Inhibition of rat peroxisomal palmitoyl-CoA ligase by xenobiotic carboxylic acids.

ATP-dependent coenzyme A (CoA) ligases catalyse the formation of the acyl-CoA thioesters of xenobiotic carboxylic acids and the formation of xenobiotic-CoAs has been implicated as being a causative factor in peroxisomal proliferation. In this study we have demonstrated using rat liver peroxisomes that the formation of palmitoyl-CoA is inhibited by a variety of xenobiotic carboxylic acids. Palmitoyl-CoA formation exhibited biphasic kinetics indicative of two isoforms, a high affinity (Km1 2.3 microM) low capacity form and a low affinity (Km2 831 microM) high capacity form. These forms were differentially inhibited by a range of xenobiotics. However, it would appear that the low affinity component may not contribute to any major extent to the formation of xenobiotic-CoAs in vivo. At a concentration of 1 mM, greater than 20% inhibition of the high affinity form was observed with the 2-arylpropionates, ibuprofen, naproxen, benoxaprofen, fenoprofen, indoprofen, ketoprofen, tiaprofenic acid and cicloprofen, the hypolipidaemics, nafenopin and ciprofibrate, and the herbicides, silvex and 2,4,5-trichlorophenoxyacetate. Valproic acid, clofibric acid, salicylic acid and 2,4-dichlorophenoxy-acetate were non-inhibitory at all concentrations studied (0.1-2.5 mM). Analysis of the type of inhibition established that only nafenopin (Ki 430 microM) and ciprofibrate (Ki 97 microM) were competitive inhibitors of palmitoyl-CoA formation suggesting that they bind at the active site and thus potentially function as alternative substrates for the peroxisomal ligase. Notably, clofibric acid which has previously been shown to form clofibroyl-CoA in peroxisomes did not interact with the palmitoyl-CoA ligase thereby suggesting that activation is mediated via an alternative peroxisomal CoA ligase. In addition, the xenobiotic inhibitors of the peroxisomal palmitoyl-CoA ligase differed from those previously reported for the equivalent microsomal enzyme suggesting that the organellar forms may be functionally distinct. This study establishes that numerous xenobiotic carboxylic acids interact with the peroxisomal palmitoyl-CoA ligase; however, it would appear that relatively few function as alternative substrates. The toxicological ramifications of peroxisomally mediated xenobiotic-CoA formation and the identification of other peroxisomal xenobiotic-CoA ligase(s) remain to be elucidated.


Studies on the structure of the ligand-binding site of the brain D1 dopamine receptor.

A series of group-specific modifying reagents were tested for their effects on [3H]SCH23390 binding to brain D1 dopamine receptors in order to identify amino acid residues at the ligand binding site of the D1 dopamine receptor that are critical for ligand binding. The dependence of ligand binding on the pH of the incubation medium was also examined. The histidine-selective reagent, diethylpyrocarbonate did affect ligand binding but this is probably not due to an effect at the ligand binding site. Experiments with N-acetylimidazole and ethylacetimidate indicated that modification of tyrosine and amino residues did not exert major influences at the ligand binding site. The use of the thiol dithiothreitol indicated that breakage of a disulphide bond altered ligand binding, probably by affecting the receptor conformation, and the use of the sulphydryl reagent 5,5'-dithio-bis-nitrobenzoic acid showed that modification of a sulphydryl group on the receptor inhibited ligand binding. The carboxyl reagent N,N'-dicyclohexyl carbodiimide (DCCD) potently inhibited ligand binding and the effect could be prevented by occupancy of the receptor site by an agonist or antagonist so that there is an important carboxyl group at the receptor binding site. The total number of D1 receptors was reduced after the modification by DCCD and 70% of the residual receptors showed a reduced affinity for binding [3H]SCH23390, the remainder having the same affinity as untreated receptors. [3H]SCH23390 binding is also reduced by a decrease of pH and this effect seems to depend on the protonation of a group of pKa 6.9. Saturation analysis of [3H]SCH23390 binding performed at pH 7.5 shows a single class of high affinity sites whereas at pH 6.0, two classes of sites with higher and lower affinities are seen. These studies suggested a model whereby [3H]SCH23390 binding is to two receptor isoforms with different pH dependencies for [3H]SCH23390 binding.


Soybean trypsin inhibitor and beta-amylase induce alveolar macrophages to release nitrogen oxides.

Rat alveolar macrophages incubated with soybean trypsin inhibitor and beta-amylase produced nitrite in a dose- and time-dependent manner. This production depends on the presence of L-arginine (L-arg) in the culture medium. The precursor of this nitrite was demonstrated as being nitric oxide by bleaching ferredoxin at 410 nm when added to the culture medium. NG-Monomethyl-L-arginine and the tetrahydrobiopterin biosynthesis inhibitor 2,4-diamino-6-hydroxypyrimidine inhibited the release of nitrite in a dose-dependent manner. Dexamethasone was able to modulate this release. These data indicate that alveolar macrophages are capable of secreting L-arg-derived nitrogen oxides when stimulated with certain alimentary proteins.


[Ginkgo biloba extract in peripheral arterial diseases. Meta-analysis of controlled clinical studies].

In the first part the statistical methods of meta-analysis are discussed. Meta-analysis is considered as a statistical tool for quantitatively summarizing the results of clinical trials with comparable aims (treatments) and designs. Meta-analysis can be based on the significance probabilities or effect values. The last procedure is preferable as it gives an estimate (and confidence interval) for the global effect of the treatment of interest, if homogeneity of the effects between the trials can be assumed. Such a homogeneity can be often achieved by a suitable standardization of the effect variables within the trials. In the second part the methods of meta-analysis are applied to controlled clinical trials with Ginkgo biloba extract EGb 761 in patients with peripheral arterial disease. Included were 5 placebo-controlled clinical trials with similar design and inclusion criteria. In all studies treatment effect was quantified by the increase of walking distance (measured in standardized treadmill exercise). The effect value of EGb 761 treatment was expressed by the standardized mean difference in walking distance increase between EGb 761 and placebo, standardized by the standard deviation. It could be shown that this effect value is homogeneous in all trials. The global effect size was estimated as 0.75. This means that the mean increase in walking distance achieved by EGb 761 is 0.75 times of the standard deviation higher than that achieved by placebo. This value is highly significant different from zero. So the meta-analysis revealed a highly significant therapeutic effect of EGb 761 for the treatment of peripheral arterial disease.


Phosphodiesterase inhibition by enoximone in preparations from nonfailing and failing human hearts.

The effects of enoximone (MDL 17043, Perfan, CAS 77671-31-9) on the activities of the phosphodiesterase (PDE) isoenzymes I-IV and on force of contraction were investigated in ventricular preparations isolated from failing (end-stage myocardial failure, NYHA IV) and non-failing human hearts. In both tissues four PDE isoenzymes (PDE I-IV) with similar properties were separated by DEAE-sepharose chromatography. The effects of enoximone on PDE I-IV activities did not differ between non-failing and failing human hearts. As compared to PDE I (IC50 2100 mumol/l) and II (IC50 2900 mumol/l) enoximone is a selective PDE III (cGMP-inhibited PDE, IC50 5.9 mumol/l) and PDE IV (cGMP-insensitive PDE, IC50 21.1 mumol/l) inhibitor. Milrinone, 3-isobutyl-1-methylxanthine (IBMX) and UD-CG 212 Cl, a derivative of pimobendan, were studied in the failing heart for comparison. Milrinone inhibited PDE I-IV activities similar to enoximone, revealing IC50 values for inhibition of PDE III and IV (1.2 and 3.3 mumol/l) which were about two orders of magnitude lower than that of PDE I and II (173 and 306 mumol/l). UD-CG 212 Cl was the most potent (IC50 0.05 mumol/l) and most selective PDE III inhibitor tested (IC50 for PDE I, II and IV were 175, 181 and 40.8 mumol/l, resp.), whereas IBMX inhibited PDE I-IV nonselectively (IC50 15.3, 26.2, 5.6, 5.8 mumol/l, respectively). In trabeculae carneae from nonfailing and failing human hearts enoximone increased force of contraction only marginally by 18.0 +/- 9.1% (n = 8) and 24.5 +/- 8.7% (n = 9) of the predrug value.(ABSTRACT TRUNCATED AT 250 WORDS)


A molecular model of MHC class-I-restricted antigen processing.

Cells of higher vertebrates have evolved mechanisms that allow a sample of their intracellular contents to be available for surveillance by the immune system. This display of intracellular material is in the form of peptides bound to cell surface major histocompatibility complex (MHC) class I molecules. In this review, John Monaco presents a model of the mechanisms by which this takes place, based on the recent identification of a number of new genes in the MHC.


[The T-lymphocyte antigen receptor].

The precise knowledge of the T-cells antigen receptor (TCR) is of paramount importance; it is the first structure involved in the antigen (allergen) recognition, provided this one is presented in the right conditions; that is in the context of HLA molecules present at the surface of macrophages, after being processed inside. The TCR alpha/beta, present on more than 90% of peripheral T cells, is formed of two glyco-protein chains of similar molecular weight. The cytoplasmic end of the TCR is two short to transmit the message of recognition. The signal is transduced by neighbouring molecules forming the CD3 complex. Other membrane proteins such as CD2, LFA1, reinforce adhesion between immuno-competent cells. The presence or absence of CD4 or CD8 surface antigens, permit to distinguish two T cell subpopulations, namely helper and suppressor/cytotoxic lymphocytes. The TCR gene organization is very similar to that of light and heavy chains of immunoglobulins. Their fortuitous rearrangement explains the very large diversity of the T-cell repertoire. The TCR gamma/delta, although first appeared on the thymic cells, is present on less than 5% of peripheral lymphocytes, where its exact role is still unknown.


[Cytokines and allergy].

The immune system is a homeostatic system adaptating the internal self to the variations of the external non-self and allowing us to get on well together with the environment. Intercellular communications are necessary for the system well operating and are mediated by either direct contacts or by soluble factors called cytokines. Allergy is the upshot of a disregulation of the immune response that does not discriminate between a potentially harmful antigen (tetanus toxoid) and a rather harmless antigen (pigeon droppings). This could be the result of an imbalance between two helper T lymphocytes subsets called TH1 and TH2: their main produced cytokines, respectively IFN-gamma and IL-4, have opposite effects on IgE synthesis whereas other cytokines (IL-3, IL-5) are more directly involved in the differentiation of eosinophil and mast cells lineages.


Observations, legends, and conjectures concerning restricted T-cell receptor usage and autoimmune disease.

It has become clear over the past few years that a variety of experimental autoimmune conditions are mediated by T cells bearing a highly restricted subset of antigen receptors. This restricted TcR usage raises important questions concerning not only the recognition of autoantigens, but also the pathogenic mechanisms underlying many models of autoimmunity. Furthermore, the extension of these findings in certain cases to human disease has raised the possibility of specific therapeutic immune intervention. In this review, we examine the available data on restricted T-cell receptor usage in autoimmune disorders and explore the interpretations and the theoretical and practical implications of these findings.


