), concentration-effect curves (10 nM-300 microM) for 5-hydroxytryptamine (5-HT)-induced smooth muscle contraction were biphasic. This could be attributed to a direct action on 5-HT1-like receptors at low concentrations of 5-HT (10 nM-10 microM) and an indirect (through the release of noradrenaline from sympathetic neurones) activation of postjunctional alpha-adrenoceptors at higher 5-HT concentrations. In contrast, concentration-effect curves (100 nM-100 microM) for sumatriptan-induced contractions were not biphasic, and were due solely to activation of 5-HT1-like receptors. 3. Smooth muscle contractions evoked either by low concentrations of 5-HT or by sumatriptan were abolished by removal of extracellular calcium and were markedly inhibited, but not abolished, by the calcium channel blocker, verapamil (1-30 microM). In contrast, contractions evoked by high concentrations of 5-HT were markedly less sensitive to removal of extracellular calcium or to verapamil. 4. 5-HT and sumatriptan also inhibited (to a maximum of about 50%) prostaglandin E2 (PGE2, 5 microM)-stimulated adenosine 3':5'-cyclic monophosphate (cyclic AMP) formation. This effect was mimicked by the alpha 2-adrenoceptor agonist, azepexole (B-HT933) but not by the alpha 1-adrenoceptor agonist, methoxamine.5. In contrast to mediation of smooth muscle contraction, the 5-HT1-like receptor-mediated inhibition of PGE2-stimulated cyclic AMP formation evoked by 5-HT or sumatriptan was not attenuated by removal of extracellular calcium or by verapamil (1 microM).6. A directly-acting inhibitor of adenylyl cyclase, 2',3'-dideoxyadenosine (1 mM) inhibited PGE2-stimulated cyclic AMP formation but did not produce smooth muscle contraction.7. These results suggest that contractile responses of dog isolated saphenous vein arising through activation of 5-HT1-like receptors are associated with both an influx of extracellular calcium ions (to a large extent via voltage-dependent channels) and an inhibition of adenylyl cyclase. However, although these two responses are coupled to the same receptor, they appear to be independent.


Separation of putative alpha 1A- and alpha 1B-adrenoceptor mediated components in the tension response of the rat vas deferens to electrical field stimulation. 1. The effects of the putative alpha 1B-adrenoceptor antagonist, chloroethylchlonidine (CEC), on tension responses of the rat isolated whole vas deferens to single and multiple pulses of electrical field stimulation have been evaluated by use of a microcomputer system which enables the averaging of like-responses throughout their time course. 2. CEC (10(-7) to 3 x 10(-6) M) selectively and in a concentration-dependent manner blocked the noradrenergic component of the response to a single field stimulus in the absence or presence of nifedipine (10(-5) M, which blocked the purinergic but not the noradrenergic component of the response). The concentration-response curve of the vas to exogenously-applied noradrenaline (NA) was unaffected by CEC (10(-6) M) but was flattened by nifedipine (10(-5) M). 3. The tension response to 10 Hz trains of pulses was biphasic, with an early (less than 2 s) and a plateau (greater than 4 s) phase. We deduce from our pharmacological analysis that the early phase contains a putative alpha 1B-adrenoceptor component (susceptible to CEC or prazosin but not to nifedipine) and a P2-purinoceptor component (susceptible to suramin or nifedipine) whereas the plateau phase contains an alpha 1A-adrenoceptor component (susceptible to prazosin or nifedipine but not to CEC) and a P2-purinoceptor component (susceptible to suramin or nifedipine). 4. We suggest that the putative alpha 1B-adrenoceptors may be functionally confined to the synaptic region whereas the putative alpha 1A-adrenoceptors are excluded from this region. Trains of pulses would allow NA to accumulate and spill out beyond the synaptic region to reach and activate the putative alphalA-adrenoceptors.


Radiotherapy to the surviving kidney after unilateral nephrectomy in bilateral Wilms' tumour. Despite an improving overall survival rate for children with Wilms' tumour, four out of seven patients with bilateral tumours died in the period from 1952 to 1960 and five out of eight died in the period from 1971 to 1989, at St Bartholomew's Hospital and the Hospital for Sick Children. More aggressive chemotherapy with both adriamycin and actinomycin D and concern over young age being predisposed to late radiation morbidity prompted us to keep radiotherapy dose prescriptions to the surviving kidney below the quoted renal radiation tolerance dose equivalent. In three long-term survivors treated with daily fractions up to 167 cGy and total doses of 1000-1200 cGy, we have found renal function and growth to be within the "normal" range at follow-up and the patients to be normotensive 6-8 years later. As four of the eight patients reported here died from local disease progression within the kidney (albeit despite slightly larger dose prescriptions), we discuss the potential for larger total doses to be delivered in this situation.


Vasoactive intestinal polypeptide presynaptically enhances the synaptic transmission in cultured sympathetic neurons. We studied the effects of vasoactive intestinal polypeptide (VIP) on the cholinergic synaptic transmission that was developed between rat sympathetic neurons in culture. Electrophysiological examinations revealed that the amplitude of fast excitatory postsynaptic potential (fast EPSP) was increased by VIP (0.2-0.8 microM) reversibly and dose-dependently, whereas transient nicotinic depolarization evoked by pressure application of acetylcholine (ACh) was not affected by VIP. In most of the cells examined, VIP depolarized membrane potential by a few millivolts with concomitant increases in membrane conductance. Furthermore, the VIP-induced depolarization was suppressed by Co2+ but not by hexamethonium or atropine. Hence it is highly likely that the peptide augmented the amplitude of fast EPSPs by increasing ACh release from the presynaptic cell. These results demonstrate that VIP influences the presynaptic phase of cholinergic synaptic transmission between sympathetic neurons.


