Assessment of Two Different Drinking Water Treatment Plants for the Removal of Free-living Amoebae, Egypt.
The aim of this study was to compare between slow and rapid sand filters for the removal of free-living amoebae during drinking water treatment production. Overall, 48 water samples were collected from two drinking water treatment plants having two different filtration systems (slow and rapid sand filters) and from inlet and outlet of each plant. Water samples were collected from Fayoum Drinking Water and Wastewater Holding Company, Egypt, during the year 2015. They were processed for detection of FLAs using non-nutrient agar (NNA). The isolates of FLAs were microscopically identified to the genus level based on the morphologic criteria and molecularly confirmed by the aid of PCR using genus-specific primers. The percentage of removal for FLAs through different treatment processes reached its highest rate in the station using slow sand filters (83%), while the removal by rapid sand filter system was 71.4%. Statistically, there was no significant difference (P=0.55) for the removal of FLAs between the two different drinking water treatment systems. Statistically, seasons had no significant effect on the prevalence of FLAs in the two different drinking water treatment plants. Morphological identification of the isolated FLAs showed the presence of 3 genera namely Acanthamoeba, Naegleria, and Vermamoeba (Hartmannella) confirmed by PCR. The appearance of FLAs especially pathogenic amoebae in completely treated drinking water may cause potential health threat although there is no statistical difference between the two examined drinking water filtration systems.


Clinico-pathological features of breast carcinoma in elderly Egyptian patients: a comparison with the non-elderly using population-based data.
Breast cancer (BC) is a major worldwide health care problem that mostly afflicts the elderly population in the more developed countries. It is not known how common is breast cancer among elderly Egyptian patients and whether this differs from the disease in younger patients. To study the clinico-pathological features of BC in elderly Egyptian patients (⩾65years of age) among the population of an Egyptian Governorate, Gharbiah, and to compare these features with those of younger patients (<65years). This is a cross sectional study that compares elderly BC (EBC) and the non-elderly BC (NEBC) using the information from the Gharbiah Population-based Cancer registry (GPCR) during the years 1999-2007. Out of 6078 BCs, 12% were EBCs and 88% were NEBCs. Between 1999 and 2007, the crude incidence rate (CIR, per 100,000 populations) of EBC increased from 47 to 71 and that of NEBC increased from 16 to 17. Compared to NEBC patients, EBC patients were more likely to have a positive family history and present with a distant disease and less likely to present with a localized disease. EBCs were more likely to have lung metastases and less likely to have liver metastases. Histology, grade, hormone and HER-2 receptor statuses were comparable in both groups. Apart from hormonal therapies, the elderly were less likely to receive surgery, radiotherapy or chemotherapy. EBC patients in Egypt present with advanced disease and are less likely to receive surgery, radiotherapy or chemotherapy compared to NEBC patients.


Extent of resection influences outcomes for patients with gliomas.
In recent years, advances in our understanding of the biology of low-grade gliomas (LGG) and high-grade gliomas (HGG) have driven new paradigms in molecular markers, diagnostic imaging, operative techniques and technologies, and adjuvant therapies. Taken together, these developments are collectively pushing the envelope towards improved quality of life and survival. Here, we review the recent literature to synthesize a comprehensive review of the value of extent of resection for LGGs and HGGs in the modern neurosurgical era.


TRIM35-mediated degradation of nuclear PKM2 destabilizes GATA4/6 and induces P53 in cardiomyocytes to promote heart failure.
Pyruvate kinase M2 (PKM2) is a glycolytic enzyme that translocates to the nucleus to regulate transcription factors in different tissues or pathologic states. Although studied extensively in cancer, its biological role in the heart remains unresolved. PKM1 is more abundant than the PKM2 isoform in cardiomyocytes, and thus, we speculated that PKM2 is not genetically redundant to PKM1 and may be critical in regulating cardiomyocyte-specific transcription factors important for cardiac survival. Here, we showed that nuclear PKM2 (S37P-PKM2) in cardiomyocytes interacts with prosurvival and proapoptotic transcription factors, including GATA4, GATA6, and P53. Cardiomyocyte-specific PKM2-deficient mice (Pkm2 Mut Cre+) developed age-dependent dilated cardiac dysfunction and had decreased amounts of GATA4 and GATA6 (GATA4/6) but increased amounts of P53 compared to Control Cre+ hearts. Nuclear PKM2 prevented caspase-1-dependent cleavage and degradation of GATA4/6 while also providing a molecular platform for MDM2-mediated reduction of P53. In a preclinical heart failure mouse model, nuclear PKM2 and GATA4/6 were decreased, whereas P53 was increased in cardiomyocytes. Loss of nuclear PKM2 was ubiquitination dependent and associated with the induction of the E3 ubiquitin ligase TRIM35. In mice, cardiomyocyte-specific TRIM35 overexpression resulted in decreased S37P-PKM2 and GATA4/6 along with increased P53 in cardiomyocytes compared to littermate controls and similar cardiac dysfunction to Pkm2 Mut Cre+ mice. In patients with dilated left ventricles, increase in TRIM35 was associated with decreased S37P-PKM2 and GATA4/6 and increased P53. This study supports a previously unrecognized role for PKM2 as a molecular platform that mediates cell signaling events essential for cardiac survival.


Catecholamines and attention. I: Animal and clinical studies.
One important function of the catecholamine innervation of the cerebral cortex may be the control of attention. Of particular interest are the catecholamine projections to the cerebral cortex from the reticular formation, namely the dopamine neurons of the ventral tegmentum of the midbrain and the noradrenergic neurons of the locus coeruleus in the upper pons. Animal studies implicate noradrenaline and dopamine in a wide range of attention-related behaviours involving search and exploratory activity, distractibility, response rate, discriminability and the switching of attention. Most human studies come from the clinical literature relating to schizophrenia, Parkinson's disease and attention deficit disorder. An association has been claimed in each of these conditions between abnormal catecholamine activity (in particular dopamine) and attentional dysfunction. In particular, difficulty with the attachment of appropriate responses to environmental stimuli, akin to those observed in animals with lesions to central dopamine pathways, indicates a role for dopamine in response selection processes. Overall, the animal and human studies reviewed indicate a role for central noradrenaline and dopamine in the early and late processing of information, respectively.


The temporal course of COVID-19 anosmia and relation to other clinical symptoms.
This study aimed to define the clinical course of anosmia in relation to other clinical symptoms. 135 patients with COVID-19 were reached by phone and subsequently included in the study. Olfactory functions were evaluated using a questionnaire for assessment of self-reported olfactory function. Patients were divided into four subgroups according to the presence of olfactory symptoms and temporal relationship with the other symptoms: group1 had only olfactory complaints (isolated, sudden-onset loss of smell); group2 had sudden-onset loss of smell, followed by COVID-19 related complaints; group3 initially had COVID-19 related complaints, then gradually developed olfactory complaints; and group4 had no olfactory complaints. In total, 59.3% of the patients interviewed had olfactory complaints during the disease course. The olfactory dysfunction severity during COVID-19 infection was significantly higher in group1 compared to groups 2 and 3. In groups1-3, the odor scores after recovery from COVID-19 disease were significantly lower compared to the status prior to disease onset. The residual olfactory dysfunction was similar between groups1 and 2, but was more evident than group3. Mean duration for loss of smell was 7.8 ± 3.1 (2-15) days. Duration of loss of smell was longer in groups1 and 2 than in group3. Odor scores completely returned back to the pre-disease values in 41 (51.2%) patients with olfactory dysfunction. Rate of complete olfactory dysfunction recovery was higher in group3 compared to groups1 and 2. In isolated anosmia cases, anosmia is more severe, and complete recovery rates are lower compared to the patients who have other clinical symptoms. Level 4.


Dried blood spot sampling for detection of monoclonal immunoglobulin gene rearrangement.
Molecular methods are important tools for diagnosis and monitoring of many lymphoproliferative disorders. The reliability of lymphoma diagnoses is strikingly different between developed and developing countries, partly due to lack of access to these advanced molecular analyses. To overcome these problems, we propose a new application of dried blood spots (DBS) for detecting clonal B-cell populations in peripheral blood (PB). We ensured that the DBS contained sufficient lymphocytes to perform a PCR-based clonality assay without producing false positives. Using the Namalwa B-cell line, we established that the assay is sensitive enough to detect 200 clonal cells in the analyzed sample. Very similar clonal results were obtained between DNA from DBS and fresh whole blood from patients with B-cell chronic lymphocytic leukemia. B-cell clonality can also be detected in DBS from African children with EBV-associated diseases. This is the first study demonstrating that clonality testing can be performed on DBS samples, thus improving the diagnostic and monitoring options for lymphoproliferative diseases in resource-limited settings.


Toll‑like receptor 4 activates the NLRP3 inflammasome pathway and periodontal inflammaging by inhibiting Bmi‑1 expression.
Overproduction of pro‑inflammatory cytokines in the aged, which is called inflammaging, leads to the deterioration of periodontitis. Toll‑like receptor 4 (TLR4) plays a role in the regulation of cellular senescence, and its expression increases with age. However, there has been limited research into the molecular mechanisms underlying the onset of periodontal inflammaging, and the interplay between TLR4 and inflammaging. In the present study, wild‑type and TLR4 gene knockout mice were used to investigate the activation of the TLR4 pathway in mouse periodontitis and the expression of the nucleotide‑binding and oligomerization domain‑like receptor 3 (NLRP3) inflammasome, an upstream immune checkpoint during the development of inflammaging. Activation of TLR4 in a mouse model of periodontitis enhanced the expression of a senescence‑associated secretory phenotype (SASP), which boosted the inflammaging process. Conversely, TLR4 activation downregulated the expression of B cell‑specific Moloney murine leukemia virus integration site 1 (Bmi‑1) and promoted the priming of NLRP3 inflammasome, both of which are regulators of SASP. Treating gingival fibroblasts with Bmi‑1 inhibitor PTC209, it was demonstrated that TLR4 activated the NLRP3 pathway and the inflammaging process by suppressing Bmi‑1. In addition, there was a significant reduction in the expression of Bmi‑1 expression in the gingiva of patients with periodontitis compared with healthy controls. In conclusion, the present study demonstrated that TLR4 acted by inhibiting Bmi‑1 to enhance the NLRP3 pathway and SASP factors. This cascade of reactions may contribute to the senescence of the periodontium.


Selective oviposition by a leaf miner in response to temporal variation in abscission.
We studied within-tree variaton in density and mortality of the leaf miner, Cameraria sp. nov., on Emory oak. Miner densities were lower in the peripheral than the central parts of trees. Following tagged mined and unmined leaves revealed that peripheral leaves fall sooner than central leaves and that more Cameraria mining peripheral leaves die due to abscission than those mining central leaves. We discuss factors that might influence interactions between leaf miners and abscission of the host-leaves they attack.