Muscle pathology in idiopathic cricopharyngeal dysphagia. Enzyme histochemical and electron microscopic findings.

The structural changes in the cricopharyngeal muscle (CM) were examined ultrastructurally and by enzyme histochemistry in five patients suffering from idiopathic cricopharyngeal dysphagia (ICD). Diagnosis was established by fiberoptic esophagoscopy, esophageal manometry and cineradiography. Cricopharyngeal myotomy was performed with marked improvement in all patients. Intraoperatively, a biopsy was taken from the CM. Additionally, all patients underwent neurological examination for possible generalized muscle disease, and a biopsy was taken from a limb muscle. CM from nine cadavers without known history of dysphagia served as control. The control samples disclosed structural changes which were considered to be pathological in other skeletal muscles, and required that the criteria for CM pathology we modified accordingly. In three patients changes in CM histology suggested specific pathogenesis: one patient had evidence for a generalized myositis but was only symptomatic for dysphagia. Another patient had muscle fiber atrophy and slight inflammation in her CM, possibly due to alcohol abuse. The third patient had loss of CM fibers with replacement by connective tissue enough to cause functional disturbances. In two patients no cause for dysphagia was found in either immunohistochemistry or electron microscopic studies. These results demonstrate the special structural features of the CM and indicate that ICD can have multiple etiologies.


Uptake of sufentanil, alfentanil and morphine in the lungs of patients about to undergo coronary artery surgery.

We have studied the pulmonary extraction and retention of sufentanil, alfentanil and morphine using a double indicator technique in 30 patients undergoing elective aortocoronary bypass surgery. Patients were allocated to three groups (10 each) to receive sufentanil 43 micrograms, alfentanil 672 micrograms or morphine 1887 micrograms, mixed with indocyanine green as indicator. After sufentanil, mean peak extraction was 93.7% (95% confidence interval 87.1-100.4%) and release occurred after 16.1 (13.8-18.4) s; first-pass retention was 61.1 (51.3-70.9)%. After alfentanil, peak extraction was 67.4 (42.7-92.0)% and release occurred after 9.9 (8.5-11.3) s; first-pass retention was 10.1 (3.5-16.7)%. After morphine, peak extraction was 58.3 (44.4-72.2)% and release occurred after 7.2 (4.4-10.1) s; first-pass retention was 7.1 (-4.7-19.9)%. Both peak extraction and first-pass retention were significantly greater after sufentanil. There was no significant difference in the peak extraction and first-pass retention between alfentanil and morphine.


Efficacy of orally administered ondansetron in the prevention of postoperative nausea and vomiting: a dose ranging study.

In a placebo-controlled, double-blind study, we have compared the efficacy of ondansetron 16 mg, 8 mg and 1 mg administered 8-hourly for prevention of postoperative nausea and vomiting. We studied 995 patients undergoing major gynaecological surgery; 982 were included in the analysis. Study medication was administered 1 h before induction of anaesthesia and second and third doses were given 8 and 16 h after the first. The treatment groups were similar for patient characteristics, surgical procedures, anaesthetics administered and opioids given. The frequency of nausea was 75%, 70%, 56% and 55% after placebo and ondansetron 1 mg, 8 mg and 16 mg, respectively; the corresponding frequencies of vomiting were 60%, 55%, 37% and 37%. Ondansetron 8 mg was as effective as 16 mg and both resulted in significant reductions in nausea and vomiting compared with placebo and ondansetron 1 mg (P less than 0.001).


Abnormal CD45R expression in patients with common variable immunodeficiency and X-linked agammaglobulinaemia.

This study has investigated the patterns of membrane 2H4 (CD45RA) and UCHL1 (CD45RO) expression by CD4+ and CD8+ lymphocyte subpopulations in 10 adults and seven children with common variable immunodeficiency (CVI) and X-linked hypogammaglobulinaemia (XLA). Of the 10 adults (CVI, n = 8; XLA, n = 2), only one with a diagnosis of CVI showed normal CD45R expression. For the remaining seven adult CVI patients the abnormal CD45R profiles were primarily associated with CD4+ lymphocytes in three and CD8+ lymphocytes in four. Specific increases in CD45RA- CD45RO+ fractions were found in four CVI patients and in all of these there was a concomitant reduction in circulating CD19+ B-cell numbers. Two of the CVI cases were identical twin sisters and both had the same CD45R abnormality. The two adults with XLA also showed abnormal CD45R expression (increased CD45RA+ CD45RO- components) but of particular note, in view of the fact that these were brothers, one was associated with the CD4+ subpopulation and the other with the CD8+ fraction. Similar analyses for the paediatric group revealed, in distinct contrast to the adult patients, no significant abnormalities of CD45R expression suggesting that these defects may not become apparent until a later age. Further investigations of in vitro lymphocyte proliferation (PHA and PWM) in the eight adults with CVI showed a significant correlation between abnormal responses and disordered CD45R expression. It is proposed that these abnormal patterns of CD45R expression by lymphocyte subpopulations in CVI and XLA may be of fundamental importance with respect to the pathogenesis of these particular immunodeficiencies.


Heparin-associated thrombocytopenia in a patient treated with polysulphated chondroitin sulphate: evidence for immunological crossreactivity between heparin and polysulphated glycosaminoglycan.

Heparin-associated thrombocytopenia (HAT) type II, a severe side effect of heparin therapy, is thought to be induced by an immunological mechanism. By crossreactivity studies we have demonstrated that sera of patients with HAT type II activate platelets in vitro not only after the addition of heparin but also after addition of a chemically polysulphated chondroitin-like substance, Arteparon, used for treatment of degenerative joint disease. In addition here, we describe a patient who developed deep venous thrombosis and pulmonary embolism following administration of Arteparon and typical HAT type II with thrombocytopenia, 36 h after the first administration of heparin. This patient had never received heparin, but had repeatedly been treated with Arteparon for degenerative joint disease. We conclude that this patient had been presensitized by Arteparon, as indicated by his clinical course. In vitro studies again confirm crossreactivity between heparin and Arteparon.


Characterization of stable beryllium fluoride, aluminum fluoride, and vanadate containing myosin subfragment 1-nucleotide complexes.

Beryllium and aluminum fluorides are good phosphate analogues. These compounds, like orthovanadate, form stable complexes with myosin subfragment 1 (S1) in the presence of MgADP. The formation of the stable S1-nucleotide complexes is characterized by the loss of ATPase activity. For the complete loss of ATPase activity there was necessary a higher concentration of aluminum than of beryllium or vanadate. In the presence of MgATP the onset of the inhibition is delayed, which indicates that stable complexes cannot form when a specific site is occupied by the gamma-phosphate of ATP or by P(i) derived from the gamma-phosphate. The half-lives of the S1-MgADP-(BeF3-), S1-MgADP-(AlF4-), and S1-MgADP-Vi complexes at 0 degrees C are 7, 2, and 4 days, respectively. In the presence of actin the rate of decomposition of all of the complexes is significantly enhanced; however, the order of decomposition is reversed, the fastest rate being observed with beryllium and the slowest with aluminum. The formation of the S1-MgADP-(BeF3-) and S1-MgADP-(AlF4-) complexes is accompanied by an increase in tryptophan fluorescence similar to that observed upon addition of MgATP to S1. The fluorescence increase develops rather slowly, by suggesting that the rate-limiting step in the formation of the stable complex is an isomerization. The rate of the fluorescence change accompanying the formation of the Be complex is faster than that for the Al complex. Addition of vanadate to S1 causes a static quenching of the tryptophan fluorescence.(ABSTRACT TRUNCATED AT 250 WORDS)


Chloride transport of yeast vacuolar membrane vesicles: a study of in vitro vacuolar acidification.

Effects of various solutes on acidification inside the vacuolar membrane vesicles of the yeast Saccharomyces cerevisiae were examined. ATP-dependent acidification was stimulated by the presence of chloride salts. There was essentially no difference in the stimulatory effects of NaCl, KCl, LiCl, and choline chloride. The membrane potential across the vacuolar membrane was reduced by the presence of Cl- salts. Transport of 36Cl- is driven by the protonmotive force across the vacuolar membrane. Kinetic analyses have revealed that the stimulatory effect of Cl- on internal acidification depends on two distinct components. One shows linear dependency on chloride concentration and is inhibited by 4,4'-diisothiocyano-2,2'-stilbenedisulphonic acid (DIDS). The other exhibits saturable kinetics with an apparent Km for chloride of 15-20 mM. We conclude that the vacuolar membrane of yeast is equipped with Cl- transport systems contributing to the formation of a chemical gradient of protons across the vacuolar membrane by shunting the membrane potential generated by proton translocation.


Inhibition of mitochondrial F1-ATPase activity by binding of (2-azido-) ADP to a slowly exchangeable non-catalytic nucleotide binding site.

F1-ATPase was treated so that it contained three tightly bound nucleotides per molecule. One of these was bound at a catalytic site and was rapidly exchangeable, the two remaining nucleotides were nonexchangeable. Incubation of this preparation with ADP in the presence of Mg2+ results in 40-45% inhibition of the ATPase activity. With 2-azido-ADP instead of ADP, the ligand was covalently bound to F1 by illumination, in the presence or absence of turnover of the enzyme, and the site of binding was determined. In this way, one site could be identified, which induces the inhibition. The attachment of the covalently bound 2-nitreno-ADP is at Tyr-368 of a beta-subunit, characterized in the literature as a non-catalytic site. A second, non-catalytic site also binds 2-azido-ADP, but this binding is partially reversed by the addition of ATP and does not cause further inhibition of the ATPase activity. It is concluded that the slowly exchangeable non-catalytic site is the site of inhibition by ADP.


Toxigenic type A Pasteurella multocida as a causative agent of nasal turbinate atrophy in swine.

Although no clinical signs of atrophic rhinitis (AR) were recognized in 2- and 5-week-old pigs, approximately 60% of 2- to 6-month-old pigs showed clinical signs of AR in an affected pig farm. None of the pigs had normal turbinate at slaughter. Bordetella bronchiseptica was not isolated from any of the pigs before onset and incipient stage of the outbreak (2-week to 2-month-old). Pasteurella multocida of capsular type D was not isolated from any of those pigs. However, toxigenic P. multocida of capsular type A was isolated from a number of the pigs immediately before onset and incipient stage of the outbreak. Thirty-six-day-old primary specific-pathogen-free pigs were inoculated intranasally with a toxigenic type A P. multocida isolated from a 5-week-old pig. Severe nasal turbinate atrophy was observed in those pigs which were necropsied at 3 weeks post-inoculation. This is the first report on outbreak of severe nasal turbinate atrophy induced by toxigenic type A P. multocida in Japan.


Non-cholinergic mechanisms underlying the acute lethal effects of P = S type organophosphorus insecticides in rats.

