Chromatin Immunoprecipitation (ChIP) of Histone Modifications from Saccharomyces cerevisiae.
Histone post-translational modifications (PTMs), such as acetylation, methylation and phosphorylation, are dynamically regulated by a series of enzymes that add or remove these marks in response to signals received by the cell. These PTMS are key contributors to the regulation of processes such as gene expression control and DNA repair. Chromatin immunoprecipitation (chIP) has been an instrumental approach for dissecting the abundance and localization of many histone PTMs throughout the genome in response to diverse perturbations to the cell. Here, a versatile method for performing chIP of post-translationally modified histones from the budding yeast Saccharomyces cerevisiae (S. cerevisiae) is described. This method relies on crosslinking of proteins and DNA using formaldehyde treatment of yeast cultures, generation of yeast lysates by bead beating, solubilization of chromatin fragments by micrococcal nuclease, and immunoprecipitation of histone-DNA complexes. DNA associated with the histone mark of interest is purified and subjected to quantitative PCR analysis to evaluate its enrichment at multiple loci throughout the genome. Representative experiments probing the localization of the histone marks H3K4me2 and H4K16ac in wildtype and mutant yeast are discussed to demonstrate data analysis and interpretation. This method is suitable for a variety of histone PTMs and can be performed with different mutant strains or in the presence of diverse environmental stresses, making it an excellent tool for investigating changes in chromatin dynamics under different conditions.


Italian guidelines for the management and treatment of neonatal cholestasis.
Hyperbilirubinemia is a frequent condition affecting newborns during the first two weeks of life and when it lasts more than 14 days it is defined as prolonged jaundice. This condition requires differential diagnosis between the usually benign unconjugated hyperbilirubinemia and the pathological conjugated hyperbilirubinemia, that is mainly due to neonatal cholestasis. It is important that the diagnosis of neonatal cholestasis be well-timed to optimize its management, prevent worsening of the patient's outcome, and to avoid premature, painful, expensive, and useless tests. Unfortunately, this does not always occur and, therefore, the Task Force on Hyperbilirubinemia of the Italian Society of Neonatology presents these shared Italian guidelines for the management and treatment of neonatal cholestasis whose overall aim is to provide a useful tool for its assessment for neonatologists and family pediatricians.


[A historical perspective of Ebola virus].
The 2014 Ebola fever outbreak was the first of its kind in West Africa. This epidemic, affecting multiple countries, by far exceeded any previous outbreak in case counts and geographical spread. But Ebola viruses are not new to Africa, as they have occurred in epidemic proportion in the central part of the continent since 1976. The objective of this article is to compare previous outbreaks with the ongoing epidemic in an effort to identify some of the factors which have con-tributed to the size and scope of the existing outbreak.


[Genetics of multiresistant MRSA].
Since the introduction of methicillin, methicillin resistant S. aureus (MRSA) was appeared in 1961 in England. MRSA produces specific penicillin-binding protein PBP2' which shows very low affinity to most of beta-lactam antibiotics. The region around mecA is called additional DNA or mec DNA and is thought to be extraspecies origin. From the study of mec DNA, it was revealed that there were three types of mec DNA. In the case of mec DNA of N315, a transposon Tn554 which encoded erythromysin and spectinomycin resistance, and a plasmid pUB110 which encoded resistance to kanamycin, tobramycin, and bleomycin were integrated. MRSA became resistant to many antibiotics during a few years which were effective at the time of introduction, such as, carbapenems, new quinolones, and minocycline. MRSA has changed their properties by obtaining resistance-genes or generating mutations on its chromosomal DNA.


Elevated plasma cotinine is associated with an increased risk of developing IBD, especially among users of combusted tobacco.
Smoking has previously been associated with inflammatory bowel disease (IBD), but no study has reported on cotinine, an objective, biochemical measure of tobacco use. We aimed at testing the hypothesis that cotinine levels among healthy subjects are associated with an increased risk of developing IBD in later life. We analysed plasma cotinine and evaluated corresponding lifestyle questionnaires that included tobacco habits in subjects (n = 96) who later developed late-onset IBD (70 ulcerative colitis (UC) and 26 Crohn's disease (CD)) and in sex and age-matched controls (n = 191). Patients who later developed IBD had significantly higher plasma cotinine levels compared to controls. In multivariable analysis, higher log-cotinine was associated with a higher risk of developing IBD (OR 1.34 (95% CI 1.01-1.63)). After stratifying for time to diagnosis, the association was only significant in subjects with shorter time (< 5.1 years) to diagnosis (OR 1.45 (1.09-1.92)). The findings were similar for UC- and CD-cases, but did not reach statistical significance in CD-cases. Although plasma cotinine concentrations were higher in snuff users compared to combusted tobacco users, no increase in the risk of IBD and lower risk of developing IBD among subjects with shorter time (< 5.1 years) to diagnosis was seen among snuff users. Cotinine, a biomarker of tobacco use, is associated with increased risk of developing late-onset IBD in general, and UC in particular. No increased risk among snuff users indicates that other components in combusted tobacco than nicotine may be involved in the pathogenesis of IBD among smokers.


The role of the stem cell leukemia (SCL) gene in hematopoietic and endothelial lineage specification.
Anatomical observations made at the beginning of the twentieth century revealed an intimate association between the ontogeny of blood and endothelium and led to the hypothesis of a common cell of origin termed the hemangioblast. However, the precise nature of the cellular intermediates involved in the development of both lineages from uncommitted precursors to mature cell types is still the subject of ongoing studies, as are the molecular mechanisms driving this process. There is clear evidence that lineage-restricted transcription factors play a central role in the genesis of mature lineage committed cells from multipotent progenitors. Amongst these, the basic helix-loop-helix (bHLH) family is of key importance for cell fate determination in the development of the hematopoietic system and beyond. This article will review the current evidence for the common origin of blood and endothelium, focusing on the function of the bHLH protein encoded by the stem cell leukemia (SCL) gene, and its role as a pivotal regulator of hematopoiesis and vasculogenesis.


Circulating levels of angiogenesis-related growth factors in breast cancer: A study to profile proteins responsible for tubule formation.
The present study exploited a versatile in vitro endothelial cell/fibroblast co-culture cell system to investigate the association between angiogenesis and breast cancer by comparing the capacity of plasma from women with breast cancer and age-matched controls, to influence tubule formation and modulate angiogenesis in vitro, and to identify plasma circulating factors which might be responsible. Plasma from women with breast cancer (n=8) (added on day 7 after co-culture establishment) significantly increased tubule formation by 57% (P<0.01) when compared to cultures grown in culture medium lacking in vascular endothelial growth factor (VEGF) and fetal bovine serum (FBS), whereas plasma from controls (n=8) did not. Higher levels of VEGF, tumour necrosis factor-α (TNFα) and interleukin (IL)-6, but not leptin, were observed in plasma samples of the breast cancer group compared to the control group (n=20 in each group). In independent experiments, the effects of VEGF, TNFα, IL-6 and leptin were assessed and it was found that tubule formation was differentially affected whether these inflammatory cytokines or adipokines were added individually or in combination to the co-culture system. Using Proteome Profiler human angiogenesis array kits, 12 out of 55 angiogenesis-related proteins were differentially expressed in plasma from the breast cancer group compared to the control group. Pro-angiogenic proteins included: amphiregulin, artemin, coagulation factor III, fibroblast growth factor (FGF) acidic, GDNF, IL-8, macrophage inflammatory protein (MIP)-1α, platelet derived growth factor-AB/platelet derived growth factor-BB (PDGF-AB/PDGF-BB) and VEGF, whereas anti-angiogenic proteins were: angiopoietin-2, serpin F1 and serpin B5. In addition, FGF acidic was further identified as differentially expressed, with increased expression, when plasma samples from the normal and cancer groups, which induced an increase in tubule formation, were compared to one another. In conclusion, the present study identified angiogenesis-related proteins circulating in the serum of women with breast cancer that are likely to facilitate the growth and metastasis of breast cancer, in part through their influence on tubule formation, and, therefore, may be potential targets for new cancer therapies.


