A novel Tembusu virus isolated from goslings in China form a new subgenotype 2.1.1.
Since 2010, several duck Tembusu viruses (DTMUVs) have been isolated from infected ducks in China, and these virus strains have undergone extensive variation over the years. Although the infection rate is high, the mortality rate is usually relatively low-~5%-30%; however, since fall 2019, an infectious disease similar to DTMUV infection but with a high mortality rate of ~50% in goslings has been prevalent in Anhui Province, China. The present study identified a new Tembusu virus, designated DTMUV/Goose/China/2019/AQ-19 (AQ-19), that is believed to be responsible for the noticeably high mortality in goslings. To investigate the genetic variation of this strain, its entire genome was sequenced and analysed for specific variations, and goslings and mice were challenged with the isolated virus to investigate its pathogenicity. The AQ-19 genome shared only 94.3%-96.9% and 90.9% nucleotide identity with other Chinese and Malaysian DTMUVs, respectively; however, AQ-19 has high homology with Thailand DTMUVs (97.2%-98.1% nucleotide identity). Phylogenetic analysis of the E gene revealed that AQ-19 and most of Thailand DTMUVs form a branch separate from any of the previously reported DTMUV strains in China. After the challenge, some goslings and mice showed typical clinical signs of DTMUV, particularly severe neurological dysfunction. AQ-19 has high virulence in goslings and mice, resulting in 60% and 70% mortality through intramuscular and intracerebral routes, respectively. Pathological examination revealed severe histological lesions in the brain and liver of the infected goslings and mice. Taken together, these results demonstrated the emergence of a novel Tembusu virus with high virulence circulating in goslings in China for the first time, and our findings highlight the high genetic diversity of DTMUVs in China. Further study of the pathogenicity and host range of this novel Tembusu virus is particularly important.


Frequent occurrence of respiratory symptoms in children is associated with exposure to air pollution, land use types, and parental mental health in the Greater Taipei area.
Although studies have investigated the individual effects of air pollution, land use types, and parental mental health on children's respiratory health, few studies have examined the effects of these risk factors simultaneously in children aged <2 years. We investigated the effects of exposure to air pollution, land use types surrounding residences, and parental mental health on the frequent occurrence of respiratory symptoms in children aged <2 years in the Greater Taipei area. Participants were recruited from an ongoing Taiwanese birth cohort study. We analyzed the data of the participants who had been recruited from January 2011 to April 2014 and had responded to the follow-up questionnaires at 6, 12, and 24 months. Self-administered questionnaires were used to collect participants' sociodemographic background and health, such as respiratory symptoms, and parental mental health. Pre- and postnatal pollution levels were estimated using the spatial interpolation technique (ordinary kriging) at children's residential addresses. Land use types surrounding participants' homes were evaluated by performing buffer analysis. Multiple logistic regression analyses were conducted to examine the effects of risk factors on the frequent occurrence of child respiratory symptoms in children aged 6, 12, and 24 months. We included 228, 360, and 441 children aged 6, 12, and 24 months, respectively. Our results indicated that postnatal exposure to PM2.5 and O3 was positively associated with children's respiratory symptoms. Traffic-related land-use types, sports facilities, and commercial land surrounding homes exerted adverse effects on children's respiratory symptoms, whereas the presence of schools in the neighborhood was beneficial. Parental mental health was also associated with children's respiratory symptoms. Postnatal exposure to air pollution and land use types surrounding residences were associated with respiratory health in children aged <2 years. The residential environment is a critical factor affecting children's respiratory health of children aged <2 years.


Are we teaching sufficient anatomy at medical school? The opinions of newly qualified doctors.
In recent decades wide-ranging changes have occurred in medical school curricula. Time spent studying gross anatomy has declined amidst controversy as to how, what, and when teaching is best delivered. This reduced emphasis has led to concerns amongst clinicians that a new generation of doctors are leaving medical school with insufficient anatomical knowledge. Previous studies have established that medical students value their anatomy teaching during medical school. None have sought to establish views on the sufficiency of this teaching. We investigate the opinions of newly qualified doctors at a UK medical school and relate these opinions to career intentions and academic performance in the setting of a traditional dissection and prosection-based course. Overall nearly half of respondents believe they received insufficient anatomy teaching. A substantial proportion called for the integration of anatomy teaching throughout the medical school course. Trainees intent on pursuing a surgical career were more likely to believe anatomy teaching was insufficient than those pursuing a nonsurgical career; however, overall there was no statistical difference in relation to the mean for any individual career group. This study adds to the current debates in anatomical sciences education, indicating that overall, regardless of career intentions, new doctors perceive the need for greater emphasis on anatomical teaching.


Expression and possible function of IL-2 and IL-15 receptors on human uveal melanoma cells.
Interleukin-2 (IL-2) and IL-15 receptors have been detected on some murine neoplasms. Accordingly, the expression of these receptors on human uveal melanoma cell lines was examined, and the effect of exogenous IL-2 and -15 on melanoma cell proliferation, susceptibility to natural killer (NK) cell-mediated cytolysis, and sensitivity to apoptosis were assessed. Nine human uveal melanoma cell lines and three cell lines from uveal melanoma metastases were tested by flow cytometry for the expression of human IL-2R and -15Ralpha. Melanoma cells were cultured, with or without recombinant human IL-2 or -15, cell proliferation was determined by tritiated thymidine incorporation, and IL-2 and -15 receptor expression was assessed by flow cytometry. The effect of these cytokines on NK activity was evaluated with a standard (51)Cr-release assay. All the melanoma cell lines expressed IL-2R and -15R. IL-2 induced a three- to eightfold upregulation of IL-2R expression in all the melanoma cell lines. Although IL-2 did not affect the proliferation of six of the seven uveal melanoma cell lines, it induced a 32% and 57% increase in the proliferation of both metastatic cell lines. IL-15 induced proliferation on all tested cell lines (4%-68%). Both IL-2 and -15 reduced melanoma cell sensitivity to NK-cell-mediated cytolysis and cisplatin-induced apoptosis. The results suggest that IL-2 and -15 elaborated by tumor-infiltrating lymphocytes and macrophages may affect the malignant behavior of human uveal melanoma by stimulating proliferation and reducing uveal melanoma cell susceptibility to NK-cell-mediated cytolysis and cisplatin-induced apoptosis.


Hydrogen-bonded complexes of lumichrome.
Hydrogen bonds were shown to play an important role in the lumichrome photophysics and photochemistry both in solutions and in the solid state. In solutions, lumichrome can form hydrogen-bonded complexes with a variety of molecules, such as acetic acid or methanol, as supported by spectral and equilibrium studies. Photoexcitation of some hydrogen-bonded complexes, having appropriate configuration, as in the case of acetic acid, may lead to excited-state proton transfer, resulting in formation of the isoalloxazinic structure, detectable by its characteristic emission, distinct from that of the intrinsically alloxazinic lumichrome. Theoretical calculations confirmed the role of the hydrogen-bonded complexes, yielding several stable eight-membered cyclic structures of such complexes characterized by spectral changes similar to those observed experimentally. Hydrogen bonds play an essential role in the formation of the lumichrome crystal structure, as follows from the X-ray diffraction results. Interestingly, the crystals studied included molecules of methanol used as solvent in crystal growth. The emission studies of polycrystalline samples, similar to the processes occurring in solutions, point to the importance of hydrogen-bonding interactions in crystal packing allowed by the symmetry of the hydrogen-bonded dimers.


Maturity assessment of software-driven medical technologies: a quantitative score derived from a quality standard for the research phase.
Quantitatively assessing the level of readiness of medical technology improves its chance of successfully transfer from research to industry but remains a challenge. As many innovative medical devices are associated with or incorporate software, this article presents a methodology for evaluating the software maturity of a "Software-driven Medical Technology" (SdMT) during the research phase. A technological maturity model is developed by methodologically extracting relevant terms from the ISO/IEC 62304 standard, the main industry standard for medical device software, and results in a list of required software engineering artifacts. This list and the relative weight of the artifacts are used to establish a software maturity score for SdMT and the corresponding assessment questionnaire. The consistency of the model is demonstrated by analyzing the obtained score system relatively with the standard. The maturity score of a SdMT can be assessed during the research phase and depends on the number and importance of the artifacts already present at the time of evaluation.Clinical relevance- The proposed quantitative maturity score can help the medical technology innovation actors (clinicians, researchers and industrials) to better identify, improve and fasten the readiness of technology for clinical investigation and technology transfer.


Climate change and eutrophication induced shifts in northern summer plankton communities.
Marine ecosystems are undergoing substantial changes due to human-induced pressures. Analysis of long-term data series is a valuable tool for understanding naturally and anthropogenically induced changes in plankton communities. In the present study, seasonal monitoring data were collected in three sub-basins of the northern Baltic Sea between 1979 and 2011 and statistically analysed for trends and interactions between surface water hydrography, inorganic nutrient concentrations and phyto- and zooplankton community composition. The most conspicuous hydrographic change was a significant increase in late summer surface water temperatures over the study period. In addition, salinity decreased and dissolved inorganic nutrient concentrations increased in some basins. Based on redundancy analysis (RDA), warming was the key environmental factor explaining the observed changes in plankton communities: the general increase in total phytoplankton biomass, Cyanophyceae, Prymnesiophyceae and Chrysophyceae, and decrease in Cryptophyceae throughout the study area, as well as increase in rotifers and decrease in total zooplankton, cladoceran and copepod abundances in some basins. We conclude that the plankton communities in the Baltic Sea have shifted towards a food web structure with smaller sized organisms, leading to decreased energy available for grazing zooplankton and planktivorous fish. The shift is most probably due to complex interactions between warming, eutrophication and increased top-down pressure due to overexploitation of resources, and the resulting trophic cascades.


Three-Dimensional Printing: Custom-Made Implants for Craniomaxillofacial Reconstructive Surgery.
Craniomaxillofacial reconstructive surgery is a challenging field. First it aims to restore primary functions and second to preserve craniofacial anatomical features like symmetry and harmony. Three-dimensional (3D) printed biomodels have been widely adopted in medical fields by providing tactile feedback and a superior appreciation of visuospatial relationship between anatomical structures. Craniomaxillofacial reconstructive surgery was one of the first areas to implement 3D printing technology in their practice. Biomodeling has been used in craniofacial reconstruction of traumatic injuries, congenital disorders, tumor removal, iatrogenic injuries (e.g., decompressive craniectomies), orthognathic surgery, and implantology. 3D printing has proven to improve and enable an optimization of preoperative planning, develop intraoperative guidance tools, reduce operative time, and significantly improve the biofunctional and the aesthetic outcome. This technology has also shown great potential in enriching the teaching of medical students and surgical residents. The aim of this review is to present the current status of 3D printing technology and its practical and innovative applications, specifically in craniomaxillofacial reconstructive surgery, illustrated with two clinical cases where the 3D printing technology was successfully used.


