Role of gelsolin interaction with actin in regulation and creation of actin nuclei in chemotactic peptide activated polymorphonuclear neutrophils. In vitro Ca++ activates gelsolin to sever F-actin and form a gelsolin-actin (GA) complex at the+end of F-actin that is not dissociated by ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) but is separated by EGTA+PIP/PIP2. The gelsolin blocks the+end on the actin filament, but the-end of the filament can still initiate actin polymerization. In thrombin activated platelets, evidence suggests that severing of F-actin by gelsolin increases GA complex, creates one-end actin nucleus and one cryptic+end actin nucleus per cut, and then dissociates to yield free+ends to nucleate rapid actin assembly. We examined the role of F-actin severing in creation and regulation of nuclei and polymerization in polymorphonuclear neutrophils (PMNs). At 2-s intervals after formyl peptide (FMLP) activation of endotoxin free (ETF) PMNs, change in GA complex was correlated with change in+end actin nuclei,-end actin nuclei, and F-actin content. GA complex was quantitated by electrophoretograms of proteins absorbed by antigelsolin from cells lysed in 10 mM EGTA,+end actin nuclei as cytochalasin (CD) sensitive and-end actin nuclei as CD insensitive increases in G-pyrenyl actin polymerization rates induced by the same PMNs, and F-actin content by NBDphallacidin binding to fixed cells. Thirty three percent of gelsolin was in GA complex in basal ETF PMNs; from 2-6 s, GA complexes dissociate (low = 15% at 10 s) and sequentially+end nuclei and F-actin content and then-end nuclei increase to a maximum at 10 s. At > s GA complex increase toward basal and + end nuclei and F-actin content returned toward basal. These kinetic data show gelsolin regulates availability of + end nuclei and actin polymerization in FMLP. However, absence of an initial increase in GA complex or - end nucleating activity shows FMLP activation does not cause gelsolin to sever F- or to bind G-actin to create cryptic + end nuclei in PMNs; the results suggest the + nucleus formation is gelsolin independent.


The relationship between plasma beta-endorphin, opioid receptor activity, and silent myocardial ischemia. To investigate the role of the opioid system in the pathophysiology of silent ischemia through opiate antagonism with naloxone, and to determine the reproducibility of resting and postexercise beta-endorphin levels in predominantly asymptomatic patients with coronary artery disease. Randomized, double-blind, placebo-controlled crossover trial. A University hospital referral center. Ten patients with prior evidence of silent exercise-induced ischemia were studied. An infusion of saline placebo or naloxone at two dose regimens of 0.015 mg/kg or 0.15 mg/kg before supine exercise testing during three separate occasions for each patient. Plasma beta-endorphin was measured at rest, immediately after exercise, and 5 min poststress. Timing and severity of angina and exercise hemodynamics were also determined. Seven of 10 patients reported no angina, whereas the other three experienced angina with placebo and after administration of naloxone at both doses. The severity and duration of angina was consistently noted to decrease in these patients after naloxone administration, especially after low-dose naloxone relative to placebo. There were no apparent correlations between beta-endorphin levels and the characteristics of angina in these three patients, nor between beta-endorphin and hemodynamic responses in all patients in the study. (a) naloxone failed to precipitate angina in this population of patients with silent ischemia; (b) naloxone appears to exert an analgesic effect at low doses; and (c) a variability of 5 pM at rest and 13 pM after exercise might be expected in predominantly asymptomatic patients due to random variation, which is comparable with results found in normal subjects.


Ki-1 positive large cell anaplastic lymphoma: multiple bone lytic lesions and interleukin-6. Ki-1-positive large cell anaplastic lymphoma (Ki-1 LCAL) is recognized as a clinicopathologic syndrome with fever, peripheral lymphadenopathy and cutaneous nodules; the neoplastic cells express Hodgkin's disease-associated antigen, Ki-1 (CD30). We review here a recent case of Ki-1 LCAL with multiple bone lesions with destruction and present additional information. Although bone absorption is reported in some cases of Ki-1 LCAL, the genesis of bone absorption is unclear. Interleukin-6 (IL-6) is an important regulator of osteoclast formation and activation and can induce bone absorption. In our case, the surgically removed tumor tissue was studied for IL-6 mRNA expression and IL-6 secretion without any stimulation. Northern blot analysis showed strong IL-6 mRNA expression in the tumor tissue and ELISA assay showed a large amount of IL-6 in culture supernatants of the tumor tissue. Based on these results, coupled with the reported evidence, we discuss the close relationship between the presence of osteolytic lesions and IL-6 production in Ki-1 LCAL.


Antigenic expression of B-cell chronic lymphocytic leukemic cell lines. A flow cytometric analysis of five B cell chronic lymphocytic leukemic (B-CLL) cell lines was undertaken using 129 unknown reagents from the blind panel (BP) and 72 reagents from the known CD panel obtained from the Fourth International Leucocyte Differentiation Conference and Workshop, B cell section (Vienna, 1989). The five cell lines examined were: SeD (PNAS 75, 5706, 1978), B-CLL-LCL (BLOOD 71, 9, 1988), JVM-HH and JVM-2(INT J CAN 38, 531, 1986), and WR#1 (TH and BD). The reagents were #1-129 (blinded panel) and reagents 1-44 and 53-84 (CD panel with CD23 reagents missing). Positivity was defined as greater than 30% of the cells having a three fold increase or more in mean channel fluorescence. Fourty-three reagents of the blinded panel were negative by these criteria while all remaining reagents were positive on all five lines. SeD showed the lowest reactivity; B-CLL-LCL and JVM-2 showed the most reactivity; JVM-HH and WR#1 were intermediate. The known CD panel confirmed the reactivity of the blinded panel. An average immunophenotype was constructed and compared to published normal EBV lymphoblastoid cell lines and several differences were noted. There was an absence or significant decrease in the expression of CD19, CD21, CD22 and CD37 while there was an increased expression of CD38, CD54, CD74 and CD76. The heterogeneity observed between the B-CLL lines may in part be due to polymorphisms but is more likely to represent the underlying heterogeneity seen in common and familial B-CLL.2+öff


An Italian experience of high-dose chemotherapy and autologous bone marrow transplantation (ABMT) in germ cell tumours. Suggestions for future direction. In this paper an Italian cooperative trial investigates the role of a high-dose regimen with carboplatin, etoposide and ifosfamide in germ cell tumours. Twenty-eight patients underwent one or two transplants. Seventeen with progressive disease. Nine in sensitive relapse and two with stable disease after salvage therapy. Toxicity was generally moderate: two deaths occurred at day 15 from ABMT (one from VOD and one from tumour growth). Five patients are alive and disease free at least 10 months follow-up. In highly pre-treated patients this high-dose combination seems to give an option of cure for relapsed patients. Early transplantation is suggested.


Effects of phorbol esters and cytokines (interleukin-2,-4, and -6) on the proliferation and surface phenotype of Epstein-Barr virus immortalised human B lymphocytes. Epstein-Barr virus (EBV)-induced in vitro infection of peripheral blood mononuclear cells (PBMCs) leads to a polyclonal proliferation and immortalisation of B lymphocytes. In the present study we determined the effects of three different cytokines, interleukin-2 (IL-2), interleukin-4 (IL-4) and interleukin-6 (IL-6), and the tumour promoting phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA) on EBV-immortalised B lymphocytes. These factors have known activities on normal B cells. IL-4 and IL-6 increased significantly EBV-B cell proliferation after 3 and 5 days of culture, where IL-2 had no effect. The effect of IL-4 and IL-6 on EBV-B cells was abolished after pre-incubation with anti-IL-4 and anti-IL-6 neutralising antisera, respectively. TPA induced a dose dependent inhibition of proliferation both in serum free and 10% fetal calf serum (FCS) supplemented culture medium. Combinations of TPA and interleukins did not restore lymphoblastoid cell proliferation to background levels. All possible combinations of the three cytokines showed no synergistic or antagonistic effect on proliferation. TPA induced significant phenotypic changes of EBV immortalised B lymphocytes, by increasing IL-2 receptor (IL-2R) expression and decreasing CD20 and CD23 antigen expression. Other B cell differentiation antigens; HLA-DR, CD19, and transferrin receptor (CD71), did not demonstrate significant changes. A dose dependent inhibition of CD21 and increase in CD22 expression was observed in 2 out of 3 lymphoblastoid cell lines tested.


Coexistence of acute monoblastic leukemia and adult T-cell leukemia: possible association with HTLV-I infection in both cases? A 64 year-old Japanese man who developed acute monoblastic leukemia during the course of adult T-cell leukemia/lymphoma (ATL) was studied. Leukemic cells in the peripheral blood and bone marrow were monoblasts positive for alpha-naphthol butyrate esterase (alpha-NBE) staining, CD11c and CD36 antigens, whereas tumor cells in the pleural effusion were ATL cells positive for CD2, CD4, CD25, CD29 and CD45RA antigens. These two malignant cells had different chromosomal abnormalities. Monoclonal integration of human T-cell leukemia virus type I (HTLV-I) proviral DNA and T-cell receptor C beta gene (TCR C beta) rearrangement were detected in the ATL cells, but not in the leukemic monoblasts. By polymerase chain reaction (PCR) in the peripheral blood mononuclear cells (CD11c+ 98%, CD2+ 4%, CD20+ 0%) not containing ATL cells, the presence of the gag region of HTLV-I was confirmed. These facts indicate that a double positive T cell (CD29+, CD45RA+) was possibly the target cell for HTLV-I infection and that HTLV-I was not directly related to the oncogenesis of the monocyte lineage in the present case, even if it did infect the monocytes. However, there is still an outside possibility that HTLV-I induced acute monoblastic leukemia indirectly.


Human herpesvirus-6 infection and bone marrow transplantation. Human herpesvirus-6 (HHV-6) is ubiquitous in the human population and causes exanthem subitum, a benign disease seen in infancy. However it also produces a wide spectrum of clinical manifestations including cases with a fatal outcome. The virus remains latent in several organs, including the kidneys, liver, lymph/nodes and salivary glands, after the primary infection and reactivates when immune function is impaired. Reactivation of the virus occurred in a half of the bone marrow recipients 2 to 4 weeks after bone marrow transplantation (BMT). It remains to be established whether HHV-6 in fact causes the fever and rash observed in recipients who have reactivation of the virus. The data reviewed here will be required to compare with those of human herpesvirus 7 and a different group of HHV-6.


The beta-carboline abecarnil, a novel agonist at central benzodiazepine receptors, influences saccharin and salt taste preferences in the rat. Abecarnil is a recently described beta-carboline which acts at central benzodiazepine receptors (BZR) and has anxioselective/anticonvulsant properties. While it may be classified provisionally as a partial agonist at BZR, there is also evidence that its pharmacological profile may be due to a selective action at BZR subtypes. The general aim of the present series of experiments was to investigate the effects of abecarnil on ingestional behaviour in the rat. The results indicated that abecarnil (0.3-10 mg/kg, i.p.) selectively increased the intake of preferred 0.05% sodium saccharin and 0.9% sodium chloride solutions in two-choice tests using water-deprived rats. These results confirm and extend previous work with the potent BZR agonist clonazepam. Moreover, abecarnil significantly increased the ingestion of sweetened mash and 3% sucrose solution in nondeprived animals. In general, these results indicate that abecarnil is effective in increasing ingestional responses, a characteristic it shares with classical agonists like diazepam. The results could be accounted for in terms of a partial agonist profile for abecarnil, but do not rule out the possibility of selective actions at BZR subtypes.


