Patient satisfaction in an outpatient parenteral antimicrobial therapy (OPAT) unit practising predominantly self-administration of antibiotics with elastomeric pumps.

Self-administration of antibiotics using elastomeric pumps has become the most frequently used treatment modality at the outpatient parenteral antimicrobial therapy (OPAT) unit of the University Hospital of Lausanne. However, it remains unknown how comfortable patients feel using this mode of treatment. A questionnaire was offered to all patients treated at the OPAT unit between June 2014 and December 2015. The questionnaire was distributed to 188 patients and 112 questionnaires were returned. Seventy-one patients were treated by self-administration, 21 attended the OPAT unit on a daily basis, and 20 received their antibiotics from home-care nurses. Overall, 83-97% of the patients gave the highest possible scores to the four items evaluating their global satisfaction. Subjects treated by self-administration gave a significantly better rating to 6 of the 17 semi-quantitative questions than the patients treated at the OPAT unit or by home-care nurses. There was no item which was more poorly rated by patients treated by self-administered OPAT than by the other treatment groups. In conclusion satisfaction was high in all patients treated by OPAT. The particularly high satisfaction of patients treated by self-administration of antibiotics with elastomeric pumps suggests that a significant number of patients are happy to take over some responsibility for their treatment. Patients' capacity to appropriate their care themselves should not be underestimated by health care professionals.

Setting up an outpatient parenteral antimicrobial therapy (OPAT) unit in Switzerland: review of the first 18 months of activity.

Outpatient parenteral antimicrobial therapy (OPAT) has been recognised as a useful, cost-effective and safe alternative to inpatient treatment, but no formal OPAT unit existed in Switzerland until recently. In December 2013 an OPAT unit was established at Lausanne University Hospital. We review here the experience of this new OPAT unit after 18 months of activity. Patient characteristics, clinical activities and outcomes were recorded prospectively. Need and acceptance was evaluated as number of OPAT courses administered and number of patients refusing OPAT. Safety and efficacy were evaluated as: (1) adverse events linked to antimicrobials and catheters, (2) re-admission to hospital, (3) rate of treatment failures and (4) mortality. Over 18 months, 179 courses of OPAT were administered. Acceptance was high with only four patients refusing OPAT. Urinary tract infections with resistant bacteria and musculoskeletal infections were the most common diagnoses. Self-administration of antibiotics using elastomeric pumps became rapidly the most frequently used approach. Sixteen patients presented with adverse events linked to antimicrobials and catheters. OPAT-related readmissions occurred in nine patients. The overall cure rate was 94 %. This study shows that OPAT is very well accepted by patients and medical staff, even in a setting which has not used this type of treatment approach until now. Self-administration using elastomeric pumps proved to be particularly useful, safe and efficient. OPAT offers a good alternative to hospitalisation for patients presenting with infections due to resistant bacteria that cannot be treated orally anymore and for difficult to treat infections.

Protein kinase CK2 in health and disease: Cellular functions of protein kinase CK2: a dynamic affair.

Protein kinase CK2 targets a vast array of substrates located in a number of cellular compartments, making the challenge of discriminating among these substrates a daunting task. However, as a signaling protein, CK2 could be targeted to different cellular compartments in response to various stress stimuli such as heat shock, UV irradiation, hypoxia, DNA damage and viral infections. This review will be focused on the evidence that the dynamic association of CK2 subunits and the substrate-dependent subcellular targeting of the enzyme are a likely point of regulation in response to a variety of signaling events. We propose that in addition to enzymatic substrate recognition, regulated CK2 localization to specific compartments should help to provide the exquisite specificity required for robust signal transduction.

Feasibility of two-dimensional ultrasound shear wave elastography of human fetal lungs and liver: A pilot study.

