Development and Validation of Predictive Assessment of Complicated Diverticulitis Score.
The prevalence of acute diverticulitis (AD) has progressively increased in recent decades, with correspondingly greater morbidity and mortality. The aim of the study is to develop a predictive score to identify patients with the highest risk of complicated AD. The clinical records of 1089 patients referred to the emergency department (ED) over a five-year period were reviewed. In multivariate analysis, male sex (p < 0.001), constipation (p = 0.002), hemoglobin < 11.9 g/dL (p < 0.001), C reactive protein > 80 mg/L (p < 0.001), severe obesity (p = 0.049), and no proton pump inhibitor treatment (p = 0.003) were independently associated with complicated AD. The predictive assessment of complicated (PACO)-diverticulitis (D) score, including these six variables, was applied to the retrospective cohort and then validated prospectively in a cohort including 282 patients. It categorized patients into three risk classes for complicated AD. The PACO-D score showed fair discrimination for complicated AD with an area under the receiver operating characteristic curve of 0.674 and 0.648, in the retrospective and prospective cohorts, respectively. The PACO-D score could be a practical clinical tool to identify patients at highest risk for complicated AD referred to the ED so that appropriate diagnostic and therapeutic resources could be appropriately allocated. Further external validation is needed to confirm these results.


Vision improvement in severe acute isolated optic neuritis after plasma exchange treatment in Chinese population: a prospective case series study.
Plasma exchange (PE) is often considered as an effective treatment for neuromyelitis optica spectrum disorder (NMOSD) and several inflammatory demyelinating disorders of the central nervous system. This study aimed to evaluate the visual outcomes of Chinese patients with severe acute isolated optic neuritis (ON) who received PE therapy after high-dose intravenous methylprednisolone (IVMP) treatment. Thirty-seven acute isolated ON patients experiencing their first attack of severe visual impairment without neurological disability were recruited. All subjects received five cycles of double-filtration plasmapheresis. Visual acuity (VA) was documented at onset, 1 day before PE treatment, at each cycle of PE therapy and at the 1-month follow-up visit. This study included 26 female (70.3%) and 11 male (29.7%) subjects, and 18 subjects (48.6%) had bilateral involvement. The time window between onset and PE treatment was 27.3 ± 12.7 days (range: 6-53 days). Mean VA (logMAR) of the studied eyes at onset, 1-day before PE treatment/after IVMP and after the fifth PE treatment were 3.41 ± 1.50, 2.61 ± 1.64 and 1.66 ± 1.52, respectively (p < 0.001). Nineteen eyes (51.4%) showed no light perception at the onset, and 17 eyes (45.9%) improved to Snellen VA >20/800 after IVMP and PE treatments, among which five eyes (13.5%) recovered to Snellen VA 20/20 (p < 0.001). Predictors of good visual outcome included body mass index [odds ratio (OR) = 0.734, p = 0.044], serum AQP4 antibody-positive status (OR = 0.004, p = 0.001), bilaterality (OR = 0.042, p = 0.008) and time window from onset to PE therapy per 1 day (OR = 0.79, p = 0.002). This study revealed that PE treatment effectively improves the visual outcomes of patients experiencing their first attack of severe acute isolated ON after high-dose IVMP treatment. Better visual outcomes can be achieved with early PE treatment.


GABA Concentration in the Left Ventral Premotor Cortex Associates With Sensory Hyper-Responsiveness in Autism Spectrum Disorders Without Intellectual Disability.
Individuals with autism spectrum disorder (ASD) often exhibit abnormal processing of sensory inputs from multiple modalities and higher-order cognitive/behavioral response to those inputs. Several lines of evidence suggest that altered γ-aminobutyric acid (GABA), the main inhibitory neurotransmitter in the brain, is a central characteristic of the neurophysiology of ASD. The relationship between GABA in particular brain regions and atypical sensory processing in ASD is poorly understood. We therefore employed 1H magnetic resonance spectroscopy (1H-MRS) to examine whether GABA levels in brain regions critical to higher-order motor and/or multiple sensory functions were associated with abnormal sensory responses in ASD. We evaluated atypical sensory processing with a clinically-validated assessment tool. Furthermore, we measured GABA levels in four regions: one each in the primary visual cortex, the left sensorimotor cortex, the left supplementary motor area (SMA), and the left ventral premotor cortex (vPMC). The latter two regions are thought to be involved in executing and coordinating cognitive and behavioral functions in response to multisensory inputs. We found severer sensory hyper-responsiveness in ASD relative to control participants. We also found reduced GABA concentrations in the left SMA but no differences in other regions of interest between ASD and control participants. A correlation analysis revealed a negative association between left vPMC GABA and the severity of sensory hyper-responsiveness across all participants, and the independent ASD group. These findings suggest that reduced inhibitory neurotransmission (reduced GABA) in a higher-order motor area, which modulates motor commands and integrates multiple sensory modalities, may underlie sensory hyper-responsiveness in ASD.


Seasonal Variation of Orofacial Clefts.
Orofacial clefts represent the most common craniofacial malformation diagnosed at birth and may be divided into isolated cleft lip (CL), cleft lip and palate (CL/P), or isolated cleft palate (CP). The causes of orofacial clefts have long been understood to be multifactorial; however, research into the genetic and environmental factors underpinning these disorders in African populations is scant. Seasonal variation in the occurrence of orofacial clefts was investigated. Seasonal variation is defined as differences due to periodic, temporal, and external influences, namely the particular time or season of the year. The study design is a retrospective record review and sampled patients presenting to the Cleft Clinic at Charlotte Maxeke Johannesburg Academic Hospital during the time period October 2000 to October 2015. Ethical approval was obtained. Information recorded included patient demographics (date of birth and gender), diagnosis and laterality of the cleft. This study includes 512 infants. This study shows seasonal variation in the incidence of orofacial clefts limited to patients with both a CL/P. There was no significant seasonal variation observed in isolated CL or CP and laterality of the cleft. Significant seasonal variation was observed only in those born in winter versus summer, with more children born with a CL/P in winter months. Seasonal (or temporal) variation indicates a distinct environmental influence on the formation of orofacial clefts in utero, more specifically in pregnancies with the first trimester during spring. Several reasons have been advanced and include sunlight exposure and vitamin D levels, weight gain, and maternal obesity. More study is required to further elucidate these reasons.


Google Ranking of Plastic Surgeons Values Social Media Presence Over Academic Pedigree and Experience.
Patients increasingly rely on online resources to make healthcare decisions. Google dominates the search engine market; first-page results receive most of the web traffic and therefore serve as an important indicator of consumer reach. Our objective was to analyze the respective importance of physician academic pedigree, experience, and social media presence on plastic surgeon Google first-page search result placement. A Google.com search was conducted in the top 25 United States metropolitan areas to identify the top 20 websites of board-certified plastic surgeons. Social media presence was quantified by tracking the number of followers on Facebook, Twitter, and Instagram for every surgeon as well as medical school and year of graduation. The primary outcome was website ranking in the first page of Google search results. To identify the independent predictors of presence on the front page, we performed a multivariate logistic regression. Total number of social medial followers was associated with Google front-page placement (P < 0.001), whereas medical school ranking and years in practice were not (P = 0.17 and 0.39, respectively). A total 19.6% of plastic surgeon practices in our study cohort still had no social media accounts whatsoever. For the past few decades, plastic surgery practices relied on referrals, word of mouth, and the surgeon's reputation and academic pedigree to attract new patients. It is now clear that this practice-building model is being rapidly supplanted by a new paradigm based on social media presence to reach potential patients.


Heterogeneity of uridine incorporation along the rabbit nephron. II. Effect of DOCA.
Uridine uptake was investigated in microdissected segments from the entire length of tubules from normal rabbit kidneys and the effect DOCA treatment was studied. Uridine uptake was measured after in vitro incubation of kidney pyramids with [14C]uridine (0.25 mM). Determinations were done on intact segments, and also using TCA precipitation on both the acid-soluble fraction considered as representative of the pyrimidine nucleoside precursor pool and the acid-insoluble one containing the precipitated nucleic acids. In normal rabbits the fraction of uridine incorporated in the insoluble fraction represented about 5% of that in the soluble one for all segments. Maximal values were found in the distal convoluted tubule (DCT) and the connecting tubule (CNT), where uridine uptake was two- to fourfold higher than in other segments. After DOCA treatment, a significant increase in uridine uptake in the soluble fraction was present in the cortical collecting (CCT) (+90%, P less than 0.001) and medullary collecting tubules (MCT) (+63%, P less than 0.05) when values were corrected for the cellular swelling induced by DOCA in all segments. An even more marked rise in uridine incorporation was observed in the insoluble fraction of the DCT (+88%, P less than 0.05), CCT (+291%, P less than 0.001), and MCT (+422%, P less than 0.05). These results indicate that 1) the level of uridine uptake in soluble and insoluble fractions widely differs along the tubular length of the normal rabbit nephron; and 2) DOCA treatment induces an increase of uridine incorporation in both the pyrimidine precursor pool and nucleic acids of the collecting tubule, known to be the target site for corticosteroid hormones.


Characterization and dating of blue ballpoint pen inks using principal component analysis of UV-Vis absorption spectra, IR spectroscopy, and HPTLC.
The ink of pens and ink extracted from lines on white photocopier paper of 10 blue ballpoint pens were subjected to ultraviolet-visible (UV-Vis) spectroscopy, infrared (IR), and high-performance thin-layer liquid chromatography (HPTLC). The R(f) values and color tones of the bands separated by thin-layer chromatography (TLC) analysis used to classify the writing inks into three groups. The principal component analysis (PCA) investigates the pen responsible for a piece of writing, and how time affects spectroscopy of written ink. PCA can differentiate between pen ink and ink line indicates the influence of solvent extraction process on the results. The PCA loadings are useful in individualization of a questioned ink from a database. The PCA of ink lines extracted at different times can be used to estimate the time at which a questioned document was written. The results proved that the UV-Vis spectra are effective tool to separate blue ballpoint pen ink in most cases rather than IR and HPTLC.


