[Subacute or initially chronic osteomyelitis in children. A series of 17 cases].
17 cases of subacute or at once chronic osteomyelitis are shown. At this occasion, an analysis of epidemiological, topographical, clinical and bacteriological data is given. An immunological study has been made in 5 cases: diagnosis is difficult at all stages. A reflexion is proposed concerning the treatment to be performed.


Decreased incidence of subglottic stenosis using an "appropriate-sized" endotracheal tube in neonates.
In a previous study of factors influencing the development of acquired subglottic stenosis (SS) we proposed choosing an endotracheal tube (ETT) size such that the ratio of the external diameter divided by the gestational age of the infant in weeks was less than 0.1. We have prospectively followed 44 patients intubated longer than 25 days. One of 36 patients intubated according to the above protocol developed significant SS compared to 3 of 8 patients intubated with an ETT larger than suggested. In a previous study, 9 of 49 patients intubated longer than 25 days developed SS. The incidence of SS in patients intubated with "appropriate-sized" ETT was less than either other group of patients (P less than 0.005 and P less than 0.05, respectively). This study confirms the suggestion that choosing ETT size by the above protocol will lessen, although by itself not eliminate, the incidence of acquired SS.


Bleeding in outpatients treated with warfarin: relation to the prothrombin time and important remediable lesions.
To determine the relation of bleeding to prothrombin times and important remediable lesions in outpatients treated with warfarin. An inception cohort of 565 patients starting outpatient therapy with warfarin on discharge from a university hospital was assembled. Detailed records of outpatient prothrombin times were obtained for 103 of 130 case subjects with major or minor bleeding and for 117 control patients without bleeding. A nested case-control design was used to evaluate the association of bleeding with temporally related prothrombin times; odds ratios were estimated using multivariate logistic regression analysis to control for known predictors of major bleeding. The relation of bleeding to important remediable lesions was determined in all 130 cases of bleeding. For each 1.0 increase in the prothrombin time-to-control ratio, the odds ratio for major bleeding during the week after a prothrombin time measurement increased 80%; the odds ratio for minor bleeding increased 50%. These odds ratios were lower during the first month of therapy and higher thereafter. Bleeding was related to important remediable lesions in 49 of 130 cases (38%), but these lesions were unknown before bleeding in only 22 cases (17%). The mean prothrombin time rose sharply at the time of bleeding in patients without important remediable lesions, but not in patients with lesions. New, previously unknown lesions (including nine malignancies) were discovered in 20 of 59 case subjects (34%) with gastrointestinal bleeding or hematuria, but in only two of 71 case subjects (3%) with other bleeding (p less than 0.001). Our results provide a valid quantitative basis for estimating the odds of bleeding in relation to the prothrombin time and the yield of diagnostic evaluation in patients with bleeding.


A Li-Fraumeni syndrome family with retained heterozygosity for a germline TP53 mutation in two tumors.
We identified a missense germline mutation (Gly245Ser) in one of the mutation hot spots of the TP53 gene in two affected members of a Li-Fraumeni syndrome family. We also analyzed their tumors, a liposarcoma and a colorectal carcinoma. Both tumors exhibited p53 protein accumulation but none of them showed loss of the wild-type allele of the TP53 gene. We reviewed all published cases of tumors in germline TP53 mutation carriers where loss of heterozygosity data were available and identified 84 tumors with loss of the wild-type allele, 57 tumors with retention of heterozygosity, and 9 tumors with loss of the allele harboring the germline mutation. Among the tumors showing p53 accumulation, we observed a significant difference in the fraction of tumors showing p53 protein accumulation between the tumors with loss of the wild-type allele and those with retention of TP53 heterozygosity. This supports the idea that the pathogenesis of tumors in germline TP53 mutation carriers does not have to be associated with loss of the wild-type TP53 allele. The product of the normal allele can potentially be inactivated by a variety of other mechanisms or, as suggested by the analysis, many of these tumors may even preserve the activity of the wild-type p53 protein.


Processing of natural fibre and method improvement for removal of endocrine-disrupting compounds.
Persistent endocrine-disrupting compounds (EDCs) in bodies of water are a concern for human health and constitute an environmental issue, even if present in trace amounts. Conventional treatment systems do not entirely remove EDCs from discharge effluent. Due to the ultra-trace level of EDCs which affect human health and pose an environmental issue, developing new approaches and techniques to remove these micropollutants from the discharged effluent is vital. This review discusses the most common methods of eliminating EDCs through preliminary, primary, secondary and tertiary treatments. The adsorption process is favoured for EDC removal, as it is an economical and straightforward option. The NABC aspects, which are the need, approach, benefits and challenges, were analysed based on existing circumstances, highlighting biochar as a green and renewable adsorbent for the removal of organic contaminants. From the environmental point of view, the effectiveness of this method, which uses natural fibre from the kenaf plant as a porous and economical biochar material with a selected lignocellulosic biomass, provides insights into the advantages of biochar-derived adsorbents. Essentially, the improvement of the natural fibre as an adsorbent is a focus, using carbonisation, activation, and the physiochemical process to enhance the adsorption ability of the material for pollutants in bodies of water. This output will complement sustainable water management approaches presented in previous studies for combating the emerging pollutant crisis via novel green and environmentally safe options.


Expression of plexin-B1 in the mouse ovary and its possible role in follicular development.
To study the expression of plexin-B1 in the mouse ovary and its possible role in follicular development. Controlled laboratory study. Research laboratory, Ha'Emek Medical Center. ICR female mice (n = 36) were studied. Plexin-B1 expression by immunohistochemistry, Western blot analysis, and reverse transcriptase-polymerase chain reaction. Microscopic morphometric and morphologic analysis for the evaluation of mouse follicular growth and E2, P, and T secretion from the developed follicles. Plexin-B1 was expressed in the mouse ovary mostly in the granulosa cells. Plexin-B1 gene expression increased significantly by 126.8% +/- 38% in the pregnant mare serum gonadotropin-treated group and increased by 98.7% +/- 15.2 and 262.2% +/- 65.4% in the hCG-treated group 6 h and 24 h after injection, respectively (ANOVA; P < .05). Comparison of the maximum diameters of the control and study groups on day 9 of the culture period revealed a significant difference between the two groups (P < .001). In the control group, the mean maximum diameter was 191 +/- 2.6 microm compared with 136 +/- 5.13 microm in the presence of neutralizing antibodies against plexin-B1. Neutralizing antibody against plexin-B1 decreases E2 and P secretion. This study shows for the first time the expression of plexin-B1 in the mouse ovary. A possible role for plexin-B1 in follicular growth and steroidogenesis, most likely via the granulosa cells, is suggested.


Comparative efficacy of propranolol and chlordiazepoxide in alcohol withdrawal.
Propranolol alone was more effective than either chlordiazepoxide or a combination of chlordiazepoxide and propranolol in alleviating the symptoms of the alcohol withdrawal syndrome.


RGC-specific ATF4 and/or CHOP deletion rescues glaucomatous neurodegeneration and visual function.
Endoplasmic reticulum (ER) stress has been linked with various acute and chronic neurodegenerative diseases. We previously found that optic nerve (ON) injury and diseases induce neuronal ER stress in retinal ganglion cells (RGCs). We further demonstrated that germline deletion of CHOP preserves the structure and function of both RGC somata and axons in mouse glaucoma models. Here we report that RGC-specific deletion of CHOP and/or its upstream regulator ATF4 synergistically promotes RGC and ON survival and preserves visual function in mouse ON crush and silicone oil-induced ocular hypertension (SOHU) glaucoma models. Consistently, topical application of the ATF4/CHOP chemical inhibitor ISRIB or RGC-specific CRISPR-mediated knockdown of the ATF4 downstream effector Gadd45a also delivers significant neuroprotection in the SOHU glaucoma model. These studies suggest that blocking the neuronal intrinsic ATF4/CHOP axis of ER stress is a promising neuroprotection strategy for neurodegeneration.


Values at stake: autonomy, responsibility, and trustworthiness in relation to genetic testing and personalized nutrition advice.
Personalized nutrition has the potential to enhance individual health control. It could be seen as a means to strengthen people's autonomy as they learn more about their personal health risks, and receive dietary advice accordingly. We examine in what sense personalized nutrition strengthens or weakens individual autonomy. The impact of personalized nutrition on autonomy is analyzed in relation to responsibility and trustworthiness. On a societal level, individualization of health promotion may be accompanied by the attribution of extended individual responsibility for one's health. This constitutes a dilemma of individualization, caused by a conflict between the right to individual freedom and societal interests. The extent to which personalized nutrition strengthens autonomy is consequently influenced by how responsibility for health is allocated to individuals. Ethically adequate allocation of responsibility should focus on prospective responsibility and be differentiated with regard to individual differences concerning the capacity of adults to take responsibility. The impact of personalized nutrition on autonomy also depends on its methodological design. Owing to the complexity of information received, personalized nutrition through genetic testing (PNTGT) is open to misinterpretation and may not facilitate informed choices and autonomy. As new technologies, personalized nutrition and PNTGT are subject to issues of trust. To strengthen autonomy, trust should be approached in terms of trustworthiness. Trustworthiness implies that an organization that develops or introduces personalized nutrition can show that it is competent to deal with both the technical and moral dimensions at stake and that its decisions are motivated by the interests and expectations of the truster.


The moderating role of trajectories of family hardiness in the relationship between trajectories of economic hardship and mental health of mothers and children.
Trajectories of perceived economic hardship are related to clinical levels of mental health issues in mothers and children from low-income families. Cross-sectionally, family hardiness has been found to have a moderating effect on the relationship between stressors and mental health severity. Recent advances in family resilience theory highlight the importance of considering trajectories of family resilience. Trajectories of family hardiness and their moderating effect on the relationship between trajectories of perceived economic hardship and symptoms of depression and anxiety in low-income mothers and children were investigated in 511 mother-child dyads in Singapore. Three trajectories of family hardiness were delineated, namely the high stable, low rapidly increasing and moderate increasing group. The trajectories of family hardiness were found to moderate the relationship between trajectories of perceived economic hardship and symptoms of mental health in low-income mothers and children. The same moderation effect was not found when perceived economic hardship and family hardiness were investigated cross-sectionally. These findings highlight the importance of considering the family's trajectory of hardiness over time when working with low-income families. In addition, given that different trajectories of family hardiness were protective for different aspects of mental health, further studies to understand these relations are necessary.