PURPOSE: The first aim was to evaluate feasibility and reproducibility of 2-dimensional ultrasound (2D) shear wave elastography (SWE) of human fetal lungs and liver between 24 and 34weeks of gestation. The second aim was to model fetal lung-to-liver elastography ratio (LLE ratio) and to assess its variations according to gestational age and maternal administration of corticosteroids. MATERIAL AND METHODS: 2D-SWE examinations were prospectively performed in fetuses of women with an uncomplicated pregnancy (group 1) and fetuses of women with a threatened preterm labor requiring administration of corticosteroids (group 2). Two 2D-SWE examinations were performed at "day 0" and "day 2" in group 1; before and 24hours after a course of corticosteroid in group 2. Three operators performed 2 cycles of 3 measurements on the lung (regions A1, A2, A3) and the liver (regions IV, V, VI). Repeatability and reproducibility of measurements were calculated. The fetal LLE ratio was modeled from the most reproducible regions. RESULTS: Fifty-five women were enrolled in group 1 and 48 in group 2. For the lung, 8.6% of measurements were considered invalid and 6.9% for the liver. The most reproducible region for the lung was A3 [ICC between 0.70 (95% CI: 0.42-0.85) and 0.78 (95% CI: 0.48-0.90)] and region VI for the liver [ICC between 0.70 (95% CI: 0.40-0.85) and 0.84 (95% CI: 0.60-0.94)]. According to gestational age, a moderate positive linear correlation was found for stiffness values of A3 (R=0.56), V (R=0.46) and VI (R=0.44). LLE ratio values at "day 0" were not different between the two groups but decreased at "day 2" in group 2 (0.2; 95% CI: 0.07-0.34; P<0.001). CONCLUSION: Quantitative fetal lung and liver stiffness measurements are possible with 2D-SWE with acceptable reproducibility.

p53-dependent inhibition of mammalian cell survival by a genetically selected peptide aptamer that targets the regulatory subunit of protein kinase CK2.

Based on the perturbation of its expression in human cancers and on its involvement in transformation and tumorigenesis, protein kinase CK2 has recently attracted attention as a potential therapeutic target. To assess the value of CK2 as a target for antiproliferative strategies, we have initiated a program aiming to develop inhibitors targeting specifically the regulatory CK2beta subunit. Here, we use a two-hybrid approach to isolate from combinatorial libraries, peptide aptamers that specifically interact with CK2beta. One of these (P1), which has significant sequence homology to the cytomegalovirus IE2 protein, binds with high affinity to the N-terminal domain of CK2beta without disrupting the formation of the CK2 holoenzyme. Expression of green fluorescent protein (GFP)-P1 in different mammalian cell lines activates p53 phosphorylation on serine 15, induces an upregulation of p21 and the release of the Cyt-C and apoptosis-inducing factor proapoptotic proteins triggering caspase-dependent and caspase-independent apoptosis. GFP-P1-induced apoptosis is associated with a p53-dependent pathway as cell death was abrogated in p53 knocked out cells. In summary, our data show that genetically selected peptide aptamers that specifically target CK2beta can induce apoptosis in mammalian cells through the recruitment of a p53-dependent apoptosis pathway. They also emphasize the critical role of CK2beta for cell survival and might allow the design of novel proapoptotic agents targeting this protein.

A site of tyrosine phosphorylation in the C terminus of the epidermal growth factor receptor is required to activate phospholipase C.

Cells expressing mutant epidermal growth factor (EGF) receptors have been used to study mechanisms through which EGF increases phospholipase C (PLC) activity. C-terminal truncation mutant EGF receptors are markedly impaired in their ability to increase inositol phosphate formation compared with wild-type EGF receptors. Mutation of the single tyrosine self-phosphorylation site at residue 992 to phenylalanine in an EGF receptor truncated at residue 1000 abolished the ability of EGF to increase inositol phosphate formation. C-terminal deletion mutant receptors that are impaired in their ability to increase inositol phosphate formation effectively phosphorylate PLC-gamma at the same tyrosine residues as do wild-type EGF receptors. EGF enhances PLC-gamma association with wild-type EGF receptors but not with mutant receptors lacking sites of tyrosine phosphorylation. These results indicate that formation of a complex between self-phosphorylated EGF receptors and PLC-gamma is necessary for enzyme activation in vivo. We propose that both binding of PLC-gamma to activated EGF receptors and tyrosine phosphorylation of the enzyme are necessary to elicit biological responses. Kinase-active EGF receptors lacking sites of tyrosine phosphorylation are unable to signal increased inositol phosphate formation and increases in cytosolic Ca2+ concentration.

