[Social reimbursement-the Spanish-German ENT Society's (SDGHNO) Latin America project]. / Soziale Rückvergütung ­ das Projekt "Lateinamerika" der Spanisch-Deutschen HNO-Gesellschaft (SDGHNO).

Nowadays, social projects are usually oriented in such a way that after a given period of time, they can either support themselves independently or even allow a pecuniary reimbursement. In the latter case, experts speak of a profit-oriented reimbursement. On the other hand, there is so-called social reimbursement, which in contrast to the abovementioned form is not profit oriented, but, for example, considers its task fulfilled by the fact of successful knowledge transfer. The Spanish-German Society for ENT Medicine and Head and Neck Surgery (SDGHNO) launched the Latin America project in 2001 under the patronage of the then President Prof. Dr. Wolfgang Draf (Fulda). The goal of the SDGHNO was and is to create a professional as well as cultural platform for Spanish- and German-speaking ENT doctors. This platform can and should be used for professional purposes, e.g., for knowledge transfer. Since the beginning of its existence, the Latin America project has thus brought numerous scientific events into being and created specific contacts which have lasted until today or have even been continued and further developed. Particularly successful examples are Chile, Colombia, and Peru. This is a vivid example of social reimbursement, because the participating German-speaking members/speakers carried out their tasks on an entirely voluntary basis. Thus, the SDGHNO did not bear any travel, catering, or accommodation costs. The activities of the SDGHNO within the framework of the Latin America project are explained.

IQGAP1 is necessary for pulmonary vascular barrier protection in murine acute lung injury and pneumonia.

We recently reported that integrin α(v)ß(3) is necessary for vascular barrier protection in mouse models of acute lung injury and peritonitis. Here, we used mass spectrometric sequencing of integrin complexes to isolate the novel ß(3)-integrin binding partner IQGAP1. Like integrin ß(3), IQGAP1 localized to the endothelial cell-cell junction after sphingosine-1-phosphate (S1P) treatment, and IQGAP1 knockdown prevented cortical actin formation and barrier enhancement in response to S1P. Furthermore, knockdown of IQGAP1 prevented localization of integrin α(v)ß(3) to the cell-cell junction. Similar to ß(3)-null animals, IQGAP1-null mice had increased pulmonary vascular leak compared with wild-type controls 3 days after intratracheal LPS. In an Escherichia coli pneumonia model, IQGAP1 knockout mice had increased lung weights, lung water, and lung extravascular plasma equivalents of (125)I-labeled albumin compared with wild-type controls. Taken together, these experiments indicate that IQGAP1 is necessary for S1P-mediated vascular barrier protection during acute lung injury and is required for junctional localization of the barrier-protective integrin α(v)ß(3).

"Full moon" endoscopic sign in intraventricular neurocysticercosis.

INTRODUCTION: Despite improvements in sanitation, diagnosis and treatment, neurocysticercosis is still a public health problem in many countries. In symptomatic patients, there is a broad spectrum of clinical manifestations. When cysticerci are lodged in the ventricles or the subarachnoid space, the flow of cerebrospinal fluid can be obstructed and lead to hydrocephalus and intracranial hypertension. The endoscopic view may be useful as a diagnostic tool. PATIENTS: This report clearly shows a common endoscopic pattern in 4 selected patients with ventricular cysticercosis (2 third ventricle/2 lateral ventricle). The endoscopic view of the cysts in the ventricles resembles a "full moon". This analogy helped to identify the features of cysticerci with intact walls and the vesicular stage, malleable due to its cystic content and having an irregular surface, as evidence of the microscopic structure of the cyst wall in a cysticercus. CONCLUSIONS: This finding is not seen in other intraventricular cysts or tumors that can actually be considered as an additional diagnostic criterion among the definitive findings to establish the diagnosis of cysticercosis, since it involves direct endoscopic visualization of a cysticercus under histopathological demonstration. Additionally, the endoscopic approach can be used as primary treatment for these cases, following the minimally invasive approach principle.

Carbon source-dependent phosphorylation of hexokinase PII and its role in the glucose-signaling response in yeast.