Drug administration via enteral feeding tubes in intensive therapy - terra incognita?
The use of enteral feeding tubes has become more frequent, both in hospital settings and in home care. The feeding tubes serve not only to deliver nutrients, but also as a route for medication provision. Nonetheless, the pharmaceutical, legal and technical implications of medication delivery via enteral feeding tubes are not widely understood by doctors and nurses. Not only is the type of medication relevant, but also the type of feeding tube. Crushing tablets may have detrimental effects for a patient and a staff member too. Administering a drug via enteral feeding tubes usually falls outside the terms of the licence (off-label), so burdening medical staff with the entire responsibility for potential adverse reactions.


Microplate spectroscopic methods for determination of the organophosphate soman.
Two microplate spectroscopic methods for determination of organophosphates, based on inhibition of acetylcholinesterase activity, have been elaborated and evaluated for determination of the chemical weapon agent soman. The principal difference between the methods is that one measures reaction substrate concentration (elaborated from Hestrin), while the other measures reaction product (elaborated from Ellman). The linear ranges of the two methods were found to be similar. Although the limit of quantification was lower for the Ellman method (110 pM), the sensitivity coefficient was in favor of the Hestrin method (1.55-fold higher). The effects of the main soman hydrolysis products were consistent for the two methods: both methylphosphonic acid and pinacolyl methylphosphonic acid did not inhibit acetylcholinesterase activity. The main components of decontaminating solutions showed differential effects: while monoethanolamine had no influence upon results obtained by either method, hydrogen peroxide interfered with the Ellman method at far lower concentrations than with the Hestrin method. In practical applications involving samples containing hydrogen peroxide, the method based on Hestrin should be regarded as much more specific for OP determination than the Ellman method.


Clinical outcomes among persons with pulmonary tuberculosis caused by Mycobacterium tuberculosis isolates with phenotypic heterogeneity in results of drug-susceptibility tests.
Patients with multidrug-resistant (MDR) tuberculosis may have phenotypic heterogeneity in results of drug-susceptibility tests (DSTs). However, the impact of this on clinical outcomes among patients treated for MDR tuberculosis is unknown. Phenotypic DST heterogeneity was defined as presence of at least 1 Mycobacterium tuberculosis isolate susceptible to rifampicin and isoniazid recovered <3 months after MDR tuberculosis treatment initiation from a patient with previous documented tuberculosis due to M. tuberculosis resistant to at least rifampicin and isoniazid. The primary outcome was defined as good (ie, cure or treatment completion) or poor (ie, treatment failure, treatment default, or death). A secondary outcome was time to culture conversion. Cox proportional hazard models were used to determine the association between phenotypic DST heterogeneity and outcomes. Phenotypic DST heterogeneity was identified in 33 of 475 patients (7%) with MDR tuberculosis. Poor outcome occurred in 126 patients (28%). Overall, patients with MDR tuberculosis who had phenotypic DST heterogeneity were at greater risk of poor outcome than those with MDR tuberculosis but no phenotypic DST heterogeneity (adjusted hazard ratio [aHR], 2.1; 95% confidence interval [CI], 1.2-3.6). Among HIV-infected patients with MDR tuberculosis, the adjusted hazard for a poor outcome for those with phenotypic DST heterogeneity was 2.4 (95% CI, 1.3-4.2) times that for those without phenotypic DST heterogeneity, whereas among HIV-negative patients with MDR tuberculosis, the adjusted hazard for those with phenotypic DST heterogeneity was 1.5 (95% CI, .5-4.3) times that for those without phenotypic DST heterogeneity. HIV-infected patients with MDR tuberculosis with phenotypic DST heterogeneity also had a longer time to culture conversion than with HIV-infected patients with MDR tuberculosis without phenotypic DST heterogeneity (aHR, 2.9; 95% CI, 1.4-6.0). Phenotypic DST heterogeneity among persons with HIV infection who are being treated for MDR tuberculosis is associated with poor outcomes and longer times to culture conversion.


[Predicting the outcome of nerve suture].
The late results of nerve suture were studied in 398 patients with injury of one or more nerves and coding cards containing formalized data of the primary examination of the patients and the operative data were filled in. After mathematical processing of these data on an electronic computer (CBM-6000) using the linear regression method, 4 formulas were obtained from which a physician practicing in any field of medicine can prognosticate in 5-7 min the degree of restoration of the limb function and the working rehabilitation of individuals who had suffered injury of the nerves.


Postmarketing surveillance of the safety of cimetidine: mortality during second, third, and fourth years of follow up.
A previous report analysed the pattern of mortality during the first year of follow up among 9928 patients taking cimetidine who were recruited to a postmarketing drug surveillance study in Glasgow, Nottingham, Oxford, and Portsmouth. A further analysis has now been conducted extending the period of follow up to four years. The 12 month report noted that cimetidine was being given, knowingly or unknowingly, in the late stages of many diseases and also to counter the adverse gastric effects of other drugs used in the treatment of serious disorders. This finding was underlined by a steady fall in the excess death rate among cimetidine users with increasing length of follow up, such that by the fourth year the pattern of observed deaths was not much different from that expected on the basis of national rates. Some excess of observed over expected deaths from gastric cancer, lung cancer, and urinary disorders was still apparent after four years of follow up, but there was no evidence that cimetidine was responsible. Indeed, no fatal disorder emerged as being associated with cimetidine during the follow up period. Deaths from the complications of disease related to gastric acid occurred in only 38 of the 9928 subjects over the four years. These findings provide further evidence of the safety of cimetidine.


Double-blind placebo-controlled study of concurrent administration of albendazole and praziquantel in schoolchildren with schistosomiasis and geohelminths.
A double-blind placebo-controlled study of the concurrent administration of albendazole and praziquantel was conducted in>1500 children with high prevalences of geohelminths and schistosomiasis. The study sites were in China and the Philippines, including 2 strains of Schistosoma japonicum, and 2 different regions of Kenya, 1 each with endemic Schistosoma mansoni or Schistosoma haematobium. Neither medication affected the cure rate of the other. There was no difference between the side effect rate from albendazole or the double placebo. Praziquantel-treated children had more nausea, abdominal pain, and headache but these side effects were statistically more common in children with schistosomiasis, suggesting a strong influence of dying parasites. The subjects were followed for 6 months for changes in infection status, growth parameters, hemoglobin, and schistosomiasis morbidity. In all 4 sites, a significant 6-month increase in serum hemoglobin was observed in children who received praziquantel, strongly supporting population-based mass treatment.


Differential effects of Wilms tumor WT1 splice variants on the insulin receptor promoter.
The Wilms tumor gene WT1 has been implicated in the early development of the kidney. Mutations in WT1 are found in a small fraction of Wilms tumor, a pediatric nephroblastoma, and Denys-Drash syndrome, characterized by genitourinary abnormalities. The WT1 gene product functions as a transcriptional repressor of growth factor-related genes. The kidney is one of the major sites of insulin action in vivo and expresses high levels of insulin receptors (IR). IR expression has been detected during early embryogenesis, suggesting that it may play a role in development. We investigated whether two WT1 splice variants lacking or including a three-amino-acid (KTS) insertion between the third and fourth zinc finger in the DNA-binding domain could repress the IR promoter in vitro. We show that the +KTS variant effectively represses promoter activity under all conditions tested but the -KTS variant was only able to repress in the presence of cotransfected C/EBP beta or a dominant-negative p53 mutation. Deletional mapping indicated that distinct regions of the IR promoter mediated the effects of the two isoforms and DNaseI footprint analysis identified potential WT1 binding sites within these regions.


Individual differences in functional brain connectivity predict temporal discounting preference in the transition to adolescence.
The transition from childhood to adolescence is marked by distinct changes in behavior, including how one values waiting for a large reward compared to receiving an immediate, yet smaller, reward. While previous research has emphasized the relationship between this preference and age, it is also proposed that this behavior is related to circuitry between valuation and cognitive control systems. In this study, we examined how age and intrinsic functional connectivity strength within and between these neural systems relate to changes in discounting behavior across the transition into adolescence. We used mixed-effects modeling and linear regression to assess the contributions of age and connectivity strength in predicting discounting behavior. First, we identified relevant connections in a longitudinal sample of 64 individuals who completed MRI scans and behavioral assessments 2-3 times across ages 7-15 years (137 scans). We then repeated the analysis in a separate, cross-sectional, sample of 84 individuals (7-13 years). Both samples showed an age-related increase in preference for waiting for larger rewards. Connectivity strength within and between valuation and cognitive control systems accounted for further variance not explained by age. These results suggest that individual differences in functionalbrain organization can account for behavioral changes typically associated with age.


Emerging Roles of Growth Differentiation Factor 15 in Immunoregulation and Pathogenesis.
Growth differentiation factor 15 (GDF-15) is a cytokine that is widely used as a biomarker for the severity of diverse disease states. It also has been shown to play a protective role after tissue injury and to promote a negative energy balance during obesity and diabetes. In addition to its metabolic effects, GDF-15 also regulates the host's immune responses to infectious and noninfectious diseases. GDF-15 can suppress a type 1 and, in contrast, promote a type 2 inflammatory response. In this brief review, we discuss how GDF-15 affects the effector function and recruitment of immune cells, the pathways that induce its expression, and the diverse mechanisms by which it is regulated during inflammation and infection. We further highlight outstanding questions that should be the focus of future investigations in this emerging field.


