Effects of chromium picolinate on food intake and satiety.
Chromium picolinate (CrPic) has been shown to attenuate weight gain, but the mechanism underlying this effect is unknown. We assessed the effect of CrPic in modulating food intake in healthy, overweight, adult women who reported craving carbohydrates (Study 1) and performed confirmatory studies in Sprague-Dawley rats (Study 2). Study 1 utilized a double-blind placebo-controlled design and randomly assigned 42 overweight adult women with carbohydrate cravings to receive 1,000 mg of CrPic or placebo for 8 weeks. Food intake at breakfast, lunch, and dinner was directly measured at baseline, week 1, and week 8. For Study 2, Sprague-Dawley rats were fasted for 24 h and subsequently injected intraperitoneally with 0, 1, 10, or 50 microg/kg CrPic. Subsequently, rats were implanted with an indwelling third ventricular cannula. Following recovery, 0, 0.4, 4, or 40 ng of CrPic was injected directly into the brain via the intracerebroventricular cannula, and spontaneous 24-h food intake was measured. Study 1 demonstrated that CrPic, as compared to placebo, reduced food intake (P<0.0001), hunger levels (P<0.05), and fat cravings (P<0.0001) and tended to decrease body weight (P=0.08). In study 2, intraperitoneal administration resulted in a subtle decrease in food intake at only the highest dose (P=0.03). However, when administered centrally, CrPic dose-dependently decreased food intake (P<0.05). These data suggest CrPic has a role in food intake regulation, which may be mediated by a direct effect on the brain.


Cost-utility analysis of pralatrexate for relapsed or refractory peripheral T-cell lymphoma based on a case-matched historical control study along with single arm clinical trial.
Patients with relapsed or refractory peripheral T-cell lymphoma (R/R PTCL) treated with pralatrexate have previously shown superior overall survival (OS) compared to those who underwent conventional chemotherapy (CC, 15.4 vs. 4.07 months). We conducted an economic evaluation of pralatrexate from a societal perspective in Korea based on data from the PROPEL phase II study. Using a Markov model with a weekly cycle, we simulated the experience of patients with R/R PTCL receiving pralatrexate or CC for 15 years. The model consists of five health states; initial treatment, treatment pause, subsequent treatment, stem cell transplantation (SCT) success, and death. Comparative effectiveness was based on PROPEL phase II single-arm study and its matched historical control analysis. Costs included drug, drug administration, monitoring, adverse event management, and SCT costs. The incremental cost-effectiveness ratio of the base case was $39,153 per quality-adjusted life-year (QALY) gained. The results of one-way sensitivity analysis ranged from $33,949 to $51,846 per QALY gained, which remained within an implicit willingness-to-pay (WTP) threshold of anticancer drugs in Korea. Pralatrexate is a cost-effective intervention with improved OS and incremental costs within the WTP limit. Pralatrexate could function as a new therapeutic option for patients suffering from life-threatening R/R PTCL.


Ileal recovery of nutrients and mucin in humans fed total enteral formulas supplemented with soy fiber.
The objective of this study was to determine whether soy fiber supplementation of total enteral nutrition formulas affected small intestinal recovery of nitrogen, amino acids, and carbohydrates or mucin output in eight human subjects (four males, four females) with ileostomies. The subjects ingested five test diets to provide 1.0-16.5 g soy fiber/L for 2 consecutive days each. The five test diets, each with a different soy fiber content were formulated by varying the relative proportion (1:0, 0.75:0.25, 0.5:0.5, 0.25:0.75, and 0:1) of two commercially available formulas. Effluent dry matter increased with soy fiber intake as a result of the quantitative recovery of soy fiber nonstarch polysaccharide. Nitrogen and amino acid digestibilities were unchanged by the ingestion of soy fiber. Nutrients from the total enteral nutrition formulas were well digested in the small intestine with true nitrogen and amino acid digestibilities in excess of 90% and starch digestibilities approaching 100%. Ileal mucin output was higher in male subjects and was unaffected by soy fiber intake. In summary, soy fiber supplementation does not compromise protein and carbohydrate absorption from the small intestine of humans.


Myh10 deficiency leads to defective extracellular matrix remodeling and pulmonary disease.
Impaired alveolar formation and maintenance are features of many pulmonary diseases that are associated with significant morbidity and mortality. In a forward genetic screen for modulators of mouse lung development, we identified the non-muscle myosin II heavy chain gene, Myh10. Myh10 mutant pups exhibit cyanosis and respiratory distress, and die shortly after birth from differentiation defects in alveolar epithelium and mesenchyme. From omics analyses and follow up studies, we find decreased Thrombospondin expression accompanied with increased matrix metalloproteinase activity in both mutant lungs and cultured mutant fibroblasts, as well as disrupted extracellular matrix (ECM) remodeling. Loss of Myh10 specifically in mesenchymal cells results in ECM deposition defects and alveolar simplification. Notably, MYH10 expression is downregulated in the lung of emphysema patients. Altogether, our findings reveal critical roles for Myh10 in alveologenesis at least in part via the regulation of ECM remodeling, which may contribute to the pathogenesis of emphysema.


Beyond description: Understanding gender differences in problem gambling.
Background and aims Though women make up roughly one third of all problem gamblers, research has typically focused on male problem gamblers. Recent research has started to shift its attention toward the importance of gender. However, studies rarely attempt to understand gender differences in problem gambling or subject these differences to thorough multivariate analyses. To address some of the gaps in our knowledge of gender differences, we examine whether patterns of gambling behavior and psychological factors mediate the relationship between gender and problem gambling. Methods We use logistic multiple regression to analyze two large Canadian datasets - the 2005 Ontario Prevalence Survey and the 2007 Canadian Community Health Survey. Results Variables found to mediate the relationship between gender and problem gambling are the type(s) of game(s) played (in the 2005 Ontario Prevalence Survey) and the number of games played (in the 2007 Canadian Community Health Survey). Conclusions Men are more likely to be problem gamblers than women, and this gender difference is understandable in terms of differences in patterns of gambling behavior. We conclude that men experience problems because they play riskier games and women experience problems because they prefer chance-based games, which are associated with significantly higher odds of problem gambling. We specify the three main ways that women's reasons for gambling - to escape or for empowerment - translate into chance-based games.


Considerations on isolation rooms and alternative pressure ventilation systems.
The health-care facility environment is involved in disease transmission in essentially two different situations: 1. in cases where patients are immunocompromised and require protection from infections; 2. in cases of inadvertent exposure to environmental or airborne pathogens that can aggravate patients' existent disease and cause illness among health-care personnel. Environmental infection-control strategies and engineering controls can effectively prevent transmission of these infections. In particular the ventilation system is fundamental to the control of the concentration of airborne contaminants within a hospital isolation room because it establishes and maintains appropriate pressure differentials within special care areas of the building. Thus the incidence of health-care-associated infections can be minimized by adherence to ventilation standards suggested in the guidelines for specialized care environments such as Airborne Infection Isolation rooms (AII, as in situation 2 above), and Protective Environments (PE) rooms (as in situation 1 above). This report is a comparative review of the principal guidelines and strategies existing in the international scientific literature for the prevention of environment-associated infections in healthcare facilities using pressure differentials (positive pressure for PE rooms, negative pressure for AII rooms). The purpose of the review is also to investigate the state-of-the-art use of the "alternative pressure rooms", i.e., areas furnished with a ventilation system capable of switching pressure from positive to negative according to patients' needs. The results of the present analysis indicate an unenthusiastic reaction to these "alternative pressure rooms", although there is no scientific evidence against their use.


The assay and properties of labile factor (factor V).
Human oxalated plasma stored at 4 degrees C. until the prothrombin time is increased beyond 60 sec. is a reliable medium for assaying labile factor (factor V) because its response to added labile factor corresponds quantitatively to that of plasma from patients with congenital deficiency of this factor. Such an agreement is not obtained with plasma stored at 37 degrees C. The stability of labile factor is closely associated with ionized calcium. The addition of thrombin to fresh oxalated plasma causes an apparent hyperactivity of labile factor, but this is completely removed by adsorption with Ca(3)(PO)(2). Oxalated plasma when adsorbed with Ca(3)(PO(4))(2) before treatment with thrombin does not develop this adventitious activity, nor does it occur in stored plasma treated with thrombin. The seemingly high labile factor activity in serum can be explained by the activation of this factor which is independent of labile factor but acts synergistically with it. The true labile factor concentration can be determined only after the accelerator is removed by adsorption with Ca(3)(PO(4))(2). A close agreement between the consumption of prothrombin and the loss of labile factor during clotting is observed.