Intravenous administration of the lethal dose of diazinon or fenthion, P = S type organophosphates, to urethan anesthetized rats induced bradycardia and transient apnea followed by a decline of blood pressure, and death. We investigated the mechanisms of the lethal action of these organophosphates in rats through measurements of blood pressure, heart rate, and respiratory pattern. We compared their cardiorespiratory effects in the five different conditions under anesthesia; 1) normal (without treatment), 2) artificially ventilated, 3) vagotomized, 4) atropinized, 5) pithed, vagotomized and atropinized. It was found that the administration of 200 mg/kg of fenthion and 100 mg/kg of diazinon, caused sudden bradycardia, transient apnea and gradual decline of blood pressure in the anesthetized normal rat, and the rat died. The rats in other conditions also died except the artificially ventilated rats, in which 400 mg/kg of fenthion was administered to cause hypotension and subsequent death. Hypotension was observed consistently even after the cardiac effect such as bradycardia was eliminated by atropine treatment. In the pithed rats which were further vagotomized and atropinized, these organophosphates also caused hypotension. These results may indicate that hypotension is the main cause of death which resulted from intravenous administration of the P = S types. Hypotension may be caused by peripheral cardiovascular effect of the P = S types, which is unrelated to cholinergic mechanisms.


Low order correlations of lipoprotein(a) with other blood lipids and with coagulation and fibrinolysis parameters in hypertensive and diabetic patients.

Lipoprotein(a) (Lp(a)) has been established as an important independent risk factor for the development of cardiovascular disease. Apolipoprotein(a), together with apo B-100 the apolipoprotein of Lp(a), is homologeous to plasminogen but lacks fibrinolytic capacity and appeared to interfere with fibrinolysis in in vitro and ex vivo experiments. We determined the correlations between Lp(a) and other blood lipids (serum cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides), coagulation parameters (fibrinogen, factor VII, factor VIII:C fibrin monomers, thrombin-antithrombin III) and fibrinolysis parameters (tissue plasminogen activator antigen, plasminogen activator inhibitor-1 and D-dimer) in 54 patients with essential hypertension, in 65 non-insulin-dependent diabetic patients and in 116 insulin-regulated diabetic patients. Signs of activated coagulation and increased reactive fibrinolysis were found in all three patient groups. In the hypertensive patients, Lp(a) was significantly correlated with LDL-cholesterol (r = 0.25, P = 0.04) and triglycerides (r = -0.30, P = 0.03), while in insulin-regulated diabetics, Lp(a) was also correlated with LDL-cholesterol (r = 0.20, P = 0.03). In the hypertensive patients and both diabetic groups there was no correlation of Lp(a) with coagulation or fibrinolysis parameters. These data show that Lp(a) concentrations are not related to coagulation or fibrinolysis parameters in hypertensive or diabetic patients and confirm the presence of an activated coagulation system in these patient groups.


Effect of calmodulin on sarcoplasmic reticulum Ca(2+)-ATPase isolated from cardiac muscle.

A Ca(2+)-dependent ATPase, purified from cardiac microsomal membranes by solubilization and chromatography, is identified as cardiac sarcoplasmic reticulum ATPase on the basis of its electrophoretic mobility and its trypsin digestion pattern. The ATPase (both in membranous and purified form) is stimulated by calmodulin, while the skeletal muscle ATPase is not. Rapid kinetic experiments demonstrate that the calmodulin stimulation is already present within the first enzyme cycle following the addition of ATP, and consists of an increased turnover of the phosphorylated enzyme intermediate. The calmodulin effect does not involve the phosphorylation of any protein other than the ATPase. Following the incubation of ATPase with [gamma-32P]ATP, even in conditions of calmodulin stimulation, radioactive phosphorus is found only on the ATPase electrophoretic band, corresponding to the phosphorylated enzyme intermediate. These observations, together with the results obtained for [125I]calmodulin binding to the ATPase, suggest that the stimulation in turnover produced by calmodulin on the ATPase is due to a direct effect on the enzyme. This may provide an independent regulation of the cardiac sarcoplasmic reticulum Ca(2+)-ATPase, in addition to the known regulation mediated by other accessory proteins.


Nucleotide sequence of cDNA clones encoding the beta subunit of mitochondrial ATP synthase from the green alga Chlamydomonas reinhardtii: the precursor protein encoded by the cDNA contains both an N-terminal presequence and a C-terminal extension.

cDNA and genomic clones encoding the beta subunit of mitochondrial ATP synthase from Chlamydomonas reinhardtii have been isolated using heterologous DNA probes from the photosynthetic bacterium Rhodospirillum rubrum. The protein encoded by the cDNA is 79-83% identical to corresponding proteins from higher-plant and mammalian mitochondria, and 75% identical to the R. rubrum protein. It contains both an N-terminal presequence and a unique C-terminal extension. The presequence, which is the first mitochondrial presequence determined in C. reinhardtii, is similar in structure to mitochondrial presequences from other organisms. As chloroplast presequences from C. reinhardtii also share features with mitochondrial presequences from other organisms (L.-G. Franzén et al., FEBS Lett 260 (1990) 165-168), this raises interesting questions about protein targeting to chloroplasts and mitochondria in C. reinhardtii. The possibility that the C-terminal extension is involved in targeting the protein to the mitochondrion is discussed. Southern blot analysis indicates that the protein is encoded by a single-copy gene.


Developmental and environmental concurrent expression of sunflower dry-seed-stored low-molecular-weight heat-shock protein and Lea mRNAs.

We have cloned and sequenced three different cDNAs from sunflower seed-stored mRNA. Sequence similarities and response to heat-shock identified one of the cDNAs as a low-molecular-weight heat-shock protein (lmw-HSP). The other two clones showed significant sequence similarity to the cotton and carrot late-embryogenesis-abundant (Lea) proteins D-113 and Emb-1, respectively. The three cDNAs showed similar expression patterns during zygotic embryo development, as well as in vegetative tissues of 3-day-old seedlings in response to stress. Maximal accumulation of all three mRNAs was detected in dry seeds and during embryo mid-maturation stage, in the absence of exogenous stress. In seedlings, mRNAs accumulated to lower levels in response to osmotic stress and exogenous abscisic acid (ABA) treatments. A differential time course of response to osmotic stress was observed: lmw-HSP mRNA accumulation was induced earlier than that of Lea mRNAs. The coordinate accumulation of Lea and lmw-HSP transcripts during embryo development and in response to stress and ABA suggests the existence of common regulatory elements for Lea and lmw-HSP genes, and supports the notion that HSPs might have alternative functions in the plant cell.


[Diagnosis of chronic backache in health resort patients].

Conservative treatment of backache requires an accurate diagnosis because sometimes such patients are suffering from a serious but unrecognised disease of the abdomen or low back region. Diagnoses like cervical syndrome, lumbar syndrome or even cervicodorso-lumbar syndrome are not sufficiently conclusive to start conservative treatment, since they convey nothing in respect of cause, prognosis or aim of treatment. Patients undergoing a course of inpatient curative treatment of, say, four weeks in a hospital, need special therapy programmes that are meaningfully based on precise data on their disorder. It is important to set great store by the physical examination (which is often performed cursorily before the patients are referred to curative inpatients treatment), including a neurological examination and in some cases also EMG, special radiographs, myelography, CT, magnetic resonance imaging and the like to gather more information on the real cause of the low back pain. This will also help the physician in arriving at a fair estimation of the patient's performance rating. A useful classification of patients into two groups would be as follows: a) without neurological symptoms, and b) with signs of a nerve-root compression or even spinal cord symptoms.


A complete sequence of the rice sucrose synthase-1 (RSs1) gene.

Using a fragment of the maize sucrose synthase gene Sh-1 as probe, the rice genome was shown to contain at least three genes encoding sucrose synthase. One of these genes was isolated from a genomic library, and its full sequence, including 1.7 kb of 5' flanking sequence and 0.9 kb of 3' flanking sequence, is reported. The new rice gene, designated RSs1, is highly homologous to maize Sh-1 (approx. 94% identity in derived amino acid sequence), and contains an identical intron-exon structure (16 exons and 15 introns). Both RSs1 and maize Sh-1 show similar sequence homologies to a second rice sucrose synthase gene described recently (designated RSs2, Yu et al. (1992) Plant Mol Biol 18: 139-142), although both the rice genes predict an extra 6 amino acids at the C-terminus of the protein when compared to the maize gene. The RSs1 5' flanking sequence contains a number of promoter-like sequences, including putative protein-binding regions similar to maize zein genes.


Serine/threonine protein kinases.

Signal transduction in the nervous system is heavily dependent on the three multifunctional serine/threonine protein kinases, PKA, PKC, and CaM-KII. Recent studies have furthered our understanding of how the multiple isoforms of these kinases and their subcellular localizations, regulatory properties, and substrate determinants are important for the specificity of kinase functions.


The involvement of the intestinal microflora in the expansion of CD4+ T cells with a naive phenotype in the periphery.

It is well known that immune reactivity declines with age. Recently, we demonstrated that the age-related decrease in IL-2 production by CD4+ T cells was accompanied by an increased production of IL-4 and interferon-gamma (IFN-gamma). This age-related shift in the profile of lymphokine production was related to phenotypic changes within the CD4+ T-cell subset, that is, a decrease in the percentage of CD45RB++ CD4+ T cells and an increase in the percentage of Pgp-1+ CD4+ T cells. To study whether these age-related changes were due to previous antigenic exposure, we performed a phenotypic and functional analysis on splenic CD4+ T cells isolated from individual germ-free (GF), specific pathogen-free (SPF), and clean conventional (CC) mice. Interestingly, the total number of splenic CD4+ T cells in GF mice was twofold lower as compared to age-matched SPF or CC mice, regardless whether mice were analyzed at young (10 weeks) or at advanced age (13-14 months). Unexpectedly, the phenotypic composition of the CD4+ T-cell subset was comparable in the GF, SPF, and CC mice as determined by the expression of CD45RB and Pgp-1, indicating that CD4+ T cells with a naive phenotype (CD45RB++ Pgp-1-) were not enriched in GF mice. Moreover, at an age of 13-14 months, CD4+ T cells from GF mice frequently produced more IL-4 and IFN-gamma than their CC counterparts. These lymphokine data showed, therefore, that a relatively high proportion of CD4+ T cells with a memory phenotype can also be defined in GF mice on the basis of their function. The contamination of GF mice with a colonization resistant factor (CRF flora) resulted in twofold higher numbers of splenic CD4+ T cells. Surprisingly, not only CD4+ T cells with a memory phenotype (CD45RB-/+ Pgp-1++) had expanded, but also CD4+ T cells with a naive (CD45RB++ Pgp-1-) phenotype. Our results, therefore, strongly suggest that the expansion of naive CD4+ T cells in the periphery is mediated by the intestinal microflora.