Toxicology and carcinogenesis studies of methyl trans-styryl ketone (CAS NO 1896-62-4) in F344/N rats and B6C3F1 mice (feed and dermal studies).
Methyl trans-styryl ketone is used as a synthetic flavoring agent and a fragrance additive in food and personal care products. Methyl trans-styryl ketone was nominated for study by the National Cancer Institute due to widespread human exposure as a flavoring and fragrance additive, positive results in the Ames/Salmonella assay and the mouse lymphoma L5178Y/tk+/- assay, and as a representative of the α,β-unsaturated ketone chemical class. Male and female F344/N rats and B6C3F1 mice received methyl trans-styryl ketone (98.6% pure) in feed for 3 months and dermally for 3 months or 2 years. Genetic toxicology studies were conducted in Salmonella typhimurium, Escherichia coli, and mouse peripheral blood erythrocytes. Two-year studies were conducted to provide data for assessment of possible toxicity due to exposure to methyl trans-styryl ketone. The dermal route was chosen since this is the route for highest human exposure and due to studies demonstrating systemic exposure following dermal application to methyl trans-styryl ketone. 3-MONTH FEED STUDY IN RATS Groups of 10 male and 10 female rats were fed diets containing 0%, 0.025%, 0.05%, 0.1%, 0.2%, or 0.4% methyl trans-styryl ketone (equivalent to average daily doses of approximately 18, 36, 72, 145, or 290 mg methyl trans-styryl ketone/kg body weight to males and 19, 38, 77, 150, or 300 mg/kg to females) for 14 weeks. Groups of 10 male and 10 female clinical pathology rats were fed the same concentrations for 24 days. All core study rats survived to the end of the study. Final mean body weights of males and females receiving 0.4% and mean body weight gains of males receiving 0.4% were significantly less than those of the controls. Feed consumption by exposed groups was similar to that by the controls. Clinical findings included diarrhea and hyperactivity in males and females. Results of sperm motility and vaginal cytology evaluations indicated methyl trans-styryl ketone is unlikely to be a reproductive toxicant in male rats; however, it exhibits potential for reproductive toxicity in female rats based upon an increased probability of extended diestrus at the highest exposure concentration. In all exposed groups of males, there were treatment-related increased incidences of goblet cell hyperplasia of the respiratory epithelium of the nose and nephropathy of the kidney. In females, there was an increased incidence of goblet cell hyperplasia of the respiratory epithelium of the nose in the group receiving 0.4%. 3-MONTH FEED STUDY IN MICE: Groups of 10 male and 10 female mice were fed diets containing 0%, 0.025%, 0.05%, 0.1%, 0.2%, or 0.4% methyl trans-styryl ketone (equivalent to average daily doses of approximately 55, 110, 220, 400, or 750 mg/kg to males and 50, 100, 200, 350, or 600 mg/kg to females) for 14 weeks. One male receiving 0.2% and one control female died before the end of the study. Mean body weights of males and females receiving 0.4% were significantly less than those of the controls. Feed consumption by exposed groups was similar to that by the controls. Hyperactivity in both sexes was the only clinical finding. Results of sperm motility and vaginal cytology evaluations indicated methyl trans-styryl ketone is unlikely to be a reproductive toxicant in male mice; however, it exhibits potential for reproductive toxicity in female mice based upon an increased probability of extended diestrus at the lowest and the highest exposure concentrations. There were significantly increased incidences of olfactory epithelial atrophy of the nose in males and females receiving 0.4%. 3-MONTH DERMAL STUDY IN RATS: Groups of 10 male and 10 female rats were dermally administered 0, 22, 44, 87.5, 175, or 350 mg methyl trans-styryl ketone/kg body weight in 95% ethanol, 5 days per week for 14 weeks. Groups of 10 male and 10 female clinical pathology rats were administered the same doses for 23 days. All rats survived to the end of the study. Mean body weights of 175 and 350 mg/kg males were significantly less than that of the vehicle controls. Clinical findings in groups administered 175 or 350 mg/kg included dermal irritation, thickened skin, and ulceration at the site of application. Results of sperm motility and vaginal cytology evaluations indicated methyl trans-styryl ketone is unlikely to be a reproductive toxicant in male or female rats at the doses used in this study. Histologically, there were significantly increased incidences of epidermal hyperplasia, hyperkeratosis, chronic active inflammation, epidermal necrosis, and sebaceous gland hypertrophy in the skin at the site of application in males and/or females. There were significantly increased incidences of goblet cell hyperplasia of the nose in 350 mg/kg males and 22, 175, and 350 mg/kg females. 3-MONTH DERMAL STUDY IN MICE Groups of 10 male and 10 female mice were dermally administered 0, 87.5, 175, 350, 700, or 1,400 mg methyl trans-styryl ketone/kg body weight in 95% ethanol, 5 days per week for 13 weeks. All mice in the 700 and 1,400 mg/kg groups were sacrificed moribund before the end of the study. The final mean body weights of surviving groups of dosed males and females were similar to those of the vehicle controls; however, the mean body weight gains of the 175 mg/kg groups were significantly less than those of the vehicle controls. Clinical findings at the site of application included dermal irritation in 350 mg/kg males and crust formation in all 700 and 1,400 mg/kg mice except one female. Results of sperm motility and vaginal cytology evaluations indicated methyl trans-styryl ketone is unlikely to be a reproductive toxicant in male or female mice at the doses used in this study. There were treatment-related increased incidences of epidermal hyperplasia, hyperkeratosis, chronic active inflammation, epidermal necrosis, sebaceous gland hypertrophy, and hair follicle hyperplasia in the skin at the site of application in males and females. There were increased incidences of olfactory epithelial atrophy of the nose in groups of males and females administered 350 mg/kg or greater. 2-YEAR DERMAL STUDY IN RATS: Groups of 50 male and 50 female rats were dermally administered 0, 10, 30, or 90 mg methyl trans-styryl ketone/kg body weight in 95% ethanol, 5 days per week for 105 weeks. Survival of all dosed groups was similar to that of the vehicle controls. Mean body weights of dosed groups were within 10% of those of the vehicle control groups throughout the study. In the skin at the site of application, there were increased incidences of epidermal hyperplasia and hyperkeratosis in males and females administered 30 or 90 mg/kg. 2-YEAR DERMAL STUDY IN MICE: Groups of 50 male and 50 female mice were dermally administered 0, 10, 30, or 90 mg methyl trans-styryl ketone/kg body weight in 95% ethanol, 5 days per week for 105 weeks. Survival of all dosed groups was similar to that of the vehicle controls. Mean body weights of dosed groups were within 10% of those of the vehicle control groups throughout the study. In the skin at the site of application in males and females, there were treatment-related increased incidences of epidermal hyperplasia, hyperkeratosis, chronic inflammation, and melanocyte hyperplasia. Methyl trans-styryl ketone was mutagenic in Salmonella typhimurium strain TA100 when testing was conducted in the presence of rat liver microsomes (S9). No mutagenic activity was seen with methyl trans-styryl ketone in strain TA98 with or without S9 or in Escherichia coli strain WP2 uvrA/pKM101 in the absence of S9. With S9, inconsistent responses were seen in the E. coli tester strain. No increases in the frequencies of micronucleated normochromatic erythrocytes were seen in peripheral blood samples from male or female mice administered methyl trans-styryl ketone for 3 months via dosed feed or dermal application. Under the conditions of these 2-year dermal studies, there was no evidence of carcinogenic activity of methyl trans-styryl ketone in male or female F344/N rats or in male or female B6C3F1 mice administered 10, 30, or 90 mg/kg. Administration of methyl trans-styryl ketone resulted in nonneoplastic lesions of the skin at the site of application in male and female rats and mice. Synonyms: Acetocinnamone; benzalacetone; benzylideneacetone; methyl 2-phenylvinyl ketone; methyl styryl ketone; methyl β-styryl ketone; MSK; 4-phenyl-3-butene-2-one; 4-phenylbutenone; 2-phenylvinyl methyl ketone; styryl methyl ketone Systematic name: (3E)-4-Phenylbut-3-en-2-one.


Investigating the relationship between genome structure, composition, and ecology in prokaryotes.
Our thesis is that the DNA composition and structure of genomes are selected in part by mutation bias (GC pressure) and in part by ecology. To illustrate this point, we compare and contrast the oligonucleotide composition and the mosaic structure in 36 complete genomes and in 27 long genomic sequences from archaea and eubacteria. We report the following findings (1) High-GC-content genomes show a large underrepresentation of short distances between G(n) and C(n) homopolymers with respect to distances between A(n) and T(n) homopolymers; we discuss selection versus mutation bias hypotheses. (2) The oligonucleotide compositions of the genomes of Neisseria (meningitidis and gonorrhoea), Helicobacter pylori and Rhodobacter capsulatus are more biased than the other sequenced genomes. (3) The genomes of free-living species or nonchronic pathogens show more mosaic-like structure than genomes of chronic pathogens or intracellular symbionts. (4) Genome mosaicity of intracellular parasites has a maximum corresponding to the average gene length; in the genomes of free-living and nonchronic pathogens the maximum occurs at larger length scales. This suggests that free-living species can incorporate large pieces of DNA from the environment, whereas for intracellular parasites there are recombination events between homologous genes. We discuss the consequences in terms of evolution of genome size. (5) Intracellular symbionts and obligate pathogens show small, but not zero, amount of chromosome mosaicity, suggesting that recombination events occur in these species.


A comparative study of the grain-size distribution of surface dust and stormwater runoff quality on typical urban roads and roofs in Beijing, China.
The deposition of pollutants on impervious surfaces is a serious problem associated with rapid urbanization, which results in non-point-source pollution. Characterizing the build-up and wash-off processes of pollutants in urban catchments is essential for urban planners. In this paper, the spatial variation and particle-size distributions of five heavy metals and two nutrients in surface dust were analyzed, and the runoff water first-flush effect (FF30) and event-mean concentrations (EMCs) of 10 common constituents were characterized. The relationships between runoff variables and stormwater characteristics were examined from three typical urban impervious surfaces in Beijing, China. Dust on road surfaces with smaller grain sizes had higher pollutant concentrations, whereas concentrations of Mn, Zn, Fe, and TP in roof surface dust increased with grain size. Particles with grain sizes of 38-74 and 125-300 μm contributed most to the total pollutant load in roads, while particles with the smallest grain sizes (<38 μm) contributed most on roofs (23.46-41.71 %). Event-mean concentrations (EMCs) and FF30 values for most runoff pollutants tended to be higher on roofs than on roads. The maximum intensity (I max) and the antecedent dry days (ADD) were critical parameters for EMCs in roads, while ADD was the only dominant parameter for EMCs on our studied roof. The rainfall intensity (RI) and maximum intensity (I max) were found to be the parameters with the strongest correlation to the first-flush effect on both roads and roofs. Significant correlations of total suspended solids (TSS) concentration in runoff with grain-size fractions of surface dust indicated that coarser particles (74-300 μm) are most likely to contribute to the solid-phase pollutants, and finer particles (<38 μm) are likely the main source of dissolved pollutants.


Miniaturized spectrometers with a tunable van der Waals junction.
Miniaturized computational spectrometers, which can obtain incident spectra using a combination of device spectral responses and reconstruction algorithms, are essential for on-chip and implantable applications. Highly sensitive spectral measurement using a single detector allows the footprints of such spectrometers to be scaled down while achieving spectral resolution approaching that of benchtop systems. We report a high-performance computational spectrometer based on a single van der Waals junction with an electrically tunable transport-mediated spectral response. We achieve high peak wavelength accuracy (∼0.36 nanometers), high spectral resolution (∼3 nanometers), broad operation bandwidth (from ∼405 to 845 nanometers), and proof-of-concept spectral imaging. Our approach provides a route toward ultraminiaturization and offers unprecedented performance in accuracy, resolution, and operation bandwidth for single-detector computational spectrometers.


Learning the faculty role: using the evolving case story of professor able in an online master of nursing education program.
This article presents the use of a case story about a fictitious character, Professor Able, as a strategy to learn about the role of the nurse educator and to assist in the transition from clinical practice into that role. The story evolves over a 13-week semester in an engaging, asynchronous online environment where students explore what it means to be a nurse educator. The story of Professor Able provides insights into faculty issues such as academic freedom, integrity, governance, and diversity. Students' online discussions highlight the interactive learning experience and outcomes generated by the use of the case story. This teaching strategy offers support for nurses transitioning into the much-needed role of nurse educator.


Physicians in alcoholism: a study of current status and future needs.
Members of the American Medical Society on Alcoholism were studied to shed light on medical manpower needs in the alcoholism field. Findings were compared to AMA and federal survey data for all American physicians. AMSA respondents were broadly distributed among specialties: primarily psychiatrists, internists, and family practitioners. They were evenly divided between office-based and institutional practice, and most were on medical school faculties. Regarding alcoholism treatment, they generally agreed that Alcoholics Anonymous is necessary and that nonmedical counsellors are effective, although their attitudes on the etiology of alcoholism were divided. This sample of physicians in alcoholism, although not inclusive of all in the field, represents a diverse group, with differing clinical and educational needs.