Crystallization and preliminary crystallographic studies of Hyp-1, a St John's wort protein implicated in the biosynthesis of hypericin.
According to a debated hypothesis, the biosynthesis from emodin of the medicinally important natural compound hypericin is catalyzed in St John's wort (Hypericum perforatum) by the phenolic oxidative-coupling protein Hyp-1. Recombinant St John's wort Hyp-1 has been overexpressed in Escherichia coli and obtained in single-crystal form. The crystals belong to the orthorhombic system, space group P2(1)2(1)2(1), with unit-cell parameters a = 37.5, b = 76.7, c = 119.8 A, contain two protein molecules in the asymmetric unit and diffract X-rays to 1.73 A resolution.


Design, Assembly, and Function of DNA Origami Mechanisms.
This chapter provides an overview of the common procedures used in making functional DNA origami devices. These procedures include the design, assembly, purification, and characterization of the DNA origami structures, with a focus on dynamic devices.


Leptospirosis in Colombia: isolation of Leptospira spp. from the kidneys of brown rats (Rattus norvegicus) trapped on infected piggeries.
Leptospira interrogans serotypes pomona and icterohaemorrhagiae were isolated from the kidneys of Rattus norvegicus trapped on pomona infected piggeries. The isolation of pomona from one of 111 cultured kidneys, although of interest, strongly suggests that this rat does not play an important part in the epidemiology of infection by this serotype. Kidney tubular degeneration, focal mononuclear infiltrations and fibroplasia are morphological traces which remain after an attack of leptospirosis. The high percentage (48.6) of these lesions found in the rat kidneys examined does not seem to be related to serotype pomona but to icterohaemorrhagiae. The importance of this last serotype in swine in the Cauca Valley has yet to be assessed.


The combined contraceptive vaginal device (NuvaRing): a comprehensive review.
The aim of this study is to review the development of NuvaRing over the past decade to illustrate its use-effectiveness and acceptance as an alternative contraceptive option for women. The data were extracted from the literature using computerised MEDLINE system. NuvaRing is a new combined hormonal contraceptive vaginal ring made of ethylene-vinyl-acetate copolymer, releasing 120 microg etonorgestrel and 15 microg ethinyloestradiol per day. This ring is inserted on any day from day 1 to day 5 of a menstrual cycle for 21 days, thereafter removed for 7 days ring-free period and discarded. Complete inhibition of ovulation is observed during treatment with this device. Clinical exposure to NuvaRing for 1786 women-years has resulted in 21 pregnancies, giving a Pearl Index of 1.18. Withdrawal bleeding (4.7-5.3 days) is regular (97-99% of cycles) with rare incidence of irregular bleeding (2.6-6.4%). The cycle control is good with the use of this combined contraceptive vaginal ring. NuvaRing is well tolerated and accepted by women as compared to oral pill. NuvaRing is an effective vaginal contraceptive option for women. However, further study is needed for monitoring its long-term effectiveness and impact on patient's quality of life since the NuvaRing is marketed in many countries.


Postoperative information needs and communication barriers of esophageal cancer patients.
Given the poor prognosis of esophageal cancer and the impact of surgery on health-related quality of life (HRQL), addressing patients' postoperative information needs is important. This study aimed to examine (1) the content and type of patients' information needs and (2) patient perceived facilitators and barriers to patient participation. Interviews were conducted with 20 purposefully selected esophageal cancer patients. Open and structured questions were alternated. The transcribed interviews were analysed inductively and deductively, using MAXqda. Patients' post-operative information needs concerned HRQL, medical care and prognosis, covering several sub-domains. Different types of needs were identified, e.g., requests for information about cause, course and self-management. Barriers to patient participation mostly reflected beliefs and skills, and could be categorized into agenda and communication barriers. Facilitators of patient participation reflected physician, patient and interaction characteristics, companion support and pre-visit preparation. Many patients saw merit in pre-visit preparation interventions; few endorsed skill-building interventions. This study listed the postoperative information needs of esophageal cancer patients. Moreover, it gained insight into patient-perceived barriers and facilitators of patient participation. The findings demonstrate what information physicians should have available and informs interventions to support patients in meeting their information needs.


Upregulation of histamine receptor H1 promotes tumor progression and contributes to poor prognosis in hepatocellular carcinoma.
H1 histamine receptor (H1HR) belongs to the family of rhodopsin-like G-protein-coupled receptors. Recent studies have shown that H1HR expression is increased in several types of cancer. However, its functional roles in tumor progression remain largely unknown, especially in hepatocellular carcinoma (HCC). We found that H1HR is frequently unregulated in HCC, which is significantly associated with both recurrence-free survival and overall survival in HCC patients. Functional experiments revealed that H1HR promoted both the growth and metastasis of HCC cells by inducing cell cycle progression, formation of lamellipodia, production of matrix metalloproteinase 2, and suppression of cell apoptosis. Activation of cyclic adenosine monophosphate-dependent protein kinase A was found to be involved in H1HR-mediated HCC cell growth and metastasis. In addition, we found that overexpression of H1HR was mainly due to the downregulation of miR-940 in HCC cells. Moreover, the H1HR inhibitor terfenadine significantly suppressed tumor growth and metastasis in an HCC xenograft nude mice model. Our findings demonstrate that H1HR plays a critical role in the growth and metastasis of HCC cells, which provides experimental evidence supporting H1HR as a potential drug target for the treatment of HCC.


Appraisal of the DIERS method for calculating postural measurements: an observational study.
Surface topography is increasingly used with postural analysis. One system, DIERS formetric 4D, measures 40 defined spine shape parameters from a 6-s scan. Through system algorithms, a set of spine shape parameter values from 1 of 12 recorded images obtained during a scan becomes the DIERS-reported value (DRV) for postural assessment. The purpose of the current study was to compare DRV with a standard average value (SAV) calculated from all 12 images to determine which method is more appropriate for assessing postural change. One mannequin and 30 human participants were scanned over 5 days. Values from each image and the DRV for 40 defined spine shape parameters were exported, and mean DRV, mean SAV, mean DRV, and within-scan variance were calculated. Absolute difference and percent change between mean DRV and mean SAV were calculated for the mannequin and humans. Inter-method reliability was calculated for humans. Within-scan variance for each parameter was tested for significant variability. For all spine shape parameters on the mannequin, absolute difference (< 0.6 mm, 0.1°, or 0.1%) and percent change (< 2.90%) between mean DRV and mean SAV for each parameter were small. Nine parameters on human participants had a large percent change (> 7%). Absolute difference between mean DRV and mean SAV for those nine parameters was small (≤ 0.87 mm or 0.61°). Absolute difference for all other parameters ranged from 0.02 to 6.98 mm for distance measurements, from 0.01 to 1.21° for angle measurements, and from 0.15 to 0.22% for percentage measurements. Inter-method reliability between DRV and SAV was excellent (0.94-1.00). For the mannequin, within-scan variance was small (< 1.62) for all parameters. For humans, within-scan variance ranged from 0.05 to 36.04 and was different from zero for all parameters (all P < 0.001). The minimal variability observed in the mannequin suggested the DIERS formetric 4D instrument had high within-scan reliability. The DRV and SAV provided comparable spine shape parameter values. Because within-scan variability is not reported with the DRV, the clinical usefulness of current DRV values is limited. Establishing an estimate of variance with the SAV will allow clinicians to better identify a clinically meaningful change.


Effect of thyroid hormone on hepatic cholesterol 7 alpha hydroxylase, LDL receptor, HMG-CoA reductase, farnesyl pyrophosphate synthetase and apolipoprotein A-I mRNA levels in hypophysectomized rats.
The effects of thyroid hormone on cholesterol 7 alpha hydroxylase, LDL receptor, HMG-CoA reductase, apo A-I and farnesyl pyrophosphate synthetase hepatic mRNA levels were investigated in hypophysectomized rats. Of these mRNAs cholesterol 7 alpha hydroxylase responded the most rapidly and required the lowest dose of T3. Maximal mRNA levels were reached one hr after T3 administration and required 10 micrograms/100g of body weight. These results suggest that the hypocholesterolemic effect of thyroid hormone may be mediated by a primary effect on cholesterol 7 alpha hydroxylase gene expression.


Thyroid Peroxidase Antibodies in Non-Interferon Treated Hepatitis C Patients in Pakistan.
Interferon therapy of HCV infected patients is associated with development of thyroid dysfunctions. Patients with pretreatment presence of antithyroid peroxidase (TPO-Ab) are at greater risk. This study, probably the first in Pakistan, was planned to determine TPO-Ab in sera of treatment-naive local HCV patients. Setting. Centre for Nuclear Medicine (CENUM), Mayo Hospital, Lahore. During July to December 2012, 190 patients (140 females, 50 males) newly diagnosed for HCV infection were selected for this study. Their age range was 15-55 years (mean: 35.3 ± 9.1 years). 262 age matched healthy subjects (211 females and 50 males) were recruited as control. Serum-free thyroxin (FT4) and thyroid stimulating hormone (TSH) were detected by radioimmunoassay techniques. Serum TPO-Ab titer was determined by ELISA method using commercial kits. Serum FT4 and TSH levels in HCV patients and controls were within normal range. Between two groups there was no significant difference in mean value of FT4 (16.0 ± 3.0 versus 16.2 ± 3.9; P = 0.619) but mean TSH value was significantly lower in HCV patients (1.5 ± 0.8 versus 1.8 ± 0.9; P = 0.003). Among HCV patients 51 (26.8%) were TPO-Ab positive and among control subjects 18 (6.9%) were TPO-Ab positive. The difference was statistically significant (P < 0.001). Further analysis showed that among HCV patients 39 (27.8%) females and 12 (24.0%) males were TPO-Ab positive, respectively, and difference was not statistically significant (P = 0.873). Moreover, TPO-Ab positive patients were older and had significantly higher serum TSH as compared to TPO-Ab negative HCV patients. Independent of patient's gender and increasing with advancing age, about one-fourth of local untreated HCV patients are TPO-Ab positive and are at greater risk of developing thyroid disorders during and after interferon treatment.