Prediction of change in protein unfolding rates upon point mutations in two state proteins.
Studies on protein unfolding rates are limited and challenging due to the complexity of unfolding mechanism and the larger dynamic range of the experimental data. Though attempts have been made to predict unfolding rates using protein sequence-structure information there is no available method for predicting the unfolding rates of proteins upon specific point mutations. In this work, we have systematically analyzed a set of 790 single mutants and developed a robust method for predicting protein unfolding rates upon mutations (Δlnku) in two-state proteins by combining amino acid properties and knowledge-based classification of mutants with multiple linear regression technique. We obtain a mean absolute error (MAE) of 0.79/s and a Pearson correlation coefficient (PCC) of 0.71 between predicted unfolding rates and experimental observations using jack-knife test. We have developed a web server for predicting protein unfolding rates upon mutation and it is freely available at https://www.iitm.ac.in/bioinfo/proteinunfolding/unfoldingrace.html. Prominent features that determine unfolding kinetics as well as plausible reasons for the observed outliers are also discussed.


Energy and protein metabolism in malnutrition due to nonneoplastic gastrointestinal diseases.
Although a reduction in both energy expenditure and protein turnover has been demonstrated in starved volunteers, few metabolic data are available for patients in whom malnutrition is due to nonneoplastic gastrointestinal diseases. Chronically malnourished, unstressed adult patients with nonneoplastic gastrointestinal diseases (body mass index, 15.8 +/- 2.5 kg/m2, n = 13) and healthy control subjects (n = 10) were studied in the postabsorptive state using indirect calorimetry, as well as substrate fluxes of L[1-13C]leucine, L-[2-15N]glutamine (seven patients and six controls), and D[6,6-2H2]glucose (seven patients and eight controls). Resting energy expenditure (REE) expressed in kilocalories per 24 hours was significantly lower in patients than in controls; REE expressed per unit of fat-free mass (FFM) was not significantly different in both groups. Whole-body leucine turnover, oxidation, and nonoxidative disposal rates, based on either 13C-leucine or 13C-alpha-ketoisocaproic acid (KIC) enrichments, and glucose turnover rate were not significantly different between malnourished patients and controls. Moreover, glutamine turnover was increased by 28% in malnourished patients as compared with normal volunteers (429.8 +/- 86.8 v 334.9 +/- 15.9 mumol/kg/h, P = .02). These results suggest that hypometabolic adaptation, although previously documented in starved volunteers, is not operative during states of chronic malnutrition due to gastrointestinal disease. The increase in glutamine turnover rate might represent an adaptative mechanism to malnutrition for preservation of visceral mass or function.


Virtual histology intravascular ultrasound comparison of coronary chronic total occlusions versus non-occlusive lesions.
To use virtual histology intravascular ultrasound (VH-IVUS) to assess the differences in tissue composition between chronic total occlusions (CTO) and non-occlusive lesions, especially in vivo. We compared 50 CTOs (49 patients) after guidewire crossing with 41 non-occlusive lesions (41 patients, 25 with unstable angina and 16 with stable angina). Overall, the percentage of necrotic core in the CTO segments was similar to non-occlusive lesions (35.6 vs. 38.7 %, p = 0.18 at the maximum necrotic core site; 24.2 vs. 23.7 %, p = 0.8 in mean analysis over the entire segment length). However, the percentage of dense calcium in CTO segments was less than in non-occlusive lesions (10.6 vs. 14.8 %, p = 0.025 at the maximum necrotic core site; 7.6 vs. 11.4 %, p = 0.036 in mean analysis over the entire segment length). Finally, the necrotic core/dense calcium ratio tended to be larger in the CTO segment than in non-occlusive lesions (2.94 vs. 2.35, p = 0.082 at the maximum necrotic core site; 3.05 vs. 2.03, p = 0.005 in mean analysis over the entire segment length). Overall, 42 (84 %) CTOs contained a fibroatheroma compared to 40 (97 %) of non-occulsive stenoses. The percentage of necrotic core in CTOs with a fibroathroma (39.0 %) was less than non-occlusive stenoses with a VH-thin cap fibroathroma (44.4 %, p = 0.017) and greater than non-occlusive stenoses with a VH-thick cap fibroathroma (33.5 %, p = 0.022) at maximum necrotic core site. These findings suggest that the morphological characteristics of most CTOs were similar as compared with non-CTO lesions and CTOs evolved from fibroatheromas.


Looking for new xanthine oxidase inhibitors: 3-Phenylcoumarins versus 2-phenylbenzofurans.
Overproduction of uric acid in the body leads to hyperuricemia, which is also closely related to gout. Uric acid production can be lowered by xanthine oxidase (XO) inhibitors. Inhibition of XO has also been proposed as a mechanism for improving cardiovascular health. Therefore, the search for new efficient XO inhibitors is an interesting topic in drug discovery. 3-Phenylcoumarins and 2-phenylbenzofurans are privileged scaffolds in medicinal chemistry. Their structural similarity makes them interesting molecules for a comparative study. Methoxy and nitro substituents were introduced in both scaffolds. The current study gives some insights into the synthesis and biological activity of these molecules against this important target. For the best compound of the series, the 3-(4-methoxyphenyl)-6-nitrocoumarin (4), the IC50 value, type of inhibition, cytotoxicity on B16F10 cells and ADME theoretical properties, were determined. Docking studies were also performed in order to better understand the interactions of this molecule with the XO binding pocket. This work is a preliminary screening for further design and synthesis of new non-purinergic derivatives as potential compounds involved in the inflammatory suppression, specially related to gout.


QSAR studies of the antioxidant activity of anthocyanins.
Through experimental information available from antioxidant assays of seventeen anthocyanins, and six common anthocyanidins, quantitative structure-activity relationships (QSAR) have been established in the present work. The antioxidant bioactivity has been predicted in three different lipid environments: emulsified and bulk oil (methyl linoleate) (in vitro tests) at concentrations of 50 and 250 μM, and 50 μM of the inhibitor, respectively, and in human LDL (low-density lipoprotein; "bad cholesterol") (ex vivo test) at concentrations of 2.5, 10, and 25 μM of the inhibitor. Radical scavenging activity was predicted in the assay with the 1,1-diphenyl-2-picrylhydrazyl radical (DPPH·). The QSAR models developed for each test and concentration used allowed to obtain prospective information on the constitutional and topological molecular characteristics for anthocyanin/anthocyanidin compounds. Therefore, the antioxidant activity was predicted for twenty-one compounds with unknown experimental values, leading for some of them to a favorable predicted bioactivity.


A spectrophotometric method for determination of sphingomyelinase.
A colored derivative of sphingomyelin was synthesized and used as substrate for several sphingomyelinases. The compound is N-omega-trinitrophenyl-aminolaurylsphingosylphosphorylcholine. The rate of hydrolysis of this substrate was compared to that of bovine brain sphingomyelin, labelled with tritium in the choline moiety. The following enzyme preparations were used: homogenate-less debris of brain, assayed at pH 5.0 or 7.4; a solubilized preparation derived from rat brain lysosomes, assayed at pH 5.0 and a purified enzyme of Staphylococcus aureus. With all preparations, the rates of hydrolysis of the yellow derivative were very similar to those of the brain sphingomyelin. Extracts of skin fibroblasts of normal and Niemann-Pick patients as well as amniotic cells were also used. Again, the rates of hydrolysis of the yellow derivative practically equalled those using brain sphingomyelin.


Cerebral ventricular enlargement in chronic schizophrenia: consistencies and contradictions.
A study of cerebral ventricular size measured as ventricle to brain ratio (VBR) using computerised tomographic brain scan in chronic schizophrenics provided no support for suggestions that there are significant differences between patients who fall into different clinical subtypes. We found no significant difference in VBR between patients with and without a family history of schizophrenia or between those with or without paranoid symptoms. Applying Crow's classification, contrary to expectations, Type 1 patients had significantly larger ventricles than those with 'mixed' symptomatology (both Type 1 and Type 2 features). We also applied a variety of operational criteria which attempt to define schizophrenia as a whole: of these only Schneider's first-rank symptoms (FRS) yielded conclusive results--FRS-positive patients had significantly larger mean VBR than those without such symptoms. Previously, it has been suggested that ventricular enlargement is more closely associated with 'negative' than with 'positive' symptoms.


Changes in intracranial pulse pressure amplitudes after shunt implantation and adjustment of shunt valve opening pressure in normal pressure hydrocephalus.
We have previously reported that the intracranial pulse pressure amplitudes were elevated in idiopathic normal pressure hydrocephalus (NPH) patients responding to shunt surgery. Whether or not shunt implantation or adjustment of the shunt valve opening pressure modifies the intracranial pulse pressure amplitudes in NPH patients remains to be established. This report summarises our observations. Thirteen patients with NPH (idiopathic in nine and secondary in four) are presented in whom continuous intracranial pressure (ICP) monitoring was done before and after shunt implantation. In two, ICP monitoring was also done during adjustment of shunt valve opening pressure. The mean ICP and mean ICP wave amplitude (i.e. pulse pressure amplitudes) were determined in 6-s time windows. After shunt implantation there was a fall in both mean ICP and mean ICP wave amplitude; the reduction in the two ICP parameters correlated significantly. However, mean ICP in the supine position was normal (i.e. <15 mmHg) in 12 of 13 patients before shunt placement, and remained normal after shunting. According to our criteria, the mean ICP wave amplitudes were elevated before shunting in 12 of 13 patients and became "normalised" the day after shunting in nine patients. The reduction in mean ICP wave amplitude after shunt was highly significant at the group level. Moreover, adjustment of shunt valve opening pressure modified the levels of mean ICP wave amplitudes. The present observations in 13 NPH patients indicate that shunt implantation reduces mean ICP wave amplitudes. Moreover, the level of reduction can be tailored by adjustment of the shunt valve opening pressure.


Inhibitory effects of sulfur nanoparticles on membrane lipids of Aspergillus niger: a novel route of fungistasis.
Orthorhombic (spherical; ~10 nm) and monoclinic (cylindrical; ~50 nm) sulfur nanoparticles (SNPs) were synthesized and examined for their effects on the total lipid content and desaturase enzymes of Aspergillus niger. Synthesized SNPs were characterized for size with transmission electron microscopy, elemental composition with energy dispersive X-ray spectroscopy and allotropic nature with X-ray diffraction pattern. Both the SNPs considerably reduced total lipid content of the treated fungal isolates with significant down regulation of the expression of various desaturase enzymes (linoleoyl-CoA desaturase, stearoyl-CoA 9-desaturase and phosphatidylcholine desaturase). Unusual high accumulation of saturated fatty acids with depleted lipid layer can be inferred as one of the major reasons of SNPs mediated fungistasis.