[Steroid drugs and GM-CSF modulates activity of Egr-1 in glioma cells].
The Egr-1 protein is a transcriptional factor responsive to early growth. Transcriptional regulation of the promoter has been described like responsive to physical stress, osmotic changes, and cellular growth marker. However, there is no report about the pharmacological effect on the transcriptional regulation in gliomas. Hereby we report the modulation of the Egr-1 promoter transcriptional activity induced by the Granulocytes Macrophages Colony Stimulating Factor (GM-CSF) and steroid drugs in human glioma cells (CH235-GM Grade II, U373-GM Grade III, D54-GM Grade IV) using a reporter system transduced by a recombinant adenoviral vector AdEgr-1/luc7. Human glioma cells shows with different malignity grade (CH235-GM Grado II; U373-GM Grado III; D54-GM Grado IV) were transduced with no replicative adenoviral vector AdEgr-1/Luc7 and exposed to drugs as progesterone, β-estradiol and betametasone, and GM-CSF. Transcriptional activity of the egr-1 promoter was quantified by Luciferase reporter gene, cloned downstream to the tata box. Luciferase activity was quantified from whole cell proteins using luminometry assays. U373-GM cell line with GM-CSF, shows an increment on transcriptional activity of Egr-1 promoter, also in endogen way. U373-GM showed a positive regulation of Egr-1, with steroid drugs on the times analyzed. Steroid drugs as progesterone, β-estradiol and betametasone, shows a pleiotropic behavior on CH235-GM and D54-GM, glioma cell lines. Inhibition or activation response of Egr-1 promoter shows new framework to explore a mechanism of action of steroid drugs on genetic and epigenetic regulation on tumoral process.


Developmental abnormalities in supporting cell phalangeal processes and cytoskeleton in the Gjb2 knockdown mouse model.
Mutations in the GJB2 gene [which encodes connexin 26 (Cx26)] are the most common causes of hereditary hearing loss in humans, and previous studies showed postnatal development arrest of the organ of Corti in different Cx26-null mouse models. To explore the pathological changes and the mechanism behind the cochlear abnormalities in these mice further, we established transgenic mouse models by conditional knockdown of cochlear Cx26 at postnatal day (P) 0 and P8. Auditory brainstem responses were recorded and the morphological features in the organ of Corti were analyzed 18 days after Cx26 knockdown. Mice in the P0 knockdown group displayed severe hearing loss at all frequencies, whereas mice in the P8 knockdown group showed nearly normal hearing. In the P8 knockdown group, the organ of Corti displayed normal architecture, and no ultrastructural changes were observed. In the P0 knockdown group, the phalangeal processes of Deiter's cells did not develop into finger-like structures, and the formation of microtubules in the pillar cells was significantly reduced; moreover, the amount of acetylated α-tubulin was reduced in pillar cells. Our results indicate that Gjb2 participates in postnatal development of the cytoskeleton in pillar cells during structural maturation of the organ of Corti. In P0 knockdown mice, the reduction in microtubules in pillar cells might be responsible for the failure of the tunnel of Corti to open, and the malformed phalangeal processes might negatively affect the supporting framework of the organ of Corti, which would be a new mechanism of Gjb2-related hearing loss.


[CHANGING OF ISCHEMIC M. SOLEUS TETANIC CONTRACTION PARAMETERS IN RATS WITH CHRONIC ALCOHOL INTOXICATION].
This article deals with the changes of isolated ischemic m. soleus tetanus parameters in rats with chronic alcohol intoxication. The experiments were carried out on 15 male Wistar rats that were divided into three groups for 5 animals in each: group I (control) and two groups in which was induced hind limbs acute muscles ischemia: group II - rats without alcoholic intoxication, group III - rats with chronic alcoholic intoxication. Strain measurement muscle mechanical activity were conducted in isometric mode under conditions of direct electrical muscular preparation stimulation. It is proved that ischemic m. soleus tetanic force in rats with chronic alcoholic intoxication in comparison with rats without alcoholic intoxication does not significant changes. But signifycantly increases the reaching tetanus peak time. It is shown that in rats without alcoholic intoxication and with chronic alcoholic intoxication in comparison with intact animals, significantly decreases the duration of ischemic m. soleus stabile force level. It is shoved significant changes of individual muscles contraction time course of ischemic m. soleus tetanus in this rats group in comparison to intact animal. It is shown that these changes influence on successive muscular contraction efficiency of frequency summation in ischemic m. soleus tetanus and their speed-power characteristics.


Role of radiotherapy in adrenocortical carcinoma.
Adrenocortical carcinoma (ACC) is a rare and highly malignant tumor usually diagnosed in an advanced stage. Radiation therapy has been a poorly studied and underutilized therapeutic option. This retrospective analysis reviewed treatment courses for 14 patients with pathologically confirmed ACC treated between 1997 and 2012. Two patients were treated adjuvantly following surgery, and 12 were treated with palliative intent. Patients presented with stage II (n = 4), stage III (n = 7), and stage IV (n = 3) disease. Patients had a mean age of 51.5 years. Ten patients received chemotherapy before radiotherapy (RT), and 12 patients received surgery before RT, before receiving radiation at a mean of 17.8 months after diagnosis. In total, 20 sites were treated, 2 of which were in an adjuvant setting, and 18 of which were for palliative indications in 12 patients as follows: (1) pain/neuropathy (n = 10), (2) prophylactic treatment of asymptomatic recurrences (n = 3), and (3) prevention of imminent metastatic complications (n = 2), hemoptysis (n = 1), severe mass effect (n = 1), and brain metastasis (n = 1). Sites were treated to a median dose of 36.3 Gy (range, 17.5-60 Gy) in a median of 2.5 Gy/fraction (range, 1.8-4 Gy). At a mean follow-up of 22.0 months for the 2 patients given adjuvant RT, 1 patient did not have a local recurrence during a 14.3-month period of follow-up, and the other had a local recurrence 14.5 months after RT. At a mean follow-up of 11.3 months for the 12 patients receiving palliative RT, 10 patients had either a clinical or radiographic response. Of the courses of palliative RT that had adequate radiographic follow-up, 4 treatments (27%) resulted in a partial response. Eleven treatments (73%) that were able to be evaluated resulted in clinical improvement. Acute Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer toxicities observed in 7 patients included 3 grade 1, 4 grade 2, and 1 grade 3. No patient had acute toxicity of grade 4 or greater or any Radiation Therapy Oncology Group/European Organization for Research and Treatment of Cancer late toxicity of grade 1 or greater. This report is one of the largest to date examining the role of modern radiation techniques in the management of ACC. We conclude that radiation can be effective in the management of metastatic ACC, palliating local symptoms, and preventing complications from large metastases. Radiation should be considered as an option in multimodality management of ACC patients.


The role of amifostine on late normal tissue damage induced by pelvic radiotherapy with concomitant gemcitabine: an in vivo study.
In this in vivo study, we aimed to assess the radioprotective effect of amifostine on late normal tissue damage induced by gemcitabine concomitant with pelvic radiotherapy by histopathological and quantitative methods. Fifty-six male Wistar albino rats were randomly divided into seven experimental groups as follows: (I) gemcitabine, (II) radiation + gemcitabine, (III) radiation + gemcitabine + amifostine, (IV) radiation + amifostine, (V) sham radiation, (VI) amifostine, (VII) radiation. Irradiation was given to pelvic region with a dose of 25 Gy in 5 fractions. Amifostine was given for 30 min; gemcitabine was administered 24 h before the first fraction of radiotherapy. All animals were killed at the end of 4th month. Pathological examination was performed and the tissue collagen content was measured in bladder and rectal tissues. Fifty-one animals that were alive at the end of the follow-up period were analyzed. Thirty-five animals (68.6%) revealed grades I-III late effect in histopathological examination. We observed grade III colitis in 1 animal (radiation + gemcitabine) and bladder fibrosis in 4 animals (radiation and radiation + gemcitabine groups). There was no significant difference between any groups for bladder cystitis and fibrosis by Kruskal-Wallis method. Colitis was seen significantly lower in the radiation + gemcitabine + amifostine group (P = 0.0005). The collagen contents in the bladder and rectum of radiation and radiation + gemcitabine groups were markedly increased as compared to the sham group. This effect was reversed in the groups which received amifostine in addition to radiation and radiation + gemcitabine groups, but this difference was not significant. This study demonstrated that amifostine may have a beneficial effect in limiting rectal colitis from the radiosensitizing effect of gemcitabine.


Phosphorus limitation, soil-borne pathogens and the coexistence of plant species in hyperdiverse forests and shrublands.
Hyperdiverse forests occur in the lowland tropics, whereas the most species-rich shrublands are found in regions such as south-western Australia (kwongan) and South Africa (fynbos). Despite large differences, these ecosystems share an important characteristic: their soils are strongly weathered and phosphorus (P) is a key growth-limiting nutrient. Soil-borne pathogens are increasingly being recognized as drivers of plant diversity in lowland tropical rainforests, but have received little attention in species-rich shrublands. We suggest a trade-off in which the species most proficient at acquiring P have ephemeral roots that are particularly susceptible to soil-borne pathogens. This could equalize out the differences in competitive ability among co-occurring species in these ecosystems, thus contributing to coexistence. Moreover, effective protection against soil-borne pathogens by ectomycorrhizal (ECM) fungi might explain the occurrence of monodominant stands of ECM trees and shrubs amongst otherwise species-rich communities. We identify gaps in our knowledge which need to be filled in order to evaluate a possible link between P limitation, fine root traits, soil-borne pathogens and local plant species diversity. Such a link may help to explain how numerous plant species can coexist in hyperdiverse rainforests and shrublands, and, conversely, how monodominant stands can develop in these ecosystems.


Formal recycling of e-waste leads to increased exposure to toxic metals: an occupational exposure study from Sweden.
Electrical and electronic waste (e-waste) contains multiple toxic metals. However, there is currently a lack of exposure data for metals on workers in formal recycling plants. The objective of this study was to evaluate workers' exposure to metals, using biomarkers of exposure in combination with monitoring of personal air exposure. We assessed exposure to 20 potentially toxic metals among 55 recycling workers and 10 office workers at three formal e-waste recycling plants in Sweden. Workers at two of the plants were followed-up after 6 months. We collected the inhalable fraction and OFC (37-mm) fraction of particles, using personal samplers, as well as spot samples of blood and urine. We measured metal concentrations in whole blood, plasma, urine, and air filters using inductively coupled plasma-mass spectrometry following acid digestion. The air sampling indicated greater airborne exposure, 10 to 30 times higher, to most metals among the recycling workers handling e-waste than among the office workers. The exposure biomarkers showed significantly higher concentrations of chromium, cobalt, indium, lead, and mercury in blood, urine, and/or plasma of the recycling workers, compared with the office workers. Concentrations of antimony, indium, lead, mercury, and vanadium showed close to linear associations between the inhalable particle fraction and blood, plasma, or urine. In conclusion, our study of formal e-waste recycling shows that workers performing recycling tasks are exposed to multiple toxic metals.