Serum lipids, plant sterols, and cholesterol kinetic responses to plant sterol supplementation in phytosterolemia heterozygotes and control individuals.
Plant sterol (PS) supplementation is increasingly accepted as a dietary strategy to lower plasma cholesterol concentrations. However, information is scarce about the effect of increased PS intake in potentially vulnerable groups, such as phytosterolemia heterozygotes (HET). This study assessed the responsiveness of circulating PS and lipid concentrations and cholesterol kinetics (absorption and synthesis) to daily PS supplementation in HET (ABCG8 S107X mutation) compared with a healthy control cohort. A double-blind, randomized, crossover, placebo-controlled study was conducted in 10 HET and 15 control subjects. The participants had a mean (±SEM) age of 34 ± 2 y and a BMI (in kg/m²) of 29.9 ± 1.1 and consumed ∼1.6 g PS or placebo capsules daily with supper for 4 wk. Cholesterol absorption and synthesis were assessed by using [¹³C]cholesterol and deuterium oxide, respectively. Plasma LDL-cholesterol concentrations decreased (P = 0.006) in both groups after PS supplementation (HET: 2.73 ± 0.19 mmol/L; control: 3.11 ± 0.19 mmol/L) compared with placebo (HET: 3.12 ± 0.20 mmol/L; control: 3.50 ± 0.21 mmol/L), whereas PS concentrations (campesterol+β-sitosterol) increased (P = 0.03) in both groups after PS supplementation (HET: 39.72 ± 6.05 μmol/L; control: 24.03 ± 1.65 μmol/L) compared with placebo (HET: 27.32 ± 3.80 μmol/L; control: 21.12 ± 2.05 μmol/L). Cholesterol absorption efficiency decreased (P = 0.010) by ∼22% and ∼17% and synthesis rates increased (P = 0.040) by ∼20% and ∼24% in the HET and control groups, respectively, in response to PS consumption compared with placebo. These data suggest that heterozygosity for the ABCG8 S107X mutation does not influence the action of dietary PS on circulating cholesterol concentrations but may affect sterol absorption.


Sphingomonas pokkalii sp. nov., a novel plant associated rhizobacterium isolated from a saline tolerant pokkali rice and its draft genome analysis.
Three strains L3B27T, 3CNBAF, L1A4 isolated from a brackish cultivated pokkali rice rhizosphere were characterised using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA and recA gene sequences revealed that these strains were highly similar among each other and formed a separate monophyletic cluster within the genus Sphingomonas with Sphingomonas pituitosa DSM 13101T, Sphingomonas azotifigens DSM 18530T and Sphingomonas trueperi DSM 7225T as their closest relatives sharing 97.9-98.3% 16S rRNA similarity and 91.3-94.0% recA similarity values, respectively. The average nucleotide identity (ANI), average amino acid identity (AAI) and digital DNA-DNA hybridisation (dDDH) values between L3B27T (representative of the novel strains) and its phylogenetically closest Sphingomonas species were well below the established cut-off <94% (ANI/AAI) and <70% (dDDH) for species delineation. Further, the novel strains can be distinguished from its closest relatives based on several phenotypic traits. Thus, based on the polyphasic approach, we describe a novel Sphingomonas species for which the name Sphingomonas pokkalii sp. nov (type strain L3B27T=KCTC 42098T=MCC 3001T) is proposed. In addition, the novel strains were characterised for their plant associated properties and found to possess several phenotypic traits which probably explain its plant associated lifestyle. This was further confirmed by the presence of several plant associated gene features in the genome of L3B27T. Also, we could identify gene features which may likely involve in brackish water adaptation. Thus, this study provides first insights into the plant associated lifestyle, genome and taxonomy of a novel brackish adapted plant associated Sphingomonas.


Management of the Nasal Dorsum: Construction and Maintenance of a Barrel Vault.
Rhinoplasty is not so much an art, but rather an architectural undertaking: a methodical approach to reconfiguring the nasal components to give a proportionate nose that both pleases the eye and satisfies functional requirements. The dorsum and dorsal esthetic lines are some of the most important components of the nose in terms of esthetics and function. The middle vault is the critical portion of the nose that will guide the management of the bony vault and the tip. The role of the spreader flap and its extension into the bony vault is stressed to re-create the barrel vaultlike nasal architecture.


Butyrylcholinesterase in pericytes associated with canine brain capillaries.
The distribution of the enzyme butyrylcholinesterase (BChE) in dog brain cortex and cerebellum was studied by light and electron microscopy using a Hatchett's brown technique. Staining was found in neuron cell bodies and processes, the white matter of the cerebellum, and in capillaries and arterioles. Electron microscopy indicated that the enzyme activity associated with vessels was present in pericytes. Reaction product was found at the cell membrane, the intermembranous space of the nuclear envelope, and the Golgi complex of these cells. The finding of BChE in canine brain pericytes and its absence in endothelium does not support the idea that this enzyme is important in blood-brain barrier function. The pericyte in dog brain may be a site of synthesis of this enzyme and is, in this respect, similar to the endothelial cell of rat brain.


The relationship between features of residency training and ABIM certifying examination performance.
The purpose of this study was to provide data concerning the relationship between features of residency training and a test of cognitive achievement gathered at the end of residency. To accomplish this, data collected in the late 1970s by three national organizations were joined and analyzed with the aid of experts in internal medicine. Although graduate medical education has evolved since this information was gathered, it does provide a baseline for assessing the impact of changes on the cognitive skills of residents. The findings suggest that better program performance on the examination is associated with attracting more knowledgeable residents to begin with and that programs are able to maintain the advantage of their residents throughout training. Moreover, program characteristics have an impact on the cognitive skills of residents over and above what would be predicted by test scores at the end of medical school. Programs with better examination performance tend to provide residents an extensive, well-supervised educational experience stressing ambulatory care.


Management of chronic coronary disease: is the pendulum returning to equipoise?
Over the last 3 decades, our ability to mechanically dilate obstructive coronary arterial stenoses has fundamentally altered our approach to managing patients with coronary artery disease (CAD). The result has been a swing from an initial pharmacologic approach to anatomically driven revascularization. An accumulation of clinical evidence provides strong support for such intervention in acute coronary syndromes (ACS). In stable CAD, dilative therapy was believed to be superior based on the assumption that high-risk coronary anatomy or myocardial ischemia increases the risk of future death and myocardial infarction. However, there have been major advances in our understanding of the pathophysiology of ACS and the recognition of the significance of predisposing non-flow-limiting coronary stenoses prone to rupture, as well as increasing insight into plaque and patient vulnerability. This improved understanding of the disease has led to the more aggressive use of appropriately targeted pharmacologic agents and an evolution in what constitutes optimal medical therapy (OMT). Data from recent studies, such as the Clinical Outcomes Utilizing Revascularization and Aggressive Drug Evaluation (COURAGE) trial, support the concept that in patients with stable CAD, OMT alone in this day and age compares favorably with a therapeutic strategy combining OMT with mechanical intervention. Thus, the treatment pendulum may be swinging back to the understanding that "best practice" today requires the judicious use of interventional and medical therapies in the appropriate patient population.


Faculty perceptions of effective ambulatory care teaching.
The authors interviewed faculty members to determine their perceptions of what constitutes effective teaching in the ambulatory setting. They conducted semistructured interviews with experienced clinician-tutors who supervise residents in two internal medicine clinics. Tutors identified similarities as well as important differences between inpatient teaching and outpatient teaching. Questioning, role modeling, and emphasizing general principles and concept comprehension can be used effectively in both settings. On the other hand, the two settings differ strikingly in teaching of problem solving, bedside teaching, and provision of feedback. Many characteristics of the setting influence outpatient teaching, but the tutors offered differing viewpoints about whether these characteristics are beneficial or detrimental.


Cognitive Processes and Translation Quality: Evidence from Keystroke-Logging Software.
This study investigates the relationship between cognitive processes and translation quality in the context of English-Arabic translations of journalistic articles. Specifically, it explores the translation processes at the orientation, production, and revision stages and the relationship between such processing and translation quality, using keylogging software (i.e., Translog II) to record the experiment. Twenty-two translation trainees participated in the study, translating a news article from English into Arabic. Presas's (2012) rubric for assessment was used to evaluate the translation quality, and several correlation analyses were applied to the data. Findings revealed negative correlations between translation quality and online revision, translation duration, and text production. The trainees' translations demonstrated limitations in communicating the main ideas of the target text (TT) to the target language (TL). The findings also showed the trainees' focus on online revision and editing and the concentration of translation time and cognitive effort in the drafting phase. The researchers recommend including the three phases of translation (reading, drafting, and revision) into translation training courses and equipping translator trainees with the required skills for each translation stage.


Separation of human milk oligosaccharides by recycling chromatography. First isolation of lacto-N-neo-difucohexaose II and 3'-Galactosyllactose from this source.
Lacto-N-neo-difucohexaose II, beta-D-Galp-(1----4)-[alpha-L-Fucp-(1----3)]-beta-D-Glcp-NAc-(1----3)-beta-D-Galp-(1----4)-[alpha-L-Fucp-(1----3)]-D-Glc, and 3'-galactosyllactose, beta-D-Galp-(1----3)-beta-D-Galp-(1----4)-D-Glc, were isolated for the first time from human milk by means of a recycling chromatography technique. Through this method, carried out mainly on columns of K+ ion-exchange resins and either Bio-Gel P-4 or TSK 40 W(S) gel filtration media, up to one gram of an oligosaccharide mixture could be handled and lacto-N-neo-difucohexaose II separated from isomeric lacto-N-difucohexaose I, alpha-L-Fucp-(1----2)-beta-D-Galp-(1----3)-[alpha-L-Fucp-(1----4)]-beta-D-GlcpNAc-(1----3)-beta-D-Galp-(1----4)-D-Glc, and II, beta-D-Galp-(1----3)-[alpha L-Fucp-(1----4)]-beta-D-GlcpNAc-(1----3)-beta-D-Galp-(1----4)-[alpha-L-Fucp-(1----3)]-D-Glc. This method also permitted resolution of isomeric mixtures of the trisaccharides 2'-fucosyllactose, alpha-L-Fucp-(1----2)-beta-D-Galp-(1----4)-D-Glc, and 3-fucosyllactose, beta-D-Galp-(1----4)-[alpha-L-Fucp-(1----3)]-D-Glc, the tetrasaccharides lacto-N-tetraose, beta-D-Galp-(1----3)-beta-D-GlcpNac-(1----3)-beta-D-Galp-(1----4)-D-Glc, and lacto-N-neo-tetraose, beta-D-Galp-(1----4)-beta-D-GlcpNAc-(1----3)-beta-D-Galp-(1----4)-D-Glc, and the pentaoses lacto-N-fucopentaose I, alpha-L-Fucp-(1----2)-beta-D-Galp-(1----3)-beta-D-GlcpNAc-(1----3)-beta-D-Galp-(1----4)-D-Glc, II, beta-D-Galp-(1----3)-[alpha-L-Fucp-(1----4)]-beta-D-GlcpNAc-(1----3)-beta-D-Galp-(1----4)-D-Glc, and III, beta-D-Galp-(1----4)-[alpha-L-Fucp-(1----3)]-beta-D-GlcpNAc-(1----3)-beta-D-Galp-(1----4)-D-Glc, which have proved difficult if not impossible to separate by other means. The isolation of these and other milk oligosaccharides is described herein. The 500-MHz 1H-n.m.r. spectra of lacto-N-neo-difucohexaose II and 3'-galactosyllactose, and their alditols, are recorded. 1H-n.m.r. data on some other milk oligosaccharides, both natural and reduced, are also given.