A structural basis for substrate specificities of protein Ser/Thr kinases: primary sequence preference of casein kinases I and II, NIMA, phosphorylase kinase, calmodulin-dependent kinase II, CDK5, and Erk1.

We have developed a method to study the primary sequence specificities of protein kinases by using an oriented degenerate peptide library. We report here the substrate specificities of eight protein Ser/Thr kinases. All of the kinases studied selected distinct optimal substrates. The identified substrate specificities of these kinases, together with known crystal structures of protein kinase A, CDK2, Erk2, twitchin, and casein kinase I, provide a structural basis for the substrate recognition of protein Ser/Thr kinases. In particular, the specific selection of amino acids at the +1 and -3 positions to the substrate serine/threonine can be rationalized on the basis of sequences of protein kinases. The identification of optimal peptide substrates of CDK5, casein kinases I and II, NIMA, calmodulin-dependent kinases, Erk1, and phosphorylase kinase makes it possible to predict the potential in vivo targets of these kinases.

Characterization of a cyclic nucleotide-independent protein kinase highly active in human adrenocortical carcinoma.

Study of the protein kinase activity pattern of four human adrenocortical carcinoma showed that in all the samples examined a histone kinase (HK III) activity was present at high level, whereas it was barely detectable in normal tissue. HK III was separated from other known adrenocortical protein kinases by diethylaminoethyl cellulose chromatography. Isolated HK III exhibited a histone (H2B) protamine-phosphotranferase selectivity and used adenosine triphosphate but not guanosine triphosphate as phosphate donor. Serine was identified as the only target amino acid phosphorylated in the protein substrate. HK III showed an apparent molecular weight of 65,000 upon gel filtration and an apparent sedimentation coefficient of 3.7S. HK III activity was cyclic adenosine 3':5'-monophosphate independent and was not influenced by calcium, calmodulin, polyamines, and heparin. The significance of HK III activity in adrenocortical carcinoma extracts at a high level as compared to that of normal tissue remains to be clarified with regard both to its possible relationship with tumoral cell growth and differentiation processes and to its potential interest as a marker of human tumoral tissue activity.

Glucocorticoid binding in the chicken liver cytosol. Characterization of five macromolecular binding components.

The heterogeneity of the glucocorticoid binding in the chicken liver cytosol, previously suggested by the results obtained with crude preparations, was confirmed using different techniques such as stepwise ammonium sulfate precipitation, hydroxyapatite chromatography and gel filtration on Sephadex G-200. The latter method permitted the separation of five glucocorticoid binding macromolecules respectively named binders S1, S2, S3, S4 and S5, according to their decreasing apparent molecular size upon gel filtration. Apparent molecular weight, binding affinity, capacity and specificity of these five moieties were examined. In addition, S-aryl-transferase activity using glutathione as co-substrate was studied and found to coincide mostly with the fractions containing binder S4, which might represent an avian liver ligandin.

Reversibility of the phosphate transfer between ATP and phosphoproteins catalysed by a cyclic nucleotide independent (G type) casein kinase.

Purified casein kinase G was found able to catalyse the synthesis of [gamma-32P]ATP in the presence of ADP, phosphocasein (previously 32P-labeled by the forward kinase reaction) and magnesium. Apparent Km values of approx. 0.5 mM for phosphocasein and 7.5 mM for ADP were calculated, these values indicating low affinities for the substrates as compared to those exhibited for casein and ATP in the forward reaction. The reverse casein kinase G activity appeared to prefer ADP and GDP as phosphate acceptors. Whereas the casein kinase G reverse reaction could be supported by casein, phosvitin and histone previously phosphorylated by the enzyme, the same proteins could not serve as a phosphate source when previously phosphorylated by the cAMP-dependent protein kinase. Forward and reverse casein kinase G reactions exhibited different optimal pH values (8.5 and 7.2, respectively) and a different sensitivity to Mg2+. Spermine, which activated the kinase activity, blocked the reverse reaction at millimolar concentrations. Although the biological significance of the casein kinase G reverse activity remains to be assessed in intact cell, the process may be useful as a tool in the characterization of phosphorylatable sites in phosphoproteins.