The potential implications of a PCOS diagnosis on a woman's long-term health using data linkage.
The polycystic ovary syndrome (PCOS) is the commonest endocrine abnormality in women of reproductive age. To determine the rate of hospital admissions for women with PCOS in Western Australian population in comparison to women without PCOS. A population-based retrospective cohort study using data linkage in a statewide hospital morbidity database system. All hospitals within Western Australia. A total of 2566 women with PCOS hospitalized from 1997-2011 and 25 660 randomly selected age-matched women without a PCOS diagnosis derived from the electoral roll. Hospitalizations by ICD-10-M diagnoses from 15 years were compared. Hospitalizations were followed until a median age of 35.8 years (interquartile range, 31.0-39.9). PCOS was associated with more nonobstetric and non-injury-related hospital admissions (median, 5 vs 2; P < .001), a diagnosis of adult-onset diabetes (12.5 vs 3.8%), obesity (16.0 vs 3.7%), hypertensive disorder (3.8 vs 0.7%), ischemic heart disease (0.8 vs 0.2%), cerebrovascular disease (0.6 vs 0.2%), arterial and venous disease (0.5 vs 0.2% and 10.4 vs 5.6%, respectively), asthma (10.6 vs 4.5%), stress/anxiety (14.0 vs 5.9%), depression (9.8 vs 4.3%), licit/illicit drug-related admissions (8.8 vs 4.5%), self-harm (7.2 vs 2.9%), land transport accidents (5.2 vs 3.8%), and mortality (0.7 vs 0.4%) (all P < .001). Women with PCOS had a higher rate of admissions for menorrhagia (14.1 vs 3.6%), treatment of infertility (40.9 vs 4.6%), and miscarriage (11.1 vs 6.1%) and were more likely to require in vitro fertilization (17.2 vs 2.0%). PCOS has profound medical implications for the health of women, and health care resources should be directed accordingly.


Preservation of Simmental bull sperm at 0°C in Tris dilution: effect of dilution ratio and long-distance transport.
This study aimed to assess the impact of the dilution ratio of Tris diluent, storage at 0°C, and long-distance transportation on the spermatozoa of Simmental cattle. It also validated the feasibility of the regional distribution of fresh semen. In experiment 1, semen was diluted at four dilution ratios (1:6, 1:9, 1:12, and 1:15) to determine the optimal dilution ratio of Tris diluent. In experiment 2, we assessed sperm viability, progressive motility (objectively assessed by computer-assisted sperm analyzer), and acrosome intactness in Tris dilutions kept at constant 0°C for 1, 3, 6, 9, and 12 days. We compared them to Tianshan livestock dilutions (Commercial diluent). In experiment 3, semen was diluted using Tris diluent, and sperm quality was measured before and after long-distance transport. Artificial insemination of 177 Simmental heifers compared to 156 using Tianshan Livestock dilution. The outcomes demonstrated that 1:9 was the ideal Tris diluent dilution ratio. The sperm viability, Progressive Motility, and acrosome integrity of both Tris and Tianshan dilutions preserved at 0°C gradually decreased over time. sperm viability was above 50% for both dilutions on d 9, with a flat rate of decline. The decrease in acrosome integrity rate was faster for Tianshan livestock dilutions than for Tris dilutions when stored at 0°C for 1-6 days. There was no significant difference (p > 0.05) in sperm viability between semen preserved in Tris diluent after long-distance transportation and semen preserved in resting condition. The conception rates for Tris dilution and Tianshan livestock dilution were 49.15% and 46.15% respectively, with no significant difference (p > 0.05). This shows that Tris diluent is a good long-term protectant. It has been observed that fresh semen can be successfully preserved for long-distance transport when stored under 0°C conditions. Additionally, it is feasible to distribute semen regionally.


Autoantibodies to a novel cell cycle-regulated protein that accumulates in the nuclear matrix during S phase and is localized in the kinetochores and spindle midzone during mitosis.
We have employed human autoantibodies to characterize a novel cell cycle-regulated nuclear protein, provisionally designated p330d (doublet polypeptide of 330 kDa). The expression and intracellular distribution of this protein was followed throughout the cell cycle using immunofluorescence microscopy, laser confocal microscopy, immunoelectron microscopy and flow cytometry. p330d was expressed only in proliferating cells and began accumulating in the nucleus during early S phase. The protein reached maximum expression levels during G2/M. In situ extractions with detergent, salt and nucleases failed to abolish the nuclear staining of interphase cells, suggesting a tight binding of p330d to the nuclear matrix during interphase. p330d was concentrated in the kinetochores during prophase but was relocated to the spindle midzone at the onset of anaphase. By late telophase, it was localized predominantly in the intercellular bridge regions flanking the midbody and disappeared gradually as the daughter cells separated. Immunoblotting analysis showed that the autoimmune sera recognized a doublet of 330 kDa, and affinity-purified antibodies from this doublet reproduced the fluorescence staining pattern of the whole serum. We propose that p330d is a novel member of the class of 'chromosomal passenger' proteins, which are associated transiently with centromeres during early mitosis and are then redistributed to other sites of the mitotic apparatus after the metaphase/anaphase transition. Possible in vivo functions for p330d and related proteins might include roles in centromere/kinetochore maturation and assembly, chromosome segregation, central spindle stabilization and cytokinesis.


Non-linear and chaos characteristics of heart sound time series.
Analysis of chaotic time series is common in many fields of science and engineering. It arises primarily from massive interactions between the many different parts of a non-linear system or in non-linear physical phenomena that are intrinsically complex. It is important to analyse the time series of these non-linear dynamic systems based on chaos theory. In recent years, many researchers on heart dynamics have demonstrated that chaos really exists in heart movements. In this study the non-linear and chaos characteristics are investigated and the fractal dimensions (FDs) and largest Lyapunov exponents (LLE) of the heart sound time series are calculated. First, the C-C method is used to estimate the time delay and embedding dimensions which are used to reconstruct the phase spaces. Then, the FDs and LLEs of the different heart sound signals are calculated and analysed, including the healthy heart sound, splitting of the second heart sound, mitral incompetence, and abnormal aortic shrinkage. From the results, the non-linear and chaotic characteristics in heart dynamic movement are found, and the results of LLEs show that the healthy heart has more obvious chaotic movements than abnormal movements.


Effectiveness and Safety of Linezolid Versus Vancomycin, Teicoplanin, or Daptomycin against Methicillin-Resistant Staphylococcus aureus Bacteremia: A Systematic Review and Meta-Analysis.
Vancomycin (VCM) and daptomycin (DAP) are standard therapies for methicillin-resistant Staphylococcus aureus (MRSA) bacteremia, despite concerns regarding clinical utility and growing resistance. Linezolid (LZD) affords superior tissue penetration to VCM or DAP and has been successfully used as salvage therapy for persistent MRSA bacteremia, indicating its utility as a first-choice drug against MRSA bacteremia. In a systematic review and meta-analysis, we compared the effectiveness and safety of LZD with VCM, teicoplanin (TEIC), or DAP in patients with MRSA bacteremia. We evaluated all-cause mortality as the primary effectiveness outcome, clinical and microbiological cure, hospital length of stay, recurrence, and 90-day readmission rates as secondary effectiveness outcomes, and drug-related adverse effects as primary safety outcomes. We identified 5328 patients across 2 randomized controlled trials (RCTs), 1 pooled analysis of 5 RCTs, 1 subgroup analysis (1 RCT), and 5 case-control and cohort studies (CSs). Primary and secondary effectiveness outcomes were comparable between patients treated with LZD versus VCM, TEIC, or DAP in RCT-based studies and CSs. There was no difference in adverse event incidence between LZD and comparators. These findings suggest that LZD could be a potential first-line drug against MRSA bacteremia as well as VCM or DAP.


Influence of leptin, androgens and insulin sensitivity on increased GH response to clonidine in lean patients with polycystic ovary syndrome.
Our aim was to investigate whether insulin sensitivity, leptin, androgen or estradiol levels are associated with disturbed GH response to clonidine in lean patients with polycystic ovary syndrome. Fourteen lean polycystic ovary syndrome patients, 11 ovulatory patients presenting idiopathic hirsutism and 10 non-hirsute, normal women with regular cycles paired for age and BMI were included in a cross-sectional study. Baseline hormonal and metabolic variables were assessed and analyzed in association with GH response to oral administration of 0.3 mg of clonidine. Delta GH was significantly higher in the PCOS group than in the IH and control groups (p = 0.014). The groups were similar in terms of body mass index, insulin, glucose, total and HDL cholesterol, triglycerides and estradiol levels. Free androgen index (r = 0. 454, p = 0.015) and leptin (r = 0.419, p = 0.023) were positively correlated with the homeostasis model assessment. The homeostasis model assessment was the only variable that significantly correlated with GH response to clonidine (r = 0.375, p = 0.029) (vs. estradiol, free androgen index, leptin and LH). Nonetheless, when the analysis was adjusted for leptin levels and free androgen index, the statistical significance of this correlation was lost. The increased GH secretion observed in our lean PCOS patients may be associated with slight changes in insulin sensitivity, even in the absence of clinical evidence of insulin resistance. This association seems to be modulated by leptin and androgen levels.


Glycine immunoreactivity in the lateral nucleus of the trapezoid body of the cat.
The central auditory system contains several predominantly glycine-immunoreactive nuclei, and one of these, the lateral nucleus of the trapezoid body, contains cell bodies exhibiting a spectrum of labeling intensity. By using post-embedding glycine immunocytochemistry on thin sections, and toluidine blue staining of adjacent sections, we established that darkly glycine-immunoreactive neurons constituted a distinct morphological class and form one of three subnuclei of the lateral nucleus of the trapezoid body, called the posteroventral subnucleus. These neurons resemble, in both labeling intensity and cell body morphology, the principal cells of the medial nucleus of the trapezoid body. The other two subnuclei of the lateral nucleus of the trapezoid body, its main and hilus subnuclei, contained predominantly glycine-immunoreactive and glycine-immunonegative neurons, respectively. Glycine immunoreactivity was compared with gamma-aminobutyric acid (GABA) immunoreactivity in order to identify other organizational features of the lateral nucleus of the trapezoid body. Cell bodies that displayed either dark glycine-immunoreactivity or which were glycine-immunonegative were GABA-immunonegative. Cell bodies that displayed GABA immunoreactivity were preferentially located in the main subnucleus. Patterns of distribution of axosomatic innervation in the lateral nucleus of the trapezoid body were revealed in which glycine-immunoreactive puncta were (1) more numerous than GABA-immunoreactive puncta on glycine-immunonegative cell bodies and (2) equal to or less numerous than GABA-immunoreactive puncta on glycine-immunoreactive cell bodies. The characteristics of neural circuitry revealed by glycine and GABA immunoreactivity in the lateral nucleus of the trapezoid body may be generalizable to other populations of neurons of the superior olivary complex and to other regions of the central nervous system containing glycinergic neurons, such as the retina.