Stimulus frequency and intensity: critical determinants of opioid enhancement or inhibition of evoked methionine-enkephalin release. Responses to opioids can be bimodal depending on the concentration of opioid used. For example, low concentrations (nM) enhance whereas higher concentrations (10-100 nM) inhibit the electrically evoked release of enkephalin from the myenteric plexus. The nature of the responsiveness of the enkephalin release process to opioids is also dependent on the intracellular and/or extracellular milieu of enkephalin-containing neurons or other neurons of this plexus. Intracellular levels of cAMP, availability of pertussis toxin- and cholera toxin-sensitive guanine nucleotide binding proteins and intracellular calcium concentration have all been shown to be important determinants of opioid excitatory versus inhibitory actions. The present data indicate that the inhibition of enkephalin release produced by U50,488H or sufentanil is no longer observed when the applied voltage is increased 3- or 2-fold, respectively. Under this condition, a previously inhibitory concentration of opioid produces an enhancement of stimulated enkephalin release. Increasing the frequency of the applied stimulation from 5 to 60 Hz (at a constant voltage) also reverses the sufentanil-induced inhibition to a facilitation of enkephalin release. These data indicate that the intensity (voltage) or frequency of the stimulation that is used to release enkephalin is a critical determinant of the nature of its modulation by opioids. The possible relevance of these findings to known differences in opioid sensitivity between different types of pain is discussed.


Tonic inhibition of striatal dopamine transmission: effects of benzodiazepine and GABAA receptor antagonists on extracellular dopamine levels. At present, it is unclear whether ligands which bind at the benzodiazepine/GABA receptor complex play a tonic modulatory role with regard to striatal dopamine (DA) transmission. The present study was designed to examine the effects of Ro15-1788, a benzodiazepine (BZ) receptor antagonist, and SR 95531, a GABAA receptor antagonist, on striatal extracellular DA (DA[e]) concentrations in anesthetized and awake rats using the technique of in vivo microdialysis. Local administration of Ro15-1788 resulted in a dose-dependent increase in DA[e] in both anesthetized and awake animals. The Ro15-1788-induced increase in DA[e] was blocked by coadministration of the BZ agonist diazepam, as well as GABA. Local administration of SR 95531 also resulted in a dose-dependent alteration in striatal DA levels in both anesthetized and awake animals. The SR 95531-induced increase in DA was blocked by coadministration of GABA. The results suggest that GABA may play a tonic inhibitory role with regard to striatal DA transmission.


Abdominal adipose tissue from broiler chickens selected for body weight or for food efficiency differ in in vitro lipolytic sensitivity to glucagon and to chicken growth hormone, but not to dibutyryl cAMP. 1. Differences in responses to lipolytic agents have been investigated in vitro in abdominal adipose tissue from lines of broiler chickens selected for body weight (GL, a 'fat' line) or for food efficiency (FC, a 'lean' line). 2. Dibutyryl cyclic adenosine monophosphate stimulated in vitro lipolysis, as measured by the glycerol release, by adipose tissue from GL or from FC chickens to the same extent. 3. Glucagon stimulated glycerol release from adipose tissue from FC chickens, but not from GL chickens. 4. Adipose tissue from GL chickens was much more sensitive to chicken growth hormone (GH) compared to FC chickens. 5. It is concluded that the selection criteria applied influenced the number of adipose GH and glucagon receptors, the number of adipose GH receptors being lower and of glucagon receptors being higher in FC chickens compared to GL chickens.


Alterations in carbohydrate metabolism by exogenous dexamethasone and corticosterone in post-hatched White Leghorn chicks. 1. Alterations in carbohydrate metabolism in terms of tissue glycogen contents, phosphorylase (EC 2.4.1.1) activity, hepatic glucose-6-phosphatase (6-6-Pase: EC 3.1.3.9) activity and blood glucose have been evaluated in 30-d-old White Leghorn chicks under induced chronic hypocorticalism (by dexamethasone: DXM) and hypercorticalism (by corticosterone: CORT). 2. DXM treatment showed increased tissue glycogen contents and hypoglycaemia with decreased phosphorylase activity while CORT treatment produced a reverse set of changes. 3. Both steroid treatments increased hepatic G-6-Pase activity. These observations have been taken to indicate a definite role for glucocorticoids in regulating carbohydrate metabolism in neonatal chicks. 4. It is suggested that hypo- or hyper-corticalism could influence carbohydrate metabolism by affecting the secretory/activity ratio of pancreatic hormones.


[Study by in situ hybridization of the prevalence of herpes virus as cofactors of the tumoral development of papillomatous lesions of the anogenital region in seropositive and seronegative men against HIV]. Infection with specific types of HPV has emerged as necessary but not sufficient factor in the neoplastic transformation of anogenital condylomas. Some viruses (HIV, Herpes viridae: HSV, CMV, EBV) might act as cofactors in the neoplastic changes and cancer. To study the prevalence of these viral pathogens in anogenital lesions, biopsies were obtained from HIV seropositive or seronegative men and tested using in situ hybridization technique. Infection by "high risk" HPV, HSV and CMV are facilitated in patients immunocompromised by HIV. Presence of CMV is more frequent in high risk HPV-induced lesions than in low risk HPV lesions.


Protein kinase C mediated activation and phosphorylation of Ca(2+)-pump in cardiac sarcolemma. The effects of purified protein kinase C (PKC) on the Ca(2+)-pumping ATPase of cardiac sarcolemma were investigated. The addition of PKC to sarcolemmal vesicles resulted in a significant increase in ATP-dependent Ca2+ uptake, by increasing the calcium affinity by 2.8-fold (Km 0.14 vs. 0.4 microM for control) and by increasing Vmax from 5 to 6.8 nmol.mg protein-1.min-1. The addition of PKC also stimulated Ca2+ ATPase activity in sarcolemmal preparations. This activity was increased further upon the addition of calmodulin. These results suggest that PKC stimulates Ca2+ ATPase through a kinase-directed phosphorylation. The addition of PKC to a purified preparation of Ca2+ ATPase in the presence of [gamma-32P]ATP resulted in a 100% increase in phosphorylation that was dependent on the presence of Ca2+, phosphatidylserine, and phorbol 12,13-dibutyrate. These results demonstrate that the Ca2+ ATPase of canine cardiac muscle can be phosphorylated by PKC in vitro, resulting in increased affinity of the Ca2+ ATPase for Ca2+ and increase in the Ca2+ pump pumping rate. The results suggest that the Ca(2+)-pumping ATPase in heart tissue can be stimulated by PKC, thereby regulating the intracellular Ca2+ levels in whole heart.


Differential effects of intracellular calcium elevating agents on adrenergic-stimulated cyclic nucleotide and melatonin synthesis in rat pinealocytes. In this study, the role of elevation of intracellular Ca2+ and activation of protein kinase C on adrenergic-stimulated cyclic nucleotide accumulation and melatonin synthesis in rat pinealocytes was investigated. It was found that whereas KCl, ionomycin, and ouabain, three Ca(2+)-elevating agents, had a potentiating effect on adrenergic-stimulated cyclic AMP response, their effects on melatonin synthesis were inhibitory. Similar inhibition was also observed when dibutyryl cyclic AMP was used to stimulate melatonin synthesis. By determining intracellular Ca2+ directly, it was found that the enhancing effects of these agents on the cyclic AMP response but not their inhibitory effects on melatonin synthesis paralleled their abilities to elevate intracellular Ca2+. In comparison, activation of protein kinase C significantly enhanced the adrenergic-stimulated cyclic AMP response and, to a lesser degree, the adrenergic-stimulated N-acetyltransferase and melatonin levels. These results indicate that (i) Ca(2+)-elevating agents have opposite effects on adrenergic-stimulated cyclic AMP and melatonin production; (ii) a post cyclic AMP event of importance to melatonin synthesis is inhibited by these agents; and (iii) the mechanism of inhibition may not be directly related to their effect on intracellular Ca2+.


Effect of lithium chloride on ornithine decarboxylase activity in rat adrenal. The effects of lithium chloride on ornithine decarboxylase (ODC) activity were compared in the adrenal and kidney of control (saline treated) and prolactin-treated rats. ODC activity was decreased in kidney of both groups of animals, the magnitude of the effect of lithium in the hormone-treated group varying with the time of administering the lithium relative to prolactin. The response in the adrenal was quite different. Following treatment with LiCl, there was a gradual increase in ODC activity from a low of 10-35 pmol CO2 x 30 min-1.mg protein-1 in control animals to values 20- to 30-fold greater at 5 h. In rats treated simultaneously with LiCl and prolactin, ODC activity was greater at 5 h than that observed in animals receiving either compound alone, indicating that their effects were additive. When LiCl was given 4 h after prolactin, i.e., 1 h before sacrifice, ODC activity decreased to a very low level at 5 h, as in other tissues. The increase in ODC activity in the adrenal following LiCl is of the same magnitude as the changes observed in tissues stimulated to undergo alterations in proliferation, differentiation, or metabolic or membrane activity by hormones and other external stimuli.


Reduction properties of nitrated naphthalenes: relationship between electrochemical reduction potential and the enzymatic reduction by microsomes or cytosol from rat liver. The nitroreductase activities of rat liver microsomes and cytosol towards various nitrated naphthalenes (1-, 2-mononitro-, 1,3-, 1,5-, 1,7-, 1,8-dinitro-1,3,5- and 1,3,8-trinitronaphthalenes) were characterized as follows. (1) The rates of reduction of nitrated naphthalenes in either microsomal or cytosolic incubation were found to increase in the order of trinitro- > dinitro- > mono-nitronaphthalene, although, in the case of microsomal nitroreduction, trinitronaphthalenes were reduced more rapidly than in cytosol. (2) The effective cofactors, electron donors, in the nitroreduction of nitrated naphthalenes in cytosol were NADH and hypoxanthine, but not NADPH. (3) The nitrated naphthalenes with a nitro group at a beta-position appear to be more easily reduced among the various isomers. The cytosolic nitroreductase activities towards the nitrated naphthalenes were closely related to the single-electron reduction potentials measured by cyclic voltammetry and hence, there was a good relationship between the logarithm of nitroreductase activities and the electrochemical reduction potentials. In microsomes, nitroreductase activities were rather less well related to electrochemical reduction potentials.


