Visualization and characterization of gastric contractions using a radionuclide technique.
With the use of the radionuclide gastric-emptying test and a new data processing method, the contraction characteristics of the stomach were analyzed. After ingestion of a radiolabeled test meal, dynamic images of the stomach were acquired and analyzed to determine the frequency, amplitude, and rate of gastric contractions in healthy subjects. The frequency of antral contractions was found to be inversely related with food retention in the stomach; in contrast, the amplitude of the contractions decreased progressively during the course of gastric emptying. The peaks of both antral contraction and filling rate and the time of their occurrence remained constant throughout gastric emptying. The observed patterns of phase distribution and sequential phase changes of the food in the stomach confirmed noninvasively what was already known from invasive technique, i.e., that the proximal stomach does not undergo phasic contractions and that, in the distal stomach, smooth muscle contraction originates in midcorpus and propagates aborally to the pylorus. The scintigraphic test can be used to noninvasively and quantitatively characterize gastric motor function and to delineate the spatial sequence of gastric contractions. This technique can be applied to study the pathophysiology of gastric emptying in various motor disorders.


Application of Raw and Defatted by Supercritical CO2 Hemp Seed Press-Cake and Sweet Grass Antioxidant Extract in Pork Burger Patties.
There is an increasing tendency toward the application of plant origin ingredients in meat products. This study evaluates the physicochemical properties and oxidative stability of pork burger patties produced with the addition of dried raw and defatted by supercritical CO2 extraction hemp seed press-cake as protein-rich ingredients (1.5-2.0%) and sweet grass ethanolic extract (0.5%) as a strong natural antioxidant. The main aim of using such a combination was to assess the possibility of mitigating the negative effects of hemp seed press-cake, containing approx. 10% of highly unsaturated oil, on the oxidation of meat products. The patties were compared with the control sample (without additives) during storage on days 0, 4, 8, 15, and 21 at 4 °C in modified atmosphere conditions. Plant ingredients reduced the lightness of pork patties, while their effects on other physicochemical characteristics were insignificant. The patties with fully defatted hemp seed flour showed the lowest grilling losses. Based on the measurement of thiobarbituric acid reactive substances, raw hemp seed press-cake increased the oxidative rate of pork patties; however, remarkably, the addition of sweet grass extract completely inhibited oxidation during the whole period of storage. The sensory characteristics of the products were acceptable; however, the patties with sweet grass extract received lower evaluation scores.


Educational level and risk for alcohol abuse and dependence: differences by race-ethnicity.
Using prospective data, we extend and strengthen prior evidence on the relationship of educational level and the risk of alcohol abuse and dependence among adults. Hypothesizing that risk of alcohol disorders would be greater among individuals who dropped out of high school without getting a diploma, and among those who entered college but then failed to get a degree, relative to those with a college or higher degree, we examined these relationships by race-ethnicity. Data are from the total of 18,571 adult participants selected for the Epidemiologic Catchment Area (ECA) program by probability sampling of households and census tracts between 1980 and 1984. To assess occurrence of psychiatric conditions over time, staff administered the Diagnostic Interview Schedule soon after sampling and again at followup approximately one year later. The final sample for this report was stratified: African Americans (N = 2856) and whites (N = 7889). In multiple logistic regression analyses, we found that dropping out of high school was associated with elevated risk for alcohol disorders, relative to those with an Associate of Arts degree or higher, among both African Americans (estimated relative risk [RR] = 4.0, 95% confidence interval [CI], 1.1-14.3, P = .03), and whites (RR = 4.7, 95% CI, 2.2-10.0, P = .0001). In contrast, entering college but failing to get a degree was associated with a significantly increased risk only for whites; however, African Americans were less numerous in the sample, causing attenuation of power. In order to assess whether this relationship was confounded by macrosocial factors of the neighborhood environment, cases were matched to non-cases by age and census tract, and initial conditional logistic regression analyses also were completed. If confirmed in other investigations, these findings may help to identify groups that are at higher risk for developing alcohol abuse and alcohol dependence in adulthood, and may aid in the development of prevention or early intervention programs.


Long-term survival of an urban fruit bat seropositive for Ebola and Lagos bat viruses.
Ebolaviruses (EBOV) (family Filoviridae) cause viral hemorrhagic fevers in humans and non-human primates when they spill over from their wildlife reservoir hosts with case fatality rates of up to 90%. Fruit bats may act as reservoirs of the Filoviridae. The migratory fruit bat, Eidolon helvum, is common across sub-Saharan Africa and lives in large colonies, often situated in cities. We screened sera from 262 E. helvum using indirect fluorescent tests for antibodies against EBOV subtype Zaire. We detected a seropositive bat from Accra, Ghana, and confirmed this using western blot analysis. The bat was also seropositive for Lagos bat virus, a Lyssavirus, by virus neutralization test. The bat was fitted with a radio transmitter and was last detected in Accra 13 months after release post-sampling, demonstrating long-term survival. Antibodies to filoviruses have not been previously demonstrated in E. helvum. Radio-telemetry data demonstrates long-term survival of an individual bat following exposure to viruses of families that can be highly pathogenic to other mammal species. Because E. helvum typically lives in large urban colonies and is a source of bushmeat in some regions, further studies should determine if this species forms a reservoir for EBOV from which spillover infections into the human population may occur.


[Molecular and cell biological mechanism of olfactory adaptation in Caenorhabditis elegans].
Olfactory adaptation is an important physiological function of animals, which can protect their own neurons from overstimulation, and be better to deal with all kinds of stimuli in the surrounding environment. In this article, we discuss the neuronal basis of olfactory adaptation in Caenorhabditis elegans. Up to now, several intracellular regulatory factors have been discovered to be associated with olfactory adaptation in Caenorhabditis elegans, including cyclic guanosine monophosphate (cGMP) signaling in the olfactory neurons AWC, OSM-9 in transient receptor potential vanilloid (TRPV) channel, arrestin ARR-1, diglyceride (DAG) pathway in G protein signaling pathways, etc. However, the neural circuits of the olfactory adaptation remains largely unknown. This paper reviews molecular and cell biological mechanism of olfactory adaptation in Caenorhabditis elegans, so as to provide reference for studies on olfactory sensation in advanced animals.


Subnormothermic Ex Vivo Porcine Kidney Perfusion Improves Energy Metabolism: Analysis Using 31P Magnetic Resonance Spectroscopic Imaging.
The ideal preservation temperature for donation after circulatory death kidney grafts is unknown. We investigated whether subnormothermic (22 °C) ex vivo kidney machine perfusion could improve kidney metabolism and reduce ischemia-reperfusion injury. To mimic donation after circulatory death procurement, kidneys from 45-kg pigs underwent 60 min of warm ischemia. Kidneys were then perfused ex vivo for 4 h with Belzer machine perfusion solution UW at 22 °C or at 4 °C before transplantation. Magnetic resonance spectroscopic imaging coupled with LCModel fitting was used to assess energy metabolites. Kidney perfusion was evaluated with dynamic-contrast enhanced MRI. Renal biopsies were collected at various time points for histopathologic analysis. Total adenosine triphosphate content was 4 times higher during ex vivo perfusion at 22 °C than at 4 °C perfusion. At 22 °C, adenosine triphosphate levels increased during the first hours of perfusion but declined afterward. Similarly, phosphomonoesters, containing adenosine monophosphate, were increased at 22 °C and then slowly consumed over time. Compared with 4 °C, ex vivo perfusion at 22 °C improved cortical and medullary perfusion. Finally, kidney perfusion at 22 °C reduced histological lesions after transplantation (injury score: 22 °C: 10.5 ± 3.5; 4 °C: 18 ± 2.25 over 30). Ex vivo kidney perfusion at 22°C improved graft metabolism and protected from ischemia-reperfusion injuries upon transplantation. Future clinical studies will need to define the benefits of subnormothermic perfusion in improving kidney graft function and patient's survival.


[Dimethyl sulfoxide in the therapy of vibration disease].
The article contains results of clinical studies of dimethylsulfoxide (DMSO) therapeutic application at vibration disease (VD) cases caused by local vibration and in comparison with the generally accepted complex therapy. 30% DMSO water solution skin compresses were used applied to the affected zones of the upper extremities for 1-1.5 hours daily with 12-15 procedures in the whole course. It was established that DMSO effects are positive in most VB manifestations: regional angiodystonia, sensor and vegetative--sensor polyneuropathy, arthroses, and particularly in cases of muscle dystonia and myodistrophy, humeroscapular periarthrosis. This kind of technique simplifies VD therapy as it excludes physiotherapy procedures and limits medication to a great extent, doing with no drugs at all in some cases. Treatment costs and clinical course duration are also lowered.


Mimotopes of polyreactive anti-DNA antibodies identified using phage-display peptide libraries.
Three monoclonal IgG2a anti-DNA polyreactive autoantibodies, derived from lupus-prone mice (NZB x NZW)F1, were studied by surface plasmon resonance (BIAcore) analysis using three different synthetic double-stranded (ds) oligonucleotides of 25, 30, and 25 base pairs (bp). These monoclonal antibodies (mAb) exhibited dissociation rate constants (k(off)), ranging from 0.0001 (mAb F14.6 and F4.1) to 0.01/s (mAb J20.8) and k(on) ranging from 2 x 10(5) to 2 x 10(6) /M/s. The screening of a constrained random peptide library displayed on M13 bacteriophages on these mAb allowed the determination of the specific consensus motifs (mimotopes) for mAb F14.6 and J20.8, but not for mAb F4.1. No cross-reaction was observed between F14.6- and J20.8-specific peptides (and vice versa). Binding of all phages selected on F14.6 was inhibited with 700 ng/ml soluble DNA. The binding of one group of peptides selected on J20.8 was inhibited by 400 ng/ml soluble DNA, of a second group by 2500 ng/ml, while binding of a third group could not be inhibited. The determined consensus sequences do not match with known sequences. Peptides specific for F14.6 share negative charges and aromatic rings that may mimic a DNA backbone, while peptides selected with J20.8 do not bear any negative charge, implying a different kind of molecular recognition, for example hydrogen or salt bonds. The peptides selected on J20.8 also bind serum antibodies from human patients with systemic lupus erythematosus. In addition, BALB/c mice immunized with some of the selected phages exhibit high serum titers of IgG3 anti-dsDNA antibodies, further supporting the hypothesis that peptide epitopes may mimic an oligonucleotide structure.