Analysis of immature (CD4-CD8-) thymic subsets in T-cell receptor alpha beta transgenic mice.

Introduction of a transgenic alpha beta TCR (V alpha 2, V beta 8.1) specific for lymphocytic choriomeningitis virus (LCMV), in the context of H-2Db into the genome of C57BL/6 mice, has many effects on the development and selection of T cells in both the thymus and the periphery. These mice produce increased numbers of CD4-8+ mature T cells, all of which express the transgenic TCR, and small numbers of CD4+8- cells using endogenous TCRs are also produced. This study follows the intrathymic development of T cells in these TCR alpha beta transgenic mice, in particular the earliest CD4-8- stages. As expected, the transgenic TCR is expressed on the cell surface at an earlier developmental stage than endogenous TCRs in nontransgenic littermate controls. Of the three major subsets expressing the heat-stable antigen (HSA), only the most mature, the CD25-CD44- expresses the transgenic TCR, and the earlier CD25-CD44+ and CD25+CD44- do not. Furthermore, in contrast to other TCR alpha beta transgenic lines, TCR gamma delta lineage cells appear to develop normally.


Specific activation of platelets from patients allergic to Dermatophagoides pteronyssinus by synthetic peptides derived from the allergen Der p I.

Previous studies have shown that Fc epsilon RII-bearing platelets from patients with allergic disorders could be stimulated either by anti-human IgE antibodies or the relevant allergens to generate cytotoxic mediators. In this report, the reactivity of platelets from Dermatophagoides pteronyssinus (D. pt)-sensitive patients to three Der p I-derived synthetic peptides (52-71, 117-133 and 188-199) was investigated. The platelet stimulation observed in D. pt-sensitive patients was demonstrated to be allergen-specific and to be mediated by IgE. These Der p I-derived peptides failed to stimulate platelets from healthy donors or platelets from non-D. pt-allergic patients. D. pt-sensitive patients responded more frequently to the peptides 52-71 and 117-133 than to the peptide 188-199. Thus, in this model, synthetic peptides selected for their assumed accessibility on the native antigen could be associated to an IgE-dependent biological activity.


Use of immobilized metal ions as a negative adsorbent for purification of enzymes: application to phosphoglycerate mutase from chicken muscle extract and horseradish peroxidase.

Two enzymes, phosphoglycerate mutase and peroxidase, were purified by using an immobilized metal ion adsorbent for the removal of unwanted proteins. The mutase was obtained pure from a single column, whereas the purification of peroxidase required the use of a thiophilic adsorbent in a tandem. The capacity was 2.5 mg pure peroxidase per mL gel.


Prognostic factors for radiographic damage and physical disability in early rheumatoid arthritis. A prospective follow-up study of 147 patients.

We studied the influence of demographic, clinical, laboratory and genetic features and radiographic damage at onset on the outcome after 2 years in a prospective study of 147 patients with classical or definite RA with disease duration shorter than 1 year at entry. Outcome was determined by physical disability and by radiographic damage of hands and feet. By means of multiple regression analysis and discriminant analysis outcome was explained from variables at the start and during the first 6 months. No clinically relevant conclusions could be made for physical disability due to the low explained variance and small number of patients with bad physical disability. Radiographic damage after 2 years was predicted by high disease activity at the start (measured as erythrocyte sedimentation rate, C-reactive protein or Disease Activity Score) combined with DR4 or DR2 (as a prognostically favourable factor) and rheumatoid factor positivity. Radiographic damage could be better predicted if disease activity during the first 6 months was included. Absence or presence of progression of radiographic damage could be correctly predicted in 83% of the patients.


T gamma delta cells and their subsets in blood and synovial fluid from patients with rheumatoid arthritis.

We have determined the distribution of T gamma delta cells in the peripheral blood of 44 patients with rheumatoid arthritis and in 36 healthy controls. In addition, paired blood and synovial fluid samples were obtained from seven patients with RA. The monoclonal antibodies A13, BB3 and Ti gamma A, which are specific for the V delta 1, V delta 2 and V gamma 9 gene products respectively, were used to define T gamma delta subsets. T gamma delta + cells expressed as a percentage of CD3+ lymphocytes were reduced in RA peripheral blood compared with the control group (3.9% +/- 0.5 versus 5.7% +/- 0.7; P less than 0.0001). There was a reduction in the V gamma 9/V delta 2+ subset (from 5.6% +/- 1.2 to 1.7% +/- 0.4) leading to a change in the mean ratio of V delta 2/V delta 1+ cells from 4.3 in normal subjects to 1.1 (P less than 0.002). No statistical difference was observed in T gamma delta cell numbers in synovial fluid compared with the paired blood samples (4.0% +/- 1.1 in blood and 4.4% +/- 1.4 in synovial fluid). Also the distribution of V delta 2+ and V delta 1+ cells was similar in the two compartments and a similar alteration in subset distribution was found in blood and synovial fluid. These findings do not indicate a selective accumulation of a specific T gamma delta subset in RA synovial effusions.


Heteroconjugated antibodies enhance lymphocyte-mediated tumour cell lysis in vitro and in vivo.

Covalent linkage of an antitumour antibody specific for a tumour cell surface antigen to an antilymphocyte antibody specific for the T lymphocyte receptor complex produces a heteroconjugated antibody that can activate and redirect cytotoxic T lymphocytes to lyse tumour cells. The ability of an antilymphocyte-antitumour heteroconjugate (500A2 x 96.5) to direct the lysis of murine melanoma cells by cultured murine lymphocytes was tested in vitro using a 4-h chromium release assay and in vivo with a tumour neutralization assay. In vitro, the addition of heteroconjugated antibody significantly increased tumour lysis by murine C3H/HeN lymphocytes (median specific lysis 82.7 per cent with lymphocytes plus heteroconjugate versus 9.5 per cent for lymphocytes alone, P less than 0.001). In vivo, treatment with heteroconjugated antibody plus lymphocytes significantly reduced the development of pulmonary metastases after intravenous tumour administration (median number of pulmonary metastases 28.5 for combined treatment versus 250 for heteroconjugate or lymphocytes alone, P less than 0.001).


Effect of anti-CD3 antibody on the generation of interleukin-2-activated lymphocytes from tumor tissues of gastrointestinal cancer.

The efficiency of anti-CD3 antibody (OKT3) for adoptive immunotherapy using lymphokine-activated killer (LAK) cells generated from tumor-infiltrating lymphocytes (TIL), regional lymph node lymphocytes (RLNL), and peripheral blood lymphocytes (PBL) was investigated. TIL, RLNL, and PBL derived from 39 patients with gastrointestinal cancers (16 gastric cancers, 17 colorectal cancers, and 6 esophageal cancers) were cultured for 4 weeks with 200 U/ml of recombinant interleukin-2. To one group, solid-phase 10 micrograms/ml OKT3 was added during the initial culture period (day 2 or 4). Cytotoxicity against K562 cells (NK-like activity) and Daudi cells (LAK activity) and the phenotypes of effector cells generated after culturing for 2-3 weeks were studied. Proliferative responses were significantly increased by OKT3 in each type of effector cell (P less than 0.01); in particular, TIL expanded more by OKT3 than PBL and RLNL (P less than 0.01). The population of CD8+ CD11b- cytotoxic T-cells in OKT3-stimulated groups was significantly larger than that in unstimulated groups (P less than 0.01), whereas no differences were observed with CD4+ cells (helper/inducer T-cells) and CD8+ CD11b+ cells (suppressor T-cells). OKT3 enhanced the NK-like activity of TIL and PBL but did not affect their LAK activity. OKT3 suppressed the NK and LAK activity of RLNL. OKT3 stimulation did not significantly enhance the LAK activity, but the authors propose that OKT3 could be an effective addition to adoptive immunotherapy using TIL due to an increased proliferation and generation of a large cytotoxic T-cell population.


Role of endocrine, autocrine, and paracrine interactions in the development of mammary hyperplasia in Wnt-1 transgenic mice.

The Wnt-1 proto-oncogene is transcriptionally activated by mouse mammary tumor virus in mouse mammary tumor virus-induced tumors. Previous studies using transgenic mice showed that Wnt-1 expression in mammary gland causes alveolar hyperplasias which resemble mammary glands of pregnant mice. To understand the role of mammogenic hormones in the genesis of these hyperplasias, we examined the development of these glands before puberty in young transgenic mice and the effects of ovariectomy and adrenalectomy on the growth and morphology of Wnt-1 mammary hyperplasia. Mammary glands of Wnt-1 transgenic females showed hyperplastic morphology as early as 1 week after birth. The normal structure of the uterus of the adult Wnt-1 virgin mouse indicated that the circulating levels of ovarian hormones were not elevated. Ovariectomy and adrenalectomy had no obvious effect on the morphology of these mammary hyperplasias. To assess possible paracrine stimulation of mammary epithelial cells (MEC) by stromal cells, we transplanted MEC from normal BALB/c mice into gland-free fat pads of Wnt-1 transgenic mice and found that normal MEC maintained their normal ductal structure in Wnt-1 fat pads without alveolar development. Further, we did not detect Wnt-1 mRNA expression in the gland-free fat pads of these transgenic mice. When Wnt-1 MEC were transplanted into the fat pads of nude mice and allowed to grow towards existing normal MEC, the morphology of the existing normal MEC remained normal. We concluded that the development of mammary hyperplasia in Wnt-1 transgenic mice is solely dependent on Wnt-1 expression in MEC. We speculate that Wnt-1 may be a growth factor for mammary gland that only acts locally on the cells that produce it.


Role of the alpha 5 beta 1 integrin receptor in the proliferative response of quiescent human melanoma cells to fibronectin.

The possible mitogenic activity of fibronectin (FN) in human primary and metastatic melanoma lines and clones and the involvement of integrins in mediating this effect were evaluated. Quescent human melanoma cells cultured in serum-free medium proliferated in a dose- and time-dependent fashion to immobilized FN as indicated by [3H]thymidine incorporation, increment of cell number, and cell cycle analysis. This response to FN was observed with tumor clones isolated from a subcutaneous metastasis and with primary or metastatic melanomas from different patients, but only when tumor cells expressed the alpha 5 subunit of the FN receptor (i.e., VLA-5). Proliferation to FN by a primary tumor (Me4405) expressing all FN receptors and by a tumor clone (2/60) lacking only the alpha 4 subunit was inhibited by monoclonal antibodies to the alpha 5 and beta 1 but not by monoclonal antibodies to other subunits of FN receptors. Mapping of FN regions responsible for the proliferative signal was performed by stimulating melanoma cells with different FN proteolytic fragments and indicated that a significant mitogenic signal was provided by the M(r) 120,000 alpha-chymotrypsin fragment containing the Arg-Gly-Asp sequence. The proliferation of melanoma cells to FN and to FN fragments was also significantly inhibited by peptides containing the Arg-Gly-Asp sequence. These data indicate that FN can stimulate the proliferation of quiescent melanoma cells and that integrins as alpha 5 beta 1 are involved in the response of tumor cells to this extracellular matrix protein.