Effect of amaranth and quinoa seed flour on rheological and physicochemical properties of goat meat nuggets.
An attempt was made through the present study to prepare gluten free goat meat nuggets by replacing refined wheat flour from product formulation with healthy, dietary fibre rich amaranth (A) and quinoa (Q) flour at different levels. A total of five different treatments viz., AI (1.5% amaranth), AII (3% amaranth), QI (1.5% quinoa) and QII (3% quinoa) were prepared. The physicochemical, colour, texture, sensory and rheological properties of these pseudocereal-added products were evaluated against control (3% refined wheat flour). Emulsion stability of meat batter was significantly affected (P < 0.05) due to addition of amaranth flour (1.5% and 3%) and quinoa (3%). Treatment groups AII and QI had significantly low (P < 0.05) moisture content with respect to control while the amount of fat content showed a reverse trend. Addition of amaranth and quinoa significantly increased (P < 0.05) the dietary fibre in meat products. Rheology of meat batter was affected by types and level of pseudocereal incorporation. Treatment AII had low (P < 0.05) hunter colour lightness value, whereas redness value was low for treatment QI. Effect of added amaranth and quinoa flour was observed on the textural parameters like adhesiveness, cohesiveness, gumminess and chewiness. High scores for almost all the sensory parameters were recorded in pseudocereal-added meat products. Though, all the products were very much acceptable, product with 1.5% quinoa flour was found to have high (P < 0.05) overall acceptability score.


The analysis of multiple habituation profiles of single trial evoked potentials.
A new statistical technique for the analysis of multiple habituation profiles of single trial evoked potentials is presented. The method is based on a model that highlights the essential characteristics of the habituation process, viz. trial-dependent modulation of amplitude and phase of the underlying brain responses to a repeated stimulus, while recognizing the presence of unsystematic background variation inherent to single trial registrations. Without the introduction of any simplifying assumptions about the time course of habituation, a set of single trial evoked potentials is decomposed into one or more underlying evoked potentials, their habituation profiles across trials and the corresponding true evoked potentials. The analysis of the model proceeds by means of spectral analysis and its validity is illustrated with applications to simulated data.


[Significance of tube manometry in the assessment of tube function. A comparison of tube manometry results and continuous direct determination of pressure in the middle ear].
Tube manometry--also known as aspiration-deflation-test--is an often used, practicable clinical test for tube function. By comparing tube manometric results with continual measurements of the middle ear pressure, a close correlation between both test results could be demonstrated. Tube manometric tests thus permit an estimation of the tube function as well as of the probable development of the middle ear pressure. An exact classification in terms of physiologic and pathologic tube function, however, is not possible. It is clear that even a partial equalization of low-pressure values in tube manometry should be interpreted as physiologic.


Vasodilator effect of terbutaline: in vivo evidence for the existence of beta 2-adrenoceptors in the microcirculation of rats, hamsters, and guinea pigs.
The characteristics of beta 2-adrenoceptors were studied in vivo in the microcirculation of cremaster muscle and mesoappendix in rats, cheek pouch in hamsters, and mesentery in guinea pigs, using a video-microscope technique. Terbutaline (beta 2-selective agonist) caused vasodilatation in all four microcirculation preparations studied, whereas butoxamine (beta 2-selective antagonist) competitively inhibited the terbutaline-induced vasodilatation. The in vivo pA2 values of butoxamine inhibition of terbutaline vasodilator effect were estimated from Schild plots. The pA2 values (6.22-6.31) were statistically identical in all four microcirculations studied and similar to in vitro values reported in other tissues. In contrast, tazolol and prenalterol (both beta 1-selective agonists) did not produce any vasodilatation in the microcirculations studied. These results supported the existence of beta 2-adrenoceptors involved in vasodilatation in the microcirculation.


(-)-Epigallocatechin gallate suppresses the cytotoxicity induced by trichothecene mycotoxins in mouse cultural macrophages.
Trichothecene mycotoxins are toxic secondary metabolites produced by a number of fungi including Fusarium species, which adversely affect lymphocytes. Deoxynivalenol (DON) and HT-2 toxin (HT-2) belong to the trichothecene group of mycotoxins and the occurrence of cereals and foodstuffs with these compounds are serious health problems. The aim of this study was to examine the effect of (-)-epigallocatechin gallate (EGCG), one of the main components in green tea catechins, on DON- or HT-2-induced cytotoxicity in mouse macrophages. EGCG had protective effects against the trichothecene-induced cytotoxicities of both mycotoxins. Additionally, EGCG suppressed the DON-induced activation of caspase-3/7, which is an indicator of apoptosis. These results indicate that EGCG might be useful in protection against DON- or HT-2-induced cell death, suggesting that EGCG could contribute to reducing the toxicities of trichothecenes.


Synthesis and Anti-Inflammatory Activity Evaluation of Some Benzimidazole Derivatives.
Hyper-inflammation aggravates the symptoms of both communicable and non-communicable diseases. Therefore, anti-inflammatory compounds may have wide therapeutic application. Benzimidazole is a privileged scaffold and its success in drug development is evident from the long list of benzimidazole-based drugs with wide range of applications. This study was undertaken to develop new small molecules with anti-inflammatory properties. Compounds MBPHYD, MBNHYD and MBHYDX were synthesised, purified, characterised and found to be non-toxic both in vitro (in 100 μMconcentration for 24 h vs. 3000 Vero cells/well) and in vivo (at a dose of 100 mg/kg in female Wistar rats with animals observed for 48 h for any mortality). Compounds MBPHYD and MBNHYD were found to possess significant anti-inflammatory properties. Further, in silico analysis suggested their compliance with drug-likeness. While no toxicity was predicted, both compounds were suggested to have good oral bioavailability. Thus, results of this study may encourage further investigation to establish new anti-inflammatory benzimidazoles for application against various disease conditions.


Kainic acid prevents peroxidase labeling of retinal ganglion cell bodies in the rat: a possible gate in retrograde axonal transport.
Kainic acid, a neurotoxic analogue of glutamate, injected into the vitreous body of the eye, interferes with the retrograde transport of the marker enzyme horseradish peroxidase. This interference is expressed by the absence of detectable peroxidase in the cell bodies of the retinal ganglion cells and by the presence of the marker in the intraretinal portions of the optic axons. These results suggest the hypothesis of the existence of some kind of gate at the proximal portion of the axons which would control the entry of retrogradely moving material into the cell body.


Noise Exposures Causing Hearing Loss Generate Proteotoxic Stress and Activate the Proteostasis Network.
Exposure to excessive sound causes noise-induced hearing loss through complex mechanisms and represents a common and unmet neurological condition. We investigate how noise insults affect the cochlea with proteomics and functional assays. Quantitative proteomics reveals that exposure to loud noise causes proteotoxicity. We identify and confirm hundreds of proteins that accumulate, including cytoskeletal proteins, and several nodes of the proteostasis network. Transcriptomic analysis reveals that a subset of the genes encoding these proteins also increases acutely after noise exposure, including numerous proteasome subunits. Global cochlear protein ubiquitylation levels build up after exposure to excess noise, and we map numerous posttranslationally modified lysines residues. Several collagen proteins decrease in abundance, and Col9a1 specifically localizes to pillar cell heads. After two weeks of recovery, the cochlea selectively elevates the abundance of the protein synthesis machinery. We report that overstimulation of the auditory system drives a robust cochlear proteotoxic stress response.


Two forms of HLA class I molecules in human plasma.
Soluble HLA-A,-B antigens have previously been detected in human plasma. More recently, these molecules have been demonstrated to be secreted in water soluble form by cell lines and peripheral blood lymphocytes in vitro due to RNA splicing events which delete exon five from a fraction of class I antigen transcripts. In order to determine whether plasma HLA-A,-B molecules arise by this mechanism, their biochemical properties are analyzed. Based upon molecular weight, detergent binding, and lipid binding properties, it was demonstrated that two forms of HLA-A,-B molecules are present in the plasma. A 43 kd form that binds both lipids and detergents is presumed to be a shed version of the membrane form. However, a 39 kd form that fails to bind lipids and detergents has properties identical to those molecules secreted in vitro. It is suggested that these molecules arise by alternative splicing events in vivo that are identical to those characterized in vitro. Although all tested individuals display shed and/or secreted plasma class I molecules, it is striking that HLA-A24 occurs in both a shed and a secreted form at levels higher than all other alleles examined.


RNA toxicity is a component of ataxin-3 degeneration in Drosophila.
Polyglutamine (polyQ) diseases are a class of dominantly inherited neurodegenerative disorders caused by the expansion of a CAG repeat encoding glutamine within the coding region of the respective genes. The molecular and cellular pathways underlying polyQ-induced neurodegeneration are the focus of much research, and it is widely considered that toxic activities of the protein, resulting from the abnormally long polyQ tract, cause pathogenesis. Here we provide evidence for a pathogenic role of the CAG repeat RNA in polyQ toxicity using Drosophila. In a Drosophila screen for modifiers of polyQ degeneration induced by the spinocerebellar ataxia type 3 (SCA3) protein ataxin-3, we isolated an upregulation allele of muscleblind (mbl), a gene implicated in the RNA toxicity of CUG expansion diseases. Further analysis indicated that there may be a toxic role of the RNA in polyQ-induced degeneration. We tested the role of the RNA by altering the CAG repeat sequence to an interrupted CAACAG repeat within the polyQ-encoding region; this dramatically mitigated toxicity. In addition, expression of an untranslated CAG repeat of pathogenic length conferred neuronal degeneration. These studies reveal a role for the RNA in polyQ toxicity, highlighting common components in RNA-based and polyQ-protein-based trinucleotide repeat expansion diseases.