Effect of the chemosterilant bisazir on the eggs of Earias fabia stoll.
Bisazir-induced inhibition of nucleic acids and protein, and enhancement of free amino acids in the eggs of treated Earias fabia (Lepidoptera; Noctuidae) is reported. The degree of alterations in various crosses followed: T male X T female greater than UT male X T female greater than T male X UT female, which was also reflected in the weight loss of eggs. The alterations have been attributed to the abnormalities in the developing embryos due to genic imbalance in the spermatozoa, nurse cells and oocytes, ultimately leading to embryonic death.


Subjective analysis of cold thermal environments.
The present work is dedicated to the study of cold thermal environments in food distribution industrial units through a subjective assessment based on an individual questionnaire which aims to describe the working conditions of employees often exposed to cold. The survey was carried out in Portugal and the sample consists of 1575 valid responses obtained in 61 industrial units. The results show that the food distribution activity sector is characterized by a female population (78.1%) and by a young work force (63.4% of the workers are less than 35 years old). Despite the availability of cold protective clothing (52.8% of the workers indicate one garment) its characteristics require improvements. In addition almost 1/3 of the respondents consider the thermal environment cold and 79.6% of the workers report that working in the cold is harder in wintertime. The results also highlight that 37.3% of the workers report having health problems.


Porous biphasic calcium phosphate ceramics coated with nano-hydroxyapatite and seeded with mesenchymal stem cells for reconstruction of radius segmental defects in rabbits.
The osteoconduction of porous biphasic calcium phosphate (BCP) ceramics has been widely reported. In a previous study, we demonstrated that applying a nano-hydroxyapatite (nHA) coating enhances the osteoinductive potential of BCP ceramics, making these scaffolds more suitable for bone tissue engineering applications. The aim of the present study was to determine the effects of reconstructing radius defects in rabbits using nHA-coated BCP ceramics seeded with mesenchymal stem cells (MSCs) and to compare the bone regeneration induced by different scaffolds. Radius defects were created in 20 New Zealand rabbits, which were divided into four groups by treatment: porous BCP ceramics (Group A), nHA-coated porous BCP ceramics (Group B), porous BCP ceramics seeded with rabbit MSCs (Group C), and nHA-coated porous BCP ceramics seeded with rabbit MSCs (Group D). After in vitro incubation, the cell/scaffold complexes were implanted into the defects. Twelve weeks after implantation, the specimens were examined macroscopically and histologically. Both the nHA coating and seeding with MSCs enhanced the formation of new bone tissue in the BCP ceramics, though the osteoinductive potential of the scaffolds with MSCs was greater than that of the nHA-coated scaffolds. Notably, the combination of nHA coating and MSCs significantly improved the bone regeneration capability of the BCP ceramics. Thus, MSCs seeded into porous BCP ceramics coated with nHA may be an effective bone substitute to reconstruct bone defects in the clinic.


Disease progression in a patient with nonalcoholic steatohepatitis.
Nonalcoholic steatohepatitis (NASH) is a severe form of nonalcoholic fatty liver disease (NAFLD). The prevalence and clinical significance of NAFLD/NASH have been increasingly recognized in Western countries but much less known in Asian countries, including Taiwan. Here, we report the case of a 43-year-old man who had abnormal liver tests for 18 years. Retrospective evaluation of his initial clinical, laboratory and histologic findings indicated that the hepatic disorder was compatible with the diagnosis of NASH. Although his liver biochemical tests improved after taking lipid-lowering agents, a liver biopsy 17 years later demonstrated histologic progression of intralobular necroinflammation and perivenular fibrosis. These facts suggest that NASH, albeit mild and slowly progressive, indeed exists in Taiwan. After the control of chronic hepatitis B and C and westernization of the lifestyle in Taiwan, an increasing burden of NAFLD/NASH is anticipated and active prophylactic measures should be implemented.


Short communication: Ex-situ conservation in hatcheries is associated with spleen development in Lepidochelys olivacea turtle hatchlings.
Ex-situ conservation in hatcheries is a successful strategy for the recovery of sea turtle populations. However, it alters the ontogenesis of the brain and gonads, as well as body size and locomotor performance at nest emergence. Relocation to hatcheries may alter immune system development, since this depends highly on the nest environment. We hypothesized that ex-situ brooding would negatively associate with immune traits of Lepidochelys olivacea. Splenic cytoarchitecture and leukocyte quantification were used as proxies for the immune configuration. Body size, gonadal sex and sand temperature during incubation were determined. Additionally, the success of nest hatching and emergence was quantified. Linear mixed models of splenic cytoarchitecture, leucocyte proportions and body size, using sex and nest type as explanatory variables, evaluated the effects of ex-situ brooding. Generalized linear mixed models using quasibinomial distributions (log link) analyzed effects on hatching and emergence success. Hatchlings from ex-situ nests were heavier, larger and showed a greater spleen-somatic index. They showed more and better defined splenic periarteriolar lymphoid sheaths, as well as a higher proportion of heterophils but less monocytes. Moreover, ex-situ brooding increased hatching and emergence success. Sand temperatures in hatcheries favored male sex determination, while the opposite occurred for in-situ incubation. Interestingly, the immune configuration and body size were independent of sex but associated with ex-situ conservation. Greater body size promotes early hatchling survival, while better spleen development is related to a greater antibody production and a better immune response to pathogens. Altogether, the results suggest that ex-situ incubation is associated with a better immune configuration and higher survival success.


De novo phytosterol synthesis in animals.
Sterols are vital for nearly all eukaryotes. Their distribution differs in plants and animals, with phytosterols commonly found in plants whereas most animals are dominated by cholesterol. We show that sitosterol, a common sterol of plants, is the most abundant sterol in gutless marine annelids. Using multiomics, metabolite imaging, heterologous gene expression, and enzyme assays, we show that these animals synthesize sitosterol de novo using a noncanonical C-24 sterol methyltransferase (C24-SMT). This enzyme is essential for sitosterol synthesis in plants, but not known from most bilaterian animals. Our phylogenetic analyses revealed that C24-SMTs are present in representatives of at least five animal phyla, indicating that the synthesis of sterols common to plants is more widespread in animals than currently known.


Reliability of hip examination tests for femoroacetabular impingement.
To assess the interrater reliability of hip examination tests used to assess femoroacetabular impingement (FAI) among clinicians from different disciplines. Twelve subjects were examined by 9 clinicians using 12 hip tests drawn from a review of the literature and consultation with experts in hip pain and FAI. Examiners assessed both hips of each subject and were blinded to subject history. The order in which subjects were seen, the order of tests, and order of examination of the 2 hips within each subject were all randomized. Interrater reliability (IRR) for the 10 categorical tests was summarized using overall raw agreement (ORA), positive agreement (agreement on abnormal findings), and negative agreement (agreement on normal findings). An ORA of >0.75 was considered to indicate adequate reliability. For the 2 range of motion (ROM) outcomes, IRR was summarized using the median of the absolute difference (MAD) in measurements obtained by any 2 examiners on any patient. MAD reflects the "typical" difference (in degrees) between 2 raters. Adequate reliability (ORA >0.75) was achieved for 6 of the 10 hip examination tests with categorical outcomes. Positive agreement ranged from 0.35 to 0.84, while negative agreement ranged from 0.62 to 0.99. For the ROM outcomes, examiners were, on average, within 5° of each other for flexion and 7° for internal rotation. The results provide evidence that the most common hip examination tests would likely be sufficiently reliable to allow agreement between examiners when discriminating between painful FAI and normal hips in a clinical setting.


NIR persistent luminescence nanoparticles based turn-on aptasensor for autofluorescence-free determination of 17β-estradiol in milk.
Consistent exposure to 17β-estradiol through drinking water and food can cause health problems. Although many simple and sensitive fluorescence sensors of 17β-estradiol have been reported, most of them are based on fluorescence quenching test mode working in visible light range, which are inferior in anti-interference ability and quantitative range. Here, we developed a near-infrared (NIR) phosphorescence aptasensor for the detection of 17β-estradiol that has no background fluorescence. The aptasensor was based on Foster resonance energy transfer (FRET) between aptamer conjugated NIR persistent luminescence nanoparticles (PLNPs-Apt) and MoS2 nanosheets. The 17β-estradiol was quantified by the recovery of PLNPs' phosphorescence. This assay can detect 17β-estradiol in 0.5 mL samples with the LOD of 0.29 ng mL-1 and in concentrations of more than three orders of magnitude (from 0.5 ng mL-1 to 1.2 μg mL-1). This aptasensor exhibited selectivity for 17β-estradiol and was applicable in complex milk samples.


Hydrogen Gas-Mediated Deoxydehydration/Hydrogenation of Sugar Acids: Catalytic Conversion of Glucarates to Adipates.
The development of a system for the operationally simple, scalable conversion of polyhydroxylated biomass into industrially relevant feedstock chemicals is described. This system includes a bimetallic Pd/Re catalyst in combination with hydrogen gas as a terminal reductant and enables the high-yielding reduction of sugar acids. This procedure has been applied to the synthesis of adipate esters, precursors for the production of Nylon-6,6, in excellent yield from biomass-derived sources.


Functional magnetic resonance: new applications in epilepsy.
Functional magnetic resonance imaging technique is a non-invasive tool that is capable to detect the subtle hemodynamic changes produced during regional brain activation. Functional MRI applications in epilepsy patients are language lateralization and mapping, memory function assessment and localization of ictal and interictal changes. Functional MRI for language lateralization is currently used in clinical practice and provides comparable results to the intracarotid amobarbital test. In functional MRI studies for memory function assessment, results show changes in epileptic patients, but further studies are required to validate this technique for individual cases. A new application is ictal or interictal functional MRI with EEG recording that provides more detailed information about simultaneous electrographic and hemodynamic changes during interictal discharges or EEG seizures. This technique seems to have encouraging results to localize the epileptogenic area and to study propagation patterns.


Rhabdomyolysis after liver transplantation: a case report.
Our aim was to present the case of a pediatric biliary atresia patient who experienced rhabdomyolysis with severe cardiac arrhythmias associated with hyperkalemia, metabolic acidosis, and myoglobulinemia during liver transplantation. A 5-year-old girl, weighing 16.5 kg, with end-stage liver disease due to biliary atresia underwent living donor liver transplantation. A sudden onset of atrial fibrillation with rapid ventricular response was noted during the transplantation. The cardiac arrhythmia was associated with hyperkalemia, metabolic acidosis, and myoglobulinemia. Rhabdomyolysis was suspected. Hyperkalemia and metabolic acidosis were not corrected despite treatment with 10 mL of 50% glucose plus 6 U of regular insulin in 4 succeeding boluses and 110 mEq sodium bicarbonate before sending the patient to the intensive care unit. A corresponding decrease and normalization in serum potassium and correction of metabolic acidosis were noted as responses to a single dose of intravenous (20 mg) dantrolene. The patient was extubated 5 days after transplantation. The kidney function remained within normal limits during the rhabdomyolysis and the entire hospital stay. The patient was discharged 7 weeks later and is surviving with the original liver graft and satisfactory kidney function to date.