Using simulation pedagogy to enhance teamwork and communication in the care of older adults: the ELDER project.
The Expanded Learning and Dedication to Elders in the Region (ELDER) project addressed the needs of under-served older adults by educating health care providers in home health and long-term care facilities. Four agencies in a health professional shortage/medically underserved area participated. Focus groups were held to determine agency-specific educational needs. Curricula from the John A. Hartford Foundation were adapted to design unique curricula for each agency and level of personnel during the first 2 years. The focus of this report is the case-based simulation learning approach used in year 3 to validate application of knowledge and facilitate teamwork and interprofessional communication. Three simulation sessions on varying topics were conducted at each site. Postsimulation surveys and qualitative interviews with hired evaluators showed that participants found simulations helpful to their practice. Tailored on-site education incorporating mid-fidelity simulation was an effective model for translating gerontological knowledge into practice and encouraging communication and teamwork in these settings.


Changes in quantity, spending, and nutritional characteristics of adult, adolescent and child urban corner store purchases after an environmental intervention.
The purpose of this study was to assess one-year changes in corner store purchases (nutritional characteristics, amount spent) of children, adolescents, and adults in a low-income urban environment before and after implementing an environmental intervention to increase the availability of healthier products. Corner store owners were provided tools (trainings, signage, refrigeration) to increase the promotion and availability of several healthy foods. Based on the degree of support provided, stores were classified as "basic" or "high-intensity" intervention stores. Data on purchases and their nutrient content were gathered (n = 8671 at baseline, n = 5949 at follow-up) through customer purchase assessment interviews and direct observation outside of 192 corner stores in Philadelphia from March 2011 to August 2012. At baseline, shoppers spent $2.81 ± 3.52 for 643 ± 1065 kcal. Energy, select nutrients, and the total amount spent did not significantly change in the overall sample from baseline to follow-up. Similarly, there was no effect on energy and nutrient content when comparing changes over time between basic and high-intensity stores. There were no significant changes in the energy or nutrient content of corner store purchases one year after implementation of environmental changes to increase the availability of healthier products.


Serum Syndecan-1: A Novel Biomarker for Pancreatic Ductal Adenocarcinoma.
Syndecan-1 (SDC1) has multiple functions in tumorigenesis in general and specifically in pancreatic cancer. We aimed to evaluate SDC1 as a diagnostic and prognostic biomarker in patients with pancreatic ductal adenocarcinoma (PDAC). In this case-control study, patients newly diagnosed with a biopsy-proven PDAC were enrolled alongside healthy individuals in a derivation-validation cohort design. Serum SDC1 was measured by enzyme-linked immunoassay. The diagnostic accuracy of SDC1 levels for diagnosing PDAC was computed. A unified cohort enriched with additional early-stage patients with PDAC was used to evaluate the association of SDC1 with survival outcomes and patient characteristics. In the derivation cohort, serum SDC1 levels were significantly higher in patients with PDAC (n = 39) compared with healthy controls (n = 20) (40.1 ng/mL, interquartile range 29.8-95.3 vs 25.6 ng/mL, interquartile range 17.1-29.8, respectively; P < 0.001). The receiver operating characteristic analysis area under the curve was 0.847 (95% confidence interval 0.747-0.947, P < 0.001). These results were replicated in a separate age-matched validation cohort (n = 38 PDAC, n = 38 controls; area under the curve 0.844, 95% confidence interval 0.757-0.932, P < 0.001). In the combined-enriched PDAC cohort (n = 110), using a cutoff of 35 ng/mL, the median overall 5-year survival between patients below and above this cutoff was not significantly different, although a trend for better survival after 1 year was found in the lower level group (P = 0.06). There were 12 of the 110 patients with PDAC (11%) who had normal CA 19-9 in the presence of elevated SDC1. These findings suggest serum SDC1 as a promising novel biomarker for early blood-based diagnosis of pancreatic cancer.


Mode of action of beta-lactam antibiotics.
Biochemical investigations of the mode of action of beta-lactam antibiotics have focused on the interaction of these drugs with sensitive enzymes and penicillin-binding proteins in vitro and on the correlation of these data with physiological responses to the drugs. The classical response is inhibition of growth and cell death followed by lysis; however, the bacteriostatic response to penicillins, which is seen in certain bacterial species such as Streptococcus mutans and in mutants of species such as Streptococcus pneumoniae, is also described. The biochemical data remain consistent with the acyl-D-alanyl-D-alanine donor substrate analogue theory, but multiple targets with differential sensitivity to different beta-lactam antibiotics exist. The relationship of these targets to penicillin-binding proteins and their possible functions in cell growth and division are discussed.


Mortality analysis of Pleistocene bears and its paleoanthropological relevance.
Bear bones and Paleolithic stone artefacts often co-occur in Pleistocene cave deposits of Eurasia, raising the question of how these associations come about and the need for effective methods with which to obtain a clear answer. Building upon knowledge of modern bears, I present a method for testing two competing hypotheses about the causes of bear mortality in hibernation contexts. The first hypothesis proposes that age-dependent deaths resulted from non-violent causes (principally starvation), implying that bears' presence in a cave was not linked in time to human activities there. The second hypothesis proposes that random bear deaths in caves resulted from hunting by humans or other large predators, implying a temporal link between them; the expectation of a nonselective age pattern in this circumstance arises from the fact that the individual characters of hibernating bears are hidden from predators. Three elements of the method and its development are presented: (1) a brief review of the biological bases of hibernation-related mortality in modern Ursus, its paleontological consequences, and test expectations drawn therefrom; (2) a detailed, illustrated technique for age-scoring isolated bear cheek teeth based on tooth eruption-wear sequences, developed primarily for cave and brown bears; and, (3) a simple, accurate way to evaluate real cases in terms of contrasting mortality models. The final step is demonstrated by application to a Middle Pleistocene cave bear assemblage (Ursus deningeri) from Yarimburgaz Cave in Turkey, a large collection found in general stratigraphic association with Paleolithic artefacts. The advantages of the method include its ability to (a) handle small samples, (b) use isolated tooth specimens, and (c) evaluate cases simultaneously in terms of idealized age structure models and the variation that normally is associated with each under natural conditions. While the more obvious benefit of bear mortality analysis may be to research on ancient bear demography, the principles and procedures offered here are equally pertinent to archaeological studies of carnivore-mediated formation processes in cave sites. As is generally true in taphonomic research, however, bear mortality patterns are most effective when used in combination with independent lines of evidence to address questions about assemblage formation.


Mechanics of epithelial closure over non-adherent environments.
The closure of gaps within epithelia is crucial to maintain its integrity during biological processes such as wound healing and gastrulation. Depending on the distribution of extracellular matrix, gap closure occurs through assembly of multicellular actin-based contractile cables or protrusive activity of border cells into the gap. Here we show that the supracellular actomyosin contractility of cells near the gap edge exerts sufficient tension on the surrounding tissue to promote closure of non-adherent gaps. Using traction force microscopy, we observe that cell-generated forces on the substrate at the gap edge first point away from the centre of the gap and then increase in the radial direction pointing into the gap as closure proceeds. Combining with numerical simulations, we show that the increase in force relies less on localized purse-string contractility and more on large-scale remodelling of the suspended tissue around the gap. Our results provide a framework for understanding the assembly and the mechanics of cellular contractility at the tissue level.


Influence of water and soil temperature on the concentration and efficacy of phenamiphos for control of root-knot nematodes.
Field plots of Tifton loamy sand were treated with phenamiphos for control of root-knot nematodes in a multiple-crop system of turnips, field corn, and southern peas. Preplant applications of phenamiphos protected roots of turnips and corn from damage by root-knot nematodes. Concentrations of phenamiphos at application in the 0-15-cm soil layer were near 6 mug/g on turnips and near 4 mug/g on corn and southern peas. After 30 d, concentrations were approximately 1 mug phenamiphos/g of soil for all crops. Concentrations of 2.0-3.8 mug phenamiphos/g of soil for 9-d duration appeared to be adequate for control of root-knot nematodes on field corn and southern peas in this multiple-crop system. Stepwise regression analyses indicated that 31%, 62%, and 22% of the variations in concentration of phenamiphos in the soil planted to turnips, corn, and southern peas, respectively, were attributable to the amount of water that the plots received. Soil temperature had no effect on concentrations of phenamiphos.


Pharmacokinetics of colistin methanesulfonate and formed colistin in end-stage renal disease patients receiving continuous ambulatory peritoneal dialysis.
Colistin, administered intravenously as its inactive prodrug colistin methanesulfonate (CMS), is increasingly used as last-line therapy to combat multidrug-resistant Gram-negative bacteria. CMS dosing needs to be adjusted for renal function. The impact of continuous ambulatory peritoneal dialysis (CAPD) on the pharmacokinetics of both CMS and colistin has not been studied. No CMS dosing recommendations are available for patients receiving CAPD. Eight CAPD patients received a single intravenous CMS dose (150 mg colistin base activity [CBA]) over 30 min. Serial blood and dialysate samples, and cumulative urine where applicable, were collected over 25 h. CMS and colistin concentrations were determined by high-performance liquid chromatography. Population pharmacokinetic modeling and Monte Carlo simulations were conducted. The total body clearance of CMS (excluding CAPD clearance) was 1.77 liters/h (44%) [population mean (between-subject variability)], while CAPD clearance was 0.088 liter/h (64%). The population mean terminal half-life of CMS was 8.4 h. For colistin, the total clearance/fraction of CMS metabolized to colistin (fm) (excluding CAPD clearance) was 2.74 liters/h (50%), the CAPD clearance was 0.101 liter/h (34%), and the mean terminal half-life was 13.2 h. Monte Carlo simulations suggested a loading dose of 300 mg CBA on day 1 and a maintenance dose of either 150 mg or 200 mg CBA daily to achieve a target average steady-state plasma colistin concentration of 2.5 mg/liter. Clearance by CAPD was low for both CMS and formed colistin. Therefore, CMS doses should not be increased during CAPD. Modeling and simulation enabled us to propose the first evidence-based CMS dosage regimen for CAPD patients.