The development of humoral immunity to tissue-specific tubular basement membrane alloantigens after renal transplantation across the major histocompatibility barrier in rats immunomodulated with blood transfusions and cyclosporin.
The development of a humoral immune response to the tubular basement membrane (TBM) alloantigen of Brown-Norway (BN) rat kidneys was studied after transplantation of BN rat kidneys into bilaterally nephrectomized Lewis (LEW) rats. The LEW rat recipients consisted of four groups receiving no form of immunosuppression, pretransplantation cyclosporin alone, or pretransplantation donor-specific or donor-nonspecific transfusions combined with cyclosporin. The latter two regimens induce indefinite allograft survival in the majority of recipients. Circulating antibody to collagenase-solubilized BN rat renal basement membrane (CS-BN-RBM) was present in all four groups of transplant recipients within 1 week after transplantation, and no significant differences in antibody levels were noted between rats receiving no immunosuppression (survival of 1-2 weeks) and the groups of rats who received various immunosuppressive regimens and survived longer. Circulating antibody to BN-CS-RBM continued to increase in quantity in the cyclosporin-treated group until the time of death (2-10 weeks post-transplantation). In the much longer lived combined transfusion and cyclosporin-treated groups, circulating antibody to BN-CS-RBM generally attained a maximum at approximately 2 to 4 months post-transplantation and then plateaued or decreased somewhat before the time of death (3-16 months post-transplantation). No correlation was found between quantity of circulating anti-BN-CS-RBM antibody and post-transplantation survival. Comparative study of the quantity of circulating antibody to BN-CS-RBM (the presumed nephritogenic antigen of experimental tubulointerstitial nephritis in the BN rat) in serum from transplant recipients as compared to serum from BN rats with severe experimental tubulointerstitial nephritis (TIN) (as induced by immunization with heterologous TBM antigens) demonstrated a greater quantity of potentially nephritogenic antibody circulating in transplant recipients than in BN rats with experimental TIN. Histologically, the transplanted kidneys in immunomodulated recipients demonstrated focal chronic interstitial inflammatory infiltrates with tubular atrophy and relative sparing of the glomeruli. The development of immune responses to tissue-specific alloantigens may become of clinical significance as graft-survival times are increased.


Purification and characterization of detergent stable alkaline protease from Bacillus amyloliquefaciens SP1 isolated from apple rhizosphere.
A thermostable extracellular alkaline protease producing Bacillus amyloliquefaciens SP1 was isolated from apple rhizosphere having multifarious plant growth promoting activities. Strain SP1 was purified to 6.48-fold using four-step purification protocol and characterized in detail for its robustness and ecofriendly application in leather and detergent industries. Structural analysis revealed that the protease was monomeric and had a molecular weight of 43 kDa. It exhibited optimum activity at 60°C in alkaline environment (pH 8.0) and stable in the presence of surfactants and oxidizing agents. Enzyme was thermostable at 50°C and retained more than 70% activity after 30 min incubation. It has shown stain removal property and dehairing of goat skin without chemical assistance and hydrolyzing fibrous proteins. This protease showed Km of 0.125 mg ml(-1) and V(max) of 12820 μg ml(-1) indicating its excellent affinity and catalytic role. Thermal inactivation of the pure enzyme followed first-order kinetics. The half life of the pure enzyme at 50, 60, and 65°C was 77, 19.80, and 13.33 min, respectively. The activation energy was 37.19 KJ mol(-1). The results suggest that the B. amyloliquefaciens SP1 has a potential application in different industries.


Treatment of Psoriasis With Biologic Therapy Is Associated With Improvement of Coronary Artery Plaque Lipid-Rich Necrotic Core: Results From a Prospective, Observational Study.
Lipid-rich necrotic core (LRNC), a high-risk coronary plaque feature assessed by coronary computed tomography angiography, is associated with increased risk of future cardiovascular events in patients with subclinical, nonobstructive coronary artery disease. Psoriasis is a chronic inflammatory condition that is associated with increased prevalence of high-risk coronary plaque and risk of cardiovascular events. This study characterized LRNC in psoriasis and how LRNC modulates in response to biologic therapy. Consecutive biologic naïve psoriasis patients (n=209) underwent coronary computed tomography angiography at baseline and 1-year to assess changes in LRNC using a novel histopathologically validated software (vascuCAP Elucid Bioimaging, Boston, MA) before and after biologic therapy over 1 year. Study participants were middle-aged, predominantly male with similar cardiometabolic and psoriasis status between treatment groups. In all participants at baseline, LRNC was associated with Framingham risk score (β [standardized β]=0.12 [95% CI, 0.00-0.15]; P=0.045), and psoriasis severity (β=0.13 [95% CI, 0.01-0.26]; P=0.029). At 1-year, participants receiving biologic therapy had a reduction in LRNC (mm2; 3.12 [1.99-4.66] versus 2.97 [1.84-4.35]; P=0.028), while those who did not receive biologic therapy over 1 year demonstrated no significant change with nominally higher LRNC (3.12 [1.82-4.60] versus 3.34 [2.04-4.74]; P=0.06). The change in LRNC was significant compared with that of the nonbiologic treated group (ΔLRNC, -0.22 mm2 versus 0.14 mm2, P=0.004) and remained significant after adjusting for cardiovascular risk factors and psoriasis severity (β=-0.09 [95% CI, -0.01 to -0.18]; P=0.033). LRNC was associated with psoriasis severity and cardiovascular risk factors in psoriasis. Additionally, there was favorable modification of LRNC in those on biologic therapy. This study provides evidence of potential reduction in LRNC with treatment of systemic inflammation. Larger, longer follow-up prospective studies should be conducted to understand how changes in LRNC may translate into a reduction in future cardiovascular events in psoriasis.


In vitro Epstein-Barr virus-immortalized lymphoma cell line carrying t(9;14)(p13;q32) chromosome abnormality, derived from splenic lymphoma with villous lymphocytes.
We herein describe splenic lymphoma with villous lymphocytes (SLVL) carrying t(9;14)(p13;q32). The t(9;14)(p13;q32) is a rare reciprocal chromosome translocation found in a subset of B-cell malignancies, mainly in low-grade non-Hodgkin's lymphomas. In t(9;14)(p13;q32), PAX-5 gene on 9p13 is involved with the immunoglobulin heavy-chain gene on 14q32. It has been thought that the deregulated expression of PAX-5 as a result of t(9;14)(p13;q32) may contribute to abnormal cell proliferation. Although continuous cell lines are invaluable tools for studying lymphomagenesis in the t(9;14)(p13;q32)-bearing lymphomas, establishment of such cell lines is extremely difficult since they are usually mature B-cell malignancies. In an attempt to transform the SLVL cells into a proliferating cell line, we examined the responses of the cells to infection by Epstein-Barr virus (EBV). SLVL cells were found to be susceptible to immortalization by EBV, resulting in a permanent cell line. The cell line, designated SL-15, possessed the t(9;14)(p13;q32). Genotype analysis and immunophenotype profiles confirmed that the cell line arose from the primary lymphoma cells. The cells had characteristic cytoplasmic villi. SL-15 cells has been growing over 2 years equivalent to 350-400 population doubling levels without proliferative crisis that is often observed in EBV-positive lymphoblastoid cell lines. Furthermore, SL-15 cells, when inoculated into nude mice, formed t(9;14)(p13;q32)-bearing tumors with cytoplasmic villi. The validated SLVL-derived cell line provide a useful model system to study molecular biology of t(9;14)(p13;q32)-bearing B-cell malignancies as well as lymphomagenesis of SLVL in vitro and in vivo.


A random forest based biomarker discovery and power analysis framework for diagnostics research.
Biomarker identification is one of the major and important goal of functional genomics and translational medicine studies. Large scale -omics data are increasingly being accumulated and can provide vital means for the identification of biomarkers for the early diagnosis of complex disease and/or for advanced patient/diseases stratification. These tasks are clearly interlinked, and it is essential that an unbiased and stable methodology is applied in order to address them. Although, recently, many, primarily machine learning based, biomarker identification approaches have been developed, the exploration of potential associations between biomarker identification and the design of future experiments remains a challenge. In this study, using both simulated and published experimentally derived datasets, we assessed the performance of several state-of-the-art Random Forest (RF) based decision approaches, namely the Boruta method, the permutation based feature selection without correction method, the permutation based feature selection with correction method, and the backward elimination based feature selection method. Moreover, we conducted a power analysis to estimate the number of samples required for potential future studies. We present a number of different RF based stable feature selection methods and compare their performances using simulated, as well as published, experimentally derived, datasets. Across all of the scenarios considered, we found the Boruta method to be the most stable methodology, whilst the Permutation (Raw) approach offered the largest number of relevant features, when allowed to stabilise over a number of iterations. Finally, we developed and made available a web interface ( https://joelarkman.shinyapps.io/PowerTools/ ) to streamline power calculations thereby aiding the design of potential future studies within a translational medicine context. We developed a RF-based biomarker discovery framework and provide a web interface for our framework, termed PowerTools, that caters the design of appropriate and cost-effective subsequent future omics study.


Identification of a novel in-frame de novo mutation in SPTAN1 in intellectual disability and pontocerebellar atrophy.
Heterozygous in-frame mutations (p.E2207del and p.R2308_M2309dup) in the α-II subunit of spectrin (SPTAN1) were recently identified in two patients with intellectual disability (ID), infantile spasms (IS), hypomyelination, and brain atrophy. These mutations affected the C-terminal domain of the protein, which contains the nucleation site of the α/β spectrin heterodimer. By screening SPTAN1 in 95 patients with idiopathic ID, we found a de novo in-frame mutation (p.Q2202del) in the same C-terminal domain in a patient with mild generalized epilepsy and pontocerebellar atrophy, but without IS, hypomyelination, or other brain structural defects, allowing us to define the core phenotype associated with these C-terminal SPTAN1 mutations. We also found a de novo missense variant (p.R566P) of unclear clinical significance in a patient with non-syndromic ID. These two mutations induced different patterns of aggregation between spectrin subunits in transfected neuronal cell lines, providing a paradigm for the classification of candidate variants.


Colorimetric dimethyl sulfide sensor using Rhodovulum sulfidophilum cells based on intrinsic pigment conversion by CrtA.
A colorimetric whole-cell sensor for dimethyl sulfide (DMS) was constructed based on the in vivo conversion of intrinsic pigments in response to the analyte. In a marine bacterium, Rhodovulum sulfidophilum, carotenoids are synthesized via the spheroidene pathway. In this pathway, demethylspheroidene, a yellow carotenoid, is converted to spheroidene under catalysis of O-methyltransferase. Spheroidene monooxygenase (CrtA) catalyzes the terminal step of the pathway and converts spheroidene to spheroidenone, a red carotenoid. Here, the CrtA gene in R. sulfidophilum was removed and then reintroduced downstream of the DMS dehydrogenase gene promoter. Using this whole-cell sensor, 3 muM DMS or dimethyl sulfoxide can be detected without adding any color-forming reagent. The ratio of the red spheroidenone to total carotenoids increased, as the DMS concentration was raised to 0.3 mM. Comparison of the signal to the background color indicated a shift in the color coordinate from a yellow to a red hue. An intense signal was obtained with 1-day incubation at a high cell density when sensor cells at the exponential growth phase were used. These results show that the genetically engineered R. sulfidophilum cells can be used to monitor the quality of marine aquacultural environments by the naked eye.