The HXK2 gene is required for a variety of regulatory effects leading to an adaptation for fermentative metabolism in Saccharomyces cerevisiae. However, the molecular basis of the specific role of Hxk2p in these effects is still unclear. One important feature in order to understand the physiological function of hexokinase PH is that it is a phosphoprotein, since protein phosphorylation is essential in most metabolic signal transductions in eukaryotic cells. Here we show that Hxk2p exists in vivo in a dimeric-monomeric equilibrium which is affected by phosphorylation. Only the monomeric form appears phosphorylated, whereas the dimer does not. The reversible phosphorylation of Hxk2p is carbon source dependent, being more extensive on poor carbon sources such as galactose, raffinose, and ethanol. In vivo dephosphorylation of Hxk2p is promoted after addition of glucose. This effect is absent in glucose repression mutants cat80/grr1, hex2/reg1, and cid1/glc7. Treatment of a glucose crude extract from cid1-226 (glc7-T152K) mutant cells with lambda-phosphatase drastically reduces the presence of phosphoprotein, suggesting that CID1/GLC7 phosphatase together with its regulatory HEX2/REG1 subunit are involved in the dephosphorylation of the Hxk2p monomer. An HXK2 mutation encoding a serine-to-alanine change at position 15 [HXK2 (S15A)] was to clarify the in vivo function of the phosphorylation of hexokinase PII. In this mutant, where the Hxk2 protein is unable to undergo phosphorylation, the cells could not provide glucose repression of invertase. Glucose induction of HXT gene expression is also affected in cells expressing the mutated enzyme. Although we cannot rule out a defect in the metabolic state of the cell as the origin of these phenomena, our results suggest that the phosphorylation of hexokinase is essential in vivo for glucose signal transduction.

[A randomized single-blind trial of the effects of vitamins C and E in familial hypercholesterolemia]. / Ensayo aleatorizado ciego-sencillo sobre los efectos de las vitaminas C y E en la hipercolesterolemia familiar.

INTRODUCTION: The aim of this study was to evaluate the effect of antioxidant vitamin C and E administration on dyslipidemia, plasma fatty acid composition, and biochemical inflammatory markers in children with heterozygous familial hypercholesterolemia (FH). PATIENTS: Forty girls and boys with heterozygous FH, aged between 2 and 18 years, and with plasma low-density lipoprotein (LDL)-cholesterol levels higher than 160 mg/dl were studied. METHODS: We performed an open longitudinal randomized trial over a 1-year period. All children followed a dietary intervention according to the National Cholesterol Education Program (NCEP)-1 guidelines and were randomized into two groups. One group (n = 21) received therapy with vitamin C (500 mg twice a day) and vitamin E (400 IU per day). A second group (n = 19) did not receive vitamin therapy. RESULTS: In patients receiving antioxidant vitamins, plasma linoleic acid levels (18:2 omega-6) significantly increased and the essential fatty acid deficiency ratio significantly decreased (Mead/arachidonic acid: 20:4 omega-6/20:3 omega-9). No significant differences were observed in plasma lipid profile, adhesion molecules, or reactive C protein. CONCLUSIONS: Antioxidant vitamin therapy in children with heterozygous FH modifies the plasma fatty acid profile. These modifications are independent of the degree of dyslipidemia and may represent an indicator of reduced cardiovascular risk.

Prognostic value of gasometric parameters of carbon dioxide in resuscitation of septic patients. A bibliography review.

The anaerobic metabolism is the cornerstone in physiopathology of septic shock. Nowadays we have both the central or mixed venous oxygen saturation and lactate levels to monitoring the metabolism in septic patients. Some studies have shown that normalization of systemic hemodynamic and oxygen metabolism variables not prevent progression to multiorgan damage and death. Recently has been proposed the venous-to-arterial carbon dioxide difference (ΔpvaCO2) as an alternative marker of tissue hypoperfusion, like Cardiac Index. High ΔpvaCO2 predicts adverse outcomes. Also has been proposed both, the ratio between the ΔpvaCO2 and arterial-to-venous oxygen content difference (ΔCavO2): ΔpvaCO2/ΔCavO2; and, the ratio between venous-to-arterial carbon dioxide difference (ΔCvaCO2) and ΔCavO2: ΔCvaCO2/ΔCavO2, as markers of anaerobic metabolism. Both of high ratios are related to high levels of lactate and worse prognosis. Therefore in patients with sepsis the combination of markers of resuscitation could be important to improve the outcomes.

Oncogenic KIT mutations in different exons lead to specific changes in melanocyte phospho-proteome.