Pharmacokinetics and safety profile of tigecycline in children aged 8 to 11 years with selected serious infections: a multicenter, open-label, ascending-dose study.
Tigecycline, a broad-spectrum antibiotic used for treating serious bacterial infections in adults, may be suitable for pediatric use once an appropriate dosage is determined. The aim of this study was to assess the pharmacokinetic (PK) properties, safety profile, and descriptive efficacy of tigecycline. In this Phase II, multicenter, open-label clinical trial, children aged 8 to 11 years with community-acquired pneumonia (CAP), complicated intra-abdominal infection (cIAI), or complicated skin and skin structure infections (cSSSI) were administered tigecycline 0.75, 1, or 1.25 mg/kg. A total of 58 patients received ≥ 1 dose of tigecycline (31 boys; 44 white; mean age, 10 years; mean weight, 35 kg); 47 had data from samples available for PK analysis. The mean (SD) PK values were: C(max), 1899 (2954) ng/mL; T(max), 0.56 (0.18) hour; between-dose AUC, 2833 (1557) ng · h/mL; weight-normalized clearance, 0.503 (0.293) L/h/kg; and Vd(ss), 4.88 (4.84) L/kg. Overall clinical cure rates at test-of-cure were 94% (16/17), 76% (16/21), and 75% (15/20) in the 0.75-, 1-, and 1.25-mg/kg cohorts, respectively. The rates of protocol violations were higher in the 1- and 1.25-mg/kg groups, resulting in higher proportions of indeterminate clinical cure assessments relative to the 0.75-mg/kg cohort (19% and 15% vs 0%). The most frequent adverse event was nausea, which occurred in 50% of patients overall (29/58) and the prevalence of which was significantly higher in the 1.25-mg/kg group versus the 0.75-mg/kg group (65% vs 18%; P = 0.007). Pharmacodynamic simulations using MIC data from an ongoing microbiological surveillance trial predicted that a dosage of 1.2 mg/kg q12h would lead to therapeutic target attainment levels of up to 82% for the target AUC(0-24)/MIC ratios. A tigecycline dosage of ∼1.2 mg/kg q12h may represent the most appropriate dosage for subsequent evaluation in Phase III clinical trials in children aged 8 to 11 years with selected serious bacterial infections. ClinicalTrials.gov identifier: NCT00488345.


Accidental poisoning of a group of yearling cattle by the organophosphate insecticide trichloronat.
Eight yearling cattle were accidentally poisoned with the agricultural organophosphate insecticide trichloronat. One animal showed signs of acute organophosphate poisoning and died. The others showed signs of chronic organophosphate poisoning (organophosphate-induced delayed neuropathy) after 2-3 weeks and one animal died from renal failure at 12 weeks. High levels of trichloronat were measured in fat samples at 6 1/2 weeks. The remaining animals still showed signs of ataxia, possibly due to distal axonopathy, 1 year later but no trichloronat was detected in fat samples at this time.


Comparison of clinical results between flap-on and flap-off techniques of epithelial-laser in situ keratomileusis in correction of low to moderate myopia in eyes with thin corneas.
To prospectively compare flap-on and flap-off techniques of epithelial-laser in situ keratomileusis (epi-LASIK) for the correction of low to moderate myopia in patients with thin corneas. In this randomized, interventional study, the cohort was comprised of 88 eyes (44 patients) with myopia which underwent epi-LASIK. The epithelium was separated as a 9 mm flap with 2-4 mm nasal hinge with epikeratome (Lasatom, Gebauer Medizintechnik GmbH, Neuhausen, Germany) and the ablation was performed with the MEL 80 excimer laser (Carl Zeiss, Meditec, Jena, Germany). Thirty-eight eyes underwent flap-on Epi-LASIK where the flap was repositioned after ablation (flap-on group) and 50 eyes underwent flap-off Epi-LASIK where the epithelial flap was discarded (flap-off group). Pre- and post-operative uncorrected visual acuity (UCVA), best spectacle corrected visual acuity (BSCVA), and spherical equivalent (SE) were collected for each group. Post-operative pain, time to epithelialization, and corneal haze were also collected postoperatively. Variables were compared with the paired t-test, chi-squared test and one-way analysis of variance. Statistically significance was indicated by p < 0.05. The mean preoperative SE was -3.89 ± 1.9 diopters (D) for the flap-on group and -3.92 ± 2.17 D for the flap-off group (p = 0.96). The mean follow-up was 12 months. The mean pain score was comparable on all postoperative days except the 2nd postoperative day where the flap-on had significantly lower mean pain scores (p = 00). Time for epithelial healing was 4.39 ± 0.49 days in flap-on group and 4.64 ± 0.69 days in flap-off group (p = 0.07). There was no significant difference in the postoperative UCVA between groups (p = 0.77). Corneal haze at the end of postoperative follow-up was not different between groups (p = 0.217). There is no significant difference in the clinical outcomes between flap-on and flap-off techniques of Epi-LASIK for the correction of low to moderate myopia.


Neurofibroma: an unusual presentation.
We report neurofibroma discovered incidentally in a woman who suffered trauma to her cheek. The lesion was manifest on CT as a haematoma within a parotid tumour.


Benign Hydronephrosis and Elevated of Serum Levels of Carbohydrate Antigen CA 19-9: A Case Report.
Carbohydrate tumor-associated antigen (CA 19-9) has been shown to be upregulated in other malignant tumors including gastric, ovarian, hepatocellular, and colorectal carcinoma as well as benign diseases of the biliary track such as pancreatitis, cholangitis, and choledocholithiasis. According to the available literature, in several cases of benign hydronephrosis and in a few cases of benign renal diseases, elevated CA 19-9 has been noted. A 58-year-old Caucasian male patient was admitted in our clinic with complaints about blunt abdominal pain in the past two-month period localized in the right lumbar region and irradiating into the right inguinal area, constipation, abdominal bloating, and intermittent hematuria. The concentration of serum CA 19-9 was 3500 U/mL. Urine cytology provided no signs of abnormality. Intravenous urography visualized right-sided pyelon and ureter duplex with the defect in contrast shade of the pyelon, caused by a stag horn calculus. Contrast added computerized axial tomography of the abdomen and pelvis visualized the pyelon casted concretion spreading throughout the right pyelon, with ureterohydronephrosis with the distal block for passage of the contrast to the distal part of the ureter. There is no doubt that CA 19-9 level is occasionally elevated in patients with obstructive urolithiasis as it was in our case. In the routine medical praxis, urolithiasis should not be neglected in the differential diagnosis of elevated concentrations of CA 19-9 marker.


[Diagnosis and monitoring of endangered early pregnancies with determination of oestrone, oestradiol 17beta, oestriol, progesterone and HPL in plasma and the pregnancy test in graduated dilutions of urine (author's transl)].
Unconjugated oestrone (Oe1), oestradiol-17beta (Oe2), oestriol (Oe3), progesterone (P) and HPL in plasma were determined by radioimmunoassay and the immunological pregnancy-test in urine was carried out in 70 patients with normal pregnancy or imminent abortion from 4th-20th week of gestation. Oe2 and HPL showed the most pronounced rises, Oe3 increased especially after the first trimester. In cases with abortion symptoms and poor prognosis Oe2 and HPL gave the most reliable results concerning the endocrin function of early normal pregnancy. Oe1- and P-values in normal pregnancy did not differ so clearly from concentrations observed during normal menstrual cycles and were thus of less value. The pregnancy-test was positive (greater than 1000 IU/1) even in most cases of dead pregnancy and therefore not reliable. With increasing production of oestrogen precursors in the fetal adrenal cortex after the first trimester determination of Oe3 becomes more important. In cases with abortion symptoms in early pregnancy and subsequent normal development, plasma Oe2- and Oe3- values represented best criteria for a prognosis. -- For the diagnosis and control of the endangered early pregnancy we recommend, as a consequence of this study, determination of Oe2 up to the 13th week of pregnancy and thereafter Oe3 in maternal plasma.


Early postnatal handling alters social behavior, learning, and memory of pre- and postnatal VPA-induced rat models of autism in a context-based manner.
Early life events are known to greatly affect brain development, cortical neurogenesis, and Hypothalamic-pituitary-adrenal activity. Mainly characterized by impairment in social communication, language, and cognitive development, autism spectrum disorder (ASD) refers to a class of neuropsychiatric disorders with numerous genetic and environmental risk factors. In the early handling (EH) method, daily separation of infants from their mother, physical touching, and exposure to a new environment occur. Here, we studied the effect of EH on Social interaction, learning, and memory in rats exposed pre-or post-natally to valproic acid (VPA). Gestational VPA exposure (600 mg/kg) led to some severe autistic-like traits, more notable in the social behavior of the male sex, along with unchanged to partially altered spatial learning and memory function and reduced avoidance memory. In comparison, while causing a sex-dependent increase in spatial memory, subcutaneous injection of VPA (400 mg/kg) in infancy resulted in limited adverse autistic features, including a decrease in males' social preference, as well as reduced avoidance memory. The results indicated that neonatal handling significantly improved multiple social behavior and memory deficits in prenatally injected rats. In contrast, EH in rats receiving postnatal VPA elicited a restricted advantage on social novelty tendency; while negatively affecting some other social behavior criteria and spatial learning of males and encouraging sex-dependent repetitive behaviors in the social setting. The controversial influence of postnatal handling on juvenile rats of postnatal VPA treatment vs. prenatal VPA treatment opens up the potential for future research on the context-based consequence of early-life handling stress using different behavioral tasks and to benefit therapeutic procedures through understanding the sex- and age-specific neurobiology of short-term environmental manipulation in animal models of autism.