3D-printed composite scaffold with anti-infection and osteogenesis potential against infected bone defects.
In the field of orthopedics, an infected bone defect is a refractory disease accompanied by bone infection and defects as well as aggravated circulation. There are currently no personalized scaffolds that can treat bone infections using local stable and sustained-release antibiotics while providing mechanical support and bone induction to promote bone repair in the process of absorption in vivo. In our previous study, rifampicin/moxifloxacin-poly lactic-co-glycolic acid (PLGA) microspheres were prepared and tested for sustained release and antibacterial activity. The composite scaffold of poly-l-lactic acid (PLLA)/Pearl had a positive effect on mechanics supports and promoted osteogenesis. Therefore, in this study, the personalized scaffolds of PLLA/Pearl were first prepared by 3D printing. Then, rifampicin/moxifloxacin-PLGA (RM-P) microspheres were loaded into the scaffold pores to prepare the PLLA/Pearl/RM-P scaffolds. In this in vitro study, we investigated the structural characteristics and cytocompatibility of 3D-printed composite scaffolds, which indicates the integrity of the components in the scaffolds. The PLLA/Pearl and PLLA/Pearl/RM-P composite scaffolds can promote adhesion, proliferation, and differentiation of human bone marrow mesenchymal stem cells. Moreover, a rabbit model of infected bone defects of the radius was established. PLLA, PLLA/Pearl, and PLLA/Pearl/RM-P scaffolds were implanted into the bone nidus. The therapeutic effect of the three scaffolds on the infected bone defects was evaluated through imaging and microbiological and histological analysis after surgery. Among the three scaffolds, only the PLLA/Pearl/RM-P scaffold had anti-infection and bone defect repair in vivo. 3D printing provides support for personalized scaffold structures, and composite materials ensure that the scaffolds exert anti-infection and bone repair effects. Our study suggests that the PLLA/Pearl/RM-P scaffold is a promising new material in the clinical treatment of infected bone defects.


Epstein-Barr viral genome in lymph nodes from patients with Hodgkin's disease may not be specific to Reed-Sternberg cells.
A possible etiologic role for Epstein-Barr virus (EBV) in Hodgkin's disease (HD) was investigated by probing for EBV genome in 52 biopsy specimens involved with HD and 43 hyperplastic lymph node specimens. Using dot-blot hybridization (Bam HIW probe), Southern blot hybridization (Xho I probe), and polymerase chain reaction analyses, 27%, 27%, and 58% of the nodes with HD were positive for EBV genome, respectively, as compared to 16%, 14%, and 43% in the hyperplastic lymph nodes. Clonal and nonclonal episomal EBV and linear replicating EBV genome were present in both conditions. Immunoglobulin heavy chain gene rearrangements were found in two clonal and two nonclonal EBV-positive HD cases, but not in the lymphoid hyperplasia cases. These findings and other recent reports showing EBV genome in benign lymphoid cells by in situ hybridization in Hodgkin's disease suggest that the characteristics of EBV infection in HD could be explained by the reactive cellular milieu, especially in the setting of defective immunity. The identification of EBV genome in Reed-Sternberg cells may, therefore, be a nonspecific phenomenon.


Implication of the ras and myc oncoproteins in the pathogenesis of mycosis fungoides.
In the present work, we studied the expression of the c-myc oncoprotein p-62 and the ras oncoprotein p-21 in the dermal cellular infiltrate of paraffin embedded skin specimens, obtained from patients suffering from Mycosis Fungoides and Sezary syndrome. Nineteen specimens from early stage Mycosis Fungoides, nineteen from advanced stage Mycosis Fungoides and four from Sezary syndrome were included in the study. The oncoprotein detection was achieved immunohistochemically, using the mouse monoclonal antibody myc 1-9E10 and the rat monoclonal antibody Y13-259 for p-62 and p-21 respectively. Increased detection of both p-62 and p-21 in atypic lymphoid cells was shown in advanced stages of Mycosis Fungoides (third stage plaques and tumors) as compared to early stages (premycotic erythema, second stage plaques). In advanced stages, however, the percentage of P-62+ atypic cells proved to be higher than that of p-21+ atypic lymphoid cells. The implication of increased p-62 and p-21 oncoprotein expression in the process of lymphomagenesis in cutaneous T-cell lymphomas is discussed.


Environmental covariates of Anopheles arabiensis in a rice agroecosystem in Mwea, Central Kenya.
Water quality of aquatic habitats is an important determinant of female mosquito oviposition and successful larval development. This study examined the influence of environmental covariates on Anopheles arabiensis mosquito abundance in the Mwea Irrigation Scheme, Central Province of Kenya, prior to implementation of a malaria vector control program. Experimental rice plots were used to examine the environmental covariates responsible for regulating abundance and diversity of the aquatic stages of malaria vectors. Mosquito larval sampling and water quality analysis were done weekly from the flooding stage to the rice maturation stage. Sampling for mosquito larvae was conducted using standard dipping technique. During each larval collection, environmental covariates such as pH, temperature, conductivity, salinity, dissolved oxygen, water depth, and rice stage were measured. Anopheles arabiensis larval density was highest between 1 wk before transplanting and 4 wk after transplanting with peaks at weeks 0, 3, and 8. The fluctuation in values of the various environmental covariates showed characteristic patterns in different rice growth phases depending on the changes taking place due to the agronomic practices. Using a backward linear regression model, the factors that were found to be associated with abundance of An. arabiensis larvae at any of the rice growing phases included the following: dissolved oxygen, pH, turbidity, water depth, rice height, number of rice tillers, salinity, conductivity, and temperature. The environmental covariates associated with abundance of An. arabiensis were associated with early vegetative stage of the rice growth. For effective control of developmental stages of mosquito larvae, the application of larvicides should be done at the vegetative stage and the larvicides should persist until the beginning of the reproductive stage of the rice.


Online Health Information Seeking Among US Adults: Measuring Progress Toward a Healthy People 2020 Objective.
During the past decade, the availability of health information online has increased dramatically. We assessed progress toward the Healthy People 2020 (HP2020) health communication and health information technology objective of increasing the proportion of health information seekers who easily access health information online. We used data from 4 administrations of the Health Information National Trends Survey (HINTS 2008-2017) (N = 18 103). We conducted multivariable logistic regression analysis to evaluate trends over time in experiences with accessing health information and to examine differences by sociodemographic variables (sex, age, race/ethnicity, education, income, metropolitan status) separately for those who used the internet (vs other information sources) during their most recent search for health information. Among US adults who looked for health information and used the internet for their most recent search, the percentage who reported accessing health information without frustration was stable during the study period (from 37.2% in 2008 to 38.5% in 2017). Accessing information online without frustration was significantly and independently associated with age 35-49 (vs age 18-34) (odds ratio [OR] = 1.34; 95% confidence interval [CI], 1.03 -1.73), non-Hispanic black (vs non-Hispanic white) race/ethnicity (OR = 2.15; 95% CI, 1.55-2.97), and annual household income <$20 000 (vs >$75 000) (OR = 0.66; 95% CI, 0.47-0.93). The percentage of adults who used an information source other than the internet and reported accessing health information online without frustration ranged from 31.3% in 2008 to 42.7% in 2017. Survey year 2017 (vs 2008) (OR = 1.61; 95% CI, 1.09-2.35) and high school graduate education (vs college graduate) (OR = 0.69; 95% CI, 0.49-0.97) were significantly and independently associated with accessing health information without frustration from sources other than the internet. The percentage of online health information seekers reporting easily accessing health information did not meet the HP2020 objective. Continued efforts are needed to enable easy access to online health information among diverse populations.