Age-related changes in binding to excitatory amino acid uptake sites in human cerebellum. In vitro autoradiography and test-tube assay of the sodium-dependent binding of D-[3H]aspartate were used to localize and quantify the uptake site for the excitatory amino acid neurotransmitters glutamate and aspartate in the cerebellar cortex of human cerebellar hemispheres. Autoradiograms revealed a pronounced heterogeneity in the distribution of D-[3H]aspartate binding in cortex from adult brains, with the highest binding density corresponding to the Purkinje cell layer, high binding in molecular layer and low binding in granule cell layer. In contrast, cerebellar cortex from infants at term (40 weeks gestation) had only low binding of the ligand in both the molecular and the Purkinje cell layers. Both methods employed for measuring D-[3H]aspartate binding showed that the number of binding sites in Purkinje and molecular layers increased rapidly from term to 20 weeks postnatal age and achieved levels higher than those found in adult cerebellum. It is concluded that a substantial increase in the numbers of glutamate/aspartate uptake sites takes place in the human cerebellum during the early postnatal period. It is deduced that the excess uptake sites are eliminated as the cerebellum matures.


Genetic differences in hypothalamic-pituitary-adrenal axis responsiveness to acute ethanol and acute ethanol withdrawal. There is a growing body of evidence suggesting that corticosteroids contribute to the increased neural excitability observed during ethanol withdrawal. In the present study, this was further investigated using mouse strains which differ in ethanol withdrawal severity. DBA/2 (DBA) mice were found to display more severe acute ethanol withdrawal seizures than C57BL/6 (C57) mice. Additionally, DBA mice showed a greater stress response than C57 mice, as measured by higher plasma concentrations of adrenocorticotropic hormone (ACTH) and corticosterone, to an acute dose of ethanol. Mimicking withdrawal plasma corticosterone levels by administering corticosterone to ethanol-naive mice resulted in increases in handling-induced convulsions in the range observed during withdrawal. There did not appear to be a strain difference in sensitivity to the excitatory effects of corticosterone. In summary, the greater stress response to ethanol by DBA mice may account, in part, for the more severe ethanol withdrawal syndrome of this strain.


Localization of cytochrome oxidase (COX) activity and COX mRNA in the hippocampus and entorhinal cortex of the monkey brain: correlation with specific neuronal pathways. Cytochrome oxidase (COX) activity and COX II mRNA expression were localized in the hippocampal formation and entorhinal cortex of the rhesus monkey brain by means of enzyme histochemistry and in situ hybridization, respectively. Within the hippocampal formation, the terminal field of the perforant pathway showed the highest levels of COX activity, whereas COX II mRNA was localized mainly in neuronal cell bodies. In the entorhinal cortex. COX II mRNA was detected in neuronal cell bodies of layers II and IV. These results indicate that the pattern of localization of COX and its mRNA in entorhinal cortex correlates with the input and output pathways of the hippocampus.


The long-term effect of nedocromil sodium on the maximal degree of airway narrowing to methacholine in atopic asthmatic subjects. Airway hyperresponsiveness in asthma is characterized by increased airway sensitivity and by excessive maximal airway narrowing. Long-term inhalation therapy with nedocromil sodium has been shown to reduce increased airway sensitivity in asthma. However, it is unknown whether it also attenuates excessive airway narrowing. We studied the long-term effects of nedocromil on the maximal degree of airway narrowing to methacholine. Twenty-seven atopic asthmatic adults (21-39 years), with a measurable maximal-response plateau on the dose-response curve (20-55% fall in FEV1), were randomly allocated into two parallel treatment groups. They received either inhaled nedocromil 4 mg q.i.d. or placebo, for 8 weeks following a 2 week baseline period. Every 2 weeks, complete dose-response curves to inhaled methacholine were obtained. The response was measured by FEV1 and by volume history standardized partial expiratory flow-volume curves (V40p). A maximal-response plateau was considered if three or more of the highest data points fell within a 5% response range, the maximal response being the average value on the plateau (MFEV1, MV40p). Airway sensitivity was defined as the provocative concentration of methacholine causing a 20% fall in FEV1 (PC20FEV1) or 40% fall in V40p (PC40 V40p). Twenty-four subjects completed the study. Baseline FEV1 or V40p did not change during either treatment (P greater than 0.07). There were no significant changes in MFEV1 or MV40p during treatment with nedocromil (P greater than 0.07). Neither were these changes significantly different between the two groups (P greater than 0.25).(ABSTRACT TRUNCATED AT 250 WORDS)


Plasma triglycerides: a cardiovascular risk factor? In a span of 3 decades, investigation as to whether elevated plasma triglyceride (TG) is associated with increased risk of coronary artery disease (CAD) produced results that have largely been inconsistent. Therefore, these investigations have been relegated to the background when compared to the significance give to cholesterol in the 80's. The first study to report an association between TG and myocardial infarction was in 1959. Since then, many other studies have been done starting from case control studies to prospective studies and then progressing to interventional studies. Many have found TG significant on a univariate but not on a multivariate analysis. Attempts have been made to analyse TG in diabetics, especially those with a normal cholesterol, in ischemic cerebrovascular disease (ICD), and in other disease states where atherosclerosis is involved in their pathogenesis. Of particular interest is that our studies in Taiwan have consistently showed that TG may indeed be an independent risk factor. Our subjects with ICD, CAD, and in different stages of hypertension, all had significant association with elevated TG. Statistical consider, i. e. discrepancy between univariate and a multivariate analysis, large variability of plasma TG distribution, as well as genetic susceptibility of certain individuals are also factors to be considered. No study to date has specifically been designed to evaluate TG lowering effects on primary prevention of CAD. Despite differences of opinion among the various authorities on this subject, I feel that the TG-CAD association should not be ignored and further intervention studies should be done.