Lipid surface charge does not influence conductance or calcium block of single sodium channels in planar bilayers.
We have studied the effects of membrane surface charge on Na+ ion permeation and Ca2+ block in single, batrachotoxin-activated Na channels from rat brain, incorporated into planar lipid bilayers. In phospholipid membranes with no net charge (phosphatidylethanolamine, PE), at low divalent cation concentrations (approximately 100 microM Mg2+), the single channel current-voltage relation was linear and the single channel conductance saturated with increasing [Na+] and ionic strength, reaching a maximum (gamma max) of 31.8 pS, with an apparent dissociation constant (K0.5) of 40.5 mM. The data could be approximated by a rectangular hyperbola. In negatively charged bilayers (70% phosphatidylserine, PS; 30% PE) slightly larger conductances were observed at each concentration, but the hyperbolic form of the conductance-concentration relation was retained (gamma max = 32.9 pS and K0.5 = 31.5 mM) without any preferential increase in conductance at lower ionic strengths. Symmetrical application of Ca2+ caused a voltage-dependent block of the single channel current, with the block being greater at negative potentials. For any given voltage and [Na+] this block was identical in neutral and negatively charged membranes. These observations suggest that both the conduction pathway and the site(s) of Ca2+ block of the rat brain Na channel protein are electrostatically isolated from the negatively charged headgroups on the membrane lipids.


Risk factors for complications and return to the emergency department after interscalene block using liposomal bupivacaine for shoulder surgery.
Exparel (liposomal bupivacaine) has recently gained favor for use in interscalene regional blocks for shoulder surgery. While effective for pain relief, this does have adverse effects that can lead to postoperative emergency department (ED) visits. This study aimed to identify any patient risk factors that are associated with complications leading to ED return visits owing to interscalene blocks using Exparel before shoulder surgery. A retrospective chart review was performed for all patients undergoing shoulder surgery with an Exparel interscalene block in an 8-month period. For each patient, demographic information, comorbidities, type of block, postoperative complications, ED return visits, and readmissions were recorded. The 5-factor modified Frailty Index score and the Charlson Comorbidity Index score were calculated. Univariate and multivariate logistic regressions were conducted to identify risk factors associated with increased complications and return to the ED. Overall, 352 patients were included; most patients were men, were aged between 51 and 70 years, and had a body mass index of 25.0-35.0. Postoperative complications related to the Exparel interscalene block occurred in 58 patients (16.5%), including 37 minor complications (10.5%) and 21 major complications (6.0%) that led to return ED visits. Univariate analysis yielded American Society of Anesthesiologists (ASA) score (P = .03) as a significant predictor of minor complications. Multivariate logistic regression analysis yielded ASA score (P = .096; odds ratio, 1.64) as trending toward being a significant risk factor for minor complications. Univariate analysis yielded age (P = .006), ASA score (P = .009), and Charlson Comorbidity Index score (P = .002) as significant predictors of major complications. Multivariate logistic regression analysis yielded ASA score (P = .049; odds ratio, 2.25) as the only significant risk factor for major complications. Surgeons and anesthesiologists should strongly consider a patient's ASA score, in addition to his or her pulmonary and cardiac history, when deciding whether the patient is an appropriate candidate for an interscalene regional block using Exparel for shoulder surgery.


The effects of small but abrupt change in temperature on the behavior of larval zebrafish.
The zebrafish has become increasingly utilized in behavioral neuroscience, psychopharmacology, and behavior genetics research. However, little attention has been paid to the potential effects of environmental conditions fish are exposed to before and during behavioral testing. One such important factor is temperature, more specifically, the difference in temperature between holding and test tanks. In the current study, we bred and raised zebrafish of the AB strain in 28 °C water for 7 days until they were free swimming. On the 7th day we placed each experimental subject singly into the testing well with water set to 24 °C, 28 °C or 30 °C for 15 min and recorded the behavior of the subjects. We found significant and behavior-specific idiosyncratic effects of the employed temperature changes. For example, lowering the temperature decreased speed, but increasing the temperature did not alter it. Increased temperature reduced duration of immobility and reduced absolute turn angle, but lowered temperature did not alter these behaviors. While lowered temperature, and to a lesser degree also increased temperature, reduced intra-individual temporal variance of absolute turn angle. Furthermore, we found no change in thigmotaxis and frequency of immobility by either temperature change. Our results demonstrate the importance of temperature in behavioral studies with zebrafish and suggest that equating water temperature between holding and testing tanks is required to enhance reproducibility and replicability of results with this species.


Thermoregulatory and maternal nestbuilding in Syrian hamsters: interaction of ovarian steroids and energy demand.
To investigate the relationship between energy demand, ovarian hormones and behavioral thermoregulation, we measured nestbuilding of female Syrian hamsters (Mesocricetus auratus) under conditions where energy demand and circulating ovarian hormone levels varied. Hamsters increased nestbuilding under conditions of chronically elevated estradiol and progesterone levels (late gestation; ovariectomized animals treated with estradiol +/- progesterone) and increased energy demand (late gestation or cold exposure). When hormone treatments were withdrawn, hamsters gained weight and reduced nestbuilding activity. Nestbuilding was not affected by lactation (characterized by low ovarian steroid levels and high energy demand) or by estrous cycle phase (characterized by relatively transient changes in ovarian steroid levels and subtle changes in energy demand). Thus, nestbuilding in Syrian hamsters was responsive to increased energy demand from cold exposure, late gestation, and to ovarian hormones. However, during lactation and estrous cycles, nestbuilding did not always change predictably according to known changes in energy expenditure and steroid hormone levels. Thus, levels of nestbuilding are influenced by factors in addition to ovarian hormones and energy expenditure under some circumstances.


Comparing left atrial indices by CMR in association with left ventricular diastolic dysfunction and adverse clinical outcomes.
Left atrial (LA) features are altered when diastolic dysfunction (DD) is present. The relations of LA features to the DD severity and to adverse outcomes remain unclear using CMR images. We sought to compare LA features including volumes, emptying fraction, and strains as predictors of left ventricular (LV) DD and adverse outcomes. We compared four groups including normal controls (n = 32), grade I DD (n = 69), grade II DD (n = 42), and grade III DD (n = 21). DD was graded by echocardiography following the current ASE guidelines. Maximum LA volume (LAVmax), minimum LA volume (LAVmin), and LA emptying fraction (LAEF) were assessed using CMR cine images. Phasic LA strains including reservoir, conduit, and booster pump strain were assessed by feature tracking. The outcome was a composite of hospital admissions for heart failure and all-cause mortality analyzed using Cox proportional hazard models. LAVmax and LAVmin were progressively larger while LAEF and LA strain measures were lower with worsening degree of DD (all p < 0.001). Among 132 patients with DD, 61 reached the composite outcome after on average 36-months of follow-up. Each of the LA parameters except for LA conduit strain was an independent predictor of the outcome in the adjusted Cox proportional hazard models (all p < 0.001). They remained significant outcome predictors after the model additionally adjusted for LV longitudinal strain. The AUC of outcome prediction was highest by LAEF (0.760) followed by LA reservoir strain (0.733) and LAVmin (0.725). Among all the LA features, increased LA volumes, reduced LAEF, reduced LA reservoir and booster pump strains were all associated with DD and DD severity. While LA strains are valuable, conventional parameters such as LAEF and LAVmin remain to be highly effective in outcome prediction with comparable performance.


Acute Poisoning with Dapsone and Olanzapine: Severe Methemoglobinemia and Coma with a Favourable Outcome.
Dapsone is a drug commonly used in the treatment of leprosy. In Europe it is rarely prescribed, mostly for the treatment of skin diseases such as dermatitis herpetiformis. Poisoning with dapsone is rare and reports of such cases are of interest for toxicological practice. We describe the only acute dapsone poisoning in a caseload series of 21,000 intoxications treated in the Clinical Toxicology Clinic at St George University Hospital in Plovdiv, Bulgaria between 1999 and 2013. We report on a 36-year-old woman who attempted deliberate self-poisoning with an ingestion of approximately 4.5 g of dapsone and 0.3 g of olanzapine. On admission, the patient was in a state of severe intoxication and comatose. On admission to hospital 9 hours after the ingestion, the methemoglobin level was 51.7%. The patient recovered 8 days later. She received complex treatment including intubation, ventilation, repeated gastric lavage, hemodialysis, blood exchange transfusion and antidote treatment with methylene blue. She was discharged in good clinical condition with minimal organ damage such as mild toxic hepatitis.


Circularly Polarized Luminescence of a Stereogenic Curved Paraphenylene Anchoring a Chiral Binaphthyl in Solution and Solid State.
Invited for the cover of this issue are Masashi Hasegawa, Yoshitane Imai, Yasuhiro Mazaki and co-workers at Kitasato University, Kindai University and Osaka University. The image depicts a stereogenic twisted cycloparaphenylene that exhibits circularly polarized luminescence in a polymethyl methacrylate thin film. Read the full text of the article at 10.1002/chem.202004283.


The diapause-like colorectal cancer cells induced by SMC4 attenuation are characterized by low proliferation and chemotherapy insensitivity.
In response to adverse environmental conditions, embryonic development may reversibly cease, a process termed diapause. Recent reports connect this phenomenon with the non-genetic responses of tumors to chemotherapy, but the mechanisms involved are poorly understood. Here, we establish a multifarious role for SMC4 in the switching of colorectal cancer cells to a diapause-like state. SMC4 attenuation promotes the expression of three investment phase glycolysis enzymes increasing lactate production while also suppressing PGAM1. Resultant high lactate levels increase ABC transporter expression via histone lactylation, rendering tumor cells insensitive to chemotherapy. SMC4 acts as co-activator of PGAM1 transcription, and the coordinate loss of SMC4 and PGAM1 affects F-actin assembly, inducing cytokinesis failure and polyploidy, thereby inhibiting cell proliferation. These insights into the mechanisms underlying non-genetic chemotherapy resistance may have significant implications for the field, advancing our understanding of aerobic glycolysis functions in tumor and potentially informing future therapeutic strategies.