Translation of the prophage lambda cl transcript.

Mutations in rpsB that reduce the levels of the ribosomal protein S2 enhance the translation of cl in lambda lysogens. Two features of the cl transcript are required for enhanced translation: the absence of a leader and the presence of a downstream box, a sequence within the cl coding region that is complementary to the 16S rRNA. 30S ribosomal subunits deficient in S2 form ternary complexes with the cl transcript more efficiently than wild-type subunits. The absence of S2 may change the structure of the 16S rRNA, improving contacts with the cl downstream box.


Hepatocyte growth factor and transforming growth factor-beta stimulate both cell growth and migration of human gastric adenocarcinoma cells.

Hepatocyte growth factor (HGF) induced scattering and cell migration of human gastric adenocarcinoma MKN-74. HGF also significantly promoted the growth of MKN-74 cells in a dose-dependent manner, although HGF is reported to be antiproliferative for the growth of tumor cell lines. This result indicates that HGF stimulates cell proliferation of not only normal epithelial cells but also certain carcinoma cells. Furthermore, transforming growth factor-beta (TGF-beta), which is recognized to inhibit the growth of most epithelial cells, additively enhanced both the cell proliferation and migration induced by HGF. These additive effects of HGF and TGF-beta may be responsible for the tumor invasiveness and uncontrolled growth of certain types of carcinoma.


Systemic lidocaine and human somatosensory-evoked potentials during sufentanil-isoflurane anaesthesia.

The effect of systemically administered lidocaine on somatosensory evoked potentials (SSEPs) during general anaesthesia has not been widely reported. Knowledge of the influence of anaesthetic agents on evoked potentials assists in interpreting evoked potential waveforms. Accordingly, we studied the behaviour of cortical and subcortical (recorded at the second cervical vertebra) SSEPs after administration of intravenous lidocaine (3 mg.kg-1 bolus followed by infusion at 4 mg.kg-1.hr-1) during a sufentanil-based anaesthetic regimen in 16 patients undergoing abdominal or orthopaedic surgery. When compared to awake baseline recordings, the sufentanil-nitrous oxide, low-dose isoflurane anaesthetic depressed N1 amplitude by approximately 40% and prolonged latency by 10%. Fifteen minutes after establishment of this anaesthetic, the amplitude and latency of N1 were 1.13 +/- 0.56 microV and 19.81 +/- 1.63 msec, respectively. Within five minutes of adding lidocaine, amplitude decreased further to 0.84 +/- 0.39 microV (P = 0.001), while latency was extended to 20.44 +/- 1.48 msec (P = 0.01). Lidocaine did not affect cervical amplitude and prolonged latency only minimally. Despite the observed effects on amplitude and latency, SSEP waveforms were preserved and interpretable. Plasma lidocaine levels obtained at 5, 20, and 40 minutes after lidocaine were 5.17 +/- 1.33, 3.76 +/- 1.14, and 3.66 +/- 0.9 micrograms.dl-1, respectively. Our results indicate that systemically administered lidocaine at therapeutic plasma levels acts synergistically with a sufentanil-based anaesthetic to depress the amplitude and prolong the latency of SSEPs.


Evaluating health and maturation of the unborn: the role of the clinical laboratory.

I review the utility of several common prenatal laboratory tests. Infant mortality is higher in the United States than in many other industrialized nations; better access to early prenatal care may help reduce this mortality rate. Several common laboratory tests can significantly contribute to prenatal care. Early measurement of maternal serum human chorionic gonadotropin can permit estimation of the date of conception. Use of maternal serum alpha-fetoprotein in the second trimester provides the clinician with risk estimates for neural tube defects and Down syndrome. Adding human chorionic gonadotropin and possibly unconjugated estriol to this screen can increase the number of Down syndrome cases identified without increasing the proportion of abnormal results. Amniotic fluid fetal lung maturity tests can assist with the management of delivery.


On the origin of C3 nephritic factor (antibody to the alternative pathway C3 convertase): evidence for the Adam and Eve concept of autoantibody production.

The antibody to the alternative pathway C3 convertase, designated C3 nephritic factor or C3NeF, is an autoantibody that is produced in everyone from the time of birth. The elaboration of C3NeF utilizes germline V-region genes which undergo antigen-driven affinity maturation, resulting in an autoantibody that is produced in large amounts with high affinity and narrow specificity. Our data also suggest that under normal conditions, the idiotypic network may play an important part in the control of this autoantibody. Further, a defect in the network with loss of control or inappropriate stimulation may be an underlying mechanism in the unrestricted production of C3NeF in patients with membranoproliferative glomerulonephritis.


A modification of the in vivo mixed lymphocyte reaction and rapamycin's effect in this model.

Rapamycin, a novel macrocyclic immunosuppressive agent, suppresses murine T cell activation in vitro by mechanisms distinct from cyclosporin A (CsA). This study was designed to examine rapamycin and CsA in the host vs graft popliteal lymph node (PLN) model, an in vivo system of T cell-dependent lymphocyte activation. The PLN procedure was modified by using irradiated CTLL-2 cells of C57BL/6 origin, instead of primary mouse splenocytes, as the allogeneic stimulus in C3H/HeN recipient mice. PLN cell proliferation was determined by [3H]-thymidine uptake. We found that the host lymphocyte proliferative response to CTLL-2 cells (H-2b) is greater than the response to mouse Balb/c splenocytes (H-2d). Rapamycin (ip or po) produced a dose-related inhibition of the in vivo mixed lymphocyte reaction. By contrast, the effects of CsA and FK-506 were not dose related within the same dose range (0.006-12 mg/kg). These data indicate that rapamycin is an effective immunosuppressive agent and confirm its ability to affect the allogeneic T cell response in vivo. Furthermore, the pharmacological data suggest that this PLN model utilizing irradiated CTLL-2 cells as an allogeneic stimulus provides a reproducible system to examine mixed lymphocyte reactions in vivo.


The bactericidal activity and postantibiotic effect of trospectomycin.

Trospectomycin sulfate (trospectomycin, TRS) is a novel, broad-spectrum, aminocyclitol antibiotic that is being developed clinically for the treatment of upper respiratory tract infections, bacterial vaginosis, pelvic inflammatory disease, and gonorrhea. This study investigated the bactericidal activity (by time-kill kinetics) and the postantibiotic effect (PAE) of TRS. Species-dependent bacteriostatic/bactericidal activity was observed for TRS; the antibiotic was bacteriostatic for Staphylococcus epidermidis, Enterococcus faecalis, and Escherichia coli, and bactericidal for Haemophilus influenzae, Neisseria gonorrhoeae, Moraxella catarrhalis, and Bacteroides fragilis (one of two test strains). When TRS was tested at four times its minimum inhibitory concentration or at a maximum test concentration of 32 micrograms/ml, with a 1-hr exposure period, the following PAE values were recorded: S. epidermidis 30032, 1.8 hr, En. faecalis ATCC 29212, 1.6 hr, E. coli UC 311, 1.5 hr, E. coli UC 9451, 1.5 hr, H. influenzae 30063, greater than 4.0 hr, B. fragilis ATCC 25285, 5.2 hr, and B. fragilis UC 12199, 6.7 hr. The broad-spectrum PAE that was observed for TRS is somewhat unique compared with other antibiotics.


Addition of enoximone to adrenergic agents in the management of severe heart failure.

To assess the hemodynamic effects of the addition of small bolus doses of the phosphodiesterase inhibitor enoximone to adrenergic therapy in patients with severe heart failure. Open label, prospective study. Multidisciplinary department of intensive care in a university academic hospital. Twelve surgical patients after cardiac surgery and ten medical patients with ischemic or dilated cardiomyopathy who had signs of altered tissue perfusion associated with a low cardiac index (less than 2.25 L/min/m2), despite adrenergic therapy. Small iv bolus doses of 0.25 mg/kg of enoximone. This treatment resulted in significant increases in cardiac index and left ventricular stroke work index without significant changes in heart rate or mean arterial pressure. Furthermore, the effects of half this dose (i.e., 0.125 mg/kg), studied in 11 patients, demonstrated a significant drug-induced increase in mean (+/- SD) cardiac index (from 1.58 +/- 0.29 to 1.84 +/- 0.27 L/min/m2, p less than .01) without change in mean arterial pressure (from 74.5 +/- 12.1 to 76.5 +/- 12.6 mm Hg, nonsignificant). Direct iv injections of enoximone can significantly increase the cardiac index in critically ill patients treated by adrenergic agents for severe heart failure. The administration of small doses of enoximone is effective and has minimal effect on arterial pressure.


Acute myopathy during treatment of status asthmaticus with corticosteroids and steroidal muscle relaxants.

Acute myopathy in patients being treated for severe asthma has been recognized with increasing frequency since first being described in 1977. We report three patients treated for status asthmaticus who developed severe generalized weakness. Electrophysiologic studies and muscle biopsy revealed evidence of muscle destruction. Each of these patients was treated with high-dose corticosteroids and underwent prolonged neuromuscular blockade with a steroidal muscle relaxant. A review of the literature revealed 15 similar cases. We postulate that the combined effects of corticosteroids and muscle relaxants on the muscle cell may be responsible for this myopathy. Patients treated with corticosteroids and NMBs should be carefully monitored for the development of myopathy.


Systemic hypersensitivity vasculitis associated with bronchiectasis.

Systemic hypersensitivity vasculitis developed in a 53-year-old man during acute exacerbation of bronchiectasis infected with Pseudomonas aeruginosa. High grade fever, mononeuropathy multiplex, cutaneous vasculitis, and biopsy specimen-proved mesangioproliferative glomerulonephritis with crescent formation and leukocytoclastic vasculitis associated with circulating immune complex occurred. Corticosteroid and cyclophosphamide therapy was effective for vasculitis and bronchiectasis.


Alterations of nucleolar ultrastructure and ribosome biogenesis by actinomycin D. Implications for U3 snRNP function.

We have studied, at the electron microscope level, the reorganizations of nucleolar ultrastructure induced by actinomycin D (AMD) in different conditions of drug treatment associated with an inhibition of rRNA synthesis. We have analyzed in parallel the localizations of ribosomal genes, of their transcripts, of various pre-rRNA intermediates, as well as of U3 RNA and fibrillarin by in situ hybridization with nucleic acid probes and immunocytological detection on thin sections of human and mouse cells. Consistent with previous observations, dense fibrillar component (DFC) and granular component (GC) appear to contain distinct pre-rRNA species at different stages of their processing. DFC appears as a major site of U3 RNA accumulation, but a very substantial fraction of nucleolar U3 RNA is also found in GC, colocalizing with partially processed pre-rRNAs. Remarkably, the major nucleolar components retain their ultrastructural appearance when extensively depleted of their pre-rRNA moiety, and ribosomal genes are always detected over fibrillar center (FC), even after extended AMD treatments which result in the characteristic segregation of nucleolar components. Moreover, while for GC the U3 RNA and pre-rRNA contents evolve in parallel following the cessation of rRNA synthesis, a dramatic uncoupling is observed for DFC. The persistent presence of U3 RNA and fibrillarin after pre-rRNA depletion suggests that DFC could represent an anchorage site for U3 snRNPs, before their entering another cycle of pre-rRNA processing reactions.