Characterization of messenger RNA for fructose 1,6-bisphosphate aldolase A isozyme of rat ascites hepatoma AH 7974 cells.
The messenger activity for fructose 1,6-bisphosphate aldolase (EC188.8.131.52) (aldolase) A isozyme has been characterized in the polysome- or the messenger RNA-directed, protein-synthesizing system using the pH 5 fraction of rat liver or wheat germ extracts, respectively. The subunit of aldolase A synthesized in vitro was detected by immunoprecipitation with anti-aldolase A antibody raised in chickens followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The synthesis of the enzyme depended on the addition of polysomes or polyadenylate-containing RNA of rat ascites hepatoma AH 7974 cells which show a complete shift of aldolase isozyme to type A, whereas polysomes of adult rat liver were inactive. The messenger activity for aldolase A was present exclusively on free polysomes but absent on membrane-bound polysomes and in the soluble supernatant fraction of AH 7974 cells. The size of aldolase A messenger RNA determined by formamide-containing sucrose density gradient centrifugation was approximately 5.8 X 10(5) daltons corresponding to 1650 nucleotides. Taking into account the number of amino acid residues in the aldolase A subunit, approximately 400 nucleotides correspond to the noncoding region of aldolase A messenger RNA.


Group A Streptococcal DNase Sda1 Impairs Plasmacytoid Dendritic Cells' Type 1 Interferon Response.
Group A Streptococcus causes severe invasive infections, including necrotizing fasciitis. The expression of an array of virulence factors targeting specific host immune functions impedes successful bacterial clearance. The virulence factor streptococcal DNase Sda1 was previously shown to interfere with the entrapment of bacteria through neutrophil extracellular traps and TLR9 signaling. In this study, we showed that plasmacytoid dendritic cells are recruited to the infected tissue during group A streptococcal necrotizing fasciitis. We found that the streptococcal DNase Sda1 impairs plasmacytoid dendritic cell recruitment by reducing IFN-1 levels at the site of infection. We found that streptococcal DNase Sda1 interferes with stabilization of the DNA by the host molecule HMGB1 protein, which may account for decreased IFN-1 levels at the site of infection.


An organic polymer CuPPc-derived copper oxide as a highly efficient electrocatalyst for water oxidation.
Herein, we report a novel CuPPc (copper polymeric phthalocyanine)/CF (copper foam) nanoflake material, as precatalyst for the generation of an excellent water oxidation catalyst (WOC). Under optimized conditions, the CuPPc-derived Cu oxide affords a current density of 10 mA cm-2 under an overpotential (η) of 287 mV and sustains for at least 50 h in 1.0 M KOH. The strategy presented here is favorable to develop the electrocatalysts for water splitting.


Protein interaction analysis of senataxin and the ALS4 L389S mutant yields insights into senataxin post-translational modification and uncovers mutant-specific binding with a brain cytoplasmic RNA-encoded peptide.
Senataxin is a large 303 kDa protein linked to neuron survival, as recessive mutations cause Ataxia with Oculomotor Apraxia type 2 (AOA2), and dominant mutations cause amyotrophic lateral sclerosis type 4 (ALS4). Senataxin contains an amino-terminal protein-interaction domain and a carboxy-terminal DNA/RNA helicase domain. In this study, we focused upon the common ALS4 mutation, L389S, by performing yeast two-hybrid screens of a human brain expression library with control senataxin or L389S senataxin as bait. Interacting clones identified from the two screens were collated, and redundant hits and false positives subtracted to yield a set of 13 protein interactors. Among these hits, we discovered a highly specific and reproducible interaction of L389S senataxin with a peptide encoded by the antisense sequence of a brain-specific non-coding RNA, known as BCYRN1. We further found that L389S senataxin interacts with other proteins containing regions of conserved homology with the BCYRN1 reverse complement-encoded peptide, suggesting that such aberrant protein interactions may contribute to L389S ALS4 disease pathogenesis. As the yeast two-hybrid screen also demonstrated senataxin self-association, we confirmed senataxin dimerization via its amino-terminal binding domain and determined that the L389S mutation does not abrogate senataxin self-association. Finally, based upon detection of interactions between senataxin and ubiquitin-SUMO pathway modification enzymes, we examined senataxin for the presence of ubiquitin and SUMO monomers, and observed this post-translational modification. Our senataxin protein interaction study reveals a number of features of senataxin biology that shed light on senataxin normal function and likely on senataxin molecular pathology in ALS4.


Prevalence and Molecular Identification of Nematode and Dipteran Parasites in an Australian Alpine Grasshopper (Kosciuscola tristis).
In alpine Australia, Orthoptera are abundant, dominant herbivores, important prey species, and hosts for parasites and parasitoids. Despite the central role of orthopterans in alpine ecosystems, the impact of parasites on orthopteran populations is under-explored. In this study we describe the relationship between parasite prevalence and host sex, body size and year of collection. We accessed an existing, preserved collection of 640 Kosciuscola tristis collected from across its range between 2007 and 2011. Upon dissection we collected juvenile parasites and used molecular tools to identify them to three families (Nematoda; Mermithidae, and Arthropoda: Diptera: Tachinidae and Sarcophagidae). The prevalence of nematodes ranged from 3.5% to 25.0% and dipterans from 2.4% to 20.0%. Contrary to predictions, we found no associations between parasite prevalence and grasshopper sex or size. Although there was an association between prevalence of both nematodes and dipterans with year of collection, this is likely driven by a small sample size in the first year. Our results provide a foundation for future studies into parasite prevalence within the alpine environment and the abiotic factors that might influence these associations.


Engineering in situ cross-linkable and neurocompatible hydrogels.
Physical injuries of the central nervous system (CNS) are prevalent and very severe because the CNS has limited capacity to replace neuronal loss from the injury. A growing body of evidence has suggested that exogenous cell transplantation is one promising strategy to promote CNS regeneration. Direct injection of neural stem cells (NSCs) to the lesion site, however, may not be an optimal therapeutic strategy because of poor viability and functionality of transplanted cells resulting from the local hostile tissue environment. The overall objective of this study is to engineer an injectable and biocompatible hydrogel system as a supportive niche to provide a regeneration permissive microenvironment for transplanted NSCs to survive, functionally differentiate, and integrate with host tissues for CNS regeneration. A highly biocompatible hydrogel, based on thiol functionalized hyaluronic acid and thiol functionalized gelatin (Gtn-SH), which can be cross-linked by poly(ethylene glycol) diacrylate (PEGDA), was used. By controlling the cross-linking density via varying the amount of cross-linker (PEGDA) and the concentration of the adhesive component gelatin, an optimal microenvironment for the survival, proliferation, and neuronal differentiation of NSCs was created in vitro. The soft hydrogel of less than 10 Pa with Gtn-SH content (50%) is one of the optimal conditions to support NSCs growth and neuronal differentiation in vitro. The optimized hydrogel holds great potential as a carrier of stem cells to treat CNS injuries and diseases in which cell therapies may be essential.


[Classifications and scores used in digestive endoscopy].
Interpretation and description of findings detected in upper-endoscopy and colonoscopy are qualitative processes which depend on the experience and skills of the endoscopist performing the procedure. This explains the high variability of endoscopic reports, hampering their interpretation, specially by general practitioners. Classifications, scores and scales give a quantitative support to these qualitative processes. The aim of this review is to describe the classifications, scores and scales most frequently reported in digestive endoscopy, specially those with the highest methodological support in terms of validation and reproducibility. These tools facilitate the description of findings related to gastroesophageal reflux, Barrett's esophagus, gastroesophageal varices, stigmas related to non-variceal gastrointestinal bleeding, advanced and incipient neoplasms, bowel preparation for colonoscopy and severity scores of inflammatory bowel diseases. In summary, these tools enable to standardize endoscopic reports, simplifying their interpretation.