Quality Assessment in Dobutamine Stress Echocardiography: What are the Clinical Predictors Associated With a Non-Diagnostic Test?
Non-diagnostic dobutamine stress echocardiography (ndDSE, failure to achieve 85% of maximal predicted heart rate (HR) without evidence of inducible ischemia) is an important limitation affecting quality of DSE testing. The objectives of this study were to identify the clinical variables associated with a non-diagnostic Dobutamine Stress Echocardiogram (ndDSE) and further evaluate the patterns of subsequent testing for myocardial ischemia. Consecutive DSE's over a 17 month period (January 2008 to June 2009) were studied. Baseline demographics, medical history, and vital signs were collected. Subsequent testing was determined for up to 6 months after the initial DSE. Univariate and multivariate logistic regression analysis was performed to identify clinical factors associated with ndDSE. Of 467 total DSE, 314 (67%) were negative for ischemia, 69 (15%) were positive, and 84 (18%) were ndDSE. Of those recommended for further nuclear MPI testing 12 (14%) had an ndDSE compared to 16 (4%) patients with a diagnostic DSE (P = 0.001). Fifty percent of the ndDSE nuclear MPI tests were positive for ischemia. In the univariate analysis, Diabetes Mellitus (DM; P = 0.003), calcium channel antagonist (CCA) use (P = 0.047), Hypertension (HTN; P = 0.06), low baseline HR (P < 0.001), and younger age group (P = 0.02) were predictive of ndDSE. Of these, all except CCA use remained independent predictors of ndDSE in multivariate analysis. A 4 variable model for predicting ndDSE was developed from the multivariate logistic regression displayed in Table 1 (age and baseline HR were categorized and scored 0-2; DM and HTN were scored as 0 (absent) or 1 (present)). Figure 2 demonstrates how risk of ndDSE correlated with a higher score, with each increment having an odds ratio of 2.1 (P < 0.001). DM, HTN, younger age, and lower baseline HR affect the quality of DSE testing, resulting in non-diagnostic tests. A model combining these factors can identify patients most likely to have this outcome. Identification of this cohort may improve referral patterns and improve the quality of stress testing.


SOD therapeutics: latest insights into their structure-activity relationships and impact on the cellular redox-based signaling pathways.
Superoxide dismutase (SOD) enzymes are indispensable and ubiquitous antioxidant defenses maintaining the steady-state levels of O2·(-); no wonder, thus, that their mimics are remarkably efficacious in essentially any animal model of oxidative stress injuries thus far explored. Structure-activity relationship (half-wave reduction potential [E1/2] versus log kcat), originally reported for Mn porphyrins (MnPs), is valid for any other class of SOD mimics, as it is dominated by the superoxide reduction and oxidation potential. The biocompatible E1/2 of ∼+300 mV versus normal hydrogen electrode (NHE) allows powerful SOD mimics as mild oxidants and antioxidants (alike O2·(-)) to readily traffic electrons among reactive species and signaling proteins, serving as fine mediators of redox-based signaling pathways. Based on similar thermodynamics, both SOD enzymes and their mimics undergo similar reactions, however, due to vastly different sterics, with different rate constants. Although log kcat(O2·(-)) is a good measure of therapeutic potential of SOD mimics, discussions of their in vivo mechanisms of actions remain mostly of speculative character. Most recently, the therapeutic and mechanistic relevance of oxidation of ascorbate and glutathionylation and oxidation of protein thiols by MnP-based SOD mimics and subsequent inactivation of nuclear factor κB has been substantiated in rescuing normal and killing cancer cells. Interaction of MnPs with thiols seems to be, at least in part, involved in up-regulation of endogenous antioxidative defenses, leading to the healing of diseased cells. Mechanistic explorations of single and combined therapeutic strategies, along with studies of bioavailability and translational aspects, will comprise future work in optimizing redox-active drugs.


Reductive activation of nitrate reductases.
Protein film voltammetry of Paracoccus pantotrophus respiratory nitrate reductase (NarGH) and Synechococcus elongatus assimilatory nitrate reductase (NarB) shows that reductive activation of these enzymes may be required before steady state catalysis is observed. For NarGH complementary spectroscopic studies suggest a structural context for the activation. Catalytic protein film voltammetry at a range of temperatures has allowed quantitation of the activation energies for nitrate reduction. For NarGH with an operating potential of ca. 0.05 V the activation energy of ca. 35 kJ mol-1 is over twice that measured for NarB whose operating potential is ca. -0.35 V.


The complex I subunit NDUFA10 selectively rescues Drosophila pink1 mutants through a mechanism independent of mitophagy.
Mutations in PINK1, a mitochondrially targeted serine/threonine kinase, cause autosomal recessive Parkinson's disease (PD). Substantial evidence indicates that PINK1 acts with another PD gene, parkin, to regulate mitochondrial morphology and mitophagy. However, loss of PINK1 also causes complex I (CI) deficiency, and has recently been suggested to regulate CI through phosphorylation of NDUFA10/ND42 subunit. To further explore the mechanisms by which PINK1 and Parkin influence mitochondrial integrity, we conducted a screen in Drosophila cells for genes that either phenocopy or suppress mitochondrial hyperfusion caused by pink1 RNAi. Among the genes recovered from this screen was ND42. In Drosophila pink1 mutants, transgenic overexpression of ND42 or its co-chaperone sicily was sufficient to restore CI activity and partially rescue several phenotypes including flight and climbing deficits and mitochondrial disruption in flight muscles. Here, the restoration of CI activity and partial rescue of locomotion does not appear to have a specific requirement for phosphorylation of ND42 at Ser-250. In contrast to pink1 mutants, overexpression of ND42 or sicily failed to rescue any Drosophila parkin mutant phenotypes. We also find that knockdown of the human homologue, NDUFA10, only minimally affecting CCCP-induced mitophagy, and overexpression of NDUFA10 fails to restore Parkin mitochondrial-translocation upon PINK1 loss. These results indicate that the in vivo rescue is due to restoring CI activity rather than promoting mitophagy. Our findings support the emerging view that PINK1 plays a role in regulating CI activity separate from its role with Parkin in mitophagy.


Employee and customer injury during violent crimes in retail and service businesses.
We sought to compare the frequency and risk factors for employees and customers injured during crimes in retail (convenience, grocery, and liquor stores) and service businesses (bars, restaurants, motels). A total of 827 retail and service businesses in Los Angeles were randomly selected. Police crime reports (n=2029) from violent crimes that occurred in these businesses from January 1996 through June 2001 were individually reviewed to determine whether a customer or an employee was injured and to collect study variables. A customer injury was 31% more likely (95% confidence interval [CI]=1.11, 1.51) than an employee injury during a violent crime. Customer injury was more frequent than employee injury during violent crimes in bars, restaurants, convenience stores, and motels but less likely in grocery or liquor stores. Injury risk was increased for both employees and customers when resisting the perpetrator and when the perpetrator was suspected of using alcohol. Customers had an increased risk for injury during crimes that occurred outside (relative risk [RR]=2.01; 95% CI=1.57, 2.58) and at night (RR=1.79; 95% CI=1.40, 2.29). Security programs should be designed to protect customers as well as employees.


[Treatment of periodontitis in cases of diabetes mellitus is always worthwhile!].
In the May edition of The Nederlands Tijdschrift voor Tandheelkunde the editorial commentary was devoted to the question to what extent the treatment of periodontitis in patients with diabetes mellitus is worthwhile. B.G. Loos, W.J. Teeuw, V. Gerdes and F. Abbas offer a response to this commentary and argue that the treatment of periodontitis in patients with diabetes mellitus is always worthwhile.


[Immunohistochemical study of the distribution of collagens (type I, III, IV), fibronectin, factor XIIIa and factor VIII related antigen in alcoholic liver disease].
Histopathological changes in alcoholic liver disease are characterized by Mallory body, fatty liver, and peculiar fibrosis such as pericentral fibrosis, pericellular fibrosis, stellate fibrosis originating from Glisson's sheath, and swelling of the liver cells. However, the mechanism of fibrosis remains unknown. The author tried to demonstrate, by the immunohistochemical method, fibronectin (FN), collagen type I, III, IV (IC, IIIC, IVC), factor XIIIa, and factor VIII related antigen (VIII RAg), all of which are related to the process of wound healing, in order to clarify the mechanism of fibrosis in alcoholic liver disease. With the progress of fibrosis, FN was demonstrated in the sinusoidal wall, as well as the portal areas and around the central veins. IC and IIIC were positive in the fibrotic area extending into the lobules and pericellular fibrosis. IVC was similar to FN and was positive in the sinusoidal wall. Scattered XIIIa positivity was recognized in the fibrotic areas, especially in the foci of active fibrosis. VIII RAg was positive in the capillary vessels invading the lobules through the limiting plates. The distribution of these proteins in the fibrotic area in alcoholic liver disease closely resembled that in the wound healing process. It is concluded that fibrosis occurring in the periportal area is essentially the same as in the wound healing process and the mechanism of fibrosis in alcoholic liver disease is thought to be active fibroplasia.


Hepatitis B virus HBx protein interactions with the ubiquitin proteasome system.
The hepatitis B virus (HBV) causes acute and chronic hepatitis, and the latter is a major risk factor for the development of hepatocellular carcinoma (HCC). HBV encodes a 17-kDa regulatory protein, HBx, which is required for virus replication. Although the precise contribution(s) of HBx to virus replication is unknown, many viruses target cellular pathways to create an environment favorable for virus replication. The ubiquitin proteasome system (UPS) is a major conserved cellular pathway that controls several critical processes in the cell by regulating the levels of proteins involved in cell cycle, DNA repair, innate immunity, and other processes. We summarize here the interactions of HBx with components of the UPS, including the CUL4 adaptor DDB1, the cullin regulatory complex CSN, and the 26S proteasome. Understanding how these protein interactions benefit virus replication remains a challenge due to limited models in which to study HBV replication. However, studies from other viral systems that similarly target the UPS provide insight into possible strategies used by HBV.