Exercise testing at 3 weeks, 6 weeks and 18 months after infarction and the outcome at 3 years in young patients (under 55 years).

To examine the prognostic value of routine postinfarction exercise tests in young patients, exercise tests were carried out at 3 and 6 weeks and 18 months after infarction in 149 patients aged under 55 years at the time of the index infarction. The patients also had coronary angiography and left ventriculography a mean of 3 months after infarction. Three years after infarction, only two of the 149 patients have died, reinfarction occurred in only seven (4.7%) patients; unstable angina in four (3%) patients and coronary artery surgery was needed in 31 (20.8%) patients; 16 in the first, 10 in the second, and 5 in the third year of follow-up. Angina on exercise testing at 6 weeks was the only variable with any predictive value. Eighteen (38%) of the 47 patients with, compared to 12 (11.8%) of the 102 patients without, angina on exercise testing at 6 weeks had coronary surgery (less than 0.001). None of the other exercise variables reliably predicted death, or other complications, including coronary surgery. Ten (13.8%) of the 75 patients excluded from the study died during follow-up; six of them within 6 weeks of infarction. Four (67%) of these patients were excluded from the study because of heart failure. Therefore, the 3-year outcome in young survivors of a myocardial infarction is good and is not reliably predicted by exercise testing at 3 and 6 weeks or 18 months.


Differences in blood pressure regulation of congestive heart failure, before and after treatment, correlate with changes in the circulating pattern of atrial natriuretic peptide.

The mechanisms underlying altered BP regulation in congestive heart failure are unknown. This study examines the possibility that differences in circadian blood pressure (BP) regulation between the normal and the failing heart correlate with changes in the circulating pattern of atrial natriuretic peptide (ANP). Twelve normotensive patients with coronary artery disease were studied over two separate 24-h periods, the first during acute exacerbation of congestive heart failure (radionuclide-determined ejection fraction at rest was less than 30%) and the second after therapy-induced functional recovery (ejection fraction was more than 40%). BP monitoring at 10-min intervals and intra-atrial blood samples for ANP assays at hourly intervals were obtained. Significant correlation between ejection fraction and the indexes of circadian BP variability (standard deviation of the 24-h pressure mean and day-night pressure difference) were found both before and after treatment. Ejection fraction was independent of the BP means (24-h, daytime and night-time). BP variability, 24-h mean and daytime mean were higher after treatment. ANP means were lower after treatment, whereas ANP variability was higher. The indexes of BP and ANP variability correlated both before and after treatment, whereas the BP and the ANP means were independent. These findings demonstrate that differences in BP regulation of CHF before and after effective treatment correlate with changes in the circulating pattern of ANP. We speculate that by modulating ANP release, the heart could be actively involved in BP regulation as part of the compensatory mechanisms aimed at protecting against circulatory overload.


Amplitude analysis of stress technetium-99m methoxy isobutylisonitrile images in coronary artery disease.

To determine the role of rest and stress gated technetium-99m methoxyisobutylisonitrile (sestamibi), in the detection of coronary artery disease, routine Fourier analysis of these images was performed with the best septal left anterior oblique (LAO) position of 20 patients (17 men, 3 women; aged 40-75 years) who also underwent rest or redistribution/stress single photon emission tomography (SPET) (99mTc-sestamibi and Thallium-201), gated blood pool imaging and coronary angiogram. There were 6 patients with single-vessel disease, 6 with two-vessel disease, 4 with three-vessel disease, 2 with coronary spasms, 1 with a patent graft and 1 with anginal episodes but a normal angiogram result. Three normal volunteers (2 women, 1 man; aged 24-26 years) also had rest and stress gated blood pool as well as rest and stress gated 99mTc-sestamibi imaging. Rest and stress 99mTc-sestamibi amplitude and phase images depicted regional myocardial wall shortening from the outer layer of the myocardium to the center of the left ventricle as follows: a high amplitude halo of maximal negative count rate variation; a circular thinner halo of negligible amplitude; a central region of maximal positive count rate variation, as the images evolved from end-diastole to end-systole. Similar patterns with regional differences represented abnormal myocardial wall shortening. 99mTc-sestamibi and 201Tl SPET images were in agreement in 90% of the patients and 92% of myocardial regions. 201Tl SPET detected 83% of angiographically proven lesions, as compared with 80% for 99mTc-setamibi SPET and 80% for the amplitude images. The amplitude images demonstrated a larger number of other abnormalities not predicted on the angiogram, probably because they were able to detect regions with a potential for flow improvement and transient regional wall shortening abnormalities.(ABSTRACT TRUNCATED AT 250 WORDS)


A new principle to normalize plasma concentrations allowing single-sample clearance determinations in both children and adults.

A sufficiently accurate quantification of renal function requiring only one plasma sample without an additional gamma-camera study has, until now, only been possible in adults. A new principle will be presented here allowing the universal application of known algorithms, regardless of the clearance substance used, by normalizing the plasma concentrations with respect to the individual body dimensions of the patients--for infants as well as for adults. In this respect, algorithms are developed for clearance determinations using technetium-99m mercaptoacetyltriglycine (99mTc-MAG3), which are based on steady-state studies as the reference. They allow the calculation of quantitative clearance values in infants, requiring only one blood sampling at any time between the 25th and the 40th min postinjection. The comparison with a combined camera/two-plasma-sample technique performed in 46 children aged between 9 days and 14 years (mean 6.05 years) resulted in a standard error of 8.5% from the line of identity (r = 0.94). Moreover, this procedure also increases the accuracy of results in adults.


The bulbar network of respiratory neurons during apneusis induced by a blockade of NMDA receptors.

Our aim was to study the mechanisms producing the transition from the inspiratory phase to the expiratory phase of the breathing cycle. For this purpose we observed the changes affecting the discharge patterns and excitabilities of the different types of respiratory neurons within the respiratory network in cat medulla, after inducing an apneustic respiration with the N-methyl-D-aspartate (NMDA) antagonist MK-801 given systemically. Respiratory neurons were recorded extracellularly through the central barrel of multibarrelled electrodes, in the ventral respiratory area of pentobarbital-anesthetized, vagotomized, paralyzed and ventilated cats. Inhibitions exerted on each neuron by the pre-synaptic pools of respiratory neurons were revealed when the neuron was depolarized by an iontophoretic application of the excitatory amino-acid analogue quisqualate. Cycle-triggered time histograms of the spontaneous and quisqualate-increased discharge of respiratory neurons were constructed in eupnea and in apneusis induced with MK-801. During apneustic breathing, the activity of the respiratory neuronal network changed throughout the entire respiratory cycle including the post-inspiratory phase, and the peak discharge rates of all types of respiratory neurons, except the late-expiratory type, decreased. During apneusis, the activity of the post-inspiratory neuronal pool, the post-inspiratory depression of other respiratory neurons, and the phrenic nerve after-discharge were reduced (but not totally suppressed), whereas the discharge of some post-inspiratory neurons shifted into the apneustic plateau. The shortened post-inspiration (stage 1 of expiration) altered the organization of the expiratory phase. Late-expiratory neurons (stage 2 of expiration) discharged earlier in expiration and their discharge rate increased. The inspiratory on-switching was functionally unaffected. Early inspiratory neurons of the decrementing type retained a decrementing pattern followed by a reduced discharge rate in the apneustic plateau, whereas early-inspiratory neurons of the constant type maintained a high discharge rate throughout the apneustic plateau. Inspiratory augmenting neurons, late-inspiratory and "off-switch" neurons also discharged throughout the apneustic plateau. During the apneustic plateau, the level of activity was constant in the phrenic nerve and in inspiratory neurons of the early-constant, augmenting, and late types. However, progressive changes in the activity of other neuronal types demonstrated the evolving state of the respiratory network in the plateau phase. There was a slowed but continued decrease of the activity of early-inspiratory decrementing neurons, accompanied by an increasing activity and/or excitability of "off-switch", post-inspiratory and late-expiratory neurons. In apneusis there was a decoupling of the duration of inspiration and expiration.(ABSTRACT TRUNCATED AT 400 WORDS)


Donor alloreactivity may predict acute graft-versus-host disease in HLA-matched bone marrow transplantation for leukemia in early remission.

The pretransplant alloantigen-dependent responding and stimulating capacity of donors and of recipients was studied retrospectively, in a study of prediction of acute graft-versus-host disease. Donor responding capacity (DRC) and host stimulating capacity (HSC) were defined by mixed lymphocyte culture (MLC) and normalized by help of the pool response. High and low DRC and strong and weak HSC was defined by distribution plots. Kaplan-Meier estimates of the risk of developing Grade II or higher aGvHD showed that first remission patients (N = 125) had a significantly different risk if transplanted with marrow from a donor with high (N = 54) or low (N = 71) DRC (chi 2 = 9.49; d.f. = 1; p less than 0.002). This was not the case for patients transplanted in later remissions. Host SC status had no significant influence on the aGvHD status (chi 2 = 1.75 and 2.40; d.f. = 1; p = 0.19 and 0.12, defined by normal controls A and B respectively). In conclusion, the results indicate that pretransplant donor alloreactivity may predict aGvHD, confirming the results from a Scandinavian study. The results need to be confirmed in prospective studies of alloreactivity as risk factor for aGvHD.


Regulation of neuronal migration and neuritogenesis by distinct surface proteases. Relative contribution of plasmin and a thrombin-like protease.

The relative contribution of two neuronal surface proteases, plasmin and a protease with thrombin-like specificity, on NB2a/dl neuroblastoma migration and neuritogenesis were examined. Exogenous plasmin induced cell body rounding and increased cell migration, but did not prevent or reverse neurite outgrowth. Inhibition of endogenous plasmin by its specific inhibitor, aprotinin, suppressed migration but did not induce neuritogenesis. Removal or inhibition of the thrombin-like protease by serum deprivation or hirudin addition, respectively, induced neurite outgrowth, as shown in our previous studies, but did not suppress migration. By contrast, trypsin induced simultaneous cell rounding and neurite retraction. These findings indicated that plasmin may regulate cell migration, while the thrombin-like protease may regulate facets of neurite outgrowth. Although unable to induce de novo neuritogenesis, plasmin inhibition potentiated the otherwise transient neurites induced by simultaneous inhibition of the thrombin-like protease. Since cultured neuronal cells migrate primarily in the direction of newly elaborated neurites, this finding is interpreted to indicate that cessation of neuronal migration by plasmin inhibition enhances net neurite outgrowth by inhibition of the putative thrombin-like protease.