A risk assessment of the effects of mercury on Baltic Sea, Greater North Sea and North Atlantic wildlife, fish and bivalves.
A wide range of species, including marine mammals, seabirds, birds of prey, fish and bivalves, were investigated for potential population health risks resulting from contemporary (post 2000) mercury (Hg) exposure, using novel risk thresholds based on literature and de novo contamination data. The main geographic focus is on the Baltic Sea, while data from the same species in adjacent waters, such as the Greater North Sea and North Atlantic, were included for comparative purposes. For marine mammals, 23% of the groups, each composing individuals of a specific sex and maturity from the same species in a specific study region, showed Hg-concentrations within the High Risk Category (HRC) and Severe Risk Category (SRC). The corresponding percentages for seabirds, fish and bivalves were 2.7%, 25% and 8.0%, respectively, although fish and bivalves were not represented in the SRC. Juveniles from all species showed to be at no or low risk. In comparison to the same species in the adjacent waters, i.e. the Greater North Sea and the North Atlantic, the estimated risk for Baltic populations is not considerably higher. These findings suggest that over the past few decades the Baltic Sea has improved considerably with respect to presenting Hg exposure to its local species, while it does still carry a legacy of elevated Hg levels resulting from high neighbouring industrial and agricultural activity and slow water turnover regime.


Neuroendocrine Dysfunction in a Young Athlete With Concussion: A Case Report.
An 18-year-old female ringette and basketball player presented to our sport concussion clinic 27 months after concussion with fatigue, headache, exercise intolerance, polyuria, nocturia, and difficulties concentrating. Her history was remarkable for 4 previous concussions. Her neurologic examination was normal. Neuroendocrine screen including thyroid function, morning cortisol, glucose, and insulin-like growth factor-1 (screening test for growth hormone deficiency) were normal. Further testing for growth hormone deficiency with an insulin hypoglycemia test revealed severe growth hormone deficiency. Urine and serum electrolytes were borderline normal, suggesting partial diabetes insipidus. Treatments with growth hormone replacement lead to complete recovery. This case highlights the importance of maintaining a high index of suspicion for neuroendocrine abnormalities in athletes with persistent symptoms after sport concussion. Symptoms can be nonspecific and go undiagnosed for years, but appropriate recognition and treatment can restore function.


Epstein-barr virus nuclear antigen 3C activates the latent membrane protein 1 promoter in the presence of Epstein-Barr virus nuclear antigen 2 through sequences encompassing an spi-1/Spi-B binding site.
The Epstein-Barr virus (EBV) nuclear antigen 3C (EBNA-3C) protein is a transcriptional regulator of viral and cellular genes that is essential for EBV-mediated immortalization of B lymphocytes in vitro. EBNA-3C can inhibit transcription through an association with the cellular DNA-binding protein Jkappa, a function shared by EBNA-3A and EBNA-3B. Here, we report a mechanism by which EBNA-3C can activate transcription from the EBV latent membrane protein 1 (LMP-1) promoter in conjunction with EBNA-2. Jkappa DNA-binding sites were not required for this activation, and a mutant EBNA-3C protein unable to bind Jkappa activated transcription as efficiently as wild-type EBNA-3C, indicating that EBNA-3C can regulate transcription through a mechanism that is independent of Jkappa. Furthermore, activation of the LMP-1 promoter is a unique function of EBNA-3C, not shared by EBNA-3A and EBNA-3B. The DNA element through which EBNA-3C activates the LMP-1 promoter includes a Spi-1/Spi-B binding site, previously characterized as an important EBNA-2 response element. Although this element has considerable homology to mouse immunoglobulin light chain promoter sequences to which the mouse homologue of Spi-1 binds with its dimerization partner IRF4, we demonstrate that the IRF4-like binding sites in the LMP-1 promoter do not play a role in EBNA-3C-mediated activation. Both EBNA-2 and EBNA-3C were required for transcription mediated through a 41-bp region of the LMP-1 promoter encompassing the Spi binding site. However, EBNA-3C had no effect on transcription mediated in conjunction with the EBNA-2 activation domain fused to the GAL4 DNA-binding domain, suggesting that it does not function as an adapter between EBNA-2 and the cellular transcriptional machinery. Like EBNA-2, EBNA-3C bound directly to both Spi-1 and Spi-B in vitro. This interaction was mediated by a region of EBNA-3C encompassing a likely basic leucine zipper (bZIP) domain and the ets domain of Spi-1 or Spi-B, reminiscent of interactions between bZIP and ets domains of other transcription factors that result in their targeting to DNA. There are many examples of regulation of the hematopoietic-specific Spi transcription factors through protein-protein interactions, and a similar regulation by EBNA-3C, in conjunction with EBNA-2, is likely to be an important and unique contribution of EBNA-3C to EBV-mediated immortalization.


Irradiation-induced adduct formation of RNA with carcinogenic arylamine derivatives.
Radiolysis of N2O-saturated solutions of transfer RNA (tRNA) and the arylacethydroxamic acids, N-hydroxy-N-2-acetylaminofluorene and N-hydroxy-N-4-acetylaminobiphenyl; their corresponding acetamides, 2-acetylaminofluorene and 4-acetylaminofluorene; or the O-glucuronide of N-hydroxy-N-2-acetylaminofluorene resulted in adduct formation of the nucleic acid with these carcinogenic arylamine derivatives. The yield of adducts on irradiation of the arylacethdroxamic acids with tRNA was greater than that for their corresponding acetamides or the O-glucuronide. The fluorenylacethydroxamic acid and acetamide were also more reactive than the biphenyl analogs. Adduct formation resulting from radiolysis of tRNA and the arylacethydroxamic acids or the O-glucuronide proceeded with retention of both the aromatic nucleus and the N-acetyl group. The yields of adducts were much greater for irradiated mixtures than for irradiation of either component alone followed by mixing. Evaluation of the data shows that initial modification of the tRNA or the carcinogen can lead to adduct formation. In the case of primary radical attack of the nucleic acid, it has been shown that short-lived reactive RNA intermediates are responsible for a major fraction of the observed yield of adducts in the irradiated mixtures. Comparative studies showed that irradiation under conditions that favor reaction of oxidizing radicals enhanced formation of the adducts. Oxygen was shown to protect RNA from irradiation-induced binding of the arylacethydroxamine acids due to competition of O2 with the carcinogen for the reactive RNA intermediates.


The impact of early breaking bad news education: a qualitative study into students' self-reported take-home messages.
This article was migrated. The article was marked as recommended. Introduction: This study investigated the self-reported take-home messages of medical students after an early training module in breaking bad news (BBN). The findings will aid the discussion on how to teach BBN. Methods:We asked 592 second year medical students at two Dutch medical schools to write down what they had learned, right after their first communication skills training in BBN. We analysed these self-reported take-home messages using a hybrid form of qualitative content analysis using SPIKES as a conceptual frame. Results:The most important take-home messages reported by students in both schools were: how to inform the patient (24,5%), how to deal with emotions (20,6%), and how to prepare for a BBN-consultation (16,9%). Additionally, students reported the professional quality "being patient-centered". Conclusions:Early communication skills training on BBN, including true stories and examples of doctors and their BBN-experiences, does not only give the students the knowledge how to break bad news to patients, but also which underlying professional qualities are needed to successfully break bad news. Good role models and realistic examples are therefore important. We propose a new mnemonic PRINSE (PReparation-INformation-Silence-Emotions) for novice learners in the design of a helical curriculum.


The effects of ball possession status on physical and technical indicators during the 2014 FIFA World Cup Finals.
This study examined the effect of high- (HPBPT) and low-percentage ball possession (LPBPT) on physical and technical indicators during 2014 FIFA World Cup matches. This would enable a regression model to be constructed to further understand the impact of different ball possession (BP) strategies on match performance. Data were collected from 346 international soccer players using a multiple-camera computerised tracking system. Although players in HPBPT covered lower distances (P < 0.01) in total and at low speed compared to LPBPT, this produced a trivial effect size (ES). However, they covered similar distances (P > 0.05) at medium and high speeds. Players in LPBPT covered more distance without BP but less with BP than HPBPT (P < 0.01; ES large). All positions in LPBPT spent less time in the opposing half and attacking third than the players in HPBPT (P < 0.01; ES small-moderate), but all positions in HPBPT completed more short and medium passes than LPBPT (P < 0.01; ES moderate). Players in HPBPT produced more solo runs into the attacking third and penalty area than LPBPT (P < 0.05, ES small). The equation to predict BP from physical and technical indicators highlighted the importance of distances covered (total, with and without BP), time spent in the attacking third and successful short passes during matches. In practical terms, high or low BP does not influence the activity patterns of international matches although HPBPT spend more time in offensive areas of the pitch.