A novel plant glutathione peroxidase-like protein provides tolerance to oxygen radicals generated by paraquat in Escherichia coli.
Citrus salt-stress associated protein (Cit-SAP) reveals significant sequence homology to mammalian glutathione peroxidase (GP). In an attempt to assign biological function to this protein, transformed E. coli cells expressing Cit-SAP were examined for their ability to tolerate free radicals formed by paraquat, an O2- radical forming agent. In the presence of paraquat, the survival rate of the transformed bacteria expressing Cit-SAP was much higher as compared to the wild-type bacteria. The results support the assumption that Cit-SAP is a plant GP-like protein which participate in the enzymatic system aimed at scavenging oxygen free-radicals in plants.


Filter tilting and retrievability of the Celect and Denali inferior vena cava filters using propensity score-matching analysis.
To assess the filter tilting and outcomes of the Celect and Denali inferior vena cava (IVC) filters by using a propensity score-matching analysis. From January 2009 to November 2017, 181 Celect and 58 Denali filters were inserted in our institution. To assess filter tilt, filter tip abutment or penetration of the IVC wall, and retrieval outcome, independent variables, including age, sex, IVC long diameter, IVC angulation, and proximity of the filter to renal vein insertion, were entered in the propensity model. Comparative analyses were performed before and after propensity score-matching analysis. Thirty-one patients were enrolled in each group for the final propensity score-matching analysis. The mean filter indwelling time was not significantly different between the groups (26 ± 22 days in Celect and 27 ± 23 days in Denali). After propensity score adjustment, the mean degree of filter tilt was higher in the Celect group (9.5° ± 7.4° vs 5.6° ± 6.7°). Filter tip abutment or penetration of the IVC wall was more common in the Celect group (39% [12/31, abutment: 12, penetration: 0] vs 13% [4/31, abutment: 3, penetration: 1]). The retrieval outcomes were not significantly different before and after propensity score adjustment in both filters. The Denali IVC filter showed less tilt and low rate of filter tip abutment to the IVC wall after propensity score-matching analysis. The retrieval rate was not significantly different in the short-term filter indwelling setting. More large-scale, long-term follow-up studies are needed to verify these results.


Hierarchical modeling of blood pressure determinants and outcomes following valsartan treatment in hypertensive patients with known comorbidities: pooled analysis of six prospective real-world studies including 11,999 patients.
Six prospective real-world studies of antihypertensive treatment with valsartan-centric regimens were pooled to: (1) examine the effectiveness of ∼90 days of second- or later-line valsartan treatment in hypertensive patients with known comorbidities; and (2) identify physician- and patient-related determinants associated with systolic (SBP) and diastolic blood pressure (DBP) outcomes in these patients. A pooled analysis was performed of an evaluable sample of 11,999 hypertensive patients with known comorbidities treated ∼90 days with valsartan-centric regimens. We applied hierarchical linear and logistic regression models to identify determinants of blood pressure (BP) outcomes and a potential physician class effect. Valsartan regimens resulted in mean (SD) SBP and DBP reductions of 18.0 (15.8) mmHg and 9.5 (10.1) mmHg, respectively, at ∼90 days, yielding SBP, DBP and combined SBP/DBP control rates of 44.0%, 67.2% and 39.3%, respectively. About a quarter of the variance in 90 day BP values was attributable to a physician class effect. BP outcomes declined with physicians' increasing years in practice and being male. At the patient level, BP outcomes declined with SBP and DBP at diagnosis; diabetes; higher cholesterol and BMI; lower valsartan and hydrochlorothiazide (HCTZ) doses; and concomitant anti-hypertensives. Older age was associated with improved DBP. A proxy of physician vigilance, cardiovascular disease history, was associated with improved BP outcomes, as were patient adherence and higher doses of valsartan in combination with HCTZ. Valsartan-centric regimens have significant BP lowering benefits in this pooled sample of patients with known comorbidities. Many observed determinants of BP outcomes are modifiable or manageable.


Lung eosinophils increase vagus nerve-mediated airway reflex bronchoconstriction in mice.
Eosinophils mediate airway hyperresponsiveness by increasing vagally mediated reflex bronchoconstriction. Here, we tested whether circulating or airway eosinophils change nerve function. Airway resistance in response to aerosolized 5-hydroxytryptamine (5-HT, 10-300 mM) was measured in wild-type mice or transgenic mice that overexpress IL5 in T cells (+IL5T), overexpress IL5 in airway epithelium (+IL5AE), or overexpress IL5 but are devoid of eosinophils (+IL5AE/-Eos). Inflammatory cells in bronchoalveolar lavage (BAL), blood, and bone marrow were quantified. Blood eosinophils were increased in +IL5T and +IL5AE mice compared with wild-type mice. +IL5T mice had increased eosinophils in bone marrow while +IL5AE mice had increased eosinophils in BAL. Eosinophils surrounding large airways were significantly increased only in +IL5AE mice. With intact vagal innervation, aerosolized 5-HT significantly increased airway resistance in +IL5AE mice. 5-HT-induced bronchoconstriction was blocked by vagotomy or atropine, demonstrating that it was mediated via a vagal reflex. Airway resistance was not increased in +IL5AE/-Eos mice, demonstrating that it required lung eosinophils, but was not affected by increased bone marrow or blood eosinophils or by increased IL5 in the absence of eosinophils. Eosinophils did not change M3 function on airway smooth muscle, since airway responses to methacholine in vagotomized mice were not different among strains. Eosinophils surrounding large airways were sufficient, even in the absence of increased IL5 or external insult, to increase vagally mediated reflex bronchoconstriction. Specifically blocking or reducing eosinophils surrounding large airways may effectively inhibit reflex hyperresponsiveness mediated by vagus nerves in eosinophilic asthma.


Domain Architecture Based Methods for Comparative Functional Genomics Toward Therapeutic Drug Target Discovery.
Genes duplicate, mutate, recombine, fuse or fission to produce new genes, or when genes are formed from de novo, novel functions arise during evolution. Researchers have tried to quantify the causes of these molecular diversification processes to know how these genes increase molecular complexity over a period of time, for instance protein domain organization. In contrast to global sequence similarity, protein domain architectures can capture key structural and functional characteristics, making them better proxies for describing functional equivalence. In Prokaryotes and eukaryotes it has proven that, domain designs are retained over significant evolutionary distances. Protein domain architectures are now being utilized to categorize and distinguish evolutionarily related proteins and find homologs among species that are evolutionarily distant from one another. Additionally, structural information stored in domain structures has accelerated homology identification and sequence search methods. Tools for functional protein annotation have been developed to discover, protein domain content, domain order, domain recurrence, and domain position as all these contribute to the prediction of protein functional accuracy. In this review, an attempt is made to summarise facts and speculations regarding the use of protein domain architecture and modularity to identify possible therapeutic targets among cellular activities based on the understanding their linked biological processes.


Infliximab counteracts tumor necrosis factor-α-enhanced induction of matrix metalloproteinases that degrade claudin and occludin in non-pigmented ciliary epithelium.
Infliximab, a monoclonal antibody directed against human tumor necrosis factor-alpha (TNF-α), effectively treats anterior uveitis, which can accompany Behçet's disease. Here, we investigated the underlying mechanism of this action. We examined human, non-pigmented ciliary epithelial cells (HNPCECs), which make up the blood-aqueous barrier (BAB) in the uvea. We measured the expression levels of matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs in the presence or absence of TNF-α using quantitative, real-time polymerase chain reaction and enzyme-linked immunosorbent assays. The expression of MMP-1, MMP-3, and MMP-9 increased in the presence of TNF-α, and the addition of infliximab reversed the increase. The TNF-α effects were more attenuated when infliximab was added before than when it was added after TNF-α exposure. Gelatin zymography demonstrated that the protease activity of these MMPs was also increased in the presence of TNF-α and attenuated with infliximab. Immunostaining showed that MMP-1, MMP-3, and MMP-9 degraded claudin-1 and occludin in HNPCECs and in non-pigmented ciliary epithelial cells of the swine ciliary body. In a monolayer of HNPCECs, we found that permeability was significantly increased with MMP treatment. Thus, TNF-α increased levels of MMPs in cells that form the BAB, and MMPs degraded components of the tight junctions in the BAB, which increased permeability through the cellular barrier. Furthermore, infliximab effectively attenuated the TNF-α-induced increases in MMP expression in cells that make up the BAB. These findings might suggest a basis for the clinical prevention of anterior uveitis.


Temporal cohesion as a candidate for negentropy in biological thermodynamics.
Negentropy explored by Schrödinger for saving biology within thermodynamics stands firm on the perpetual temporal cohesion of material origin. The temporal cohesion refers to the cohesion acting between what has been produced and what is going to produce, thus keeping the negentropy as a measure of organization constantly positive in time. Such cohesion is ubiquitous in measurement internal to the material world. Internal measurement in the quantum realm really lets the current "detecting" constantly feed upon some quantum resources available from the preceding "detected" a moment ago. The quantum resources transferred during the cohesive process serve as a physical factor for bridging between the two different temporalities; the present perfect and progressive tenses. What has been detected comes to participate in an attribute of what is going to detect next as constantly following suit. Temporal cohesion is an agential mediator connecting the adjacent temporalities in a manner different from the spatial cohesion observing only the single temporality restricted to the present tense. The activity of enhancing the magnitude of negentropy may have preceded the emergence of something called life phenomenon. Biology feeds upon temporal cohesion.


Portable current stimulator for transdermal iontophoretic drug delivery.
A pulse stimulator was designed. It is small sized (3 x 2 cm printed circuit board) and battery-powered (185 microA total static current). The current intensity and pulse duration of this device can be continuously varied. Preliminary trials of lidocaine show that this device is usable for transdermal drug delivery and may be valuable for portable applications.


Dopamine D2 receptor modulates Wnt expression and control of cell proliferation.
The Wnt/β-catenin pathway is one of the most conserved signaling pathways across species with essential roles in development, cell proliferation, and disease. Wnt signaling occurs at the protein level and via β-catenin-mediated transcription of target genes. However, little is known about the underlying mechanisms regulating the expression of the key Wnt ligand Wnt3a or the modulation of its activity. Here, we provide evidence that there is significant cross-talk between the dopamine D2 receptor (D2R) and Wnt/β-catenin signaling pathways. Our data suggest that D2R-dependent cross-talk modulates Wnt3a expression via an evolutionarily-conserved TCF/LEF site within the WNT3A promoter. Moreover, D2R signaling also modulates cell proliferation and modifies the pathology in a renal ischemia/reperfusion-injury disease model, via its effects on Wnt/β-catenin signaling. Together, our results suggest that D2R is a transcriptional modulator of Wnt/β-catenin signal transduction with broad implications for health and development of new therapeutics.