Moderate-intensity physical activity, hippocampal volume, and memory in older adults with mild cognitive impairment.
Greater physical activity (PA) is associated with better memory performance and greater hippocampal volume in older adults. However, most studies to date assessed PA via questionnaires and thereby lacked objective characterization of PA (eg, intensity, duration, etc.). Thus, we currently do not have a comprehensive understanding of PA characteristics that are important for neuroprotection, especially among older adults with mild cognitive impairment (MCI). Thus, using triaxial accelerometers, we examined the association between light- and moderate-intensity PA, total duration of PA, hippocampal volume, and memory in older adults with MCI. This cross-sectional study involved 310 older adults with MCI who completed neuropsychological tests of memory, and structural magnetic resonance imaging. Participants were instructed to wear the accelerometer on an elastic band on their hip at all times for 2 weeks. Average daily duration of light, moderate, and total PA (min/day) was calculated. Moderate PA was associated with hippocampal volume (β = .167, p = .003) after controlling for age, but light PA (β = -.021, p = .713) and total PA (β = .011, p = .844) were not. Both light and moderate PAs were not associated with memory performance. Structural equation modeling demonstrated that moderate PA was not directly associated with memory but significantly contributed to hippocampal volume; hippocampal volume loss was significantly and directly associated with poor memory performance. Our results suggest that the benefits of moderate PA on memory among older adults with MCI are mediated by hippocampal volume. Furthermore, light PA may not reduce dementia risk among older adults with MCI.


Inhibition of class IA PI3K enzymes in non-small cell lung cancer cells uncovers functional compensation among isoforms.
Deregulation of the phosphatidylinositol 3-kinase (PI3K) pathway is central to many human malignancies while normal cell proliferation requires pathway functionality. Although inhibitors of the PI3K pathway are in clinical trials or approved for therapy, an understanding of the functional activities of pathway members in specific malignancies is needed. In lung cancers, the PI3K pathway is often aberrantly activated by mutation of genes encoding EGFR, KRAS, and PIK3CA proteins. We sought to understand whether class IA PI3K enzymes represent rational therapeutic targets in cells of non-squamous lung cancers by exploring pharmacological and genetic inhibitors of PI3K enzymes in a non-small cell lung cancer (NSCLC) cell line system. We found that class IA PI3K enzymes were expressed in all cell lines tested, but treatment of NSCLC lines with isoform-selective inhibitors (A66, TGX-221, CAL-101 and IC488743) had little effect on cell proliferation or prolonged inhibition of AKT activity. Inhibitory pharmacokinetic and pharmacodynamic responses were observed using these agents at non-isoform selective concentrations and with the pan-class I (ZSTK474) agent. Response to pharmacological inhibition suggested that PI3K isoforms may functionally compensate for one another thus limiting efficacy of single agent treatment. However, combination of ZSTK474 and an EGFR inhibitor (erlotinib) in NSCLC resistant to each single agent reduced cellular proliferation. These studies uncovered unanticipated cellular responses to PI3K isoform inhibition in NSCLC that does not correlate with PI3K mutations, suggesting that patients bearing tumors with wildtype EGFR and KRAS are unlikely to benefit from inhibitors of single isoforms but may respond to pan-isoform inhibition.


Are Hip Physical Examination Findings Predictive of Future Lower-Body Injury Rates in Elite Adolescent Female Soccer Athletes at Minimum 5-Year Follow-Up?
Although elite adolescent female soccer athletes have unique injury risk factors and management challenges, limited epidemiological data exist for this population. To describe lower-body injury patterns and to determine whether a screening hip physical examination is predictive of future injuries in elite adolescent female soccer athletes. Prospective cohort study. One US premier soccer club. One hundred seventy-seven female soccer athletes aged 10-18 years (mean [SD] 14.6 [1.8] y) completed a demographic questionnaire and screening hip physical examination that included range of motion and provocative tests. At least 5 years after baseline screening, athletes completed an electronic follow-up injury survey. Injury was defined as pain that interfered with sporting activity. In addition to descriptive analyses of athletes' injury profiles, associations between players' baseline demographics and subsequent injury profiles were evaluated using chi-square tests, and potential predictors of injury based on players' baseline hip examinations were evaluated using multivariable logistic regression. Ninety-four of 177 athletes (53%) were contacted for follow-up, and 88/94 (93.6%) completed the survey. With mean follow-up of 91.9 (9.3) months (range 66-108 mo), 42/88 (47.7%) reported sustaining a new lower-body injury. The low back was the most common injury region (16/42, 38.1%). Almost half of all injured athletes (20/42, 47.6%) sustained overuse injuries, and 16/42 (38.1%) had an incomplete recovery. Higher body mass index and reaching menarche were associated with sustaining an injury (P = .03 and .04, respectively). Athletes' baseline hip examinations were not predictive of their subsequent rate of lower-body, lumbopelvic, overuse, or incomplete recovery injury (all P > .05). Lower-body injuries were common in elite adolescent female soccer athletes, with over one third of injured athletes reporting permanent negative impact of the injury on their playing ability. Baseline hip physical examinations were not associated with future injury rate.


The relationship of acromial architecture to rotator cuff disease.
Variations in the architecture of the coracoacromial arch can lead to a clinically symptomatic rotator cuff lesion. Differences in the development and morphology of the acromion, and the presence of anterior acromial spurs and inferior acromioclavicular osteophytes decrease the volume of the subacromial space, leading to impingement. Recent anatomic, radiographic, biomechanical, and SPG studies have confirmed these architectural variations and their effects on the contents of the subacromial space. Abnormal contact between the acromion and these soft tissues can lead to pathologic lesions. Surgical procedures should be directed at increasing the space beneath the coracoacromial arch to reduce wear on the rotator cuff.


Amputation Risk in Patients with Diabetes Mellitus and Peripheral Artery Disease Using Statewide Data.
Conflicting data exist regarding changes in amputation rates in patients with ulcers because of diabetes mellitus (DM) and peripheral artery disease (PAD). This study focuses on how population-based amputation rates are changing in the current treatment era. Using the California Office of Statewide Health Planning and Development Patient Discharge database, all patients who underwent major nontraumatic lower extremity (LE) amputation in 2005 through 2011 were identified. Age-adjusted population-based amputation risk was determined by year. Gender and age trends in amputation risk were estimated separately for diabetes-related amputations and PAD-related amputations, treating all California residents as the population at risk. From 2005 to 2011, 32,025 qualifying amputations were performed in California. Of these, 11,896 were DM-associated (n = 1,095), PAD-associated (n = 4,335), or associated with both conditions (n = 6,466). PAD-associated amputation rates and combined PAD/DM-associated amputation rates have changed little since 2009 after decreasing substantially over the prior 5 years, but DM-associated amputation rates have continuously increased since 2005. California residents older than the age of 80 years had the most dramatic decrease in PAD-associated amputation rates from 2005 to 2011 (i.e., from 317 to 175 per million Californians). Men with PAD/DM had amputation rate 1.5 times higher than those of patients with PAD alone and 5 times higher than rates of DM patients. In women the difference between patient with PAD and PAD/DM was not seen; however, these rates were 2.5 times higher than patients with DM alone. Preventable amputations associated with high-risk diseases are no longer decreasing despite continuing advances in care and education. Octogenarians with PAD represent the highest risk group for amputation, but DM-associated amputations have increased since 2005. Further research to understand treatment pathways for patient with LE wounds may shed light on pathways for amputation prevention in the future.


Chinese Americans' Views and Use of Family Health History: A Qualitative Study.
Family health history (FHH) plays a significant role in early disease detection and prevention. Although Asian Americans are the fastest growing U.S. immigrant group, no data exists regarding Chinese Americans' (the largest Asian subgroup) views and use of FHH. This study examines this important issue. Forty-nine adults from southern U.S. Chinese American communities participated in this qualitative, semi-structured, in-depth interview study. Interviews were audio recorded, transcribed, and analyzed with a content analysis approach. Although the majority of participants perceived the importance of collecting FHH, most lacked FHH knowledge and failed to collect FHH information. Barriers affecting FHH collection and discussion among family members included long-distance separation from family members, self-defined "healthy family," and Chinese cultural beliefs. Lack of doctors' inquiries, never/rarely visiting physicians, self-defined "healthy family," perceived insignificance of discussing FHH with doctors, and Chinese cultural beliefs were the obstacles in communicating FHH with physicians. Chinese Americans had limited usage of their FHH and faced cultural, distance, knowledge-, and healthcare system-related barriers that influenced their FHH use. Developing FHH education programs for Chinese Americans is highly recommended.


Berberine regulates proliferation, collagen synthesis and cytokine secretion of cardiac fibroblasts via AMPK-mTOR-p70S6K signaling pathway.
The traditional Chinese medicinal berberine has long been used to treat cardiovascular diseases; however, the mechanism underlying its effects remains unclear. Here, this study would to investigate the effects of berberine on proliferation, collagen synthesis and cytokine secretion of cardiac fibroblasts. We assessed proliferation, collagen synthesis and cytokine secretion in cardiac fibroblasts subjected to angiotensin II (Ang II) subsequent to the consumption of berberine or a control treatment. And then we detected the role of AMPK/mTOR signaling pathway in berberine treatment of cardiac fibroblasts. In the present study, the cellular behaviors of cardiac fibroblasts induced by Ang II were significantly activated including proliferation, transformation into myofibroblasts and collagen synthesis. Additionally, the ability of cytokine secretion was enhanced obviously. It was demonstrated that treatment of cardiac fibroblasts with berberine resulted in deceased proliferation, and attenuated fibroblast α-smooth muscle actin expression and collagen synthesis. And the protein secretion of TGFβ1 was inhibited; however, the protein secretion of IL-10 was increased in cardiac fibroblasts with berberine treatment. Mechanistically, the phosphorylation level of AMPK was increased; and the phosphorylation levels of mTOR and p70S6K were decreased in berberine treatment group. These results illustrated that the protective effects of berberine on cellular behaviors of cardiac fibroblasts were at least in part due to activate AMPK signaling pathway and downregulate mTOR/p70S6K signaling pathway. Berberine might become a new strategy for treating cardiac fibrosis in the future.


Genomic rearrangements define lineage relationships between adjacent lepidic and invasive components in lung adenocarcinoma.
The development of adenocarcinoma of the lung is believed to proceed from in situ disease (adenocarcinoma in situ, AIS) to minimally invasive disease with prominent lepidic growth (minimally invasive adenocarcinoma, MIA), then to fully invasive adenocarcinoma (AD), but direct evidence for this model has been lacking. Because some lung adenocarcinomas show prominent lepidic growth (AD-L), we designed a study to address the lineage relationship between the lepidic (noninvasive) component (L) and the adjacent nonlepidic growth component representing invasive disease within individual tumors. Lineage relationships were evaluated by next-generation DNA sequencing to define large genomic rearrangements in microdissected tissue specimens collected by laser capture. We found a strong lineage relationship between the majority of adjacent lepidic and invasive components, supporting a putative AIS-AD transition. Notably, many rearrangements were detected in the less aggressive lepidic component, although the invasive component exhibited an overall higher rate of genomic rearrangement. Furthermore, a significant number of genomic rearrangements were present in histologically normal lung adjacent to tumor, but not in host germline DNA, suggesting field defects restricted to zonal regions near a tumor. Our results offer a perspective on the genetic pathogenesis underlying adenocarcinoma development and its clinical management.