Evaluation of the similarities and differences in response patterns to the Pediatric Quality of Life Inventory and the Child Oral Health Impact Scores among youth with cleft.
To examine (a) the pattern of responses to a generic health-related quality of life (HRQL) measure (Pediatric Quality of Life Inventory--PedsQL) and an oral health-related quality of life (OHRQoL) measure (Child Oral Health Impact Profile--COHIP), and (b) the associations of these scores with surgical recommendation status among youth with cleft. Cross-sectional data (baseline) regarding clinicians' surgical recommendations and quality of life (QoL) measures were examined from an ongoing observational study on treatment outcomes. Approximately one-third of the racially and geographically diverse sample (N = 1,200; mean = 11.6 years) received surgical recommendations to correct either visible (aesthetic) or invisible (functional) defects. Effect sizes were used to quantify differences in QoL based on surgical recommendation and to compare the sensitivity of the PedsQL and COHIP subscales. Using Pearson coefficients, the scores of those recommended for surgery were compared with those without a surgical recommendation. A moderate correlation (0.52) was found between the total scores on the PedsQL and COHIP (p < 0.0001). Subscale correlations between the QoL measures ranged from 0.19 to 0.48 with the strongest correlation between the PedsQL Emotional (r = 0.47) and COHIP Socioemotional Well-being subscale. The effect size for the COHIP Socioemotional Well-being (0.39) was larger than the PedsQL Social/Emotional (0.07/0.11) subscale (Z = 5.30/Z = 4.64, p < 0.0001, respectively), and the total COHIP (0.31) was significantly greater than the total PedsQL scale (0.15, z = 2.65, p = 0.008). A significant relationship was found between generic HRQL, OHRQoL, and surgical needs among youth with cleft with the COHIP having larger effect sizes than the PedsQL among surgical groups.


PAM50 Subtypes in Baseline and Residual Tumors Following Neoadjuvant Trastuzumab-Based Chemotherapy in HER2-Positive Breast Cancer: A Consecutive-Series From a Single Institution.
Introduction: HER2-enriched subtype has been associated with higher response to neoadjuvant anti-HER2-based therapy across various clinical trials. However, limited data exist in real-world practice and regarding residual disease. Here, we evaluate the association of HER2-enriched with pathological response (pCR) and gene expression changes in pre- and post-treatment paired samples in HER2-positive breast cancer patients treated outside of a clinical trial. Methods: We evaluated clinical-pathological data from a consecutive series of 150 patients with stage II-IIIC HER2-positive breast cancer treated from August 2004 to December 2012 with trastuzumab-based neoadjuvant chemotherapy. Expression of 105 breast cancer-related genes, including the PAM50 genes, was determined in available pre-and post-treatment formalin-fixed paraffin-embedded tumor samples using the nCounter platform. Intrinsic molecular subtypes were determined using the research-based PAM50 predictor. Association of genomic variables with total pCR was performed. Results: The pCR rate was 53.3%, with higher pCR among hormonal receptor (HR)-negative tumors (70 vs. 39%; P < 0.001). A total of 89 baseline and 28 residual tumors were profiled, including pre- and post-treatment paired samples from 26 patients not achieving a pCR. HER2-enriched was the predominant baseline subtype not only in the overall and HR-negative cohorts (64 and 75%, respectively), but also in the HR-positive cohort (55%). HER2-enriched was associated with higher pCR rates compared to non-HER2-enriched subtypes (65 vs. 31%; OR = 4.07, 95% CI 1.65-10.61, P < 0.002) and this association was independent of HR status. In pre- and post-treatment paired samples from patients not achieving a pCR, a lower proportion of HER2-enriched and twice the number of luminal tumors were observed at baseline, and luminal A was the most frequent subtype in residual tumors. Interestingly, most (81.8%) HER2-enriched tumors changed to non-HER2-enriched, whereas most luminal A samples maintained the same subtype in residual tumors. Conclusions: Outside of a clinical trial, PAM50 HER2-enriched subtype predicts pCR beyond HR status following trastuzumab-based chemotherapy in HER2-positive disease. The clinical value of intrinsic molecular subtype in residual disease warrants further investigation.


Exploiting pleiotropy to map genes for oligogenic phenotypes using extended pedigree data.
We investigated the utility of two approaches for exploiting pleiotropy to search for genes influencing related traits. To do this we first assessed the genetic correlations among a set of five closely related quantitative traits (Q1, Q2, Q3, Q4, Q5). We then used the genetic correlations among these five traits both to remove the common genetic effects of the four remaining traits, thereby identifying the unique genetic contribution to each trait, and to extract a synthetic phenotype which exploits the shared genetic information (pleiotropy) among these five traits. After obtaining these conditional traits, we then searched for evidence of quantitative trait loci (QTLs) (using variance component linkage) influencing the unique residual genetic component for each trait as well as those influencing the expression of the synthetic traits. From this work, we conclude that the removal of the common genetic effects of other traits in a group may be of greater utility when the majority of the pleiotropy initially detected between traits is attributable to the shared additive effects of polygenes, rather than to those of major loci. By contrast, decomposition of the genetic covariance matrix to its principal components is a greater utility when the majority of pleiotropy is attributable to major loci.


Identifying sex-specific risk factors for low bone mineral density in adolescent runners.
Adolescent runners may be at risk for low bone mineral density (BMD) associated with sports participation. Few prior investigations have evaluated bone health in young runners, particularly males. To characterize sex-specific risk factors for low BMD in adolescent runners. Cross-sectional study; Level of evidence, 3. Training characteristics, fracture history, eating behaviors and attitudes, and menstrual history were measured using online questionnaires. A food frequency questionnaire was used to identify dietary patterns and measure calcium intake. Runners (female: n = 94, male: n = 42) completed dual-energy x-ray absorptiometry (DXA) to measure lumbar spine (LS) and total body less head (TBLH) BMD and body composition values, including android-to-gynoid (A:G) fat mass ratio. The BMD was standardized to Z-scores using age, sex, and race/ethnicity reference values. Questionnaire values were combined with DXA values to determine risk factors associated with differences in BMD Z-scores in LS and TBLH and low bone mass (defined as BMD Z-score ≤-1). In multivariable analyses, risk factors for lower LS BMD Z-scores in girls included lower A:G ratio, being shorter, and the combination of (interaction between) current menstrual irregularity and a history of fracture (all P < .01). Later age of menarche, lower A:G ratio, lower lean mass, and drinking less milk were associated with lower TBLH BMD Z-scores (P < .01). In boys, lower body mass index (BMI) Z-scores and the belief that being thinner improves performance were associated with lower LS and TBLH BMD Z-scores (all P < .05); lower A:G ratio was additionally associated with lower TBLH Z-scores (P < .01). Thirteen girls (14%) and 9 boys (21%) had low bone mass. Girls with a BMI ≤17.5 kg/m(2) or both menstrual irregularity and a history of fracture were significantly more likely to have low bone mass. Boys with a BMI ≤17.5 kg/m(2) and belief that thinness improves performance were significantly more likely to have low bone mass. This study identified sex-specific risk factors for impaired bone mass in adolescent runners. These risk factors can be helpful to guide sports medicine professionals in evaluation and management of young runners at risk for impaired bone health.


Oxidative Stress and Male Fertility: Role of Antioxidants and Inositols.
Infertility is defined as a couple's inability to conceive after at least one year of regular unprotected intercourse. This condition has become a global health problem affecting approximately 187 million couples worldwide and about half of the cases are attributable to male factors. Oxidative stress is a common reason for several conditions associated with male infertility. High levels of reactive oxygen species (ROS) impair sperm quality by decreasing motility and increasing the oxidation of DNA, of protein and of lipids. Multi-antioxidant supplementation is considered effective for male fertility parameters due to the synergistic effects of antioxidants. Most of them act by decreasing ROS concentration, thus improving sperm quality. In addition, other natural molecules, myo-inositol (MI) and d-chiro-inositol (DCI), ameliorate sperm quality. In sperm cells, MI is involved in many transduction mechanisms that regulate cytoplasmic calcium levels, capacitation and mitochondrial function. On the other hand, DCI is involved in the downregulation of steroidogenic enzyme aromatase, which produces testosterone. In this review, we analyze the processes involving oxidative stress in male fertility and the mechanisms of action of different molecules.


