Serving the enterprise and beyond with informatics for integrating biology and the bedside (i2b2).
Informatics for Integrating Biology and the Bedside (i2b2) is one of seven projects sponsored by the NIH Roadmap National Centers for Biomedical Computing (http://www.ncbcs.org). Its mission is to provide clinical investigators with the tools necessary to integrate medical record and clinical research data in the genomics age, a software suite to construct and integrate the modern clinical research chart. i2b2 software may be used by an enterprise's research community to find sets of interesting patients from electronic patient medical record data, while preserving patient privacy through a query tool interface. Project-specific mini-databases ("data marts") can be created from these sets to make highly detailed data available on these specific patients to the investigators on the i2b2 platform, as reviewed and restricted by the Institutional Review Board. The current version of this software has been released into the public domain and is available at the URL: http://www.i2b2.org/software.


N-Acetylcysteine reverses silver nanoparticle intoxication in rats.
The increasing use of silver nanoparticles (AgNPs) in consumer products raises the risk of human toxicity. Currently, there are no therapeutic options or established treatment protocols in cases of AgNPs intoxication. We demonstrated previously that thiol antioxidants compounds can reverse the cytotoxicity induced by AgNPs in Huh-7 hepatocarcinoma cells. Here, we investigated the use of N-acetylcysteine (NAC) against the systemic toxic effects of AgNPs (79.3 nm) in rats. Biochemical, histopathological, hematological, and oxidative parameters showed that a single intravenous injection of AgNPs (5 mg/kg b.w.) induced deleterious effects such as hepatotoxicity, potentially as a result of AgNPs accumulation in the liver. Treatment with a single intraperitoneal injection of NAC (1 g/kg b.w.) one hour after AgNPs exposure significantly attenuated all toxic effects evaluated and altered the bioaccumulation and release patterns of AgNPs in rats. The findings show that NAC may be a promising candidate for clinical management of AgNPs intoxication.


Respiratory health survey of respiratory therapists.
The purpose of this study was to determine whether respiratory therapists (RTs) had an elevated risk of respiratory symptoms and to determine the association of work exposures with symptoms. Mailed questionnaire responses from 275 RTs working in British Columbia, Canada, were compared to those of 628 physiotherapists who had been surveyed previously. Analyses incorporated logistic regression analysis with adjustment for age, sex, smoking status, and childhood asthma. Compared to physiotherapists, RTs had over twice the risk of being woken by dyspnea, having wheeze, asthma attacks, and asthma diagnosed after entering the profession. Among RTs, two work factors associated with asthma were sterilizing instruments with glutaraldehyde-based solutions and the use of aerosolized ribavirin. RTs who used an oxygen tent or hood had the highest risk of asthma diagnosed after entering the profession (odds ratio [OR], 8.3; 95% confidence interval [CI], 12.6 to 26.0) and of asthma attacks in the last 12 months (OR, 3.6; 95% CI, 1.2 to 10.9). Our data suggest that RTs may be at an increased risk for asthma-like symptoms and for receiving a diagnosis of asthma since starting to work in their profession, possibly related to exposure to glutaraldehyde and aerosolized ribavirin.


Multiple forms of facilitation produced by aversive tentacular stimuli in cerebral ganglion sensory neurons of Aplysia.
Aversive tentacular stimuli can produce both nonassociative and associative modification of head-waving behavior of Aplysia. Sensory neurons (the J/K cluster SNs) in the cerebral ganglion of Aplysia constitute an afferent pathway for aversive stimuli of the anterior tentacles. We used intracellular recording to examine plasticity in these neurons, particularly side-specific or site-specific alterations, which may be involved in mediating aspects of the learning induced by tentacle shock. The results of these experiments indicate that the J/K SNs exhibit several forms of plasticity: (1) Post-tetanic potentiation (PTP); (2) heterosynaptic facilitation; (3) activity-dependent synaptic facilitation; and (4) side-specific spike broadening. Furthermore, by activating SNs directly to produce PTP and producing heterosynaptic facilitation with tentacular stimuli that were either inside or outside the receptive field of individual SNs, it was possible to dissect and analyze the differential contribution of intrinsic SN activity and heterosynaptic modulation to activity-dependent facilitation induced by behaviorally relevant stimuli. Collectively, these data raise the possibility that plasticity in primary afferent SNs may be involved in US processing during learning induced by tentacle stimulation.


A Budding Relationship: Bacterial Extracellular Vesicles in the Microbiota-Gut-Brain Axis.
The discovery of the microbiota-gut-brain axis has revolutionized our understanding of systemic influences on brain function and may lead to novel therapeutic approaches to neurodevelopmental and mood disorders. A parallel revolution has occurred in the field of intercellular communication, with the realization that endosomes, and other extracellular vesicles, rival the endocrine system as regulators of distant tissues. These two paradigms shifting developments come together in recent observations that bacterial membrane vesicles contribute to inter-kingdom signaling and may be an integral component of gut microbe communication with the brain. In this short review we address the current understanding of the biogenesis of bacterial membrane vesicles and the roles they play in the survival of microbes and in intra and inter-kingdom communication. We identify recent observations indicating that bacterial membrane vesicles, particularly those derived from probiotic organisms, regulate brain function. We discuss mechanisms by which bacterial membrane vesicles may influence the brain including interaction with the peripheral nervous system, and modulation of immune activity. We also review evidence suggesting that, unlike the parent organism, gut bacteria derived membrane vesicles are able to deliver cargo, including neurotransmitters, directly to the central nervous system and may thus constitute key components of the microbiota-gut-brain axis.


Prevalence of enduring and disabling mental illness in the inner city.
Previous research identifying the long-term mentally ill in primary care has been outside areas of deprivation. We used a case finding approach by a primary care group to identify the prevalence and characteristics of people with enduring and disabling mental ill health in a disadvantaged inner-city community. We found a high point prevalence (12.9 per 1000 patients) of enduring psychotic and non-psychotic illness (36.1% and 63.9% respectively). This contributed to considerable workload and disability, and included a significant proportion of older people (24.6% aged over 65 years). The approach may be useful for local needs assessment. It highlights a need to consider disability as well as diagnosis for service development.


Prevalence, patterns and correlates of smokeless tobacco use in Nigerian adults: An analysis of the Global Adult Tobacco Survey.
The global tobacco epidemic contributes to more than 8 million deaths annually. However, most tobacco control interventions have been driven by an emphasis on smoked tobacco. Globally and more so in Nigeria, less attention has been paid to the similarly harmful smokeless tobacco (SLT) whose use appeals to a different demography. We examined the prevalence, patterns of use and correlates of SLT in Nigerian adults to guide targeted control efforts. We conducted a secondary analysis of the 2012 Global Adult Tobacco Survey (GATS) data. We obtained data on 9,765 non-institutionalised adults aged 15 years and older. Variables included current SLT use, sociodemographic characteristics and perceived harm of SLT use. We used Chi-square test to examine associations and binary logistic regression to assess predictors of current SLT use. All analyses were conducted with sample-weighted data. The prevalence of current SLT use was 1.9% of all adults. About 1.4% were daily users. The main types were snuff by nose (1.6%) and snuff by mouth (0.8%). There were higher odds of current SLT use for those in the South-East region (aOR = 13.99; 95% CI: 4.45-43.95), rural area residents (aOR = 1.56; 95% CI: 1.04-2.35), males (aOR = 4.43; 95% CI: 2.75-7.11), the 45-64 years age-group (aOR = 10.00; 95% CI: 4.12-24.29), those with no formal education (aOR = 2.67; 95% CI: 1.01-7.05), and those with no perception of harm from SLT use (aOR = 3.81, 95% CI: 2.61-5.56). The prevalence of SLT use among Nigerian adults was low with clearly identified predictors. While a majority were aware of harm from SLT use, an unacceptably high proportion remain unaware. We recommended targeted interventions to increase awareness of the harmful effects of SLT use especially among residents of the South-East, those in rural areas, males, and individuals with no formal education. We also recommended a follow-up survey.


Prevention of anaemia in pregnancy using insecticide-treated bednets and sulfadoxine-pyrimethamine in a highly malarious area of Kenya: a randomized controlled trial.
To compare the effects of intermittent treatment with sulfadoxine-pyrimethamine (SP) given during the second and third trimester of pregnancy, the use of insecticide-treated nets (ITN), or the combination of both on haemoglobin (Hb) levels during pregnancy, a randomized, placebo-controlled intervention trial was conducted in a malaria-endemic area of western Kenya from July 1997 to September 1999. Primigravidae and secundigravidae were enrolled into the study and randomized into 4 intervention groups: (i) ITNs and SP, (ii) ITNs and placebo SP, (iii) SP alone, and (iv) placebo SP. All groups were offered case management and iron and folic acid supplementation. Seven hundred and fifty-two women were followed until delivery (53.2% were primigravidae and 46.8% secundigravidae). Among primigravidae in all the groups there was a significant improvement in Hb levels at delivery (107.6 g/L) compared with recruitment (101.9 g/L) (P < 0.006) with the greatest improvement in the combination ITNs + SP group. The protective efficacy of ITNs + SP on anaemia was 55.8% (95% CI 30.6-71.8), of SP alone 50.9% (95% CI 22.2-69.0), and of ITNs 41.6% (95% CI 9.8-62.3). Among secundigravidae, Hb levels were slightly lower at delivery compared with recruitment (P = 0.03). It was concluded that malaria is a major cause of anaemia in primigravidae but that other causes play a more significant role in secundigravidae, and that intermittent treatment with SP or use of ITNs benefits primigravidae more than secundigravidae.


Farmers' perceptions, knowledge, and management of coffee pests and diseases and their natural enemies in Chiapas, Mexico.
Small farmers' perceptions of coffee Coffea arabica L. herbivores and their natural enemies, how those perceptions relate to field infestation levels, and pest management practices being implemented by members from two organic and nonorganic coffee grower organizations in the Soconusco region, southeastern Mexico, were analyzed through an interview survey, diagnostic workshops, and field sampling. The terms pest, disease, and damage were commonly used as synonyms. The major phytophagous species, as perceived by the interviewees, were Hypothenemus hampei (Ferrari), and to a lesser extent the fungi Corticium koleroga Cooke (Höhnel) and Hemileia vastatrix Berkeley & Broome. Among the nonorganic farmers, other nonpest-related constraints were regarded as more important. Awareness of the existence of natural enemies was low, despite more organic farmers have used the ectoparasitoid bethylid Cephalonomia stephanoderis Betrem against H. hampei. Labor supplied by household members was most frequent for pest control; only organic farmers exchanged labor for this purpose. The levels of infestation by H. hampei, Leucoptera coffeella Guérin-Méneville, and C. koleroga were lower within the organic coffee stands. However, a low effectiveness for pest control was commonly perceived, probably due to a feeling, among the organic farmers, of a low impact of their pest management extension service, whereas a lack of motivation was prevalent among the nonorganic farmers, shown by a concern with their low coffee yields and the emigration of youth. The importance of understanding farmers' perceptions and knowledge of pests and their natural enemies and the need for participatory pest management approaches, are discussed.