In vitro interaction of fenretinide with plasma retinol-binding protein and its functional consequences.

The synthetic retinoid fenretinide (4-HPR; N-[4-hydroxyphenyl] all-trans-retinamide) interacts with plasma apo-retinol-binding protein (RBP) to form a tight complex (K'd approximately 0.2 microM) which does not exhibit binding affinity to transthyretin (TTR). Therefore, a substantial modification of the retinol hydroxyl group does not appear to affect the interaction with RBP but does drastically interfere with the protein-protein recognition. The remarkable early reduction in plasma retinol level induced by fenretinide administration may be associated with the high binding affinity of this retinoid to RBP and to its interference with the RBP-TTR complex formation.


The metastatic potential of rat prostate tumor variant R3327-MatLyLu is correlated with an increased activity of N-acetylglucosaminyl transferase III and V.

Enzyme activities of N-acetylglucosaminyltransferase (GlcNAc-Tase) I-V involved in N-linked complex-type carbohydrate synthesis were determined in a non-metastatic hormone-dependent rat prostate tumor (R3327-H) and a related, hormone-independent variant metastasizing to lymph nodes and lungs (R3327-MatLyLu). In the metastasizing variant a significantly increased activity of both GlcNAc-Tase III and GlcNAc-Tase V was observed, whereas the activities of GlcNAc-Tase I and II were essentially unchanged. The increase in activity of GlcNAc-Tase III is particularly noteworthy since it indicates that elevated expression of this enzyme cannot be considered as an exclusive marker of hepatic malignancy.


Purification and characterization of phosphofructokinase in bovine parotid gland.

1. Phosphofructokinase (PFK) was purified from bovine parotid gland to 750-fold with the specific activity of 67.5 units/mg protein by Cibacron Blue F3GA affinity chromatography, and TSK DEAE-5PW ion-exchange and TSK G4000SW size exclusion chromatographies on HPLC. 2. On gel-filtration, molecular weight of the native PFK was estimated to 400,000. 3. PFK was a heterotetramer composed of three kinds of subunit with molecular weights of 92,000 (C-type), 88,000 (M-type) and 86,000 (L-type), by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Densitometrically, relative amounts of C-, M- and L-type subunit were 1:1:2. 4. Under the physiological conditions of fructose 6-phosphate (Fru-6-P) and ATP concentrations and pH, PFK activity was suppressed and hardly detectable. 5. Fru-6-P relieved PFK from the ATP inhibition. 6. Fructose 2,6-bisphosphate (Fru-2,6-P2) and AMP activated PFK with a reduction of S0.5 for Fru-6-P and subunit cooperativity. Fru-2,6-P2 was more effective than AMP.


Timing of synthesis and cellular localization of two conidiation-specific proteins of Neurospora crassa.

The process of conidiation in Neurospora crassa consists of a series of distinct developmental stages culminating in the formation of multinucleate asexual spores called macroconidia. Immunoblotting techniques were used to study the timing of synthesis and cellular localization of CON10 and CON13, the products of two genes that are expressed during conidiation but not during mycelial growth. Both proteins first appear about 8 hr into conidiation; CON10 disappears between 2 and 4 hr after germination. Within conidiating cultures, CON10 and CON13 proteins are localized in conidiophores, with little or no protein present in the underlying mycelium. Immunofluorescence analyses show that CON10 is evenly distributed throughout the cytoplasm of macroconidia. Synthesis of CON10 and CON13 occurs at a time when their specifying mRNAs first appear (Hager and Yanofsky, Gene 96, 153-159, 1990; Sachs and Yanofsky, Dev. Biol 148, 117-128, 1991), suggesting that regulation of synthesis is predominantly transcriptional.


Communication aids for children: procedures and problems.

This paper describes the organisation and procedures of the Communication Aids Centre for children at the Wolfson Centre, London, including a model for assessment and recommendation of appropriate aids, such as symbol charts, switches and speech synthesisers. Of the children seen over an 18-month period, most had cerebral palsy and two-thirds were wheelchair-dependent. Almost half were assessed before the age of five years. A detailed follow-up of nine children is presented which reveals how long children may have to wait for the provision of an aid in the UK. Possible problems in establishing use of an aid are discussed; these include inadequate training of children and their communication partners. Suggestions for future improvements of communication-aids services are explored.


The heart and control of renal excretion: neural and endocrine mechanisms.

There has been a great deal of research concerning the heart being an important regulator of renal fluid and electrolyte excretion. This cardiac-renal connection involves two different types of major mechanisms, both of which are covered in this review. The first of these to be discovered was neural reflex regulation. This type of control is due to the fact that the heart possesses nerve receptors whose activity is altered by changes in the degree of cardiac stretch that occur as a result of changes in blood volume. These receptors affect various humoral, neural, and perhaps hemodynamic mechanisms that modify renal excretion. A second, more recently discovered type of regulation is based on the concept that the heart is also an endocrine gland. Similar to neural receptor activity, cardiac hormone secretion is also linked to the degree of cardiac stretch or filling. These cardiac peptides have been shown to have a variety of physiologic effects, most of which directly or indirectly affect renal excretion. Both of the above cardiorenal control mechanisms, one neural and one humoral, may be important not only in maintaining normal fluid-electrolyte balance but may also have pathophysiologic relevance.


Therapeutic potential of modulating potassium currents in the diseased myocardium.

Myocardial disease states are characterized by multiple electrophysiologic abnormalities, including alterations in potassium channel activities. During acute myocardial ischemia, activation of ATP-regulated K+ current (IK(ATP)) results in shortening of action potential duration and elevation of extracellular K+ concentration. In hypertrophied myocardium, increases in inward rectifier K+ current (IK1) and decreases in delayed rectifier K+ current (IK) are observed. Alterations in K+ channel activity in myocardial disease states suggest the potential to therapeutically modify cardiac rhythm and function with K+ channel modulators. Class III anti-arrhythmic agents, which prolong myocardial refractoriness predominantly via a blockade of IK, have demonstrated efficacy in suppressing reentrant atrial and ventricular arrhythmias in animal models as well as promising efficacy in initial clinical studies. Potassium channel openers (PCOs), which activate cardiac IK(ATP), have demonstrated both antiarrhythmic and proarrhythmic activities in various experimental settings, and also are being investigated as potential cardioprotective agents. Sulfonylureas, which block cardiac IK(ATP), also have been investigated as potential antiarrhythmic agents with equivocal results, and have displayed a propensity to exacerbate ischemic myocardial dysfunction in experimental studies. A more comprehensive understanding of K+ channel activity in various myocardial disease states, including concomitant disorders such as myocardial ischemia and hypertrophy, will facilitate the development of more useful potassium channel modulators, as well as a clearer recognition of the undesirable effects of such agents.


Subchronic toxicity study of Caramel Colour II in F344 rats.

Caramel Colour II is a distinct type of colourant with a pronounced reddish hue. It is made with sulphite reactants but without ammonia. The red colour and a high alcohol solubility provide functional characteristics that are important in foods or beverages containing natural flavour extractives. Caramel Colour II is widely used in ice creams and liqueurs; however, it represents less than 1% of total caramel colour manufacture. The toxicity of Caramel Colour II was evaluated in a 13-wk study in Fischer-344 (F344) rats. The test material was mixed with demineralized water and the solutions were given to the animals ad lib. in the drinking fluid. The concentrations of caramel colour in the drinking fluid were adjusted periodically to achieve the desired caramel colour intake/kg body weight/day. Groups of 20 rats/sex were given Caramel Colour II at levels of 0, 4, 8, 12 or 16 g/kg for at least 13 wk. There were no deaths in any of the groups fed Caramel Colour II. All rats fed caramel colour had soft faeces. All treated groups also had lower fluid consumption that was attributed to poor palatability of the high concentrations of caramel colour that were fed. A number of changes observed (reduced food consumption in all treatment groups except males given 4 g/kg; significantly lower body weights for males given 12 g/kg or more and for females given 8 g/kg or more; lower urine volume and higher specific gravity) were attributed to the reduced water intake and not considered to be toxicologically significant. There were no consistent treatment-related alterations in haematology or blood chemistry variables, and random changes noted were not associated with macroscopic or microscopic pathological alterations. There were no toxicologically important pathological findings. Based on this study, Caramel Colour II was not toxic in F344 rats treated for 13 wk. The highest dose level tested in this study (16 g/kg) was considered to be the no-observed-adverse-effect level.


A novel mitochondrial genome organization for the blue mussel, Mytilus edulis.

The sequence of 13.9 kilobases (kb) of the 17.1-kb mitochondrial genome of Mytilus edulis has been determined, and the arrangement of all genes has been deduced. Mytilus mitochondrial DNA (mtDNA) contains 37 genes, all of which are transcribed from the same DNA strand. The gene content of Mytilus is typically metazoan in that it includes genes for large and small ribosomal RNAs, for a complete set of transfer RNAs and for 12 proteins. The protein genes encode the cytochrome b apoenzyme, cytochrome c oxidase (CO) subunits I-III, NADH dehydrogenase (ND) subunits 1-6 and 4L, and ATP synthetase (ATPase) subunit 6. No gene for ATPase subunit 8 could be found. The reading frames for the ND1, COI, and COIII genes contain long extensions relative to those genes in other metazoan mtDNAs. There are 23 tRNA genes, one more than previously found in any metazoan mtDNA. The additional tRNA appears to specify methionine, making Mytilus mtDNA unique in having two tRNA(Met) genes. Five lengthy unassigned intergenic sequences are present, four of which vary in length from 79 to 119 nucleotides and the largest of which is 1.2 kb. The base compositions of these are unremarkable and do not differ significantly from that of the remainder of the mtDNA. The arrangement of genes in Mytilus mtDNA is remarkably unlike that found in any other known metazoan mtDNA.


[Eligibility for intensive nursing care. Part 1: Guidelines and current experience].

Against the background of the planned statutory Pflegeversicherung (insurance for nursing care) in Germany, a report is presented on experience gained to date with the new Leistungsbestand "Schwerpflegebedürftigkeit" (situation requiring social benefit "urgent need of nursing care") (Para. 53 ff. SGB V). The experience gained so far is of particular interest since, for the first time on a large scale, monetäre Erstattungsleistung (the provision of a monetary entitlement) replaces the Sachleistungsprinzip (benefit in the form of services) that is otherwise applied in the area of health insurance. It is intended that this form of entitlement will form the core of the planned nursing care insurance scheme.


[Ondansetron--the first highly selective 5-HT3 antagonist in therapy of psychiatric diseases].