Oxidative damage to bovine serum albumin induced by hydroxyl radical generating systems of xanthine oxidase + EDTA-Fe3+ and ascorbate + EDTA-Fe3+. Oxidative damage to bovine serum albumin (BSA) was induced by hydroxyl radical (HO.) generating systems of xanthine oxidase (XO) + EDTA-Fe3+ and ascorbate + EDTA-Fe3+. Formation of bityrosine and loss of tryptophan were observed in the ascorbate + EDTA-Fe3+ system and carbonyl formation was induced by both systems. Mannitol and ethanol very strongly inhibited the carbonyl and/or bityrosine formation, indicating that the oxidative damage to BSA was due to HO(.). The sulfhydryl (SH) groups of BSA were very sensitive to the XO + EDTA-Fe3+ but not to the ascorbate + EDTA-Fe3+ system. Catalase but not hydroxyl radical scavengers or superoxide dismutase strongly inhibited the loss of SH groups, indicating that H2O2 is involved in their oxidation. Fragmentation of BSA was observed during exposure to the XO + EDTA-Fe3+ and ascorbate + EDTA-Fe3+ systems and the products presented a broad band on sodium dodecyl sulfate polyacrylamide gel electrophoresis. Little formation of amine groups was observed in these systems, indicating that little peptide bond cleavage occurred. BSA exposed to the ascorbate + EDTA-Fe3+ system was more readily degraded by trypsin than that exposed to the XO + EDTA-Fe3+ system. Elastase degraded BSA exposed to the ascorbate + EDTA-Fe3+ system but not to the XO + EDTA-Fe3+ system.


Conversion of premalignant human cells to tumorigenic cells by methylmethane sulfonate and methylnitronitrosoguanidine. Nine human tumor cell lines derived from both epithelial and mesenchymal tumors exhibited either an anchorage-independent growth non-tumorigenic phenotype or an anchorage-independent tumorigenic phenotype. Transformed epithelial cell lines with the non-tumorigenic phenotype could be converted to a progressively growing tumor phenotype following treatment with either methylmethane sulfonate (MMS) or N-methyl-N'-nitro-N-nitro-soguanidine (MNNG). In contrast, sarcoma derived cell lines with a non-tumorigenic phenotype could be converted to a progressively growing tumor phenotype only with MNNG. SV40 immortalized HET-1A non-tumorigenic phenotype cells could be converted to a progressively growing tumorigenic phenotype, infrequently, when treated with MNNG, but not MMS. Progressively growing tumors produced by either MMS or MNNG treated non-tumorigenic phenotypes exhibited metastatic potential in nude mice. Chemically treated HET-1A cells acquired the ability to produce tumor in mice but the tumor did not exhibit metastatic potential. In contrast, populations of tumorigenic cells were not rendered more biologically aggressive after treatment with either MMS or MNNG; i.e., the latency period for tumor development was not accelerated and the tumors did not exhibit metastatic potential. These results suggest that the biological effects of MMS and MNNG on non-tumorigenic, tumorigenic and immortalized cell lines are phenotype specific.


In vitro and in vivo toxicity of 2',3'-dideoxycytidine in mice. 3T3 mouse embryo fibroblast cell growth was inhibited in a concentration dependent manner by 2',3'-dideoxycytidine (ddCyd), a strong inhibitor of human immunodeficiency virus. Cell growth inhibition was associated with an increased incorporation of ddCyd into cell DNA. In contrast SP2/0-Ag14 (a mouse myeloma) cell growth is not inhibited by 100 microM ddCyd both in the presence or absence of hypoxanthine and thymidine. Furthermore, in vitro spleen cell proliferation, upon phytohemagglutinin (PHA) addition, was much more affected by ddCyd in C57BL/6 mice than in Swiss albino mice. That indeed ddCyd affects spleen cell proliferation was confirmed by studies on splenocytes obtained from C57BL/6 mice that received ddCyd for 2 weeks in drinking water. These results suggest that ddCyd toxicity in mice is cell and strain dependent and that the toxicity mechanism is related to the incorporation of the drug in cell DNA.


Photochemical DNA modifications induced by 1,2-dioxetanes. 1,2-Dioxetanes are efficient sources of triplet excited carbonyl compounds, into which they decompose on thermal or photochemical activation. In the presence of DNA, the decomposition of dioxetanes gives rise to DNA modifications, which have been studied by means of specific repair endonucleases. Cyclobutane pyrimidine dimers, which are generated by triplet-triplet energy transfer, were detected by a UV endonuclease; they made up between 2% and 30% of the total modifications recognized by a crude repair endonuclease preparation from Micrococcus luteus. For various 1,2-dioxetanes, the yield of pyrimidine dimers was proportional to their triplet excitation flux. DNA strand breaks, sites of base loss (AP sites; recognized by exonuclease III and endonuclease IV) and dihydropyrimidines (recognized by endonuclease III) were found to represent only a small fraction of the modifications. The majority of the modifications detected were recognized by formamidopyrimidine-DNA glycosylase (FPG protein) and represent 8-hydroxyguanine (7,8-dihydro-8-oxoguanine) residues or other yet not defined base modifications which are recognized by this enzyme. The modifications were generated in similar relative yields by thermal and photo-induced decomposition of the 1,2-dioxetanes and therefore emanate under both conditions from the excited carbonyl compounds. The formation of the FPG protein-sensitive modifications was efficiently quenched by azide anions; the Stern-Volmer quenching of these modifications was 150-fold more effective than that of the pyrimidine dimers. The relative amounts of the two types of modifications were strongly dependent on the structure of the 1,2-dioxetanes and on the concentration of molecular oxygen. Singlet oxygen appears to be involved only to some extent in the generation of the FPG protein-sensitive base modifications as their yield was only moderately (approximately 2-fold) increased in D2O as solvent. A mechanism is suggested in which oxidized guanine is predominantly formed by a single-electron-transfer reaction of the triplet excited carbonyl product derived from the 1,2-dioxetane, followed by unknown secondary oxidations, which involve molecular oxygen and/or undecomposed 1,2-dioxetane.


Chemical modification of low-density lipoprotein enhances the number of binding sites for divalent cations. The EPR technique with paramagnetic Mn(II) ions has been used to probe the negatively charged sites on the surface of modified low-density lipoprotein (LDL). LDL modified in five different ways exhibited increased binding capacity for divalent cations. Enhanced binding is caused by the increase in the number of 'strong' binding sites. The 'strong' sites have been identified to be the aspartic acid and/or glutamic acid carboxyl residues and the 'weak' sites are zwitter-ionic phospholipids. In native LDL the negative groups make 'bonds' with the positive lysyl residues, thus stabilizing the structure. Any deprotonation or modification of the lysine amino groups makes the LDL structure more loose and the amino acid carboxyl groups accessible to divalent cations.


Implications of a non-lamellar lipid phase for the tight junction stability. Part II: Reversible modulation of transepithelial resistance in high and low resistance MDCK-cells by basic amino acids, Ca2+, protamine and protons. The transepithelial resistance of confluent epithelial cell monolayers was monitored to investigate the influence of basic amino acids, Ca2+, protamine and protons on tight junction electrical resistance. In an accompanying paper we investigated the effect of these substances on the lamellar/hexagonal II phase transition in reconstituted phospholipid membranes containing phosphatidylserine and phosphatidylethanolamine. We conclude that the permeability of tight junctions may be described by a lipid phase equilibrium where the lamellar phase corresponds to an open state and the hexagonal lipid phase to the closed state of the cell contact. This dynamic lipid model is well suited to describe the morphological as well as functional properties of the tight junctions.


Effects of single and repeated administration of vigabatrin on the performance of rats in a 5-choice serial reaction time task. The present study investigated whether pharmacological stimulation of the GABAergic system can affect attention. The effects of vigabatrin, a novel antiepileptic drug, on the performance of rats in a 5-choice serial reaction time task assessing selective attention were studied. The effects of acute (100, 300, 500 and 1000 mg/kg) and subchronic (50, 100, 200 and 300 mg/kg/day) administration of vigabatrin were investigated. Previous studies have shown that with acute administration of 300-500 mg/kg or subchronic administration of 50-200 mg/kg/day vigabatrin has anticonvulsant activity. At acute doses of 100 and 300 mg/kg or subchronic administration of 50-200 mg/kg/day vigabatrin had no effect on selective attention. At acute dosing of 500 mg/kg, vigabatrin slightly decreased behavioral activity of rats through decreasing the number of trials completed and percent of correct responses. The highest doses used caused an overall behavioral impairment with no marked depletion of any particular function. The results showed that administration of vigabatrin at antiepileptic doses produced no or slight impairment in attentional function. The deficits seen with higher doses were possibly due to a decrease in behavioral activity.


Modifications of the binding properties of the human VIP receptor of IGR39 cells by sulfhydryl reagents. The effects of specific sulfhydryl reagents, N-ethylmaleimide (NEM), p-chloromercuribenzoic acid (PCMB) and 5-5'-dithiobis(2-nitrobenzoic acid) (DTNB), were tested on the vasoactive intestinal peptide (VIP) receptor binding capacity of the human superficial melanoma-derived IGR39 cells. On intact cell monolayers NEM and PCMB inhibit the specific [125I]VIP binding in a time and dose-dependent manner while DTNB has no effect at any concentration tested. Inhibitory effects of NEM and PCMB on high and low affinity VIP receptor are not identical. With NEM-treated cells, only low affinity sites remained accessible to the ligand. Their affinity constant is not modified. With PCMB-treated cells, the binding capacity of high affinity sites is reduced by 56% while the binding capacity of low affinity sites is not significantly affected. For both types of binding sites, the affinity constants remain in the same range of that of untreated cells. On cells made permeable by lysophosphatidylcholine, DTNB is able to inhibit the specific [125I]VIP binding in a time and dose-dependent manner. The three sulfhydryl reagents stabilize the preformed [125I]VIP receptor complex whose dissociation in the presence of native VIP is significantly reduced. Labeling of free SH groups with tritiated NEM after preincubation of cells with DTNB and VIP made possible the characterization of reacting SH groups which probably belong to the receptor. Taken together, these data allow us to define three classes of sulfhydryl groups. In addition, it is shown that high and low affinity sites have different sensibility to sulfhydryl reagents.


The effects of the nitric oxide donors molsidomine and SIN-I on human polymorphonuclear leucocyte function in vitro and ex vivo. The nitrovasodilator and nitric oxide donor molsidomine and its metabolite SIN-I dilate vascular smooth muscle and inhibit platelet activation by increasing intracellular concentrations of cyclic GMP. We have therefore studied the effects of molsidomine and SIN-I on isolated human polymorphonuclear leucocytes (PMN) in vitro and ex vivo. In vitro molsidomine dose-dependently reduced beta-glucuronidase release and the generation of superoxide anions from non-activated and from FMLP- or PAF-stimulated human PMNs. SIN-I was equally effective in reducing beta-glucuronidase release and totally inhibited oxygen radical generation at a concentration of 580 mumol.l-1. In a double-blind, placebo-controlled, randomized trial we also studied beta-glucuronidase release and the generation of superoxide anions from isolated PMNs. Blood was drawn from 12 healthy volunteers before and 3 h after oral molsidomine (16 mg) or placebo. There was no statistically significant difference in beta-glucuronidase release and superoxide anion formation when the PMNs were isolated before or after molsidomine or placebo. This was the case for non-activated, as well as FMLP- or PAF-stimulated PMNs. Thus, the nitric oxide donors molsidomine and its metabolite SIN-I caused a dose-dependent inhibition of PMN functions in vitro, but no significant inhibition when the PMNs were isolated after oral molsidomine.