Expectations and strategies regarding islet transplantation: metabolic data from the GRAGIL 2 trial.
Whether islet transplantation should be aimed at restoring insulin independence or providing adequate metabolic control is still debated. The GRAGIL2 trial was designed as a phase 1-2 study where primary outcome was the rate of insulin independence, and secondary outcome was the success rate defined by a composite score based upon basal C-peptide, HbA1c, hypoglycemic events, and exogenous insulin needs. C-peptide negative type 1 brittle diabetic patients experiencing severe hypoglycemia were eligible to receive a maximum of two islet preparations totalizing 10,000 IE/kg or more, with a threshold of 5,000 IE/kg for the first infusion, according to the Edmonton protocol, within the Swiss-French GRAGIL multicentric network. A sequential analysis with a triangular test was performed in every five patients after 6- and 12-month follow-up. Maximal inefficiency was set at 40% and minimal efficiency at 66%. From September 2003 to October 2005, 10 patients were included. Median waiting time was 6.7 months (first injection) and 9 weeks (second injection). All but one patient received 11,089+/-505 IE/kg: one received a single graft of 5398 IE/kg. At 6 months, insulin independence and composite success rates were 6 of 10 and 6 of 10, respectively. At 12 months, insulin independence was observed in 3 of 10 patients and success in 5 of 10 patients. Based upon our sequential analysis settings, islet transplantation failed to achieve the primary goal, insulin independence, but tended to succeed in reaching the secondary goal, successful metabolic control. Currently it appears to be a successful biological closed-loop glucose control method for brittle diabetes.


Association between urinary organophosphate flame retardant diesters and steroid hormones: A metabolomic study on type 2 diabetes mellitus cases and controls.
Metabolomic analysis was conducted by collecting urine samples from 128 participants in diagnose of type 2 diabetes mellitus (T2DM) and 105 volunteers in healthy condition, in order to identify biomarkers of experimental populations. The urinary concentrations of organophosphate flame retardant (OPFR) diesters were determined and linear regression model was used to find associations between OPFR diesters and the identified biomarkers. The urinary concentrations of OPFR diesters ranged from 0.17-779 μg/g creatinine. Diphenyl phosphate (DPHP) was detected with the highest frequency of 97% at a median level of 1.21 μg/g, and bis(1-chloro-2-propyl) phosphate (BCIPP) dominated the highest median level at 4.24 μg/g with a detection frequency of 94.4%. As compared with the control, the urinary median concentrations of bis(2-butoxyethyl) phosphate (BBOEP), bis(1,3-dichloro-2-propyl) phosphate (BDCPP) and DPHP were 2.76, 2.48, and 1.46 times higher in people with T2DM, respectively. Urinary metabolomic data revealed that steroid synthesis was the most significantly altered metabolic pathway between the case and control population. Two biomarkers of cortisol and cortisone that play an important role in steroid hormone regulation were quantified. The linear regression model indicated that per-quartile range increase in the concentrations of each OPFR diester was associated 18%-41% increase in the concentrations of cortisol and cortisone, which may impact energy metabolism linked with T2DM. To our knowledge, this study for the first time reported the altered levels of steroid hormones associated with urinary OPFR diesters.


Occupational Stress Management and Burnout Interventions in Nursing and Their Implications for Healthy Work Environments: A Literature Review.
This article reports on a literature review of workplace interventions (i.e., creating healthy work environments and improving nurses' quality of work life [QWL]) aimed at managing occupational stress and burnout for nurses. A literature search was conducted using the keywords nursing, nurses, stress, distress, stress management, burnout, and intervention. All the intervention studies included in this review reported on workplace intervention strategies, mainly individual stress management and burnout interventions. Recommendations are provided to improve nurses' QWL in health care organizations through workplace health promotion programs so that nurses can be recruited and retained in rural and northern regions of Ontario. These regions have unique human resources needs due to the shortage of nurses working in primary care.


Treatment of unruptured aneurysms with GDCs: clinical experience with 247 aneurysms.
The GDC system is a valuable therapeutic alternative to surgical treatment of intracranial aneurysms. We present our clinical experience with the GDC technique used to treat unruptured cerebral aneurysms. This is a retrospective review of 217 patients with 247 unruptured intracranial aneurysms who underwent embolization with GDCs between August 1991 and June 2000. One hundred sixty-seven of the 217 were female patients. Patient age ranged from 13 to 83 years. In 118 patients, the aneurysms were found when unrelated non-neurologic conditions indicated angiography, CT angiography, or MR angiography (group 1). Fifty-one patients with mass effect symptoms comprised group 2, 12 patients with aneurysms associated with arteriovenous malformations or tumors of the brain comprised group 3, 17 patients with unruptured aneurysms treated during the chronic phase of subarachnoid hemorrhage (SAH) comprised group 4, and 19 patients treated during the acute phase of SAH due to another ruptured aneurysm comprised group 5. Angiographic results revealed complete occlusion in 138 of 247 aneurysms (55.9%), neck remnants in 92 (37.2%), and incomplete occlusion in three (1.2%). GDC embolization was attempted unsuccessfully in 14 aneurysms (5.7%). Of the 198 patients without acute SAH (groups 1-4), 186 (93.9%) of 198 remained neurologically unchanged. Eleven of the 217 patients (5.1%) experienced neurologic deterioration caused by immediate procedural complications. One patient died (0.5%) as a result of aneurysmal rupture during embolization. For asymptomatic patients and those treated after the chronic period of SAH, the mean hospitalization stay was 2.9 days. GDC technology constitutes safe treatment for unruptured aneurysms, with successful anatomic and clinical results and low complication rates achieved.


Functional, biochemical and genetic diversity of prokaryotic nitrate reductases.
Prokaryotic nitrate reduction can serve a number of physiological roles and can be catalysed by a number of biochemically distinct nitrate reductases. Three distinct nitrate reductase classes can be indentified in prokaryotes, NAS, NAR and NAP. NAS is located in the cytoplasmic compartment and participates in nitrogen assimilation. NAR is usually a three-subunit complex anchored to the cytoplasmic face of the membrane with its active site located in the cytoplasmic compartment and is involved in anaerobic nitrate respiration. NAP is a two-subunit complex, located in the periplasmic compartment, that is coupled to quinol oxidation via a membrane anchored tetraheme cytochrome. It shows considerable functional flexibility by participating in anaerobic respiration or redox energy dissipation depending on the organism in which it is found. The members of all three classes of enzymes bind the bis-molybdopterin guanine dinucleotide cofactor at the active site, but they differ markedly in the number and nature of cofactors used to transfer electrons to this site. Analysis of prokaryotic genome sequences available at the time of writing reveals that the different nitrate reductases are phylogenetically widespread.


The influence of electrical stimulation in vitro on protein synthesis and other metabolic parameters of rat extensor digitorum longus muscle.
1. Apparatus is described in which rat extensor digitorum longus muscle can be incubated in buffer under conditions of light tension and be subject to contractures induced by electrical stimulation in vitro. Under these conditions the tissue retains its weight, its content of potassium and size of the extracellular space at values similar to those in vivo. 2. Though uptake of glucose was enhanced on addition of insulin, there was little increase in glucose consumption on stimulation. Breakdown of glycogen and enhancement of lactate output were found on stimulation. 3. Incorporation into protein of several labelled amino acids was diminished during stimulation. Accumulation of [(14)C]leucine was enhanced whereas that of glycine was decreased. 4. There were no very consistent changes in the content of free unlabelled amino acids during incubation with or without stimulation. Comparison of actual amino acid concentrations in tissue and incubation mixture with accumulation of (14)C-labelled amino acid indicated that full equilibration of the cell pool of amino amino acids with the medium is slow. 5. Substantial oxidation of several (14)C-labelled acids was observed. 6. The ATP content of the tissue declined a little during incubation and somewhat faster after a period of stimulation. 7. The results are discussed in relation to the way in which exercise can induce muscle hypertrophy.


The selenium content of SEPP1 versus selenium requirements in vertebrates.
Selenoprotein P (SEPP1) distributes selenium (Se) throughout the body via the circulatory system. For vertebrates, the Se content of SEPP1 varies from 7 to 18 Se atoms depending on the species, but the reason for this variation remains unclear. Herein we provide evidence that vertebrate SEPP1 Sec content correlates positively with Se requirements. As the Se content of full length SEPP1 is genetically determined, this presents a unique case where a nutrient requirement can be predicted based on genomic sequence information.


Point-of-Care Testing in Neurosurgery.
Coagulation disorders can have a major impact on the outcome of neurosurgical patients. The central nervous system is located within the closed space of the skull, and therefore, intracranial hemorrhage can lead to intracranial hypertension. Acute brain injury has been associated with alterations of various hemostatic parameters. Point-of-care (POC) techniques such as rotational thromboelastometry are able to identify markers of coagulopathy which are not reflected by standard assessment of hemostasis (e.g., hyperfibrinolysis). In patients with acute brain injury, POC test results have been associated with important outcome parameters such as mortality and need for neurosurgical intervention. POC devices have also been used to rapidly identify and quantify the effects of antithrombotic medication. In cases of life-threatening intracranial hemorrhage, this information can be valuable when deciding over administration of prohemostatic substances or immediate neurosurgical intervention. In elective neurosurgical procedures, POC devices can provide important information when unexpected bleeding occurs or in cases of prolonged operative time with subsequent blood loss. Initial experiences with POC devices in neurosurgical care have shown promising results but further studies are needed to characterize their full potential and limitations.