Effect of solvent type on microtensile bond strength of a total-etch one-bottle adhesive system to moist or dry dentin.
This study evaluated the effect of organic solvent (acetone or ethanol) on the microtensile bond strengths (MTBS) of an adhesive system applied to dry and moist dentin. Sixteen extracted human third molars were ground to expose a flat occlusal dentin surface and acid etched for 20 seconds (20% phosphoric acid gel, Gluma Etch 20 Gel, Heraeus/Kulzer). After rinsing the acid etchant, an ethanol-based one-bottle adhesive system was applied to the mesial half of the occlusal dentin surface. An acetone-based, one-bottle adhesive system was applied to the distal half of the ground dentin surface. The teeth were randomly assigned to groups. In Group 1, the etched dentin was thoroughly air dried and an ethanol-based one-bottle adhesive system was applied (Gluma Comfort Bond, Heraeus/Kulzer) (GCB). In Group 2, the etched dentin was thoroughly air dried and an acetone-based one-bottle adhesive system was applied (Gluma One Bond, Heraeus/Kulzer)(GOB). In Group 3, excess moisture was removed after acid etching, leaving a moist dentin surface and a one-bottle ethanol-based adhesive was applied (Gluma Comfort Bond). In Group 4, excess moisture was removed after acid etching, leaving a moist dentin surface and an acetone-based adhesive was applied (Gluma One Bond). A hybrid resin composite (Venus, Heraeus/Kulzer) was applied to the bonded surface in four 1-mm increments and light cured according to manufacturer's directions. The specimens were then sectioned with a slow-speed diamond saw in two perpendicular directions to obtain sticks with a cross-section of 0.5 +/- 0.05 mm2. The microtensile bond strength (MTBS) test was performed with a Bencor device in an Instron machine at a crosshead speed of 0.5 mm/minute. The data were subjected to a two-way ANOVA and Scheffé Post hoc test (p < 0.05). The experimental MTBS measured for dry dentin were Group 1 = 37.0 +/- 10.6 and Group 2 = 34.7 +/- 9.0 in MPa (mean +/- SD); and on moist dentin, Group 3 = 50.7 +/- 11.0 and Group 4 = 38.5 +/- 10.5 in MPa (mean +/- SD). The ethanol based adhesives resulted in higher MTBS than acetone-based adhesive (p < 0.008) and bonding to moist dentin resulted in higher MTBS (p < 0.001). GCB applied on moist dentin resulted in statistically higher bond strengths than the other groups. The highest MTBS were achieved with the use of an ethanol-based adhesive to moist dentin.


Clusters and the new economics of competition.
Economic geography in an era of global competition poses a paradox. In theory, location should no longer be a source of competitive advantage. Open global markets, rapid transportation, and high-speed communications should allow any company to source any thing from any place at any time. But in practice, Michael Porter demonstrates, location remains central to competition. Today's economic map of the world is characterized by what Porter calls clusters: critical masses in one place of linked industries and institutions--from suppliers to universities to government agencies--that enjoy unusual competitive success in a particular field. The most famous example are found in Silicon Valley and Hollywood, but clusters dot the world's landscape. Porter explains how clusters affect competition in three broad ways: first, by increasing the productivity of companies based in the area; second, by driving the direction and pace of innovation; and third, by stimulating the formation of new businesses within the cluster. Geographic, cultural, and institutional proximity provides companies with special access, closer relationships, better information, powerful incentives, and other advantages that are difficult to tap from a distance. The more complex, knowledge-based, and dynamic the world economy becomes, the more this is true. Competitive advantage lies increasingly in local things--knowledge, relationships, and motivation--that distant rivals cannot replicate. Porter challenges the conventional wisdom about how companies should be configured, how institutions such as universities can contribute to competitive success, and how governments can promote economic development and prosperity.


Probing Projections: Interaction Techniques for Interpreting Arrangements and Errors of Dimensionality Reductions.
We introduce a set of integrated interaction techniques to interpret and interrogate dimensionality-reduced data. Projection techniques generally aim to make a high-dimensional information space visible in form of a planar layout. However, the meaning of the resulting data projections can be hard to grasp. It is seldom clear why elements are placed far apart or close together and the inevitable approximation errors of any projection technique are not exposed to the viewer. Previous research on dimensionality reduction focuses on the efficient generation of data projections, interactive customisation of the model, and comparison of different projection techniques. There has been only little research on how the visualization resulting from data projection is interacted with. We contribute the concept of probing as an integrated approach to interpreting the meaning and quality of visualizations and propose a set of interactive methods to examine dimensionality-reduced data as well as the projection itself. The methods let viewers see approximation errors, question the positioning of elements, compare them to each other, and visualize the influence of data dimensions on the projection space. We created a web-based system implementing these methods, and report on findings from an evaluation with data analysts using the prototype to examine multidimensional datasets.


3,4-dihydroxytoluene, a metabolite of rutin, suppresses the progression of nonalcoholic fatty liver disease in mice by inhibiting p300 histone acetyltransferase activity.
3,3',4',5,7-Pentahydroxyflavone-3-rhamnoglucoside (rutin) is a flavonoid with a wide range of pharmacological activities. Dietary rutin is hardly absorbed because the microflora in the large intestine metabolize rutin into a variety of compounds including quercetin and phenol derivatives such as 3,4-dihydroxyphenolacetic acid (DHPAA), 3,4-dihydroxytoluene (DHT), 3,4-hydroxyphenylacetic acid (HPAA) and homovanillic acid (HVA). We examined the potential of rutin and its metabolites as novel histone acetyltransferase (HAT) inhibitors. DHPAA, HPAA and DHT at the concentration of 25 μM significantly inhibited in vitro HAT activity with DHT having the strongest inhibitory activity. Furthermore, DHT was shown to be a highly efficient inhibitor of p300 HAT activity, which corresponded with its high degree of inhibition on intracellular lipid accumulation in HepG2 cells. Docking simulation revealed that DHT was bound to the p300 catalytic pocket, bromodomain. Drug affinity responsive target stability (DARTS) analysis further supported the possibility of direct binding between DHT and p300. In HepG2 cells, DHT concentration-dependently abrogated p300-histone binding and induced hypoacetylation of histone subunits H3K9, H3K36, H4K8 and H4K16, eventually leading to the downregulation of lipogenesis-related genes and attenuating lipid accumulation. In ob/ob mice, administration of DHT (10, 20 mg/kg, iv, every other day for 6 weeks) dose-dependently improved the NAFLD pathogenic features including body weight, liver mass, fat mass, lipid accumulation in the liver, and biochemical blood parameters, accompanied by the decreased mRNA expression of lipogenic genes in the liver. Our results demonstrate that DHT, a novel p300 histone acetyltransferase inhibitor, may be a potential preventive or therapeutic agent for NAFLD.


Separation of peptides from detergents using ion mobility spectrometry.
Mass spectrometry (MS) has dramatically evolved in the last two decades and has been the driving force of the spectacular expansion of proteomics during this period. However, the very poor compatibility of MS with detergents is still a technical obstacle in some studies, in particular on membrane proteins. Indeed, the high hydrophobicity of membrane proteins necessitates the use of detergents for their extraction and solubilization. Here, we address the analytical potential of high-field asymmetric waveform ion mobility spectrometry (FAIMS) for separating peptides from detergents. The study was focused on peptides from the human integral membrane protein CD9. A tryptic peptide was mixed with the non-ionic detergents Triton X-100 or beta-D-dodecyl maltoside (DDM) as well as with the ionic detergents sodium dodecyl sulfate (SDS) or sodium deoxycholate (SDC). Although electrospray ionization (ESI) alone led to a total suppression of the peptide ion signal on mass spectra with only detection of the detergents, use of FAIMS allowed separation and clear identification of the peptide with any of the detergents studied. The detection and identification of the target compound in the presence of an excess of detergents are then feasible. FAIMS should prove especially useful in the structural and proteomic analysis of membrane proteins.


Kinetics of aggregation and crystallization of polyaspartic Acid stabilized calcium phosphate particles at high concentrations.
Bone is an important material to study due to its exceptional mechanical properties and relevance with respect to hard tissue regeneration and repair. A significant effort has been directed toward understanding the bone formation process and the production of synthetic bone mimicking materials. Here, the formation and structural evolution of calcium phosphate (CaP) was investigated in the presence of relatively high concentrations of calcium, phosphate, and polyaspartic acid (pAsp) using dynamic light scattering (DLS) and cryo-transmission electron microscopy (cryo-TEM). The incipient CaP aggregates were comprised of spherical nanoparticles (diameter ≈ 3-4 nm); they became preferentially aligned over time and eventually transformed into nanorods. The nanorods remained stable in suspension with no signs of further aggregation for at least four months. Detailed cryo-TEM suggested that the CaP nanorods formed through an oriented attachment mechanism. These results show that the reaction concentration greatly influences the mechanism and final properties of CaP. Mechanistic insights gained from this study will facilitate better design and fabrication of bioinspired materials.


Brazilian biologic registry: BiobadaBrasil implementation process and preliminary results.
The present study aimed at describing the implementation process of a national registry in a developing country (Brazil) and at reporting the main preliminary results of the BiobadaBrasil registry. Through a PANLAR agreement, the Biobadaser protocol was used as a model for implementing the new registry in our country. During the first two years of this effort, the original protocol was adapted, translated, and presented to all Brazilian rheumatologists. For ten months, data of 1,037 patients (750 subjects treated with biological drugs and 287 control subjects) from 15 centers were collected. Most patients had rheumatoid arthritis (RA) (n = 723). Infliximab was the most frequently used anti-TNF agent, and the total exposure to biologic drugs was 2,101 patient-years. The most common reason for interrupting drug use was lack or loss of efficacy (50%), while 30% withdrew from the treatment arm due to adverse events. Three cases of tuberculosis were observed in the biologic group, with an incidence higher than that of the general Brazilian population. Infections were observed in 23% of the biologic group, and the upper respiratory tract was the most commonly affected site. Only one case of tuberculoid leprosy was observed. No deaths or malignancies attributed to drug effects were observed as of February 2010. The implementation of the BiobadaBrasil registry was successful, and, although recent, the registry has provided important data.


Antiviral and Cytotoxic Activity of Different Plant Parts of Banana (Musa spp.).
Chikungunya and yellow fever virus cause vector-borne viral diseases in humans. There is currently no specific antiviral drug for either of these diseases. Banana plants are used in traditional medicine for treating viral diseases such as measles and chickenpox. Therefore, we tested selected banana cultivars for their antiviral but also cytotoxic properties. Different parts such as leaf, pseudostem and corm, collected separately and extracted with four different solvents (hexane, acetone, ethanol, and water), were tested for in vitro antiviral activity against Chikungunya virus (CHIKV), enterovirus 71 (EV71), and yellow fever virus (YFV). Extracts prepared with acetone and ethanol from leaf parts of several cultivars exhibited strong (EC50 around 10 μg/mL) anti-CHIKV activity. Interestingly, none of the banana plant extracts (concentration 1-100 µg/mL) were active against EV71. Activity against YFV was restricted to two cultivars: Namwa Khom-Pseudostem-Ethanol (5.9 ± 5.4), Namwa Khom-Corm-Ethanol (0.79 ± 0.1) and Fougamou-Corm-Acetone (2.5 ± 1.5). In most cases, the cytotoxic activity of the extracts was generally 5- to 10-fold lower than the antiviral activity, suggesting a reasonable therapeutic window.