Response of left ventricular ejection performance following balloon valvuloplasty in patients with mitral stenosis. In order to see if any change of left ventricular ejection performance (LEVP) would occur after successful balloon mitral valvuloplasty (BMV), a prospective study on echocardiography and calibrated carotid pulse tractings was performed two days before, two days and then six months after BMV in 24 patients with pure rheumatic mitral stenosis (MS). Echocardiographic parameters representing preload i.e. end-diastolic volume index (EDVI); afterload i.e. end-systolic wall stress (ESS), and the indices of LVEP i.e. ejection fraction (EF), fractional shortening, rate-corrected velocity of circumferential shortening (VCFc) and a ratio of ESS over end-systolic volume index (ESS/ESVI) were measured. A group of 66 normal subjects was used to establish the 95% confidence interval of the echo parameters. The incidence of depressed LVEP in MS patients was about 21% when measured by load-independent index i.e. ESS/ESVI. In the MS group, 7 out 24 (29%) patients achieved an increased cardiac index after BMV. Six (25%) MS patients showed an EF below 95% confidence interval of control (less than 55%) before BMV and, of these 6, only 3 patients increased their EDVI after the BMV. Those who showed an EF greater than or equal to 55% before BMV gave no change in their preload (EDVI) after the BMV. Only one MS patient showed high afterload (ESS) both before and after BMV. However, the ESS/ESVI showed insignificant improvement (p = 0.055) after BMV. It was therefore concluded that (1) a depressed LVEP is frequently seen in patients with MS; (2) LVEP can be improved by BMV in some (21%) patients with MS, and the improvement is not related to the change of preload or afterload but more likely secondary to an increased myocardial contractility.


External gastrointestinal fistula after the advent of total parental nutrition. From 1984 to 1988, 173 patients with 190 external gastrointestinal fistulae and given total parenteral nutrition (TPN) were reviewed. The advent of TPN has enabled the patients to be operated in a better condition under a well-planned procedure, and has therefore modified the principle of surgical intervention. However, no significant improvement has been noted in the mortality rate and spontaneous closure rate as compared with the years without TPN. The diseases have changed in pattern to afflict older population and increase the incidence of complicated fistula because of the more sophisticated surgery. It may also have something to do with delayed intervention of the intraabdominal abscess and delayed surgical closure of the fistula due to TPN administration. Early and aggressive treatment against intraabdominal infection, and early closure of the fistulae which fail to close spontaneously with 6-week conservative treatment after subsidence of infection, are advocated.


Radionuclide renal function study in various surgical treatments of upper urinary stones. We conducted a radionuclide renal function study using 131I-hippuran in 188 patients with upper urinary stones before and every 3 months to the maximum of 12 months after various surgical treatments. Of 28 patients with renal staghorn stone, 24 underwent anatrophic nephrolithotomy and 4 percutaneous nephrolithotomy (PCNL). Of 36 patients with renal pelvic stone, 15 received open pyelolithotomy, 16 received PCNL and 5 received extracorporeal shock wave lithotripsy (ESWL). Of 71 patients with renal calyceal stone, 48 underwent PCNL and 23 ESWL. Of 53 patients with ureteral stone, 41 received ureterolithotomy, 8 PCNL and 4 ureteroscopic extraction. The renal function study determined individual and total renal function in terms of effective renal plasma flow (ERPF) at unilateral and bilateral kidneys. The mean total ERPF decreased 3-6 months after anatrophic nephrolithotomy (377.4 +/- 121.8 vs. 308.8 +/- 105.4 ml/min) or PCNL (447.4 +/- 152.3 vs. 386.8 +/- 140.8 ml/min) for staghorn stone. The mean ERPF of the kidney on the operative side decreased significantly to an extent of 29.4% up to 6 months after anatrophic nephrolithotomy. The mean ERPF on operative side increased significantly to an extent of 18.3% up to 6 months after ureterolithotomy. In conclusion, among the upper urinary stones, the unilateral (operative side) and total bilateral renal function decreased 1, 3, and 6 months, respectively, after anatrophic nephrolithotomy. On the contrary, the renal function increased significantly on the operative side up to 6 months after open ureterolithotomy. There was no significant change of mean ERPF at the kidney on operative or non-operative side in other kinds of upper urinary stones and surgical treatment.