Catalytic and molecular properties of a highly purified G type casein kinase from bovine lung tissue.

A preparation procedure has been worked out to obtain a highly purified G type (using GTP as well as ATP) casein kinase from large quantities of bovine lung tissue. It included ion-exchange (DEAE and phosphocellulose) and affinity (casein and ATP-Sepharose) chromatography combined with a flocculation step, and yielded an apparently homogeneous preparation with a 16% yield and a purification factor of more than 1400. The purified lung casein kinase used GTP (Km 16 microM) almost as well as ATP (Km 6.7 microM) and exhibited the major catalytic properties of the casein kinase G previously described in bovine adrenal cortex (Cochet, C., Job, D., Pirollet, F. and Chambaz, E.M. (1981) Biochim. Biophys. Acta 658, 191-201). Mg2+ (30-50 mM) and spermine (2 mM) were potent activators of lung casein kinase G activity, whereas the enzyme was inhibited by heparin and quercetin. The purified enzyme underwent self-phosphorylation in the presence of ATP or GTP, serine being the only target amino acid under these conditions, whereas both serine and threonine were phosphorylated by the enzyme in casein. Lung casein kinase G exhibited an apparent molecular weight between 140 000-160 000 upon gel filtration and appeared formed by the association of two different subunits upon SDS-polyacrylamide gel electrophoresis. The two subunits of Mr 38 000 (alpha) and 27 000 (beta) exhibited a 2:1 ratio upon quantitative scanning, suggesting an alpha 3 beta 2 combination in the oligomeric native enzyme structure. Peptide mapping of the two isolated subunits following 125I-labeling and papain digestion did not disclose any common fragment. The casein kinase catalytic activity was found associated with the alpha (38 kDa) enzyme subunit after recovery from gel electrophoresis in the presence of SDS, whereas the 27 kDa (beta) subunit was the major target of the enzyme self-phosphorylation reaction. alpha and beta subunits appeared strongly associated in the oligomeric enzyme and the possible role of the beta subunit in the casein kinase G activity remains to be examined. The purified casein kinase G, which can be obtained by the present procedure, should facilitate the study of the biological significance of this phosphorylation system in the intact cell.

Cyclic nucleotide independent casein kinase (G type) in bovine adrenal cortex: purification and properties of two molecular forms.

Two soluble cyclic nucleotide independent protein kinase (ATP: protein-phosphotransferase, EC 2.7.1.37) activities have been purified from bovine adrenal cortex cytosol. Both purified enzymes exhibit the best affinity for acidic substrates such as casein and can use GTP as well as ATP as phosphoryl donor. They can thus be classified as casein kinase of the G type as previously proposed (Cochet C. et al., (1980) Endocrinology 106, 750-757). Whereas the two moieties could be separated using their different affinities toward a phosphocellulose resin, both purified enzymes appeared indistinguishable on the basis of several molecular and catalytic properties. Both G type casein kinase moieties have an identical sedimentation behavior (5.5 S in the presence of 0.5 M NaCl), yield similar patterns upon electrophoresis under denaturing conditions with three major protein components (42 000, 38 000 and 27 000), and show an ability to undergo self-phosphorylation mostly on the 27 000 component. Both enzymes have the same protein and nucleotide (ATP and GTP) substrate specificity, show similar increases in activity in the presence of polyamines and Mg2+ (optimum at 50 mM) and similar inhibition by NaCl above 0.2 M. The only difference between the two forms of casein kinase (i.e., affinity for phosphocellulose) could not be explained by a different degree of self-phosphorylation or by a limited proteolytic process during handling and purification. These results suggest that the two active moieties may represent isoenzymatic forms of the G type casein kinase activity in bovine adrenal cortex cytosol.

The tight association of protein kinase CK2 with plasma membranes is mediated by a specific domain of its regulatory beta-subunit.