Clonality analysis of multifocal papillary thyroid carcinoma by using genetic profiles.
Papillary thyroid carcinoma (PTC) is the most common adult thyroid malignancy and often presents with multiple anatomically distinct foci within the thyroid, known as multifocal papillary thyroid carcinoma (MPTC). The widespread application of the next-generation sequencing technologies in cancer genomics research provides novel insights into determining the clonal relationship between multiple tumours within the same thyroid gland. For eight MPTC patients, we performed whole-exome sequencing and targeted region sequencing to identify the non-synonymous point mutations and gene rearrangements of distinct and spatially separated tumour foci. Among these eight MPTCs, completely discordant mutational spectra were observed in the distinct cancerous nodules of patients MPTC1 and 5, suggesting that these nodules originated from independent precursors. In another three cases (MPTC2, 6, and 8), the distinct MPTC foci of these patients had no other shared mutations except BRAF V600E, also indicating likely independent origins. Two patients (MPTC3 and 4) shared almost identical mutational spectra amongst their separate tumour nodules, suggesting a common clonal origin. MPTC patient 7 had seven cancer foci, of which two foci shared 66.7% of mutations, while the remaining cancer foci displayed no common non-synonymous mutations, indicating that MPTC7 has multiple independent origins accompanied by intraglandular disease dissemination. In this study, we found that 75% of MPTC cases arose as independent tumours, which supports the field cancerization hypothesis describing multiple malignant lesions. MPTC may also arise from intrathyroidal metastases from a single malignant clone, as well as multiple independent origins accompanied by intrathyroidal metastasis.


Evaluation of two novel thoracolumbar trauma classification systems.
Despite numerous attempts at classifying thoracolumbar spinal injuries, there remains no consensus on a single unifying algorithm of management. The ideal system should provide diagnostic and prognostic information, exhibit adequate reliability and validity and be easily applicable to clinical practice. The purpose of this study is to assess the reliability and validity of two novel classification systems for thoracolumbar fractures - the Thoracolumbar Injury Severity Score (TLISS) and the Thoracolumbar Injury Classification and Severity Score (TLICS) - and also to discuss potential efforts towards research in the future. MATEREIALS AND METHODS: Seventy-one patients with thoracolumbar fractures were prospectively assessed by surgeons with different levels of training and experience (attending orthopedic surgeon, attending neurosurgeon, spine fellows, senior level and junior level residents) at a single institution. Plain radiographs, CT and MRI imaging were used to classify these injuries using the TLISS system. Seven months later, 25 consecutive injuries were prospectively assessed with the TLISS and TLICS systems. Unweighted Cohen's kappa coefficients and Spearman's correlation values were calculated to assess inter-observer reliability and validity at each point in time. For both the TLISS and TLICS algorithms, the inter-rater kappa statistics for all of the subgroups demonstrated moderate-to-substantial reliability (0.45-0.74), although there were no significant differences among the shared subgroups. The kappa score of the TLISS system was greater than that of the TLICS system for injury mechanism/ morphology. Correlation values were also greater across all subgroups (P ≤0.01). Statistically significant improvements in TLISS inter-observer reliability were observed across all TLISS fields (P <0.05). The TLISS and TLICS schemes both demonstrated excellent validity. The TLISS and TLICS scales both exhibited substantial reliability and validity. However, the TLISS system displayed greater inter-observer correlation than did the TLICS and demonstrated significant improvements in reliability over time.


Surface display of the receptor-binding region of the Lactobacillus brevis S-layer protein in Lactococcus lactis provides nonadhesive lactococci with the ability to adhere to intestinal epithelial cells.
Lactobacillus brevis is a promising lactic acid bacterium for use as a probiotic dietary adjunct and a vaccine vector. The N-terminal region of the S-layer protein (SlpA) of L. brevis ATCC 8287 was recently shown to mediate adhesion to various human cell lines in vitro. In this study, a surface display cassette was constructed on the basis of this SlpA receptor-binding domain, a proteinase spacer, and an autolysin anchor. The cassette was expressed under control of the nisA promoter in Lactococcus lactis NZ9000. Western blot assay of lactococcal cell wall extracts with anti-SlpA antibodies confirmed that the SlpA adhesion domain of the fusion protein was expressed and located within the cell wall layer. Whole-cell enzyme-linked immunosorbent assay and immunofluorescence microscopy verified that the SlpA adhesion-mediating region was accessible on the lactococcal cell surface. In vitro adhesion assays with the human intestinal epithelial cell line Intestine 407 indicated that the recombinant lactococcal cells had gained an ability to adhere to Intestine 407 cells significantly greater than that of wild-type L. lactis NZ9000. Serum inhibition assay further confirmed that adhesion of recombinant lactococci to Intestine 407 cells was indeed mediated by the N terminus-encoding part of the slpA gene. The ability of the receptor-binding region of SlpA to adhere to fibronectin was also confirmed with this lactococcal surface display system. These results show that, with the aid of the receptor-binding region of the L. brevis SlpA protein, the ability to adhere to gut epithelial cells can indeed be transferred to another, nonadhesive, lactic acid bacterium.


Graph Theory on Brain Cortical Sources in Parkinson's Disease: The Analysis of 'Small World' Organization from EEG.
Parkinson's disease (PD) is the second most common neurodegenerative disease in the elderly population. Similarly to other neurodegenerative diseases, the early diagnosis of PD is quite difficult. The current pilot study aimed to explore the differences in brain connectivity between PD and NOrmal eLDerly (Nold) subjects to evaluate whether connectivity analysis may speed up and support early diagnosis. A total of 26 resting state EEGs were analyzed from 13 PD patients and 13 age-matched Nold subjects, applying to cortical reconstructions the graph theory analyses, a mathematical representation of brain architecture. Results showed that PD patients presented a more ordered structure at slow-frequency EEG rhythms (lower value of SW) than Nold subjects, particularly in the theta band, whereas in the high-frequency alpha, PD patients presented more random organization (higher SW) than Nold subjects. The current results suggest that PD could globally modulate the cortical connectivity of the brain, modifying the functional network organization and resulting in motor and non-motor signs. Future studies could validate whether such an approach, based on a low-cost and non-invasive technique, could be useful for early diagnosis, for the follow-up of PD progression, as well as for evaluating pharmacological and neurorehabilitation treatments.


Impact of radiation and hepatitis virus infection on risk of hepatocellular carcinoma.
In cohort studies of atomic bomb survivors and Mayak nuclear facility workers, radiation-associated increases in liver cancer risk were observed, but hepatitis B virus (HBV) and hepatitis C virus (HCV) infections were not taken strictly into account. We identified 359 hepatocellular carcinoma (HCC) cases between 1970 and 2002 in the cohort of atomic bomb survivors and estimated cumulative incidence of HCC by radiation dose. To investigate contributions of radiation exposure and hepatitis virus infection to HCC risk, we conducted a nested case-control study using sera stored before HCC diagnosis in the longitudinal cohort of atomic bomb survivors. The study included 224 HCC cases and 644 controls that were matched to the cases on gender, age, city, and time and method of serum storage, and countermatched on radiation dose. The cumulative incidence of HCC by follow-up time and age increased significantly with radiation dose. The relative risk (RR) of HCC for radiation at 1 Gy was 1.67 (95% confidence interval: 1.22-2.35) with adjustment for alcohol consumption, body mass index (BMI), and smoking habit, whereas the RRs for HBV or HCV infection alone were 63 (20-241) and 83 (36-231) with such adjustment, respectively. Those estimates changed little when radiation and hepatitis virus infection were fit simultaneously. The RR of non-B, non-C HCC at 1 Gy was 1.90 (1.02-3.92) without adjustment for alcohol consumption, BMI, or smoking habit and 2.74 (1.26-7.04) with such adjustment. These results indicate that radiation exposure and HBV and HCV infection are associated independently with increased HCC risk. In particular, radiation exposure was a significant risk factor for non-B, non-C HCC with no apparent confounding by alcohol consumption, BMI, or smoking habit.


The effects of P2 segment of posterior cerebral artery to thalamus blood supply pattern on gait in cerebral small vessel disease: A 7 T MRI based study.
Gait disturbance is a manifestation of cerebral small vessel disease (CSVD). The posterolateral thalamus (PL), whose blood is mainly supplied by the P2 segment of posterior cerebral artery (P2-PCA), plays pivotal roles in gait regulation. We investigated the influence of the distance between P2-PCA and PL on gait with varying CSVD burden. 71 participants were divided into low and high CSVD burden groups. The distance from P2-PCA to PL was measured using 7 T TOF-MRA and categorized into an immediate or distant PCA-to-thalamus pattern. Functional connectivity (FC) and voxel-based morphometry were assessed to evaluate functional and structural alterations. In the low CSVD burden group, immediate PCA-to-thalamus supply strongly correlates with longer step length and higher wave phase time percent, and exhibited enhanced FCs in left supplementary motor area, right precentral cortex (PreCG.R). While in the high CSVD burden group, no association between PCA-to-thalamus pattern and gait was found, and we observed reduced FC in PreCG.R with immediate PCA-to-thalamus pattern. Higher CSVD burden was associated with decreased gray matter density in bilateral thalamus. However, no significant structural thalamic change was observed between the two types of PCA-to-thalamus patterns in all patients. Our study demonstrated patients with immediate PCA-to-thalamus supply exhibited better gait performance in low CSVD burden populations, which also correlated with enhanced FCs in motor-related cortex, indicating the beneficial effects of the immediate PCA-to-thalamus supply pattern. In the higher burden CSVD populations, the effects of PCA-to-thalamus pattern on gait are void, attributable to the CSVD-related thalamic destruction and impairment of thalamus-related FC.