Mutations of the immunoglobulin heavy chain variable region gene in CD99-deficient BJAB cell line.
Hodgkin's disease (HD) is a lymphoid neoplasm characterized by a low frequency of malignant giant tumor cells, known as Hodgkin's and Reed-Sternberg (HRS) cells. Sequence analysis of the immunoglobulin heavy chain hypervariable region (IgH V) genes of HRS cells revealed multiple nucleotide substitutions, indicating somatic mutations, and suggested that HRS cells originate from germinal center B cells or their progeny. We previously reported that CD99-antisense transfected B cell lines led to the generation of cells with a HRS phenotype. Because it is considered that HRS cells in HD carry somatic mutations of the IgH genes, we assume that somatic mutation may take place in the IgH genes of HRS-like cells which do not express CD99. Here we report that CD99 downregulated BJAB cell line has several mutations in IgH V genes. The frequency of mutation was 5.2 x 10(-4) mut.bp(-1) out of total sequenced cell clones. On the contrary, control vector transfected BJAB cell line or CD99 downregulated IM9 cell line did not show any mutations on single strand conformational polymorphism (SSCP) and sequence analysis. We expect that the analysis of the mutation pattern of the CD99-deficient BJAB cell line might be the basis for the understanding of the molecular and cellular mechanism that regulate somatic mutation and B cell selection.


Differential influence of nutrient-starved Mycobacterium tuberculosis on adaptive immunity results in progressive tuberculosis disease and pathology.
When infected with Mycobacterium tuberculosis, most individuals will remain clinically healthy but latently infected. Latent infection has been proposed to partially involve M. tuberculosis in a nonreplicating stage, which therefore represents an M. tuberculosis phenotype that the immune system most likely will encounter during latency. It is therefore relevant to examine how this particular nonreplicating form of M. tuberculosis interacts with the host immune system. To study this, we first induced a state of nonreplication through prolonged nutrient starvation of M. tuberculosis in vitro. This resulted in nonreplicating persistence even after prolonged culture in phosphate-buffered saline. Infection with either exponentially growing M. tuberculosis or nutrient-starved M. tuberculosis resulted in similar lung CFU levels in the first phase of the infection. However, between week 3 and 6 postinfection, there was a very pronounced increase in bacterial levels and associated lung pathology in nutrient-starved-M. tuberculosis-infected mice. This was associated with a shift from CD4 T cells that coexpressed gamma interferon (IFN-γ) and tumor necrosis factor alpha (TNF-α) or IFN-γ, TNF-α, and interleukin-2 to T cells that only expressed IFN-γ. Thus, nonreplicating M. tuberculosis induced through nutrient starvation promotes a bacterial form that is genetically identical to exponentially growing M. tuberculosis yet characterized by a differential impact on the immune system that may be involved in undermining host antimycobacterial immunity and facilitate increased pathology and transmission.


Diffusion Tensor Imaging of Visual Pathway Abnormalities in Five Glaucoma Animal Models.
To characterize the visual pathway integrity of five glaucoma animal models using diffusion tensor imaging (DTI). Two experimentally induced and three genetically determined models of glaucoma were evaluated. For inducible models, chronic IOP elevation was achieved via intracameral injection of microbeads or laser photocoagulation of the trabecular meshwork in adult rodent eyes. For genetic models, the DBA/2J mouse model of pigmentary glaucoma, the LTBP2 mutant feline model of congenital glaucoma, and the transgenic TBK1 mouse model of normotensive glaucoma were compared with their respective genetically matched healthy controls. DTI parameters, including fractional anisotropy, axial diffusivity, and radial diffusivity, were evaluated along the optic nerve and optic tract. Significantly elevated IOP relative to controls was observed in each animal model except for the transgenic TBK1 mice. Significantly lower fractional anisotropy and higher radial diffusivity were observed along the visual pathways of the microbead- and laser-induced rodent models, the DBA/2J mice, and the LTBP2-mutant cats compared with their respective healthy controls. The DBA/2J mice also exhibited lower axial diffusivity, which was not observed in the other models examined. No apparent DTI change was observed in the transgenic TBK1 mice compared with controls. Chronic IOP elevation was accompanied by decreased fractional anisotropy and increased radial diffusivity along the optic nerve or optic tract, suggestive of disrupted microstructural integrity in both inducible and genetic glaucoma animal models. The effects on axial diffusivity differed between models, indicating that this DTI metric may represent different aspects of pathological changes over time and with severity.


HER2 signaling regulates HER2 localization and membrane retention.
ErbB2/HER2/Neu is a receptor tyrosine kinase that is overexpressed in 25-30% of human breast cancers, usually associated with amplification of the ERBB2 gene. HER2 has no recognized ligands and heterodimers between HER2 and EGFR (ErbB1/HER1) or HER2 and ErbB3/HER3 are important in breast cancer. Unlike other ErbB family members, HER2 is resistant to internalization and degradation, and remains at the cell surface to signal for prolonged periods after it is activated. Although the mechanisms underlying retention of HER2 at the cell surface are not fully understood, prior studies have shown that, in order to avoid internalization, HER2 must interact with the chaperone, HSP90, and the calcium pump, PMCA2, within specific plasma membrane domains that protrude from the cell surface. In this report, we demonstrate that HER2 signaling, itself, is important for the formation and maintenance of membrane protrusions, at least in part, by maintaining PMCA2 expression and preventing increased intracellular calcium concentrations. Partial genetic knockdown of HER2 expression or pharmacologic inhibition of HER2 signaling causes the depletion of membrane protrusions and disruption of the interactions between HER2 and HSP90. This is associated with the ubiquitination of HER2, its internalization with EGFR or HER3, and its degradation. These results suggest a model by which some threshold of HER2 signaling is required for the formation and/or maintenance of multi-protein signaling complexes that reinforce and prolong HER2/EGFR or HER2/HER3 signaling by inhibiting HER2 ubiquitination and internalization.


Cultural Differences in Susceptibility to Stereotype Threat: France versus India.
Negative aging stereotypes make older adults perform below their true potential in a number of cognitive domains. This phenomenon, known as Age-Based Stereotype Threat, is currently viewed as a powerful factor contributing to an overestimation of cognitive decline in normal aging. However, age-based stereotype threat has been investigated almost exclusively in Western countries. Whether this phenomenon is universal or culture-specific is unknown. Here, we first ran a pilot study (N = 106) in which we assessed French and Indian participants' attitudes towards aging. Then, we assessed stereotype threat effects on arithmetic problem-solving performance and strategies in French and Indian older adults (N = 104). We found that French older adults have more negative implicit attitudes towards aging than Indian older adults. We also found that culture modulates age-based stereotype threat effects. Whereas French older adults experienced stereotype threat on both selection and execution of strategies on all arithmetic problems, Indian older adults experienced this threat only in their strategy selection on harder problems. Most interestingly, cultural differences emerged on arithmetic problems under stereotype threat condition, where otherwise no cultural differences were found in the control condition. Our findings have important implications for understanding how cultural contexts change aging effects on human cognition and age-related difference in cognitive performance.


Ecophysiological characterization of ammonia-oxidizing archaea and bacteria from freshwater.
Aerobic biological ammonia oxidation is carried out by two groups of microorganisms, ammonia-oxidizing bacteria (AOB) and the recently discovered ammonia-oxidizing archaea (AOA). Here we present a study using cultivation-based methods to investigate the differences in growth of three AOA cultures and one AOB culture enriched from freshwater environments. The strain in the enriched AOA culture belong to thaumarchaeal group I.1a, with the strain in one enrichment culture having the highest identity with "Candidatus Nitrosoarchaeum koreensis" and the strains in the other two representing a new genus of AOA. The AOB strain in the enrichment culture was also obtained from freshwater and had the highest identity to AOB from the Nitrosomonas oligotropha group (Nitrosomonas cluster 6a). We investigated the influence of ammonium, oxygen, pH, and light on the growth of AOA and AOB. The growth rates of the AOB increased with increasing ammonium concentrations, while the growth rates of the AOA decreased slightly. Increasing oxygen concentrations led to an increase in the growth rate of the AOB, while the growth rates of AOA were almost oxygen insensitive. Light exposure (white and blue wavelengths) inhibited the growth of AOA completely, and the AOA did not recover when transferred to the dark. AOB were also inhibited by blue light; however, growth recovered immediately after transfer to the dark. Our results show that the tested AOB have a competitive advantage over the tested AOA under most conditions investigated. Further experiments will elucidate the niches of AOA and AOB in more detail.


Real-Time In Vivo Hepatotoxicity Monitoring through Chromophore-Conjugated Photon-Upconverting Nanoprobes.
Drug toxicity is a long-standing concern of modern medicine. A typical anti-pain/fever drug paracetamol often causes hepatotoxicity due to peroxynitrite ONOO- . Conventional blood tests fail to offer real-time unambiguous visualization of such hepatotoxicity in vivo. Here we report a luminescent approach to evaluate acute hepatotoxicity in vivo by chromophore-conjugated upconversion nanoparticles. Upon injection, these nanoprobes mainly accumulate in the liver and the luminescence of nanoparticles remains suppressed owing to energy transfer to the chromophore. ONOO- can readily bleach the chromophore and thus recover the luminescence, the presence of ONOO- in the liver leads to fast restoring of the near-infrared emission. Taking advantages of the high tissue-penetration capability of near-infrared excitation/emission, these nanoprobes achieve real-time monitoring of hepatotoxicity in living animals, thereby providing a convenient screening strategy for assessing hepatotoxicity of synthetic drugs.


[The intervention of reduced glutathione in neutrophil activation in the process of extracorporeal circulation].
To study the intervention of reduced glutathione for neutrophil activation in the process of extracorporeal circulation. 20 congenital heart disease children aged from 6 to 12 years were selected randomly, divided into control group and interventional group, patients of interventional group were added reduced glutathione (30 mg/kg) in priming fluid, then phlebotomized on the time of 24 hours before operation, 20 minutes after CPB, rewarming, the end of CPB, 4 hours after operation, 24 hours after operation. After separated neutrophil, we distilled nuclear protein and mensurated the activation of nuclear factor-kappaB by EMSA, assayed the expression of ICAM-1. by immunohistochemistry. Neutrophil nuclear factor-kappaB has been activated on the time of 20 minutes after CPB, arrives at the top on the time of rewarming, and resumes 24 hours after operation; ICAM-1 expresses on the time of rewarming, arrives at the top 4 hours after operation in control group. The data of interventional group is lower than the control group. Neutrophil nuclear factor-kappaB is activated and ICAM-1 is expressed in the process of extracorporeal circulation and play an important role in the inflammatory response happening in extracorporeal circulation, which could be relieved by reduced glutathione.


Designing experiments that aid in the identification of regulatory networks.
Predictive mathematical models of the interactions of a genetic network can provide insight into the mechanisms of gene regulation, the role of various genes within a network and how multiple genes interact leading to complex traits. However, identification of the parameters and interactions is currently a limiting step in the development of such models. This work reviews the state of the art for design of experiments in biological systems and demonstrates the need for improved design of experiments through the use of a model system. Appropriate design of experiments has a profound impact on the ability to identify a model and on the quality of resulting identified model. Key issues include the selection of appropriate input sequences (e.g. random, independent multivariate inputs) and the selection of the sampling frequencies. This work demonstrates that these issues are especially important in the identification of biochemical networks and that the traditional biochemical approach is incapable of truly identifying the behavior present in such networks.