Differential effect in vitro of tumor necrosis factor-alpha (TNF) on normal and virus-infected erythroid progenitors from Friend virus (FVA)-infected mice. In vivo administration of tumor necrosis factor-alpha (TNF) suppresses both normal and Friend virus (FVA)-infected erythroid progenitor cells (CFU-E). To examine the mechanism of erythroid suppression by TNF, we examined TNF's direct effect on normal and virus-infected cells in vitro. Productively infected fibroblast cell lines, fresh acute virus-infected spleen cells, and virus-infected CFU-E were sensitive, whereas uninfected CFU-E were resistant to TNF cytotoxicity in vitro. When FVA-infected erythroblasts were depleted from the spleen cell population in vitro with antivirus antibodies, TNF suppression of the remaining (uninfected) cells was abrogated. In contrast, both normal and virus-infected macrophage progenitor cells and immature erythroid progenitor cells were equally sensitive to TNF cytotoxicity in vitro. Normal erythroblasts had significantly fewer TNF receptors than FVA-infected erythroblasts, which also were morphologically less mature. These results suggest that TNF can differentially suppress late-stage virus-infected erythroid progenitors in vitro.


Photodamaging effects of merocyanine 540 on neutrophils and HL-60 cells. Merocyanine 540 (MC540) is a photosensitizing dye that has been used in several preclinical models and in a phase I clinical trial for the extracorporeal purging of tumor cells from autologous bone marrow grafts. The mechanism of the cytotoxic activity of MC540 is not yet fully understood, and the subcellular targets of MC540-mediated photodynamic damage remain to be identified. The human neutrophil provides an attractive model with which to study the effects of photoactivated MC540 on several well-defined cellular functions. As we report in this paper, simultaneous exposure of neutrophils to MC540 and light inhibited phagocytosis, random migration, chemotaxis, hydrogen peroxide production, and oxygen consumption. By contrast, the ability of neutrophils to kill engulfed bacteria and to produce superoxide radical was not compromised. Intracellular ATP levels and the activities of the cytosolic enzymes superoxide dismutase, catalase, and myeloperoxidase were only slightly reduced. Even in HL-60 leukemia cells, which bind more dye and are more readily killed by MC540-mediated photodynamic therapy than neutrophils, superoxide dismutase, catalase, and myeloperoxidase activities remained at normal or near-normal levels. These results are compatible with the view that plasma membrane components are primary targets of MC540-mediated photodynamic damage.


Primary somatosensory cortex in rats with pain-related behaviours due to a peripheral mononeuropathy after moderate ligation of one sciatic nerve: neuronal responsivity to somatic stimulation. Single-unit recordings were made under moderate gaseous anaesthesia in the hindpaw representation area of the two primary somatosensory motor cortices (SmI) of rats (n = 58) rendered mononeuropathic by four loose ligatures placed around one common sciatic nerve 2-3 weeks beforehand. The rats exhibited clear hyperalgesia and allodynia from the paw with the ligated sciatic nerve, to both mechanical and thermal stimuli. From the tested neuronal population (n = 640), about the same proportion could be activated by somatic stimuli in each cortex: 165/362 (45%) in the cortex contralateral to the ligated sciatic nerve (Cc), 105/278 (37%) in the cortex ipsilateral to the ligated sciatic nerve (Ci). Neurones driven by light touch, exhibited RFs strictly contralateral to the recording sites. Their proportion and response characteristics were similar regardless of recording side. However, the number of neurones with RFs in the sciatic nerve territory was above 95% in the Ci, and was dramatically reduced to 43% in the Cc. By contrast, the number of neurones with RFs supplied by the saphenous nerve reached 57% on this side. Although the RF size of all the neurones appeared roughly normal, there were fewer Cc than Ci neurones with RFs located on the paw itself and with RFs of extremely small size in the sciatic nerve territory. The proportion of neurones responding to a joint stimulus was significantly higher in the Cc than in the Ci. The neuronal responses to joint stimuli of the paw with the ligated sciatic nerve were significantly more sustained than those recorded in the Ci and elicited from the normal paw. The proportion of neurones driven by mechanical stimulation which gave rise to nociceptive reactions in freely moving animals, i.e. "nociceptive" neurones, was comparable in each cortex. However, half of the Cc neurones exhibited paroxysmal discharges occurring without intentional stimulation and of long duration (1 min to several minutes). Only 66% of Cc but 93% of Ci "nociceptive" neurones were exclusively activated by pinch. The remaining Cc neurones were also activated by applying moderate pressure to the paw with the ligated nerve. Pinch responses from the paw with the ligated nerve were often more intense and of longer duration than responses elicited from the intact paw. The "nociceptive" Cc neurones were especially sensitive to thermal stimuli of 39-44 degrees C when the stimuli were applied to the paw with the ligated nerve. They also responded vigorously to a 10 degrees C stimulus applied to this paw.(ABSTRACT TRUNCATED AT 400 WORDS)


[Prospective comparison between a clinical and biological score and ultrasonography for the diagnosis of benign or malignant hepatic lesions]. The purpose of this study was to compare ultrasound with a clinical and biological score to differentiate benign and malignant liver lesions. The study was carried out prospectively in 100 consecutive patients with liver masses of unknown origin. The accuracy of ultrasound and the score was 84% and 76%, respectively (P = 0.02). Ultrasound and the score gave the same results for the diagnosis of hepatic lesions in 68% of cases. When both ultrasound and the score were in agreement, the prediction of malignancy was 94% ant that of benignity was 100% accurate. Although ultrasound was superior to the score in this series, it is interesting to calculate the score because of the higher predictive values when both methods are associated.


1H-NMR for the safety control of food packaging materials: analysis of extracts from polyolefin samples. A new approach for the identification of the additives in extracts of packaging plastics, using proton nuclear magnetic resonance (1H-NMR), is presented. The technique can be used in a preliminary step for the determination of contaminants potentially released by the food packaging materials; it may greatly reduce the time required to identify the constituents of the materials, either individually or as functional classes. A classification of the EEC additives is proposed on the basis of chemical shift. 1H-NMR can also be used as a fingerprint technique in the quality control of the food packaging materials.


Biological function of DNA methylation.

Biological function of DNA methylation. Structural and functional properties of prokaryotic DNA methyltransferases are summarized. The different aspects of the role of DNA methylation which influences DNA-protein interaction in restriction and modification of DNA and in mismatch repair, DNA replication and gene expression are discussed.


Electron microscopic analysis of two nonconjugative derivatives of plasmid R1drd-19Km- from Escherichia coli. The plasmids pON5300 and pON5304, nonconjugative variants of the plasmid R1drd-19Km, were analyzed by electron microscopy. It was found by heteroduplex mapping that a 1.4 kb DNA segment was inserted into EcoRI E fragment of both plasmids, where some tra-genes and oriT are localized. Although this DNA segment was mapped to the same region its orientation was different in each of the two plasmids. The inserted DNA segment was identified as an IS10R sequence on the basis of analysis of self-annealed molecules of pON5304 and their cleavage with EcoRV restriction enzyme. These methods enable us not only to map IS10R sequences on 87 kb pON5300 and 65 kb pON5304 molecules, respectively, but also to define their orientation.


Analysis of enrofloxacin and its metabolite ciprofloxacin in bovine and porcine muscle by high-performance liquid chromatography following cation exchange clean-up. A simple and rapid method of analysis for the trace residue determination of enrofloxacin and its metabolite ciprofloxacin has been developed. Clean-up of the samples is by cation exchange solid phase extraction (SPE) and determination made by high-performance liquid chromatography using a base-deactivated column and fluorescence detection. The method has been validated for the determination of residues in bovine and porcine muscle tissue and bacon. Recoveries at the 0.010 mg kg-1 level for enrofloaxacin and ciprofloxacin respectively were 90%, 75% in bovine muscle, 75%, 54% in porcine muscle and 81%, 63% in bacon. Determination to the 0.001 mg kg-1 level in bovine muscle and to the 0.002 mg kg-1 level in porcine muscle and bacon was also carried out. The method has been used as a quantitative screening procedure.


Oligounsaturated fatty acid production by selected strains of micromycetes. Fifteen strains of filamentous fungi from the Culture Collection of Fungi (Charles University, Prague) were tested for their lipid production, fatty acid composition with emphasis on accumulation of oligounsaturated fatty acids. All cultures contained palmitic (16:0), palmitoleic (16:1), stearic (18:0), oleic (18:1), linoleic (18:2) and gamma-linolenic (18:3) acid (GLA). The mycelium of Cunninghamella elegans, Rhizopus arrhizus, Mortierella parvispora, M. elongata and M. alpina contained arachidonic acid (ARA) in the range of 2.3-33.5% of the total fatty acids. The strains used in our experiment were capable to accumulate a relatively high amount of intracellular lipid (9.6-20.1% in dry biomass). The highest content of GLA (22.3 mg/g) was found in Mucor circinelloides. The strain of M. alpina containing 47.1 mg/g of ARA could be considered as the best producer of ARA.


Lead in feed incident--multi-element analysis of cattle feed and tissues by inductively coupled plasma-mass spectrometry and co-operative quality assurance scheme for lead analysis of milk. Contaminated cattle feed was imported into the UK in 1989 and resulted in lead toxicity in some animals. Rapid analyses for lead and several other possible contaminating elements were required for feed and cattle tissues. Microwave dissolution of samples with measurement by ICP-MS was used for multi-element determinations. Lead was found to be the major contaminant. Lead levels in milk samples were measured by several laboratories during the crisis and an analytical quality assurance scheme was devised to monitor the quality of the data. The scheme allowed any poorly performing laboratories to be rapidly identified and excluded from the survey.


Evolution of collagen arthritis in mice is arrested by treatment with anti-tumour necrosis factor (TNF) antibody or a recombinant soluble TNF receptor. Immunization of DBA/1 mice with type II collagen within complete Freund's adjuvant leads to arthritis, lasting more than 3 months. Injection of anti-tumour necrosis factor (TNF) IgG, 2 and 3 weeks after immunization prevented the development of arthritis in the following months. This treatment had no effect when started 2 months after induction of the disease. A soluble form of the human recombinant TNF receptor type-beta (rsTNFR-beta), continuously infused at a rate of 20 micrograms/day during the second and third week after immunization, also had a long-term protective effect. Anti-TNF antibody had no effect upon the production of anti-type II collagen antibodies. These results indicate that TNF is critically involved in an early phase of this arthritis.