Predicting the probability of malignancy of the neck mass with logistic regression model: a statistical analysis of excisional biopsy of neck masses. One hundred and fifty-six patients with neck lesions were selected into this retrospective study between January, 1989 and December, 1990. All the patients visited OPD with the chief complaint of neck mass without other apparent symptoms and signs after initial work-up. They can be divided into 5 types according to the pathological reports: type a) metastatic lesion, type b) malignant lymphoma, type c) TB lymphadenitis, type d) miscellaneous benign lesion, and type e) inadequate specimen. They represent 24.4%, 12.8%, 9.6%, 52.6%, and 0.6% of the patients respectively. Type a) and type b) were classified as group of malignancy and the other three types were group of benignancy. Chi-square test and t-test were then used to evaluate the significant level of difference in each semiological factor between both groups. Several parameters were found to reach significant level, including tumor fixation, age of the patients, tenderness, location and size of the tumor, and history of cancer. Stepwise logistic regression was utilized to obtain a regression equation to predict the probability of malignancy in OPD patients with neck masses. The accuracy rate of prediction is 83.3%, if the cutpoint value is 0.5. A clinician can therefore avoid untimely excisional biopsy and delay in treatment planning.


Experience of six patients with Vibrio vulnificus septicemia. Vibrio vulnificus, a halophilic lactose fermenting vibrio, is a virulent pathogen for men and is frequently associated with overwhelming infections of areas other than the gastrointestinal tract. We encountered six cases of Vibrio vulnificus septicemia in Veterans General Hospital-Taipei over the past four years. All were admitted through the emergency room and presented with urgent conditions on arrival. The patients also demonstrated preexisting liver function impairment (either hepatic disease or chronic alcohol consumption). Five subjects had an apparent history of exposure to marine environments: one fisherman with pre-existing wounds and four others with previous consumption of poorly cooked seafood. Characteristic hemorrhagic bullous lesions were found in 5 cases. In all, 4 patients (67%) died with three of the cases within 24 hours of hospitalization. Misdiagnosis and delayed treatment were the most common causes. In conclusion, when patients present with sepsis and/or characteristic cutaneous lesions, particulously those with underlying liver disease and a history of marine exposure, clinicians should be alerted to this potentially fatal infection and should commence appropriate assessment and treatment immediately.


[Function endoscopic sinus surgery in children]. Sinusitis is commonly seen in children. There is evidence that both acute and chronic sinusitis in children is a dynamic, multifactorial disease. Generally most patients will be cured by medical treatment. Surgery is indicated when medical treatment fails. Functional endoscopic sinus surgery (FESS), the procedure of ostiomeatal reconstruction, had become a popular surgical treatment for sinus diseases in the world. We collected 22 children with chronic sinusitis who received FESS between June, 1990 and September, 1991. The children's age ranged from 6 to 16 years. Postoperative follow-up was from 3 to 18 months. All patients tolerated the procedure well, and there were no major complications. Synechia between the middle turbinate and lateral nasal wall occurred in 47% children. Fair results were obtained in 86% patients. Well-trained functional endoscopic sinus surgery is a safe procedure and benefits in treatment of pediatric sinusitis.


[Incidence of pulmonary aspiration with different kinds of artificial airways]. A patient with an artificial airway in place, pulmonary aspiration is a serious complication. A prospective study was conducted to investigate the incidence of pulmonary aspiration in patients with all kinds of modern artificial airways with high-volume low-pressure cuffs. In ICU and NSCU (neurosurgery care unit), 40 patients with a tracheostomy or endotracheal tube were included. Before study, cuff pressure was checked and remained at 25 cmH2 O if possible. Then, 0.2 ml of 1% methylene blue was applied on the tongue of each patient every 4 hours. Routine tube care was performed including frequent suction at least once an hour. Any evidence of the blue dye-marker obtained on suctioning was considered positive. The duration of study was 24 hours. Among these 40 patients, the procedure was performed in 50 episodes because some of the patients were evaluated with different kinds of artificial airways. Thirty episodes were evaluated with endotracheal tube, including 7 oral and 23 nasotracheal tubes. No episode of aspiration could be found (0/30). With tracheostomy tubes, 5 of 20 episodes showed positive result (5/20). The difference of incidence between these 2 groups was statistically significant (p less than 0.01). Some possible contributing factors were evaluated between positive and negative episodes, including age, sex, respiration rate, different brands of tracheostomy tubes, modes of ventilation, PEEP level, cuff pressure, nasogastric tubes, coma scale, posture, and ratio of tube and tracheal diameters in tracheostomy group.(ABSTRACT TRUNCATED AT 250 WORDS)


[Intracranial germ cell tumors].

[Intracranial germ cell tumors]. We report on 19 cases of intracranial germ cell tumors, including 12 male and 7 female patients; the average age was 16.5 +/- 4.9 years. These tumors included germinoma in 15 cases, yolk sac tumor in 3 cases, embryonal cell carcinoma in 1 case, and mixed tumor in 3 cases. Each of 5 cases had 2 to 3 different kinds of tumor cells, either in one tumor or in different tumors. The locations of tumors were pineal region in 13 cases, suprasellar region in 3 cases, basal ganglion-thalamus in 3 cases, and cerebral hemispheres in 4 cases. Six of them had 2 or more locations. One case had spinal seeding and extraneural metastasis to the liver, retroperitoneum and neck lymph nodes. Eighteen cases received CT scan and 6 cases received MRI examinations, all the tumors had good enhancement by iodium content contrast medium in CT and Gadolinium-DTPA in MRI. Alfa-fetoprotein was elevated in yolk sac tumor, AFT and HCG were elevated only slightly or remained normal in germinoma. Germinomas had good response to radiation therapy and chemotherapy, while nongerminoma had poor response and worse prognosis.