Venous Thromboembolism Prophylaxis in Foot and Ankle Surgery: A Worldwide Survey.
Current recommendations on thromboprophylaxis for foot and ankle (FA) surgery are often inconsistent and generally based on weak evidence. The aim of this survey study was to evaluate the current practice among orthopedic surgeons regarding venous thromboembolism (VTE) prophylaxis following FA surgery. From February 2019 to March 2020, an online questionnaire was sent by e-mail to orthopedic societies across the world. The questionnaire was hosted by the International Society of Thrombosis and Haemostais RedCAP platform. Topics of interest were VTE rates following FA surgery, duration and type of thromboprophylaxis, bleeding complications, VTE risk factors for prophylaxis and use of risk assessment. A total of 693 FA orthopedic surgeons from all continents completed the survey of whom 392 (57%) performed more than 200 FA procedures per year. A total of 669/693 (97%) respondents stated that thromboprophylaxis is necessary in FA surgeries. When thromboprophylaxis was prescribed, half of surgeons prescribed it for the duration of immobilization. Acetylsalicylic acid, low molecular weight heparin and direct-oral anticoagulants were, in this order, the preferred choice. Acetylsalicylic acid and low molecular weight heparin were predominantly prescribed in North America and Europe, respectively. Previous deep vein thrombosis, immobility, obesity and inherited thrombophilia were considered the main risk factors indicative of thromboprophylaxis use. In this survey, most surgeons agree that thromboprophylaxis is indicated for FA surgery, but the prescription, type and duration of prophylaxis differs greatly with a large intercontinental discrepancy. These survey results could be a foundation for developing uniform guidelines to optimize thromboprophylactic strategies in FA procedures around the world.


The proregion of cathepsin L is required for proper folding, stability, and ER exit.
To investigate the role of the proregion in the biosynthesis and trafficking of mouse cathepsin L, cathepsin L cDNAs encoding proteins with altered proregions were constructed and their expression in COS cells was examined. As in transformed cells, normal mouse cathepsin L was secreted by COS cells. In contrast, two altered proregion cathepsin L proteins, one in which the proregion was deleted and a second in which the proregion was replaced with that of a homologous protein (aleurain), were retained within the cell and degraded over a period of 2-6 h. Immunofluorescence localization and the lack of effect of NH4Cl and brefeldin A on the turnover of the altered cathepsin L proteins indicated that their degradation occurred in the endoplasmic reticulum (ER). By using brefeldin A to induce colocalization of the UDPGlcNAc: lysosomal enzyme N-acetylglucosamine-1-phosphotransferase with the cathepsin L proteins in the ER, it was shown that the altered proteins were not susceptible to mannose phosphorylation as they exist in the ER. Trypsin sensitivity assays indicated that altered proregion proteins synthesized in COS cells or in vitro are misfolded. Taken together, these results indicate that the proregion plays an essential role in proper folding of cathepsin L. ER retention, decreased stability, and lack of mannose phosphorylation of the altered proteins are most likely secondary effects resulting from improper folding.


The swimbladder nematode Anguillicola crassus in the European eel Anguilla anguilla and the Japanese eel Anguilla japonica: differences in susceptibility and immunity between a recently colonized host and the original host.
The swimbladder nematode Anguillicola crassus originates from Asia where it is a parasite of the Japanese eel Anguilla japonica. After its introduction to Europe about 25 years ago, the parasite spread rapidly within the indigenous populations of the European eel Anguilla anguilla and subsequently the prevalence and mean intensity appeared to stabilize. Under experimental and aquaculture conditions the naïve new host appears to be more susceptible to A. crassus compared to the original host. Both eel species develop a immune response against A. crassus. The antibody response is well characterized for the European eel, but poorly characterized for the Japanese eel. It remains unclear if antibodies have any protective function against A. crassus. Encapsulation of larvae of A. crassus can be observed in naturally infected European eels. However, encapsulation of larvae following experimental infection has not been detected in European eels, but only in Japanese eels. Reinfection experiments and intraperitoneal injection of A. crassus homogenates failed to demonstrate the development of acquired immunity in European eels. Immunization with irradiated third stage larvae provided preliminary evidence for acquired immunity against A. crassus in the Japanese eel, but not in the European eel.


Role of megalin in renal handling of aminoglycosides.
The role of megalin in tissue distribution of aminoglycosides was examined in normal rats and maleate-treated rats that shed megalin from the renal brush-border membrane. In normal rats, amikacin administered intravenously accumulated most abundantly in the renal cortex, followed by the renal medulla. No amikacin was detected in other tissues. Tissue distributions of amikacin were well correlated with megalin levels in each tissue. Bolus administration of gentamicin increased urinary excretion of megalin ligands (vitamin D binding protein and calcium), suggesting the competition between gentamicin and these megalin ligands in renal tubules. Ligand blotting showed that binding of (45)Ca(2+) to megalin was inhibited by aminoglycosides. Both megalin levels and amikacin accumulation in renal cortex were decreased by maleate injection. Then, amikacin accumulation recovered proportionate to megalin levels. These findings suggest that megalin is involved in the renal cortical accumulation of aminoglycosides in vivo. In addition, the interaction between aminoglycosides and calcium in the kidney may be due to the competition among these compounds to bind to megalin.


Compliance with stimulant medications in patients with narcolepsy.
The goals of this descriptive study were to determine what percentage of our treated narcoleptic subjects took their stimulant medications as prescribed and to examine the relationship between compliance and response to stimulant medications. Data obtained from a screening questionnaire, sleep diaries, and medical records showed that 22 of our 43 treated narcoleptic subjects reduced their dosage of stimulant medications or had not taken any stimulant medications during a 24-hour monitoring period during which they were expected to be on medication. Although we had expected better compliance among subjects who responded to stimulant medications (day-wake subjects), statistical testing revealed no significant differences between the two groups. Nor were there any significant differences in age, gender, or educational level when compliant and noncompliant subjects were compared. Only the type of drug prescribed (short versus long-acting stimulant) affected compliance; 39.4% of the subjects with prescriptions for dextroamphetamine or methylphenidate took the amount of medication prescribed compared to 87.5% of the subjects with prescriptions for pemoline.


Protective effects of riboflavin and selenium on brain microsomal Ca2+-ATPase and oxidative damage caused by glyceryl trinitrate in a rat headache model.
Migraine headaches are considered to be associated with increased mitochondrial energy metabolism. Mitochondrial oxidative stress is also important in migraine headache pathophysiology although riboflavin and selenium (Se) induced a modulator role on mitochondrial oxidative stress in the brain. The current study aimed to determine the effects of Se with/without riboflavin on the microsomal membrane Ca(2+)-ATPase (MMCA), lipid peroxidation, antioxidant, and electroencephalography (EEG) values in glyceryl trinitrate (GTN)-induced brain injury rats. Thirty-two rats were randomly divided into four groups. The first group was used as the control, and the second group was the GTN group. Se and Se plus oral riboflavin were administered to rats constituting the third and fourth groups for 10 days prior to GTN administration. The second, third, and fourth groups received GTN to induce headache. Ten hours after the administration of GTN, the EEG records and brain cortex samples were obtained for all groups. Brain cortex microsomes were obtained from the brain samples. The brain and microsomal lipid peroxidation levels were higher in the GTN group compared to the control group, whereas they were decreased by selenium and selenium + riboflavin treatments. Vitamin A, vitamin C, vitamin E, and reduced glutathione (GSH) concentrations of the brain and MMCA, GSH and glutathione peroxidase values of microsomes were decreased by the GTN administration, although the values and β-carotene concentrations were increased by Se and Se + riboflavin treatments. There was no significant change in EEG records of the four groups. In conclusion, Se with/without riboflavin administration protected against GTN-induced brain oxidative toxicity by inhibiting free radicals and the modulation of MMCA activity and supporting the antioxidant redox system.


Design, synthesis, and evaluation of a mitoxantrone probe (MXP) for biological studies.
Mitoxantrone (MX) is a robust chemotherapeutic with well-characterized applications in treating certain leukemias and advanced breast and prostate cancers. The canonical mechanism of action associated with MX is its ability to intercalate DNA and inhibit topoisomerase II, giving it the designation of a topoisomerase II poison. Years after FDA approval, investigations have unveiled novel protein-binding partners, such as methyl-CpG-binding domain protein (MBD2), PIM1 serine/threonine kinase, RAD52, and others that may contribute to the therapeutic profile of MX. Moreover, recent proteomic studies have revealed MX's ability to modulate protein expression, illuminating the complex cellular interactions of MX. Although mechanistically relevant, the differential expression across the proteome does not address the direct interaction with potential binding partners. Identification and characterization of these MX-binding cellular partners will provide the molecular basis for the alternate mechanisms that influence MX's cytotoxicity. Here, we describe the design and synthesis of a MX-biotin probe (MXP) and negative control (MXP-NC) that can be used to define MX's cellular targets and expand our understanding of the proteome-wide profile for MX. In proof of concept studies, we used MXP to successfully isolate a recently identified protein-binding partner of MX, RAD52, in a cell lysate pulldown with streptavidin beads and western blotting. An 8-step synthesis was used to generate a biotinylated-mitoxantrone probe (MXP).A pulldown of MXP demonstrated selectivity for RAD52, but not Replication Protein A.Western blot confirmed the identity of the isolated protein, RAD52.


Phenotypic and genetic relationship between linear functional type traits and milk yield for five breeds.
Lactation records of cows first calving between 18 and 35 mo were combined with linear type ratings assigned during the same lactation if before 43 mo. Phenotypic relationships were examined between final score and 13 type appraisal traits and first lactation milk yield from 2935 Ayrshire, 3154 Brown Swiss, 13,110 Guernsey, 50,422 Jersey, and 924 Milking Shorthorn records. Most phenotypic correlations between type and milk yield were low. Linear correlations of final score with first lactation milk yield were .18 to .38. Of the linear functional type traits, correlations with first lactation yield had greatest absolute values for dairy character (.19 to .53), udder depth (-.26 to -.30), and rear udder width (.20 to .31). Multiple correlations of all type appraisal traits with first lactation milk yield ranged from .41 to .59. Herd-year-season components of variance averaged 25% for type traits. Herd-year-season with sire interaction averaged 4%. Heritability estimates for final score from paternal half-sib analysis were from .11 to .21. Heritability estimates for linear traits ranged from .01 to .37. Genetic correlation between milk and final score was positive for Guernseys (.25) and Jerseys (.21). Genetic correlations between yield and most linear type traits were low to moderate except for dairy character (.53 to .77).