Production of human monoclonal antibodies to myeloperoxidase. Two mouse-human heterohybridomas secreting human antibodies to myeloperoxidase (MPO) were derived from the peripheral blood of a patient who developed microscopic polyarteritis as the result of long-term treatment with hydralazine. Forty-five immunoglobulin-secreting lines were obtained from the fusion of patient lymphocytes with the CB-F7 heteromyeloma cell line. Of these, two antibodies, one IgG and one IgM, bound to myeloperoxidase in solid phase ELISA and gave a perinuclear staining pattern on ethanol-fixed human neutrophil cytospin preparations. The staining patterns were similar to those seen with serum from the patient. Antigen-inhibition studies revealed that the affinity of the IgG monoclonal antibody was 28 times higher (k = 1.4 x 10(-7)) than the IgM antibody (k = 5 x 10(-5)). Cross-inhibition studies further suggested that the two monoclonal antibodies recognized the same epitope on MPO. Of the other secreting cell lines, none produced antibody which reacted with the panel of autoantigens used for testing. Neither mononuclear antibody reacted with this panel indicating that they were not simply polyreactive natural autoantibodies. These are the first human monoclonal antibodies to native myeloperoxidase to be reported.


Effects of cell differentiation on the synthesis of the third and fourth component of complement (C3, C4) by the human monocytic cell line U937. Association of complement synthesis with cell differentiation in U937 cells was investigated using granulocyte-macrophage colony-stimulating factor (GM-CSF), vitamin D3 and interferon-gamma (IFN-gamma) as differentiation-inducing agents. GM-CSF or vitamin D3 enhanced the synthesis of the third component of complement (C3) by U937 cells, but had no stimulatory effect on the synthesis of the fourth component of complement (C4). IFN-gamma increased both C3 and C4 synthesis by U937 cells. Combination of two of these three agents resulted in synergistic enhancement and all three agents caused maximal enhancement of C3 synthesis. Vitamin D3 enhanced IFN-gamma-induced C4 synthesis by U937 cells. These results were confirmed by ELISA and SDS-PAGE after biosynthetic labelling. GM-CSF, vitamin D3 or IFN-gamma increased the expression of complement receptor type 3 (CR3), one of the markers of monocyte/macrophage differentiation. Two of these agents caused a further increase and all three agents maximal increase in CR3 expression. Since C3 was synthesized in parallel with the degree of CR3 expression, the synthesis of C3, but not C4, by U937 cells is thought to be closely related to cell differentiation. It was reconfirmed that the synthesis of C3 and C4 by U937 cells was independently regulated.


Accumulation of p53 protein correlates with tumour proliferative activity in EBV positive Burkitt's lymphoma. p53 tumour suppressor gene is often found mutated in Burkitt's lymphoma (BL) cell lines and tumours. We analysed 35 BL tumours for the accumulation of p53 protein, and correlated the results with DNA flow cytometric data on the proliferative activity (SPF), and data on the presence or absence of Epstein-Barr virus (EBV) DNA. More than one-third (37 per cent) of the tumours showed accumulation of p53, which was considered to be consistent with mutation of the p53 gene. Tumours that were positive both for EBV DNA and p53 had significantly higher mean SPF than corresponding EBV DNA negative and/or p53 negative tumours. The proportions of tumour cells with accumulation of p53 appeared to correlate with tumour SPF only in EBV DNA positive BLs. However, there was no apparent association between accumulation of p53 and the presence or absence of EBV DNA. These findings are suggestive of multiple pathways in BL tumour progression.


Experience with GM-CSF in the treatment of solid tumors. The efficiency of GM-CSF to reduce myelosuppression after chemotherapy depends on the schedule of administration and the dose of chemotherapy. If conventional chemotherapy doses are given, a seven to ten day administration starting one day after the end of chemotherapy is able to reduce both degree and duration of leucopenia. A later onset is less effective, an earlier one aggravates leuco- and thrombocytopenia. The reduction of myelosuppression is accompanied by a reduction of infection rates and hospitalisation of patients due to these complications. If high-dose chemotherapy is given, GM-CSF does not markedly affect nadir values for leuco- and thrombocytes, but still shortens the duration of leucopenia. This effect is consistently seen after the initial cycles of chemotherapy, but seems to be less pronounced in later cycles. Thus, the growth factor administration allows a treatment intensification mainly by shortening of treatment intervals. Whether these modifications will improve the prognosis of patients with solid tumors is currently being investigated in small cell lung cancer in a German multicenter randomized trial.


Obesity in school children and their parents in southern Iran. Data on school children and their parents in Shiraz, Iran, were used to develop power-type obesity indices, Ip = W/Hp. The optimal value of p was 2.5 for children and 1 for their parents. The smoothed obesity index by age charts are presented using the HRY non-parametric method, which is likely to provide normal ranges of fatness measures for clinical work in Iran. The familial pattern of parent-child obesity was investigated by a preliminary analysis of the distribution of obesity using arbitrarily chosen cut-off points to define obese, normal and lean parents. Then principal component analysis (PCA) was applied to analyse the data as continuous variables, which is found to be an efficient sophisticated technique in clarifying the structure of familial obesity. The analysis also shows no indication of clustering of super-obese families. A structural analysis indicates a direct relationship between obesity of parents and children.


Obesity in Pima Indians. Distribution characteristics and possible thresholds for genetic studies. We examined distribution characteristics of the body mass index (BMI; weight/height; kg/m2) in a sample of 1128 male and 1372 female Pima Indians aged 15-65 years. We found that women had a higher mean and variance of BMI than men. From commingling analyses, we determined that the distribution of BMI could be accounted for either by a single skewed distribution or by a mixture of multiple normal components. These component distributions may be used to define provisional thresholds in selecting families for genetic studies. To ensure genetic segregation of obesity predisposing genes in Pima families will require that some members have BMIs > or = 40 kg/m2.


Obesity stigma reduction in medical students. The purpose of this study was to develop and evaluate an educational intervention designed to modify the stigma held by first-year medical students towards obese patients. The intervention, composed of video, audio and written components, was based on Petty and Cacioppo's elaboration likelihood model. Prior to the course, the medical students held largely accurate beliefs about the causes of obesity, but they still maintained negative stereotypes of the obese as lazy and lacking in self-control. Analysis of students' attitudes toward obese patients five weeks and one year after the course indicates that the intervention was effective. At the five-week assessment, students in the intervention group differed from students in the control group on six of eight measures of attitudes toward the obese. One year after the course, the intervention group was significantly more likely to rate genetic factors as important in obesity and less likely to blame the obese for their condition.


Excess androgenicity only partially explains the relationship between obesity and bone density in premenopausal women. Obese subjects have increased bone density relative to non-obese subjects yet this relationship is not fully understood. We examined whether alterations in sex hormones or binding proteins might explain the effect of obesity on osteoporosis in 83 premenopausal women from the San Antonio Heart Study, a population-based study of diabetes. We measured total testosterone, oestradiol, oestrone, sex hormone binding globulin (SHBG), and serum dehydroepiandrosterone sulphate (DHEA-SO4). Bone density was assessed by a Hologic dual photon absorptometer. Lumbar spine and femoral neck density were positively correlated with body mass index (BMI). In addition, femoral neck density was positively correlated with DHEA-SO4. BMI was negatively correlated with SHBG. After adjustment for sex hormones by multiple linear regression a positive association between bone density and obesity still exists suggesting that the association between obesity and bone density is at least partially independent of sex steroids in premenopausal women.


Subcutaneous adipose tissue blood flow in the abdominal and femoral regions in obese women: effect of fasting. Subcutaneous adipose tissue blood flow (ATBF) was measured by the local clearance of 133Xe from the abdominal and femoral regions of nine individuals with non-endocrine obesity before and after seven days of fasting. Fifteen non-obese individuals served as controls. In the obese group ATBF was similar in the abdominal and femoral regions, 1.7 +/- 0.2 and 1.8 +/- 0.2 ml/min/100 g adipose tissue, respectively. In contrast, in the non-obese group the abdominal ATBF was higher, 4.1 +/- 0.6 and 2.4 +/- 0.2 ml/min/100 g adipose tissue, respectively (P < 0.01). During fasting, ATBF in the abdominal region increased by 45% (P < 0.01), but it remained unchanged in the femoral region. The mechanisms behind the differences in responses to fasting in the two regions are unsettled but may depend on regional differences in lipolytic activity and responses to vasoactive substances. Furthermore, the vasodilator response to fasting in the abdominal region in combination with the higher lipolytic rate in that region may be a pathophysiological factor behind the increased cardiovascular morbidity associated with abdominal obesity.


Relation of abdominal obesity to hyperinsulinemia and high blood pressure in men. The relationships between body fatness, adipose tissue distribution, plasma glucose, insulin levels, lipoprotein levels, and resting blood pressure were studied in 81 men aged 36.0 +/- 3.3 years (mean +/- s.d.) (body mass index (BMI): 27.4 +/- 3.8 kg/m2, percentage body fat: 26.4 +/- 6.6%). Systolic and diastolic blood pressures (BP) were significantly associated with the BMI (r = 0.31, r = 0.33, P < 0.01), the waist circumference (r = 0.33, r = 0.27; P < 0.01) as well as with adipose tissue areas measured by computerized tomography (CT) (0.27 < or = r < or = 0.36, P < 0.01). Furthermore, the relative accumulation of subcutaneous abdominal fat, as estimated by the ratio of abdominal to femoral adipose tissue areas measured by CT, was positively correlated with systolic and diastolic BP (P < 0.01). Fasting plasma insulin level (r = 0.30, P < 0.01) as well as the insulin area measured during an oral glucose tolerance test (0.34 < or = r < or = 0.37, P < 0.01) were significantly correlated with blood pressure. Systolic and diastolic BP were significantly associated with HDL2-cholesterol (C) as well as with the HDL2-C/HDL3-C ratio (-0.24 < or = r < or = -0.34), whereas triglycerides (r = 0.23) and the HDL-C/C ratio (r = -0.23) were significantly correlated with diastolic BP only (P < 0.05). Multivariate analysis indicated that the insulin area was the most important variable associated with blood pressure and that this association was independent of total body fatness and regional adipose tissue distribution. Plasma insulin levels explained 14% and 11% of the variance observed in the systolic and diastolic blood pressures respectively. These results suggest that most of the association between abdominal obesity and high blood pressure is mediated by the hyperinsulinemia and/or the related insulin resistant state.


Association of a DNA polymorphism of the apolipoprotein A-I/C-III/A-IV gene cluster with hypertriglyceridemia in obese people. Hypertriglyceridemia is frequently associated with obesity. In the general Caucasian population, an association of the uncommon S2 allele of a DNA polymorphism of the apolipoprotein (apo) A-I/C-III/A-IV gene cluster with hypertriglyceridemia has been reported. To assess the risk of hypertriglyceridemia associated with the S2 allele in obesity, lipid status and apo A-I/C-III/A-IV genotypes were studied in 90 unrelated Caucasian obese subjects. Age, body mass index, percentage body fat and waist-hip ratio were comparable between genotypes. The frequency of S1/S2 genotype was 35% in the hypertriglyceridemic group versus 11.4% in the normotriglyceridemic group (P < 0.05). The odds ratio of hypertriglyceridemia was 3.7 for obese subjects with the S2 allele and 26.7% of hypertriglyceridemias could be attributed to the S2 allele. Women with the S1/S2 genotype had also significantly higher VLDL- and LDL-cholesterol concentrations. These results suggest that the S2 allele modulates the effects of obesity on lipoproteins and increases the risk of hypertriglyceridemia when obese.