Myositis ossificans progressiva mimicking ankylosing spondylitis: a case report. Myositis ossificans progressiva is a rare disorder of young adults characterized by ossification of the connective tissue of the voluntary muscles and ligaments. Although it is trauma-related, up to 40-60% of these patients have no history of previous injury. A young female with marked kyphosis and ankylosis of the spine presented with a recent onset of a rapidly growing painful mass over the anterior aspect of her left shoulder. She received an excisional biopsy but recurrent ossification developed soon after. It then spread to the biceps muscle with subsequent contracture deformities of the shoulder and elbow joints. A plain radiogram of her spine revealed similar characteristics of ankylosing spondylitis. However, the final diagnosis was made by the pathognomonic ectopic ossification of muscles and para-articular soft tissue. Despite poor response of the established constracture, the painful mass did respond well to prednisolone treatment within 2 months, in terms of size and consistency.


[Squamous cell carcinoma of the gallbladder associated with adenocarcinoma in situ: a case report]. Squamous cell carcinoma of the gallbladder associated with adenocarcinoma in situ of the mucosal columnar epithelium is extremely rare. The case presented here is the first to be documented at the Veterans General Hospital-Taipei. In this case, dysplastic columnar epithelium was discovered alongside with squamous cell carcinoma and squamous metaplasia. Focal adenocarcinoma in situ was also identified. The significance of these associated lesions and the histogenesis of squamous metaplasia and squamous cell carcinoma in gallbladder are discussed.


Laser induced molar tooth pulp chamber temperature changes. Temperature changes in enamel tissue and the pulp chamber under the influence of a CO2 laser were measured by direct methods in vitro. X-ray diffraction analysis revealed alpha-Ca3(PO4), the high-temperature modification of enamel hydroxyapatite, thus indicating that the enamel melting temperature was above 1,000 degrees C in the interaction area of laser (continuous wave, 15 s exposure time, 1 mm spot size) and tissue. Powers of 0.5 and 1 W (continuous wave), 1.5 mm spot size, and 10 s exposure time vaporize and carbonize dentin tissue at the cavity bottom of class I preparation molars. The observed temperature rise of 4 degrees C indicates that thermal injury to the pulp tissue does not occur.


Receptor-mediated and absorptive endocytosis by male germ cells of different mammalian species. The routes for adsorptive and receptor-mediated endocytosis were studied in vivo after microinjection of tracers into the lumen of the seminiferous tubules, and in vitro in isolated germ cells of different mammals. Cationic ferritin was located on the plasma membrane, in vesicles, in tubules, in multivesicular bodies and in lysosome-like granules of mouse spermatocytes. In these cells the number of multivesicular bodies varied during spermatogenesis. Spermatids and to a lesser extent residual bodies also performed adsorptive endocytosis. In the rat and monkey (Macaca fascicularis) diferric transferrin was specifically taken up by germ cells via receptor-mediated endocytosis. The labelling was observed subsequently in membrane pits, vesicles, endosome-like bodies and pale multivesicular bodies. A progressive decrease in the frequency of the labelling of the germ cells by transferrin-gold particles was observed from spermatogonia to spermatocytes and to early spermatids, which could indicate that iron is particularly required by germ cells during the mitotic and meiotic processes. Adsorptive and receptor-mediated endocytosis therefore occurs in all classes of germ cells. These endocytic processes are most probably required for germ cell division, differentiation and metabolism.


Two distinct isoforms of sea urchin egg dynein. Extracts of unfertilized sea urchin eggs contain at least two isoforms of cytoplasmic dynein. One exhibits a weak affinity for microtubules and is primarily soluble. The other isoform, HMr-3, binds to microtubules in an ATP-sensitive manner, but is immunologically distinct from the soluble egg dynein (Porter et al.: Journal of Biological Chemistry 263:6759-6771, 1988). We have now further distinguished these egg dynein isoforms based on differences in NTPase activity. HMr-3 copurifies with NTPase activity, but it hydrolyzes CTP at 10 times the rate of ATP. The soluble egg dynein is similar to flagellar dynein in its nucleotide specificity; its MgCTPase activity is ca. 60% of its MgATPase activity. Non-ionic detergents and salt activate the MgATPase activities of both enzymes relative to their MgCTPase activities, but this effect is more pronounced for the soluble egg dynein than for HMr-3. Sucrose gradient-purified HMr-3 promotes an ATP-sensitive microtubule bundling, as seen with darkfield optics. We have also isolated a 20 S microtubule translocating activity by sucrose gradient fractionation of egg extracts, followed by microtubule affinity and ATP release. This 20 S fraction, which contains the HMr-3 isoform, induces a microtubule gliding activity that is distinct from kinesin. Our observations suggest that soluble dynein resembles axonemal dynein, but that HMr-2 is related to the dynein-like enzymes isolated from a variety of cell types and may represent the cytoplasmic dynein of sea urchin eggs.


[Heterogeneous carcinoma of the gallbladder with neuroendocrine differentiation]. A heterogenous mostly neuroendocrine small cell carcinoma was found in a gallbladder resected from a 75-year-old man suffering from CLL. It progressed along bile ducts into choledochus, into omentum and liver. Its histology was characterized by solid alveoli and small cell trabeculae with a high mitotic activity, dissociated infiltration in some parts and desmoplasia. Sometimes a typical adenocarcinomatous differentiation could be found out. Tumour cells produced immunohistological expression of EMA, CEA, NSE and CHG and had Grimelius silver impregnation positivity. Neuroendocrine gallbladder carcinomas used to be more aggressive than carcinomas of another type, can cause an endocrine syndrome and claim a special treatment.