Cirrhosis associated with pyridoxal 5'-phosphate treatment of pyridoxamine 5'-phosphate oxidase deficiency.
We report the case of an 8-year-old boy with pyridoxamine 5'-phosphate oxidase (PNPO) deficiency. He developed seizures at 24 h of age that were refractory to standard anticonvulsant therapy and a trial of pyridoxine but responded to pyridoxal phosphate (PLP) at 28 days of life. Genetic testing identified compound heterozygous mutations in the PNPO gene. Management of encephalopathic episodes required escalation of PLP dose to 100 mg/kg/day by 2 years of age. Routine blood tests at this time showed significantly deranged liver function tests (LFTs). A wedge liver biopsy showed early cirrhosis with marked elevation of pyridoxal and pyridoxic acid levels in the liver sample. Despite extensive investigation, no cause other than PLP therapy could be identified for the cirrhosis. The PLP dose was weaned to 50 mg/kg/day before episodes of encephalopathy recurred. Concurrent with the reduction of his PLP dose, LFTs showed improvement. However, at 8 years of age, there is persistent evidence of hepatic fibrosis and early portal hypertension. We hypothesise that hepatic toxicity due to PLP or its degradation products is the cause of cirrhosis in this boy. Until further evidence becomes available, we would suggest that people with PNPO deficiency are treated with the minimum dose of PLP required to prevent episodes of encephalopathy.


Thyroid autoimmunity and recurrent spontaneous abortion in Iran: a case-control study.
To determine the association of thyroglobulin antibodies (TG-Ab) and thyroid peroxidase antibodies (TPO-Ab) with recurrent spontaneous abortion in a euthyroid, nonpregnant population of women in Iran. In this case-control study conducted between November 2003 and September 2006 in Tehran, Iran, nonpregnant women with a history of 3 or more consecutive pregnancy losses and age-matched, healthy parous women without a history of reproductive problems were assessed. Thyroid function tests were performed, which included assessment of thyroid-stimulating hormone, triiodothyronine, thyroxine, and the presence of TG-Ab and TPO-Ab. A total of 641 patients and 269 controls were included. Mean age (+/- SD) was 30.6 +/- 6.4 years (range, 16-51 years) in the patient group and 30.05 +/- 6.6 years (range, 18-48 years) in the control group. Thyroid antibodies were present in 157 of 641 patients (24.5%) and in 34 of 269 controls (12.6%) (P<.001). The presence of thyroid antibodies was significantly associated with recurrent abortion independent of the impact of age with an odds ratio of 2.24 (95% confidence interval, 1.5-3.35). In this population of women in Iran, TG-Ab and TPO-Ab were identified more frequently in women with recurrent abortions compared with controls, and thyroid autoimmunity was independently associated with a higher risk of recurrent abortion.


Biology of human papillomavirus (HPV) infections and their role in squamous cell carcinogenesis.
Current data implicating the role of human papillomavirus (HPV) infections in squamous cell carcinogenesis may be summarised as follows: animal models have shown that PVs can induce malignant transformation; HPV involvement in both benign and malignant human squamous cell tumours has been demonstrated by morphological, immunohistochemical and DNA hybridisation techniques; HPV infections in the genital tract are venereally transmitted and are associated with the same risk factors as cervical carcinoma; the natural history of cervical HPV lesions is similar to that of CIN, namely, they have the potential to develop into carcinoma in situ; malignant transformation of PV-induced lesions seems to depend on virus type and the physical state of its DNA, e.g., whether or not it is integrated in the host cell DNA; malignant transformation most probably requires synergistic actions between the PVs and chemical or physical carcinogens, or other infectious agents; genetic disposition (at least in animals) significantly contributes to malignant transformation; immunological defence mechanisms of the host are probably capable of modifying the course of PV infections (efficacy in man remains to be elucidated). Many details of the molecular mechanisms, however, still remain to be clarified. Although BPV1 is capable of transforming fibroblasts, the way that papillomaviruses transform epithelial cells is unclear. Which gene is capable of inducing the limited cell proliferation in benign lesions? No model systems exist to provide the answer nor to elucidate the progression to malignant cells and then to invasive cancer. Improved tissue culture systems for in vitro differentiation of keratinocytes should help in elucidating the biology of papillomaviruses and their interaction with cell division and differentiation.


Development of machine learning models for the prediction of long-term feeding tube dependence after oral and oropharyngeal cancer surgery.
To predict the necessity of enteral nutrition at 28 days after surgery in patients undergoing major head and neck oncologic procedures for oral and oropharyngeal cancers. Data from 193 patients with oral cavity and oropharyngeal squamous cell carcinoma were retrospectively collected at two tertiary referral centers to train (n = 135) and validate (n = 58) six supervised machine learning (ML) models for binary prediction employing 29 clinical variables available pre-operatively. The accuracy of the six ML models ranged between 0.74 and 0.88, while the measured area under the curve (AUC) between 0.75 and 0.87. The ML algorithms showed high specificity (range 0.87-0.96) and moderate sensitivity (range: 0.31-0.77) in detecting patients with ≥28 days feeding tube dependence. Negative predictive value was higher (range: 0.81-0.93) compared to positive predictive value (range: 0.40-0.71). Finally, the F1 score ranged between 0.35 and 0.74. Classification performance of the ML algorithms showed optimistic accuracy in the prediction of enteral nutrition at 28 days after surgery. Prospective studies are mandatory to define the clinical benefit of a ML-based pre-operative prediction of a personalized nutrition protocol.


Asthma and diving.
Medical opinions conflict as to whether the mild, or infrequent, asthmatic should dive. However, even an asymptomatic asthmatic may have small airway obstruction.


Laparoscopic versus open posterior adrenalectomy: a case-control study of 100 patients.
Few controlled studies have compared laparoscopic transabdominal adrenalectomy (LA) to conventional open posterior adrenalectomy (PA). Five patients have undergone successful LA at our institution between 1992 and 1996. A matched case-control study of 50 PA patients was performed during a similar time period. Follow-up was complete in 82% of patients with a mean follow-up time of 25 months. There were no statistically significant differences between the LA and PA groups with regard to the following demographic features: age, gender, endocrine disorder, side and size of tumor, and body habitus. Statistically significant differences (p < 0.05), however, were present (LA vs PA) when we compared the following results: mean hospital stay (3.1 versus 5.7 days), narcotic equivalents (28 versus 48), return to normal activity (3.8 versus 7 weeks), patient satisfaction (9 versus 7 [scale 1 to 10, 10 being most satisfied]), late morbidity (0 versus 54%), and operating room time (167 versus 127 minutes). Median hospital charges ($7,000 versus $6,000) were slightly higher in the LA group (p = 0.05). Although LA is technically more demanding and slightly more expensive to perform, advances appear to exist for LA with regard to patient comfort, patient satisfaction, hospital stay, and return to normal daily activities. Late incisional complications are dramatically less in the LA group.


Central auditory deficits associated with genetic forms of peripheral deafness.
Since the 1990s, the study of inherited hearing disorders, mostly those detected at birth, in the prelingual period or in young adults, has led to the identification of their causal genes. The genes responsible for more than 140 isolated (non-syndromic) and about 400 syndromic forms of deafness have already been discovered. Studies of mouse models of these monogenic forms of deafness have provided considerable insight into the molecular mechanisms of hearing, particularly those involved in the development and/or physiology of the auditory sensory organ, the cochlea. In parallel, studies of these models have also made it possible to decipher the pathophysiological mechanisms underlying hearing impairment. This has led a number of laboratories to investigate the potential of gene therapy for curing these forms of deafness. Proof-of-concept has now been obtained for the treatment of several forms of deafness in mouse models, paving the way for clinical trials of cochlear gene therapy in patients in the near future. Nevertheless, peripheral deafness may also be associated with central auditory dysfunctions and may extend well beyond the auditory system itself, as a consequence of alterations to the encoded sensory inputs or involvement of the causal deafness genes in the development and/or functioning of central auditory circuits. Investigating the diversity, causes and underlying mechanisms of these central dysfunctions, the ways in which they could impede the expected benefits of hearing restoration by peripheral gene therapy, and determining how these problems could be remedied is becoming a research field in its own right. Here, we provide an overview of the current knowledge about the central deficits associated with genetic forms of deafness.


Efficacy and Cost-Benefit Analysis of a Global Environmental Cleaning Algorithm on Hospital-Acquired Infection Rates.
This study evaluates clinical outcomes and cost-benefit analysis before and after implementation of a global environmental cleaning algorithm on all hospital-acquired infection (HAI) rates. A retrospective, quasi-experimental study design was used to review the hospital's procedure and infection rate database for all HAIs from January 1, 2009, through June 30, 2011. We calculated the infection rates and did a cost-benefit analysis before and after the environmental cleaning algorithm was instituted on July 19, 2010. The methicillin-resistant Staphylococcus aureus rates per 1000 patient days decreased 63%. The central line-associated bloodstream infection rate had a 72% reduction. The catheter-associated urinary tract infection rate dropped 79%. The vancomycin-resistant Enterococcus rate went down 53%. The hospital-acquired Acinetobacter baumanii infection rate had a 65% reduction. The medical intensive care unit ventilator-associated pneumonia rate was reduced 72%. Cardiothoracic sternal wound surgical site infection (SSI) rate dropped 93%, spinal fusion SSI decreased 56%, and total knee arthroplasty SSI was eliminated with a 100% reduction. The hospital avoided an estimated 13 deaths and $5,800,526 in costs during a 1-year period. This global environmental cleaning protocol was associated with decreased HAIs and hospital costs.