MNNG-induced partial phenotypic reversion of Mer- cells.
The effect of pretreatment with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) on MNNG sensitivity of the surviving population was compared in two HeLa lines, one of the Mer+ phenotype (HeLa S3) and one of the Mer- phenotype (HeLa MR). Whereas MNNG pretreatment of HeLa Mer+ cells had no effect on the MNNG sensitivity of surviving cells, Mer- cells surviving a first exposure to MNNG became much more resistant to MNNG. Comparison of the sensitivity of individual HeLa MR clones with their MNNG-pretreated population and analysis of the composition of the pretreated population showed that the majority of cells surviving the MNNG pretreatment now displayed the Mer+ phenotype in respect to sensitivity to MNNG. One MNNG-resistant clone derived from a pretreated HeLa MR population (Cl 4) was characterized further. It had a similar sensitivity to the Mer+ line to all monofunctional alkylating agents, but was as sensitive as the Mer- line to the crosslinking agent chloroethylnitrosourea. Cl 4 cells, like the Mer- cells, did not repair O6-methylguanine (O6MeG). The results suggest that the two characteristics which are usually coupled with the Mer- phenotype--lack of O6MeG repair and hypersensitivity to MNNG--can be separated.


Surrogate versus couple therapy in vaginismus.
Women who do not have a cooperative partner cannot complete the usual therapeutic process in the treatment of vaginismus, because they cannot progress to the stage of practicing the insertion of the man partner's fingers and the insertion of a penis. To compare traditional couple therapy with therapy utilizing a surrogate partner. The study was controlled and retrospective. Data were obtained from the treatment charts of patients who had come to the clinic for treatment of vaginismus. Sixteen vaginismus patients who were treated with a man surrogate partner were compared with 16 vaginismus patients who were treated with their own partners. Successful pain-free intercourse upon completion of therapy. One hundred percent of the surrogate patients succeeded in penile-vaginal intercourse compared with 75% in the couples group (P = 0.1). All surrogate patients ended the therapy because it was fully successful, compared with 69% in the couples group. Twelve percent of the couples group ended the therapy because it failed, and 19% because the couples decided to separate. Treating vaginismus with a man surrogate partner was at least as effective as couple therapy. Surrogate therapy may be considered for vaginismus patients who have no cooperative partner.


COPD Exacerbation-Related Pathogens and Previous COPD Treatment.
We evaluated whether the pathogens identified during acute exacerbation of chronic obstructive pulmonary disease (AE-COPD) are associated with the COPD medications used in the 6 months before AE-COPD. We collected the medical records of patients diagnosed with AE-COPD at 28 hospitals between January 2008 and December 2019 and retrospectively analyzed them. Microorganisms identified at the time of AE-COPD were analyzed according to the use of inhaled corticosteroid (ICS) and systemic steroid after adjusting for COPD severity. We evaluated 1177 patients with AE-COPD and available medication history. The mean age of the patients was 73.9 ± 9.2 years, and 83% were males. The most frequently identified bacteria during AE-COPD were Pseudomonas aeruginosa (10%), followed by Mycoplasma pneumoniae (9.4%), and Streptococcus pneumoniae (5.1%), whereas the most commonly identified viruses were rhinovirus (11%) and influenza A (11%). During AE-COPD, bacteria were more frequently identified in the ICS than non-ICS group (p = 0.009), and in the systemic steroid than non-systemic steroid group (p < 0.001). In patients who used systemic steroids before AE-COPD, the risk of detecting Pseudomonas aeruginosa was significantly higher during AE-COPD (OR 1.619, CI 1.007−2.603, p = 0.047), but ICS use did not increase the risk of Pseudomonas detection. The risk of respiratory syncytial virus (RSV) detection was low when ICS was used (OR 0.492, CI 0.244−0.988, p = 0.045). COPD patients who used ICS had a lower rate of RSV infection and similar rate of P. aeruginosa infection during AE-COPD compared to patients who did not use ICS. However, COPD patients who used systemic steroids within 6 months before AE-COPD had an increased risk of P. aeruginosa infection. Therefore, anti-pseudomonal antibiotics should be considered in patients with AE-COPD who have used systemic steroids.


Genetic variants in the noncoding region of RPS19 gene in Diamond-Blackfan anemia: potential implications for phenotypic heterogeneity.
Mutations in the RPS19 gene have been identified in 25% of individuals affected by Diamond-Blackfan anemia (DBA), a congenital erythroblastopenia characterized by an aregenerative anemia and a variety of malformations. More than 60 mutations in the five coding exons of RPS19 have been described to date. We previously reported a mutation (c.-1 + 26G>T) and an insertion at -631 upstream of ATG (c.-147_-146insGCCA) in the noncoding region. Because DBA phenotype is extremely heterogeneous from silent to severe and because haploinsufficiency seems to play a role in this process, it is likely that genetic variations in the noncoding regions affecting translation of RPS19 can modulate the phenotypic expression of DBA. However, to date, very few studies have addressed this question comprehensively. In this study, we performed detailed sequence analysis of the RPS19 gene in 239 patients with DBA and 110 of their relatives. We found that 6.2% of the patients with DBA carried allelic variations upstream of ATG: 3.3% with c.-1 + 26G>T; 2.5% with c.-147_-146insGCCA; and 0.4% with c.-174G>A. Interestingly, the c.-147_-146insGCCA, which has been found in a black American and French Caribbean control population, was not found in 500 Caucasian control chromosomes we studied. However, it was found in association with the same haplotype distribution of four intronic polymorphisms in our patients with DBA. Although a polymorphism, the frequency of this variant in the patients with DBA and its association with the same haplotype raises the possibility that this polymorphism and the other genetic variations in the noncoding region could play a role in DBA pathogenesis.


Pediatric brain tumors express multiple receptor tyrosine kinases including novel cell adhesion kinases.
We have used the polymerase chain reaction to clone and characterize growth factor receptor tyrosine kinases (RTKs) expressed in 3 pathologically distinct pediatric brain tumors, an anaplastic ependymoma, a glioblastoma multiforme and a primitive neuroectodermal tumor (PNET). These neoplasms are presumed to be derived from embryonic neuroepithelial precursor cells of the central nervous system. This cloning demonstrated expression of 24 distinct kinase genes: 16 receptor type kinases and 8 nonreceptor type kinases. The expression of 6 receptors, including Hek2, IRR, Ryk, FGFR3, and 2 members of the newly identified cell adhesion kinase receptor family, DDR and TKT, in such tumors has not been reported previously. Northern analysis of mRNA levels revealed DDR expression in 6 of 7 pediatric brain tumors including an ependymoma, PNET, glioblastoma and astrocytoma, and also in an adult pheochromocytoma. Thus, the DDR cell adhesion kinase may be widely expressed in pediatric brain tumors. Also, PCR cloning may be an effective procedure for characterizing RTKs in clinical tissue samples and revealing the expression of novel RTK species.


[The structural and functional classification and evolution of cytokines].
According to the type of secondary structure, cytokines are classified into three categories: alpha-spiral (IFNs-alpha, beta, omega, gamma; ILS-2, 3,4,5,6,7,9; CSFs-G, M, GM, MGF, PDGF), beta-structural (ILs-1 alpha, beta, TNFs-alpha, beta, FGF) and (alpha + beta)-structural proteins (IL-8, IFN-gamma IP-10, PF-4, bTG, GRO, 9E3). According to the type of tertiary structure, alpha-spiral proteins are grouped into IFN- and IL-2-like families and beta-structural ones into IL-1-, and TNF-like families. Two subfamilies can be identified in the IFN-like family. Theoretical and experimental evidence suggests that the genes IFNs are products of divergent or convergent evolution towards the gene of the ancient intracellular protein alpha-prothymosine, which is evolutionally in turn associated with the L7/I1 protein of two ribosomes. It is suggested that the proteins of the immunoglobulin superfamily, including cytokine receptors descended from the ancient proteins of the unicellular organisms molecular shaperons.


Intravenous infusion route in maternal resuscitation: a scoping review.
The concept that upper extremities can be used as an infusion route during cardiopulmonary resuscitation in pregnant women is a reasonable recommendation considering the characteristic circulation of pregnant women; however, this method is not based on scientific evidence. We conducted a scoping review to determine whether the infusion route should be established above the diaphragm during cardiopulmonary resuscitation in a pregnant woman. We included randomized controlled trials (RCTs) and non-RCTs on the infusion of fluids in pregnant women after 20 weeks of gestation requiring establishment of an infusion route due to cardiac arrest, massive bleeding, intra-abdominal bleeding, cesarean section, severe infection, or thrombosis. In total, 3150 articles from electronic database were extracted, respectively. After title and abstract review, 265 articles were extracted, and 116 articles were extracted by full-text screening, which were included in the final analysis. The 116 articles included 78 studies on infusion for pregnant women. The location of the intravenous infusion route could be confirmed in only 17 studies, all of which used the upper extremity to secure the venous route. Pregnant women undergo significant physiological changes that differ from those of normal adults, because of pressure and drainage of the inferior vena cava and pelvic veins by the enlarged uterus. Therefore, despite a lack of evidence, it seems logical to secure the infusion route above the diaphragm when resuscitating a pregnant woman.


Shift of symbiont communities in Acropora tenuis juveniles under heat stress.
Ocean warming is a major threat to coral reefs, leading to an increasing frequency and amplitude of coral bleaching events, where the coral and its algal symbiont associations breakdown. Long-term change and resilience of a symbiont community in coral juveniles is thought to be one of the most important aspects for determining thermal tolerance of the coral holobionts; however, despite its importance, they are not well documented in both under elevated temperature and even under natural condition. Here we investigated changes in symbiont communities in juveniles of the coral Acropora tenuis under controlled heat stress conditions (30 °C, 31/32 °C) and natural variations in seawater temperatures (19-30 °C) for up to four months. Compared with the ambient temperature conditions, coral survival rates were higher when exposed to 30 °C, but survival rates decreased when exposed to 31/32 °C. Symbiodinium types A3, C1, and D1-4 were detected in the juveniles under all thermal conditions; however, in higher water temperatures (31/32 °C), both the prevalence of D1-4 Symbiodinium and the number of juveniles harboring only this type of symbiont increased after two to four months later. In contrast, colonies at lower temperatures (30 °C and ambient) harbored multiple clades of symbionts over the same experimental period. These results highlight the flexibility of the coral-Symbiodinium symbiosis for juvenile A. tenuis under variable thermal conditions. In particular, the benefit of the preferential association with type D1-4 can be considered as a response when under heat-stress conditions, and that could help corals to cope with ocean warming.