Measurement of absolute lung volumes by imaging techniques.
In this paper, the techniques available for estimating total lung capacities from standard chest radiographs in children and infants as well as adults are reviewed. These techniques include manual measurements using ellipsoid and planimetry techniques as well as computerized systems. Techniques are also available for making radiographic lung volume measurements from portable chest radiographs. There are inadequate data in the literature to support recommending one specific technique over another. Though measurements of lung volumes by radiographic, plethysmographic, gas dilution or washout techniques result in remarkably similar mean results when groups of normal subjects are tested, in patients with disease, the results of these different basic measurement techniques can differ significantly. Computed tomographic and magnetic resonance techniques can also be used to measure absolute lung volumes and offer the theoretical advantages that the results in individual subjects are less affected by variances of thoracic shape than are measurements made using conventional chest radiographs.


Deficiency of Shank2 causes mania-like behavior that responds to mood stabilizers.
Genetic defects in the synaptic scaffolding protein gene, SHANK2, are linked to a variety of neuropsychiatric disorders, including autism spectrum disorders, schizophrenia, intellectual disability, and bipolar disorder, but the molecular mechanisms underlying the pleotropic effects of SHANK2 mutations are poorly understood. We generated and characterized a line of Shank2 mutant mice by deleting exon 24 (Δe24). Shank2Δe24-/- mice engage in significantly increased locomotor activity, display abnormal reward-seeking behavior, are anhedonic, have perturbations in circadian rhythms, and show deficits in social and cognitive behaviors. While these phenotypes recapitulate the pleotropic behaviors associated with human SHANK2-related disorders, major behavioral features in these mice are reminiscent of bipolar disorder. For instance, their hyperactivity was augmented with amphetamine but was normalized with the mood stabilizers lithium and valproate. Shank2 deficiency limited to the forebrain recapitulated the bipolar mania phenotype. The composition and functions of NMDA and AMPA receptors were altered at Shank2-deficient synapses, hinting toward the mechanism underlying these behavioral abnormalities. Human genetic findings support construct validity, and the behavioral features in Shank2 Δe24 mice support face and predictive validities of this model for bipolar mania. Further genetic studies to understand the contribution of SHANK2 deficiencies in bipolar disorder are warranted.


Weight-Related Concerns as Barriers to Exclusive Breastfeeding at 6 Months.
Human milk is considered the gold standard for infant nutrition, but more data are needed that examine the constellation of weight-related concerns as barriers to exclusive breastfeeding. The aim of this study was to examine how mothers' concerns regarding their own and their infants' weight, as well as disordered eating behaviors, were associated with breastfeeding self-efficacy and exclusive breastfeeding at 6 months. A prospective, quantitative, and self-report online survey design was used. Participants included 206 women (88.30% White, 59.20% with graduate degrees), with a mean age of 33.04 years ( SD = 4.31 years) and a mean prepregnancy body mass index (BMI) of 24.80 kg/m2 ( SD = 5.50 kg/m2), who had given birth within the past 6 months. Participants who reported not exclusively breastfeeding at 6 months had significantly higher prepregnancy BMI ( p < .001), higher body dissatisfaction ( p = .003), more disordered eating ( p = .036), higher child weight concerns ( p < .001), and lower breastfeeding self-efficacy ( p < .001). Mediation modeling revealed a direct negative relationship between prepregnancy BMI and exclusive breastfeeding at 6 months ( p < .001). Indirect negative relationships between prepregnancy BMI and exclusive breastfeeding at six months via (a) body dissatisfaction, (b) disordered eating, and (c) child weight concern, as well as breastfeeding self-efficacy (entered as concurrent mediators), were all significant. Mothers' weight, body image and eating concerns, concern regarding their children's weight, and breastfeeding self-efficacy may constitute critical barriers to exclusive breastfeeding at 6 months. Interventions to improve breastfeeding duration and confidence should target maternal body image and eating concerns.


Differentiation of strains of Actinobacillus actinomycetemcomitans by arbitrarily primed polymerase chain reaction.
The present study describes a method for amplifying DNA in Actinobacillus actinomycetemcomitans by using short, synthetic oligonucleotides of random sequence as primers in the polymerase chain reaction. Genomic DNA from each of 20 human isolates of A. actinomycetemcomitans was successfully amplified in a thermal cycler with a single synthetic primer (GGGTAACGCC) and reproducibly produced 14 different DNA amplification profiles (amplitypes). A. actinomycetemcomitans isolates from the same subject revealed the same amplitype. The arbitrarily primed polymerase chain reaction appears to be useful in characterizing human isolates of A. actinomycetemcomitans for studies of epidemiology and bacterial transmission.


Modulatory effect of neoadjuvant chemotherapy on the prognosis of patients with breast cancer.
To evaluate the changes in biological markers ER, PR, HER2 and Ki67 in residual tumor after surgery for locally advanced breast cancer (LABC), and also to evaluate the outcome of breast cancer patients treated with neoadjuvant chemotherapy (NAC). 144 breast cancer patients treated with NAC at the Oncology Institute of Vojvodina, Serbia from 2011 to 2015 were included in this study. Changes in biologic markers ER,PR, HER2/neu and Ki-67 were evaluated at diagnostic core biopsy and at the final surgery tissue specimens. Of 144 patients pathological complete response was achieved in 17 (12%) and these were excluded from the study. Evaluated were 127 patients with residual tumor after the final surgery. A change in hormone receptor status (ER,PR) occurred in 9.4% of the patients (ER in 5%, PR in 14.5%) and HER2 status in 4.7% of the patients. ER and PR status change from negative to positive was associated with better overall survival (OS), but without statistical significance (p=0.16). Patients with conversion of HER2 status from negative to positive lived longer (65 vs 42 months). Furthermore, it was determined that HER2 change from negative to positive was associated with better OS (p=0.03). Ki-67 changed in 17 (11.8%) patients. The decrease of Ki-67 expression after NAC was associated with better outcome. Median follow up was 37.5 months (range 16.2-76.8). Changes in hormone receptor status, HER2 status and Ki-67 occurred after NAC in patients with LABC. A change from negative to positive hormone receptor status and HER2 status offers new treatment options, like endocrine therapy, and/or trastuzumab therapy for breast cancer patients. The decrease of Ki-67 expression after NAC was associated with better outcome.


Incremental fuzzy mining of gene expression data for gene function prediction.
Due to the complexity of the underlying biological processes, gene expression data obtained from DNA microarray technologies are typically noisy and have very high dimensionality and these make the mining of such data for gene function prediction very difficult. To tackle these difficulties, we propose to use an incremental fuzzy mining technique called incremental fuzzy mining (IFM). By transforming quantitative expression values into linguistic terms, such as highly or lowly expressed, IFM can effectively capture heterogeneity in expression data for pattern discovery. It does so using a fuzzy measure to determine if interesting association patterns exist between the linguistic gene expression levels. Based on these patterns, IFM can make accurate gene function predictions and these predictions can be made in such a way that each gene can be allowed to belong to more than one functional class with different degrees of membership. Gene function prediction problem can be formulated both as classification and clustering problems, and IFM can be used either as a classification technique or together with existing clustering algorithms to improve the cluster groupings discovered for greater prediction accuracies. IFM is characterized also by its being an incremental data mining technique so that the discovered patterns can be continually refined based only on newly collected data without the need for retraining using the whole dataset. For performance evaluation, IFM has been tested with real expression datasets for both classification and clustering tasks. Experimental results show that it can effectively uncover hidden patterns for accurate gene function predictions.


Activation of XerCD-dif recombination by the FtsK DNA translocase.
The FtsK translocase pumps dsDNA directionally at ∼5 kb/s and facilitates chromosome unlinking by activating XerCD site-specific recombination at dif, located in the replication terminus of the Escherichia coli chromosome. We show directly that the γ regulatory subdomain of FtsK activates XerD catalytic activity to generate Holliday junction intermediates that can then be resolved by XerC. Furthermore, we demonstrate that γ can activate XerCD-dif recombination in the absence of the translocase domain, when it is fused to XerCD, or added in isolation. In these cases the recombination products are topologically complex and would impair chromosome unlinking. We propose that FtsK translocation and activation of unlinking are normally coupled, with the translocation being essential for ensuring that the products of recombination are topologically unlinked, an essential feature of the role of FtsK in chromosome segregation.


Marginalization and tooth loss in older Mexican adults.
To examine the association between the level of marginalization and tooth loss in adults aged ≥50 years in Mexico. A retrospective cross‑sectional study was conducted on 2098 adults selected from communities presenting both high and medium levels of marginalization, with data obtained from the annual reports of the Epidemiological Surveillance System of Oral Pathologies 2019-2022. Analysis considered sociodemographic characteristics such as sex, age, and years of education. The prevalence of tooth loss was determined using the Decayed, Missing and Filled teeth (DMFT) index. Negative binomial regression was used to estimate rate ratio (RR) and 95% confidence intervals (CIs), using the presence of lost teeth as an outcome. Most (62.4%) participants had lost at least one tooth, of whom 60.3% had a medium level of marginalization, 28.8% a high level, and 10.9% a very high level. More adults who lived in high or very high marginalization communities had lost teeth [RR=1.19 (95% CI 1.08 - 1.30)] than those living in communities with a medium level of marginalization. More males [RR=1.19 (95% CI 1.09 - 1.31)], adults ≥70 years [RR=1.77 (95% CI 1.57 - 1.99)], people who were illiterate [RR=1.60 (95% CI 1.35 - 1.88)] or who had poor oral hygiene [RR=1.26 (95% CI 1.15 - 1.38)] had also lost teeth. High and very high marginalization was associated with tooth loss in adults ≥50 years in Mexico. Effective oral health strategies are required to prevent tooth loss, as are interventions to improve access to and quality of dental services for marginalized communities.


Combination therapy with dendritic cells and lenalidomide is an effective approach to enhance antitumor immunity in a mouse colon cancer model.
In this study, we investigated efficacy of lenalidomide in combination with tumor antigen-loaded dendritic cells (DCs) in murine colon cancer model. MC-38 cell lines were injected subcutaneously to establish colon cancer-bearing mice. After tumor growth, lenalidomide (50 mg/kg/day) was injected intraperitoneally on 3 consecutive days in combination with tumor antigen-loaded DC vaccination on days 8, 12, 16, and 20. The tumor antigen-loaded DCs plus lenalidomide combination treatment exhibited a significant inhibition of tumor growth compared with the other groups. These effects were associated with a reduction in immune suppressor cells, such as myeloid-derived suppressor cells and regulatory T cells, with the induction of immune effector cells, such as natural killer cells, CD4+ T cells and CD8+ T cells in spleen, and with the activation of cytotoxic T lymphocytes and NK cells. This study suggests that a combination of tumor antigen-loaded DC vaccination and lenalidomide synergistically enhanced antitumor immune response in the murine colon cancer model, by inhibiting the generation of immune suppressive cells and recovery of effector cells, and demonstrated superior polarization of Th1/Th2 balance in favor of Th1 immune response. This combination approach with DCs and lenalidomide may provide a new therapeutic option to improve the treatment of colon cancer.