PDE4 inhibition prevents preterm delivery induced by an intrauterine inflammation.
The aim of this study was to explore the anti-inflammatory properties of phosphodiesterase-4 (PDE4) inhibitors in vivo and their potential ability to prevent inflammation-induced preterm delivery. Indeed, intrauterine inflammation is the major etiology of very preterm delivery, the leading cause of neonatal mortality and morbidity. Intrauterine injection of Escherichia coli LPS in 15-day-pregnant mice induced an increase of PDE4 activity and PDE4B expression at the maternofetal interface, a rise of amniotic fluid levels of TNF-alpha, IL-1beta, IL-6, and IL-10 and provoked massive preterm delivery and fetal demise. Selective PDE4 inhibition by rolipram prevented the rise in the proinflammatory cytokines. Following the nuclear translocation of the transcription factor NFkappaB, as a marker of cellular activation after the inflammatory challenge, showed a time-dependent sequential activation of the gestational tissues, from the uterine mesometrial to the fetal compartment, particularly in the glycogen-trophoblastic cells of the placenta. This activation was disrupted by PDE4 inhibition, and inflammation-induced preterm delivery and fetal demise were prevented. PDE4 selective inhibitors may thus represent a novel effective treatment to delay inflammation-induced preterm delivery and to prevent adverse outcomes in infants.


Glucocorticoids confer normal serum/growth factor-dependent growth regulation to Fu5 rat hepatoma cells in vitro. Sequential expression of cell cycle-regulated genes without changes in intracellular calcium or pH.
Glucocorticoid hormones induced a stringent dependence on serum for the in vitro proliferation of Fu5 rat hepatoma cells by suppressing the growth rate and final quiescent cell density. Treatment of dexamethasone-suppressed quiescent Fu5 with serum plus insulin caused a rapid reinitiation of cellular proliferation and DNA synthesis that peaked at 16 h. RNA dot blot analysis of this time course showed that the transcript levels for the proto-oncogenes c-fos, c-myc, and c-rasKi peaked at 0.5, 2, and 4 h, respectively, while expression of c-rasHa and ornithine decarboxylase transcripts rose steadily during 16 h. Microspectrofluorimetric measurements of cytosolic calcium (Ca2+i) with fura-2 showed that insulin and serum, alone or in combination, elicited no changes in Ca2+i over a 50-min time course, although ATP, which is not a mitogen, induced large increases in Ca2+i. Cytosolic pH, pHi, was also measured using 2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein. Insulin and serum, alone or in combination, did not cause pHi to increase in either 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) (pHi 7.17)- or HCO3/CO2 (pHi 7.19)- buffered media. Acid-loading of cells with NH4Cl indicated that both quiescent and proliferating Fu5 cells have equally active, amiloride-sensitive Na/H exchangers. Therefore, induction of DNA synthesis and proto-oncogene expression occurs in Fu5 epithelial tumor cells in the absence of any short term increases of pHi or Ca2+i.


[Changes in the volutin levels in yeast cells during cell cycle].
Dimeric distribution of the cell number of Saccharomyces cerevisiae is analysed according to the volume and content of volutin in them. Dynamics of the age composition of the population in time when changing the medium limiting growth for the balanced one is considered. The data obtained indicate that volutin may serve as a regulator when the yeast cells pass the starting point in the division cycle, however, this event has no contrary effect on volutin accumulation.


Military surgery at the turn of the century.
The authors present a summary of current postgraduate education status in the field of military clinical specializations with the focus on surgery and military surgery. Recent organizational changes in medical care within the Czech Army, along with changes associated with the new legal directives regulating the acquirement of professional and specialized medical qualification, will require a new approach to the training of surgeons and other physicians in the Czech Republic. This will involve new educational goals, adaptation of specialized curricula and introduction of systematic quality measurement methods of teaching and training.


Essential tremor: diagnosis and treatment.
Essential tremor is a common movement disorder in adults that interferes with the performance of functional and social activities. Differentiation of essential tremor from other tremor syndromes is important in order to provide appropriate patient education and therapy. The mainstays of pharmacotherapy are propranolol and primidone; however, in selected patients, agents such as alcohol, benzodiazepines, botulinum toxin, and gabapentin may provide symptomatic benefits. Advances in surgical interventions, such as stereotactic thalamotomy and thalamic deep brain stimulation, offer patients an alternative treatment modality when pharmacotherapy is inadequate. A treatment algorithm is provided to guide clinicians in the management of patients with essential tremor.


Beetroot-Carrot Juice Intake either Alone or in Combination with Antileukemic Drug 'Chlorambucil' As A Potential Treatment for Chronic Lymphocytic Leukemia.
Chronic lymphocytic leukemia (CLL) is one of the chronic lymphoproliferative disorders (lymphoid neoplasms). It is characterized by a progressive accumulation of functionally incompetent lymphocytes. Patients with leukemia often seek unconventional treatments not prescribed by hematologist in order to improve their cancer treatment outcome or to manage symptoms. In the present report, a 76-year-old patient was diagnosed with B-cell chronic lymphocytic leukemia (B-CLL). Beetroot-carrot juice is used as a complementary and or/alternative therapy used in conjunction with conventional leukemic treatment (chlorambucil) that has been a standard first-line chemotherapeutic agent for patients with CLL and known to have serious and undesirable side-effects. After one month and 15 days of administration of beetroot-carrot juice therapy, the patient had improved appetite, a sense of general well-being and increased vigor daily activities. Furthermore, beetroot-carrot juice was used as an adjuvant to chlorambucil resulted in a substantial reduction in leukocytes and lymphocytes count in peripheral blood and improvement in the relevant biochemical parameters. Beetroot-carrot juice can be used as an effective treatment for CLL alone or in combination with chlorambucil when taken orally with regular diet on daily basis.


The Role of Serum Albumin Level during Hospitalization as a Predictor of Complications and Mortality in Children with Burns.
Serum albumin can function as a potential biomarker to determine the severity of the injury and clinical staging of children with burns. Therefore, in this study, we investigated the association between serum albumin level and complications and mortality rate in children with burns. In this descriptive-analytic cross-sectional study, 85 patients younger than 18 years with burns who were admitted to Shahid Motahari hospital between 2021 and 2022 were studied. Demographic information, including patients' age, sex, weight, underlying diseases, medical information, albumin level, and C-reactive protein (CRP), was obtained from patient records. Patients were observed until discharge. The independent t-test, chi-square, Pearson correlation, and logistic regression were used for analysis and to examine the predictive role of albumin. Out of 85 patients, 47 and 38 were boys and girls, respectively. The mean age of the participants was 3.69 ± 3.09 years. The mean length of hospital stay was 2.3 days, with a median of 1.5 days. The mean percentage of burns was 23.44 ± 16.50, and burn grade 2 was the most common. A total of 25 patients (29.41%) were admitted to the intensive care unit (ICU), and 13 deaths (15.29%) were observed among the patients. The mean albumin level was significantly lower than in other patients with outcomes of pulmonary infection, sepsis, renal failure, ICU admission, and death (P < 0.001). Serum Albumin has a significant predictive value in death, pulmonary infection, sepsis, admission to the ICU, and renal failure. Serum albumin may be a good prognostic marker associated with morbidity and mortality.


Pancreatic surgery in Italy. Criteria to identify the hospital units and the tertiary referral centers entitled to perform it.
Indicators of effectiveness and quality of care are urgently needed to improve the surgical outcomes. This is particularly felt in the field of complex surgical fields, such as the HPB one. International and national studies have documented an association between the large number of pancreatic surgical procedures and the outcome quality. The aim of this paper is to suggest reliable structural requirements and surgical volume to support pancreatic surgical accreditation, preserving patient's safety. Moreover, an accreditation program is outlined.


Parasite development and visceral pathology in Galba truncatula co-infected with Fasciola hepatica and Paramphistomum daubneyi.
Histological investigations in Galba truncatula naturally or experimentally co-infected with Fasciola hepatica and Paramphistomum daubneyi were carried out to study parasite development and the responses of the digestive gland and kidney of snails, as larval forms of these digeneans often use these two sites for their growth within the snail's body. The number of live rediae per snail ranged from 2.4 to 4.2 for the dominating parasite (it developed in the digestive gland) and was less than 2.0 for the other species. When the dominating species was F. hepatica, most snails harboured cercariae-containing rediae; if this parasite was P. daubneyi, procercariae-containing rediae with or without free procercariae were observed in most snails. In contrast, most rediae of the other species were immature. The pathology caused by the dominating species in the digestive gland was greater than that recorded in the kidney, where the other parasite was generally located. The most frequent tissue lesions in the digestive gland were generalized epithelial necrosis and epithelial reconstitution. In the kidney, multifocal epithelial necrosis was frequently observed, particularly when P. daubneyi was the dominating species. The frequencies of lesions in the digestive gland agreed with percentages reported by our team in other snails mono-infected with F. hepatica or P. daubneyi. In contrast, multifocal necrosis in the kidney was clearly greater in the present study and this finding might be explained by assuming that a sufficient number of free larvae within the snail would be necessary for the development of epithelial necrosis in the whole kidney.


Ultrasonography to assist with timing of spawning in European eel.
Ovulation in the European eel can be induced by injection of DHP (17α,20β-dihydroxy-4-pregnen-3-one). The timing of injection is based on the developmental stage of the oocytes. The oocyte stage is determined by invasive biopsy or through external indicators of the oocyte hydration response: body weight index (BWI) and body girth index (BGI). However, in European eel, BWI and BGI are inaccurate indicators because the individual hydration response is highly variable. The aim of this study was to identify indicators of oocyte maturation non-invasively by applying ultrasonography. Ultrasound parameters that were determined were the body wall thickness; diameter of the cloaca opening; grey scale median (GSM) of the ovary, and diameters (vertical and horizontal) and surface area of ovary, liver and swim bladder. Data of farmed and wild eels were combined for correlation analyses and principal component analyses (PCA). Ovary and liver parameters were not correlated with the average oocyte stage. Body wall thickness was negatively correlated to the average oocyte stage but variability was high. The swim bladder vertical diameter (Sbv) was negatively correlated with the average oocyte stage and with the oocyte diameter. Swim bladder area (Sba) was also negatively correlated to the average oocyte stage but less accurate. Results showed that during the final stages of oocyte maturation, the swim bladder becomes smaller. This is most probably due to the pressure that is caused by the increasing ovaries as a result of the oocyte hydration response. Females with an average oocyte stage between 4.5 and 7.0 and an average oocyte diameter higher than 800 μm, had Sbv values decreasing from 4.3 to 2.4 mm and Sba values from 52 down to 11 mm2. The correlation between Sbv and oocyte stage is more strict than for BWI or BGI which makes Sbv not only a parameter that can be determined non-invasively but also one that is more accurate in indicating the oocyte developmental stage. Therefore, ultrasonography, and in particular the ultrasound imaging of the swim bladder, represents a useful tool to assist with the timing of spawning as induced by injection of DHP.