Suggestion of a major gene for familial febrile convulsions mapping to 8q13-21.
Febrile convulsions affect 2 to 5% of all children under the age of 5 years. These convulsions probably have a variety of causes, but a genetic component has long been recognised. A large and remarkable family is described in which febrile convulsions appear to result from autosomal dominant inheritance at a single major locus. A gene for febrile convulsions was excluded from regions of previously mapped epilepsy genes and extension of exclusion mapping, using microsatellite markers, to the entire genome implied that a locus on chromosome 8q13-21 may be involved. Linkage analysis of markers on chromosome 8 gave a multipoint lod score of 3.40, maximised over different values of penetrance and phenocopy rate, for linkage between the gene for febrile convulsions and the region flanked by markers D8S553 and D8S279. This lod score was calculated assuming the disease has a penetrance of 60% and a phenocopy rate of 3%. Although there was no indication of linkage other than to markers on chromosome 8, linkage remains suggestive rather than significant because of the maximisation procedure applied. The support for linkage involving a major gene, as opposed to an alternative hypothesis of a complex inheritance pattern, relied upon the assumption of low penetrance.


Electrochemically driven biocatalysis of the oxygenase domain of neuronal nitric oxide synthase in indium tin oxide nanoparticles/polyvinyl alcohol nanocomposite.
Nitric oxide synthase (NOS) plays a critical role in a number of key physiological and pathological processes. Investigation of electron-transfer reactions in NOS would contribute to a better understanding of the nitric oxide (NO) synthesis mechanism. Herein, we describe an electrochemically driven catalytic strategy, using a nanocomposite that consisted of the oxygenase domain of neuronal NOS (D290nNOSoxy), indium tin oxide (ITO) nanoparticles and polyvinyl alcohol (PVA). Fast direct electron transfer between electrodes and D290nNOSoxy was observed with the heterogeneous electron transfer rate constant (ket) of 154.8 ± 0.1s(-1) at the scan rate of 5 Vs(-1). Moreover, the substrate N(ω)-hydroxy-L-arginine (NHA) was used to prove the concept of electrochemically driven biocatalysis of D290nNOSoxy. In the presence of the oxygen cosubstrate and tetrahydrobiopterin (BH4) cofactor, the addition of NHA caused the decreases of both oxidation current at +0.1 V and reduction current at potentials ranging from -0.149 V to -0.549 V vs Ag/AgCl. Thereafter, a series of control experiments such as in the absence of BH4 or D290nNOSoxy were performed. All the results demonstrated that D290nNOSoxy biocatalysis was successfully driven by electrodes in the presence of BH4 and oxygen. This novel bioelectronic system showed potential for further investigation of NOS and biosensor applications.


Folklore in children's odontology.
Popular customs with regard to tooth replacement are closely linked to magical ideas. The most important ones--such as throwing the tooth on the roof, giving it to mice or hiding it in different places--occur in places which are very far distant from one another, indicating that these customs originate from ancestral cultures.


Pathway and rate-limiting step of glyphosate degradation by Aspergillus oryzae A-F02.
Aspergillus oryzae A-F02, a glyphosate-degrading fungus, was isolated from an aeration tank in a pesticide factory. The pathway and rate-limiting step of glyphosate (GP) degradation were investigated through metabolite analysis. GP, aminomethylphosphonic acid (AMPA), and methylamine were detected in the fermentation liquid of A. oryzae A-F02, whereas sarcosine and glycine were not. The pathway of GP degradation in A. oryzae A-F02 was revealed: GP was first degraded into AMPA, which was then degraded into methylamine. Finally, methylamine was further degraded into other products. Investigating the effects of the exogenous addition of substrates and metabolites showed that the degradation of GP to AMPA is the rate-limiting step of GP degradation by A. oryzae A-F02. In addition, the accumulation of AMPA and methylamine did not cause feedback inhibition in GP degradation. Results showed that degrading GP to AMPA was a crucial step in the degradation of GP, which determines the degradation rate of GP by A. oryzae A-F02.


Structural and functional characterization of ScsC, a periplasmic thioredoxin-like protein from Salmonella enterica serovar Typhimurium.
The prototypical protein disulfide bond (Dsb) formation and protein refolding pathways in the bacterial periplasm involving Dsb proteins have been most comprehensively defined in Escherichia coli. However, genomic analysis has revealed several distinct Dsb-like systems in bacteria, including the pathogen Salmonella enterica serovar Typhimurium. This includes the scsABCD locus, which encodes a system that has been shown via genetic analysis to confer copper tolerance, but whose biochemical properties at the protein level are not defined. The aim of this study was to provide functional insights into the soluble ScsC protein through structural, biochemical, and genetic analyses. Here we describe the structural and biochemical characterization of ScsC, the soluble DsbA-like component of this system. Our crystal structure of ScsC reveals a similar overall fold to DsbA, although the topology of β-sheets and α-helices in the thioredoxin domains differ. The midpoint reduction potential of the CXXC active site in ScsC was determined to be -132 mV versus normal hydrogen electrode. The reactive site cysteine has a low pKa, typical of the nucleophilic cysteines found in DsbA-like proteins. Deletion of scsC from S. Typhimurium elicits sensitivity to copper (II) ions, suggesting a potential involvement for ScsC in disulfide folding under conditions of copper stress. ScsC is a novel disulfide oxidoreductase involved in protection against copper ion toxicity.


A single-domain antibody fragment in complex with RNase A: non-canonical loop structures and nanomolar affinity using two CDR loops.
Camelid serum contains a large fraction of functional heavy-chain antibodies - homodimers of heavy chains without light chains. The variable domains of these heavy-chain antibodies (VHH) have a long complementarity determining region 3 (CDR3) loop that compensates for the absence of the antigen-binding loops of the variable light chains (VL). In the case of the VHH fragment cAb-Lys3, part of the 24 amino acid long CDR3 loop protrudes from the antigen-binding surface and inserts into the active-site cleft of its antigen, rendering cAb-Lys3 a competitive enzyme inhibitor. A dromedary VHH with specificity for bovine RNase A, cAb-RN05, has a short CDR3 loop of 12 amino acids and is not a competitive enzyme inhibitor. The structure of the cAb-RN05-RNase A complex has been solved at 2.8 A. The VHH scaffold architecture is close to that of a human VH (variable heavy chain). The structure of the antigen-binding hypervariable 1 loop (H1) of both cAb-RN05 and cAb-Lys3 differ from the known canonical structures; in addition these H1 loops resemble each other. The CDR3 provides an antigen-binding surface and shields the face of the domain that interacts with VL in conventional antibodies. VHHs adopt the common immunoglobulin fold of variable domains, but the antigen-binding loops deviate from the predicted canonical structure. We define a new canonical structure for the H1 loop of immunoglobulins, with cAb-RN05 and cAb-Lys3 as reference structures. This new loop structure might also occur in human or mouse VH domains. Surprisingly, only two loops are involved in antigen recognition; the CDR2 does not participate. Nevertheless, the antigen binding occurs with nanomolar affinities because of a preferential usage of mainchain atoms for antigen interaction.


Double burden of underweight and overweight among Bangladeshi adults differs between men and women: evidence from a nationally representative survey.
To estimate the prevalence of underweight and overweight among Bangladeshi adults and to determine if the double burden of underweight and overweight differs by gender and other socio-economic characteristics of individuals. We used data from the Bangladesh Demographic and Health Survey 2011. Multinominal logistic regression was used to examine associations between the different nutritional statuses of individuals and related determinants. Interaction effect was checked between gender and various socio-economic factors. Nationwide, covering the whole of Bangladesh. Individuals aged >18 years (women, n 16 052; men, n 5090). Underweight was observed among 28·3 % of men and 24·4 % of women, whereas overweight was observed among 8·4 % of men and 16·9 % of women. The odds of being overweight were significantly lower among urban men (OR=0·46; 95 % CI 0·37, 0·57) compared with urban women, whereas the odds of being underweight were significantly higher among urban men (OR=1·33; 95 % CI 1·07, 1·64) compared with urban women. The odds of being overweight were lower among higher educated men (OR=0·48; 95 % CI 0·39, 0·58) and men of rich households (OR=0·45; 95 % CI 0·37, 0·54) compared with higher educated women and women of rich households, respectively. There are important gender differences in the prevalence of underweight and overweight among the adult population in Bangladesh. Women with higher education, in rich and urban households have higher chances of being overweight and lower chances of being underweight compared with their male counterparts.


The Epidemiology of HIV in Oman, 1984-2018: A Nationwide Study from the Middle East.
We used population-based data on all diagnosed people living with Human Immunodeficiency (HIV) reported to the National AIDS Programme in 1984-2018 to describe the HIV epidemiology in Oman. A total of 3060 Omanis were diagnosed with HIV from 1984 to 2018. The proportions of new infections attributed to sexual contact accounted for 56.3% (376/668) in 1984-1996 compared with 80.7% (630/780) in 2013-2018. Of 1417 patients with a documented CD4 count at the entry of care, 45.3% had a baseline CD4 count of <200 cells/mm3. Compared with heterosexuals, homosexuals had higher rates of advanced HIV disease [42.7% (388/908) vs 50.4% (136/270), respectively]. Rates of advanced disease and death within a year of HIV diagnosis rose consistently with age at diagnosis. Approximately half (48.8%) of the patients diagnosed in 1984-2018 had died by December 2018. The majority (85.6%; 572/668) of people who were diagnosed in 1984-1997 had died compared with 12.7% (99/780) of those diagnosed in 2013-2018. However, people died more recently had a higher proportion of death within a year of HIV diagnosis [74.7% (74/99) in 2013-2018 compared with 13.8% (79/572) in 1984-1996]. This study shows that the HIV epidemic in Oman is a low-prevalence one. Of concern, a large proportion of new HIV diagnoses continued to present late, which has resulted in a substantial increase in short-term mortality over the past 20 years. Nevertheless, we observed a remarkable decline in overall mortality over time, which may be explained by the improvement in the quality of HIV care in Oman.


Balancing the benefits of patient-clinician relationships with professional boundaries in Home-based Primary Care.
In the home care setting, boundaries may be difficult to identify when behavioral changes are made to accommodate the nature of care being delivered. In this secondary qualitative study, we examined how Home-based Primary Care (HBPC) clinicians understand role and relationship boundaries with patients and how these dynamics support patient care. The data set consisted of 14 semi-structured interviews with HBPC clinicians representing multiple disciplines and field observations of 6 HBPC team meetings. Using a directed approach to content analysis, we identified and described how HBPC clinicians worked to build relationships with patients, experienced challenges with emotional attachment, and negotiated boundaries in the patient-clinician relationship. Our findings illustrate how the home care setting is a site for which strong, therapeutic patient-clinician relationships can be developed while also highlighting the work that clinicians must do to balance addressing patient needs stemming from social isolation and adherence to their own professional boundaries.