Structural insights into the assembly and mechanism of mpox virus DNA polymerase complex F8-A22-E4-H5.
The DNA replication of mpox virus is performed by the viral polymerase F8 and also requires other viral factors, including processivity factor A22, uracil DNA glycosylase E4, and phosphoprotein H5. However, the molecular roles of these viral factors remain unclear. Here, we characterize the structures of F8-A22-E4 and F8-A22-E4-H5 complexes in the presence of different primer-template DNA substrates. E4 is located upstream of F8 on the template single-stranded DNA (ssDNA) and is catalytically active, highlighting a functional coupling between DNA base-excision repair and DNA synthesis. Moreover, H5, in the form of tetramer, binds to the double-stranded DNA (dsDNA) region downstream of F8 in a similar position as PCNA (proliferating cell nuclear antigen) does in eukaryotic polymerase complexes. Omission of H5 or disruption of its DNA interaction showed a reduced synthesis of full-length DNA products. These structures provide snapshots for the working cycle of the polymerase and generate insights into the mechanisms of these essential factors in viral DNA replication.


Fish Consumption and Incidence of Diabetes: meta-analysis of data from 438,000 individuals in 12 independent prospective cohorts with an average 11-year follow-up.
Ecological data suggest an inverse correlation between fish consumption and diabetes prevalence. However, epidemiological data on fish intake and diabetes incidence are controversial and inconclusive. Therefore, we aimed to assess the literature and determine the association between fish consumption and diabetes risk quantitatively. Prospective cohort studies published through August 2011 in peer-reviewed journals indexed in PubMed were selected. Additional information was retrieved through Google or a hand search of the references from relevant articles. The weighted relative risk (RR) and corresponding 95% CI for incident diabetes was estimated using random-effects models. A database was derived from nine eligible studies (12 independent cohorts), including 438,214 individuals with an average 11.4-year follow-up. Compared with those who never consumed fish or ate fish less than once per month, the pooled RR of incident diabetes was 0.99 (95% CI 0.85-1.16) for individuals who ate fish five or more times per week (P(trend) = 0.80). Similar results were found for long-chain n-3 polyunsaturated fatty acid intake. Study location was an effect modifier. An inverse association between fish intake and diabetes incidence was found by combining studies conducted in Eastern but not Western countries. Accumulated evidence generated from this meta-analysis does not support an overall inverse association of fish or fish oil intake with incidence of diabetes. The null association was modified by study location (Eastern vs. Western countries), which may reflect the possible difference between Eastern and Western dietary patterns. Further studies are warranted.


Raised serum creatine phosphokinase activity in ankylosing spondylitis.
Serum enzyme studies were made on 43 (37 male, 6 female) consecutive patients with ankylosing spondylitis. Serum creatine phosphokinase (CPK) activity was raised above 55 IU/l in 24 (65%) of 37 male patients (range 29-165 IU/l, mean 68) as compared with 2 (4%) out of 47 male controls (range 14-85 IU/l, mean 33: P less than 0-001); levels were greater than 35 IU/l in six (100%) out of six female patients (range 39-100 IU/l, mean 56) as against one (3%) of 35 female controls (range 3-106 IU/l, mean 16; P less than 0-001). The recognized pitfalls in interpreting CPK activity were avoided. In all of sixteen randomly selected patients isoenzyme studies confirmed that muscle is the source of the enzyme. There was a significant correlation between CPK activity and both spinal flexion and the reciprocal of finger-to-floor distance (P less than 0-5 in each case).


Multiple cellular phenotypes in an insertional mutation in mouse affecting bone development.
The 6093 line of transgenic mice exhibits altered bone development as a result of an insertional mutation by the transgene. Female transgenic mice show a marked kyphosis as early as 2 weeks of age. Vertebrae from female mice have lower total bone area and mineral content than age-matched, gender-matched controls, although the bone mineral density is not changed. The femur and tibia exhibit the opposite effect-increased bone area and mineral content. Fluorescent bone label experiments indicated an increased rate of bone mineral deposition in the femur during the early postnatal growth period, and bone marrow from femurs of 6093 females had increased numbers of fibroblast colony-forming units. Transgenic females also are obese and have altered thymocyte development, suggesting that the insertional mutation affects multiple cell populations. We hypothesize that these phenotypes arise as a result of an alteration in the function or developmental potential of a stromal cell or mesenchymal stem cell.


Linear normalization of MR brain images in pediatric patients with periventricular leukomalacia.
The feasibility of linear normalization of child brain images with structural abnormalities due to periventricular leukomalacia (PVL) was assessed in terms of success rate and accuracy of the normalization algorithm. Ten T1-weighted brain images from healthy adult subject and 51 from children (4-11 years of age) were linearly transformed to achieve spatial registration with the standard MNI brain template. Twelve of the child brain images were radiologically normal, 22 showed PVL and 17 showed PVL with additional enlargement of the lateral ventricles. The effects of simple modifications to the normalization process were evaluated: changing the initial orientation and zoom parameters, masking non-brain areas, smoothing the images and using a pediatric template instead of the MNI template. Normalization failure was reduced by changing the initial zoom parameters and by removing background noise. The overall performance of the normalization algorithm was only improved when background noise was removed from the images. The results show that linear normalization of PVL affected brain images is feasible.


Ethnic variation in the correlation between fasting glucose concentration and glycated hemoglobin (HbA1c).
We aimed to determine the relationship between fasting serum glucose (FSG) concentration and glycated hemoglobin-A1c (HbA1c) in the 3 ethnicities in Singapore after adjustment for demographic and therapeutic variables. Fasting serum glucose (FSG), HbA1c, and serum creatinine levels were simultaneously sampled from 575 patients with diabetes (389 Chinese, 97 Indians, 89 Malays) in this cross-sectional study between January and May 2008, and the results were subjected to multivariate linear regression analysis. We found a significant interaction between FSG and ethnicity on HbA1c. The correlation between FSG and HbA1c among Chinese subjects was 0.25 (95% confidence interval [CI]:0.2-0.3) relative to the Malays (0.38, 95% CI: 0.30-0.45) after adjustment for age; gender; serum creatinine concentrations; body mass index (BMI); duration of diabetes; use of sulfonylureas, metformin, and insulin; and hemoglobin (Hb) and red cell indices (P = .005). Hence, for a given FSG, the predicted HbA1c will be higher in Malays compared to Chinese subjects. We did not observe a statistically significant difference between Indians and Malays with respect to the correlation between FSG and HbA1c. We showed a higher correlation between HbA1c and FSG in Malay subjects relative to the Chinese in this cohort. The ethnic variation in the HbA1c-FSG relationship may be related to differences in percentage contribution by the FSG to overall HbA1c among ethnic groups. Future studies using continuous glucose monitoring (CGM) to elucidate the relative contributions by FSG and postprandial glucose (PPG) to the daily blood glucose profile and the overall HbA1c by ethnicity are required.


The genetically modified polysialylated form of neural cell adhesion molecule-positive cells for potential treatment of X-linked adrenoleukodystrophy.
Cell transplantation of myelin-producing exogenous cells is being extensively explored as a means of remyelinating axons in X-linked adrenoleukodystrophy. We determined whether 3,3',5-Triiodo-L-thyronine (T3) overexpresses the ABCD2 gene in the polysialylated (PSA) form of neural cell adhesion molecule (NCAM)-positive cells and promotes cell proliferation and favors oligodendrocyte lineage differentiation. PSA-NCAM+ cells from newborn Sprague-Dawley rats were grown for five days on uncoated dishes in defined medium with or without supplementation of basic fibroblast growth factor (bFGF) and/or T3. Then, PSA-NCAM+ spheres were prepared in single cells and transferred to polyornithine/fibronectin-coated glass coverslips for five days to determine the fate of the cells according to the supplementation of these molecules. T3 responsiveness of ABCD2 was analyzed using real-time quantitative polymerase chain reaction, the growth and fate of cells were determined using 5-bromo-2-deoxyuridine incorporation and immunocytochemistry, respectively. Results demonstrated that T3 induces overexpression of the ABCD2 gene in PSA-NCAM+ cells, and can enhance PSA-NCAM+ cell growth in the presence of bFGF, favoring an oligodendrocyte fate. These results may provide new insights into investigation of PSA-NCAM+ cells for therapeutic application to X-linked adrenoleukodystrophy.


An immunological study of Lyme disease.
84 patients in the second or third stages of serologically diagnosed Lyme disease suffering from different forms of central as well as peripheral nervous system involvement were tested for the participation of autoimmune mechanisms. Cell hypersensitivity to the encephalitogenic basic protein and to antigens from peripheral myelin was evaluated on an Opton cytopherometer according to the effect of the macrophage slowing factor--liberated during short-term incubation of sensibilized lymphocytes by the respective antigen--on the mobility of tannin-treated sheep red blood cells in an electric field. Judged by the presence of autoimmune reactions. Lyme disease has come to occupy a prominent position compared with previously examined other types of neuroinfections. An explanation can be sought in disordered immunoregulating mechanisms, even though the changes in total and active T lymphocytes were not statistically significant. Data on immunological changes are relevant for the indication of immunomodulating therapy as a suitable complement to antibiotic treatment.


Molecular adaptability of nucleoside diphosphate kinase b from trypanosomatid parasites: stability, oligomerization and structural determinants of nucleotide binding.
Nucleoside diphosphate kinases play a crucial role in the purine-salvage pathway of trypanosomatid protozoa and have been found in the secretome of Leishmania sp., suggesting a function related to host-cell integrity for the benefit of the parasite. Due to their importance for housekeeping functions in the parasite and by prolonging the life of host cells in infection, they become an attractive target for drug discovery and design. In this work, we describe the first structural characterization of nucleoside diphosphate kinases b from trypanosomatid parasites (tNDKbs) providing insights into their oligomerization, stability and structural determinants for nucleotide binding. Crystallographic studies of LmNDKb when complexed with phosphate, AMP and ADP showed that the crucial hydrogen-bonding residues involved in the nucleotide interaction are fully conserved in tNDKbs. Depending on the nature of the ligand, the nucleotide-binding pocket undergoes conformational changes, which leads to different cavity volumes. SAXS experiments showed that tNDKbs, like other eukaryotic NDKs, form a hexamer in solution and their oligomeric state does not rely on the presence of nucleotides or mimetics. Fluorescence-based thermal-shift assays demonstrated slightly higher stability of tNDKbs compared to human NDKb (HsNDKb), which is in agreement with the fact that tNDKbs are secreted and subjected to variations of temperature in the host cells during infection and disease development. Moreover, tNDKbs were stabilized upon nucleotide binding, whereas HsNDKb was not influenced. Contrasts on the surface electrostatic potential around the nucleotide-binding pocket might be a determinant for nucleotide affinity and protein stability differentiation. All these together demonstrated the molecular adaptation of parasite NDKbs in order to exert their biological functions intra-parasite and when secreted by regulating ATP levels of host cells.