Sexual dimorphism of the autonomic nervous system response to weight loss in obese patients. The sex-related response of the sympathoadrenal system to very low calorie diet (VLCD) with sodium restriction has been studied in a group of 16 obese subjects (8 men; 8 women). Once in sodium balance obese men were different from women in respect to initial body weight, mean daily 4 h urinary excretion of epinephrine (E) and norepinephrine (NE) measured for 48 h. After 20 days of VLCD the weight loss was higher in men than women (P < 0.01), E excretion increased in men more than in women (P < 0.01), and was correlated with weight decrement (r = 0.78; P < 0.01). NE excretion decreased in a similar way in both sexes. The present findings demonstrate sexual dimorphism in catecholamine excretion during VLCD and provide further evidence of the relative autonomy of adrenomedullary secretion from sympathetic nervous system activity.


The relationship between body fat distribution and weight loss in obese adolescent girls. Recent studies have demonstrated an association between the pattern of body fat distribution and the occurrence of cardiovascular risk factors. In this study, we evaluated changes in body fat distribution as defined by several anthropometric criteria during a six week weight reduction programme in 110 obese adolescent girls (mean age 15.2 +/- 0.4 years). The standardized regimen included a mixed diet of 1032 kcal/day and a daily exercise programme of 1-2 h duration. The mean weight loss was 8.6 +/- 2.8 kg, decreasing the body mass index (BMI) from 31.4 +/- 4.7 to 28.2 +/- 4.9 kg/m2 (P < 0.01). The reduction in body weight was accompanied by a significant decrease in the waist-to-hip ratio (WHR) from 0.86 +/- 0.06 to 0.81 +/- 0.05 (P < 0.01). The initial WHR was correlated with the degree of weight loss independent of the initial weight (r = 0.34, P < 0.001). Categorized according to the waist-to-hip ratio girls in the upper tertile (WHR > 0.88) lost significantly more weight than girls in the lower tertile (WHR < 0.80) (9.8 +/- 2.7 vs. 6.8 +/- 2.5 kg, P < 0.01). These findings suggest that girls with an abdominal type of obesity benefit more from a weight reduction programme than girls with a gluteal-femoral type of obesity.


Diurnal rhythms of glycogen metabolism in the liver and skeletal muscle in gold thioglucose induced-obese mice with developing insulin resistance. The circadian rhythm of glycogen metabolism in liver and skeletal muscle was studied in lean and gold thioglucose (GTG) induced-obese mice. The active forms of glycogen synthase (GSI) and phosphorylase (GPa) and the total activity of these enzymes were measured every three hours over a 24 h period in mice fed ad libitum. Hepatic and muscle glycogen content displayed a marked diurnal rhythm that was similar in lean and obese mice. In skeletal muscle the glycogen content, GSI and GPa were not significantly different in lean and obese animals over the 24 h period. The activities of muscle GSI and GPa were constant in both groups despite the diurnal variation in the muscle glycogen content. The absence of an increase in the glycogen content of skeletal muscle despite the pronounced hyperinsulinemia and hyperglycemia in the obese mice, may indicate the degree of insulin resistance in this tissue or the maximal capacity of muscle tissue to store glycogen. In liver, glycogen concentration and total glycogen storage were higher in obese mice. Unlike muscle, both hepatic GSI and GPa underwent significant changes in activity over the 24 h period. Hepatic GSI was lower and GPa was higher in obese mice. The circadian rhythm in enzyme activities was independent of both blood glucose and insulin levels. The total glycogen storage and the activities of total phosphorylase and GPa were significantly increased in the liver from GTG obese mice over a 24 h period and could be implicated in the development of insulin resistance and glucose intolerance in this model of obesity.


Ciglitazone prevents and reverses dexamethasone-induced hyperglycemia in female viable yellow mice. Hypercorticism has been observed in numerous obese and diabetic animal models. Adrenalectomy reduces adiposity, hyperglycemia, hyperinsulinemia, and insulin resistance in these animals. The effects of adrenalectomy can be reversed by glucocorticoid replacement. Male and female viable yellow mice share all phenotypic expressions caused by the viable yellow mutation except that males are hyperglycemic and most females are either normoglycemic or only mildly hyperglycemic. The mechanisms that protect female viable yellow mice from hyperglycemia are not known. Implantation of dexamethasone pellets induced hyperglycemia in female viable yellow mice but had no effect on blood glucose of male viable yellow mice and male and female normal mice. The duration of dexamethasone-induced hyperglycemia correlated to the time endogenous plasma corticosterone levels were suppressed. Plasma insulin levels rose in normal mice but only transiently in viable yellow mice. Ciglitazone prevented and reversed dexamethasone-induced hyperglycemia in female viable yellow mice. Since female viable yellow mice, similar to male viable yellow mice, are obese, hyperinsulinemic and insulin resistant, and since dexamethasone is known to cause insulin resistance, these data suggest that dexamethasone increased insulin resistance to a degree that the protective mechanism was overwhelmed and hyperglycemia was induced. Ciglitazone, a compound known to improve insulin sensitivity, may prevent and reverse dexamethasone-induced hyperglycemia by ameliorating the additional insulin resistance caused by dexamethasone. On a molecular level, since dexamethasone suppresses glucose transport, an insulin-sensitive process in many tissues, whereas ciglitazone and other thiazolidinediones facilitate glucose transport, it is possible that ciglitazone prevents and reverses dexamethasone-induced hyperglycemia by regulating the glucose transport systems in insulin-sensitive tissues.


The bactericidal activity of sparfloxacin. Sparfloxacin was found to display a biphasic response against Escherichia coli, Staphylococcus aureus and Staphylococcus epidermidis in nutrient broth. Its optimum bactericidal concentration was found to be identical for all three species which contrasts with other clinically available fluoroquinolones that are more active against E. coli than against staphylococci. Bacterial protein and RNA synthesis as well as cell division were not found to be essential for all the lethality of sparfloxacin, which hence displays bactericidal mechanism B. However, sparfloxacin was unable to kill bacteria in absence of oxygen.


Effect of fenbufen on the pharmacokinetics of sparfloxacin in rats. The effects of fenbufen on the serum concentrations and penetration into the brain and CSF of sparfloxacin (AT-4140), a new quinolone antibacterial agent, were investigated in rats. At designated times after a bolus iv dose of sparfloxacin 10 mg/kg with or without fenbufen 20 mg/kg, arterial blood, CSF and whole brain were simultaneously collected from each rat. Sparfloxacin concentrations were assayed by HPLC. Serum concentration of sparfloxacin declined bi-exponentially with time and was not changed by coadministered fenbufen. Binding sparfloxacin to serum protein slightly decreased after the coadministration. No elevation of sparfloxacin concentrations was observed in either brain or CSF after coadministration with fenbufen except for only a few time-points. The pharmacokinetic analysis based on the physiological model indicated that fenbufen did not affect the permeability across the blood-brain or blood-CSF barrier. These results suggest that fenbufen may be unlikely to affect the pharmacokinetics, involving the entry into the central nervous system, of sparfloxacin.


A continuous spectrophotometric assay for the activation of plant NAD kinase by calmodulin, calcium(II), and europium(III) ions. A continuous spectrophotometric assay has been developed to quantify the calmodulin, calcium(II) ion, and europium(III) ion dependence of the activation of NAD kinase from pea seedlings. Experimental enzyme activation data are compared with the theoretical curves for the binding of calcium(II) ions to the individual calcium binding sites of calmodulin. These results indicate that the binding of three calcium(II) ions is necessary for activation of plant NAD kinase. Further studies demonstrate that europium(III) ions can replace calcium(II) ions in calmodulin with retention of its ability to activate NAD kinase.


Photochrome-labeling method in study of dynamics of biological systems. The theoretical considerations and experimental evidences discussed in this paper indicate that quantitative study of photochromic processes in labeled objects open up new possibilities for investigating microviscosity and conformation transitions in biological systems. The proposed method of photochrome labeling features higher sensitivity and simplicity, and uses labels which are more stable under physiological conditions compared with traditional spin labels.


Structural analysis of 2',3'-dideoxyinosine, 2',3'-dideoxyadenosine, 2',3'-dideoxyguanosine and 2',3'-dideoxycytidine by 500-MHz 1H-NMR spectroscopy and ab-initio molecular orbital calculations. Solution structure of anti-AIDS drug, 2',3'-dideoxyinosine (ddI) has been assessed by NMR spectroscopy and pseudorotational analysis in conjunction with its analogues: 2',3'-dideoxyadenosine (ddA), 2',3'-dideoxyguanosine (ddG) and 2',3'-dideoxycytidine (ddC). The absence of 3'-hydroxyl groups in these compounds has prompted us to establish the relationship between proton-proton and corresponding endocyclic torsion angles in the 2',3'-dideoxyribofuranose moiety on the basis of five available crystal structures of 2',3'-dideoxynucleosides. A subsequent pseudorotational analysis on ddI (1), ddA (2), ddG (3) and ddC (4) shows that the twist C2'exo-C3'-endo forms of sugar are overwhelmingly preferred (75-80%) over the C2'-endo envelope forms. The phase angles (P) for North and South conformers with the corresponding puckering amplitude (psi m) for ddI (1), ddA (2) and ddG (3) are as follows: PN = 0.1 degrees, PS = 161 degrees and psi m = 34.1 degrees for ddI (1); PN = 1.4 degrees, PS = 160 degrees and psi m = 34.2 degrees for ddA (2) and PN = 2.4 degrees, PS = 163 degrees and psi m = 33.6 degrees for ddG (3). The predominant North conformer of ddC (4) is intermediate between twist C2'-exo-C3'-endo and C3'-endo envelope (P = 10.9 degrees) with a psi m of 34.7 degrees. Note that these preponderant North-sugar structures (approx. 75-80%) found in the solution studies of ddI (1), ddA (2), dG (3) and ddC (4) are not reflected in the X-ray crystal structures of 2',3'-dideoxyadenosine and 2',3'-dideoxycytidine. The constituent sugar residues in both of these crystal structures denosine and 2',3'-dideoxycytidine. The constituent sugar residues in both of these crystal structures are found to be in the South-type geometry (ddA crystalizes in C3'-exo envelope form, while ddC adopts the form intermediate between the C3'-exo envelope and C3'-endo-C4'-exo twist form). This means that X-ray structures of ddA (2) and ddC (4) only represent the minor conformer of the overall pseudorotamer population in solution. An assumption that the structure of the pentofuranose sugar (i.e. P and psi m) participating in conformational equilibrium described by the two-state model remains unchanged at different temperatures has been experimentally validated by assessing five unknown pseudorotational parameters with eight unique observables (3J1'2', 3J1'2", 3J2'3', 3J2'3", 3J2"3', 3J2"3", 3J3'4' and 3J3"4') for 2',3'-dideoxynucleosides.(ABSTRACT TRUNCATED AT 400 WORDS)


Brachial plexus injury caused by impalement. Open injuries of the brachial plexus are rare. One such case, that of a 68-year-old impaled on a fence spike, is presented here. Certain principles to guide evaluation and treatment are discussed. Concomitant injury to the pleura or to vascular structures requires immediate attention; the extent and type of plexus damage may be determined from physical findings and the nature of injury. The results of plexus reconstruction are variable and routine exploration may be detrimental. The Brooks classification is reviewed.