Factors at play in faster progression for female pathological gamblers: an exploratory analysis.
Previous studies reported a faster progression for alcohol dependence and pathological gambling among females as compared with males. This phenomenon was called the "telescoping effect." By comparing female gamblers with male gamblers regarding gambling preferences and comorbidity, the authors explored potential risk factors for telescoping. A consecutive sample of Brazilian treatment-seeking pathological gamblers (DSM-IV criteria) was recruited. Genders were contrasted regarding comorbidity and gambling behavior, controlling for demographics, gambling severity, and previous access to mental health services. Seventy female gamblers and 70 male gamblers were interviewed. A greater proportion of women than men reported electronic bingo and video lottery terminals as their main type of gambling. Gambling was divided in 3 progressive stages: "social gambling," "intense gambling," and "problem gambling." Faster progression for female gamblers was confirmed; female gender and preference for electronic bingo and/or video lottery terminals were risk factors for telescoping throughout all stages. Female gamblers presented a higher comorbidity with depression, whereas male gamblers had higher rates of alcohol dependence. Nevertheless, comorbidity profiles were not related to gambling progression. Two factors could be at play for treatment-seeking female gamblers in Brazil: (1) a potential gender vulnerability and (2) a cultural environment yielding them access to a narrower range of gambling games that includes mainly the most addictive ones.


Collaborative Action of Surface Chemistry and Topography in the Regulation of Mesenchymal and Epithelial Markers and the Shape of Cancer Cells.
Malignant transformation is associated with enhancement of cell plasticity, which allows cancer cells to survive under different conditions by adapting to their microenvironment during growth and metastatic spread. Much effort has been devoted to understanding the molecular mechanisms of these processes. Although the importance of the extracellular matrix and of surface properties in these mechanisms is evident, the direct impact of distinct physical and chemical surfaces characteristics on cell fate remains unclear. Here, we have addressed this question using HT1080 fibrosarcoma cells as a model. To examine the relationship between surface topography, chemistry, and cell behavior, hydrophobic poly(butyl methacrylate-co-ethylene dimethacrylate) (BMA-EDMA) and hydrophilic poly(2-hydroxyethyl methacrylate-co-ethylene dimethacrylate) (HEMA-EDMA) surfaces with three different topographies (microporous, nanoporous, and nonporous) were generated. These surfaces were then modified by photoinitiated grafting of three different methacrylate monomers to create surface chemistry gradients of either negatively (AMPS) or positively (META) charged or zwitterionic (MDSA) functionalities. Our results show that AMPS promotes cell spreading, but that META abolishes cell growth. META and MDSA grafted on microporous BMA-EDMA produced superhydrophilic surfaces with high globularity and elasticity, which modified the cell phenotype by inhibiting cell spreading, followed by loss of mesenchymal characteristics and a reduction in protein levels of the mesenchymal markers N-cadherin, beta-catenin, p120 catenin, and also of the adaptor proteins vinculin and paxillin that are associated with adhesion and cancer cell invasion. The effect was strengthened along the gradient, suggesting that the density of the functional groups plays a role in this process. On the nanoporous surface, only MDSA grafting resulted in a significant increase in cell number, a reduction in N-cadherin expression, increased beta-catenin and p120 catenin levels, as well as the appearance of the epithelial marker E-cadherin. This indicates that the cancer cells have a high plasticity that is triggered by the collaborative effect of physical and chemical surface properties.


Regulatory T-Cell Therapy in Liver Transplantation and Chronic Liver Disease.
The constant exposure of the liver to gut derived foreign antigens has resulted in this organ attaining unique immunological characteristics, however it remains susceptible to immune mediated injury. Our understanding of this type of injury, in both the native and transplanted liver, has improved significantly in recent decades. This includes a greater awareness of the tolerance inducing CD4+ CD25+ CD127low T-cell lineage with the transcription factor FoxP3, known as regulatory T-Cells (Tregs). These cells comprise 5-10% of CD4+ T cells and are known to function as an immunological "braking" mechanism, thereby preventing immune mediated tissue damage. Therapies that aim to increase Treg frequency and function have proved beneficial in the setting of both autoimmune diseases and solid organ transplantations. The safety and efficacy of Treg therapy in liver disease is an area of intense research at present and has huge potential. Due to these cells possessing significant plasticity, and the potential for conversion towards a T-helper 1 (Th1) and 17 (Th17) subsets in the hepatic microenvironment, it is pre-requisite to modify the microenvironment to a Treg favourable atmosphere to maintain these cells' function. In addition, implementation of therapies that effectively increase Treg functional activity in the liver may result in the suppression of immune responses and will hinder those that destroy tumour cells. Thus, fine adjustment is crucial to achieve this immunological balance. This review will describe the hepatic microenvironment with relevance to Treg function, and the role these cells have in both native diseased and transplanted livers.


EDRF and the regulation of vascular tone.
Vascular endothelium is now known to produce a vasodilator agent, known as endothelium derived relaxing factor (EDRF) and recently shown to be nitric oxide. It acts locally on subjacent vascular smooth muscle by stimulating soluble guanylate cyclase to increase cyclic GMP levels and so reduce cytosolic free calcium. EDRF activity varies widely between different artery types and can be stimulated by various pharmacological agents and by flow. Studies in the intact vascular bed of the buffer-perfused rabbit ear show that EDRF activity is high in resistance vessels (particularly those of ca. 150 microns diameter), that EDRF is responsible for a 4th power relationship of diameter to flow in response to acute changes in flow (so limiting pressure gradients needed to increase flow rate), and that basal EDRF activity maintains geometric similarity of vessel diameters (implying that the spatial distribution of flow remains constant at different flow rates).


Early cerebral infarction following aneurysmal subarachnoid hemorrhage: frequency, risk factors, patterns, and prognosis.
Early cerebral infarction (ECI) following aneurysmal subarachnoid hemorrhage (aSAH) remains poorly understood. This study aims to determine the frequency and risk factors of this special episode, as well as to assess the relationship between its patterns and outcome. We retrospectively enrolled 243 patients who underwent aneurysm treatment within 60 hours of SAH. ECI was defined as one or more new hypodense abnormalities on computed tomography within 3 days after SAH, rather than lesions attributable to edema, retraction effect, and ventricular drain placement. Risk factors were tested by multivariate analysis. The infarct was classified by an established grading system (single or multiple, cortical or deep or combined). Poor outcome was defined as the Glasgow Outcome Score of severe disability or worse. Sixty-five patients (26.7%) had early infarction. Acute hydrocephalus (odds ratio [OR] 6.67; 95% confidence interval [CI] 1.59-27.95), admission plasma glucose level (OR 1.42 per mmol/L; 95% CI 1.16-1.73), and treatment modality (OR 16.27; 95% CI 4.05-65.28) were independent predictors of ECI. The pattern was single cortical in 19 patients (29.2%), single deep in 9 (13.8%), multiple cortical in 8 (12.3%), multiple deep in 14 (21.5%), and multiple combined in 15 (23.1%). ECI was associated with delayed cerebral infarction (DCI) (P = 0.002) and poor outcome (P < 0.001). Multiple combined infarction was related to poor outcome (P = 0.001). In summary, the occurrence of ECI, which is associated with surgical treatment, acute hydrocephalus and high admission plasma glucose, may potentially predict DCI and unfavorable outcome. Further studies are warranted to reveal the underlying mechanisms of this event and thereby minimize it.


[Paramagnetic properties of hepatic tissues and transplantable hepatomas].
Paramagnetic properties of frozen samples of normal, fetal and regenerating liver tissues and of a number transplantable hepatomas of different level of differentiation were studied. The results obtained indicate that ESR spectrum character and the concentration of paramagnetic centres in frozen samples of live tissues as well reflect the differentiation level, and the level of tissue functional activity. It is shown of hepatoma 22 taken as an example that tumour growth is accompanied with a change of the content of free radicals and some paramagnetic centres in the tumour and hosts liver. It can be considered as a manifestation of a biophysical shift which characterizes the tumour growth and includes the system effect of the tumour on the organism.


Evidence for a pH-dependent irreversible formation of a stable conformation of phenacyl-alpha-chymotrypsin.
A reinvestigation of the modification reactions of alpha-chymotrypsin with phenacyl bromide was carried out. Results conclusively demonstrate that the chemically and physically different modified enzymes prepared at pH 4 and at pH 7 both contain the phenacyl group at methionine-192 in the sulphonium salt form. Evidence to suppoort this conclusion derives from 13C nuclear-magnetic-resonance spectroscopic observations on [methylene-13C]phenacyl-enriched enzymes. More conclusively, the methionine-192-containing C-chain, derived by performic acid oxidative cleavage of radioactively-labelled enzyme prepared at pH 7, was shown to contain the phenacyl moiety and to undergo dealkylation by 2-mercaptoethanol with loss of this moiety. In addition, thermolytic cleavage of the high-pH enzyme results in fragmentation of the polypeptide chain in a fashion analogous to model reactions of phenacylmethionyl dipeptides and other methionine-192 sulphonium salts. A rationalization of the unusual nature of the high-pH phenacyl-modified enzyme based on the irreversible formation of stable conformation in which the phenacyl moiety is rigidly located in interior regions of the enzyme is presented and discussed.


Photobacterium swingsii sp. nov., isolated from marine organisms.
Six Gram-negative coccobacilli were isolated from Pacific oysters (Crassostrea gigas) from Mexico and haemolymph of spider crabs (Maja brachydactyla) from Spain. All of the isolates grew as small green colonies on thiosulphate-citrate-bile salts-sucrose (TCBS) agar and were facultatively anaerobic, oxidase-positive and sensitive to the vibriostatic agent O/129. Repetitive palindromic PCR analysis revealed a high degree of genomic homogeneity among the isolates. Several phenotypic traits differentiated the isolates from the type strains of species of the genus Photobacterium. DNA-DNA relatedness between two representative isolates and their closest phylogenetic neighbours by 16S rRNA gene sequence similarity, Photobacterium aplysiae CAIM 14(T) and Photobacterium frigidiphilum CAIM 20(T), was 44.01-53.85 %. We propose a novel species of the genus Photobacterium to accommodate the six isolates, with the name Photobacterium swingsii sp. nov. The type strain is CAIM 1393(T) (=CECT 7576(T)).