Spatially and spectrally resolved imaging of modal content in large-mode-area fibers.
A new measurement technique, capable of quantifying the number and type of modes propagating in large-mode-area fibers is both proposed and demonstrated. The measurement is based on both spatially and spectrally resolving the image of the output of the fiber under test. The measurement provides high quality images of the modes that can be used to identify the mode order, while at the same time returning the power levels of the higher-order modes relative to the fundamental mode. Alternatively the data can be used to provide statistics on the level of beam pointing instability and mode shape changes due to random uncontrolled fluctuations of the phases between the coherent modes propagating in the fiber. An added advantage of the measurement is that is requires no prior detailed knowledge of the fiber properties in order to identify the modes and quantify their relative power levels. Because of the coherent nature of the measurement, it is far more sensitive to changes in beam properties due to the mode content in the beam than is the more traditional M(2) measurement for characterizing beam quality. We refer to the measurement as Spatially and Spectrally resolved imaging of mode content in fibers, or more simply as S(2) imaging.


Maternal state of mind regarding attachment, maternal depression and children's family drawings in the early school years.
The current study explored how children's family drawings in the early school years might be related to their exposure to recurrent episodes of maternal depression. We also examined prospectively relations among maternal state of mind regarding attachment derived from the Adult Attachment Interview and earlier mother-child attachment from the Strange Situation Procedure (both measured when the child was 12-15 months old) and later family drawings. Seventy-five mothers were assessed for symptoms of depression periodically between birth and child age 6-8 years. At this age, children completed a family drawing rated using an attachment-based scoring system. Both mothers' state of mind regarding attachment and their overall depression were modestly, but significantly, correlated with a global rating of the child's drawings. When both predictors were considered together, however, neither was significant, reflecting collinearity between the two variables. In this study, women with a non-autonomous state of mind regarding attachment were significantly more likely to experience recurrent depression. The earlier classification of the child's attachment to the mother from the Strange Situation Procedure was not related to the family drawing.


Ezrin is required for efficient Rap1-induced cell spreading.
The Rap family of small GTPases regulate the adhesion of cells to extracellular matrices. Several Rap-binding proteins have been shown to function as effectors that mediate Rap-induced adhesion. However, little is known regarding the relationships between these effectors, or about other proteins that are downstream of or act in parallel to the effectors. To establish whether an array of effectors was required for Rap-induced cell adhesion and spreading, and to find new components involved in Rap-signal transduction, we performed a small-scale siRNA screen in A549 lung epithelial cells. Of the Rap effectors tested, only Radil blocked Rap-induced spreading. Additionally, we identified a novel role for Ezrin downstream of Rap1. Ezrin was necessary for Rap-induced cell spreading, but not Rap-induced cell adhesion or basal adhesion processes. Furthermore, Ezrin depletion inhibited Rap-induced cell spreading in several cell lines, including primary human umbilical vein endothelial cells. Interestingly, Radixin and Moesin, two proteins with high homology to Ezrin, are not required for Rap-induced cell spreading and cannot compensate for loss of Ezrin to rescue Rap-induced cell spreading. Here, we present a novel function for Ezrin in Rap1-induced cell spreading and evidence of a non-redundant role of an ERM family member.


[Relative abundance, population structure, habitat preferences and activity patterns of Tapirus bairdii (Perissodactyla: Tapiridae), in Chimalapas forest, Oaxaca, Mexico].
Baird's tapir (Tapirus bairdii) is endangered primarily because of habitat loss and fragmentation, and overhunting throughout its distribution range. One of the priority land areas for the conservation of this species is the Northern part of its range in the Chimalapas forest, Oaxaca. The aim of this research was to determine the relative abundance, population struc- ture, habitat preferences and activity patterns of Baird's tapir (Tapirus bairdii) in the Chimalapas forest, Oaxaca, Mexico, through the non-invasive technique of camera-trap sampling. A total of five sampling sessions were undertaken among 2009-2013, and used a total of 30 camera-traps in each period. The determinant factor of the sampling design was the hunting between two study areas. A total sampling effort of 9000 trap-days allowed to estimate an index of relative abundance (IRA) of 6.77 tapir photographs/1,000 trap-days (n = 61). IRA varied significantly between sampling stations (Mann-Whitney, p < 0.01). The frequency of Baird's tapir photos was higher in the dry season in tropical rain forest without hunting (χ2, p < 0.5). In the rainy season, the tropical rain forest and secondary vegetation habitats showed higher photo frequency than expected from random (χ2, p < 0.5). Considering population structure, a 95.08% of adult animals was obtained in photographic records (n = 58). Three types of activity pattern were observed, with more nocturnal records (88.33%; Kruskal-Wallis, p < 0.05). The Chimalapas forest appears to be the second most important terrestrial priority ecoregion, just after the Mayan Forest (Campeche, Chiapas, Quintana Roo), for the conservation of tapir populations, not only for Mexico but also for Central America.


Stage-specific antibody response against two larval stages of Brugia malayi in different clinical spectra of brugian filariasis.
T-cell hypo-responsiveness in microfilaria (Mf) carriers against the microfilarial stage antigen of Brugia malayi has been described, but no study has been carried out to assess antibody dynamics against stage-specific antigens. The work was carried out with the aim to assess stage-specific antibody responses against L3 and microfilarial stage antigens in brugian filariasis in an endemic area. Patients with different clinical spectra of brugian filariasis were recruited to evaluate antibody responses to brugian antigens. Serum samples were collected from patients with different clinical spectra and antibody response was evaluated for total immunoglobulin G (IgG), IgG isotypes (IgG1, IgG2, IgG3, IgG4) and immunoglobulin E (IgE) response to L3 and microfilarial stage by enzyme-linked immunosorbent assay. Paired t-test and one-way analysis of variance were carried out to analyze the data. L3 and microfilarial stage antigens showed almost similar antibody responses in adenolymphangitis (ADL) and chronic pathology (CP) patients, however, diminished antibody response was observed with Mf stage antigen, especially with microfilaraemia. ADL patients had minimum antibody levels of all isotypes except IgG2 on day 0 which showed an increase subsequently, indicating suppression of antibody response during filarial fever. CP patients showed increase in IgE and decrease in IgG4 antibodies on day 365 indicating that these differences may be due to recent conversion into CP. A prominent hyporesponsiveness in microfilaraemic individuals against microfilarial stage, but not against the L3 stage of the same parasite was observed, concluding stage-specificity in humoral immune response in brugian filariasis.


A comparison of grating visual acuity, strabismus, and reoperation outcomes among children with aphakia and pseudophakia after unilateral cataract surgery during the first six months of life.
The method of correcting aphakia after unilateral cataract extraction during infancy is controversial. Some authorities advocate correction with an intraocular lens (IOL) whereas others advocate correction with a contact lens (CL). We compared grating visual acuity, alignment, and reoperative outcomes in age-matched children treated with these 2 modalities at 5 clinical centers. Twenty-five infants born in 1997 or 1998 with a dense unilateral congenital cataract who had cataract surgery coupled with (IOL group, n = 12) or without (CL group, n = 13) primary IOL implantation were enrolled in this study. All patients were prescribed half-time occlusion therapy. In July 1999, their grating visual acuities, ocular alignments, and reoperation rates were assessed. The mean grating visual acuity (LogMAR) for the affected eye was 0.70 +/- 0.32 for the IOL group and 0.87 +/- 0.31 for the CL group (P =.19). The mean interocular difference in grating visual acuity was 0.26 +/- 0.30 for the IOL group and 0.50 +/- 0.28 for the CL group (P =.048). The incidence of strabismus (>10 PD) was 75% in the IOL group compared with 92% in the CL group (P =.24). The incidence of reoperations was 83% in the IOL group compared with 23% in the CL group (P =.003). Our preliminary data suggest that correcting aphakia after unilateral congenital cataract surgery with primary IOL implantation results in an improved visual outcome but a higher rate of complications requiring reoperation. A randomized clinical trial, the Infant Aphakia Treatment Study, is planned to further study the optimal treatment for aphakia following unilateral cataract extraction during infancy.


A comparison of polyamine metabolism in normal and transformed baby-hamster-kidney cells.
Transformed baby-hamster-kidney cells contain higher intracellular concentrations of polyamines than do normal cells. The difference is greater in high-density confluent cultures. Transformed cells incorporate exogenous putrescine into the cells at a faster rate than do normal cells. They also show a marked increase in the rate of spermine biosynthesis compared with normal cells. Transformed cells grown to high cell densities released about 10% of their polyamines into the culture medium in a non-specific manner. In contrast, normal cells, under the same culture conditions, release up to 50% of their intracellular polyamines into the medium almost exclusively as free or conjugated spermidine. The elevated levels of polyamines found in transformed cells therefore appear to be the result of altered transport of polyamines across the cell membrane and of increased rates of biosynthesis.


Peer interaction and loneliness in high-functioning children with autism.
Social interaction with peers and the understanding and feelings of loneliness were examined in 18 high-functioning children with autism and 17 typically developing children matched for IQ, chronological age, gender, and maternal education. Observations were conducted on children's spontaneous social initiations and responses to their peers in natural settings such as recess and snack time, and children reported on their understanding and feelings of loneliness and social interaction. Overall, children with autism revealed a good understanding of both social interaction and loneliness, and they demonstrated a high level of social initiation. However, they spent only half the time in social interactions with peers compared with their matched counterparts, and they interacted more often with a typically developing child than with another special education child. Despite the intergroup differences in frequency of interaction, a similar distribution of interactions emerged for both groups, who presented mostly positive social behaviors, fewer low-level behaviors, and very infrequent negative behaviors. Children with autism reported higher degrees of loneliness than their typical age-mates, as well as a lower association between social interaction and loneliness, suggesting their poorer understanding of the relations between loneliness and social interaction. Research and practice implications of these findings are discussed.


Programmable microfluidic logic device fabricated with a shape memory polymer.
Manipulation of particles in a microfluidic system is an important research subject in biomedical engineering. However, most conventional passive techniques for particle control have difficulties in integrating other functions into microfluidic channels. A unique microfluidic valve was proposed in this study for switchable particle control by employing a shape memory polymer (SMP). A microfluidic logic device can be programmed based on deformation of the SMP microchannel constructed on a poly(dimethylsiloxane) (PDMS) film. Particles in a viscoelastic flow were focused at preferred equilibrium locations by the competition between inertia and elastic forces. The channel shape played an important role in determining those forces in the channel. Hydrodynamic behavior and shape recovery behavior of the SMP microchannel were modeled theoretically. It was confirmed that the particle valve prepared with the SMP implemented a programmable binary logic operation in the microfluidic channel.