UNLABELLED: Mutations in the proto-oncogene c-KIT (KIT) are found in several cancers, and the site of these mutations differs markedly between cancer types. We used site directed mutagenesis to induce KIT(559), KIT(642) and KIT(816) mutations in primary human melanocytes (PHM) and we investigated the impact of each mutation on KIT function. We studied canonical KIT-signaling pathways by immunoblotting, and we used stable isotope labeling by amino acids in cell culture (SILAC) and kinase prediction models to identify kinases differently activated in respective mutants. We validated our results with the analysis of phosphorylation levels of selected substrates for each kinase. We concluded that CK1 ε and δ are more active in cell clones harboring KIT(559) and KIT(642) mutations, whereas PAK4 is more active in clones with KIT(816) mutation. Our findings might help to develop further therapeutic options for tumors with specific KIT mutations in different domains. BIOLOGICAL SIGNIFICANCE: Different types of cancers harbor mutations in the oncogene KIT. The use of small molecules inhibitors directly targeting KIT had a limited success in the treatment of patients with KIT mutant cancers. Our study describes specific phospho-proteome changes due to different KIT mutations, and provides targets of further therapeutic options.

Engineering baker's yeast: room for improvement.

Bread making is one of the oldest food-manufacturing processes. However, it is only in the past few years that recombinant-DNA technology has led to dramatic changes in formulation, ingredients or processing conditions. New strains of baker's yeast that produce CO2 more rapidly, are more resistant to stress or produce proteins or metabolites that can modify bread flavour, dough rheology or shelf-life are now emerging.

Hexokinase PII has a double cytosolic-nuclear localisation in Saccharomyces cerevisiae.

We describe here that the HXK2 gene product, isoenzyme PII of hexokinase, is located in both the nucleus and the cytoplasm of Saccharomyces cerevisiae cells. This conclusion is supported by assays of hexokinase-specific activity in isolated nuclei from wild-type and hxk1lhxk2 double mutant strains, by immunoblot experiments using anti-Hxk2 antibodies and by observation of the fluorescence distribution of a Hxk2-GFP fusion protein in cells transformed with the HXK2::gfp gene.

p38 MAPK mediates the regulation of alpha1(I) procollagen mRNA levels by TNF-alpha and TGF-beta in a cell line of rat hepatic stellate cells(1).

The role of members of the mitogen-activated protein kinase (MAPK) family on tumor necrosis factor alpha (TNF-alpha)-mediated down-regulation of col1a1 gene was studied. TNF-alpha increased extracellular-regulated kinase and Jun-N-terminal kinase phosphorylation, but these effects were not related to its inhibitory effect on alpha1(I) procollagen (col1a1) mRNA levels. Phosphorylation of p38 MAPK was decreased in response to TNF-alpha, and the specific p38 MAPK inhibitor SB203580 mimicked the effect of TNF-alpha on col1a1 mRNA levels. Transforming growth factor beta (TGF-beta) increased p38 MAPK phosphorylation and SB203580 prevented the induction of col1a1 mRNA levels by TGF-beta. These results suggest that p38 MAPK plays an important role in regulating the expression of col1a1 in hepatic stellate cells in response to cytokines.

A randomized trial of prophylactic doxycycline for curettage in incomplete abortion.

OBJECTIVE: To determine whether prophylactic doxycycline at suction curettage for incomplete abortion decreases the rate of postoperative pelvic infection. METHODS: We randomized 240 patients to receive intravenous doxycycline or placebo at curettage. Cervical specimens for gonorrhea and chlamydia were obtained preoperatively. Two weeks post-procedure, we evaluated all patients for infectious morbidity and repeated gonorrhea and chlamydia cultures. Statistical analysis used Mann-Whitney U test, McNemar test, or Fisher exact test, as appropriate. RESULTS: There were no statistically significant differences in age, parity, gestational age, history of sexually transmitted disease, pelvic inflammatory disease, or multiple sex partners between the doxycycline and placebo groups. Preoperative gonorrhea or chlamydia isolates were positive in five (4.2%) and six (5%) of 120 doxycycline patients and four (3.3%) and eight (6.6%) of 120 controls (not significant). All preoperative gonorrhea isolates remained positive postoperatively. Seven (5.8%) controls had positive postoperative chlamydia isolates, as did one (0.8%) in the doxycycline group (P = .06). We diagnosed eight (6.6%) of 120 doxycycline patients and seven (5.8%) of 120 controls with infectious morbidity (not significant). CONCLUSION: In our population of patients with incomplete abortion, the prevalence of gonorrhea and chlamydia was low, and prophylactic doxycycline did not decrease the rate of postoperative febrile morbidity.

Nucleotide sequence of a putative peroxisomal protein from the yeast Lipomyces kononenkoae.