Studies on species sensitivity to the dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. Part 1: Systemic administration.
Several parameters necessary for the expression of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced neurotoxicity to dopaminergic neurons were examined in both mice and rats in order to determine if differences in these processes might underlie the marked differences in the sensitivity of the two species to the neurotoxic effects of MPTP. Monoamine oxidase-B activity was greater in brain tissues from rats than from mice. The kinetics of 1-methyl-4-phenylpyridinium (MPP+) uptake into neostriatal synaptosomal preparations from the two species were similar. Brain and neostriatal levels of MPP+ were 2-fold higher in rats after the administration of MPTP at 60 mg/kg and were 10 to 20 times higher in rats than in mice after MPTP treatment which produced similar decrements in the content of neostriatal dopamine. MPP+ concentrations in the extracellular fluid of the neostriatum of the two species were similar after the administration of the same dose of MPTP (40 mg/kg). However, this dose induced a 40-fold increase in neostriatal dopamine efflux in mice, whereas in rats only a 3-fold increase was observed. In addition, pretreatment of rats with guanethidine, a ganglionic blocking agent, permitted the use of high doses of MPTP which resulted in substantial damage to the striatal dopaminergic nerve terminals. It is concluded that nigrostriatal dopaminergic neurons in the rat require exposure to a much higher concentration of MPP+ than do those in mice for the induction of toxicity.


An effective strategy for the monitoring of microplastics in complex aquatic matrices: Exploiting the potential of near infrared hyperspectral imaging (NIR-HSI).
Contamination by microplastics (MP) represents a critical environmental challenge with potential consequences at ecosystem, economic and societal levels. As the marine system is the final sink for MP, there is an urgent need to develop methods for the monitoring of synthetic particles in different marine compartments and sample matrices. Extensive evaluations are hindered by time and costs associated with to conventional MP spectroscopic analyses. The potential of near infrared hyperspectral imaging (NIR-HSI) has been recently evaluated. However, NIR-HSI has been poorly studied so far, limitedly to the detection of large particles (>300 μm), and its capability for direct characterization of MP in real marine matrices has not been considered yet. In the present study, a rapid near infrared hyperspectral imaging (NIR-HSI) method, coupled with a customised normalised difference image (NDI) strategy for data processing, is presented and used to detect MP down to 50 μm in environmental matrices. The proposed method is largely automated, without the need for extensive data processing, and enabled a successful identification of different polymers, both in surface water and mussel soft tissue samples, as well as on real field samples with environmentally occurring MP. NIR-HSI is applied directly on filters, without the need for particles pre-sorting or multiple sample purifications, avoiding time consuming procedures, airborne contaminations, particle degradation and loss. Thanks to the time and cost effectiveness, a large-scale implementation of this method would enable to extensively monitor the MP presence in natural environments for assessing the ecological risk related to MP contamination.


Mitochondrial stress and glycoxidation increase with decreased kidney function.
Mitochondrial stress increases the production of fumarate, an intermediate of the Krebs cycle. Fumarate non-enzymatically reacts with the thiol group of cysteine, leading to the production of S-(2-succinyl)cysteine. Here, we quantified the concentration of fumarate, the free form of S-(2-succinyl)cysteine, and advanced glycation end-products, including N ε-(carboxymethyl)lysine and N δ-(5-hydro-5-methyl-4-imidazolone-2-yl)-ornithine, in the serum of chronic kidney disease patients, using liquid chromatography-tandem mass spectrometry and an enzymatic assay. In a cross-sectional study, we evaluated the difference in metabolite concentration between healthy individuals (n = 22) and kidney transplant patients (n = 93). Additionally, we evaluated the metabolite concentration of end-stage renal disease patients (n = 17) before and 1, 3, 6, and 12 months after transplantation, in a longitudinal study. While the S-(2-succinyl)cysteine and AGEs levels were significantly increased in accordance with the rising chronic kidney disease severity, they were significantly decreased after transplantation. However, fumarate levels were only significantly different in end-stage renal disease patients. The S-(2-succinyl)cysteine levels correlated with the pre-existing kidney function marker. This study demonstrates that mitochondrial metabolic disorders contribute to impaired kidney function, and that measuring blood S-(2-succinyl)cysteine levels may be a minimally invasive way to evaluate the metabolic change in chronic kidney disease.


Nondestructive Measurement of the Evolution of Layer-Specific Mechanical Properties in Sub-10 nm Bilayer Films.
We use short wavelength extreme ultraviolet light to independently measure the mechanical properties of disparate layers within a bilayer film for the first time, with single-monolayer sensitivity. We show that in Ni/Ta nanostructured systems, while their density ratio is not meaningfully changed from that expected in bulk, their elastic properties are significantly modified, where nickel softens while tantalum stiffens, relative to their bulk counterparts. In particular, the presence or absence of the Ta capping layer influences the mechanical properties of the Ni film. This nondestructive nanomechanical measurement technique represents the first approach to date able to distinguish the properties of composite materials well below 100 nm in thickness. This capability is critical for understanding and optimizing the strength, flexibility and reliability of materials in a host of nanostructured electronic, photovoltaic, and thermoelectric devices.


Hypertension and renal complications in type 2 diabetes.
Diabetes mellitus and arterial hypertension are the leading causes of end-stage renal disease in industrialized countries. Although attention has focused on renal disease and insulin-dependent diabetes mellitus (type 1 diabetes), a silent epidemic of renal disease caused by type 2, noninsulin-dependent diabetes mellitus is rapidly developing. The course of renal function is heterogeneous in type 2 diabetic patients and reflects heterogeneous patterns of renal lesions. A subset of patients with microalbuminuria and proteinuria is characterized by the typical diabetic glomerulopathy usually observed in type 1 diabetic patients, with altered albumin excretion rate (AER), for example, glomerular basement membrane thickening and mesangial fractional volume expansion. These patients also have diabetic retinopathy and rapidly lose renal function despite tight blood pressure control. In contrast, a second subset of type 2 diabetic patients has normal or near-normal patterns of glomerular structure, despite abnormalities of AER comparable to those of the patients with diabetic glomerulopathy. Thus abnormalities of AER have a different renal prognostic value depending on the underlying renal structure. The patients with worse clinical prognosis and typical diabetic glomerulopathy also have diabetic retinopathy, whereas those with a better course of renal function quite often have no diabetic retinopathy. Several findings, albeit not unanimous, suggest that HbA1c levels above 7.5 to 8.0 % are closely associated with a rapid decay of renal function in type 2 diabetes. Tight blood pressure control plays a further important role in determining the progression of renal a damage in type 2 diabetes mellitus. Convincing evidence has been provided that drugs capable of inhibiting the renin-angiotensin hormonal system are quite effective in preventing and delaying the evolution of renal damage in both type 1 and 2 diabetes. Equally strong data support the view that other antihypertensive compounds such as beta-blockers and calcium antagonists also delay the progression of renal damage in diabetes mellitus. Whatever the drug used to treat hypertension, the majority of the authors conclude that blood pressure levels should be maintained below 130/85 mmHg in diabetic patients. While it is well established that uncontrolled diabetes underlies the development of diabetic nephropathy, newer evidence suggests that genetically determined susceptibility to hyperglycemia-caused glomerular injury is also necessary. More information on this issue will help to design new therapeutical approaches to treat hypertension and renal complications in type 2 diabetes.


Impact of Modern Management Strategies on the Clinical Outcome of Patients With Low Rectal Cancer - A Retrospective, Monocentric Cohort Study.
The aim of this study was to retrospectively investigate the impact of intersphincteric resection (ISR) and Enhanced Recovery After Surgery (ERAS) protocols for rectal cancer. Since we implemented rectal ERAS protocol and ISR in 2016, we retrospectively assessed and compared clinical, pathological and survival outcomes of two groups of patients: group 1, treated 2000-2015 (n=242); and group 2, treated 2016-2020 (n=108). Propensity score matching using nearest-neighbor method was used to match each patient of group 1 to a patient of group 2. Before and after matching, the American Society of Anesthesiology score for patients in group 1 was significantly lower than in group 2 (score of 3: 9.9% vs. 25.9%, p<0.0001) as were grade I-II complications (27.7% vs. 45.4% p<0.001). Before and after matching, the quality of the mesorectum excision was significantly lower in group 1 (complete in 31% vs. 59.2% p<0.0001). After matching, 3-year overall survival for groups 1 and 2 were similar (88.2% vs. 92.6%; p=0.988). ERAS and ISR had no negative impact on the oncological outcome of our patients and increased the preservation of bowel continuity.


Volumetric analysis of intravoxel incoherent motion imaging for assessment of solitary pulmonary lesions.
Background Differentiating between malignant and benign solitary pulmonary lesions (SPLs) is challenging. Purpose To determine diagnostic performance of intravoxel incoherent motion-based diffusion-weighted imaging (DW-IVIM) in distinguishing malignant from benign SPLs, using histogram analysis derived whole-tumor and single-section region of interest (ROI). Material and Methods This retrospective study received institutional review board approval. A total of 129 patients with diagnosed SPLs underwent DW-IVIM and apparent diffusion coefficient (ADC). ADC, slow diffusion coefficient (D), fast diffusion coefficient (D*), and perfusion fraction (f) were calculated separately by outlining whole-tumor and single-section ROI. Inter-observer reliability was assessed by inter-class correlation coefficient (ICC). ADC and DW-IVIM parameters were analyzed using independent-sample T-test. Receiver operating characteristic (ROC) analysis was constructed to determine diagnostic performance. Multiple logistic regression was performed to identify independent factors associated with malignant SPLs. Results There were 48 benign SPLs found in 35 patients and 94 patients with lung cancer (LC). ICC for whole-tumor ROI (range, 0.89-0.95) was higher than that for single-section ROI (range, 0.61-0.71). Mean ADC and D were significantly lower in the malignant group. ADC and D 10th showed significantly higher AUC values than did mean ADC and D. D showed significantly higher diagnostic accuracy in mean, 10th, and 25th percentiles than ADC values (all Ps < 0.05). D 10th was found to be an independent factor in discriminating LCs with an odds ratio of -1.217. Conclusion Volumetric analysis had higher reproducibility and diagnostic accuracy than did single-section. Further, compared to ADC, D value differentiated benign SPLs from LCs more accurately.