Calculation of received dose intensity for combinations of drugs using small-cell lung carcinoma treatment regimens as examples. Programs are presented for the calculation of received dose intensity in combination chemotherapy regimens. These provide methods for determining the final dose intensity, the mean cumulative dose intensity together with its standard error, and other tabular and graphic summaries. Two ways of dividing patients into high and low received-dose-intensity groups are proposed. Methods are illustrated using data from Mid-Atlantic Oncology Program (MAOP) 2183, a phase III evaluation of a six-drug alternating combination vs a three-drug "standard" combination treatment for extensive small-cell lung cancer. Comparisons of received dose intensity with demographic and outcome variables are presented.


Mitomycin C plus vindesine or cisplatin plus epirubicin in previously treated patients with symptomatic advanced non-small-cell lung cancer. A total of 40 previously treated patients with symptomatic advanced non-small-cell lung cancer (NSCLC) were subjected to second-line chemotherapy with mitomycin C plus vindesine (MV) or cisplatin plus epirubicin (PE). The 12 patients treated with the MV regimen showed no objective response (OR) or symptom palliation. In the 28 patients who received the PE regimen, we obtained a 25% partial response rate, with amelioration of tumor-related symptoms occurring in 35.7% of cases and improvement in the performance status being noted in 25% of subjects. Both regimens were well tolerated. These data show that the administration of cisplatin-based second-line chemotherapy to patients with symptomatic advanced NSCLC may be useful.


Phospholipid N-methylation in diabetic erythrocytes: effects on membrane Na+, K+ ATPase activity. Phospholipid methylation was quantified in non-diabetic and streptozotocin diabetic rat erythrocytes. While the total mass of methylated lipids remained the same in both groups, the relative abundance of individual methylated lipid species differed significantly in diabetic erythrocytes. Moreover, incubation of erythrocytes membranes with S-adenosyl methionine, a substrate for methyl transferases, not only increased membrane lipid methylation but also decreased Na+, K+ ATPase activity significantly. These results suggest that phospholipid methylation may cause the observed depression of erythrocyte Na+, K+ ATPase activity in diabetes and could contribute to the altered rheology of erythrocytes in diabetes.


Developmental changes in long-opening behavior of L-type Ca2+ channels in embryonic chick heart cells. In the early (3-day) stage of development, long-lasting openings of the L-type Ca2+ channels (mode 2) occur in embryonic chick heart cells. Since mode-2 behavior is infrequently observed in adult heart cells of other species, in the present study, developmental change in behavior of the Ca2+ channel was examined in young (3-day) and old (17-day) embryonic chick heart cells. In the whole-cell voltage clamp, the L-type Ca2+ current carried by Ca2+ ions was smaller in amplitude and had a faster inactivation in 17-day cells than in 3-day cells. The peak current density was 8.1 +/- 0.2 microA/cm2 (mean +/- SEM, n = 5) and 5.1 +/- 0.3 microA/cm2 (n = 5) in 3-day and 17-day cells, respectively. When the charge carrier was Ba2+, the L-type Ca2+ channel current density was also smaller in 17-day cells (22.7 +/- 1.8 microA/cm2) than in 3-day cells (28.3 +/- 2.1 microA/cm2). In single-channel recordings, the mode-2 behavior was infrequent in 17-day cells compared with 3-day cells. High-open probability sweeps (with an open probability of greater than 0.25), reflecting mode-2 behavior, accounted for 20.2% and 3.7% in 3-day and 17-day cells, respectively. The ensemble-averaged currents in 17-day cells was 37% of that current in 3-day cells. In addition, decay of the averaged current appeared to be faster in 17-day cells than in 3-day cells. All data from the single-channel analysis agreed with the data from the whole-cell voltage clamp.(ABSTRACT TRUNCATED AT 250 WORDS)


ACTH hypertension in the rat: role of sodium chloride. Adrenocorticotrophin (ACTH) produces adrenally dependent increases in both blood pressure and salt (NaCl) appetite in the rat. The present study examines the effect of free access to a high intake of NaCl on ACTH hypertension in the rat to test the hypothesis that high NaCl intake would amplify the rise in blood pressure. Either water or 1%NaCl were offered to sham or ACTH treated Sprague-Dawley rats. Subcutaneous injections of synthetic ACTH (0.5 mg/kg/day) for 10 days caused large increases in the intake of both 1%NaCl (+240 +/- 6 ml/day) and water (+45 +/- 4 ml/day), urine volume (1%NaCl + ACTH +182 +/- 4 ml/day, ACTH + water +36 +/- 2 ml/day), adrenal weight (ACTH + water 176 +/- 18, ACTH + 1%NaCl 367 +/- 129 mg/100 g body weight) and maximum systolic blood pressure (SBP) (ACTH + water +18 +/- 5 mmHg; 1%NaCl + ACTH +16 +/- 3 mmHg). Some ACTH + 1%NaCl rats developed severe edema. Thus, free access to NaCl in ACTH treated Sprague Dawley rats did not potentiate the hypertension, but was associated with edema in some animals.