A cDNA corresponding to a putative peroxisomal protein from the yeast Lipomyces kononenkoae has been isolated. It contains an ORF of 498 bp that predicts a polypeptide of 166 amino acids with an apparent molecular mass of 17,365 Da. The protein shares a high degree of homology with proteins associated to the peroxisomal membrane of Candida boidinii.

A new NPY-antagonist strongly stimulates apoptosis and lipolysis on white adipocytes in an obesity model.

Neuropeptide Y (NPY) is a 36 amino acid peptide released in central and peripheral mammalian neurons, which appears to contribute to adiposity regulation by increasing food intake, thus promoting weight gain on animals. Nevertheless, little is known about NPY direct actions on white adipocytes. This trial, which was designed to test the possible effects of a new NPY antagonist, S.A.0204, on white adipose tissue, revealed that the administration of this novel molecule strongly ex vivo stimulates apoptosis and lipolysis in animals fed on a high-fat diet.

Determination of 5-dodecylsalicylaldoxime and 5-nonylsalicylaldoxime in commercial extractants by high-performance liquid chromatography with photometric detection.

A liquid chromatographic method with photometric detection was developed and validated for the determination of 5-dodecylsalicylaldoxime in the industrial extractants LIX 622 and LIX 860 and 5-nonylsalicylaldoxime in LIX 622N, Acorga PT5050 and Acorga P5100. The limit of detection was 2.18 microg/ml for 5-dodecylsalicylaldoxime and 2.06 microg/ml for 5-nonylsalicylaldoxime. Good linear relations between the areas and the concentration of the standard injected were found in the range of 5-100 microg/ml and reproducibility studies yielded relative standard deviations lower than 6.12% in all the cases.

Capillary zone electrophoresis applied to the determination of the angiotensin-converting enzyme inhibitor cilazapril and its active metabolite in pharmaceuticals and urine.

A capillary zone electrophoresis method has been developed for the quantitation of antihypertensive drug cilazapril and its active metabolite cilazaprilat in pharmaceuticals and urine. The separation of the compounds was performed in a fused-silica capillary filled with the running electrolyte, which consisted of a 60 mM borate buffer solution at pH 9.5. Under the optimized experimental conditions, the separation took less than 5 min. The analysis of urine samples required a previous solid-phase extraction step using C8 cartridges. The method was successfully applied to the determination of the drug and its metabolite in urine samples obtained from three hypertensive patients (detection limits of 115 ng ml(-1) for cilazaprilat and 125 ng ml(-1) for cilazapril) and to pharmaceutical dosage forms. The method was validated in terms of reproducibility, linearity and accuracy.

Stable high-copy-number integration of Aspergillus oryzae alpha-AMYLASE cDNA in an industrial baker's yeast strain.

The Aspergillus oryzae alpha-amylase cDNA was placed under the control of the Saccharomyces cerevisiae actin promoter (pACT1) and introduced into the ribosomal DNA locus of an industrial baker's yeast strain. To obtain a strain eligible for commercial use, we constructed an integrative cassette lacking bacterial DNA sequences but containing the alpha-amylase cDNA and ribosomal DNA sequences to target the integration to this locus. High-copy-number integrants were obtained including a defective TRP1d promoter in the integrative cassette. We selected one transformant, Rib-AMY (CECT10872), in which the multi-integrated sequences were stable even after 200 generations of growth in nonselective medium. This transformant also expressed and secreted high levels of alpha-amylase. Bread made with this strain had a higher volume, lower density, and softer crumbs than bread made with a control strain. The Rib-AMY transformant also was useful in retarding bread firming. This new strain fulfills all the requirements for commercial utilization and should reduce or eliminate the requirement for addition of exogenous alpha-amylase to the flour, reducing allergenic work-related symptoms due to this enzyme.

The value of exercise electrocardiography testing in the identification of coronary restenosis: a probability analysis.

We studied by means of probability analysis the role of exercise ECG in identifying coronary restenosis. A total of 213 patients were independently evaluated by clinical history, conventional assessment of the exercise ECG ('yes or no' statement), D score (a discriminant function derived from exercise ECG), and coronariography, 5.4 +/- 2.8 months after successful coronary angioplasty. The initial probability of restenosis (30%), that is, the prevalence of the condition, was radically changed by the result of clinical history (77% for patients with angina vs. 17% for those without angina). By contrast, ECG binary assessment, due to its low accuracy (70% vs 82% of clinical history, P < 0.005), was unable to significantly change the established probabilities after symptomatic evaluation. Finally, D score, which greatly enhanced specificity (92% vs. 76% of bivariate assessment, P < 0.0001), proved to be useful in changing the probability (from 32% to 76% or to 25%) of patients (n = 34) with a discordant result (no angina/positive exercise ECG). When this stepwise approach was tested in 46 new patients, predicted and observed probabilities were actually very similar. We conclude that exercise ECG has a very limited role in identifying coronary restenosis if positive responses are not adjusted with a weighted score which takes into account other exercise derived factors.