Inhibition of salvage synthesis of nucleic acid by adenosine 3',5'-cyclic decylphosphoramidate in mastocytoma P-815 cells. Constant exposure of mastocytoma P-815 cells to adenosine 3',5'-cyclic decylphosphoramidate (1), which is permeable to the cell membrane and resistant to the action of phosphodiesterase, caused a dose-dependent (1 to 50 microM) inhibition in the synthesis of DNA and cell proliferation. Pretreating the cells with compound 1 (20 microM, 4 h) caused considerable inhibition of the incorporation of [3H]thymidine ([3H]TdR) into [3H]deoxythymidine 5'-triphosphate ([3H]dTTP) and that of [14C]hypoxanthine into nucleic acid, but not the synthesis of [14C]dTTP from [U-14C]aspartate. These results indicate that compound 1 preferentially inhibits the salvage synthesis of intracellular nucleotides and nucleic acids. Thymidine kinase, a key enzyme in salvage synthesis of nucleotides, was almost undetectable in cells pretreated with compound 1 at 20 microM for 4 h or at 5 microM for 15 h. On the other hand, compound 1 activated partially purified cAMP-dependent protein kinase A from bovine heart. Judging from these observations, it is likely that compound 1 readily permeates the cell membrane, activates cAMP-dependent protein kinase, then inhibits the salvage synthesis of nucleotides and nucleic acids by inhibiting thymidine kinase, which results in the inhibition of cell growth.


Evidence that potassium channels mediate the effects of serotonin on the ocular circadian pacemaker of Aplysia. The eye of the marine mollusk Aplysia californica contains a photo-entrainable circadian pacemaker that drives an overt circadian rhythm of spontaneous compound action potentials in the optic nerve. Serotonin is known to influence the phase of this ocular rhythm. The aim of the present study was to evaluate whether potassium channels are involved in effects on the ocular circadian rhythm. Our experimental approach was to study the effect of the potassium channel antagonist barium on serotonin-induced phase shifts of this rhythm. The application of barium was found to block serotonin-induced phase shifts whereas barium alone did not cause significant phase shifts. The effects of barium were found to be dose dependent. In addition, barium blocked forskolin-induced phase advances but did not interfere with serotonin-induced increases in cAMP content. Finally, barium antagonized serotonin-induced suppression of compound action potential activity. These results are consistent with a model in which the application of serotonin phase shifts the ocular pacemaker by causing a membrane hyperpolarization which is mediated by a cAMP-dependent potassium conductance.


Color Doppler imaging of inferior vena cava: identification of tumor thrombus. Color-coded Doppler sonography (CCDS) was used for the examination of tumor thrombus (n = 7) and benign thrombosis (n = 6) of the inferior vena cava. Tumor thrombus was due to hepatocellular carcinoma in two cases and to renal cell carcinoma in five cases. Whereas no specific information about the nature of thrombus formation could be gained by gray scale sonography, a typical patchy vascularization pattern was noted within tumor thrombi in six of seven cases using CCDS. This ws due to marked neovascularization within the tumor thrombi, confirmed by histologic examination in all cases. In one patient no vascularization within the tumor thrombus could be observed by CCDS because the thrombus was relatively small. In patients with bland thrombosis, either no vascularization of the thrombus was seen (n = 5) or linear recanalization of the thrombus occurred (n = 1). Therefore, it might be possible to differentiate tumor from nontumor thrombus in the inferior vena cava by using CCDS. Further studies are needed to confirm this preliminary hypothesis.


Genital human papillomavirus infection in women. To enhance nurse clinicians' knowledge of genital human papillomavirus infection in women. Several literature searches using the following terms, dating back to 1986: human papillomavirus (HPV), females, human, cervical neoplasia, risk factors, condylomata acuminata, detection, epidemiology, pathology, psychology, Papanicolaou test, immunosuppression, HIV infection, and AIDS. Forty-three formal research studies regarding the association of various types of HPV infection with cervical intraepithelial lesions, the putative precursor lesions for cervical neoplasia; the outcomes of diagnostic techniques for HPV types; the outcomes of diagnostic/screening techniques for abnormal cervical cells; the association of risk factors for acquiring HPV infection; or the outcomes of therapy. Some additional references were chosen for their presentation of epidemiologic or surveillance data, others for their scientific discussions on related topics. Data were abstracted according to summary measures of the parameter of interest in the sample studied. In most instances, it was the prevalence of HPV, cervical neoplasia, or frequency of use of screening tests. Immunosuppressed clients are at particular risk for HPV-mediated cervical neoplasia. Because Papanicolaou tests are an effective screening tool, cervical cancer is easily detectable. The nurse may facilitate treatment. This is an especially important issue for young women, among whom sexual activity is growing--with attendant increases of HPV and HIV infection.


Inhibition of inositol-1,4,5-trisphosphate-5-phosphatase by chlorpromazine and related compounds. The effects of chlorpromazine and related compounds upon soluble and particulate Ins(1,4,5)P3-5-phosphatase were investigated in rat GH3 pituitary and in human IMR-32 neuroblastoma cells. Chlorpromazine inhibited both soluble and particulate activities with an IC50 of 1 mM after a preincubation time of 10 min at 37 degrees C. The inhibition was time-dependent and could not be reversed by washing the membranes. Inhibition of soluble activity was found at mM concentrations with a number of phenothiazine derivatives with an apparent requirement for a-Cl, an-SCH3 or a-CF3 (compared with an-H or a-COCH3) substituent at the ring position 2. Such a requirement was also seen in other tricyclic compounds, although clozapine was not active and methixene was active. No such requirement was seen for the particulate enzyme.


Gastrointestinal effects of modified guar gum and soy polysaccharide as part of an enteral formula diet. Dietary fiber supplementation may improve gastrointestinal tolerance and decrease diarrhea in patients receiving enteral formula diets. To compare the effects of two dietary fibers on bowel function parameters and short-chain fatty acid excretion we fed 11 healthy men three defined enteral formula diets in random order for 18 days each. The test diets consisted of a fiber-free formula and daily intakes of maltodextrin (0 g of fiber), 15 g of total dietary fiber as an enzymatically modified guar gum, and 15 g of total dietary fiber as soy polysaccharide. Data were also collected while subjects consumed self-selected diets for 5 days. Mean transit time was longer and fecal moisture content was lower on 0 g of fiber and modified guar than on the self-selected and soy diets. Furthermore, mean transit time was slightly longer and fecal nitrogen excretion greater on modified guar compared with 0 g of fiber. Daily fecal output and frequency of defecation were greater, fecal pH was lower, and fecal butyrate concentrations were higher on the self-selected diet compared with the enteral formula diets. However, there was no difference in these parameters among the three liquid diets. Thus, despite significant differences in mean transit time, few differences in other parameters of bowel function were observed when healthy subjects consumed enteral formula diets containing 0 g of fiber and 15 g of total dietary fiber as modified guar and soy.


Pattern of ovarian tumours among Malaysian women at General Hospital, Kuala Lumpur. The objective of this two year retrospective study is to find out the pattern of ovarian tumours among Malaysian women. A total of 280 cases were reviewed. Of these 193 were benign, 81 were malignant and six cases belonged to borderline malignancy. In the general population, equal distribution of serous and mucinous tumours among the benign (15.4%) and malignant (4.3%) types is quite a striking feature. The teratomas were the commonest benign tumour among the Malays and Chinese. Serous cystadenomas were the commonest among the Indians. The Malays had higher incidence of malignant epithelial tumours whereas the Chinese had a higher incidence of metastatic and germ cell tumours. Endometroid tumours occurred from an earlier age of thirty years. There was a preponderance of mucinous tumours among the borderline variety.


Plastid DNA sequence homologies in the tobacco nuclear genome. The tobacco (Nicotiana tabacum) nuclear genome contains long tracts of DNA (i.e. in excess of 18 kb) with high sequence homology to the tobacco plastid genome. Five lambda clones containing these nuclear DNA sequences encompass more than one-third of the tobacco plastid genome. The absolute size of these five integrants is unknown but potentially includes uninterrupted sequences that are as large as the plastid genome itself. An additional sequence was cloned consisting of both nuclear and plastid-derived DNA sequences. The nuclear component of the clone is part of a family of repeats, which are present in about 400 locations in the nuclear genome. The homologous sequences present in chromosomal DNA were very similar to those of the corresponding sequences in the plastid genome. However significant sequence divergence, including base substitutions, insertions and deletions of up to 41 bp, was observed between these nuclear sequences and the plastid genome. Associated with the larger deletions were sequence motifs suggesting that processes such as DNA replication slippage and excision of hairpin loops may have been involved in deletion formation.


Studies on the Mx transposable element system in maize recovered from X-irradiated stocks. The unstable mutant bz-x3m arose in a plant subjected to X-irradiation. The element at the bronze locus is non-autonomous and recombination data indicate that an autonomous element is tightly linked. The autonomous element has been designated Mx (mobile element induced by X-rays) and the non-autonomous element, rMx (responder to Mx). Linkage data indicate that a second Mx lies near the end of the short arm of chromosome 9; in one plant, an Mx that is unlinked was detected. Distinguishing characteristics of bz-x3m are a large window of time in endosperm development during which somatic reversions can arise and a wide range in the frequency at which they occur; these features are heritable. With increasing doses of bz-x3m and Mx, the window expands and the frequency range increases. In kernels containing the bz-x3m allele and the tightly linked Mx, breakage occurs in chromosome 9 distal to the C locus, resulting in breakage-fusion-bridge patterns for endosperm markers that lie proximal to the break. The frequency of breaks and the developmental time at which they occur exhibit the same dosage effect as the somatic reversions of the bz-x3m allele. These observations suggest that an rMx (designated rMxBr) that causes chromosome breakage is positioned distal to the C locus. At the molecular level, the bz-x3m allele is associated with a 0.5 kb increase in fragment size in DNA samples digested with BglII, EcoRI, HindIII and PstI; in germinal revertants, the fragment size returns to that of the progenitor.


[Segmental reflexes after sciatic nerve damage in rats given thyroxine]. Amplitude of the L5 segmental reflex responses 20 min. before and after intraperitoneal introduction of obsidan, clopheline and finoptine was recorded in experiments on spinalized and thyroxinized rats which 3 weeks before have endured transection of the sciatic nerve. It is demonstrated that obsidan does not change the amplitude of the monosynaptic reflex responses of the ventral root restored under the effect of thyroxine at the operated side, while clopheline and finoptine mainly reduce it.