Pseudomonas aeruginosa PA80 is a cystic fibrosis isolate deficient in RhlRI quorum sensing.
Pseudomonas aeruginosa uses quorum sensing (QS) to modulate the expression of several virulence factors that enable it to establish severe infections. The QS system in P. aeruginosa is complex, intricate and is dominated by two main N-acyl-homoserine lactone circuits, LasRI and RhlRI. These two QS systems work in a hierarchical fashion with LasRI at the top, directly regulating RhlRI. Together these QS circuits regulate several virulence associated genes, metabolites, and enzymes in P. aeruginosa. Paradoxically, LasR mutants are frequently isolated from chronic P. aeruginosa infections, typically among cystic fibrosis (CF) patients. This suggests P. aeruginosa can undergo significant evolutionary pathoadaptation to persist in long term chronic infections. In contrast, mutations in the RhlRI system are less common. Here, we have isolated a clinical strain of P. aeruginosa from a CF patient that has deleted the transcriptional regulator RhlR entirely. Whole genome sequencing shows the rhlR locus is deleted in PA80 alongside a few non-synonymous mutations in virulence factors including protease lasA and rhamnolipid rhlA, rhlB, rhlC. Importantly we did not observe any mutations in the LasRI QS system. PA80 does not appear to have an accumulation of mutations typically associated with several hallmark pathoadaptive genes (i.e., mexT, mucA, algR, rpoN, exsS, ampR). Whole genome comparisons show that P. aeruginosa strain PA80 is closely related to the hypervirulent Liverpool epidemic strain (LES) LESB58. PA80 also contains several genomic islands (GI's) encoding virulence and/or resistance determinants homologous to LESB58. To further understand the effect of these mutations in PA80 QS regulatory and virulence associated genes, we compared transcriptional expression of genes and phenotypic effects with isogenic mutants in the genetic reference strain PAO1. In PAO1, we show that deletion of rhlR has a much more significant impact on the expression of a wide range of virulence associated factors rather than deletion of lasR. In PA80, no QS regulatory genes were expressed, which we attribute to the inactivation of the RhlRI QS system by deletion of rhlR and mutation of rhlI. This study demonstrates that inactivation of the LasRI system does not impact RhlRI regulated virulence factors. PA80 has bypassed the common pathoadaptive mutations observed in LasR by targeting the RhlRI system. This suggests that RhlRI is a significant target for the long-term persistence of P. aeruginosa in chronic CF patients. This raises important questions in targeting QS systems for therapeutic interventions.


The accuracy of self-monitoring and its relationship to self-focused attention in dysphoria and clinical depression.
The accuracy with which dysphoric (Study 1) and clinically depressed (Study 2) individuals make self-regulatory judgments about their own performance in the absence of external feedback and the extent to which this relates to trait self-focused attention (SFA) were examined. Relative to objective criteria, both dysphoric and depressed participants showed a positive judgment bias, overestimating the number of trials they had performed correctly. Relative to control participants, the dysphoric and depressed groups showed a reduction in the extent of this positive bias in that they judged error trials more accurately and correct trials less accurately. Although the dysphoric and depressed groups both reported elevated trait SFA, this did not correlate significantly with accuracy of self-judgment on the performance-monitoring task. Implications for self-regulation models of depression are discussed.


Exposure biomarkers and risk from gluing and heating of polyurethane: a cross sectional study of respiratory symptoms.
To define the relation between exposure to polyurethane (PUR) glue, biomarkers of exposure and effect, and work related symptoms that occur at least once a week. In a cross sectional study, 152 workers and 14 clerks in a factory with exposure to sprayed and heated PUR glue containing 4, 4'-diphenylmethane (MDI) or 1,6-hexamethylene (HDI) di-isocyanate were examined with gas chromatography-mass spectrometry (GC-MS) for metabolites of MDI in plasma (P-MDX) and urine (U-MDX), 2,4- and 2, 6-toluene di-isocyanate (TDI; P-TDX, U-TDX) and HDI in plasma and urine, specific serum IgG (S-IgG-MDI, S-IgG-HDI, and S-IgG-TDI, respectively) and IgE (S-IgE-MDI). Work related symptoms of the eyes and airways (nose or lower airways, or both), and lung function were also evaluated. P-MDX was detected in 65% of the workers, U-TDX in 47%, HDX in none. Three per cent were positive for S-IgE-MDI, 33% for S-IgG-MDI, 32% for S-IgG-TDI, and 12% for S-IgG-HDI. A few clerks had metabolites, and some had antibodies. Most metabolites and immunoglobulins were slightly correlated-for example, P-MDX v S-IgG-MDI: r(s)=0.21. Workers who heated glue had increased P-MDX (odds ratio (OR)=12 for a value above the median) and S-IgG-MDI (OR=3.7), sprayers P-2,4-TDX (OR=6.2) and P-2,6-TDX (OR=16). Twenty six per cent of the workers had work related symptoms of the airways, 21% from the nose, 11% from the lower airways. Spraying of glue increased the risk of work related symptoms and slightly decreased lung function. U-MDX was associated with work related symptoms from the airways (OR=3.7) and P-2,6-TDX with work related symptoms from the lower airways (OR=6.6). S-IgG-MDI was related to work related symptoms from the airways (OR=2.6). There were relations between exposures to sprayed and heated PUR glue based on MDI and HDI, concentrations of metabolites of MDI and TDI in plasma and urine, specific IgG serum antibodies against MDI, TDI, and HDI, and work related symptoms.


Geographic distribution and inheritance of three cytoplasmic incompatibility types in Drosophila simulans.
Wolbachia-like microorganisms have been implicated in unidirectional cytoplasmic incompatibility between strains of Drosophila simulans. Reduced egg eclosion occurs when females from uninfected strains (type W) are crossed with males from infected strains (type R). Here we characterize a third incompatibility type (type S) which is also correlated with the presence of Wolbachia-like microorganisms. Despite the fact that the symbionts cannot be morphologically distinguished, we observed complete bidirectional incompatibility between R and S strains. This indicates that the determinants of incompatibility are different in the two infected types. S/W incompatibility is unidirectional and similar to R/W incompatibility. A worldwide survey of D. simulans strains showed that type S incompatibility was found only in insular populations which harbor the mitochondrial type SiI. Both W and R types were found among mainland and island populations harboring the worldwide mitochondrial type SiII. Type S incompatibility could be involved in the reinforcement of the geographical isolation of SiI populations.


Phenotypic stability of trp operon recombinant plasmids in Escherichia coli.
The recombinant plasmids RSF2124-trp and pSC101-trp were examined for their phenotypic stability in Escherichia coli W3110 and its derivatives under various culture conditions. RSF2124-trp and pSC101-trp were stable in a trpAE1 strain. In an amber mutant of the tryptophan repressor gene, RSF2124-trp was fairly stable, whereas pSC101-trp was unstable. All Trp- segregants from the pSC101-trp carrier had lost the entire plasmid. In a mutant carrying the tnaA mutation, RSF2124-trp was unstable in rich media. Most Trp- segregants that appeared under these conditions were deleted in trp genes as well as in the cI gene on the recombinant plasmid. pSC101-trp in this tnaA mutant was also unstable. All Trp- segregants had lost the plasmid. Studies of enzyme activities revealed that the greater the activity of anthranilate synthase and tryptophan synthase in bacteria, the more segregants tended to appear in the stability test. RSF2124 and pSC101 without the trp gene were completely stable in the same bacteria. The apparent instability of bacteria carrying the recombinant plasmid could be explained by the lower growth rate compared with bacteria carrying only the vector plasmid, resulting in the enrichment of Trp- bacteria during culture.


A new transmissible agent causing acute hepatitis, chronic hepatitis and cirrhosis in dogs.
There is a hepatitis of dogs which occurs in acute, persistent and chronic forms. Histological studies of spontaneous cases suggested that several apparently diverse hepatic diseases might be stages of one process. This was also implied by follow up studies and case histories: acute non-lethal episodes were followed later by the development of chronic hepatitis, cirrhosis and liver failure. Serum was taken and homogenates of liver were made from three field cases representing different putative temporal stages of the complex. These were injected into experimental dogs and a hepatitis was induced in all. The cytopathological and histological changes were the same in all animals and were identical to field cases. Acute lethal disease and persistent infections were produced. Two second passages were carried out and an identical condition was induced, characterised by recurrent episodes of subclinical hepatitis and persistent infection. It is suggested that the disease might be named canine acidophil cell hepatitis in view of the pathognomonic cytopathology. Specific morphological criteria have been established for this hepatitis.


Examining the Relationship Between Speech Recognition and a Spectral-Temporal Test With a Mixed Group of Hearing Aid and Cochlear Implant Users.
Purpose Audiology clinics have a need for a nonlinguistic test for assessing speech scores for patients using hearing aids or cochlear implants. One such test, the Spectral-Temporally Modulated Ripple Test Lite for computeRless Measurement (SLRM), has been developed for use in clinics, but it, as well as the related Spectral-Temporally Modulated Ripple Test, has primarily been assessed with cochlear implant users. The main goal of this study was to examine the relationship between SLRM and the Arizona Biomedical Institute Sentence Test (AzBio) for a mixed group of hearing aid and cochlear implant users. Method Adult hearing aid users and cochlear implant users were tested with SLRM, AzBio in quiet, and AzBio in multitalker babble with a +8 dB signal-to-noise ratio. Results SLRM scores correlated with both AzBio recognition scores in quiet and in noise. Conclusions The results indicated that there is a significant relationship between SLRM and AzBio scores when testing a mixed group of cochlear implant and hearing aid users. This suggests that SLRM may be a useful nonlinguistic test for use with individuals with a variety of hearing devices.