Genetic systems for a new approach to risk of progression of diabetic retinopathy.
To evaluate the risk of progression of diabetic retinopathy (DR) using new strategies to obtain genetic information in type 2 diabetes (T2D) based on interfering ribonucleic acid (RNA). A prospective multicentre case-control study of 132 participants was distributed into: T2D with (+DR) or without (-DR) (T2DG; n=77), and a control group (CG; n=55). After an eye examination and personal interview, tears were collected for molecular analysis (expression of microRNAs [miRNAs] (miRCURY ™ ARN Isolation Kit, Qiagen)]. Libraries, 137 vs. 140bp (GeneMapper, Applied Biosystems), were obtained in 18 samples (T2DG+DR=6; T2DG-DR=6; CG=6) by performing next-generation sequencing (NGS). SPSS 15.0 statistical program was used to perform data analysis. The mean age was 67±12 years in the T2DG vs. 55±21 years in the CG. Distribution men/women: 25/30 in T2DG vs. 51/28 in CG. A family history of DM, diet compliance, smoking, drinking and exercise, showed significant differences between groups (P<.001). A 20-25 microlitre sample of tears contained a mean of 9.42±3.30 ng/mL of purified ARN, with significant differences between T2DG/CG (P=.002) and T2DG+RD/CG (P=.004). Tear expression of miARNs in T2DG directly correlated with age/obesity/T2D duration (P<.05), and indirectly with visual acuity (P<.05). A total of 14 miRNAs related to the presence, pathogenic mechanisms and risk factors for the progression of diabetic retinopathy, were identified. We propose to use tears as a source of genetic information for DM. Specific miRNAs involved in DR development and/or progression can be used as molecular biomarkers, and based on these, for developing future biotherapies.


Electron microscopic mappings of myosin head with site-directed antibodies.
Site-directed antibodies were raised against three synthetic peptides whose sequences correspond to a region around the reactive lysine residue and two protease-sensitive regions of subfragment 1 (S1) of skeletal muscle myosin (one at the junction of the 23,000 Mr and 50,000 Mr segments, the J1 junction; and the other at the junction of the 50,000 Mr and 20,000 Mr segments of the heavy chain, the J2 junction). The antisera cross-reacted with intact myosin with titres of 5 x 10(4) (anti-J1 antiserum) and 10(4) (anti-J2 and anti-reactive lysine residue antisera). Site-specific antibodies purified by S1-Sepharose readily bound to myosin. Electron microscopic examinations of antibody-myosin complexes revealed that the J1 and J2 junctions are located 15 nm and 16 nm from the head-rod junction, respectively, while the reactive lysine residue region is 13 nm from the junction.


Solutions for the quasi-linear elliptic problems involving the critical Sobolev exponent.
In this article, we study the existence and multiplicity of positive solutions for the quasi-linear elliptic problems involving critical Sobolev exponent and a Hardy term. The main tools adopted in our proofs are the concentration compactness principle and Nehari manifold.


In Australia are people born in other countries at higher risk of road trauma than locally born people?
This study examined whether people born in other countries had higher rates of death and hospitalization due to road crashes than people born in Australia. Data on deaths that occurred in the whole of Australia between 1994 and 1997 and hospitalizations that occurred in the state of New South Wales, Australia, between 1 July 1995 and 30 June 1997 due to road crashes were analyzed. The rates of death and hospitalization, adjusted for age and area of residence, were calculated using population data from the 1996 Australian census. The study categorized people born in other countries according to the language (English speaking, non-English speaking) and the road convention (left-hand side, right-hand side) of their country of birth. Australia has the left-hand side driving convention. The study found that drivers born in other countries had rates of death or hospitalization due to road trauma equal to or below those of Australian born drivers. In contrast, pedestrians born in other countries, especially older pedestrians had higher rates of death and hospitalization due to road crashes. Pedestrians aged 60 years or more born in non-English speaking countries where traffic travels on the right-hand side of the road had risks about twice those of Australian born pedestrians in the same age group.


Phenotypic analysis of Phytophthora parasitica by using high throughput phenotypic microarray.
We studied the phenotypic characterization of Phytophthora parasitica Dastur var. nicotianae. Phenotypic characterization of the pathogen was studied to provide information for disease management program by using BIOLOG phenotype MicroArray (PM ). Using PM plates 1 to 10, 950 different phenotypic characterizations were tested. P. parasitica was able to metabolize 74% of tested carbon sources, 96% of nitrogen sources, 100% of sulfur sources, and 98% of phosphorus sources. Most informative utilization patterns for carbon sources of P. parasitica were organic acids and carbohydrates, and for nitrogen were various amino acids. The pathogen presented 285 different nitrogen pathways. It had wide range adaptabilities in osmolytes with up to 1% sodium chloride, up to 3% potassium chloride, up to 5% sodium sulfate, up to 20% ethylene glycol, up to 2% sodium formate, up to 5% urea, and up to 2% sodium lactate. It also exhibited active metabolism under pH values between 3.5 and 10, with optimal pH of around 7.0. The pathogen showed both decarboxylase and deaminase activities in the presence of various amino acids. These phenotypic characterizations of P. parasitica provided the theoretical basis for the next study of the pathogen in physiology and metabolism, and provided potential new way for tobacco black shank management.


Developing human service administrators: the educational challenge.
The author reports the results of a survey on the education of human service administrators and concludes that an interdisciplinary approach is most desirable. A model curriculum in human service administration is outlined.


A Scoping Review of the Prevalence and Correlates of Sexual Dysfunction in Adults With Substance use Disorders.
Substance use may affect sexual functioning in both men and women. Comorbid sexual dysfunction adds to the clinical burden of substance use disorders (SUD). The broad aims were to identify research conduct, types of the available evidence, and research gaps in (i) estimating the incidence, prevalence, type, and severity of sexual dysfunction in adults with SUD; (ii) exploring correlates of sexual dysfunction in SUD. We conducted systematic searches on PubMed, Google Scholar, and Embase for studies published in the English language between August 1954 and November 2020. We included prospective and cross-sectional observational studies that had examined the prevalence or incidence of any sexual dysfunction in adults of either gender with substance use disorders. Review articles and those with an exclusive focus on tobacco use disorders were excluded. The review was registered in PROSPERO. Our search identified 65 relevant articles, including five prospective studies. All the prospective studies and most of the cross-sectional studies (n = 40) were done among men and subjects with alcohol (n = 20) and opioid (n = 23) use disorders in clinical populations. Substance use and sexual dysfunction were assessed by a wide range of instruments. Prospective studies reported a prevalence of sexual dysfunction at 75% and 61% for alcohol and opioid use disorders, respectively. The prevalence of any sexual dysfunction in cross-sectional studies ranged between 15 and 100 percent. Erectile dysfunction was the most commonly studied and observed sexual dysfunction. Comorbidity and socioeconomic deprivation were consistently associated with a higher occurrence of sexual dysfunctions. We did not limit our review by the type of substances and year of publication. We adhered to the standards of conducting and reporting scoping reviews; hence, our review results should be replicable, transparent, and reliable. The wide clinical and methodological heterogeneity precluded a systematic review. Research gaps exist in women, non-clinical population, stimulants, and cannabis use disorders, and effect of treatment of SUD in sexual functioning. The quality of evidence is poor. Ghosh A, Kathiravan S, Sharma K, Mattoo SK. A Scoping Review of the Prevalence and Correlates of Sexual Dysfunction in Adults With Substance use Disorders. J Sex Med 2022;19:216-233.


The influence of age in usability testing.
The effects of age in usability testing were examined in an experiment. Sixty users from two age groups (M = 23.0 yrs, M = 58.1 yrs) operated two technical devices (keyboard-based and touchscreen-based smartphones). In addition to various performance measures (e.g. task completion time, task completion rate), several subjective measures were taken (e.g. perceived usability, affect, and workload). The results showed better performance scores for younger adults than older adults for task completion time. For older adult users there was a mismatch between usability ratings and task completion time but not between usability ratings and task completion rate. Age-related differences in the importance of speed and accuracy in task completion point to the need to consider more strongly the factor user age in usability research and practice.


Leishmania donovani vs immunity: T-cells sensitized from Leishmania of one donor may modulate their cytokines pattern on re-stimulation with Leishmania from different donor in visceral leishmaniasis.
Antimony resistance is frequently encountered during treatment of visceral leishmaniasis (VL) and the differences are well characterized by inadequate IFN-gamma dominant type-1 protection mechanisms. The part played by Leishmania parasites derived from antimony treated patients in the outcome of an immune response largely remains to be investigated. In the present study we observed that macrophages of BALB/c mice infected with antimony non-responder (SAG-NR) isolates had a greater amastigote burden than antimony responder (SAG-R) isolates. Later it was observed that antigen from SAG-NR and R L. donovani isolates elicit different cytokine responses in peritoneal blood mononuclear cells (PBMCs) from patients with VL. The production of IFN-gamma by T-cells in VL patients increased in response to Leishmania derived from responder patients but this response within same T-cells was lower when sensitized from Leishmania from a non-responder VL patient. On the other hand, IL-4 and IL-10 expression was increased when primed with parasites from non-responder VL source. Such a differential pattern of cytokine expression by the same T-cell population produced to Leishmania from different donors, needs further exploration.


Serum protein depletion by cultured post-implantation rat embryos.
Four rat serum proteins were found to be selectively depleted from the culture medium by 10 day rat embryos. These were alpha 2-macroglobulin, transferrin and 2 other proteins which had molecular weights of 132,000 and 214,000. Possibly a 5th protein, alpha 1-acid glycoprotein, was also depleted but its concentration was already low at the start of culture.


Perfusion of intact skin by a saline solution of bis(glycinato) copper(II).
The perfusion of intact cat skin by a saline solution of bis(glycinato) copper(II), labelled with Cu64, has been studied in a diffusion cell. It has been shown by counting the activity that the copper(II) complex perfuses the skin and over a period of 6-7 hr reaches the isotonic saline solution underneath. After this inaugural delay period a steady rate of penetration was maintained and after 24 hr about 1 mg of the complex perfused the skin. Skin samples, taken before and after, were fixed in buffered formalin and embedded in paraffin. Five micron sections were taken, stained with rubeanic acid and counterstained with 1% Eosin Y solution. Histological examination showed that copper was present in all layers of the perfused skin. This work is relevant to the solubility of metallic copper in human sweat and to the possible therapeutic value of the "copper bracelet".


Kinetics of replicon initiation during S phase of Chinese hamster ovary cells.
Chinese hamster ovary (CHO) cells were synchronized by a thymidine-hydroxyurea block. At different times after release from the block, cells were treated with trioxsalen and long-wavelength ultraviolet light to crosslink DNA in vivo and were labelled with [3H]thymidine for 30 min. This technique permits labelling of only the short nescent DNA fragments initiated between crosslinks. The amount of radioactivity incorporated in these fragments during the labelling period reflects the number of replicon initiation events and allows us to follow the replicon initiation pattern after removing the inhibitor. It was shown that the rate of initiation was high at the beginning of S phase and then steadily decreased.