Previous immunocytochemical studies have shown that protein kinase CK2 is mostly detected both in the cytoplasm and the nucleus of most cells. In the present study, CK2 was detected in highly purified plasma membrane preparations from rat liver. The protein kinase could be released from the membranes by high salt extraction (>1 M NaCl). Plasma membranes prepared from SF9 insect cells expressing the alpha- and beta-subunits of CK2 also contained a significant amount of oligomeric CK2. Furthermore, it was demonstrated in this cell system as well as in rat liver plasma membranes, that the beta-subunit of the kinase is the targeting subunit which mediates the tight association of the enzyme to plasma membrane components. Binding studies using membranes and recombinant proteins corresponding to different regions of the beta-subunit suggest that a functional domain previously shown to be involved in the binding of polyamines may also participate to the binding of CK2 to membranes. Modification of membranes by trypsin and phospholipases indicated that the binding process may require both membrane protein(s) and phospholipids. Interestingly, it was observed that the amount of membrane-bound CK2 in liver of embryos and new born rats increases dramatically after birth and persists during the postnatal stages of development.

Protein kinase-C in the human fetal adrenal gland.

The fetal zone (FZ) of the human fetal adrenal gland undergoes rapid growth and exhibits a high rate of steroidogenesis throughout fetal life. In addition to cAMP-dependent processes regulating steroidogenesis and possibly growth of the FZ, evidence is accumulating that cAMP-independent mechanisms are also involved. The purpose of this study was to determine if the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), a potent stimulator of protein kinase-C activity, stimulates steroidogenesis in FZ cells and to characterize protein kinase-C activity in FZ, neocortex zone, and anencephalic adrenal tissues. Adrenal glands were obtained from first and second trimester abortions and two anencephalic fetuses. The FZ was dissected from the neocortex. In some experiments, dispersed FZ cells were incubated in the presence and absence of ACTH and TPA for 3 h. TPA and ACTH stimulated steroidogenesis 2- and 5-fold, respectively. In other experiments, the separated zones and anencephalic adrenal tissues were homogenized, and the homogenates were subjected to DEAE-cellulose column chromatography. A single peak with phospholipid- and calcium-dependent activity was found. Subcellular distribution studies demonstrated greatest activity in the cytosolic fraction. The specific activity of protein kinase-C was significantly greater in FZ than neocortex zone, whether expressed per mg protein or per microgram DNA content. The activity in anencephalic tissue was low. In addition, protein kinase-C (80,000-dalton molecular size protein) was detected in adrenal tissues after electrophoresis and immunoblotting using an antibody directed against protein kinase-C. Greater amounts of protein kinase-C were detected in FZ tissue than in NC or anencephalic adrenal tissue. These results indicate that the lower activities of protein kinase-C in neocortex and anencephalic adrenal tissues were due to low amounts of enzyme rather than inactive enzyme. In summary, TPA-stimulated steroidogenesis in fetal zone cells and fetal zone cells contained greater activity and a greater amount of protein kinase-C than neocortex cells. Minimal activity and enzyme protein were found in anencephalic tissues. These results suggest that cAMP-independent mechanisms may play a role in fetal adrenal steroidogenesis.

Selective inhibition of a cyclic nucleotide independent protein kinase (G type casein kinase) by quercetin and related polyphenols.

The effect of quercetin and a number of structurally related phenolic compounds upon the activity of three different purified protein kinases was examined. Whereas the catalytic subunit of a cyclic AMP-dependent protein kinase and an A type (using only ATP) cyclic nucleotide-independent casein kinase (CKA) were not affected, a G type (using GTP as well as ATP) casein kinase (CKG) was selectively inhibited by several bioflavonoid structures. Kinetic studies showed that quercetin behaved as a competitive inhibitor toward the nucleotidic substrate and exhibited a high affinity for the ATP (Ki = 0.75 microM) and GTP (Ki = 0.22 microM) site of the enzyme. Considering the CKG inhibitory potency of a series of flavonoid, cinnamic acid and coumarin derivatives, it is suggested that the biological activity lays upon a common structural feature involving a phenolic ring bearing a side chain with conjugated double bonds and an oxygenated function, as found in the coumaroyl residue. These observations suggest that quercetin and related compounds may lead to a shift in intracellular protein phosphorylations by selectively inhibiting a particular type of protein kinase activity (CKG). It remains to be established whether this process may contribute to the mechanism of action of flavonoids upon cellular metabolism, particularly in the case of malignant cells.