Australian Football League Injury Characteristics Differ Between Matches and Training: A Longitudinal Analysis of Changes in the Setting, Site, and Time Span From 1997 to 2016.
Injury surveillance has been used to quantify the scope of the injury burden in Australian football. However, deeper statistical analyses are required to identify major factors that contribute to the injury risk and to understand how these injury patterns change over time. To compare Australian Football League (AFL) injury incidence, severity, prevalence, and recurrence by setting, site, and time span from 1997 to 2016. Descriptive epidemiology study. A total of 15,911 injuries and medical illnesses recorded by team medical staff at each club were obtained from the AFL's injury surveillance system and analyzed using linear mixed models with 3 fixed effects (setting, time span, site) and 1 random effect (club). All types of injuries and medical illnesses were included for analysis, provided that they caused the player to miss at least 1 match during the regular season or finals. Five-season time spans (1997-2001, 2002-2006, 2007-2011, and 2012-2016) were used for comparisons. Incidence rates were expressed at the player level. Recurrences were recoded to quantify recurrent injuries across multiple seasons. Compared with training injuries, match injuries had a 2.8 times higher incidence per season per club per player (matches: 0.070 ± 0.093; training: 0.025 ± 0.043; P < .001). Match injuries resulted in 1.9 times more missed matches per club per season (matches: 17.2 ± 17.0; training: 9.1 ± 10.5; P < .001). and were more likely to be recurrences (matches: 11.6% ± 20.0%; training: 8.6% ± 21.8%; P < .001). From the 1997-2001 to 2007-2011 time spans, overall injury severity increased from a mean of 3.2 to 3.7 missed matches (P ≤ .01). For the most recent 2012-2016 time span, injuries resulted in 3.6 missed matches, on average. Hip/groin/thigh injuries had the highest incidence (0.125 ± 0.120) and prevalence (19.2 ± 16.4) rates, and recurrences (29.3% ± 27.9%) were 15% more likely at this site than any other injury site. The risks of match injuries are significantly higher than those of training injuries in the AFL. Compared with the 1997-2001 time span, injuries became more severe during the 2007-2011 time span.


POWASSAN VIRUS: MORPHOLOGY AND CYTOPATHOLOGY.
Powassan virus, a North American tickborne group B arbovirus, multiplied after simultaneous inoculation into bottles or tubes of virus and trypsinized suspension of continuous-line cultures of rhesus monkey kidney cells, strain LLC-MK2. Cytopathic effects comprising cell rounding and cytoplasmic vacuolation were first observed five days after inoculation. Mixture of Powassan antiserum with virus before inoculation into tissue cultures inhibited the appearance of cytopathic effects. Hemagglutinins for rooster erythrocytes, optimally at pH 6.4 and 22 degrees C., first appeared in tissue culture supernatant fluids four days after inoculation.Electron microscopic observation of thin sections of infected tissue culture cells showed virus particles 360-380 A.U. along outer cell membranes and edges of cytoplasmic vacuoles. In phosphotungstic acid negatively stained preparations, intact virus particles, 400-450 A.U. total diameter, were observed inside infected cells. In particles in which the peripheral layer became discontinuous, geometrically arranged subunits compatible with cubic symmetry were observed.


[An update on treatment of dementia and Alzheimer's diseases?]
Major and minor neurocognitive disorders are common and increase with age. In the 70s, similarities between " senile " and " presenile " dementia led to their homogenization in a unique illness, the Alzheimer's disease. Systematic specific psychosocial support of patients and caregivers is crucial and allows to discuss an advance care plan. We propose such a multidisciplinary program and discuss some of them. Various drugs and a variety of possible physiopathogenic targets are actually studied and should in the near future offer new therapeutics in dementia and Alzheimer's disease.


Skeletal abnormalities in fetuses with Down's syndrome: a radiographic post-mortem study.
To evaluate skeletal abnormalities on post-mortem radiographs of fetuses with Down's syndrome. Biometrical and morphological criteria, which are used for US prenatal detection of trisomy 21, were assessed. Limb long bones, biparietal diameter (BPD)/occipito-frontal diameter (OFD) ratio, ossification of nasal bones and appearance of the middle phalanx of the fifth digit (P2) in 60 fetuses with Down's syndrome were analysed and compared with 82 normal fetuses matched for gestational age (GA) from 15 to 40 weeks' gestation (WG). We observed reduced growth velocity of limb long bones during the third trimester in both groups, but the reduction was more pronounced in the trisomic group. Brachycephaly was found as early as 15 WG in Down's syndrome and continued throughout gestation (sensitivity 0.28, specificity 1). Ossification of the nasal bones, which can be detected in normal fetuses from 14 WG, was absent in one quarter of trisomic fetuses, regardless of GA. The middle phalanx of the fifth digit was evaluated by comparison with the distal phalanx (P3) of the same digit. We found that P2 was not ossified in 11/31 trisomic fetuses before 23 WG, and was either not ossified or hypoplastic in 17/29 cases after 24 WG (sensitivity 0.56, specificity 1). Three key skeletal signs were present in trisomic fetuses: brachycephaly, absence of nasal bone ossification, and hypoplasia of the middle phalanx of the fifth digit. All these signs are appropriate to prenatal US screening. When present, they fully justify determination of the fetal karyotype by amniocentesis.


Extracellular CADM1 interactions influence insulin secretion by rat and human islet β-cells and promote clustering of syntaxin-1.
Contact between β-cells is necessary for their normal function. Identification of the proteins mediating the effects of β-cell-to-β-cell contact is a necessary step toward gaining a full understanding of the determinants of β-cell function and insulin secretion. The secretory machinery of the β-cells is nearly identical to that of central nervous system (CNS) synapses, and we hypothesize that the transcellular protein interactions that drive maturation of the two secretory machineries upon contact of one cell (or neural process) with another are also highly similar. Two such transcellular interactions, important for both synaptic and β-cell function, have been identified: EphA/ephrin-A and neuroligin/neurexin. Here, we tested the role of another synaptic cleft protein, CADM1, in insulinoma cells and in rat and human islet β-cells. We found that CADM1 is a predominant CADM isoform in β-cells. In INS-1 cells and primary β-cells, CADM1 constrains insulin secretion, and its expression decreases after prolonged glucose stimulation. Using a coculture model, we found that CADM1 also influences insulin secretion in a transcellular manner. We asked whether extracellular CADM1 interactions exert their influence via the same mechanisms by which they influence neurotransmitter exocytosis. Our results suggest that, as in the CNS, CADM1 interactions drive exocytic site assembly and promote actin network formation. These results support the broader hypothesis that the effects of cell-cell contact on β-cell maturation and function are mediated by the same extracellular protein interactions that drive the formation of the presynaptic exocytic machinery. These interactions may be therapeutic targets for reversing β-cell dysfunction in diabetes.


Building bridges between populations and samples in epidemiological studies.
The increased use of rigorous population-sampling methods and the analysis of data from those samples in cross-sectional surveys, case-control studies, longitudinal-cohort investigations, and other epidemiological research efforts have raised important statistical issues for health analysts. We describe the origin, implications, and some plausible resolutions for several of these issues. Some of the main issues we consider include (a) establishing whom the sample represents; (b) using sample weights; (c) understanding the role of other important features, such as the use of sampling stratification and the selection of clustered groups of population members; and (d) finding ways to analyze study data with key sampling features in mind. Ultimately, resolution of all of these issues requires that analysts clearly define a reference population and then understand the role of design features in relating sample results to that population.


Medrysone promotes corneal injury repair by promoting M2-like polarization of macrophages.
Accumulated evidence suggests that M2-like polarized macrophages plays an important role in reducing inflammation, promoting and accelerating wound healing process and tissue repair. Thus, M2-like TAMs (Tumour-associated macrophages) was an appealing target for therapy intervention. Flow cytometry and RT-PCR assay were used to detect the polarization of macrophages induced by Medrysone, and the rat corneal mechanical injury model was established to evaluate the efficacy of Medrysone in cornel repair. Here we found that Medrysone enhanced IL-4 induced M2 polarization of macrophages, as illustrated by increased expression of CD206, up-regulation of M2 marker mRNAs. Medrysone promoted VEGF and CCL2 secretion in IL-4 induced M2-like polarization. IL-4 triggered STAT6 activation was further enhanced by Medrysone and silencing of STAT6 partially abrogated the stimulatory effect of Medrysone. Medrysone improved migration-promoting feature of M2-like macrophages, as indicated by increased migration of endothelial cells. Further, Medrysone promoted corneal injury repair by inducing M2 polarization of macrophages in vivo. Our study suggest that Medrysone promotes corneal injury repair by inducing the M2 polarization of macrophages, providing a theoretical basis for the application of Medrysone in the treatment of corneal injury.


[Campylobacter in stool of patients with irritable bowel syndrome].
According to recent studies prevalence of IBS among patients suffered from of acute enteric infection (AEI) reaches 30%, and 25% of them were accounted to Campylobacter infection. We studied the prevalence of Campylobacter in a feces of Russian IBS patients. Fecal samples received from 39 patients with different types of IBS were studied. Campylobacter were detected by three different methods: culture, real time PCR with the C. jejuni-specific primers and ELISA. Only 7 patients from 39 reported about AEI in the past. C. jejuni in a stool was found in 2 (5%) patients: in the first patient by all three methods and in second one by PCR only. The first patient noted that she suffered from AEI 30 years ago. After two weeks confirmatory testing was positive only in the first patients, in the second one the tests were negative. C. jejuni in a stool was detected in 5% of patients with IBS. Further studies of a microbiota of an intestine in IBS patients using the combination of reliable microbiological and molecular techniques are badly needed.


Cell cycle-dependent dynamics and regulation of mitotic kinesins in Drosophila S2 cells.
Constructing a mitotic spindle requires the coordinated actions of several kinesin motor proteins. Here, we have visualized the dynamics of five green fluorescent protein (GFP)-tagged mitotic kinesins (class 5, 6, 8, 13, and 14) in live Drosophila Schneider cell line (S2), after first demonstrating that the GFP-tag does not interfere with the mitotic functions of these kinesins using an RNA interference (RNAi)-based rescue strategy. Class 8 (Klp67A) and class 14 (Ncd) kinesin are sequestered in an active form in the nucleus during interphase and engage their microtubule targets upon nuclear envelope breakdown (NEB). Relocalization of Klp67A to the cytoplasm using a nuclear export signal resulted in the disassembly of the interphase microtubule array, providing support for the hypothesis that this kinesin class possesses microtubule-destabilizing activity. The interactions of Kinesin-5 (Klp61F) and -6 (Pavarotti) with microtubules, on the other hand, are activated and inactivated by Cdc2 phosphorylation, respectively, as shown by examining localization after mutating Cdc2 consensus sites. The actions of microtubule-destabilizing kinesins (class 8 and 13 [Klp10A]) seem to be controlled by cell cycle-dependent changes in their localizations. Klp10A, concentrated on microtubule plus ends in interphase and prophase, relocalizes to centromeres and spindle poles upon NEB and remains at these sites throughout anaphase. Consistent with this localization, RNAi analysis showed that this kinesin contributes to chromosome-to-pole movement during anaphase A. Klp67A also becomes kinetochore associated upon NEB, but the majority of the population relocalizes to the central spindle by the timing of anaphase A onset, consistent with our RNAi result showing no effect of depleting this motor on anaphase A. These results reveal a diverse spectrum of regulatory mechanisms for controlling the localization and function of five mitotic kinesins at different stages of the cell cycle.