Topoisomerase II alpha status in renal medullary carcinoma: immuno-expression and gene copy alterations of a potential target of therapy.
Renal medullary carcinoma is an aggressive renal neoplasm without currently available effective therapy to our knowledge. Topoisomerase II alpha is a gyrase involved in cell proliferation, and DNA maintenance and repair. Topoisomerase II alpha is a target of inhibiting agents such as anthracyclines. Triggered by a recent response to topoisomerase II alpha inhibitors in a patient with renal medullary carcinoma, we evaluated topoisomerase II alpha expression in relation to the proliferation index and topoisomerase II alpha gene copy number status in a larger series of patients with renal medullary carcinoma. Archival tissues from 14 renal medullary carcinomas were retrieved from our 3 institutions. Immunohistochemistry was performed using monoclonal antibodies for topoisomerase II alpha and Ki67. The percent of cells with positive nuclear staining was assessed in the highest area of expression for each marker. A previously suggested greater than 5% cutoff was used for topoisomerase II alpha over expression. The topoisomerase II alpha gene copy number was evaluated using fluorescence in situ hybridization. Locus specific topoisomerase II alpha gene and chromosome 17 centromere probes were used. The total number of topoisomerase II alpha and chromosome 17 centromere signals was counted in 150 cells per tumor and a topoisomerase II alpha-to-chromosome 17 centromere signal ratio was calculated in each tumor. A topoisomerase II alpha-to-chromosome 17 centromere ratio of 2.0 or greater and less than 0.8 was used as a cutoff for amplification and deletion, respectively. The percent of tumor cells with polysomic, eusomic or monosomic chromosome 17 status was also determined. On immuno-expression analysis topoisomerase II alpha immunohistochemistry was technically inconclusive in 1 renal medullary carcinoma. Topoisomerase II alpha was over expressed in 11 of 13 renal medullary carcinomas (85%) (median 50%, range 1% to 80%). As expected, a high Ki67 proliferation index was noted in 13 of 14 tumors (median 87.5%, range 2% to 100%). Ki67 expression was greater than topoisomerase II alpha expression in all 13 informative tumors. A strong, statistically significant correlation was found for topoisomerase II alpha and Ki67 expression (pairwise CC 0.9, p = 0.0000). Topoisomerase II alpha over expression was associated with shorter survival (p = 0.000). On fluorescence in situ hybridization no topoisomerase II alpha amplification was detected in any of the 14 renal medullary carcinomas, including the 11 with topoisomerase II alpha over expression. Topoisomerase II alpha gene deletions were noted in 4 tumors. Two of 4 deletions were associated with chromosome 17 monosomy and 2 were in eusomic chromosome 17 tumors. Topoisomerase II alpha is over expressed in 85% of renal medullary carcinomas, potentially supporting the use of topoisomerase II alpha inhibitor agents to treat this aggressive renal tumor. Our findings suggest that topoisomerase II alpha over expression in our renal medullary carcinoma cohort was not due to gene amplification, but rather to transcriptional or post-transcriptional modifications. The significance of the incidentally found topoisomerase II alpha deletions in 28% of renal medullary carcinomas requires further evaluation.


Endovascular Treatment of Posterior Cerebral Artery Aneurysms With Flow Diversion: Case Series and Systematic Review.
Flow diversion of posterior cerebral artery (PCA) aneurysms has not been widely reported, possibly owing to concerns regarding parent vessel size and branch vessel coverage. To examine the safety and effectiveness of PCA aneurysm flow diverter treatment. Retrospective review of PCA aneurysms treated with the Pipeline Embolization Device (PED; Medtronic Inc, Dublin, Ireland) at 3 neurovascular centers, including periprocedural complications and clinical and angiographic outcomes. Systematic review of the literature identified published reports of PCA aneurysms treated with flow diversion. Rates of aneurysm occlusion and complications were calculated, and outcomes of saccular and fusiform aneurysm treatments were compared. Ten PCA aneurysms in 9 patients were treated with the PED. There were 2 intraprocedural thromboembolic events (20%), including 1 symptomatic infarction and 1 delayed PED thrombosis. Eight of 10 patients returned to or improved from their baseline functional status. Complete aneurysm occlusion with parent vessel preservation was achieved in 75% (6/8) of cases at mean follow-up of 16.7 mo. Eleven of 12 (92%) major branch vessels covered by a PED remained patent. Including the present study, systematic review of 15 studies found a complete aneurysm occlusion rate of 88% (30/34) and complication rate of 26% (10/38), including 5 symptomatic ischemic strokes (13%; 5/38). Fusiform aneurysms more frequently completely occluded compared with saccular aneurysms (100% vs 70%; P = .03) but were associated with a higher complication rate (43% vs 9%; P = .06). The safety and effectiveness profile of flow diverter treatment of PCA aneurysms may be acceptable in select cases.


[Growth of Azotobacter chroococcum strains on different substrates].
Azotobacter chroococcum 34, actively growing on alpha-ketoglutaric acid, and Azotobacter chroococcum B and KL, which almost do not assimilate this acid, can grow on the majority of substrates of the tricarboxylic acid cycle and on glucose, with and without nitrogen. The rate of assimilation of alpha-ketoglutaric acid is several times higher in the cells cultivated in the presence of this acid than in the cells grown on other substrates. This difference seems to involve the mechanism of transport of alpha-ketoglutaric acid into the cell.


Nicotinic acid transport by brush border membrane vesicles from rabbit kidney.
The transport of nicotinic acid was investigated in brush border membrane vesicles isolated from rabbit kidney. The imposition of a Na+ gradient (out to in) induced a transient stimulation of nicotinic acid uptake above its final equilibrium value. This stimulation was specific for Na+. The uptake of nicotinic acid by the brush border membranes represented transport into an internal space and occurred in the absence of significant nicotinic acid degradation. The Na+ gradient-dependent uptake of nicotinic acid was saturable, apparent Km = 0.3 mM. Uptake of nicotinic acid was inhibited by its two isomers: picolinic and isonicotinic acid. In contrast, pyridine derivatives with two carboxyl groups or an amide group in addition to the carboxyl group were without inhibitory effect. Evaluation of changes in membrane potential using the lipophilic cation triphenylmethylphosphonium demonstrated that conditions that transiently generated either an interior-positive or an interior-negative membrane potential failed to affect the Na+-dependent transport of nicotinic acid. These findings provide evidence of the existence on the luminal membrane of a Na+ gradient-dependent and electroneutral transport system for nicotinic acid.


Cytochrome p450sky interacts directly with the nonribosomal peptide synthetase to generate three amino acid precursors in skyllamycin biosynthesis.
The generation of modified amino acid precursors for incorporation in nonribosomal peptide synthesis (NRPS) plays a crucial, if often understated, role in the generation of peptide natural products. The biosynthesis of the cyclic depsipeptide skyllamycin requires three β-hydroxylated amino acid precursors, with in vivo gene inactivation experiments implicating cytochrome P450sky (CYP163B3) in the hydroxylation of these amino acids. Here, we demonstrate the in vitro oxidation of l-amino acid substrates bound to peptidyl carrier protein (PCP) domains 5, 7, and 11 of the skyllamycin nonribosomal synthetase by P450sky. Selectivity for these domains over other PCP domains could be demonstrated, with hydroxylation selective for l-amino acids and stereospecific in nature resulting in the (2S,3S)-configuration. The oxidation of amino acids or small molecule substrate analogues was not supported, demonstrating the necessity of the carrier protein in P450sky-catalyzed hydroxylation. The binding of aminoacyl-PCP substrates to P450sky was detected for the catalytically active PCP7 but not for the catalytically inactive PCP10, indicating carrier protein-mediated selectivity in P450sky substrate binding. X-ray crystal structures of P450sky reveal a 3D-structure with a highly open active site, the size of which is dictated by the carrier protein bound nature of the substrate. P450sky is the first P450 demonstrated to not only interact directly with PCP-bound amino acids within the peptide-forming NRPS but also to do so with three different PCP domains in a specific fashion. This represents an expansion of the complexity and scope of NRPS-mediated peptide synthesis, with the generation of hydroxylated amino acid precursors occurring through the interaction of P450 enzymes following, rather than prior to, the selection of amino acids by NRPS-adenylation domains.


Subtle changes in daily functioning predict conversion from normal to mild cognitive impairment or dementia: an analysis of the NACC database.
There are relatively small but observable changes in functional ability in those without Mild cognitive impairment (MCI) or dementia. The present study seeks to understand whether these individuals go on to develop MCI or dementia by assessing the association between baseline Functional Activities Questionnaire (FAQ) and conversion independent and after adjustment for cognitive tests. The NACC database was used to conduct the analysis of which 7,625 participants were initially identified as having more than one visit and who were cognitively normal at their first visit. Cox proportional hazards were used to fit three models that controlled for executive and non-executive cognitive domains. A similar model was used to assess the effect of FAQ subcategories on conversion. Of these individuals, 1,328 converted to either MCI or dementia by visit 10. Converters had a total visit 1 FAQ score significantly higher than non-converters indicating more functional impairment at baseline. After adjustment for cognitive tests, the association between visit 1 FAQ and subsequent conversion was not attenuated. Doing taxes, remembering dates, and traveling were individually identified as significant predictors of conversion. The FAQ can be used as an indirect measure of functional ability and is associated with conversion to MCI or dementia. There is a selective and significant association between changes in financial ability and conversion that is in accordance with other research of financial capacity.


[Molecular characterization of beta-lactamase-associated resistance in Acinetobacter baumannii strains isolated from clinical samples].
Acinetobacter baumannii is an important cause of nosocomial infections that particularly increase the mortality and the morbidity at the intensive care units of the hospitals. The aims of this study were to evaluate the resistance genes, antibiotic susceptibility and the clonal relations among Acinetobacter strains isolated from clinical samples and to determine the resistance mechanisms related to these bacteria in our hospital. A total of 201 A.baumannii strains isolated from different clinical samples (35.3% from tracheal aspirate, 27.3% from blood, 18.4% from abscess material, 19% from other samples) of 160 inpatients evaluated at the Ibni Sina Hospital Central Bacteriology Laboratory, Ankara University School of Medicine, Turkey from April 2010 to December 2011, were included in the study. Identification of the isolates and their susceptibility testing against amikacin, ciprofloxacin, tetracycline, sulbactam/ampicillin, trimethoprim/sulfametoxazole (SXT), ceftazidime, gentamicin, imipenem, levofloxacin, meropenem, piperacillin/tazobactam, cefoperazone/sulbactam, cefepime and colistin were performed by the automated systems, namely Vitek 2 (bioMérieux, France) and BD Phoenix (Becton Dickinson, USA). The molecular mechanisms of beta-lactamase resistance and the presence of integrons were analyzed by polymerase chain reaction (PCR). Moreover, since blaPER-1 gene is of high frequency in Turkey, it was also investigated in the isolates. Pulsed-field gel electrophoresis (PFGE) was performed to examine the clonal relations between isolates. Our results indicated that multidrug resistance rate of A.baumannii was 94.5% (190/201), while 94% (189/201) of the isolates were susceptible to colistin thus making it the most potent antimicrobial agent, followed by amikacin and SXT with a susceptibility rate of 32%. Twelve colistin-resistant isolates were further investigated with the E-test method (AB Biodisk, Sweden) and found to be colistin-resistant. While the results were negative for the genes responsible from metallo-beta-lactamase production, positive results were obtained for blaOXA genes at various rates (OXA-51 100%; OXA-23 91.5%; OXA-58 7%; OXA-24 2%). PFGE results revealed four different main clones (29 isolates in genotype A, 23 in genotype B, 18 in genotype C and 7 in genotype D) in the study population. No common epidemic isolate was detected. Class 1 integrons which take part in the transfer of resistance genes were detected in 112 (55.7%) isolates. There was no statistically significant difference between the genotype distributions of class 1 integron positive strains (p> 0.05). The relationship between the presence of integron in multidrug resistant isolates and resistance to tetracyclin, SXT, imipenem, meropenem, cefoperazone/sulbactam and cefepime were found to be statistically significant (p< 0.05). Of the isolates 42 (21%) were positive for blaPER- 1 gene and all were resistant to ceftazidime. This study indicated that blaOXA genes found together with blaOXA-51 genes play an important role in carbapenem resistance of A.baumannii strains. Moreover, multidrug resistance is still an important problem in infections caused by A.baumannii and integrons play a role in the transfer of the resistance genes. In conclusion, multidrug resistant A.baumannii strains were common in our hospital and our epidemiologic data would be helpful for further investigations and in therapeutical approaches.