[Management of malnourished children. Cases from the Protestant Hospital of Dabou]. Malnutrition due to lack of nutritional resources unfortunately remains an endemic which ravages Third World countries. Not only is malnutrition a result of continuous droughts and wars, but it is a reflection of the difficult economic circumstances which persist in these countries. Malnutrition should be specifically treated in each region with the least costly locally available means. The experience of the Protestant Hospital of Dabou proves that this manner of care is possible in the midst of a rural area with very limited resources. Hospital surveillance, nutritional education and a stay in the nutritional rehabilitation center offers these malnourished children a new chance to begin in life again. Unfortunately for some, the acquired benefits disappear once they have left the establishment and the child returns to the same least favorable socio-economic environment for his development.


[Electroencephalogram changes and psychopathologic disorders in 60 cases of chronic palm brandy intoxication (Ivory Coast)]. The aim of the study was to assess the effects of chronic palm alcohol (a compound of ethyl and methyl alcohols distilled from palm tree wine) consumption, on waking electroencephalogram (EEG) of 60 alcoholics of both sexes. Four profiles have been determined on EEG abnormalities criteria: 1/ theta rhythms (44 patients), 2/ slow alpha rhythms (9 patients), 3/ slow waves and 4/ paroxysmal rhythms. The whole results are globally in agreement with reports regarding waking EEG abnormalities in ethylic subjects. Nevertheless the presence of thêta waves in recordings of many of the subjects, all irritable or aggressive and exhibiting emotional disturbances, would constitute the sign of this alcoholic type.


Demonstration of a bimodal coaggregation reaction between Porphyromonas gingivalis and Treponema denticola. This study demonstrates a strong coaggregation reaction between 2 suspected periodontopathogens: Porphyromonas gingivalis and Treponema denticola. Other black-pigmented oral bacterial species tested did not coaggregate with T. denticola. This specific interbacterial aggregation was bimodal, since heating of both cell types was required to completely eliminate the reaction. The coaggregation reaction occurred between pH 4 and 9. Under some conditions, arginine and D-galactosamine were effective in preventing the coaggregation. The heat-sensitive receptor on P. gingivalis was found to be loosely bound and could be released by a light ultrasonic treatment of the cells. It is suggested that the bacterial interaction described may participate in the establishment of a potentially pathogenic subgingival plaque.


[Sequela of hand burns in children: 32 cases followed and treated in the Plastic Surgery and Reconstruction department of the University Hospital Center of Treichville, Abidjan]. An investigation on the effects of hand burns has been undertaken about 32 children admitted in the plastic surgery outpatients Department. Accident occurs among the lower class population and heat is the major factor pointed out. The most frequent effects are hand palmar contracture. The surgical treatment completed with the functional re-education has given very good results in 79% cases. However the cheloid contractures are the most difficult to cure. Indeed the authors insist or the prevention and the creation of a center of burned patients and a plastic surgery Service to take care of patients whose life prognostic is optimistic.


Molecular forms of alpha-melanocyte-stimulating hormone in the canine pituitary anterior and intermediate lobe. Reverse-phase high pressure liquid chromatography (HPLC) and radioimmunoassay (RIA) were used to determine the distribution of naturally occurring forms of alpha-melanocyte-stimulating hormone (alpha-MSH) in acid extracts of pars intermedia (PI) and anterior lobe (AL) tissue from canine and rat pituitary. Similarly, intracellular and secreted forms of alpha-MSH were determined using cultured canine PI and AL cells. Rat PI tissue contained predominantly diacetyl-alpha-MSH, while monoacetyl-alpha-MSH was the most abundant form in canine PI. In both canine and rat AL tissue extracts desacetyl-alpha-MSH was the major form of alpha-MSH. The profile of alpha-MSH contained in and secreted into culture medium by canine PI cells was found to be very similar to that in PI tissue extracts. The proportion of monoacetyl-alpha-MSH and diacetyl-alpha-MSH secreted by cultured canine AL cells and contained in extracts of AL cells in culture, however, was much higher than that in tissue extracts. These results indicate that in the dog, as in all other mammalian species studied, acetylated forms of alpha-MSH predominate in PI tissue, while nonacetylated alpha-MSH is the major form in AL tissue. It appears, however, that acetylation of alpha-MSH may occur in cultured canine AL cells, possibly as a result of the absence of factors that normally inhibit acetyltransferase in vivo or as a consequence of culture conditions.


Reflex gastric motor inhibition caused by intraperitoneal bradykinin: antagonism by Hoe 140, a bradykinin antagonist. Bradykinin (BK) has been reported to have mixed excitatory/inhibitory effects on gastrointestinal motility. The present study examined the mechanism responsible for the inhibition of gastric motor activity caused by intraperitoneal administration of BK. Gastric motor activity was measured by recording the intragastric pressure (IGP) of phenobarbital-anesthetized rats via a transesophageal catheter. To facilitate the study of inhibitory influences, gastric motility was stimulated by neurokinin A (NKA), which on intravenous injection evoked reproducible gastric contractions as measured by a rise of IGP. Intraperitoneal injection of BK (0.1-10 nmol) inhibited the NKA-induced increase in IGP in a dose-dependent manner, and the effect of epigastric administration of BK was not significantly different from that of intraperitoneal administration. The inhibitory effect of intraperitoneal BK on gastric motility was due to an effect on BK2 receptors because it was blocked by prior intraperitoneal injection of the BK2 antagonist Hoe 140. The specificity of this BK antagonist was demonstrated by its inability to antagonize the effect of intraperitoneal hydrochloric acid (HCl), which, like BK, inhibited the NKA-induced gastric contractions. Because the BK- and HCl-induced inhibition of the NKA-induced rise of IGP was abolished by acute removal of the celiac-superior mesenteric ganglion complex, but left unaltered by acute bilateral subdiaphragmatic vagotomy, it is inferred that intraperitoneal BK inhibits gastric motor activity via activation of an autonomic reflex that involves prevertebral ganglia.


Stimulation by melanocortins of neurite outgrowth from spinal and sensory neurons in vitro. Using ELISAs for B-50/GAP43 and neurofilament (NF), we tested ACTH(1-24), alpha-MSH, ACTH(4-10), and an ACTH(4-9) analogue (ORG2766) for their ability to induce sprouting and neuritogenesis from spinal and sensory neurons. Dissociated fetal rat spinal cord neurons or neonatal rat dorsal root ganglion (DRG) cells were cultured with peptide and assayed after 24, 48, or 96 h. In spinal neurons, alpha-MSH and ACTH(1-24) induced the expression of B-50 dose dependently. After 24 h alpha-MSH had a stimulatory effect (from 10 nM onwards), with a maximum at 100 microM (36% increase). After 96 h the maximal effect of 100 microM alpha-MSH on B-50/GAP43 was lower (19%). ACTH(1-24) (100 microM) stimulated B-50/GAP43 by 19%. Neurofilament levels (96 h) were elevated maximally by 64% at 100 microM alpha-MSH. In DRG neurons a bell-shaped dose-response curve was found for alpha-MSH, the maximal effect being observed after 48 h at 100 nM: 54% for B-50/GAP43 and 22% for NF. In both culture systems neither ACTH(4-10) nor ORG2766 was effective. We conclude that alpha-MSH stimulates the expression of B-50/GAP43 (sprouting) and the formation of NF (neurite elongation) and may therefore be considered a neurotrophic factor.


Differential effect of arachidonic acid on the vasoactive intestinal peptide receptor/effector system in rat prostatic epithelium during sexual maturation. The effects of alterations in the membrane lipid environment on vasoactive intestinal peptide (VIP) binding and VIP-stimulated cyclic AMP accumulation have been analyzed by arachidonic acid treatment of prostatic epithelial cells from rats at puberty and maturity, two critical developmental periods with characteristic lipidic and androgenic statuses. Treating cells with 0.1 mM arachidonic acid for 15 min at 37 degrees C increased the affinity of VIP receptors and the potency of the neuropeptide (up to five times) in the formation of cyclic AMP at maturity, but not at puberty. The average plasma membrane fluidity (as measured by fluorescence polarization of diphenylhexatriene) remained unmodified after arachidonic acid treatment of cells. The modifications observed in mature rats were specific for the VIP receptor/effector system, since cyclic AMP stimulation by isoproterenol or forskolin was not affected by cell treatment with arachidonic acid. These results are compatible with the existence of a particular lipidic microdomain surrounding the VIP receptor in the cell membrane that would be altered by exposure to arachidonic acid (either directly or through conversion of arachidonic acid to its metabolites, as suggested by experiments on inhibition of the arachidonic acid cascade). This would make it possible for the activation of protein kinase C to phosphorylate VIP receptors in cells from mature rats, but not in those from pubertal animals with a very different membrane lipid composition (as suggested by the corresponding values of membrane fluidity and transition temperature).


Probing the opioid receptor complex with (+)-trans-SUPERFIT. II. Evidence that mu ligands are noncompetitive inhibitors of the delta cx opioid peptide binding site. Previous studies delineated two classes of delta binding sites; a delta binding site not associated with the opioid receptor complex, termed the delta ncx site, and a delta site associated with the opioid receptor complex, termed the delta cx site. The delta ncx site has high affinity for [D-Pen2,D-Pen5]enkephalin, and is synonymous with what is now identified as the delta 1 binding site. Pretreatment of membranes with the delta-selective acylating agents FIT, or (+)-trans-SUPERFIT, deplete membranes of the delta ncx binding site, which permits the selective labeling of the delta cx binding site with [3H][D-Ala2,Leu5]enkephalin. The present study compared the properties of the delta cx binding site present in brain membranes pretreated with (+)-trans-SUPERFIT with the properties of the delta cx site present in untreated membranes. The major findings are: 1) pretreatment of membranes with (+)-trans-SUPERFIT decreased the IC50 values of delta-preferring drugs, and increased the IC50 values of mu-preferring drugs, for the delta cx binding site; 2) the degree of delta selectivity was highly correlated with the magnitude of the (+)-trans-SUPERFIT-induced shift in the IC50 values; 3) the ligand-selectivity patterns of the mu and delta cx sites present in (+)-trans-SUPERFIT-pretreated membranes were poorly correlated; 4) whereas mu-preferring drugs were noncompetitive inhibitors of [3H][D-Ala2,Leu5]enkephalin binding to the delta cx site, delta-preferring drugs were competitive inhibitors.(ABSTRACT TRUNCATED AT 250 WORDS)


Biliary tract invasion and obstruction by hepatocellular carcinoma: report of five cases. Major biliary tract obstruction caused by tumour invasion is a rare manifestation of hepatocellular carcinoma. The authors had the opportunity to diagnose and treat five such cases, three of whom had features of acute cholangitis. The prevalence of both hepatocellular carcinoma and recurrent pyogenic cholangitis is high in patients from the Far East. The former may first present under the guise of the latter. Gastroenterologists and surgeons should be aware of hepatocellular carcinoma when managing these patients who present with obstructive jaundice, gross hepatomegaly and cholangitis.