Insulin activates furosemide-sensitive K+ and Cl- uptake system in BC3H1 cells.
Insulin augments the activity of Na(+)-K(+)-adenosinetriphosphatase (ATPase) in skeletal muscles. This study shows that when furosemide- and bumetanide-inhibitable 86Rb+ uptake is measured in the skeletal muscle-like BC3H1 cell line, insulin and insulin-like growth factor I (IGF-I) activate a loop diuretic-sensitive K+ and Cl- transport system but have no effect on Na(+)-K(+)-ATPase. The insulin-stimulated K+ transport system is extracellular Na+ concentration ([Na+]o) independent and extracellular Cl- concentration ([Cl-]o) dependent. Na(+)-independent K(+)-Cl- cotransport systems have been identified in other cells, but their sensitivity to insulin or growth factors has not been described. The affinities of the insulin-stimulated K+ uptake in BC3H1 cells for K+ (0.9 +/- 0.1 mM) and loop diuretics (5.9 x 10(-7) and 10(-7) M for furosemide and bumetanide, respectively) are higher than those of K(+)-Cl- cotransporters in other cells. Thus the insulin-stimulated K+ and Cl- transport system in BC3H1 seems kinetically different from K(+)-Cl- cotransporters in other cells. Insulin and IGF-I may activate a unique K(+)-Cl- cotransporter or activate a [Na+]o-independent K(+)-Cl- cotransport mode of Na(+)-K(+)-Cl- cotransporter in BC3H1 cells.


Biochemical and cellular basis for circadian rhythms in obstructive lung disease and implications for theophylline therapy.
In the majority of patients one of the primary features of bronchial asthma is the occurrence of nocturnal symptoms. There are significant bioperiodicities for hormonal, neural, cellular and humoral factors and for mediators, which are all in favour of reducing bronchial patency during the night. In the morning hours between 2 and 6 a.m. the histamine concentration shows a peak, the adrenaline and cyclic AMP have their minimum, while the cortisol secretion with its delayed onset of action is already ascending. The kallikrein kinin system is activated by histamine. Circadian variations have also been found for the receptor density of beta-receptors, cAMP, adenylcyclase and phosphodiesterase. While circadian rhythms are not known for the platelet activating factor (PAF) at the present time, the known nocturnal peak of thromboxane A2 may influence PAF-liberation as well. The bronchodilating metabolite of the cyclooxygenase pathway, PGE2, was found to have depressed levels during the night. Total plasma, total protein as well as IgA, IgM, IgG and IgE have a minimum during the night, cellular elements like T11-, T4-, B-lymphocytes and Leu8 have a maximum. Slow release theophylline is today the most important drug for nocturnal asthma. Theophylline is known to interfere with histamine release from mast cells, mediator release of arachidonic acid metabolites, the cyclic AMP concentration and suppressor cell activity. Important work remains to be done to clarify the therapeutic and immunological role of theophylline in nocturnal asthma and to identify the subgroups of patients having the greatest benefit of theophylline treatment.


Occurrence of atrial fibrillation in patients with paroxysmal supraventricular tachycardia due to atrioventricular nodal reentry.
The occurrence of atrial fibrillation in patients with paroxysmal supraventricular tachycardia (PSVT) has been well documented when PSVT is secondary to atrioventricular reentry, but not when PSVT is secondary to atrioventricular nodal reentry (AVNRT). Seventeen patients with AVNRT were followed using transtelephonic electrocardiogram monitoring to document symptomatic tachycardias. The median length of telephone monitor surveillance was 357 days. Fifteen of 17 patients transmitted electrocardiograms that showed PSVT. Three of 17 patients (18%) transmitted electrocardiograms that showed atrial fibrillation. A transition from PSVT into atrial fibrillation was not recorded, but all three did have PSVT recorded on other days of follow-up. We report the occurrence of atrial fibrillation in patients with AVNRT and that its incidence is higher than expected for the general population.


Cloudy with a Chance of Insights: Context Dependent Gene Regulation and Implications for Evolutionary Studies.
Research in various fields of evolutionary biology has shown that divergence in gene expression is a key driver for phenotypic evolution. An exceptional contribution of cis-regulatory divergence has been found to contribute to morphological diversification. In the light of these findings, the analysis of genome-wide expression data has become one of the central tools to link genotype and phenotype information on a more mechanistic level. However, in many studies, especially if general conclusions are drawn from such data, a key feature of gene regulation is often neglected. With our article, we want to raise awareness that gene regulation and thus gene expression is highly context dependent. Genes show tissue- and stage-specific expression. We argue that the regulatory context must be considered in comparative expression studies.


Attachment style, self-concealment, and interpersonal distance among Japanese undergraduates.
The relationships among attachment style, self-concealment, and interpersonal distance were studied with 71 Japanese undergraduates (33 men and 38 women, ages 18 to 20 years, M = 18.7, SD= .6). Participants completed a questionnaire about Self-concealment and Attachment Styles (Secure, Anxious, and Avoidant). One week later, Interpersonal Distance, which individuals maintain between themselves and others, was measured by the stop-distance paradigm. Analysis showed that scores for more Anxious and Avoidant Attachment Styles were positively correlated with those for greater Self-concealment. Scores for greater Self-concealment and more Anxious Attachment Style were also correlated with longer Interpersonal Distance.


Cerebrospinal fluid-contacting neurons affect the expression of endogenous neural progenitor cells and the recovery of neural function after spinal cord injury.
To explore the relationship between cerebrospinal fluid-contacting neurons (CSF-cNs) and endogenous neural progenitor cells (ENPCs) and whether CSF-cNs are involved in nerve repair after spinal cord injury (SCI). Cholera toxin B-horseradish peroxidase complex (CB-HRP) and cholera toxin B conjugated with saporin (CB-SAP) were injected into the lateral ventricles of spinal cord injured rats to mark and destroy the CSF-cNs. Then the rats in the experimental group were injured by SCI. Observe the content and co-expression of CSF-cNs and ENPCs in rats of each group, and observe the recovery of motor function after SCI in each group. After the destruction of CSF-cNs, the number of ENPCs decreased significantly in the long term after the surgery, and the recovery of motor function also deteriorated as compared to the group with intact CSF-cNs. Meanwhile some cells in the spinal cord express both the biological marker of CSF-cNs and ENPCs. This study shows that the population of ENPCs and motor function recovery in SCI rats declined after the destruction of CSF-cNs, suggesting that CSF-cNs affect the ENPCs population and may be involved in the recovery of neural function after SCI.


Clinical significance and prognostic value of serum autoantibody tests in multiple sclerosis.
It is known that multiple sclerosis (MS) often coexists with other autoimmune diseases. Hence, autoantibody (auto-Ab) tests may prove useful in the differential diagnosis of MS. The objectives of this study were to: (a) investigate the prevalence of auto-Ab positivity at the beginning of the MS diagnostic process; (b) assess whether Auto-Ab+ and Auto-Ab- patients differ in baseline clinical, laboratory, and radiological parameters; and (c) investigate the prognostic value during a two-year follow-up period. This retrospective study consisted of 450 patients aged between 18 and 55 years. All patients underwent a wide range of auto-Ab tests, anti-nuclear antibody (ANA) tests in particular. The expanded disability status scale (EDSS) scores of the patients were recorded at the time of diagnosis and at the end of a two-year follow-up period. The mean age of the 212 patients, 148 (69.8%) female and 64 (30.2%) male, included in the study sample was 37 ± 10.83 years. The rate of relapsing cases was 84% (178). Oligoclonal band (OCB) was positive in 142 (86.6%) of the 164 tested cases. At least one of the auto-Ab tests was positive in 51 (24.1%) of the cases. ANA test was positive in 21 (9.9%) cases. There was no significant difference between patients with at least one positive auto-Ab test and without any positive auto-Ab test and between ANA-positive and ANA-negative patients in terms of age, gender, clinical features of MS, presence of brain stem lesion, presence of spinal lesion, OCB positivity, level of clinical improvement after the first pulse steroid treatment, family history, presence of comorbidity, presence of autoimmune disease, or EDSS scores recorded at the end of the two-year follow-up period (p > 0.05). Our study findings revealed that Auto-Ab positivity was more common in MS patients than in the general population. However, given their limited contribution to the diagnosis and differential diagnosis of MS with no effect on the prognostic process, auto-Ab tests should be requested only in the event of accompanying autoimmune disease symptoms, and in cases where the diagnosis of MS may be suspected.


Vitamin D deficiency in girls from South Brazil: a cross-sectional study on prevalence and association with vitamin D receptor gene variants.
Vitamin D deficiency has been associated with a multitude of disorders including diabetes, defective insulin secretion as well as rickets and poor bone health. Vitamin D is also a concern during childhood and adolescence and has been reported in girls from South Brazil. We determined the prevalence of vitamin D deficiency in girls from South Brazil and investigated whether the genotypic distribution of the BsmI, ApaI and TaqI polymorphisms of the VDR gene and their haplotypes were associated with vitamin D levels. Cross-sectional study including 234 apparently healthy girls aged 7 to 18 years. Height and weight were measured for calculation of body mass index (BMI) percentiles for age. Plasma levels of 25-hydroxyvitamin D [25(OH)D] were assessed. Participants were genotyped for ApaI (rs7975232), TaqI (rs731236), and BsmI (rs1544410) SNPs. The median and interquartile range (25-75%) of BMI percentile was 62.0 (33.3 - 84.9). The frequency of overweight/obesity was 24.9%. Circulating levels of 25(OH)D (≥ 30 ng/mL) were adequate in 9.4%; insufficient in 54.3% (20-29 ng/mL); and deficient in 36.3% (< 20 ng/mL). Genotype frequencies were GG = 47.0%, GA = 41.5%, and AA = 11.5% for BsmI; GG = 16.7%, GT = 52.6%, and TT = 30.8% for ApaI; TT = 46.2%, TC = 44.9% and CC = 9.0% for TaqI. Genotypes with no gene variance (ancestral wild genotype) of BsmI (GG vs. GA + AA, two-tailed Student's t-test p < 0.001), ApaI (GG vs. GT + TT, two-tailed Student's t-test p = 0.031) and TaqI (TT vs. TC + CC, two-tailed Student's t-test p = 0.005) SNPs and the GGT haplotype (two-tailed Student's t-test p = 0.036) were significantly associated with lower 25(OH)D levels. 25-hydroxyvitamin D deficiency and insufficiency were highly prevalent in this sample. The BsmI, ApaI and TaqI wild variants of the VDR gene, as well as the GGT haplotype, were associated with lower vitamin D levels, suggesting that VDR gene polymorphisms could be linked to higher susceptibility to vitamin D deficiency in a sub-population of children and adolescents.