Viewing cancer genes from co-evolving gene modules.
Studying the evolutionary conservation of cancer genes can improve our understanding of the genetic basis of human cancers. Functionally related proteins encoded by genes tend to interact with each other in a modular fashion, which may affect both the mode and tempo of their evolution. In the human PPI network, we searched for subnetworks within each of which all proteins have evolved at similar rates since the human and mouse split. Identified at a given co-evolving level, the subnetworks with non-randomly large sizes were defined as co-evolving modules. We showed that proteins within modules tend to be conserved, evolutionarily old and enriched with housekeeping genes, while proteins outside modules tend to be less-conserved, evolutionarily younger and enriched with genes expressed in specific tissues. Viewing cancer genes from co-evolving modules showed that the overall conservation of cancer genes should be mainly attributed to the cancer proteins enriched in the conserved modules. Functional analysis further suggested that cancer proteins within and outside modules might play different roles in carcinogenesis, providing a new hint for studying the mechanism of cancer.


Chronic Activation of Gp1 mGluRs Leads to Distinct Refinement of Neural Network Activity through Non-Canonical p53 and Akt Signaling.
Group 1 metabotropic glutamate receptors (Gp1 mGluRs), including mGluR1 and mGluR5, are critical regulators for neuronal and synaptic plasticity. Dysregulated Gp1 mGluR signaling is observed with various neurologic disorders, including Alzheimer's disease, Parkinson's disease, epilepsy, and autism spectrum disorders (ASDs). It is well established that acute activation of Gp1 mGluRs leads to elevation of neuronal intrinsic excitability and long-term synaptic depression. However, it remains unknown how chronic activation of Gp1 mGluRs can affect neural activity and what molecular mechanisms might be involved. In the current study, we employed a multielectrode array (MEA) recording system to evaluate neural network activity of primary mouse cortical neuron cultures. We demonstrated that chronic activation of Gp1 mGluRs leads to elevation of spontaneous spike frequency while burst activity and cross-electrode synchronization are maintained at the baseline. We further showed that these neural network properties are achieved through proteasomal degradation of Akt that is dependent on the tumor suppressor p53. Genetically knocking down p53 disrupts the elevation of spontaneous spike frequency and alters the burst activity and cross-electrode synchronization following chronic activation of Gp1 mGluRs. Importantly, these deficits can be restored by pharmacologically inhibiting Akt to mimic inactivation of Akt mediated by p53. Together, our findings reveal the effects of chronic activation of Gp1 mGluRs on neural network activity and identify a unique signaling pathway involving p53 and Akt for these effects. Our data can provide insights into constitutively active Gp1 mGluR signaling observed in many neurologic and psychiatric disorders.


Modeling Human Cytomegalovirus in Humanized Mice for Vaccine Testing.
Human cytomegalovirus (HCMV or HHV-5) is a globally spread pathogen with strictly human tropism that establishes a lifelong persistence. After primary infection, high levels of long-term T and B cell responses are elicited, but the virus is not cleared. HCMV persists mainly in hematopoietic reservoirs, whereby occasional viral reactivation and spread are well controlled in immunocompetent hosts. However, when the immune system cannot control viral infections or reactivations, such as with newborns, patients with immune deficiencies, or immune-compromised patients after transplantations, the lytic outbursts can be severely debilitating or lethal. The development of vaccines for immunization of immune-compromised hosts has been challenging. Several vaccine candidates did not reach the potency expected in clinical trials and were not approved. Before anti-HCMV vaccines can be tested pre-clinically in immune-compromised hosts, reliable in vivo models recapitulating HCMV infection might accelerate their clinical translation. Therefore, immune-deficient mouse strains implanted with human cells and tissues and developing a human immune system (HIS) are being explored to test anti-HCMV vaccines. HIS-mice resemble immune-compromised hosts as they are equipped with antiviral human T and B cells, but the immune reactivity is overall low. Several groups have independently shown that HCMV infections and reactivations can be mirrored in HIS mice. However, these models and the analyses employed varied widely. The path forward is to improve human immune reconstitution and standardize the analyses of adaptive responses so that HIS models can be forthrightly used for testing novel generations of anti-HCMV vaccines in the preclinical pipeline.


Overview: animal models of osteopenia and osteoporosis.
Prior to initiating a clinical trial in a post-menopausal osteoporosis study, it is reasonable to recommence the evaluation of treatment in the 9-month-old ovariectomized female rat. A female rat of this age has reached peak bone mass and can be manipulated to simulate clinical findings of post-menopausal osteoporosis. Ample time exists for experimental protocols that either prevent estrogen depletion osteopenia or restore bone loss after estrogen depletion. More time can be saved by acceleration of the development of the osteopenia by combining ovariectomized (OVX) plus immobilization (IM) models. Methods like serum biochemistry, histomorphometry and densitometry used in humans are applicable in rats. Like most animal models of osteopenia, the rat develops no fragility fractures, but mechanical testing of rat bones substitutes as a predictor of bone fragility. Recent studies have shown that the prevailing activity in cancellous and cortical bone of the sampling sites in rats is remodeling. The problems of dealing with a growing skeleton, the site specificity of the OVX and IM models, the lack of trabecular and Haversian remodeling and the slow developing cortical bone loss have been and can be overcome by adding beginning and pre-treatment controls and muscle mass measurements in all experimental designs, selecting cancellous bone sampling sites that are remodeling, concentrating the analysis of cortical bone loss to the peri-medullary bone and combining OVX and IM in a model to accelerate the development of both cancellous and cortical bone osteopenia. Not to be forgotten is the distal tibia site, an adult bone site with growth plate closure at 3 months and low trabecular bone turnover and architecture similar to human spongiosa. This site would be most challenging to the action of bone anabolic agents. Data about estrogen-deplete mice are encouraging, but the ovariectomized rat model suggests that developing an ovariectomized mouse model as an alternative is not urgent. Nevertheless, the mouse model has a place in drug development and skeletal research. In dealing with drug development, it could be a useful model because it is a much smaller animal requiring fewer drugs for screening. In skeletal research mice are useful in revealing genetic markers for peak bone mass and gene manipulations that affect bone mass, structure and strength. When the exciting mouse glucocorticoid-induced bone loss model of Weinstein and Manolagas is confirmed by others, it could be a significant breakthrough for that area of research. Lastly, we find that the information generated from skeletal studies of nonhuman primates has been most disappointing and recommend that these expensive skeletal studies be curtailed unless it is required by a regulatory agency for safety studies.


Using High Sensitivity Troponins to Rule Out Acute Coronary Syndrome and Lower Admission Rates.
Chest pain is a common complaint encountered by Emergency Medicine physicians in the emergency department (ED). History taking, electrocardiograms, and cardiac biomarkers are the mainstays of the evaluation process of patients who present to the ED with symptoms of an acute coronary syndrome. Cardiac troponin is the primary biomarker used for the diagnosis of acute myocardial infarction. In January 2017, high sensitivity cardiac troponins (hs-cTns) were approved for use in the United States. These markers have increased sensitivity and can more rapidly detect myocardial injury, making them very useful in the ED. However, despite improved sensitivity, elevations in hs-cTn can occur in a large number of patients who are not currently experiencing an acute myocardial infarction. As a result, it is important that clinicians understand the value of serial cardiac biomarker measurements and how to incorporate hs-cTn levels below the limit of detection into clinical decision-making. A large percentage of these low-risk patients end up having negative cardiac work-ups. This article will include a discussion on how to utilize common risk stratification tools in the ED to identify cohorts of patients suitable for discharge without additional testing. ED physicians must understand the limitations and benefits of hs-cTn levels and how to incorporate the information obtained from these biomarkers into risk scores in order to strengthen disposition decisions and safely discharge patients from the ED.


[Therapeutic efficacy of hematopoietic stem cell transplantation in patients with chronic myelogenous leukemia].
To evaluate the clineffects of autologous and allogeneic hematopoietic stem cell transplantation (HSCT) for chronic myelogenous leukemia (CML). Fifty-seven patients with CML were treated by HSCT, including 8 patients treated with autologous transplantation in vivo and vitro purging minimal residual disease (MRD), 39 with related donor allogeneic HSCT (allo-HSCT), and 10 with unrelated donor allo-HSCT. For the conditioning regimen, total-body irradiation with cyclophosphamide (CTX) was given in 32 patients, modified BuCY protocol (hydroxyurea, busulfan, Ara-C, CTX) in 24 patients, and MACC protocol (melphalan, Ara-C, CTX and lomustine) in one patient. Cyclosporine (CsA) and methotrexate (MTX) were used in patients with related donor allo-HSCT, and the combination of CsA, MTX , mycophenolate mofetil (MMF), and antithymocyte globulin (ATG) in unrelated donor all-HSCT to prevent graft versus host disease (GVHD). Kaplan-Meier survival analysis model was used to estimate the overall survival and the disease-free survival (DFS) at 5 years after transplantation. In 8 patients with autologous transplantation, 7 obtained partial or complete cytogenetic remission (CR) within 3 months after transplantation and one died of transplantation-related complication. In 49 patients with allo-HSCT transplantation, all patients obtained CR except for two patients, one of whom failed to obtain CR and the other died of hepatic veno-occlusive disease. The incidence of infection and veno-occlusive disease during transplantation was 33.3% and 7.0%, respectively. The incidence of hemorrhagic cystitis and cytomegalovirus interstitial pneumonia after transplantation was 22.8% and 8.8%, respectively. Veno-occlusive disease, hemorrhagic cystitis or cytomegalovirus interstitial pneumonia did not occur in patients with autologous transplantation. The incidence of acute GVHD was 41.0% and 48.6%, and that of chronic GVHD 40.0% and 42.9% in patients with related and unrelated transplantation, respectively. The rate of relapse was 57.1% and 12.8%, with DFS at 5 years of 25.0% and 61.7%, respectively, in patients with autologous and related donor transplantation. The DFS at 5 years was 70.7% and 34.1%, respectively, in patients with chronic/accelerated phases and blast crisis be fore trans plantation. allo-HSCT can produce a higher clinical cure rate in CML patients in chronic phase CsA+MTX+MMF+ ATG protocol is more effective for prevention and alleviation of acute GVHD in patients with unrelated donor transplantation. Autologous transplantation with bone marrow purging helps prolong the patients' survival and even obtain clinical cure of CML.