Polyamine-mediated protein phosphorylations: a possible target for intracellular polyamine action.

Polyamines are well-known ubiquitous components of living cells. Although these polycations have been implicated in the regulation of major cellular functions such as DNA, RNA and protein synthesis occurring during cellular proliferation and/or differentiation processes, their mechanism of action at the molecular level has remained obscure. On the other hand, protein phosphorylation has emerged as a regulatory process of prime importance in cellular regulation. Data have recently been presented suggesting that polyamines may express at least part of their biological action through an effect upon selective protein phosphorylation systems. Two types of polyamine-sensitive protein kinases have been characterized in the last few years. The best known in molecular terms is the widespread casein kinase G (also termed casein kinase II), which represents a multifunctional protein kinase, at present classified as a messenger-independent activity. The other is a polyamine-dependent nuclear ornithine decarboxylase kinase characterized in Physarum polycephalum and several mammalian tissues. Both protein kinases are activated by polyamines in vitro at concentrations compatible with a physiological role, by a mechanism which most likely also involves an effect through the protein substrate conformation. Preliminary evidence suggests that both kinases may be implicated in the regulation of DNA-dependent RNA polymerase activities, although several other potential substrates have been suggested for casein kinase G. Another suggestion is that these kinases may also participate in the post-translational regulation of ornithine decarboxylase, the rate-limiting step in the polyamine biosynthetic pathway. A novel class of protein kinase activities may thus be defined as polyamine-mediated phosphorylation systems for which polyamines may function as intracellular messenger. Although their biological significance remains to be fully established, especially with regard to the definition of their specific intracellular target(s) and subsequent biological functions, these systems will be interesting to consider in future studies aimed at understanding the role of polyamines in cell regulation.

Effects of transforming growth factor beta on ovine adrenocortical cells.

Transforming growth factor beta (TGF beta) is a potent regulator of steroidogenic cell function. However, the mechanisms of the effects are not well understood. We studied the actions of TGF beta on primary cultures of ovine adrenocortical (OAC) cells. OAC cells had high affinity receptors for TGF beta (KD congruent to 7.6 +/- 1.5 X 16(-11) M). In addition, TGF beta inhibited the following markers of adrenocortical function: (1) ACTH, cholera toxin and forskolin acute stimulation of cAMP and steroid production; (2) the acute 8-bromo-cAMP stimulation of corticosteroid and pregnenolone production; and (3) the activity and amount of P-450 17 alpha-hydroxylase protein as well as activities of 11 beta- and 21-hydroxylases. The inhibitory effects of TGF beta on ACTH-induced cAMP and steroid production were time (half inhibition at 6 and 3 h respectively) and dose dependent (ID50 congruent to 10(-12) M). From these data we concluded that TGF beta acted rapidly on sites of OAC cell acute responses to stimulation by ACTH before and after the production of cAMP. Pregnenolone production in these cells was not inhibited by TGF beta when steroid production was stimulated on the addition of the readily permeable cholesterol derivative, 22 R-hydroxycholesterol. Thus, the rapid effect on OAC cells was manifest by TGF beta action on the utilization of cellular pools of cholesterol for the acute stimulation of steroid formation and not by direct action on the cholesterol side-chain cleavage enzyme. In addition, cells stimulated with ACTH in the absence or presence of lipoproteins (for up to 36 h) were susceptible to the inhibitory action of TGF beta. Taken together, these data amplify the pleiotropic actions of TGF beta on adrenocortical cell function and demonstrate that one acute action of TGF beta is on the utilization of endogenous supplies of cholesterol for steroid production.

Homocysteine, hypothyroidism, and effect of thyroid hormone replacement.