The effect of gentamicin on antidiuretic hormone-stimulated osmotic water flow in the toad urinary bladder.
Early gentamicin nephrotoxicity is characterized by a variety of renal transport abnormalities, including polyuria secondary to nephrogenic diabetes insipidus. To investigate whether gentamicin directly interferes with cellular mechanisms responsible for ADH responsivity as a possible contributing factor in this tubular transport alteration, the effect of gentamicin on ADH-stimulated water flow was examined in vitro in the toad urinary bladder. Gentamicin decreased ADH-stimulated osmotic water flow in a dose-dependent manner, with a threshold effect at approximately 0.5 mM. This inhibitory effect occurred only in response to submaximal concentrations of ADH, was reversible, and was preventable with Mg++. Additionally, gentamicin reduced theophylline-stimulated osmotic water flow but had no effect on cyclic AMP-induced water flow. These effects are consistent with a direct gentamicin-related decrease in ADH-responsive adenylate cyclase activity and suggest a mechanism that may contribute to the nephrogenic diabetes insipidus characteristic of early gentamicin nephrotoxicity and the nonoliguric acute renal failure that is a late toxic event occurring with the use of this antibiotic.


Evaluation of knowledge using ExaMe program on the Internet.
In the paper we describe the function of the ExaMe program that serves for evaluation of students' knowledge using Internet. Evaluation is based on the knowledge base of a given course. Two types of evaluation tests are described. The fixed test is appropriate for examination of students by the teacher in computer classroom connected to Internet. By this test the evaluation is done in the limited time period and all students are tested with the same evaluation test. The automatic test is appropriate for self-evaluation and self-study by students on remote places. The student can ask the ExaMe program for the test of different difficulty levels and state the number of questions required. Finally thee first experience with the ExaMe program in practice is given.


Context-Dependent Risk & Benefit Sensitivity Mediate Judgments About Cognitive Enhancement.
Opinions about cognitive enhancement (CE) are context-dependent. Prior research has demonstrated that factors like peer pressure, the influence of authority figures, competition, moral relevance, familiarity with enhancement devices, expertise, and the domain of CE to be enhanced can influence opinions. The variability and malleability of patient, expert, and public attitudes toward CE is important to describe and predict because these attitudes can influence at-home, clinical, research, and regulatory decisions. If individual preferences vary, they could influence opinions about practices and regulations due to disagreements about the desirable levels of risks and benefits. The study of attitudes about CE would benefit from psychological theories that explain judgments. In particular, we suggest that variability in risk and benefit sensitivity could psychologically mediate judgments about CE in many contexts. Drawing from prospect theory, which originated in behavioral economics, it is likely that framing effects, shifted reference points, and the tendency to weigh losses (risks) more heavily than gains (benefits) predict decisions about CE. We suggest that public policy could benefit from a shared conceptual framework, such as prospect theory, that allows us to describe and predict real-world decisions about CE by patients, experts, and the public.


Co-delivery of glycyrrhizin and doxorubicin by alginate nanogel particles attenuates the activation of macrophage and enhances the therapeutic efficacy for hepatocellular carcinoma.
Nanocarrier drug delivery systems (NDDS) have been paid more attention over conventional drug delivery system for cancer therapy. However, the efficacy is hampered by the fast clearance of activated macrophage from the blood circulation system. In this study, glycyrrhizin (GL) was introduced into alginate (ALG) nanogel particles (NGPs) to construct multifunctional delivery vehicle to decrease the fast clearance of activated macrophage and enhance the anticancer efficacy with the combination therapy of GL and doxorubicin (DOX). Methods: We firstly synthesized the GL-ALG NGPs with intermolecular hydrogen bond and ionic bond as the multifunctional delivery vehicle. The immune response and phagocytosis of macrophage on GL-ALG NGPs were investigated on RAW 264.7 macrophages. The pharmacokinetic study of DOX loaded in GL-ALG NGPs was performed in rats. The active targeting effects of GL-ALG NGPs were further studied on hepatocellular carcinoma cell (HepG2) and H22 tumor-bearing mice. Moreover, the anticancer molecular mechanism of DOX/GL-ALG NGPs was investigated on HepG2 cells in vitro and tumor-bearing mice in vivo. Results: GL-ALG NGPs could not only avoid triggering the immuno-inflammatory responses of macrophages but also decreasing the phagocytosis of macrophage. The bioavailability of DOX was increased about 13.2 times by DOX/GL-ALG NGPs than free DOX in blood. The mice with normal immune functions used in constructing the tumor-bearing mice instead of the nude mouse also indicated the good biocompatibility of NGPs. GL-mediated ALG NGPs exhibited excellent hepatocellular carcinoma targeting effect in vitro and in vivo. The results suggested that the anticancer molecular mechanism of the combination therapy of glycyrrhizin and doxorubicin in ALG NGPs was performed via regulating apoptosis pathway of Bax/Bcl-2 ratio and caspase-3 activity, which was also verified in H22 tumor-bearing mice. Conclusion: DOX/GL-ALG NGPs could attenuate the activation of macrophage and enhance the therapeutic efficacy for hepatocellular carcinoma. Our results suggest that the combination therapy would provide a new strategy for liver cancer treatment.


Response of soybean pathogens to glyceollin.
Plants recognize invading pathogens and respond biochemically to prevent invasion or inhibit colonization in plant cells. Enhancing this response in crop plants could improve sustainable methods to manage plant diseases. To enhance disease resistance in soybean, the soybean phytoalexin glyceollin was assessed in soybean hairy roots of two soybean genotypes, Spencer and PI 567374, transformed with either soybean isoflavone synthase (IFS2) or chalcone synthase (CHS6) genes that were inoculated with the soybean pathogens Diaporthe phaseolorum var. meridionales, Macrophomina phaseolina, Sclerotinia sclerotiorum, and Phytophthora sojae. The hairy-root-transformed lines had several-fold decreased levels of isoflavone daidzein, the precursor of glyceollin, and considerably lower concentrations of glyceollin induced by pathogens measured 5 days after fungal inoculation compared with the nontransformed controls without phenolic transgenes. M. phaseolina, P. sojae, and S. sclerotiorum grew much more on IFS2- and CHS6-transformed roots than on control roots, although there was no significant difference in growth of D. phaseolorum var. meridionales on the transformed hairy-root lines. In addition, glyceollin concentration was lower in D. phaseolorum var. meridionales-inoculated transformed and control roots than roots inoculated with the other pathogens. Glyceollin inhibited the growth of D. phaseolorum var. meridionales, M. phaseolina, P. sojae, S. sclerotiorum, and three additional soybean pathogens: Cercospora sojina, Phialophora gregata, and Rhizoctonia solani. The most common product of glyceollin conversion or degradation by the pathogens, with the exception of P. sojae, which had no glyceollin degradation products found in the culture medium, was 7-hydroxyglyceollin.


[Total gastrectomy in the surgery of malignant stomach neoplasms].
An analysis of surgical treatment of 100 patients with malignant tumors of the stomach with using total gastrectomy has shown that postoperative lethality made up 90%, unfavorable results of using total gastrectomy being associated with insufficient sutures of the esophageal-intestinal anastomosis. The modified method used allows to elevate reliability of the esophageal intestinal anastomosis, to shorten the time of operative procedure. Total gastrectomy followed by chemotherapy in the postoperative period prolongs the patient's life, but does not secure against recurrences of the disease in later terms.


The evolution of in vitro fertilization: integration of pharmacology, technology, and clinical care.
For the couple having trouble achieving pregnancy, the options and opportunities for assistance have never been brighter. Options such as controlled ovarian hyperstimulation, in vitro fertilization, and intracytoplasmic sperm injection have been developed over the past five decades and provide hope for couples that previously would have been considered infertile. In vitro fertilization and intracytoplasmic sperm injection represent a coalescence of advances in physiology, endocrinology, pharmacology, technology, and clinical care. In vitro fertilization has assisted well over one million couples in their efforts to start or build a family, and the demand for such services continues to increase. The purpose of this manuscript is to review the pharmacological advances that made controlled ovarian hyperstimulation, and therefore in vitro fertilization and intracytoplasmic sperm injection, possible. We will discuss the early stages of gonadotropin use to stimulate ovarian production of multiple mature eggs, the advances in recombinant technology that allowed purified hormone for therapy, and the use of other hormones to regulate the menstrual cycle such that the likelihood of successful oocyte retrieval and embryo implantation is optimized. Finally, we will review current areas that require particular attention if we are to provide more opportunity for infertile couples.


Three genes preceding pyrE on the Escherichia coli chromosome are essential for survival and normal cell morphology in stationary culture and at high temperature.
Previous studies of the upstream region of the pyrE gene in Escherichia coli revealed three genes of unknown function. Inactivation of these genes (designated orfE, orfX and orfY) by crossing the KmR-cassette-disrupted orf into the chromosome indicated that they were not required during exponential growth (Poulsen et al., Mol., Microbiol., 1989 b). Here we report that the three genes are of importance in the stationary phase. Thus, cultures of the mutants grown to a stationary state in rich media contained bacterial filaments of abnormal morphology. In addition, flow cytometric analyses showed that outgrown cultures of the orf mutants have anomalous size distribution and DNA content, and that rifampicin treatment of exponentially growing mutants results in cell populations with chromosome numbers in the range from about 1 to 10, compared with wild type strains that end up with 4 and 8 full chromosomes. Finally, it appeared that the three orf's are indispensable at high temperatures since the insertion mutants were unable to form colonies above 45 degrees C and since cultures of exponentially growing mutants lysed upon a temperature shift from 37 degrees C to 45 degrees C.