Ondansetron is a highly selective 5-HT3 antagonist, which has recently become available for the control of chemotherapy-induced emesis. Since 5-HT3 receptors not only have a high density in the area postrema but also in the hippocampal and amygdala region of the limbic system, it has been suspected that 5-HT3 selective agents have psychotropic effects. In animal models of anxiety ondansetron showed a benzodiazepine-like anxiolytic effect without any sedation or withdrawal effects. Other states of withdrawal have been prevented with ondansetron. This agent might also exert neuroleptic effects since dopaminergic hyperactivity in the mesolimbic system was antagonised by ondansetron. In different models of memory and learning a positive effect on basal learning behaviour and on scopolamine-induced memory impairment was noted. This manuscript reviews essential pharmacological and behavioural effects of ondansetron as well as preliminary data from clinical studies. The role of highly-selective ligands for a more differentiated view of serotonergic subsystems are discussed.


[Negative appendectomies can be decreased by improved clinical assessment alone].

We had a 20% rate of negative appendectomies in our patients presenting with suspected appendicitis. We suggested that an improved clinical examination would reduce this rate. 84 consecutive patients presenting with suspected acute appendicitis were prospectively studied. 10 clinical features were used to calculate a score which should distinguish appendicitis and non-specific abdominal pain. 53 appendectomies with 6 (11.3%) perforations, 41 (77.4%) acute inflammations and 6 (11.3%) normal appendixes have been performed. 26 patients suffered from non-specific abdominal pain, 5 had an other surgical disease. In the appendectomized patients the score was 4.2 +/- 1.2 with perforation, 4.4 +/- 1.1 with acute inflammation and 3.8 +/- 1.3 with a normal appendix (p = ns). The score for non-specific abdominal pain in patients without operation was significantly lower (2.0 +/- 1.1; p less than 0.01). Patients with other surgical disease had a score of 2.8 +/- 1.5 with no significant difference to patients which had undergone appendectomy. Negative appendectomies were reduced by improved clinical examination from 20.3% to 11.3% without change in the rate of perforation. The remaining patients with negative appendectomies could not be identified by improved clinical examination even by means of the score. But the use of the score improved the performance of the clinicians.


In vitro activity of clarithromycin and its 14-hydroxy-metabolite against 203 strains of Haemophilus influenzae.

The in vitro activity of clarithromycin alone and in combination with its primary human metabolite, 14-hydroxy-clarithromycin, was determined against 203 strains of Haemophilus influenzae. Microdilution broth MICs and MBCs of both clarithromycin and 14-hydroxy-clarithromycin were determined. The clarithromycin MIC50 was 4 mg/l and the MIC90 was 8 mg/l. The hydroxy metabolite was 2-4-fold more active with an MIC50 and MIC90 of 2 mg/l. The MBCs were equal to the MICs. The microbicidal effect of combinations of clarithromycin and 14-hydroxy-clarithromycin was tested using a microdilution checkerboard technique and the fractional inhibitory index was calculated. The combination was additive in 92% and synergistic in 8% of all strains of H. influenzae tested; no antagonism was found. The results were independent of the site of isolation of the strain or presence of beta-lactamase. These findings suggest the potential clinical utility of clarithromycin for the treatment of H. influenzae infections.


[Appendectomy: open or laparoscopic?].

Surgeons interest in laparoscopic surgery has grown considerably in the last years, mostly due to the explosive spread of laparoscopic cholecystectomy. Laparoscopic appendectomy is from a historical point of view the older procedure, since it was already performed 1982 by Semm. His technique was modified 1987 by Götz, who is also responsible for popularizing it in general surgery. Between April 1989 and April 1991 we have performed 107 laparoscopic appendectomies. 82 were completed successfully by laparoscopy, in 25 instances we had to convert to an open procedure, mostly as a result of the lack of proficiency in the learning period. There were 8 septic complication (9.7%), a rather high rate, which probably will be reduced with the increasing operative expertise. There were no deaths in this series. Further experience is demanded in order to establish laparoscopic appendectomy as the alternative to the conventional procedure.


Risk factors in wound infection following urologic operations: a prospective study.

A total of 134 urologic operations were studied prospectively for postoperative wound infection, the methodology involving direct intraoperative swab taking. Patients' variables were (mean +/- SD): age 32.4 +/- 20.7 years, Quetelet index 27.4 +/- 8, duration of operation 98 +/- 34 minutes, and male:female ratio 9.3:1. Of the 131 intraoperative swabs 28 (21%) were positive, 97% of the organisms being aerobic; 16% of the patients were nasal carriers of S. aureus. The overall wound infection rate was 9%, and it prolonged hospital stay by six days average. Significant risk factors (and their magnitude) were: age over 60 years (x 2.2), prolonged preoperative hospital stay (x 15), and wound contamination (x 4.3 and x 14.3 for classes 3 and 4 wounds respectively). Neither diabetes mellitus, obesity, nor surgeon's rank was contributory. We conclude that, although the 9% rate of postoperative wound infection was acceptable, appropriate prophylactic antibiotics may reduce it further, and, from our data, we would recommend an aminoglycoside (e.g. Amikacin) and Ampicillin combined.


[Is routine intra-operative cholangiography in laparoscopic cholecystectomy truly unnecessary?].

The introduction of laparoscopic cholecystectomy as method of choice for gall stone treatment reopened the question whether to continue with routine intraoperative cholangiography or to switch over to a selective indication. In order to set an accurate indication for selective intraoperative cholangiography it was our goal to develop a tool for preoperative identification of patients with a high risk of common bile duct stones. A preoperative score, indicating the risk of common bile duct stones, was designed. A history of jaundice, elevated levels of bilirubin, alkaline phosphatase, amylase (serum), ALAT (GPT) or ASAT (GOT), a common bile duct wider than 10 mm or containing concrements and multiple gallstones smaller than 10 mm were valued as risk indicators, whereas normal wide bile duct, large or solitary gallstones were valued as decreasing the risk of common bile duct stones. The retrospective screening of 289 consecutive conventional cholecystectomies (1986-1990) for these risk indicators demonstrated a good correlation of the risk score with the occurrence of common bile duct stones. A prospective application of the score, with improved ultrasound examination and routine preoperative intravenous cholangiography, mandatory for laparoscopic cholecystectomy at our institution, will define the high risk group definitely and allow an accurate selective use of intraoperative cholangiography.


The epidemiology of environmental tobacco smoke (ETS) and the weight of evidence argument.

This paper demonstrates that Wald's meta-analysis is an unadjusted combination of unadjusted estimates. There are many differences among the design and implementation of the original studies combined here. One consequence of these differences is that studies of spousal smoking and lung cancer mortality conducted in the United States have a lower combined RR than those done elsewhere. Here it is shown that adjustment for this stratification results is a non significant finding as to the association of ETS with lung cancer. Likewise, the addition, for illustration, of Varela's Ph.D. thesis that is the largest case-control study to date, changes the overall results from a significant to a non-significant result. This paper does not attempt to update Wald's meta-analysis with more recent studies. This is being done, I believe by the Environmental Protection Agency. I have given here my reasons for suspecting the conclusions of meta-analysis of non-randomized studies and the illustrations provided here of the effect of publication bias and covariate adjustment support this view.


[The gallbladder with severe pathologic changes: a contraindication for laparoscopic cholecystectomy?].

In the beginnings of laparoscopic cholecystectomy a severe pathological alteration of the gallbladder or stones of the common bile duct were regarded as relative contraindications to the method. However increasing experience and improve technic have shown, that even a severe pathology of the gallbladder such as chronic cholecystitis with wall thickening, acute or subacute inflammation or a porcelaine gallbladder can be laparoscopically managed. Operation time in such cases is longer, but median hospital stay is the same as in uncomplicated cases. However postoperative morbidity may be increased. For patients with CBD stones preoperative ERCP with papillotomy followed by laparoscopic cholecystectomy some days later offers a treatment with low morbidity and optimal comfort for the patient.


Abdominal colpopexy for complete prolapse of the vagina.

A comparative study of abdominal colpopexy using rectus fascia and sacral fixation for the treatment of prolapsed vagina following hysterectomy was performed. The abdominal approach yields better results than the vaginal route. Fixation of the vaginal vault using a Dacron prosthesis is more rational and was the method of choice, resulting in a complete cure in 10 of 11 patients. The Brady technique failed in 25% of cases.


A reevaluation of estimates of child therapy effectiveness.

In a recent review of nonbehavioral child and adolescent psychotherapy, it was concluded that the magnitude of methodological flaws in this body of research precluded an evaluation of effectiveness. However, recent quantitative reviews of child and adolescent therapy have concluded that nonbehavioral therapy is relatively ineffective compared with behavioral therapy. A sample of nonbehavioral studies was reviewed to evaluate the adequacy of estimates of effectiveness. Three contributions to inaccurate effect estimation were considered: methodological quality, investigator allegiance effects, and treatment representativeness. It was concluded that all three factors contribute to inaccurate estimates of the effectiveness of child and adolescent psychotherapy. Recommendations for improving child therapy research are considered in light of these revealed problems.


The NH2 terminus of retinal recoverin is acylated by a small family of fatty acids.

Recoverin is a recently identified Ca(2+)-binding protein that imparts Ca2+ sensitivity to vertebrate photoreceptor guanylate cyclase. In response to photo-induced depletion of intracellular cGMP and Ca2+, recoverin stimulates resynthesis of cGMP. Bovine retinal recoverin has now been analyzed by electrospray mass spectrometry (ESI-MS) for post-translational modifications that might influence its activity. Heterogeneous acylation was detected at the NH2 terminus of bovine retinal recoverin. The NH2-terminal glycine of each retinal recoverin molecule is linked to one of four different types of acyl groups. The most abundant is myristoleate (14:1), but 14:0, 14:2, and 12:0 acyl residues are also present.


Characterization of autoantibodies that recognize U4/U6 small ribonucleoprotein particles in serum from a patient with primary Sjögren's syndrome.

We identified autoantibodies that recognize the U4/U6 snRNPs in a serum from a 63-year-old Japanese patient (TT) with primary Sjögren's syndrome. This patient's serum immunoprecipitated U4 and U6 sn-RNAs exclusively from 32P-labeled HeLa cell extracts and a newly identified 120-kDa protein along with the Sm core proteins (B'/B, D, E, F, and G) from [35S] methionine-labeled HeLa cell extracts. Immunoblotting demonstrated that only the 120-kDa protein was recognized by this unique serum. In glycerol density gradient centrifugation, the 120-kDa protein reactive with TT serum cosedimented with U4 and U6 snRNAs, suggesting that the 120-kDa protein is a unique component of the U4/U6 snRNP particle. In the same study, the U4/U6 snRNP precipitated by TT serum sedimented only in the lower density, whereas anti-Sm antibodies precipitated U4/U6 snRNAs in a broad range of the gradient. This result suggests the presence of at least two molecular forms of the U4/U6 snRNP particles; larger particles, probably the U4/U5/U6 snRNP complex, and free particles. Thus, the U4/U6 snRNP recognized by TT serum includes the U4 and U6 snRNAs, with Sm core proteins, and the novel 120-kDa protein, and appears to be a free particle not associated with larger complexes.