Transcanal tympanoplasty: a 15-year report.
The data presented would seem to support the following conclusions: 1. Transcanal tympanoplasty employing connective tissue underlay grafts, ossicular bone prostheses, and avoidance of mastoid exposure, unless irreversible disease is present, seems to meet contemporary standards of efficacy, safety, and long-term stability when performed on patients with either chronic suppurative otitis or atelectatic otitis. 2. Although chronic suppurative otitis and atelectatic otitis have differing characteristics and are of seemingly different pathogenesis, no statistical differences were found in the pattern of ossicular destruction or in their response to surgical intervation. Hearing results, long-term stability of results, and pattern of complications had no significant differences. 3. Hearing results in those patients with destruction of the malleus handle in whom an L-shaped prosthesis was used were significantly different and poorer than when the malleus was present. A different approach, perhaps employing homograft tympanic membrane with incorporated malleus and staging, may be indicated for these patients. 4. The use of polymeric silicone film and homograft nasal cartilage was associated with a significant number of complications and has been abandoned, substituting instead absorbable gelatin film and ossicular bone transplants. 5. Since no significant changes were seen following one year's observation, this period would seem to be statistically suitable for analysis of results. This conclusion can only be applied to those patients with either chronic suppurative otitis or atelectatic otitis and in whom the surgical procedures herein described were applied.


Potential correlation between EDN1 gene polymorphisms with preeclampsia.
To explore the potential correlation between endothelin 1 (EDN1) gene polymorphisms with preeclampsia (PE). The single nucleotide polymorphisms (SNPs) of 248 PE patients and 232 healthy controls were genotyped by Polymerase Chain Reaction (PCR). The possible association between EDN1 polymorphisms and PE was revealed through the t-test and the Chi-square test. PE risk was significantly correlated with the C allele of polymorphism rs5370 in EDN1. The polymorphism rs5370 in EDN1 was remarkably associated with the onset of severe PE, rather than mild PE. The markedly increased risk of early-onset PE was related to the C allele of polymorphism rs5370 in EDN1, while no significant difference in the allele frequency of polymorphism rs1800541 was detected between the PE group and the control group. In the co-dominant model, the CC genotype of polymorphism rs5370 in EDN1 was associated with the increased PE risk. PE risk in the population carrying TC genotype was 1.59 times higher than those with TT/CC genotype, while polymorphism rs1800541 had no apparent association with PE risk. In the severe PE group, there was an evident difference in the genotype frequency between the dominant and over-dominant models of polymorphism rs5370. In the recessive model, the raised risk of early-onset PE was notably correlated with the TT/CC genotype compared with that of TT genotype. However, no evident association with the genotype frequency of polymorphism rs1800541 was observed between PE patients and controls. EDN1 gene polymorphism rs5370 is correlated with the increased risk of PE.


Site-specific proteoglycan characteristics of third carpal articular cartilage in exercised and nonexercised horses.
The relevance of site and the influence of exercise on third carpal articular cartilage proteoglycan (PG) were assessed in 16 horses. Six horses were exercised (exercised group) for 30 minutes, 3 times/wk, for 6 weeks. The other 10 horses (nonexercised group) were housed in box stalls for the same 6-week period. At week 6, articular cartilage from the proximal surface of the right third carpal bone was harvested and cultured with radioactive sulfate to label newly synthesized PG. Endogenous PG was measured by use of a uronic acid assay. Newly synthesized and endogenous PG were characterized by use of Sepharose CL-2B chromatography, composite gel electrophoresis, and/or immunoblot analysis with monoclonal antibody 1C6 directed against the hyaluronic acid-binding region on PG. There was a significant (P = 0.0002) effect of exercise, but not site, on newly synthesized PG, which was increased in the exercised horses, compared with the nonexercised horses at the end of the 6-week study period. The increase in newly synthesized PG was not reflected in the existing cartilage matrix as there was no significant difference between groups in endogenous PG. However, there was a significant (P = 0.01) effect of site on endogenous PG, with the nest of sites located in the palmar aspect of the radial facet containing a greater concentration of endogenous PG than the nests of sites located on the dorsal aspect of the radial facet or all sites on the intermediate facet. Most newly synthesized PG in both groups consisted of hydrodynamically small PG monomers. However, there was a change in the profile of newly synthesized PG at some sites in the exercised horses to include an early elution peak on Sepharose CL-2B, which may contain aggregating PG. All sites in both groups contained a diverse population of endogenous large and small PG on toluidine blue-stained composite gels that reacted with monoclonal antibody 1C6, indicating the potential to bind to hyaluronic acid.


Examining the links between therapeutic jurisprudence and mental health court completion.
Research demonstrates that mental health courts (MHCs) lead to improved outcomes compared to traditional criminal court processes. An underlying premise of MHCs is therapeutic jurisprudence (TJ). However, no research, to our knowledge, has examined whether MHC outcomes are predicted by TJ principles as theorized. In the present study, we examined whether principles measured at the onset of MHC enrollment (knowledge, perceived voluntariness, and procedural justice) predicted MHC completion (graduation). Using structural equation modeling with MHC participants from four courts, a significant, direct relationship between TJ and MHC completion was found, such that higher levels of TJ were associated with higher rates of success. Although this direct effect became nonsignificant when mediator variables were included, a significant indirect path remained, such that increased levels of initial perceived voluntariness and procedural justice, and MHC knowledge, led to decreased rates of new arrests, prison, MHC bench warrants, and increased court compliance, which, in turn, led to a higher likelihood of MHC graduation.


Screen for low-arsenic-risk rice varieties based on environment-genotype interactions by using GGE analysis.
Arsenic (As) accumulation in rice is a global health concern that has received increased attention in recent years. In this study, 12 rice genotypes were cultivated at four As-contaminated paddy sites in Taiwan. According to the different crop seasons and As levels in the soil, the sites were further divided into 18 environmental conditions. For As in soils, results showed that 67% of the studied environments were likely to represent As contamination. For As in rice, the mean total As concentration in brown rice grains ranged from 0.17 to 0.45 mg kg-1. The analysis of variance for the environment effect indicated that grain As concentration was mainly affected by the environmental conditions, suggesting that there was a remarkable degree of variation across the trial environments. According to the combination of the GGE biplot and cumulative distribution function of order statistics (CDFOS) analysis, five genotypes-TCS17, TCS10, TT30, KH139, and TC192-were regarded as stable, low-risk genotypes because the probability of grain As concentration exceeding the maximum permissible concentration (MPC) was lower for these genotypes across all environmental conditions. Particularly, TCS17 was recommended to be the safest rice genotype. Thus, grain As levels in the selected genotypes were applied to assess the health risk to Taiwanese residents associated with As exposure through rice consumption. Results showed that the upper 75th percentile values of the hazard quotient were all less than unity. This suggested that the health risk associated with consuming the selected rice genotypes was acceptable for most of the residents. The methodology developed here would be applicable to screen for stable, low-As-risk rice genotypes across multiple field environments in other regions or countries.


The Evolving Cystic Fibrosis Microbiome: A Comparative Cohort Study Spanning 16 Years.
The cystic fibrosis (CF) airways are infected with a diverse polymicrobial community. Understanding how changes in the CF microbiome have occurred over time, similar to the observed changes in the prevalence of cultured pathogens, is key in understanding the microbiome's role in disease. Drawing from a prospectively collected and maintained sputum biobank, we identified 45 patients with sputum samples collected between the ages of 18 and 21 years in three successive cohorts of adults transitioning to our CF clinic: A (1997-2000), B (2004-2007), and C (2010-2013). Patient demographics, clinical status, and medications were collected from detailed chart review. Microbial communities were assessed by Ilumina MiSeq sequencing of the variable 3 (V3) region of the 16S rDNA. The three cohorts were similar with respect to baseline demographics. There was a trend toward improved health and use of disease-modifying therapies in each successive cohort. Shannon diversity increased in the most recent cohort, suggesting an increase in the diversity of organisms between cohorts. Furthermore, the proportion of samples with Pseudomonas-dominated communities decreased over time, whereas Streptococcus increased. Although β-diversity was associated with transition cohort, the greatest predictor of diversity remained lung function. Furthermore, core microbiome constituents were preserved across cohorts. Modest changes in the composition and structure of the microbiome of three successive cohorts of young adults with CF were observed, occurring in parallel with successive improvements in clinical status. Importantly, however, the core microbiome constituents were preserved across cohorts.


Dyskinesias elicited by methamphetamine: susceptibility of former methadone-consuming monkeys.
Rhesus monkeys with a history of drinking methadone but currently drug-free and control monkeys with no drug history were injected with methamphetamine hydrochloride (2 to 5 milligrams per kilogram of body weight). In six of seven monkeys which had consumed methadone the lowest dose immediately elicited pronounced oral dyskinesias virtually identical to those of human tardive dyskinesia. The control monkeys did not exhibit oral dyskinesias even after prolonged treatment with the highest dose. The clincial implications may be related to the functioning of brain dopaminergic systems.


[Thermoregulation behavior in necrophagous dipteran larvae].
The forensic entomology is the use of insects to date the death. The forensic expert assessment is based on the development of necrophagous insects which are growing on the cadaver, to calculate their age and then estimate the Post-Mortem Interval. This development depends on a number of parameters like temperature, species or behavior. The French Forensic Taphonomy unit, the only expert team on the subject in France, works on the biology, physiology and ethology of the necrophagous insects. Their works are focused on thermoregulation behavior and thermal preferendum of maggot masses, aggregation phenomenon and social interaction or on food intake. These works are particularly of interest to understand the pre-social parameters and evolution strategies. More importantly, their aim is to better understand the development of necrophagous insects and, in fine, to improve the forensic expert assessment.