Antimalarials for treating rheumatoid arthritis.
To assess the short-term efficacy and toxicity of antimalarials for the treatment of rheumatoid arthritis (RA). We searched the Cochrane Musculoskeletal Group's trials register, the Cochrane Controlled Trials Register, Medline and Embase up to and including August 2000. We also carried out a handsearch of the reference lists of the trials retrieved from the electronic search. All randomized controlled trials (RCTs) and controlled clinical trials (CCTs) comparing antimalarials against placebo in patients with RA DATA COLLECTION AND ANALYSIS: Data abstraction was carried out independently by two reviewers. The same two reviewers using a validated checklist (Jadad 1996) assessed the methodological quality of the RCTs and CCTs. Rheumatoid arthritis outcome measures were extracted from the publications for the 6-month endpoint. The pooled analysis was performed using standardized mean differences for joint counts, pain and global assessments. Weighted mean differences were used for erythrocyte sedimentation rate (ESR). Toxicity was evaluated with pooled odds ratios for withdrawals. A chi-square test was used to assess heterogeneity among trials. Fixed effects models were used throughout. We found four trials, with 300 patients randomized to hydrochloroquine and 292 to placebo. Only trials evaluating hydroxychloroquine could be pooled in the analysis. A statistically significant benefit was observed when hydroxychloroquine was compared to placebo. The standardized mean differences for the various outcome measures ranged from -0.33 to -0.52, and were statistically significant. Statistically significant differences were also observed for ESR. Overall withdrawals and withdrawals due to lack of efficacy were significantly more frequent in the placebo group. No differences were observed in withdrawals due to toxicity. Hydroxychloroquine appears to be efficacious for the treatment of RA. Its overall effect appears to be moderate, but its low toxicity profile should be considered when treating patients with RA.


Retriever fetches integrins from endosomes.
Recycling from endosomes to the plasma membrane is an important step in cell homeostasis. The retromer/SNX27/WASH complex recycles numerous receptors, but key ones are still unaccounted for. Now a related conserved heterotrimer, called retriever, has been identified that, together with SNX17, the CCC complex and WASH, mediates the recycling of α5β1 integrins.


Comparison of two different blood pumps on delivery of gaseous microemboli during pulsatile and nonpulsatile perfusion in a simulated infant CPB model.
The purpose of this study was to compare two different blood pumps (Jostra roller pump vs. Medos deltastream DP1 rotary pump) on delivery of gaseous microemboli during pulsatile and nonpulsatile perfusion in a simulated infant cardiopulmonary bypass (CPB) model. The Jostra and Medos pump were used in parallel pattern. The circuit was primed with lactated ringer's solution (700 ml) and the postfilter pressure was maintained at 100 mm Hg. Three transducers (postpump, postoxygenator and postfilter sites) of the Emboli Detection and Classification (EDAC) Quantifier were inserted into the CPB circuit to detect and classify gaseous microemboli. Trials were conducted at flow rates ranging from 500 to 1250 ml/min (250 ml/min increments). The arterial filter purge line was kept open during all trials. After injecting 20 ml air into the venous line, 2-minute segments of data were recorded simultaneously through three transducers. This entire process was repeated six times for each unique combination of blood pump, flow rate and perfusion mode, yielding a total of 96 experiments. Independent of perfusion mode and flow rate, Medos pump delivered less gaseous microemboli than Jostra pump at the postpump site, but only at 1,250 ml/min of pump flow rate the differences reached statistical significance (p < 0.01). There was no difference in delivery at the postfilter site. Compared with nonpulsatile flow, pulsatile flow transferred significantly more gaseous microemboli at the postpump site at 1,250 ml/min of pump flow rate in both groups (p < 0.01). The majority of gaseous microemboli were trapped by the membrane oxygenator. The results of this study confirm that rotary pump could deliver less gaseous microemboli than roller pump at the postpump site when a fixed volume air was introduced into the venous line. Pulsatile flow could transfer more gaseous microemboli at the postpump site, no matter which blood pump was used. Only few gaseous microemboli appeared at the postfilter site at high flow rates with an open arterial filter purge line.


The Effects of Social and Behavioral Determinants of Health on the Relationship Between Race and Health Status in U.S. Breast Cancer Survivors.
Background: To examine the association between race and health outcomes among U.S. breast cancer (BC) survivors and explore to what extent do social and behavioral factors contribute to racial disparities for these associations. Materials and Methods: Four hundred forty-one female participants diagnosed with BC in the National Health and Nutrition Examination Survey from 2007 to 2016 were included in this study. Poisson regression with robust variance was used to estimate the prevalence ratio (PrR) and 95% confidence intervals for the associations between race, diabetes, hypertension, and other cancers. Results: The PrR for co-occurrence of diabetes and hypertension was 2.21 (p < 0.001) and 1.62 (p < 0.001) times, respectively, among African Americans (AA) compared with non-Hispanic whites. Body mass index (BMI) explained 17.5% of the association between AA race and diabetes prevalence; a smaller reduction (7.8%) was observed adjusting for type of health insurance coverage, only. A 23.5% reduction was observed in the association between AA race and diabetes prevalence with adjustment for BMI and insurance. The association between AA race and hypertension prevalence was reduced by <6% with addition of individual risk factors, including education, insurance, poverty, obesity, smoking, and physical activity, and with adjustment of the combination of these factors. Conclusions: The association between AA race and diabetes prevalence among BC survivors may be partially explained by BMI and insurance coverage to a lesser extent. Interventions to improve outcomes among AA survivors should focus on weight management strategies.


Maffucci syndrome complicated by three different central nervous system tumors sharing an IDH1 R132C mutation: case report.
Maffucci syndrome (MS) and Ollier disease (OD) are nonhereditary congenital diseases characterized by multiple enchondromas and/or chondrosarcomas. Recent studies have implicated somatic mosaic mutations of isocitrate dehydrogenase 1 or 2 (IDH1/2) as contributing to the pathogenesis of MS and OD. Occasionally, patients with these disorders may also present with central nervous system (CNS) tumors; however, detailed genetic analyses are limited. In this article, the authors report on a male patient with MS, harboring three CNS tumors that share a common genetic alteration. Over a 9-year period, three separate tumor resections were conducted for sellar, intraparenchymal brainstem, and osseous clival tumors. The histopathological diagnoses were pituitary adenoma, diffuse astrocytoma, and chondrosarcoma, respectively. Sanger sequencing revealed a common IDH1 R132C mutation among all three CNS tumors but not in blood DNA. Administering chemotherapy (nimustine) and subsequent radiation therapy to the brainstem glioma and the residual lesion in the clivus have kept the patient progression free for 18 months. This is the first report demonstrating an IDH1 mutation shared among three different CNS tumors in a single patient with MS. The findings support the hypothesis that in MS and OD, a single common IDH1 mutation triggers tumorigenesis in cells of different origins and locations in a somatic mosaic fashion.


Primary care providers' communication with patients during weight counseling: a focus group study.
Primary care providers (PCPs) are encouraged to counsel their obese patients about weight loss. We used focus groups to explore how PCPs communicate with patients about weight management. During the summer of 2010, we conducted five focus groups of community-based PCPs who had patients enrolled in a practice-based, randomized controlled weight loss trial in Maryland. Focus groups were audio-recorded and transcribed verbatim. Two investigators independently coded transcripts for thematic content using editing style analysis. Twenty-six PCPs from six different practices participated. Mean years in practice were 16.4 (SD 11.7) and 77% practiced internal medicine. We identified three communication-based themes about weight loss counseling: (1) motivating patients to lose weight, (2) partnering with the patient to achieve weight loss, and (3) handling challenges that arise during weight counseling. PCPs use a variety of strategies to communicate with their patients about weight loss. Some PCPs already use patient-centered approaches to communicate with their patients about weight loss, suggesting that future weight counseling interventions should be tailored to build upon this strength. PCPs' weight loss counseling may be improved by using techniques with demonstrated behavior change effectiveness such as the 5A's or motivational interviewing.


Lung ultrasound findings in patients with novel SARS-CoV-2.
Over 2 million people worldwide have been infected with severe acute respiratory distress syndrome-coronavirus-2 (SARS CoV-2). Lung ultrasound has been proposed to diagnose and monitor it, despite the fact that little is known about the ultrasound appearance due to the novelty of the illness. The aim of this manuscript is to characterise the lung ultrasonographic appearance of critically ill patients with SARS-CoV-2 pneumonia, with particular emphasis on its relationship with the time course of the illness and clinical parameters. Adult patients from the intensive care unit of two academic hospitals who tested positive for SARS-CoV-2 were included. Images were analysed using internationally recognised techniques which included assessment of the pleura, number of B-lines, pathology in the PLAPS (posterolateral alveolar and/or pleural syndrome) point, bedside lung ultrasound in emergency profiles, and the lung ultrasound score. The primary outcomes were frequencies, percentages and differences in lung ultrasound findings overall and between short (≤14 days) and long (>14 days) durations of symptoms and their correlation with clinical parameters. In this pilot observational study, 61 patients were included with 76 examinations available for analysis. 26% of patients had no anterior lung abnormalities, while the most prevalent pathological ultrasound findings were thickening of the pleura (42%), ≥3 B-lines per view (38%) and presence of PLAPS (74%). Patients with "long" duration of symptoms presented more frequently with a thickened and irregular pleura (32 (21%) versus 11 (9%)), C-profile (18 (47%) versus 8 (25%)) and pleural effusion (14 (19%) versus 3 (5%)), compared to patients with short duration of symptoms. Lung ultrasound findings did not correlate with arterial oxygen tension/inspiratory oxygen fraction ratio, fluid balance or dynamic compliance. SARS-CoV-2 results in significant, but not specific, ultrasound changes, with decreased lung sliding, thickening of the pleura and a B-profile being the most commonly observed. With time, a thickened and irregular pleura, C-profile and pleural effusion become more common findings. When screening patients, a comprehensive ultrasound protocol might be necessary.


Disparities in surgery: access to outcomes.
Surgery is an important intervention to sustain human life through its ability to cure disease, heal fractures, avert maternal death, and provide comfort for those suffering. However, surgical care is unevenly delivered throughout the world. More surprisingly, we have little knowledge about the adequacy and quality of surgical resources globally. The Brigham and Women's Hospital Center for Surgery and Public Health, a joint program of Harvard Medical School and the Harvard School of Public Health, seeks to remedy the knowledge deficit, to provide direction for stakeholders, and direct improvement in public health related to surgical care. To the end of raising awareness of the issue of disparities in surgical care in the United States and the world, an invitational conference on Disparities in Surgical Care: Access to Outcomes was conceived. The Symposium brought together experts from around the world to critically examine this challenging problem. Surgeons, other clinicians, administrators, and health services researchers came together to develop approaches to translate research into practice to address disparities in access and outcomes in surgical care. The synergy of population-based research and clinical practice may allow the surgical healthcare team of the near future to implement strategies to achieve health equity, an important dimension of quality, in surgery.