Studies with guanosine-5'-monophosphate (GMP): new method for measurement and effects on blood pressure. A practical new method for measuring serum guanosine-5'-monophosphate (GMP) was developed and three experiments were performed using this method. In the first, we observed the reduction of blood pressure (BP) and the elevation of serum GMP level persisting for 3 hours in male Japanese White Rabbits administered GMP, 50 mg/kg given as a single oral dose. In the second, 6-week-old male spontaneously hypertensive rats (SHR) received GMP, 200 mg/kg/day, orally for 8 weeks. The systolic BP in the GMP-treated rats, which averaged 170.2 mmHg, was lower than that of the control group, which averaged 188.0 mmHg. Arteriosclerotic findings were milder in the GMP-treated SHR as compared to the control. In the third experiment, the serum GMP level was measured in humans. We observed a significant negative correlation between the serum GMP concentration and systolic or diastolic BP. In conclusion, GMP reduced the BP in experimental animals, suggesting that it may be useful as an antihypertensive agent.


Involvement of the H+/K(+)-ATPase alpha subunit as a major antigenic protein in autoimmune gastritis induced by neonatal thymectomy in mice. Autoimmune gastritis develops spontaneously in approximately 60% of BALB/c mice thymectomized neonatally. Histologically and clinically it is similar to the atrophic gastritis associated with pernicious anaemia in humans. Here we identified antigenic protein relating to the pathogenesis of autoimmune gastritis in these mice. All sera from 32 thymectomized mice with gastritis contained autoantibodies to the vesicular fraction prepared from rat gastric parietal cells. Immunoblot analysis revealed all of these to react with a 94-kD protein corresponding in molecular mass with the H+/K(+)-ATPase alpha subunit. Some sera were also reactive with 65-85-kD and/or 60-kD proteins, whose sizes correspond to the H+/K(+)-ATPase beta subunit and intrinsic factor, respectively. The finding that immuno-adsorption with these sera resulted in reduction of H+/K(+)-ATPase activity in the vesicular fraction, supported a conclusion of H+/K(+)-ATPase alpha and/or beta subunits as the antigenic proteins. After immunization of normal syngeneic mice with various doses of gastric parietal cells or their vesicular fraction, all sera from animals demonstrating atrophic gastric mucosa with lymphocyte infiltration reacted with the H+/K(+)-ATPase alpha subunit. No antibodies to other proteins were induced even in mice immunized with higher doses of antigen. We therefore conclude that H+/K(+)-ATPase alpha subunit is important as the target antigen in pathogenesis of autoimmune gastritis in neonatally thymectomized mice, probably due to a high affinity for the MHC molecule.


The defect seen in the phosphatidylinositol hydrolysis pathway in HIV-infected lymphocytes and lymphoblastoid cells is due to inhibition of the inositol 1,4,5-trisphosphate 1,3,4,5-tetrakisphosphate 5-phosphomonoesterase. Lymphocytes infected in vivo with HIV or lymphoblastoid cells exposed in vitro to either HIV or its envelope glycoprotein (gp120) show a defect in inositol polyphosphate-mediated signal transduction together with an associated abnormality in intracellular calcium regulation. The defect in patients reverses after treatment with the anti-retroviral agent zidovudine (AZT). We present evidence that the defect is at the level of the Ins (1,3,4,5)P4 5-phosphomonoesterase (PME) in these cells and that, though elevation of the intracellular ATP level partially down-regulates the activity of this enzyme, such changes alone are unable to account for the complete inhibition seen in HIV-infected cells.


[Acute relapsing sensory neuropathy following upper respiratory infections--a case report]. A 44-year-old man developed numbness in his hands and feet and an unsteady gait following an upper respiratory symptoms. The symptoms progressed rapidly and reached maximum in a few days, followed by a gradual improvement over a few weeks, but his gait remained unsteady. His symptoms progressively increased over 14 years, with 11 relapses of similar episodes. Neurologic examination showed normal limb strength. Deep tendon reflexes were absent. Touch, cold, and pinprick sensation were impaired in his hands and feet. There was a profound loss of position and vibration sense in all extremities. He demonstrated an unsteady and broad-based gait. Electrophysiologic and histologic examination on the peripheral nerves revealed evidences of axonal degeneration without demyelination involving the sensory nerves.


[Reducing body myopathy--a case report]. A 2-year-old girl with reducing body myopathy was reported. She had no family history of neuromuscular disease. She developed normally with no delay in milestones during infancy. She had no muscle weakness or hypotonia up to 2 years of age when she received mumps vaccination. Three days after the injection, she was first noticed to have limb muscle weakness. The muscle weakness progressed rapidly with increasing difficulty in gait and raising the upper arms, particularly the left. Four months later, she had difficulty in keeping her head up and could no longer climb the stairs. On physical examination, she had proximal dominant generalized muscle weakness, with a preferential neck muscle involvement. She walked waddlingly and stood up with Gowers' maneuver. Facial and ocular muscles were intact. No dysarthria, dysphagia or respiratory difficulty was noted. EMG showed myopathic pattern. Serum creatine kinase level was moderately elevated to 739 IU/l. In the biopsied left biceps muscle, there was marked variation in fiber size, but no apparent necrotic or regenerating fibers. The most striking feature was the presence of numerous eosinophilic inclusions which reduced nitroblue tetrazolium (NBT) and were, therefore, stained dark with menadione-linked alpha-glycerophosphate dehydrogenase even without the substrate of menadione, showing the histochemical characteristics of "reducing" body. The bodies were predominantly seen in fibers with disorganized intermyofibrillar networks and with high acid phosphatase activity. On electron microscopy, the reducing bodies consisted of fine granular material with the similar electron density to the chromatin granules and were located mostly around the degenerated nuclei, suggesting the nuclear degeneration playing a role in forming the reducing bodies.(ABSTRACT TRUNCATED AT 250 WORDS)