P transposons controlled by the heat shock promoter.
We have transformed Drosophila melanogaster with modified P-element transposons, which express the transposase function from the heat-inducible hsp70 heat shock promoter. The Icarus transposon, which contains a direct hsp70-P fusion gene, behaved like a very active autonomous P element even before heat shock induction. Although heat shock led to abundant somatic transcription, transposition of the Icarus element was confined to germ line cells. To reduce the constitutive transposase activity observed for the Icarus element, we attenuated the translational efficiency of the transposase RNA by inserting the transposon 5 neomycin resistance gene between the hsp70 promoter and the P-element sequences. The resulting construct, called Icarus-neo, conferred resistance to G418, and its transposition was significantly stimulated by heat shock. Heat shocks applied during the embryonic or third instar larval stage had similar effects, indicating that transposition of P elements is not restricted to a certain developmental stage. Both Icarus and Icarus-neo destabilized snw in a P-cytotype background and thus at least partially overcome the repression of transposition. Our results suggest that the regulation of P-element transposition occurs at both the transcriptional and posttranscriptional levels.


The rs8099917 polymorphism, when determined by a suitable genotyping method, is a better predictor for response to pegylated alpha interferon/ribavirin therapy in Japanese patients than other single nucleotide polymorphisms associated with interleukin-28B.
We focused on determining the most accurate and convenient genotyping methods and most appropriate single nucleotide polymorphism (SNP) among four such polymorphisms associated with interleukin-28B (IL-28B) in order to design tailor-made therapy for patients with chronic hepatitis C virus (HCV) patients. First, five different methods (direct sequencing, high-resolution melting analysis [HRM], hybridization probe [HP], the InvaderPlus assay [Invader], and the TaqMan SNP genotyping assay [TaqMan]) were developed for genotyping four SNPs (rs11881222, rs8103142, rs8099917, and rs12979860) associated with IL-28B, and their accuracies were compared for 292 Japanese patients. Next, the four SNPs associated with IL-28B were genotyped by Invader for 416 additional Japanese patients, and the response to pegylated interferon/ribavirin (PEG-IFN/RBV) treatment was evaluated when the four SNPs were not in linkage disequilibrium (LD). HRM failed to genotype one of the four SNPs in five patients. In 2 of 287 patients, the results of genotyping rs8099917 by direct sequencing differed from the results of the other three methods. The HP, TaqMan, and Invader methods were accurate for determination of the SNPs associated with IL-28B. In 10 of the 708 (1.4%) patients, the four SNPs were not in LD. Eight of nine (88.9%) patients whose rs8099917 was homozygous for the major allele were virological responders, even though one or more of the other SNPs were heterozygous. The HP, TaqMan, and Invader methods were suitable to determine the SNPs associated with IL-28B. The rs8099917 polymorphism should be the best predictor for the response to the PEG-IFN/RBV treatment among Japanese chronic hepatitis C patients.


Whole mouse brain structural connectomics using magnetic resonance histology.
Diffusion tensor histology holds great promise for quantitative characterization of structural connectivity in mouse models of neurological and psychiatric conditions. There has been extensive study in both the clinical and preclinical domains on the complex tradeoffs between the spatial resolution, the number of samples in diffusion q-space, scan time, and the reliability of the resultant data. We describe here a method for accelerating the acquisition of diffusion MRI data to support quantitative connectivity measurements in the whole mouse brain using compressed sensing (CS). The use of CS allows substantial increase in spatial resolution and/or reduction in scan time. Compared to the fully sampled results at the same scan time, the subtle anatomical details of the brain, such as cortical layers, dentate gyrus, and cerebellum, were better visualized using CS due to the higher spatial resolution. Compared to the fully sampled results at the same spatial resolution, the scalar diffusion metrics, including fractional anisotropy (FA) and mean diffusivity (MD), showed consistently low error across the whole brain (< 6.0%) even with 8.0 times acceleration. The node properties of connectivity (strength, cluster coefficient, eigenvector centrality, and local efficiency) demonstrated correlation of better than 95.0% between accelerated and fully sampled connectomes. The acceleration will enable routine application of this technology to a wide range of mouse models of neurologic diseases.


Unresolved grief in the family.
Unresolved grief after a death in the family may have serious consequences not merely for individuals but for entire families, from grandparents to grandchildren. When the grieving process is abnormal or incomplete, the problem may manifest itself in the physician's office. Observation of a family system can reveal the extent of pathology and the nature of the blocks that keep grief alive. Treatment of unresolved grief ranges from a simple statement of the problem to extended therapy. The physician must be aware of this problem in order to manage it; the cases in this article illustrate presentations of unresolved grief in a variety of family members.


[Immunoglobulins and dysglobulinemias. Introduction and general data].
The author reviews the present state of knowledge of value in rheumatology concerning serum immunoglobulins, their structure, their cellular biosynthesis, and their relation to membrane immunoglobulins of the B lymphocytes. He then examines the concept of dysglobulinaemia and explains why the terms monoclonal immunoglobulin and monoclonal immunoglobulinopathy are preferable to dysglobulinaemia and paraprotein. The clinical and immunochemical classification of monoclonal immunoglobulins, the problem of amylosis, and the relationship with plasmocytic dyscrasias and with the cryoglobulins are then discussed. Afterwards the pathogeny of these affections is briefly considered.


Colony formation of clone-sorted human hematopoietic progenitors.
To characterize human hematopoietic progenitors, we performed methylcellulose cultures of single cells isolated from a population of CD34+ cells by fluorescence-activated cell-sorting (FACS) clone-sorting system. CD34+ cells were detected in bone marrow (BM) and peripheral blood (PB) cells at incidences of 1.0% and 0.01% of total mononuclear cells, respectively. Single cell cultures revealed that approximately 37% of BM CD34+ cells formed colonies in the presence of phytohemagglutinin-leukocyte conditioned medium and erythropoietin. Erythroid bursts-, granulocyte-macrophage (GM) colony-, and pure macrophage (Mac) colony-forming cells were 10% each in CD34+ cells. Approximately 15% of PB CD34+ cells formed colonies in which erythroid bursts were predominant. CD34+ cells were heterogeneous and fractionated by several antibodies in FACS multicolor analysis. In these fractionated cells, CD34+, CD33+ cells formed GM and Mac colonies 7 to 10 times as often as CD34+, CD33- cells. Most of the erythroid bursts and colonies were observed in the fraction of CD34+, CD13- cells or CD34+, CD33- cells. The expression of HLA-DR on CD34+ cells was not related to the incidence, size, or type of colonies. There was no difference in the phenotypical heterogeneity of CD34+ cells between BM and PB. About 10% of CD34+ cells were able to form G colonies in response to granulocyte colony-stimulating factor (G-CSF) and to form Mac colonies in GM-CSF or interleukin-3 (IL-3). Progenitors capable of generating colonies by stimulation of G-CSF were more enriched in CD34+, CD33+ fraction than in CD34+, CD33- fraction. Thus, single cell cultures using the FACS clone-sorting system provide an accurate estimation of hematopoietic progenitors and an assay system for direct action of colony-stimulating factors.


Structural basis for the requirement of additional factors for MLL1 SET domain activity and recognition of epigenetic marks.
The mixed-lineage leukemia protein MLL1 is a transcriptional regulator with an essential role in early development and hematopoiesis. The biological function of MLL1 is mediated by the histone H3K4 methyltransferase activity of the carboxyl-terminal SET domain. We have determined the crystal structure of the MLL1 SET domain in complex with cofactor product AdoHcy and a histone H3 peptide. This structure indicates that, in order to form a well-ordered active site, a highly variable but essential component of the SET domain must be repositioned. To test this idea, we compared the effect of the addition of MLL complex members on methyltransferase activity and show that both RbBP5 and Ash2L but not Wdr5 stimulate activity. Additionally, we have determined the effect of posttranslational modifications on histone H3 residues downstream and upstream from the target lysine and provide a structural explanation for why H3T3 phosphorylation and H3K9 acetylation regulate activity.


Exploring the impact of oral bacteria remnants on stem cells from the Apical papilla: mineralization potential and inflammatory response.
Bacterial persistence is considered one of the main causal factors for regenerative endodontic treatment (RET) failure in immature permanent teeth. This interference is claimed to be caused by the interaction of bacteria that reside in the root canal with the stem cells that are one of the essentials for RET. The aim of the study was to investigate whether prolonged exposure of stem cells from the apical papilla (SCAP) to bacterial remnants of Fusobacterium nucleatum, Actinomyces gerensceriae, Slackia exigua, Enterococcus faecalis, Peptostreptococcaceae yurii, commonly found in infected traumatized root canals, and the probiotic bacteria Lactobacillus gasseri and Limosilactobacillus reuteri, can alter SCAP's inflammatory response and mineralization potential. To assess the effect of bacterial remnants on SCAP, we used UV-C-inactivated bacteria (as cell wall-associated virulence factors) and bacterial DNA. Histochemical staining using Osteoimage Mineralization Assay and Alizarin Red analysis was performed to study SCAP mineralization, while inflammatory and osteo/odontogenic-related responses of SCAPs were assessed with Multiplex ELISA. We showed that mineralization promotion was greater with UV C-inactivated bacteria compared to bacterial DNA. Immunofluorescence analysis detected that the early mineralization marker alkaline phosphatase (ALP) was increased by the level of E. coli lipopolysaccharide (LPS) positive control in the case of UV-C-inactivated bacteria; meanwhile, DNA treatment decreased the level of ALP compared to the positive control. SCAP's secretome assessed with Multiplex ELISA showed the upregulation of pro-inflammatory factors IL-6, IL-8, GM-CSF, IL-1b, neurotrophic factor BDNF, and angiogenic factor VEGF, induced by UV-C-killed bacteria. The results suggest that long term stimulation (for 21 days) of SCAP with UV-C-inactivated bacteria stimulate their mineralization and inflammatory response, while DNA influence has no such effect, which opens up new ideas about the nature of RET failure.