Mechanisms of epithelial development and neoplasia in the metanephric kidney.
Recent studies on the mechanisms of normal epithelial development in the kidney, and on the aetiology of renal neoplasms, are converging to reveal remarkably close relationships between the phenotypes and behaviours of normally-developing and neoplastic cells. Normal renal epithelia arise from two sources; those of the collecting duct system develop by arborisation of an initially-unbranched ureteric bud, in a manner similar to the development of other glandular organs, while epithelial nephrons develop via an unusual mesenchyme-to-epithelial transition. Both types of development require controlled proliferation, cell-cell and cell-matrix interactions, protease activity etc., but of the two tissues, the development of the nephrons is arguably the more complex. It includes many defined stages, signals and checkpoints that ensure that events happen at the right time, and that processes such as proliferation, apoptosis and differentiation are properly balanced. Detailed investigation of renal neoplasms has revealed some to be caused by mutations in molecules with known roles in normal nephrogenesis (e.g. Wilms' tumour and the WT-1 gene, renal cell carcinoma and the c-met receptor tyrosine kinase gene), some to be caused by mutations in genes expressed during normal development (e.g. renal cell carcinoma and the TSC-2 gene, renal cell carcinoma of the clear cell variety and the VHL gene). Furthermore, these and other tumours of unknown aetiology re-express genes such as Pax-2 that are expressed during the normal mesenchyme-to-epithelium transition but are shut off during terminal differentiation. Their re-appearance in tumours suggests that the cells have 'regressed' in an ontogenic sense, and their biology may therefore be understood most clearly by reference to the properties of normal developing cells rather than cells of a mature kidney.


Childhood mastocytosis.
Important advances have been achieved in recent years in adult mastocytosis. However, our knowledge about childhood mastocytosis is limited because invasive tests are not routinely performed in children. We ignore the frequency of systemic involvement in childhood mastocytosis, its outcome, and which are the main clinical and laboratory parameters associated with persistence into adult mastocytosis and its severity. Childhood mastocytosis is a clonal mast cell disease, with different activating mutations in the KIT gene discovered in most patients. Serum tryptase is the best marker for mast cell burden in children, and, at baseline, correlates well with the severity of symptoms in childhood mastocytosis. Systemic mastocytosis definitely may occur in children, but bone marrow studies to demonstrate a systemic involvement are not routinely performed nor recommended; it can be estimated that around 30% of children may have bone marrow involvement as demonstrated by showing aggregates of mast cells or by flow cytometry of mast cells expressing the aberrant CD25 marker. A new and improved classification of childhood mastocytosis is needed, and should be based on the correlation of clinical manifestations, morphology of mast cells in the skin, and the predicted outcome of the disease. The current classifications of childhood mastocytosis do not address any of these important issues.


Starch extraction from avocado by-product and its use for encapsulation of ginger essential oil by electrospinning.
Starches from alternative sources, such as avocado seed, have potential for application in the encapsulation of essential oils. This study aimed to extract starch from avocado seeds and its use as wall material to encapsulate ginger essential oil (GEO), at different concentrations. The fibers were produced by electrospinning and evaluated by morphology, size, infrared spectra, thermogravimetric properties, contact angle, loading capacity, and antibacterial activity. The major compounds in GEO were α-zingiberene, β-sesquiphellandrene, α-farnesene, and α-curcumene. The starch-GEO fibers presented a higher diameter (∼553 nm) than those without GEO (345 nm). Encapsulation of GEO in starch fibers increased their thermal degradation temperatures from 165.8 °C (free GEO) to 257.6 °C (40 % GEO fibers). The starch-GEO fibers presented characteristic bands of their constituents by infrared spectra. Loading capacity ranged from 44 to 54 %. The fibers showed hydrophilic character, with a contact angle of <90°. Free GEO and the fibers with 50 % of GEO displayed antibacterial activity against Escherichia coli, proving the bioactivity of the starch-GEO fibers and its possible applicability for food packaging. Avocado seed starch showed to be a great wall material for GEO encapsulation.


Metabolic fluxes for nutritional flexibility of Mycobacterium tuberculosis.
The co-catabolism of multiple host-derived carbon substrates is required by Mycobacterium tuberculosis (Mtb) to successfully sustain a tuberculosis infection. However, the metabolic plasticity of this pathogen and the complexity of the metabolic networks present a major obstacle in identifying those nodes most amenable to therapeutic interventions. It is therefore critical that we define the metabolic phenotypes of Mtb in different conditions. We applied metabolic flux analysis using stable isotopes and lipid fingerprinting to investigate the metabolic network of Mtb growing slowly in our steady-state chemostat system. We demonstrate that Mtb efficiently co-metabolises either cholesterol or glycerol, in combination with two-carbon generating substrates without any compartmentalisation of metabolism. We discovered that partitioning of flux between the TCA cycle and the glyoxylate shunt combined with a reversible methyl citrate cycle is the critical metabolic nodes which underlie the nutritional flexibility of Mtb. These findings provide novel insights into the metabolic architecture that affords adaptability of bacteria to divergent carbon substrates and expand our fundamental knowledge about the methyl citrate cycle and the glyoxylate shunt.


The concept of akinesia.
The concept of akinesia deserves to be looked at again in the light of recent work on Parkinsonism, particularly those findings which have resulted from the use of L-dopa in Parkinsonian syndromes. Akinesia and bradykinesia are integral parts of such syndromes, at times even constituting their essential element. Akinesia belongs to a group of psychomotor syndromes, the semiology and pathogenesis of which were the subject of numerous discussions at the beginning of this century. As we have pointed out elsewhere, akinesia cannot be defined solely in terms of its own characteristics: it must be understood equally in its paradoxical aspects--"paradoxical kinesia" in post-encephalitic Parkinsonism in particular, and "paradoxical akinesia" in Parkinsonian patients treated with L-dopa.


Increased Prevalence of Severe Fever with Thrombocytopenia Syndrome in Eastern China Clustered with Multiple Genotypes and Reasserted Virus during 2010-2015.
Severe fever with thrombocytopenia syndrome (SFTS) is a novel tick-borne viral disease with high mortality. Since January 2010, we have conducted an epidemiological surveillance and etiological study of SFTS in Jiangsu and Anhui provinces. From January 2010 through December 2015, a total of 286 SFTS cases were confirmed in Jiangsu and Anhui provinces with a case fatality rate of 16.1%. The majority of confirmed SFTS cases were distributed in the border area of Jiangsu and Anhui provinces. Our findings suggest that the SFTS prevalence rate rose since 2010 and reached its highest in 2015. Phylogenetic analysis demonstrated that the majority of the SFTSV strains (83.6%) from Jiangsu and Anhui provinces belonged to genotypes A and D. Notably, we identified three strains of SFTSV clustered into the genotype E. This is the first report of the genotype E SFTSV strains in mainland of China. A reassortment between genotype A and D was found in the central region of the endemic areas, where three SFTSV genotypes (A, C and D) were co-circulating.


Cell cycle-mediated regulation of hepatic regeneration.
Hepatic regeneration after partial hepatectomy (PH) is characterized by a synchronous induction of normally quiescent hepatocytes to reenter the cell cycle, leading to a complete restoration of hepatic mass. Cell cycle progression requires activation of cyclin-dependent kinases (Cdks) that are regulated by cyclins and Cdk inhibitors. Protein expression of the cyclins (D-type and E), Cdks (Cdk2 and 4), and Cdk inhibitors (p21 and p27) was measured by Western blot after SHAM operation or PH in F344 rats. In addition, Cdk2-associated kinase activity was measured. Rapid induction of D-type and E cyclins, as well as their catalytic partners, Cdk2 and Cdk4, occurred after PH in rats. Complexes containing cyclin E and Cdk2 assembled in the regenerating liver, leading to increased Cdk2-associated kinase activity. The regenerating liver returned to preresection weight by day 7, at which time the Cdk2 activity also returned to SHAM levels. Biphasic induction of the Cdk inhibitor p21 was observed; the first peak occurred as early as 6 hours after PH, with a subsequent peak in expression occurring at 24 to 72 hours after PH. Taken together, these data support the concept that cyclins, Cdks, and Cdk inhibitors regulate cell cycle progression in the regenerating liver. In addition, the induction of p21 at two time points suggests that this protein may regulate both early proliferation and subsequent inhibition of hepatocyte regeneration.


Determination des metabolites monoester de butylbenzyl-phthalate (bbp) par gcms dans les urines de personnes exposees: analysis of the monoester metabolites of butylbenzyl phthalate by gc-ms in urine of exposed workers.
A method of analysis was developed to determine free and glucuronated monobutyl phthalate (BuP) and monobenzyl phthalate (BeP) in urine for the assessment of exposure of man to butylbenzyl phthalate (BBP) in the workplace and in the environment. This method has also been applied in pharmacokinetic studies in experimental animals and the determination in urine of exposed workers. Urine samples are first subjected to enzymatic hydrolysis with ß-glucuronidase to enable the measurement of the total amount of monophthalates excreted. A fraction of the hydrolysate is used for further analysis. Monohexyl phthalate is added as an internal standard and the hydrolysed urine extracted with a n-hexane/ dichloromethane mixture after acidification and saturation with salt. The organic fractions are washed, dehydrated and evaporated. The residue is methylated by means of diazomethane dissolved in diethylether, evaporated and further purified by extraction into n-hexane from an alkaline buffer. The organic fractions are evaporated and the residue redissolved in acetonitrile for analysis by ion trap GC-MS equipped with a 50 m apolar WCOT capillary column. TIC mass chromatograms are recorded from which SIM chromatograms can be derived electronically. The m/z values used are 91, 149, and 163 which provide a sufficient sensitive response and which are specific enough to pick up the methylated monophthalates under investigation. The quantitative limit of detection (LOQ) is 60 μg/L for BuP and BeP when using the Magnum ion Trap detector and 3 μg/L when using the Polaris Q in the splitless mode. The calibration curve in urine is linear from 120 μg/L to 50,000 μg/L with a coefficient of variation of less than 10 % . In case of the Polaris Q linearity started from 10 μg/L. The recovery of the method is monitored by the response signal of the internal standard in the ion chromatogram. In the event of insufficient recovery the analysis is repeated. Variations in recovery are compensated by the internal standard of which the molecular structure is very similar to the ones of the monophthalates under investigation.