[A case of Sotos syndrome associated with peripheral nerve involvements]. A case of the Sotos syndrome associated with peripheral nerve involvements was reported. A 52-year-old male was admitted to Kawasaki Medical School Hospital because of gait disturbance, muscle atrophy, and weakness in both hands. This case was diagnosed as the Sotos syndrome based on the following symptoms and findings, acromegaloid features, hypertrophic changes in the hands and feet, a history of epileptic episodes, a low IQ, a normal growth hormone value, and no tumor lesion in the pituitary gland. Radiological examination disclosed a cauliflower-like appearance of the finger tips and thickness of the heel pads. Brain CT and MRI revealed diffuse mild brain atrophy. An electroencephalogram showed diffuse theta waves with sharp waves in the right parietal region. A needle electromyogram revealed neurogenic change in both upper and lower limbs. A nerve conduction study disclosed the carpal tunnel syndrome and cubital tunnel syndrome. These findings suggest that, as in the case of acromegaly, entrapment neuropathy and peripheral neuropathy can also be induced in the Sotos syndrome.


Neuroendocrine carcinoma of the uterus.

Neuroendocrine carcinoma of the uterus. Neuroendocrine carcinomas of the cervix and endometrium were reviewed. They have been variously designated as carcinoid, argyrophil cell carcinoma, apudoma, small cell carcinoma, oat cell carcinoma, endocrine carcinoma, and neuroendocrine carcinoma, the last-mentioned term being preferred in this chapter. Adenocarcinomas with neuroendocrine cells are occasionally encountered in the cervix and endometrium. It is generally questioned whether they should be included in the spectrum of neuroendocrine carcinomas, although differential diagnosis between some such tumors of the gastrointestinal tract and neuroendocrine carcinoma is reported to be difficult. Since the majority of neuroendocrine carcinomas of the cervix are highly aggressive, it is important to establish the neuroendocrine nature in the cervical carcinomas. In addition to the characteristic histologic features and argyrophil stainability, immunohistochemical demonstration of several neuroendocrine markers may be helpful in diagnosing neuroendocrine carcinoma of the cervix. Ultrastructural demonstration of neurosecretory granules is almost decisive in establishing the tumor's neuroendocrine nature, but it is not applicable in all cases. Neuroendocrine carcinomas of the cervix have been treated by surgery, radiation therapy, and chemotherapy, but optimal treatment methods have not yet been established because of the rarity of the tumor. Finally, we have described a typical neuroendocrine carcinoma of the cervix and reported some data regarding its experimental study.


Dephosphorylation of cdc2 on threonine 161 is required for cdc2 kinase inactivation and normal anaphase. Exit from metaphase of the cell cycle requires inactivation of MPF, a stoichiometric complex between the cdc2 catalytic and the cyclin B regulatory subunits, as well as that of cyclin A-cdc2 kinase. Inactivation of both complexes depends on proteolytic degradation of the cyclin subunit, yet cyclin proteolysis is not sufficient to inactivate the H1 kinase activity of cdc2. Genetic evidence strongly suggests that type 1 phosphatase plays a key role in the metaphase-anaphase transition of the cell cycle. Here we report that inhibition of both type 1 and type 2A phosphatases by okadaic acid allows cyclin degradation to occur, but prevents cdc2 kinase inactivation. Complete inhibition of type 2A phosphatase alone is not sufficient to prevent cdc2 kinase inactivation following cyclin proteolysis. We show further that residue 161 of cdc2 is phosphorylated in active cyclin A or cyclin B complexes at metaphase, whilst unassociated cdc2 is not phosphorylated. Proteolysis of cyclin releases a free cdc2 subunit, which subsequently undergoes dephosphorylation and then migrates more slowly than its Thr161 phosphorylated counterpart in Laemmli gels. Removal of phosphothreonine 161 requires cyclin proteolysis. However, it does not occur even after cyclin proteolysis, when both type 1 and type 2A phosphatases are inhibited. We conclude that both cyclin degradation and dephosphorylation of Thr161 on cdc2, catalysed at least in part by type 1 phosphatase, are required to inactivate either cyclin B- or cyclin A-cdc2 kinases and thus for cells to exit from M phase.


In vitro quantitative autoradiography of [3H]-L-deprenyl and [3H]-PK 11195 binding sites in human epileptic hippocampus. Distribution of the enzyme monoamine oxidase B (MAO-B) and the peripheral benzodiazepine binding site (omega 3 site) was studied by quantitative autoradiography using [3H]L-deprenyl and [3H]PK 11195, two tentative glial markers, as ligands. Sclerotic hippocampus from seven patients who had had anterotemporal lobe resection because of intractable complex partial epilepsy were investigated and compared with postmortem hippocampus from three nonepileptic controls. A significantly higher degree of L-deprenyl and PK 11195 binding was observed in the epileptic cases. The increased binding of both ligands correlated to extent of neuronal loss, but only PK 11195 showed correlation to degree of gliosis. We concluded that both ligands could provide useful markers for quantitating the degree of gliosis in pathologic states such as epilepsy. They may be applicable in future in vivo studies with positron emission tomography (PET).