Elevation of total plasma concentration of homocysteine (t-Hcy) is an important and independent risk factor for cardiovascular disease. Hypothyroidism is possibly also associated with an increased risk for coronary artery disease, which may be related to atherogenic changes in lipid profile. Because hypothyroidism decreases hepatic levels of enzymes involved in the remethylation pathway of homocysteine, we prospectively evaluated fasting and postload t-Hcy in patients before and after recovery of euthyroidism. Fasting and postload t-Hcy levels were higher in 40 patients with peripheral hypothyroidism (14 with autoimmune thyroiditis and 26 treated for thyroid cancer) in comparison with those of 26 controls (13.0 +/- 7.5 vs. 8.5 +/- 2.6 micromol/L, p < .01, respectively, and 49.9 +/- 37.3 vs. 29.6 +/- 8.4 micromol/L p < .001, respectively). On univariate analysis, fasting Hcy was positively related to thyrotropin (TSH) and inversely related to folates. Multivariate analysis confirmed TSH as the strongest predictor of t-Hcy independent of age, folate, vitamin B12, and creatinine. Thyroid hormone replacement significantly decreased fasting but not postload t-Hcy. We conclude that t-Hcy is elevated in hypothyroidism. The association of hyperhomocysteinemia and lipid abnormalities occurring in hypothyroidism may represent a dynamic atherogenic state. Thyroid hormone failed to completely normalize t-Hcy. Potential benefit of treatment with folic acid in combination with thyroid hormone replacement has to be tested given that hypothyroid patients were found to have lower levels of folate.

Effects of carbon dioxide on the fate of Listeria monocytogenes, of aerobic bacteria and on the development of spoilage in minimally processed fresh endive.

Minimally processed fresh broad-leaved endive (Cichorium endivia L.) were stored at 3 and 10 degrees C in modified atmospheres containing air, 10% CO2/10% O2, 30% CO2/10% O2, and 50% CO2/10% O2. The effects of these modified atmospheres on the fate of both aerobic bacteria and three strains of Listeria monocytogenes, was investigated. Increases in CO2 concentrations significantly reduced the growth of the aerobic microflora. The best preservation of the visual quality occurred on endive leaves stored in 10% CO2/10% O2, whereas leaves stored in 30% CO2/10% O2 and 50% CO2/10% O2, and to a lesser extent in air, showed extensive spoilage after storage. Listeria monocytogenes was slightly affected at 3 degrees C by the modified atmospheres, as compared to air. At 10 degrees C, results varied between replicate experiments, but L. monocytogenes generally grew better as the CO2 concentration was increased. The three test strains behaved in a similar way. In conclusion, among the modified atmospheres tested, a modified atmosphere containing 10% CO2/10% O2 resulted in improved visual quality of minimally processed fresh endive, without a marked effect on the growth of the aerobic microflora or of L. monocytogenes.

[Cyclic AMP-dependent and -independent protein kinases in the adrenal cortex]. / Protéines kinases AMP-cyclique dépendantes et indépendantes dans le cortex surrénal.

The various protein kinase activities characterized in adrenal cortex from different mammalian species do not differ in their molecular and catalytic properties from similar types of enzymes described in other tissues. The cyclic AMP-dependent system has been the most thoroughly investigated in adrenocortical tissue and is generally accepted as an obligatory intermediate in the intracellular response to ACTH action. In addition, adrenocortical cells contain two types of messenger independent protein kinases, one of which being selectively activated by polyamines in vitro and possibly related to the control of adrenocortical growth processes. The definition of intracellular phosphorylatable targets related to specific adrenocortical functions might have been obscured by the pleiotypic effects displayed by ACTH, the major effector of these cell activities. Cholesterol esterase appears one of these targets and 11 beta-hydroxylase cytochrome P-450 has been found activated in vitro by a cAMP dependent process. Future research should provide a better understanding of the role of protein phosphorylation-dephosphorylation processes as a regulatory device in adrenocortical cell functions. This includes possible complex modulation by multisite phosphorylations of a given target protein, involving several different protein kinase systems under the dependence of separate intracellular effectors.