[Whole genome discovery of genes related to branching and co-localization with QTLs in soybean].
Branch number of soybean is one of agronomic traits that closely related with yield in both individual and population levels; therefore, discovering genes related to branching is important for molecular breeding in soybean. In this study, 183 genes related to plant branch development had been collected through GO classification and literatures. Based on the principle of similar sequences and conserved domains, 406 genes possibly related to branching had been identified at the whole genome level in soybean. On the other hand, integration of published QTLs related to branching was carried out using BioMercator2.1, and a total of 35 QTLs was located on 12 chromosomes. Co-localization analysis of QTLs and genes related to branching suggested that 57 genes were located in 20 intergrated QTL regions. These results provided useful in-formation for fine mapping of QTLs related to branching and molecular biology study of branch development in soybean.


Evaluation of a drift flux model for simulating submicrometer aerosol dynamics in human upper tracheobronchial airways.
In this study, a hybrid drift flux velocity correction (DF-VC) model that accounts for both submicrometer particle diffusion and inertia was extended to transient conditions and was tested against existing experimental deposition data measured in a replica cast of the human tracheobronchial (TB) region for laminar and turbulent flow. To evaluate the effectiveness of the DF-VC model, deposition results were compared with a standard chemical species (CS) approach that neglects particle inertia. A numerical model of the TB cast was constructed from CT images and extended from the larynx to approximately the sixth respiratory generation. Experimentally determined inlet and outlet flow conditions were implemented in the computational model to ensure direct comparisons between simulations and measurements for the deposition of 40 and 200 nm particles. A low Reynolds number k-omega turbulence model was employed to resolve the laminar and turbulent flow regimes that coexist in the TB geometry. Interesting flow characteristics were observed due to the presence of the larynx, asymmetrical ventilation, and left-right asymmetry, which created a right-skewed laryngeal jet and flow reversal in the trachea that persist over a majority of the transient flow cycle. In comparison with the CS model, deposition results of the DF-VC approach persistently agreed better with experimental findings on a total and sub-branch basis, which indicated that the DF-VC model effectively captured the influence of finite particle inertia. For the submicrometer aerosols considered, transient flows were observed to increase deposition arising from impaction and decrease deposition arising from diffusion on a total and segmental basis compared with steady state conditions. However, the maximum deposition enhancement factor was significantly increased under transient conditions for both 40 nm (factor of 2) and 200 nm (factor of 7) aerosols. Results of this study indicate that a drift flux particle transport model with near-wall velocity corrections can provide an effective continuous-field approach for simulating the transport and deposition of submicrometer respiratory aerosols in human upper TB airways.


[Studies on lysine availability with Tetrahymena pyriformis as the test organism].
A series of experiments was performed to verify the suitability of the protozoon Tetrahymena pyriformis (T.P.) for determining the lysine availability of feeds. The multiplication rate was estimated with the aid of organism counting, turbidimetry and reduction of triphenyltetrazolium chloride. The volume of T.P. was calculated on the basis of length and breadth measurements, Within higher rates of multiplication, there was connected a tendency towards larger organism volumes. A comparison of the growth tests showed that only turbidimetry is able to reflect, under certain conditions, the efficiency of the test organism in protein synthesis. The values for lysine availability varied widely according to the kind and pretreatment of the feedstuff, but did in no case attain those obtained with animal experiments. T.P. is, therefore, scarcely suited as a test organism for determining the protein quality and amino-acid availability of feedstuffs.


Clinical significance of concurrent hepatitis B surface antigen and antibody positivity.
Over a period of 30 months, 64 patients with concurrently positive hepatitis B surface antigen (HBsAg) and antibody (anti-HBs) were identified at two institutions. When all assays were considered, 23.9% of HBsAg-positive individuals exhibited anti-HBs. Both persistent HBsAg/anti-HBs positivity and variable changes in antigen or antibody status were observed among the 36 patients with follow-up beyond six months. When compared to a control group, these patients did not exhibit differences in risk factors for acquiring hepatitis B or in clinical and histological diagnoses. Concurrent HBsAg and anti-HBs was present at detection in 25 patients, while antibody appeared in the remaining 11 subjects. Subsequently, it was undetectable in seven of these 36 patients; no clinical changes occurred at the time of acquisition or loss of the antibody. The titer of the antibody was below 10 mIU/ml in 75% of these individuals. In 10 patients, the subtype of HBsAg was ad, while the anti-HBs was anti-y. Concurrent HBsAg and anti-HBs is a pattern frequently observed throughout the spectrum of hepatitis B-related events. The heterotypic antibody in these patients is of a low titer, and its appearance or disappearance is not associated with changes in the clinical course; simultaneous HBsAg/anti-HBs positivity does not appear to reflect a distinct clinical entity.


Genetic dissection of the planarian reproductive system through characterization of Schmidtea mediterranea CPEB homologs.
Cytoplasmic polyadenylation is a mechanism of mRNA regulation prevalent in metazoan germ cells; it is largely dependent on Cytoplasmic Polyadenylation Element Binding proteins (CPEBs). Two CPEB homologs were identified in the planarian Schmidtea mediterranea. Smed-CPEB1 is expressed in ovaries and yolk glands of sexually mature planarians, and required for oocyte and yolk gland development. In contrast, Smed-CPEB2 is expressed in the testes and the central nervous system; its function is required for spermatogenesis as well as non-autonomously for development of ovaries and accessory reproductive organs. Transcriptome analysis of CPEB knockdown animals uncovered a comprehensive collection of molecular markers for reproductive structures in S. mediterranea, including ovaries, testes, yolk glands, and the copulatory apparatus. Analysis by RNA interference revealed contributions for a dozen of these genes during oogenesis, spermatogenesis, or capsule formation. We also present evidence suggesting that Smed-CPEB2 promotes translation of Neuropeptide Y-8, a prohormone required for planarian sexual maturation. These findings provide mechanistic insight into potentially conserved processes of germ cell development, as well as events involved in capsule deposition by flatworms.


[Characteristics of the nursing instructor which facilitate the process of becoming a nurse. Preliminary note].
Our experience as nursing educator allows us to affirm that, as a general rule, there is a constant concern on that part of nursing teachers from the various schools and universities for the formation of the future professionals. Thus, with the purpose of improving the quality of teaching, these educations continually analyze and evaluate their programs, planning and implementing new or different theoretical and practical activities. However, notwithstanding that concern, we have observed that the professional behaviour of most of staff RN's, especially those recent graduated, does not reflect much of what was taught to them in nursing school. Often staff, nurses manifest dissatisfaction with their professional practice, although little or nothing seems to be done by them in order to change their professional reality and obtain self-achievement. Without any doubt, there are countless number of factors intervening and determining the above outlined situation, but, certainly one of them is the quality that these nurses attribute to the different learning situations experienced by them while students in their nursing programs. The person of the nursing instructor is always a relevant component in these situations. It is inquestionable that the nursing teacher, as any teacher, exerts a great influence upon the quality of the learning experiences of his students, as well as upon their lifes. By way of his attitude towards the educational process, particularly with respect to the subject of this process - the student - the teacher may, or may not, contribute in a consistent and significant way to the emotional maturation and to the personal and professional growth of this students, facilitating, therefore, the process by which these students become professionals. The lack of studies done in our midst on these fundamentals aspects of the nursing teaching - learning process, seems to indicate that, in general, nursing teacher confer a greater importance to the quality of teaching thin to the quality of learning. Concerned with these observations in a time which we consider critical for professional nursing in Brazil, we have been intensely motivated to identify the characteristics of the nursing teacher that enriches the student's learning experiences, and, consequently, facilitate to him the process of becoming a nurse.


Nonspecific vs. specific DNA binding free energetics of a transcription factor domain protein.
Transcription factor (TF) proteins regulate gene expression by binding to specific sites on the genome. In the facilitated diffusion model, an optimized search process is achieved by the TF alternating between 3D diffusion in the bulk and 1D diffusion along DNA. While undergoing 1D diffusion, the protein can switch from a search mode for fast diffusion along nonspecific DNA to a recognition mode for stable binding to specific DNA. It was recently noticed that, for a small TF domain protein, reorientations on DNA happen between the nonspecific and specific DNA binding. We here conducted all-atom molecular dynamics simulations with steering forces to reveal the protein-DNA binding free energetics, confirming that the search and recognition modes are distinguished primarily by protein orientations on the DNA. As the binding free energy difference between the specific and nonspecific DNA system slightly deviates from that being estimated directly from dissociation constants on 15-bp DNA constructs, we hypothesize that the discrepancy can come from DNA sequences flanking the 6-bp central binding sites that impact on the dissociation kinetics measurements. The hypothesis is supported by a simplified spherical protein-DNA model along with stochastic simulations and kinetic modeling.


Design principles for bifunctional targeted oligonucleotide enhancers of splicing.
Controlling the patterns of splicing of specific genes is an important goal in the development of new therapies. We have shown that the splicing of a refractory exon, SMN2 exon 7, could be increased in fibroblasts derived from patients with spinal muscular atrophy by using bifunctional targeted oligonucleotide enhancers of splicing (TOES) oligonucleotides that anneal to the exon and contain a 'tail' of enhancer sequences that recruit activating proteins. We show here that there are striking agreements between the effects of oligonucleotides on splicing in vitro and on both splicing and SMN2 protein expression in patient-derived fibroblasts, indicating that the effects on splicing are the major determinant of success. Increased exon inclusion depends on the number, sequence and chemistry of the motifs that bind the activator protein SRSF1, but it is not improved by increasing the strength of annealing to the target site. The optimal oligonucleotide increases protein levels in transfected fibroblasts by a mean value of 2.6-fold (maximum 4.6-fold), and after two rounds of transfection the effect lasted for a month. Oligonucleotides targeted to the upstream exon (exon 6 in SMN) are also effective. We conclude that TOES oligonucleotides are highly effective reagents for restoring the splicing of refractory exons and can act across long introns.


[U.S. Supreme Court ruling on patentability of genes].
This paper involves a critical commentary concerning the judgment of the U.S. Supreme Court 13/06/2013 on the patentability of human genes. In the author's opinion, the judgment reaffirms a favorable position to a current school of thought whereby protecting "biotechnological innovations" distorts the patent law.