Homogeneous noncompetitive immunoassay based on the energy transfer between fluorolabeled antibody variable domains (open sandwich fluoroimmunoassay).
The antigen-dependent stabilization of an anti-hen egg lysozyme (HEL) antibody HyHEL-10 variable region was monitored with fluorescence resonance energy transfer (FRET) between fluorolabeled heavy chain (VH) and light chain (VL) fragments. The VH and VL fragments labeled with succinimide esters of fluorescein and rhodamine-X, respectively, were mixed in a cooled cuvette, and the change in fluorescence spectra upon antigen addition was monitored. When excited at 490 nm, significant decrease in the fluorescence at 520 nm and its increase at 605 nm were observed when an increasing amount of HEL was added to the mixture in the concentration range of 1-100 micrograms/mL. The assay, named open sandwich fluoroimmunoassay (FIA), is noncompetitive and homogeneous and can be conducted with one clone of antibody. With the use of appropriate antibodies, it is thought to be a quick and inexpensive alternative to the conventional laborious and/or expensive immunoassays.


Assessment of HER-2 status in invasive breast cancer in Brazil.
To characterize the frequency of HER-2-positive breast cancer in Brazil. In this prospective observational study, we first ascertained the HER-2 status of invasive breast cancer specimens by automated immunohistochemistry (IHC). For specimens classified as 2+ by IHC, we performed in situ hybridization (ISH). From February, 2011 to December, 2012, 1,495 breast specimens were registered, and 1,310 samples collected at 24 centers were analyzed. Median patient age was 54 years, and the majority of samples were obtained from segmental (46.9%) or radical mastectomy (34.4%). The predominant histological type was invasive breast carcinoma of no special type (85%), 64.3% had tubule formation (grade 3), and estrogen/progesterone receptors (ER/PR) were positive in 77.4/67.8% of the specimens analyzed, respectively. Using IHC, we found a negative HER-2 status (0 or 1+) in 72.2% of specimens, and 3+ in 18.5%; the 9.3% scored as 2+ were further analyzed by ISH, of which 15.7% were positive (thus, 20.0% of samples were HER-2- -positive by either method). We found no association between HER-2 scores and menopausal status or histological type. Tumors classified as 3+ came from younger patients, and had higher histological grade and less frequent expression of ER/PR. In the North region of Brazil, 34.7% of samples were 3+, with lower frequencies in the other four regions of the country. Our findings provide estimates for the frequency of HER-2 positivity in Brazil and raise the hypothesis that biological differences may underlie the different distribution of breast-cancer phenotypes among different Brazilian regions.


A single amino acid substitution in the transmembrane envelope glycoprotein of feline immunodeficiency virus alters cellular tropism.
The cellular tropism of the feline immunodeficiency virus (FIV) is affected by changes in variable region 3 (V3) of the surface (SU) envelope glycoprotein (Verschoor, E. J., et al., J. Virol. 69:4752-4757, 1995). By using high-dose DNA transfection, an FIV molecular clone with a non-CRFK-tropic V3 acquired the ability to replicate in CRFK cells. A single point mutation from a methionine to a threonine in the ectodomain of its transmembrane (TM) envelope glycoprotein was responsible for this change in viral tropism. This substitution is located in the putative SU interactive region, between the fusion peptide and the membrane-spanning region. Our results show that this region of the TM envelope glycoprotein constitutes an additional determinant for cell tropism.


Laminitis in young dairy calves fed a high starch diet and with a history of bovine viral diarrhoea virus infection.
Six of the 33 calves born in a Swedish dairy herd during a period of four months developed laminitis when they were eight to 12 weeks old. The clinical signs included difficulty in rising, a stiff gait, overgrown claws and haemorrhages in the sole horn. Samples of blood were taken from four of the calves when they had shown signs of laminitis for two to seven weeks; the serum concentrations of calcium, phosphorus and vitamin D3, the activities of aspartate aminotransferase and glutathione peroxidase, and the patterns of serum proteins were within their normal ranges. The feet of the same four calves were examined after slaughter; the third phalanx of each calf was rotated and its distal end osteolytic. Histologically there was separation and degeneration of the squamous cells of the white line, and thromboses and vasculitis in the fine vessels of the corium. Four of the six affected calves were persistently infected with bovine viral diarrhoea virus and one had antibodies against the virus. From six weeks of age the calves had been fed rye wheat, a hybrid seed rich in starch, and this may have contributed to the outbreak of laminitis.


Cd3P2/Zn3P2 Core-Shell Nanocrystals: Synthesis and Optical Properties.
II-V semiconductor nanocrystals such as Cd3P2 and Zn3P2 have enormous potential as materials in next-generation optoelectronic devices requiring active optical properties across the visible and infrared range. To date, this potential has been unfulfilled due to their inherent instability with respect to air and moisture. Core-shell system Cd3P2/Zn3P2 is synthesized and studied from structural (morphology, crystallinity, shell diameter), chemical (composition of core, shell, and ligand sphere), and optical perspectives (absorbance, emission-steady state and time resolved, quantum yield, and air stability). The improvements achieved by coating with Zn3P2 are likely due to its identical crystal structure to Cd3P2 (tetragonal), highlighting the key role crystallographic concerns play in creating cutting edge core-shell NCs.


Allosterically controlled threading of polymers through macrocyclic dimers.
As part of an ongoing study to construct a molecular Turing machine in which a polymer chain is encoded via allosteric information transfer between macrocyclic complexes, we describe the thermodynamic and kinetic characterization of a multicomponent self-assembled system based on a zinc porphyrin macrocyclic compound, a bidentate ligand (1,4-diazabicyclo[2.2.2]octane, DABCO), and a viologen-substituted polymer guest. Initial addition of DABCO to the porphyrin macrocycle in chloroform solution leads to the formation of a stable 2:1 (porphyrin:DABCO) dimeric complex, even under dilute conditions, by means of strong cooperative interactions involving hydrogen and metal-ligand bonds. Further titration of the porphyrin-DABCO mixtures with the polymer gives rise to a complex array of species in the solution. The system is analyzed in detail by a combination of spectroscopic measurements and computational modeling. Each association constant in the binding scheme and the fraction of each individual complex that is formed in solution are determined precisely using a mass-balance model. Kinetic studies revealed that the rates of the polymer threading and dethreading in and out of the dimeric system are remarkably slow, indicating that the polymer is locked inside the cavity of the stable 2:1 dimeric complex as a result of strong allosteric interactions.


A Graphical Description of Partial Exchangeability.
Partial exchangeability is sufficient for the identification of some causal effects of interest. Here we review the use of common graphical tools and the sufficient component cause model in the context of partial exchangeability. We illustrate the utility of single world intervention graphs (SWIGs) in depicting partial exchangeability and provide an illustrative example of when partial exchangeability might hold in the absence of complete exchangeability.


Competition for resources can promote the divergence of social learning phenotypes.
Social learning occurs when animals acquire knowledge or skills by observing or interacting with others and is the fundamental building block of culture. Within populations, some individuals use social learning more frequently than others, but why social learning phenotypes differ among individuals is poorly understood. We modelled the evolution of social learning frequency in a system where foragers compete for resources, and there are many different foraging options to learn about. Social learning phenotypes diverged when some options offered much better rewards than others and expected rewards changed moderately quickly over time. When options offered similar rewards or when rewards changed slowly, a single social learning phenotype evolved. This held for fixed and simple conditional social learning rules. Sufficiently complex conditional social learning rules prevented the divergence of social learning phenotypes under all conditions. Our results explain how competition can promote the divergence of social learning phenotypes.


Segregated Nanocompartments Containing Therapeutic Enzymes and Imaging Compounds within DNA-Zipped Polymersome Clusters for Advanced Nanotheranostic Platform.
Nanotheranostics is an emerging field that brings together nanoscale-engineered materials with biological systems providing a combination of therapeutic and diagnostic strategies. However, current theranostic nanoplatforms have serious limitations, mainly due to a mismatch between the physical properties of the selected nanomaterials and their functionalization ease, loading ability, or overall compatibility with bioactive molecules. Herein, a nanotheranostic system is proposed based on nanocompartment clusters composed of two different polymersomes linked together by DNA. Careful design and procedure optimization result in clusters segregating the therapeutic enzyme human Dopa decarboxylase (DDC) and fluorescent probes for the detection unit in distinct but colocalized nanocompartments. The diagnostic compartment provides a twofold function: trackability via dye loading as the imaging component and the ability to attach the cluster construct to the surface of cells. The therapeutic compartment, loaded with active DDC, triggers the cellular expression of a secreted reporter enzyme via production of dopamine and activation of dopaminergic receptors implicated in atherosclerosis. This two-compartment nanotheranostic platform is expected to provide the basis of a new treatment strategy for atherosclerosis, to expand versatility and diversify the types of utilizable active molecules, and thus by extension expand the breadth of attainable applications.