Wireless and Closed-Loop Smart Dressing for Exudate Management and On-Demand Treatment of Chronic Wounds.
Chronic wounds have become a significant threat to people's physical and mental health and have increased the burden of social medical care. Intelligent wound dressing (IWD) with wound condition monitoring and closed-loop on-demand drug therapy can shorten the healing process and alleviate patient suffering. However, single-function wound dressings cannot meet the current needs of chronic wound treatment. Here, a wearable IWD consisting of wound exudate management, sensor monitoring, closed-loop therapy, and flexible circuit modules is reported, which can achieve effective synergy between wound exudate management and on-demand wound therapy. The dressing is attached to the wound site, and the wound exudate is spontaneously pumped into the microfluidic channel for storage. Meanwhile, the IWD can detect the state of the wound through the temperature and humidity sensor, and use this as feedback to control the liquid metal (LM) heater through a smartphone, thereby realizing the on-demand drug release from the hydrogel. In a mouse model of infected wounds, IWD accelerates wound healing by reducing inflammatory responses, promoting angiogenesis and collagen deposition.


Medium-term outcomes in cardiac transplant recipients in a single cardiac transplant center in Taiwan.
This study retrospectively investigated the outcomes of cardiac transplantation in a single medical center in Taiwan. From February 1997 to December 2005, 214 orthotopic cardiac transplantations were performed in our institution. Cumulative survival rates were compared by gender, waiting status, blood type, ischemia time, donor gender, age, and cause of brain death. The cumulative survival rates were significant different among recipient waiting status (P = .0026), blood type (P = .0376), and donor age > 40 years (P = .0260). The others parameters seem to not be different from the cumulative survival rate. There was a strong association between donors > 40 years old and increased postoperative mortality. The age of a marginal donor seemed to be > 40 years in this study.


Volatile organic compounds from endophytic fungi as innovative postharvest control of Fusarium oxysporum in cherry tomato fruits.
To assess their potential as biopesticides, the effect on the growth of phytopathogen Fusarium oxysporum of six volatile organic compounds from endophytic fungi was studied in vivo and in vitro; compounds were used both as a mixture and individually. In vivo studies were performed inoculating the pathogen into cherry tomatoes, while the in vitro antifungal effect was studied using agar dilution and gas phase methods. Also, the morphology of the hyphae exposed to these compounds was analyzed. Moreover, the possible mechanism of action of these compounds was determined by studying the respiration and cell membrane permeability. Results show that the compounds have a significant concentration-dependent antifungal effect individually and act in a synergic manner. Additionally, changes in cell membrane permeability, damage to the hyphal morphology, and an inhibitory effect on the respiration were observed. The mixture of the six compounds may be used for postharvest control of F. oxysporum in tomatoes.


A 7-year prospective study of sense of humor and mortality in an adult county population: the HUNT-2 study.
To prospectively explore the significance of sense of humor for survival over 7 years in an adult county population. Residents in the county of Nord-Trøndelag, Norway, aged 20 and older, were invited to take part in a public health survey during 1995-97 (HUNT-2), and 66,140 (71.2 %) participated. Sense of humor was estimated by responses to a cognitive (N = 53,546), social (N = 52,198), and affective (N = 53,132) item, respectively, taken from the Sense of Humor Questionnaire (SHQ). Sum scores were tested by Cox survival regression analyses applied to gender, age, and subjective health. Hazard ratios were reduced with sense of humor (continuous scale: HR = 0.73; high versus low by median split: HR = 0.50) as contrasted with increase of HR with a number of classical risk factors (e.g., cardiovascular disease: HR = 6.28; diabetes: HR = 4.86; cancer: HR = 4.18; poor subjective health: HR = 2.89). Gender proved to be of trivial importance to the effect of sense of humor in survival. Subjective health correlated positively with sense of humor and therefore might have presented a spurious relation of survival with humor, but sense of humor proved to reduce HR both in individuals with poor and good subjective health. However, above age 65 the effect of sense of humor on survival became less evident. Sense of humor appeared to increase the probability of survival into retirement, and this effect appeared independent of subjective health. Age under 65 mediated this effect, whereas it disappeared beyond this age.


Alterations in purine nucleotide metabolism during muscle differentiation in vitro.
Pathways of purine nucleotide metabolism affecting the availability of ATP in the muscle tissue were studied in differentiating rat muscle cultures. The rate of de novo purine nucleotide synthesis and of AMP deamination were found to increase markedly with cell differentiation, but the rate of IMP dephosphorylation was similarly low in both myoblasts and contracting fibers. The above differentiation-associated alterations in purine nucleotide metabolism conform with the greater need for ATP as a source of energy in the contracting myotubes.


Selective brain hypothermia attenuates focal cerebral ischemic injury and improves long-term neurological outcome in aged female mice.
This study aimed to investigate the effects of mild selective brain hypothermia on aged female ischemic mice. A distal middle cerebral artery occlusion (dMCAO) model was established in aged female mice, who were then subjected to mild selective brain hypothermia immediately after the dMCAO procedure. Neurological behavioral examinations were conducted prior to and up to 35 days post-ischemia. Infarct volume, brain atrophy, pro-inflammation, and anti-inflammation microglia/macrophages phenotype and white matter injury were evaluated by immunofluorescence staining. Correlations between neurological behaviors and histological parameters were evaluated by Pearson product linear regression analysis. Sensorimotor and cognitive function tests confirmed the protective effect of mild selective brain hypothermia in elderly female ischemic mice. In addition, hypothermia decreased the infarct volume and brain atrophy induced by focal cerebral ischemia. Furthermore, hypothermia alleviated ischemia-induced short-term and long-term white matter injury, which was correlated with behavioral deficits. Finally, hypothermia suppressed the harmful immunological response by promoting the transformation of pro-inflammatory microglia/macrophages to anti-inflammatory phenotype. This polarization was negatively correlated with neuronal loss and white matter injury. Mild selective brain hypothermia promoted long-term functional recovery by alleviating white matter damage in an aged female mouse model of ischemia.


Cardiac Resynchronization Therapy With and Without Defibrillator in a Commercial Truck Driver with Ischemic Cardiomyopathy and New York Heart Association Class III Heart Failure.
Commercial drivers warrant tighter restrictions to their driving privileges than private drivers. Patients with cardiac disease who are at risk of consciousness-impairing arrhythmias are not allowed to drive commercially. Patients with left ventricular systolic dysfunction and/or heart failure symptoms are permanently disqualified from commercial driving. A biventricular pacemaker without defibrillator can improve symptoms and mortality in selected patients with heart failure. Biventricular pacing may have antiarrhythmic effects that may reduce the added benefit of a defibrillator. Motor vehicle collisions resulting from arrhythmic events are infrequent. The interests of public safety must outweigh individual liberties when driving safety is in question.


Adult-onset Still's disease complicated by macrophage activation syndrome during pregnancy: a case-based review.
Adult-onset Still's disease is a rare, systemic inflammatory rheumatic disease characterized by recurrent fevers, arthritis, and an evanescent rash. One of the most serious hematologic derangements that can be seen with adult-onset Still's disease is macrophage activation syndrome. Macrophage activation syndrome is characterized by activation of lymphocytes, resulting in a cytokine storm and hemophagocytosis in the bone marrow, along with multi-organ failure. Adult-onset Still's disease with macrophage activation syndrome first presenting during pregnancy is exceptionally rare; here, we report two unique cases of such a presentation and review the pertinent literature. Both of our cases presented critically ill with end-organ failure, and responded to immunosuppression; fetal demise was present in one and an emergency caesarean section with a viable fetus was performed in the other patient. Maternal outcomes were favorable in both cases and both patients did well long-term with systemic therapy. Systemic immunosuppression, particularly anti-IL1 therapy, may be considered as treatment for this rare and life-threatening condition when presenting during pregnancy.


In vitro cell response to Co-containing 1,393 bioactive glass.
Cobalt ions are known to stimulate angiogenesis via inducing hypoxic conditions and hence are interesting agents to be used in conjunction with bioactive glasses (BGs) in bone tissue engineering approaches. In this work we investigated in vitro cell biocompatibility of Co releasing 1393 BG composition (in wt.%: 53SiO2, 6Na2O, 12K2O, 5MgO, 20CaO, and 4P2O5) derived scaffolds with osteoblast-like cells (MG-63) and human dermal microvascular endothelial cells (hDMECs). Cell viability, cell number and cell morphology of osteoblast-like cells in contact with particulate glass and 3D scaffolds were assessed showing good biocompatibility of 1393 reference material and with 1 wt.% CoO addition whereby 5 wt.% of CoO in the glass showed cytotoxicity. Furthermore for 1393 with 1 wt.% of CoO increased mitochondrial activity was measured. Similar observations were made with hDMECs: while 1393 and 1393 with 1 wt.% CoO were biocompatible and the endothelial phenotype was retained, 5 wt.% CoO containing BG showed cytotoxic effects after 1 week of cell culture. In conclusion, 1 wt.% Co containing BG was biocompatible with osteoblast like cells and endothelial cells and showed slightly stimulating effects on osteoblast-like cells whereas the addition of 5 wt.% CoO seems to exceed the vital therapeutic ranges of Co ions being released in physiological fluids.


Correction: A nutrient-driven tRNA modification alters translational fidelity and genome-wide protein coding across an animal genus.
[This corrects the article DOI: 10.1371/journal.pbio.1002015.].


What Determines the Quality of Rehabilitation Clinical Practice Guidelines?: An Overview Study.
The aim of the study was to determine what factors determine the quality of rehabilitation clinical practice guidelines. Six databases were searched for articles that had applied the Appraisal of Guidelines for Research & Evaluation II quality assessment tool to rehabilitation clinical practice guidelines. The 573 deduplicated abstracts were independently screened by two authors, resulting in 81 articles, the full texts of which were independently screened by two authors for Appraisal of Guidelines for Research & Evaluation II application to rehabilitation clinical practice guidelines, resulting in a final selection of 40 reviews appraising 504 clinical practice guidelines. Data were extracted from these by one author and checked by a second. Data on each clinical practice guideline included the six Appraisal of Guidelines for Research & Evaluation II domain scores, as well as the two Appraisal of Guidelines for Research & Evaluation II global evaluations. All six Appraisal of Guidelines for Research & Evaluation II domain scores were statistically significant predictors of overall clinical practice guideline quality rating; D3 (rigor of development) was the strongest and D1 (scope and purpose) the weakest (overall model P < 0.001, R2 = 0.53). Five of the six domain scores were significant predictors of the clinical practice guideline use recommendation, with D3 the strongest predictor and D5 (applicability) the weakest (overall model P < 0.001, pseudo R2 = 0.53). Quality of rehabilitation clinical practice guidelines may be improved by addressing key domains such as rigor of development.