An Unexpected Role of Hyaluronic Acid in Trafficking siRNA Across the Cellular Barrier: The First Biomimetic, Anionic, Non-Viral Transfection Method.
Circulating nucleic acids, such as short interfering RNA (siRNA), regulate many biological processes; however, the mechanism by which these molecules enter the cell is poorly understood. The role of extracellular-matrix-derived polymers in binding siRNAs and trafficking them across the plasma membrane is reported. Thermal melting, dynamic light scattering, scanning electron microscopy, and computational analysis indicate that hyaluronic acid can stabilize siRNA via hydrogen bonding and Van der Waals interactions. This stabilization facilitated HA size- and concentration-dependent gene silencing in a CD44-positive human osteosarcoma cell line (MG-63) and in human mesenchymal stromal cells (hMSCs). This native HA-based siRNA transfection represents the first report on an anionic, non-viral delivery method that resulted in approximately 60 % gene knockdown in both cell types tested, which correlated with a reduction in translation levels.


Optimization of Nebulized Budesonide in the Treatment of Acute Exacerbation of Chronic Obstructive Pulmonary Disease.
Clinical studies have suggested nebulized budesonide (NB) as an alternative to systemic corticosteroids for patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). However, the optimal budesonide dose for AECOPD remains unclear. To compare the efficacy and safety of different doses of NB in the management of AECOPD. A total of 321 AECOPD patients with moderate-to-severe exacerbation were randomly divided into three groups and treated with NB. The low dose group (L) was given 4 mg/day (n=95, 1 mg Q6h), while high-dose group 1 (H1, n=111, 2 mg Q6h) and high-dose group 2 (H2, n=115, 4 mg Q12h) were given 8 mg/day. Patients also received routine treatment including oxygen therapy, expectorant, nebulization bronchodilators, antibiotics, and fluid rehydration. The COPD assessment test (CAT), lung function, and artery blood gas were evaluated before and after 3 hrs and 5 days of treatment. In addition, hospital stay, frequency of acute exacerbations within 3 months of discharge, and adverse events during treatment were compared. H1 and H2 showed improved spirograms and CAT score faster than L. In H2, forced expiratory volume in 1 s (FEV1%) at 3 hrs and FEV1%, forced expiratory flow after 50% of the forced vital capacity has been exhaled (FEF50%), mean forced expiratory flow between 25% and 75% of forced vital capacity (FEF25-75%) and CAT score at 5 days were significantly improved compared to L. FEV1% improved most in H2, moderately in H1, and least in L, with significant differences between groups at 5 days. No differences between groups were observed in adverse effects, hospital stay, and frequency of exacerbations within 3 months of discharge. Compared to the conventional dose (4 mg/day), a high dose (8 mg/day) of NB improved pulmonary function and symptoms more effectively in the early treatment of AECOPD, especially when given as 4 mg twice daily.


Promotion of osteogenic cell response using quasicovalent immobilized fibronectin on titanium surfaces: introduction of a novel biomimetic layer system.
Despite the undeniable potential of cell adhesion molecules such as fibronectin to support osteogenic cell responses and consecutive dental implant healing, the most beneficial mode of application onto titanium implant surfaces still requires investigation. Unspecific fibronectin adsorption on titanium dioxide (TiO(2)) surfaces can result in low-loading, high-desorption rates and protein-metal interactions with impaired biologic activity. The aim of the present study was to monitor the osteogenic cell responses (cell adhesion, proliferation, and differentiation) specifically to fibronectin biofunctionalized TiO(2). An innovative biomimetic streptavidin-biotin layer system allows flexible, but stable, specific binding of biotinylated biomolecules such as fibronectin on TiO(2) surfaces. Transparent glass disks were sputtered with TiO(2). The biomimetic layer system was immobilized by self-assembly and consisted of silane, biotin-derivate, streptavidin, and biotinylated fibronectin (bFN). For the control group, unbiotinylated fibronectin was directly coated onto TiO(2). Early cell adhesion dynamics were quantified using automated processing of light microscopy images within the first 24 hours. Relative mRNA expression of integrin-β1, cyclin D1, runt-related gene 2, alkaline phosphatase, and osteocalcin was obtained using quantitative real-time polymerase chain reactions 3 and 7 days after incubation. Although untreated TiO(2) preserved a rather immature osteogenic phenotype, both unbiotinylated fibronectin and bFN promoted osteogenic cell adhesion and cell differentiation. In particular, runt-related gene 2 expression was significantly promoted by bFN after 3 days. In contrast, cyclin D1 expression was decreased for unbiotinylated fibronectin and bFN after 7 days. The introduced biomimetic layer system contributes a coherent immobilization approach of adhesion molecules with promotion of osteogenic cell response in vitro.


Functional Recovery After Rotator Cuff Repair: The Role of Biceps Surgery.
Surgical cuff repair is recommended in a full-thickness rotator cuff tear when nonoperative treatment fails. Surgical cuff repair can include surgery of the long head of the biceps when concomitant biceps pathology is present. However, the studies executed up till now have not yet clearly defined if additional biceps surgery affects the shoulder functionality in patients who underwent rotator cuff repair. To verify if the concomitant biceps surgery prejudices shoulder functionality during the short-term period in rotator cuff repair patients. Prospective and observational study. Outpatient service for rehabilitation. Ninety-three consecutive patients who had undergone surgery for full-thickness symptomatic rotator cuff tear were enrolled for rehabilitation; 25 underwent rotator cuff repair and tendon biceps surgery (ABS), while 68 underwent rotator cuff repair only (RCR). Motor rehabilitation after surgical treatment of rotator cuff repair. Final Constant score was used as primary outcome measure, and efficiency and effectiveness in Constant score were evaluated both at the end of the last cycle of rehabilitation and 6 mo postsurgery. Patients with rotator cuff repair and tendon biceps surgery had lower final scores (36.5 ± 12.0 vs 49.3 ± 13.0, P < .001), effectiveness (40.6 ± 18.0 vs 60.3 ± 20.0, P < .001), and efficiency (0.80 ± 0.5 vs 1.19 ± 0.6, P = .010) in Constant score than those with rotator cuff repair only at the end of rehabilitation. Moreover, they had a lower final score (53.3 ± 14.0 vs 64.5 ± 10.0, P < .001) and effectiveness (66.9 ± 21.0 vs 84.0 ± 16, P < .001) in Constant score 6 mo postsurgery. Gender was a determinant of final score, efficiency, and effectiveness in Constant score at the end of the rehabilitation period, while tendon biceps surgery was a determinant of final score and effectiveness in Constant score at the end of the rehabilitation period and at 6 mo postsurgery. This study highlights that concomitant tendon biceps surgery negatively affects functional outcome of patients who underwent rotator cuff repair and is an important determinant of shoulder functionality in the first 6 mo postsurgery.


Efficiency of N-linked core glycosylation at asparagine-319 of rabies virus glycoprotein is altered by deletions C-terminal to the glycosylation sequon.
In N-linked core glycosylation, the oligosaccharide Glc3Man9GlcNAc2 is transferred to the tripeptide sequon Asn-X-Ser/Thr. However, this process must be regulated by additional protein signals, since many sequons are either poorly glycosylated or not glycosylated at all. Since N-linked glycosylation can influence protein structure and function, understanding these signals is essential for the design and expression of recombinant glycoproteins. Core glycosylation usually occurs cotranslationally in the rough endoplasmic reticulum (RER) during translocation of nascent proteins. Since only regions of a protein immediately near to a sequon or N-terminal to it are thought to be in the RER when core glycosylation occurs, most models predict that regions C-terminal to the sequon do not influence this process. We tested whether regions C-terminal to a sequon can influence its core glycosylation. Full-length (505 amino acid) rabies virus glycoprotein (RGP) mutants, each containing only one of the three sequons normally present in RGP, were used for these studies. Using a cell-free system, the core glycosylation efficiency at each sequon was determined. Termination codons were then introduced into these mutants at defined sites to produce C-terminal truncations, and the effect of each of these truncations on the core glycosylation efficiency at each sequon was assessed. While deletion of the C-terminal transmembrane and cytoplasmic domains did not affect core glycosylation, more extensive C-terminal deletions did result in altered core glycosylation in a site-specific fashion. Specifically, C-terminal truncations resulting in proteins containing 386 or 344 amino acids decreased the efficiency of core glycosylation at Asn319. This demonstrates that core glycosylation efficiency can be influenced by the presence or absence of regions in a protein more than 68 amino acids C-terminal to a specific glycosylation site.


Decreased mRNA expression of several basement membrane components in basal cell carcinoma.
The biologic factors that control the behavior of basal cell carcinoma are poorly understood. This study was undertaken to elucidate the mechanisms responsible for the altered protein levels of several basement membrane components found in basal cell carcinoma. RNA was isolated from papulonodular basal cell carcinoma, normal human epidermal keratinocytes, and normal human skin, reverse transcribed to cDNA and amplified by the polymerase chain reaction utilizing primers specific for the 230 kDa bullous pemphigoid antigen (BPAG1), the 180 kDa bullous pemphigoid antigen (BPAG2), the alpha6 and beta4 chains of the alpha6beta4 integrin complex, and the beta3 chain of laminin 5. Southern blots probed with internal oligonucleotides confirmed that each polymerase chain reaction was specific for the basement membrane component amplified. The mRNA expressions of basement membrane components were indistinguishable between normal human epidermal keratinocytes and normal human skin, and subsequent experiments used normal human epidermal keratinocytes as controls. Quantitation of polymerase chain reaction products indicated that all basement membrane specific mRNA were significantly decreased in basal cell carcinoma as compared with normal human epidermal keratinocytes. The mean polymerase chain reaction product intensities were significantly less in the basal cell carcinoma as compared with the normal human epidermal keratinocytes at the following levels: p < 0.001 for alpha6 and beta4 integrins and the beta3 chain of laminin 5; p < 0.01 for BPAG1; and p < 0.05 for BPAG2. Our results demonstrate that decreased protein levels of basement membrane components in basal cell carcinoma are due at least partially to a downregulation of basement membrane mRNA species. We speculate that these alterations may lead to a structurally incompetent basement membrane that facilitates the basal cell carcinoma ability to invade tissues.