Neuropsychological functioning in ageing retired NFL players: a critical review.
Recent discovery of chronic traumatic encephalopathy in former National Football League (NFL) players has led to a surge of papers investigating cognitive functioning in these former athletes. This critical review of the literature focused on the neuropsychological functioning in these ageing athletes, and included 22 articles published between 2013 and 2019, of which 13 reported on neuroradiological imaging and four focused on dose-response relationships of repetitive head injury on cognitive outcomes. Four studies suggest higher prevalence of MCI and neurodegenerative disease among NFL retirees, although a quantifiable risk and prevalence of cognitive impairment and dementia in these players remains unknown. Decreased verbal memory has been found in some players across multiple studies, though with unknown clinical significance due to small sample sizes, unreported effect sizes, and absence of longitudinal data. Studies investigating a dose-response relationship between cognitive decline and head injury have generated mixed findings utilizing various measures of head injury exposure. Neuroradiological findings are inconsistent, but suggest that some NFL players may be at greater risk for reduced white matter integrity. Future research is needed to understand the relationship between sports-related concussions and the risk of long-term cognitive decline and neurodegenerative disease in ageing NFL players.


Mesenchymal stem cell transplantation into the spinal cord of healthy adult horses undergoing cervical ventral interbody fusion.
To determine the feasibility of umbilical cord-derived mesenchymal stem cell (UC-MSC) transplantation into the cervical spinal cord of horses by using fluoroscopy with or without endoscopic guidance and to evaluate the neurological signs and tissue reaction after injection. Experimental study. Eight healthy adult horses with no clinical signs of neurological disease. After cervical ventral interbody fusion (CVIF), ten million fluorescently labeled allogeneic UC-MSC were injected into the spinal cord under endoscopic and fluoroscopic guidance (n = 5) or fluoroscopic guidance only (n = 3). Postoperative neurological examinations were performed, and horses were humanely killed 48 hours (n = 4) or 14 days (n = 4) postoperatively. Spinal tissues were examined after gross dissection and with bright field and fluorescent microscopy. Needle endoscopy of the cervical canal by ventral approach was associated with intraoperative spinal cord puncture (2/5) and postoperative ataxia (3/5). No intraoperative complications occurred, and one (1/3) horse developed ataxia with cell transplantation under fluoroscopy alone. Umbilical cord-derived MSC were associated with small vessels and detected up to 14 days in the spinal cord. Demyelination was observed in six of eight cases. Fluoroscopically guided intramedullary UC-MSC transplantation during CVIF avoids spinal cord trauma and decreases risk of ataxia from endoscopy. Umbilical cord-derived MSC persist in the spinal cord for up to 14 days. Cell injection promotes angiogenesis and induces demyelination of the spinal tissue. Umbilical cord-derived MSC transplantation into the spinal cord during CVIF without endoscopy is recommended for future evaluation of cell therapy in horses affected by cervical vertebral compressive myelopathy.


In vitro antimicrobial activity of cefotaxime, a new cephalosporin.
Cefotaxime, a new semisynthetic cephalosporin derivative, showed a broad spectrum of antibacterial activity against clinically isolated strains of gram-positive and gram-negative bacteria. This cephalosporin was slightly less active than cefazolin against Staphylococcus aureus but 4 to 300 times as active as carbenicillin against gram-negative organisms, including Pseudomonas aeruginosa, Pseudomonas cepacia, Enterobacter cloacae, and Serratia marcescens. Cefotaxime was the most active compound against members of the Enterobacteriaceae and 20- to 100-fold more active than cefoxitin against the indole-positive Proteus group. The minimal bactericidal concentrations of the compound were identical to, or two times higher than, the minimal inhibitory concentrations against Escherichia coli and P. aeruginosa and four times higher against S. marcescens. A reduction of inoculum size decreased greatly the minimal inhibitory and bactericidal concentrations of cefotaxime against E. coli P. aeruginosa, and S. marcescens. The antibiotic was very stable to penicillinase and cephalosporinase produced by gram-negative bacteria, including Proteus vulgaris.


Elevated Hypothalamic Glucocorticoid Levels Are Associated With Obesity and Hyperphagia in Male Mice.
Glucocorticoid (Gc) excess, from endogenous overproduction in disorders of the hypothalamic-pituitary-adrenal axis or exogenous medical therapy, is recognized to cause adverse metabolic side effects. The Gc receptor (GR) is widely expressed throughout the body, including brain regions such as the hypothalamus. However, the extent to which chronic Gcs affect Gc concentrations in the hypothalamus and impact on GR and target genes is unknown. To investigate this, we used a murine model of corticosterone (Cort)-induced obesity and analyzed Cort levels in the hypothalamus and expression of genes relevant to Gc action. Mice were administered Cort (75 μg/mL) or ethanol (1%, vehicle) in drinking water for 4 weeks. Cort-treated mice had increased body weight, food intake, and adiposity. As expected, Cort increased plasma Cort levels at both zeitgeber time 1 and zeitgeber time 13, ablating the diurnal rhythm. Liquid chromatography dual tandem mass spectrometry revealed a 4-fold increase in hypothalamic Cort, which correlated with circulating levels and concentrations of Cort in other brain regions. This occurred despite decreased 11β-hydroxysteroid dehydrogenase (Hsd11b1) expression, the gene encoding the enzyme that regenerates active Gcs, whereas efflux transporter Abcb1 mRNA was unaltered. In addition, although Cort decreased hypothalamic GR (Nr3c1) expression 2-fold, the Gc-induced leucine zipper (Tsc22d3) mRNA increased, which indicated elevated GR activation. In keeping with the development of hyperphagia and obesity, Cort increased Agrp, but there were no changes in Pomc, Npy, or Cart mRNA in the hypothalamus. In summary, chronic Cort treatment causes chronic increases in hypothalamic Cort levels and a persistent elevation in Agrp, a mediator in the development of metabolic disturbances.


How does cognitive reappraisal affect the response to gains and losses?
To investigate the influence of cognitive reappraisal, one important kind of emotion regulation strategy, on psychological and electrophysiological responses to gains and losses, a monetary gambling task was performed in two conditions, that is, spontaneity and regulation. Event-related potentials (ERP) and self-rating emotional experiences to outcome feedback were recorded during the task. Cognitive reappraisal reduced self-rating emotional experience to both gains and losses and the amplitudes of the feedback-related negativity (FRN) and the P3 of ERPs. According to these results, we suggest that the application of cognitive reappraisal strategy significantly modulated the motivational salience of current outcomes, thus weakening the subjective emotional experience elicited. In addition, cognitive reappraisal might have changed the allocation of cognitive resources during outcome evaluation. This study extends emotion regulation studies by applying monetary outcomes as emotional stimuli, and also implicates the significance of emotion regulation in decision-making processes.


Identification and mapping of linear antibody epitopes in human serum albumin using high-density Peptide arrays.
We have recently developed a high-density photolithographic, peptide array technology with a theoretical upper limit of 2 million different peptides per array of 2 cm(2). Here, we have used this to perform complete and exhaustive analyses of linear B cell epitopes of a medium sized protein target using human serum albumin (HSA) as an example. All possible overlapping 15-mers from HSA were synthesized and probed with a commercially available polyclonal rabbit anti-HSA antibody preparation. To allow for identification of even the weakest epitopes and at the same time perform a detailed characterization of key residues involved in antibody binding, the array also included complete single substitution scans (i.e. including each of the 20 common amino acids) at each position of each 15-mer peptide. As specificity controls, all possible 15-mer peptides from bovine serum albumin (BSA) and from rabbit serum albumin (RSA) were included as well. The resulting layout contained more than 200.000 peptide fields and could be synthesized in a single array on a microscope slide. More than 20 linear epitope candidates were identified and characterized at high resolution i.e. identifying which amino acids in which positions were needed, or not needed, for antibody interaction. As expected, moderate cross-reaction with some peptides in BSA was identified whereas no cross-reaction was observed with peptides from RSA. We conclude that high-density peptide microarrays are a very powerful methodology to identify and characterize linear antibody epitopes, and should advance detailed description of individual specificities at the single antibody level as well as serologic analysis at the proteome-wide level.


Behavior of venous thrombi: historical observations.
Charles White in 1784 first demonstrated that milk leg was not caused by retained milk or lochia but rather by obstructing clots in the veins. In 1847, Virchow observed that venous thrombi often migrated to the lungs. Workers since then have found that, in general, thrombi in veins and in pulmonary arteries tend to form large recanalizing channels, accompanied by the destruction of any valves that are present, but that those in arteries form only small "arteries of organization." On the other hand, observers have reported capricious instances of opposite behavior. Furthermore, fresh thrombi and emboli in both systems occasionally disappear in the early period after formation or lodgement. It is urged that we use the full potential of modern diagnostic and monitoring methods to procure more data on the details of intravascular clot behavior.


Postoperative myocardial ischemic events.
During the last 15 years, improved intraoperative hemodynamic management has helped reduce the incidence of myocardial ischemic events, and most events now occur outside the operating room in the first 72 hours following surgery. Currently, what is considered cost-effective management monitors high risk patients in the intensive care unit postoperatively, before transferring them to an unmonitored nursing unit. However, in both monitored and unmonitored areas ischemia is usually not appreciated unless clinical symptoms are present. Thus, our current system of delivering postoperative care is both inefficient and costly. It is clear that we are on the verge of a new frontier in our ability to detect and diagnose perioperative myocardial ischemic events. The possibility of acute real-time intervention and accurate risk stratification using surgery as a stress test is exciting. The challenge at hand is to utilize these modalities in the most beneficial, cost-effective manner. Only through continued quality research can this goal be attained.


DNA synapsis through transient tetramerization triggers cleavage by Ecl18kI restriction enzyme.
To cut DNA at their target sites, restriction enzymes assemble into different oligomeric structures. The Ecl18kI endonuclease in the crystal is arranged as a tetramer made of two dimers each bound to a DNA copy. However, free in solution Ecl18kI is a dimer. To find out whether the Ecl18kI dimer or tetramer represents the functionally important assembly, we generated mutants aimed at disrupting the putative dimer-dimer interface and analysed the functional properties of Ecl18kI and mutant variants. We show by atomic force microscopy that on two-site DNA, Ecl18kI loops out an intervening DNA fragment and forms a tetramer. Using the tethered particle motion technique, we demonstrate that in solution DNA looping is highly dynamic and involves a transient interaction between the two DNA-bound dimers. Furthermore, we show that Ecl18kI cleaves DNA in the synaptic complex much faster than when acting on a single recognition site. Contrary to Ecl18kI, the tetramerization interface mutant R174A binds DNA as a dimer, shows no DNA looping and is virtually inactive. We conclude that Ecl18kI follows the association model for the synaptic complex assembly in which it binds to the target site as a dimer and then associates into a transient tetrameric form to accomplish the cleavage reaction.