Systematic review on the use and description of measures to evaluate psychotherapeutic interventions with children and adolescents in Brazil.
Psychotherapeutic interventions in childhood and adolescence are recognized as a mental treatment and as a tool to reduce psychological disorders in youth and adulthood. Therefore, there is an increasing concern about evidence of effectiveness of mental disorder interventions and adequacy of measurement. The objectives of this systematic review were to investigate predominant research designs and instruments used in Brazilian studies describing psychotherapeutic interventions with children and adolescents and to examine how these instruments are described regarding evidence of validity and reliability. Five databases were surveyed, and for each one two independent judges performed the selection of records and applied the inclusion and exclusion criteria. The final database comprised 28 papers, in which 92 instruments were referred. Fifty-seven instruments cited did not have descriptions of evidence of validity for the Brazilian population; for 31 instruments, validity evidence was mentioned, but the study did not detail which validity parameter was used; three studies described content validity evidence for their instruments. Furthermore, information about reliability was described for only two instruments. A lack of studies in the field of child and adolescent psychotherapy was found in Brazil. There is a significant need for the field to attend both the psychometric properties and the quality of description of research instruments. The scientific production of studies focused on the evaluation of psychotherapeutic interventions may promote evidence-based psychotherapy and justify the offer of mental treatment in different contexts.


Effects of temperature and fluid media on the scroll width size of the Descemet's membrane endothelial keratoplasty (DMEK) donor graft.
Our study was conducted to evaluate whether higher temperature leads to increased - or wider - scroll widths of the Descemet's membrane endothelial keratoplasty (DMEK) donor graft. To investigate the effects of temperature and fluid media on the DMEK donor graft scroll widths. This research work was a laboratory investigation. Baseline cell count was taken via specular microscopy for the donor corneas at room temperature (20°C-25°C). The endothelium sides of the donor corneas were stained with Trypan Blue Solution 0.4% for 30 s, and the Descemet's membranes were stripped. The DMEK donor grafts were placed into three different fluid media - Optisol®, Balanced Salt Solution (BSS), and BSS PLUS® (BSS Plus). The DMEK donor grafts were then transferred into cold temperature (4°C) for 60 min, after which the donor grafts' scroll widths were examined and measured. The donor grafts were then warmed in the incubator and brought to physiological temperature (35°C-37°C), and their scroll widths were examined and measured again. In 30 measurements of ten tissues across three temperature and fluid conditions, the average scroll width measured 1.73 mm, ranging from 1.1 to 2.9 mm. In a mixed linear model, the scroll widths increased with temperature (P=0.02). There was no significant difference in scroll widths among the three solutions (P=0.84, mixed linear model). We observed an increase in DMEK donor graft scroll widths with higher temperatures. The usage of BSS Plus as media solution could also lead to smaller DMEK donor graft scroll widths, compared with BSS, but our study does not establish this.


The thoracic surgeon's role in the management of patients with HIV infection and AIDS.
The advent of improved medical therapy with multidrug HIV medicines including protease inhibitors has prolonged life expectation of patients with HIV infection. The risk of perinatal transmission has also decreased with education and antiviral medications. Education of all groups about precautions to decrease HIV transmission appears to have resulted in a decline in the disease incidence; however, this trend has not been mirrored in other countries. All of these factors combine to increase the probability that the surgeon's role to take care of patients with HIV-related illnesses could be significant in the future. The thoracic surgeon may be called upon to assist in the diagnosis and treatment of HIV-infected patients with thoracic complications including pulmonary, cardiac, and esophageal problems. The morbidity and mortality of procedures performed on these patients appear to be no different than patients without HIV. We use very aggressive treatment strategies in patients with AIDS and HIV infections. The use of VATS, especially in the treatment of pneumothoraces and empyemas, should be used and has been shown to be safe and efficacious.


Demographic and clinical features of child abuse and neglect cases.
The aim of this study was to present child abuse and neglect (CAN) cases of the Child and Adolescent Psychiatry Department of Hacettepe University Faculty of Medicine. The charts of all the patients seen at the Child and Adolescent Psychiatry Clinic between 2000 and 2004 were screened regarding the presence of CAN. The mean age of the children was 10.9 years. The offender was generally an extra-familial acquaintance (40.7%). The most common type of abuse was sexual (77.8%) and the most common associated types of abuse were physical and sexual (7.4%). Attention deficit hyperactivity disorder (ADHD) was observed as the most common psychiatric diagnosis (22.2%). Abuse types in relation to age and gender of abused child, risk factors and associated psychopathologies are discussed. Issues related to legislative process and ecology of the abuse experience are mentioned as restrictions for the prevention and treatment of CAN.


The immune response of the teleost, Tilapia mossambica to soluble and cellular antigens.
The immune response of Tilapia mossambica to bovine serum albumin (BSA) and sheep red blood cells (SRBC) was characterized in detail in terms of the appearance of hemolysin plaque-forming cells and circulating antibodies at 30 degrees C. Plaque-forming cells (PFC) were detected in the spleen, head-kidney and thymus of immunized fish and the maximum number was observed in these organs on the fifth day after immunization with SRBC. Peak circulating antibody response occurred on day 8 for SRBC and on day 11 for BSA. Following the second injection of the same antigen, a specific anamnestic response was observed with increased production of PFC and serum antibody. No cross reactivity was found when anti-SRBC antibody was tested with rat erythrocytes. Tests with 2-mercaptoethanol showed that all of the agglutinating antibody produced after both the first and second injection was mercaptoethanol sensitive. Analysis of histological and smear preparations revealed that there were consistent cellular changes occurring in the spleen as well as the head-kidney due to immunization.


Lack of antiviral activity of probenecid in vitro and in Syrian golden hamsters.
Antiviral interventions are required to complement vaccination programmes and reduce the global burden of COVID-19. Prior to initiation of large-scale clinical trials, robust preclinical data to support candidate plausibility are required. This work sought to further investigate the putative antiviral activity of probenecid against SARS-CoV-2. Vero E6 cells were preincubated with probenecid, or control media for 2 h before infection (SARS-CoV-2/Human/Liverpool/REMRQ0001/2020). Probenecid or control media was reapplied, plates reincubated and cytopathic activity quantified by spectrophotometry after 48 h. In vitro human airway epithelial cell (HAEC) assays were performed for probenecid against SARS-CoV-2-VoC-B.1.1.7 (hCoV-19/Belgium/rega-12211513/2020; EPI_ISL_791333, 2020-12-21) using an optimized cell model for antiviral testing. Syrian golden hamsters were intranasally inoculated (SARS-CoV-2 Delta B.1.617.2) 24 h prior to treatment with probenecid or vehicle for four twice-daily doses. No observable antiviral activity for probenecid was evident in Vero E6 or HAEC assays. No reduction in total or subgenomic RNA was observed in terminal lung samples (P > 0.05) from hamsters. Body weight of uninfected hamsters remained stable whereas both probenecid- and vehicle-treated infected hamsters lost body weight (P > 0.5). These data do not support probenecid as a SARS-CoV-2 antiviral drug.