Peptides with antimicrobial and anti-inflammatory activities that have therapeutic potential for treatment of acne vulgaris.
The pathogenesis of acne vulgaris is multifactorial involving infection of the pilosebaceous unit with Propionibacterium acnes and a cytokine-mediated inflammatory response. Five frog skin-derived antimicrobial peptides ([D4k]ascaphin-8, [G4K]XT-7, [T5k]temporin-DRa, brevinin-2GU, and B2RP-ERa), chosen for their low hemolytic activity against human erythrocytes, were assessed for their effects on the growth of clinical isolates of P. acnes and on the release of pro-inflammatory and anti-inflammatory cytokines from peripheral blood mononuclear (PBM) cells. All peptides inhibited the growth of P. acnes with the highest potency exhibited by [D4k]ascaphin-8 (minimum inhibitory concentration, MIC=3-12.5 μM). Release of TNF-α from concanavalin A (ConA)-stimulated PBM cells was significantly reduced by [D4k]ascaphin-8, [G4K]XT-7, brevinin-2GU, and B2RP-ERa (1 and 20 μg/ml) and by [T5k]temporin-DRa (20 μg/ml). Release of IFN-γ from unstimulated PBM cells was significantly reduced by [D4k]ascaphin-8 and brevinin-2GU (1 and 20 μg/ml). No peptide showed significant effects on Il-17 release. Release of the anti-inflammatory cytokines TGF-β, IL-4, and IL-10 from both unstimulated and ConA-treated PBM cells was significantly increased by [T5k]temporin-DRa and B2RP-ERa (1 and 20μg/ml). The potent activities of [D4k]ascaphin-8 and [T5k]temporin-DRa in inhibiting the growth of P. acnes and the release of pro-inflammatory cytokines, and in stimulating the release of anti-inflammatory cytokines suggest a possible therapeutic role in the treatment of acne vulgaris.


Ferric citrate and apo-transferrin enable erythroblast maturation with β-globin from hemogenic endothelium.
Red blood cell (RBC) generation from human pluripotent stem cells (PSCs) offers potential for innovative cell therapy in regenerative medicine as well as developmental studies. Ex vivo erythropoiesis from PSCs is currently limited by the low efficiency of functional RBCs with β-globin expression in culture systems. During induction of β-globin expression, the absence of a physiological microenvironment, such as a bone marrow niche, may impair cell maturation and lineage specification. Here, we describe a simple and reproducible culture system that can be used to generate erythroblasts with β-globin expression. We prepared a two-dimensional defined culture with ferric citrate treatment based on definitive hemogenic endothelium (HE). Floating erythroblasts derived from HE cells were primarily CD45+CD71+CD235a+ cells, and their number increased remarkably upon Fe treatment. Upon maturation, the erythroblasts cultured in the presence of ferric citrate showed high transcriptional levels of β-globin and enrichment of genes associated with heme synthesis and cell cycle regulation, indicating functionality. The rapid maturation of these erythroblasts into RBCs was observed when injected in vivo, suggesting the development of RBCs that were ready to grow. Hence, induction of β-globin expression may be explained by the effects of ferric citrate that promote cell maturation by binding with soluble transferrin and entering the cells.Taken together, upon treatment with Fe, erythroblasts showed advanced maturity with a high transcription of β-globin. These findings can help devise a stable protocol for the generation of clinically applicable RBCs.


Contact angle and detachment energy of shape anisotropic particles at fluid-fluid interfaces.
The three phase contact angle of particles, a measure of its wettability, is an important factor that greatly influences their behaviour at interfaces. It is one of the principal design parameters for potential applications of particles as emulsion/foam stabilizers, functional coatings and other novel materials. In the present work, the effect of size, shape and surface chemistry of particles on their contact angle is investigated using the gel trapping technique, which facilitates the direct visualization of the equilibrium position of particles at interfaces. The contact angle of hematite particles of spherocylindrical, peanut and cuboidal shapes, hematite-silica core-shell and silica shells is reported at a single particle level. The spherocylindrical and peanut shaped particles are always positioned with their major axis parallel to the interface. However, for cuboidal particles at air-water as well as decane-water interfaces, different orientations namely - face-up, edge-up and the vertex-up - are observed. The influence of gravity on the equilibrium position of the colloidal particles at the interface is studied using the hematite-silica core-shell particles and the silica shells. The measured contact angle values are utilized in the calculations of the detachment and surface energies of the hematite particles adsorbed at the interface.


Impact on Physicochemical Composition and Antioxidant Activity of the Wild Edible Mushroom Cyttaria espinosae Subjected to Drying.
The wild edible mushroom 'digüeñe' (Cyttaria espinosae) is a mushroom with interesting flavor and nutritional properties, but highly perishable with a consumption very limited, generating inevitable economic losses for their producers. Because of that, the aim of this study was the evaluation of two drying methods (hot-air drying and freeze-drying) in its bioactive and physicochemical composition for its preservation. The best performances of antioxidant capacity were 62 μmol TE g-1 (ORAC) and 14.21 μmol TE g-1 (DPPH), being higher compared with other vegetable sources. Its protein (5.33 %), fiber (5.58 %) and K (2.82 mg g-1 ) content also stands out. β-Carotene retention was 35.02 and 23.25 % after freeze drying and hot-air, respectively. Freeze-drying seems a good strategy for digüeñe, bioactive compounds, color and structure were better maintained. This work represents the first characterization of dehydrated digüeñe, constituting approaches for its valorization and use in the food industry.


Risk factors of fecal carriage with extended-spectrum β-lactamase-producing Enterobacteriaceae in hospitalized patients.
Colonization of multidrug-resistant microorganisms is a potential source of cross transmission and subsequent infection. Screening of extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) is not yet a routine practice in most hospitals. The purpose of this study was to investigate the risk factors of fecal carriage of ESBL-E in hospitalized patients. A total of 174 samples (from 97 patients) was collected. Screening test was accomplished using commercial chromogenic medium (ChromID ESBL agar; bioMérieux, Marcy l'Etoile, France). The presence of ESBLs of grown isolates was confirmed by Clinical and Laboratory Standards Institute guidelines. Analysis of the risk factors of ESBL-E carriage was performed by multivariate logistic regression. Among the risk factors, multivariate analysis demonstrated that prolonged hospitalization (≥90 days) (odds ratio [OR], 6.76; 95% confidence interval [CI]: 1.17-39.23; P = .033), prolonged intensive care unit stay (≥14 days) (OR, 4.78; 95% CI: 1.70-13.42; P = .003), and methicillin-resistant Staphylococcus aureus carriage (OR, 3.29; 95% CI: 1.20-9.01; P = .020) were found to be significantly associated with fecal carriers. This study clarified the characteristics and the risk factors of hospitalized patients carrying ESBL-E. In addition, the cost-benefit of routine screening in addition to methicillin-resistant Staphylococcus aureus and vancomycin-resitant Enterococci needs to be assessed.


Structural basis for the inhibition of human angiotensin-1 converting enzyme by fosinoprilat.
Human angiotensin I-converting enzyme (ACE) has two isoforms, somatic ACE (sACE) and testis ACE (tACE). The functions of sACE are widespread, with its involvement in blood pressure regulation most extensively studied. sACE is composed of an N-domain (nACE) and a C-domain (cACE), both catalytically active but have significant structural differences, resulting in different substrate specificities. Even though ACE inhibitors are used clinically, they need much improvement because of serious side effects seen in patients (~ 25-30%) with long-term treatment due to nonselective inhibition of nACE and cACE. Investigation into the distinguishing structural features of each domain is therefore of vital importance for the development of domain-specific inhibitors with minimal side effects. Here, we report kinetic data and high-resolution crystal structures of both nACE (1.75 Å) and cACE (1.85 Å) in complex with fosinoprilat, a clinically used inhibitor. These structures allowed detailed analysis of the molecular features conferring domain selectivity by fosinoprilat. Particularly, altered hydrophobic interactions were observed to be a contributing factor. These experimental data contribute to improved understanding of the structural features that dictate ACE inhibitor domain selectivity, allowing further progress towards designing novel 2nd-generation domain-specific potent ACE inhibitors suitable for clinical administration, with a variety of potential future therapeutic benefits. DATABASE: The atomic coordinates and structure factors for nACE-fosinoprilat and cACE-fosinoprilat structures have been deposited with codes 7Z6Z and 7Z70, respectively, in the RCSB Protein Data Bank, www.pdb.org.


Diagnosis and surgical treatment of primary aldosteronism in pregnancy: a case report.
Aldosterone-producing adrenal adenomas are rare, especially during pregnancy. We report a patient who presented in the early second trimester, was diagnosed with primary aldosteronism, and was treated successfully by adrenalectomy. A 17-year-old black nulliparous woman was found to have a blood pressure (BP) of 150/82 mmHg when she registered for prenatal care at 14 weeks' gestation. Initial laboratory assessment revealed a markedly diminished serum potassium level of 2.1 mmol/L. Further laboratory evaluation detected decreased random plasma renin activity and an elevated aldosterone level. Magnetic resonance imaging revealed a 2-cm right adrenal lesion. She was diagnosed with an adrenal adenoma and successfully underwent an adrenalectomy at 17 weeks' gestation. Postoperatively, her BP and serum potassium level normalized. She spontaneously delivered a normal male infant at term. Although primary hyperaldosteronism is a rare clinical entity, it must be considered when hypertension and hypokalemia are present concurrently. Antepartum medical management can be difficult, often resulting in poor obstetric outcome. Surgery in the second trimester is an effective option.


Effect of number of diffusion encoding directions in neonatal diffusion tensor imaging using Tract-Based Spatial Statistical analysis.
Diffusion tensor imaging (DTI) has been used to study the developing brain in early childhood, infants and in utero studies. In infants, number of used diffusion encoding directions has traditionally been smaller in earlier studies down to the minimum of 6 orthogonal directions. Whereas the more recent studies often involve more directions, number of used directions remain an issue when acquisition time is optimized without compromising on data quality and in retrospective studies. Variability in the number of used directions may introduce bias and uncertainties to the DTI scalar estimates that affect cross-sectional and longitudinal study of the brain. We analysed DTI images of 133 neonates, each data having 54 directions after quality control, to evaluate the effect of number of diffusion weighting directions from 6 to 54 with interval of 6 to the DTI scalars with Tract-Based Spatial Statistics (TBSS) analysis. The TBSS analysis was applied to DTI scalar maps, and the mean region of interest (ROI) values were extracted using JHU atlas. We found significant bias in ROI mean values when only 6 directions were used (positive in fractional anisotropy [FA] and negative in fractional anisotropy [MD], axial diffusivity [AD] and fractional anisotropy [RD]), while when using 24 directions and above, the difference to scalar values calculated from 54 direction DTI was negligible. In repeated measures voxel-wise analysis, notable differences to 54 direction DTI were observed with 6, 12 and 18 directions. DTI measurements from data with at least 24 directions may be used in comparisons with DTI measurements from data with higher numbers of directions.