Is the Time of Atrial Fibrillation Recurrence After Duty-Cycled Radiofrequency Ablation Affected by the Pattern of Pulmonary Vein Reconnections?
Pulmonary vein isolation (PVI) with multielectrode duty-cycled radiofrequency (PVAC) has been shown to be effective in the treatment of atrial fibrillation (AF). We describe pulmonary vein (PV) reconnection at repeat ablation in patients with AF recurrence after PVAC PVI and analyze the correlation between the time of AF recurrence and the observed PV reconnection patterns. Eighty-five patients undergoing a redo PVI for recurrent AF 9.2 ± 3.8 months after an initial PVAC PVI procedure was retrospectively enrolled. A total of 93% had PV reconnections with a mean of 2.97 ± 1.2 reconnected PVs/patients and 75% of formerly isolated PVs were found reconnected. The highest reconnection rates (94%) were observed for left common trunks (CTs). A total of 33% patients had three and four reconnected PVs, respectively, while 7% were without PV reconnection. There was a moderate but significant negative correlation between the time of AF recurrences and the extending of PV reconnections at redo PVI for patients with proven PV reconnection (r = -0.32, P = 0.005), whereas five out of six patients without PV reconnection had recurrences within the first 9 months after PVI. At redo ablation most patients with recurrence of AF after PVAC PVI had PV reconnection(s). Patients with PV reconnection(s) showed a moderate negative correlation between the number of reconnected PVs and the time of AF recurrence with more extensive PV reconnections resulting in earlier PV recurrences after the blanking period. Patients without PV reconnection experienced early AF recurrences, indicating non-PV triggers contribute to AF recurrences in these patients.


[The histological identification of Lugaro cells in the cat cerebellar cortex].
Apart from main classic neuron types there is a small group of neurons known in literature as intermediate Lugaro cells. Two Lugaro cells types are distinguished: fusiform and those with triangular cell body. Fusiform cells vary not only in vertical, horizontal or diagonal position of cell body, but also have certain morphological peculiarities. Axons of some fusiform cells project either to molecular layer or reach Purkinje cells layer or remain at the level of granular layer. Along with it, there are true intermediate cells, described by Lugaro. These cell processes project from layer to layer. Neurons with triangular cell body located at different granular layer levels relate to the second group of Lugaro cells. Due to great extension of their processes Lugaro cells possess projection area ranging from molecular layer to white substance. Lugaro cells are independent neurons, always present in cerebellar cortex.


Prevalence and Occlusal Risk Factors for Fractured Incisors among 11-12-Year-Old Children in the Trinidad and Tobago Population.
This cross-sectional survey was carried out to ascertain the prevalence of fractured incisors in 11-12-year-olds. In addition we explored the relationship with overjet, incompetent lips, incisor inclination and to determine if there was any association with ethnicity and gender. All permanent incisors were examined in 672 children comprising 356 females (53.1%) and 315 males (46.9%). The sample comprised 11-12-year-olds in high schools across Trinidad and Tobago. Statistical analysis was undertaken using Chi-square test, independent t-test, and binary logistic regression. The prevalence of fractured incisors was 18.9%. Boys presented with an increased incidence of fractured incisors than girls. 86.3% of dental trauma was untreated. The average overjet of subjects with fractured incisors was 4.2 mm. 18.62% of subjects with fractured incisors had incompetent lips. The most common malocclusion (18.81%) with fractured incisors was class 2 division 1. The Afro-Trinidadian ethnicity had the highest prevalence of fractured incisors (11.0%) when compared to mixed ethnicities, which was statistically significant. Maxillary central incisors were the most commonly injured teeth. Most patients delayed in seeking dental treatment for fractured incisors in our population. Early orthodontic treatment is recommended to help reduce the risk of dental trauma.


Educational attainment among adult survivors of childhood cancer in Great Britain: a population-based cohort study.
Previous studies of educational attainment among childhood cancer survivors were small, had contradictory findings, and were not population based. This study investigated educational attainment in a large population-based cohort of survivors of all types of childhood cancer in Great Britain. Four levels of educational attainment among 10,183 cancer survivors--degree, teaching qualification, advanced (A') levels, and ordinary (O') levels--were compared with expected levels in the general population. A questionnaire was used to obtain educational attainment data for survivors, and comparable information for the general population was available from the General Household Survey. Factors associated with level of educational attainment achieved by cancer survivors were identified using multivariable logistic regression together with likelihood ratio tests. Logistic regression adjusting for age and sex was used for comparisons with the general population. All statistical tests were two-sided. Childhood cancer survivors had lower educational attainment than the general population (degree: odds ratio [OR] = 0.77, 99% confidence interval [CI] = 0.68 to 0.87; teaching qualification: OR = 0.85, 99% CI = 0.77 to 0.94; A'level: OR = 0.85, 99% CI = 0.78 to 0.93; O'level: OR = 0.81, 99% CI = 0.74 to 0.90; P < .001, all levels). Statistically significant deficits were restricted to central nervous system (CNS) neoplasm and leukemia survivors. For leukemia, only those treated with radiotherapy were considered. Odds ratios for achievement by irradiated CNS tumor survivors were 50%-74% of those for cranially irradiated leukemia or nonirradiated CNS tumor survivors. Survivors at greater risk of poorer educational outcomes included those treated with cranial irradiation, diagnosed with a CNS tumor, older at questionnaire completion, younger at diagnosis, diagnosed with epilepsy, and who were female. Specific groups of childhood cancer survivors achieve lower-than-expected educational attainment. Detailed educational support and implementation of regular cognitive assessment may be indicated for some groups to maximize long-term function.


Three Planctomycetes isolated from biotic surfaces in the Mediterranean Sea and the Pacific Ocean constitute the novel species Symmachiella dynata gen. nov., sp. nov. and Symmachiella macrocystis sp. nov.
Planctomycetes is a phylum of environmentally important bacteria, which also receive significant attention due to their fascinating cell biology. Access to axenic Planctomycete cultures is crucial to study cell biological features within this phylum in further detail. In this study, we characterise three novel strains, Mal52T, Pan258 and CA54T, which were isolated close to the coasts of the islands Mallorca (Spain) and Panarea (Italy), and from Monterey Bay, CA, USA. The three isolates show optimal growth at temperatures between 22 and 24 °C and at pH 7.5, divide by polar budding, lack pigmentation and form strong aggregates in liquid culture. Analysis of five phylogenetic markers suggests that the strains constitute two novel species within a novel genus in the family Planctomycetaceae. The strains Mal52T (DSM 101177T = VKM B-3432T) and Pan258 were assigned to the species Symmachiella dynata gen nov., sp. nov., while strain CA54T (DSM 104301T = VKM B-3450T) forms a separate species of the same genus, for which we propose the name Symmachiella macrocystis sp. nov.


Eszopiclone and Zolpidem Produce Opposite Effects on Hippocampal Ripple Density.
Clinical populations have memory deficits linked to sleep oscillations that can potentially be treated with sleep medications. Eszopiclone and zolpidem (two non-benzodiazepine hypnotics) both enhance sleep spindles. Zolpidem improved sleep-dependent memory consolidation in humans, but eszopiclone did not. These divergent results may reflect that the two drugs have different effects on hippocampal ripple oscillations, which correspond to the reactivation of neuronal ensembles that represent previous waking activity and contribute to memory consolidation. We used extracellular recordings in the CA1 region of rats and systemic dosing of eszopiclone and zolpidem to test the hypothesis that these two drugs differentially affect hippocampal ripples and spike activity. We report evidence that eszopiclone makes ripples sparser, while zolpidem increases ripple density. In addition, eszopiclone led to a drastic decrease in spike firing, both in putative pyramidal cells and interneurons, while zolpidem did not substantially alter spiking. These results provide an explanation of the different effects of eszopiclone and zolpidem on memory in human studies and suggest that sleep medications can be used to regulate hippocampal ripple oscillations, which are causally linked to sleep-dependent memory consolidation.


COVID Protection Behaviors, Mental Health, Risk Perceptions, and Control Beliefs: A Dynamic Temporal Network Analysis of Daily Diary Data.
To control infections, behavioral non-pharmaceutical interventions (NPIs) such as social distancing and hygiene measures (masking, hand hygiene) were implemented widely during the COVID-19 pandemic. At the same time, adherence to NPIs has also been implied in an increase in mental health problems. However, the designs of many existing studies are often poorly suited to disentangle complex relationships between NPI adherence, mental health symptoms, and health-related cognitions (risk perceptions, control beliefs). To separate between- and temporal within-person associations between mental health, health-related cognitions, and NPI adherence. Six-month ecological momentary assessment (EMA) study with six 4-day assessment bouts in 397 German adults. Daily measurement of adherence, mental health symptoms, and cognitions during bouts. We used dynamic temporal network analysis to estimate between-person, as well as contemporaneous and lagged within-person effects for distancing and hygiene NPIs. Distinct network clusters of mental health, health cognitions, and adherence emerged. Participants with higher control beliefs and higher susceptibility were also more adherent (between-person perspective). Within-person, similar findings emerged, additionally, distancing and loneliness were associated. Lagged findings suggest that better adherence to NPIs was associated with better mental health on subsequent days, whereas higher loneliness was associated with better subsequent hygiene adherence. Findings suggest no negative impact of NPI adherence on mental health or vice versa, but instead suggest that adherence might improve mental health symptoms. Control beliefs and risk perceptions are important covariates of adherence-both on between-person and within-person level.


Psychiatric referrals associated with substance use disorders: prevalence and gender differences. European Consultation-Liaison Workgroup.
A psychoactive substance use disorder (ICD-10) was diagnosed in 28% of 1222 psychiatric referrals in six general hospitals in Finland. In the age group 35 o 50 years, 53% of men and 29% of women had a substance use disorder. In the age group 35 to 50 years, sociodemographic characteristics differentiated men diagnosed with substance use disorders from other male consultation patients, but women with substance use disorders were similar to other female consultation patients. Urgency (i.e., consultation required within the same day) was accentuated in referrals associated with substance use disorders. Of attempted suicides, 65% were related to substance use disorders. Use of sedatives or hypnotics had more frequently induced disorders in women with substance use diagnosis compared to men with diagnosis (30% vs. 13%; p < 0.01). Female consultation patients with substance use disorders more often than respective male patients were current mental health outpatients. In conclusion, the proportion of substance use disorders among psychiatric consultation patients was remarkably higher in the present study (28%) compared with the average provided by the earlier literature (12%), and therefore, at least in Finland, psychiatric assessment of general hospital patients should always include assessment for the presence of substance use disorders. If substance use is only looked for in patients who have social problems typical of advanced misuse, detecting female substance use might be impeded. Relation of attempted suicides to substance use disorders was confirmed. To prevent misuse of prescribed drugs detected particularly in female consultation patients, and to prevent attempted suicides, doctors' attention is called to prescriptions of sedatives and hypnotics.