Isolation, purification, and characterization of a cold-active lipase from Aspergillus nidulans.

Aspergillus nidulans WG312 strain secreted lipase activity when cultured in liquid media with olive oil as carbon source. Highest lipase productivity was found when the mycelium was grown at 30 degrees C in a rich medium. The new enzyme was purified to homogeneity from the extracellular culture of A. nidulans by phenyl-Sepharose chromatography and affinity binding on linolenic acid-agarose. The lipase was monomeric with an apparent M(r) of 29 kDa and a pI of 4.85 and showed no glycosylation. Kinetic of enzyme activity versus substrate concentration showed a typical lipase behavior, with K(M) and K(cat) values of 0.28 mM and 494 s(-)(1) and 0.30 mM and 320 s(-)(1) for the isotropic solution and for the turbid emulsion, respectively. All glycerides assayed were hydrolyzed efficiently by the enzyme, but this showed preference toward esters of short- and middle-chain fatty acids. The optimum temperature and pH for the lipolytic activity were 40 degrees C and 6.5, with high activity in the range 0-20 degrees C and reduced thermal stability.

Expression of LIP1 and LIP2 genes from Geotrichum species in Baker's yeast strains and their application to the bread-making process.

Lipolytic baker's yeast strains able to produce extracellular active lipase have been constructed by transformation with plasmids containing the LIP1 and LIP2 genes from Geotrichum sp. under the control of the Saccharomyces cerevisiae actin promoter (pACT1). Lipase productivity differed between both constructs, YEpACT-LIP1-t and YEpACT-LIP2-t, being higher for the strain bearing the LIP2 gene in all culture media tested. This result appeared not to be the consequence of a defect in the transcription of the LIP1 gene as revealed by Northern blot analysis. Replacing the signal sequence of LIP1 by that of LIP2 in the YEpACT-LIP1-t plasmid enhanced significantly the secretion of lipase 1, but the levels of lipase activity were still lower than those found for the YEpACT-LIP2-t transformant. Recombinant lipase 2 protein produced by baker's yeast exhibited biochemical properties similar to those of the natural enzyme. Fermented dough prepared with YEpACT-LIP2-t-carrying cells rendered a bread with a higher loaf volume and a more uniform crumb structure than that prepared with control yeast. These effects were stronger by the addition in the bread dough formulas of a preferment enriched in recombinant lipase 2.

Neutrophil activation in preeclampsia. Are defensins and lactoferrin elevated in preeclamptic patients?

OBJECTIVE: To determine if human defensins and lactoferrin, both markers of neutrophil activation, are elevated in preeclamptic plasma. STUDY DESIGN: Blood samples were obtained from 18 preeclamptic and 29 normal pregnant women in the third trimester. Demographic and clinical data were obtained from the medical record. No patient had evidence of labor and/or infection when blood was drawn. Preeclamptic patients were defined by elevated blood pressure, 140/90 mm Hg, proteinuria of > 300 mg in a 24-hour collection and hyperuricemia. Human defensins and lactoferrin were measured by enzyme immunoassay of plasma samples diluted 1:100 and 1:10, respectively. Standard curve values ranged from 0.25 to 16 ng/mL. Data for human defensins and lactoferrin are presented as median values +/-SE. Statistical analysis included Student's t test for comparison of clinical data, Mann-Whitney U test for comparison of absolute values between groups and Fisher's exact test, when appropriate. RESULTS: There was no difference in age or estimated gestational age between the two groups. There were more nulliparous patients in the preeclamptic group. Human defensin levels were significantly elevated (P = .005) in plasma of preeclamptic patients (25.1 ng/mL +/- 16.2) as compared to normal controls (9.0 ng/mL +/- 8.9). Nine of 18 (50%) preeclamptic patients and 2 of 29 (7%) normal controls had defensin levels above the low point on the standard curve (P = .001). There was no difference in lactoferrin levels between the two groups. CONCLUSION: Our results suggest that preeclampsia is associated with neutrophil activation. The biologic effect of elevated human defensins in preeclamptic plasma remains to be determined.