Pharmacodynamics and pharmacokinetics of oral pirmenol.
The efficacy, pharmacokinetics, and pharmacodynamics of pirmenol, a class Ia antiarrhythmic agent, were studied in patients with frequent symptomatic premature ventricular complexes (PVCs). Pirmenol was given every 12 hours to eight patients in a dose-ranging protocol, and median PVC suppression of 94% (range 72% to 100%) was achieved. The median effective pirmenol dose was 300 mg/day (range 200 to 500 mg/day), and mean (+/- SD) trough plasma pirmenol concentration at the effective dose was 0.98 +/- 0.29 micrograms/ml. The mean half-life of elimination was 10.5 +/- 2 hours. There was considerable overlap among patients with respect to plasma pirmenol concentration and times at which PVC frequency returned to 25%, 50%, and 75% of baseline during drug washout trials. Altering pirmenol's dose interval (while maintaining a constant daily dose) from 12 to 6 hours did not improve drug efficacy. Pirmenol was given to seven patients for long-term therapy (24 to 44 months). Median PVC suppression at 24 months was 70%. Pirmenol is safe and well tolerated, and it can be administered twice daily for PVC suppression.


Differential elimination of radioiodine from single- and double-stranded 125I-DNA.
[125I]Iododeoxyuridyl residues in heat-denatured I-DNA underwent deiodination in solutions of sodium bisulfite. In 0.5 M bisulfite containing 1 M trimethylamine, 92% of the iodine was released from denatured DNA in 2.5 min at 60 degrees C. By contrast iodine in native I-DNA remained stably bound under the same condition. The deiodination of [125I]I-deoxyuridine residues incorporated by enzymatic copying of oligomers 60 nucleotides in length was found to be pairing dependent. While 125I in iododeoxyuridine paired to deoxyadenosine in a complementary oligomer remained approximately equal to 90% covalently bound in the presence of 1 M bisulfite at 60 C, when the iodouridylate residue was mismatched greater than 96% was eliminated in 4 min. The presence of mismatches in the two nucleotides adjacent to a [125I]iododeoxyuridine/deoxyadenosine pair increased the elimination of iodine. The results indicate that the deiodination reaction might be applied for the rapid detection of variations in natural DNA sequences.


Visualization of Results from Systems Genetics Studies in Chromosomal Context.
This chapter describes methods currently available for visualizing results from systems genetics experiments. Here, we abstract from the statistical methods used for genetic mapping, which are dependent on the specific resource being used, i.e. F2, RILs, or outbred populations among others. We use a public dataset with results from a mouse eQTL experiment for three examples of visualization: genome-wide dot plots of marker-by-gene association, karyotype-like plots, and circos plots. Dot plots give a first overview of the results from eQTL mapping, allowing detecting genome-wide patterns of cis- and trans-genetic association to transcription level. Karyotype-like plots provide chromosomal context and allow integrating multiple tracks of information in a single plot. Circos plots can, in addition, display long-range interactions to provide an overview of genetic connectivity at the genome level. All examples are developed and explained using R code, an open-source language with powerful statistical and graphical capabilities. The principles reviewed here, however, can be applied with other software options, organisms, and to any type of molecular phenotype that can be assigned to a genomic position.


Mechanistic Insights into the Multiple Activities of the Rad5 Family of Enzymes.
The budding yeast protein Rad5 is highly conserved among eukaryotes. Rad5 and its orthologs including helicase like transcription factor (HLTF) and SNF2 histone linker PHD RING helicase (SHPRH) in humans constitute a unique family of enzymes that play critical roles in the cellular response to DNA replication stresses. The function of the Rad5 family of enzymes is fulfilled by their multiple activities, including ubiquitin ligase, replication fork regression activities and others. Herein, we review recent studies that provided mechanistic insights into their multiple activities and their coordination.


One-step purification of mouse monoclonal antibodies by mass ion exchange chromatography on Zetaprep.
A rapid one-step purification procedure was developed to isolate mouse monoclonal antibodies present in the growth medium of hybridoma cultures. The procedure, based on a sulfopropyl mass ion exchange chromatography, yields considerable amounts of purified monoclonal antibody. The immunoglobulins are essentially free of transferrin and albumin, as determined by SDS-polyacrylamide gel electrophoresis.


Proteolytic activity in the digestive tract of several species of fish with different feeding habits.
Proteolytic activity in the gut contents of two cichlids and six cyprinids from an artificial basin in Sri Lanka was measured using a simple film strip method. This comparative study contributes to our general knowledge of digestion in herbivorous fish: 1) Specific proteolytic activity (per ml of gut content) is lower in herbivorous than in omnivorous and carnivorous species. 2) Specific proteolytic activity is negatively correlated with the relative length of the gut, but the time of exposure of ingested food to proteolytic enzymes rises with increasing gut length. This results in more intensive proteolytic digestion in herbivorous fish. 3) Proteolytic enzymes seem to be "reabsorbed" in the hindgut of the fish. The effectiveness of this mechanism rises up to a relative gut length of 2.5-3.0. However, the small species Amblypharyngodon melletinus does not fit this relationship.


Blockade of spinal nitric oxide synthase on blood pressure variability and hepatic microcirculation.
We previously demonstrated that intrathecal administration of N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, increased systemic arterial blood pressure in a dose-dependent manner in rats. The aim of the study was to investigate the participation of autonomic nervous system on L-NAME-induced hypertension and also illuminate its effects on hepatic microcirculation in rats. Eight Spraque-Dawley rats were used and initially anesthetized with ketamine 120 mg/kg, intraperitoneally supplemented by intravenous infusion of ketamine at 30 mg/kg/h for maintenance. Surgical preparations included cannulations of right femoral artery and vein to obtain systemic arterial pressure signals and administer anesthetic drug. A mini-laparotomy was made to facilitate the insertion of a microdialysis probe and attachment of a laser Doppler probe to the middle lobe of the liver. On experiment, L-NAME was administered via the previously placed intrathecal catheter at 0, 0.37, 0.74, and 1.48 micromol in sequence at a 2-h interval. The results showed that the arterial blood pressure increased in a dose-dependent manner. By the same token, the power density of very low frequency (VLF) also increased. The low frequency (LF): high frequency (HF) ratio shifted toward parasympathetic dominance. Blood flow to the liver was unchanged except slightly decreased in the animals receiving 0.37 micromol. The levels of monoethylglycinexylidide (MEGX), an index of hepatic metabolism, were unchanged throughout the experiment. We concluded that the blockade of spinal nitric oxide synthase by intrathecal administration of L-NAME significantly increased vasomotor tone in a dose-dependent manner and as a consequence induced a reflex sympathetic inhibition. Hepatic microcirculation was stable with the applied doses.


Inhibition of orexin-1/hypocretin-1 receptors inhibits yohimbine-induced reinstatement of ethanol and sucrose seeking in Long-Evans rats.
Previous studies have shown that orexin-1/hypocretin-1 receptors play a role in self-administration and cue-induced reinstatement of food, drug, and ethanol seeking. In the current study, we examined the role of orexin-1/hypocretin-1 receptors in operant self-administration of ethanol and sucrose and in yohimbine-induced reinstatement of ethanol and sucrose seeking. Rats were trained to self-administer either 10% ethanol or 5% sucrose (30 min/day). The orexin-1 receptor antagonist SB334867 (0, 5, 10, 15, 20 mg/kg, i.p.) was administered 30 min before the operant self-administration sessions. After these experiments, the operant self-administration behaviors were extinguished in both the ethanol and sucrose-trained rats. Upon reaching extinction criteria, SB334867 (0, 5, 10 mg/kg, i.p.) was administered 30 min before yohimbine (0 or 2 mg/kg, i.p.). In a separate experiment, the effect of SB334867 (0, 15, or 20 mg/kg, i.p.) on general locomotor activity was determined using the open-field test. The orexin-1 receptor antagonist, SB334867 (10, 15 and 20 mg/kg) decreased operant self-administration of 10% ethanol but not 5% sucrose self-administration. Furthermore, SB334867 (5 and 10 mg/kg) significantly decreased yohimbine-induced reinstatement of both ethanol and sucrose seeking. SB334867 did not significantly affect locomotor activity measured using the open-field test. The results suggest that inhibition of OX-1/Hcrt-1 receptors modulates operant ethanol self-administration and also plays a significant role in yohimbine-induced reinstatement of both ethanol and sucrose seeking in rats.


Using routine data for quality assessment in NeuroNet telestroke care.
Systematic clinical trials are often unavailable to evaluate and optimize operational telestroke networks. In a complementary approach, readily available routine clinical data were analyzed in this study to evaluate the effect of a telestroke network over a 4-year period. Routine clinical data from the HELIOS hospital information system were compared before and after implementation of the NeuroNet concept, including neurologic acute stroke teleconsultations, standard operating procedures, and peer review quality management in 3 hospital cohorts: 5 comprehensive stroke centers, 5 NeuroNet hospitals, and 5 matched control hospitals. During the study period, the rate of thrombolytic therapy increased by 4.8% in NeuroNet hospitals, while ischemic stroke in-hospital mortality decreased (relative risk reduction ~29% in NeuroNet and control hospitals). The odds ratio for thrombolytic therapy in comprehensive stroke centers compared to NeuroNet hospitals was reduced from 3.7 to 1.3 between 2006 and 2009. Comprehensive stroke care coding according to German Diagnosis Related Groups definitions increased by 45% in NeuroNet (P < .0001) and by 18% in control hospitals. Routine clinical data on in-hospital mortality, the rate of thrombolytic therapy, and comprehensive stroke care coding reflect different aspects of acute stroke care improvement related to the implementation of the telemedical NeuroNet concept and unified quality management (standard operating procedure teaching concept, peer review process). Similar evaluation processes could contribute to quality monitoring in other telestroke networks.


Type C brachydactyly transmitted through four generations.
We report a type C brachydactyly transmitted through four generations, with incomplete penetrance and feet abnormalities in the proposita.


Cardiovascular diseases and psychosocial factors at work.
Besides the 'classic' cardiovascular risk factors (high blood pressure, dyslipidaemia, metabolic syndrome and diabetes), the work environment is playing an increasingly significant role in cardiovascular morbidity and mortality. Several elements contribute to the effect of the work environment: physical factors, chemical factors, shift work and psychosocial factors. The effects of psychosocial factors on the aetiology and progression of cardiovascular disease have been confirmed by several studies. Identification of these work-related psychosocial factors must be taken into account when evaluating cardiovascular risk factors, in order to ensure better prevention.