Physician code creep: evidence in Medicaid and State Employee Health Insurance billing.
This study estimates a fixed effects ordered logit model physician office visit billing using claims data from South Carolina Medicaid and the State Employees Health Plan. The results find code creep increasing expenditures on physician office visits at a rate of 2.2 percent annually for both programs, with no significant difference in the rate between the two. The models also indicate that physician billing patterns differ between the programs, with the Medicaid claims averaging 1.3 percent less per visit than comparable State Employees Health Plan claims.


Restriction of range effects in post-metabolic and bariatric surgery outcomes research: considerations for clinical decision making.
Preoperative psychopathology does not consistently predict postoperative outcomes in patients who undergo metabolic and bariatric surgery (MBS). Individuals with elevated pre-MBS psychopathology may be less likely to undergo surgery, which may create a floor effect given the limited range of scores on measures of psychopathology included in postoperative analyses, thereby decreasing the power to detect clinically significant differences between groups. Our objective was to compare rates of clinically significant pre-MBS psychopathology across domains of functioning in patients who did and did not undergo MBS: surgical completers (SCs, n = 286) and nonsurgical completers (NSCs, n = 125). Academic medical center, United States. Participants (n = 411) were a racially diverse sample of MBS candidates who completed a preoperative psychosocial evaluation including measures of disordered eating, alcohol and tobacco use, pain catastrophizing, anxiety, and depressive symptomatology. Compared with SCs, NSCs had larger scale score variance on measures of psychopathology and were more likely to be Black; to report clinically significant scores on measures of binge eating, depression, anxiety, and pain catastrophizing; and to use tobacco. Results support the presence of a restriction-of-range effect but do not demonstrate a floor effect. These data suggest that current outcome data for MBS patients may not generalize to those who report clinically significant psychopathology at the pre-MBS psychosocial evaluation and may warrant caution when using the current literature to inform clinical decision making for this group. Findings also suggest a need for interventions that will better engage Black patients.


Bias correction of regional climate model simulations for the impact assessment of the climate change in Egypt.
This study projected the future temperature change for Egypt during the late of this century (2071-2100) for three representative concentration pathways (RCP2.6, RCP4.5, and RCP8.5), by correcting regional climate model (RCM) simulations of average, maximum, and minimum daily temperature with reference to observed data of 26 stations. Four commonly used methods of bias correction have been applied and evaluated: linear scaling, variance scaling, and theoretical and empirical quantile mapping. The compromise programing results of the applied evaluation criteria show that the best method is the variance scaling, and thus it was applied to transfer the correction factor to the projections. All temperature indices are expected to increase significantly under all scenarios and reach the highest record by the end of the century, i.e., the expected increase in average, maximum, and minimum temperature ranges between 4.08-7.41 °C, 4.55-7.89 °C, and 3.88-7.23 °C, respectively. The largest temperature rise will occur in the summer, with the highest increase in the maximum (minimum) temperature of 10.9 °C (10 °C) in July and August under RCP8.5. The maximum (minimum) winter temperature, on the other hand, will drop by a maximum of 2 °C (1.35 °C) under RCP2.6. The Western Desert and Upper Egypt are the regions most affected by climate change, while the northern region of Egypt is the least affected. These findings would help in impact assessment and adaptation strategies and encourage further investigation to evaluate various climate models in order to obtain a comprehensive assessment of the climate change impacts on different hydrometeorological processes in Egypt.


Changes in the convulsion threshold and in the level of brain biogenic amines in rats chronically treated with phenobarbital or diazepam.
In experiments on 72 male Wistar rats, chronically treated with phenobarbital and diazepam, studies are made of the change in the convulsion threshold after sound stimulation during treatment and after withdrawal of the drugs. The level of biogenic amines in the brain at the end of the treatment (60th day) and after withdrawal of the drugs (3rd day) was also determined. The convulsion threshold was found to be considerably lower on the 3rd day after drug withdrawal in rats treated with phenobarbital and to a lesser extent (also on the 3rd day) in rats treated with diazepam. Serotonin level is considerably lower in both groups of animals. The significance of the disturbed correlation between serotonin and noradrenaline and particularly the sharp decrease of the serotonin level for the appearance of intensified convulsion activity is discussed.


Acute Electrical Pulse Stimulation and Hyperglycemia Regulates RCAN1-4 in C2C12 myotubes through Oxidative Stress.
Regulator of Calcineurin1 (RCAN1) controls the Sr/Thr phosphatase, Calcineurin. RCAN1 has never been studied in-vitro in myotubes following acute Electrical Pulse Stimulation (EPS) which recapitulates the physiological effect of exhaustive exercise and hyperglycemia (HG). 144 h differentiated C2C12 myotubes were treated either with high glucose (HG) at 15mM or kept as control for 48 h. The myotubes were subjected to EPS for 1second followed by 1second of pause for 90min. Two separate Non-EPS controls with and without HG were performed simultaneously. The protein level of RCAN1-4 increased immediately after EPS up to 6hours in HG myotubes and in control. Non-EPS myotubes treated with HG exhibited a higher level of RCAN1-4 compare to Non-EPS control. RCAN1-1L and 1-S did not show any significant change after EPS in both groups. Calcineurin level decreased immediately after EPS. The levels of CuZn SOD and MnSOD were increased immediately after EPS whereas Catalase level increased significantly only 3hours after EPS in both groups. CuZn SOD and MnSOD showed higher level in HG control compare to control while the level of Catalase was lesser in HG control. NFATc4 level decreased at 3Hr post EPS in both groups. PGC1-? remained unchanged in all the samples. 4HNE increased significantly in both groups after EPS and was higher in HG control than in control. Protein Carbonyl (PC) increased significantly 3hours after EPS in normal cells and returned to basal level at 6 h. However in HG myotubes, PC level increased immediately after EPS and no further modification until 6Hr was observed. HG samples exhibited much higher PC than normal samples. We have already shown than exhaustive exercise regulates RCAN1-4 in rat skeletal muscle through involvement of oxidative stress. The acute EPS of C2C12 myotubes in normal condition or under hyperglycemia, served as an in-vitro model of exercise and diabetic regulation of RCAN1 through oxidative stress in-vitro.


Tailoring Hydrogel Composition and Stiffness to Control Smooth Muscle Cell Differentiation in Bioprinted Constructs.
Reliable in vitro cellular models are needed to study the phenotypic modulation of smooth muscle cells (SMCs) in health and disease. The aim of this study was to optimize gelatin methacrylate (GelMA)/alginate hydrogels for bioprinting three-dimensional (3D) SMC constructs. Four different hydrogel groups were prepared by mixing different concentrations (% w/v) of GelMA and alginate: G1 (5/1.5), G2 (5/3), G3 (7.5/1.5), and G4 (7.5/3). GelMA 10% was used as control (G5). A circular structure containing human bladder SMCs was fabricated by using an extrusion-based bioprinter. The effects of the mixing ratios on printability, viability, proliferation, and differentiation of the cells were investigated. Rheological analysis showed that the addition of alginate significantly stabilized the change in mechanical properties with temperature variations. The group with the highest GelMA and alginate concentrations (G4) exhibited the highest viscosity, resulting in better stability of the 3D construct after crosslinking. Compared to other hydrogel compositions, cells in G4 maintained high viability (> 80%), exhibited spindle-shaped morphology, and showed a significantly higher proliferation rate within an 8-day period. More importantly, G4 provided an optimal environment for the induction of a SMC contractile phenotype, as evidenced by significant changes in the expression of marker proteins and morphological parameters. Adjusting the composition of GelMA/alginate hydrogels is an effective means of controlling the SMC phenotype. These hydrogels support bioprinting of 3D models to study phenotypic smooth muscle adaptation, with the prospect of using the constructs in the study of therapies for the treatment of urethral strictures.


Effects of taraxasterol on inflammatory responses in lipopolysaccharide-induced RAW 264.7 macrophages.
Taraxasterol, a pentacyclic-triterpene, was isolated from the Chinese medicinal herb Taraxacum officinale. In the present study, we investigated the in vitro anti-inflammatory activity of taraxasterol in lipopolysaccharide (LPS)-induced RAW 264.7 murine macrophages. RAW 264.7 cells were pretreated with 2.5, 5, or 12.5μg/ml of taraxasterol 1h prior to treatment with 1μg/ml of LPS. Nitric oxide (NO) level in supernatants from cells was examined by Griess reaction, the concentrations of prostaglandin E(2) (PGE(2)), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) were measured by ELISA. Nuclear factor kappa B (NF-κB) activation was evaluated by immunocytochemical analysis. We found that taraxasterol inhibited NO, PGE(2), TNF-α, IL-1β and IL-6 production in LPS-induced RAW 264.7 macrophages in a dose-dependent manner. Further studies revealed that taraxasterol prevented the LPS-induced NF-κB translocation from cytoplasm into nuclear. These results indicate that taraxasterol has anti-inflammatory effect by blocking NF-κB pathway.


A randomized controlled trial of amantadine plus interferon-alpha2a vs. interferon-alpha2a alone in naive patients with chronic hepatitis C randomized according to the early virological response to interferon-alpha2a monotherapy.
An early virological response to interferon-alpha treatment is a strong predictor of sustained response, but it has never been exploited to stratify patients in clinical trials. To evaluate the efficacy of amantadine plus interferon-alpha compared with interferon-alpha alone in naive patients with chronic hepatitis C who were randomized on the basis of the early virological response to interferon-alpha. One hundred and eighty-one patients received recombinant interferon-alpha2a (3 MU three times weekly) for 2 months and 164 were evaluated for early (i.e. month 2) virological response. Hepatitis C virus (HCV) RNA-negative patients (n = 66) were randomized to receive 3 MU of interferon-alpha three times weekly, with or without amantadine (200 mg/day); HCV RNA-positive patients (n = 98) were randomized to receive 6 MU of interferon-alpha three times weekly, with or without amantadine (200 mg/day). HCV RNA-positive patients at 6 months discontinued treatment, and all others completed 12 months. At month 6, HCV RNA-negative patients made up 54.2% of the interferon + amantadine group and 42.0% of the monotherapy group (P = 0.07). At month 12, HCV RNA-negative patients made up 38.5% of the interferon + amantadine group and 28.4% of the monotherapy group (N.S.). The sustained virological response rates were 21.6% and 20.9%, respectively (N.S.). The addition of amantadine does not enhance the sustained virological response to interferon-alpha in naive patients with chronic hepatitis C; however, an additive effect of amantadine occurs in the first 6 months, mainly in patients without an early response to monotherapy. Early response to interferon-alpha is a strong predictor of sustained virological response.