Extrapolating Parametric Survival Models in Health Technology Assessment: A Simulation Study.
Extrapolations of parametric survival models fitted to censored data are routinely used in the assessment of health technologies to estimate mean survival, particularly in diseases that potentially reduce the life expectancy of patients. Akaike's information criterion (AIC) and Bayesian information criterion (BIC) are commonly used in health technology assessment alongside an assessment of plausibility to determine which statistical model best fits the data and should be used for prediction of long-term treatment effects. We compare fit and estimates of restricted mean survival time (RMST) from 8 parametric models and contrast models preferred in terms of AIC, BIC, and log-likelihood, without considering model plausibility. We assess the methods' suitability for selecting a parametric model through simulation of data replicating the follow-up of intervention arms for various time-to-event outcomes from 4 clinical trials. Follow-up was replicated through the consideration of recruitment duration and minimum and maximum follow-up times. Ten thousand simulations of each scenario were performed. We demonstrate that the different methods can result in disagreement over the best model and that it is inappropriate to base model selection solely on goodness-of-fit statistics without consideration of hazard behavior and plausibility of extrapolations. We show that typical trial follow-up can be unsuitable for extrapolation, resulting in unreliable estimation of multiple parameter models, and infer that selecting survival models based only on goodness-of-fit statistics is unsuitable due to the high level of uncertainty in a cost-effectiveness analysis. This article demonstrates the potential problems of overreliance on goodness-of-fit statistics when selecting a model for extrapolation. When follow-up is more mature, BIC appears superior to the other selection methods, selecting models with the most accurate and least biased estimates of RMST.


Blood pressure predicts risk of developing end-stage renal disease in men and women.
Blood pressure as a risk factor for development of end-stage renal disease has not been fully studied, particularly in women. We studied the development of end-stage renal disease from 1983 through 2000 in 98 759 subjects, 46 881 men and 51 878 women, 20 to 98 years of age, who were screened in 1983 in Okinawa, Japan. Data for all dialysis patients registered from 1983 to 2000 in Okinawa were used to identify the screened subjects in whom end-stage renal disease developed. In follow-up, 400 subjects, 231 men and 169 women, had end-stage renal disease. Age, body mass index, and adjusted relative risk for systolic and diastolic blood pressure for both men and women were measured. When these results were compared with an optimal blood pressure, the relative risk of development of end-stage renal disease for those with high-normal blood pressure and hypertension were significant in both men and women. Hypertension is a significant risk factor for development of end-stage renal disease not only in men but also in women. Control of blood pressure within normal levels should be stressed as a strategy to prevent end-stage renal disease in both men and women.


Kinetic analysis of human PrimPol DNA polymerase activity reveals a generally error-prone enzyme capable of accurately bypassing 7,8-dihydro-8-oxo-2'-deoxyguanosine.
Recent studies have identified human PrimPol as a new RNA/DNA primase and translesion DNA synthesis polymerase (TLS pol) that contributes to nuclear and mitochondrial DNA replication. We investigated the mechanism of PrimPol polymerase activity on both undamaged and damaged DNA substrates. With Mg²⁺ as a cofactor, PrimPol binds primer-template DNA with low affinity K(d,DNA) values (∼200-1200 nM). DNA binding is enhanced 34-fold by Mn²⁺ (K(d,DNA) = 27 nM). The pol activity of PrimPol is increased 400-1000-fold by Mn²⁺ compared to Mg²⁺ based on steady-state kinetic parameters. PrimPol makes a mistake copying undamaged DNA once every ∼100-2500 insertions events, which is comparable to other TLS pols, and the fidelity of PrimPol is ∼1.7-fold more accurate when Mg²⁺ is the cofactor compared to Mn²⁺. PrimPol inserts dCMP opposite 8-oxo-dG with 2- (Mn²⁺) to 6-fold (Mg²⁺) greater efficiency than dAMP misinsertion. PrimPol-catalyzed dCMP insertion opposite 8-oxo-dG proceeds at ∼25% efficiency relative to unmodified template dG, and PrimPol readily extends from dC:8-oxo-dG base pairs (bps) with ∼2-fold greater efficiency than dA:8-oxo-dG bps. A tetrahydrofuran (THF) abasic-site mimic decreases PrimPol activity to ∼0.04%. In summary, PrimPol exhibits the fidelity typical of other TLS pols, is rather unusual in the degree of activation afforded by Mn²⁺, and accurately bypasses 8-oxo-dG, a DNA lesion of special relevance to mitochondrial DNA replication and transcription.


[Intestinal dysbacteriosis in different age groups of residents of Kemerovo].
The state of microflora of the large intestine in 877 persons of different age groups in Kemerovo was studied. The study revealed that intestinal dysbacteriolysis of various degrees of severity was rather widely spread among the residents of the city. The main groups of risk, found to comprise young children, adolescents and people above 60 years of age, were established.


Effect of vitamin D deficiency on the 75 g oral glucose tolerance test screening and insulin resistance.
Gestational diabetes mellitus (GDM), is the most common medical complications of pregnancy. This study aimed to clarify the effect of second-trimester vitamin D deficiency on the 75 g oral glucose tolerance test (OGTT) screening and insulin resistance. A total of 120 pregnant women with a singleton pregnancy at a gestational age of 26-28 weeks were analyzed. Participants were divided into two groups according to 25-hydroxyvitamin D levels; vitamin D deficiency, and control groups. For GDM scan, 75 g OGTT was preferred. GDM prevalence was 17.5% in vitamin D deficiency group and 13.75% in control group, there is no significant difference in GDM prevalence (p = 0.149). Fasting plasma glucose and 1-h plasma glucose levels were significantly higher in the vitamin D deficiency group than in the control group (p < .001 and p < .001, respectively). No significant differences were observed between 2-hour plasma glucose levels (p = .266). The HOMA-IR level was significantly higher in the vitamin D deficiency group than in the control group (p < .001). The findings of the present study suggested that vitamin D deficiency in the second trimester was inversely correlated with fasting and 1-h plasma glucose after 75 g glucose challenge test; also, low 25 OHD3 levels were associated with insulin resistance.


Structural basis for subversion of host cell actin cytoskeleton during Salmonella infection.
Secreted bacterial type III secretion system (T3SS) proteins are essential for successful infection by many human pathogens. Both T3SS translocator SipC and effector SipA are critical for Salmonella infection by subversion of the host cell cytoskeleton, but the precise molecular interplay between them remains unknown. Here, using cryo-electron microscopy, we show that SipA binds along the F-actin grooves with a unique binding pattern. SipA stabilizes F-actin through charged interface residues and appears to prevent inorganic phosphate release through closure of the "back door" of adenosine 5'-triphosphate pocket. We also show that SipC enhances the binding of SipA to F-actin, thus demonstrating that a sequential presence of T3SS proteins in host cells is associated with a sequence of infection events-starting with actin nucleation, filament growth, and stabilization. Together, our data explain the coordinated interplay of a precisely tuned and highly effective mechanism during Salmonella infection and provide a blueprint for interfering with Salmonella effectors acting on actin.


The Influence of Diet on Fertility and the Implications for Public Health Nutrition in the United States.
Despite growing evidence of the impact of diet on human fertility, few studies have examined the public health implications of this association in the United States (U.S.). This narrative review summarizes current scientific evidence on associations between dietary intake and fertility, discusses challenges in the public health landscape surrounding infertility, and proposes evidence-based recommendations to address these issues. Diets high in unsaturated fats, whole grains, vegetables, and fish have been associated with improved fertility in both women and men. While current evidence on the role of dairy, alcohol, and caffeine is inconsistent, saturated fats, and sugar have been associated with poorer fertility outcomes in women and men. Furthermore, women and men with obesity [body mass index (BMI) ≥ 30 kg/m2] have a higher risk of infertility. This risk is extended to women who are underweight (BMI <20 kg/m2). Diet and BMI influence outcomes during clinical treatment for infertility. Further, women in the U.S. who belong to an underrepresented minority group, have low income, or have low educational attainment, have significantly higher rates of infertility outcomes as compared to women who are non-Hispanic white, have high income, or have high educational attainment. Given this, it may be prudent to integrate nutrition counseling into both clinical guidelines for infertility as well as national dietary guidelines for individuals of reproductive age. Further studies on diet and reproductive health may enhance our ability to improve existing fertility programs across the U.S. and to deliver tailored care to women and men within at-risk groups.


High levels of adenosine deaminase on dendritic cells promote autoreactive T cell activation and diabetes in nonobese diabetic mice.
Adenosine has been established as an important regulator of immune activation. It signals through P1 adenosine receptors to suppress activation of T cells and professional APCs. Adenosine deaminase (ADA) counters this effect by catabolizing adenosine. This regulatory mechanism has not been tested in a disease model in vivo. Questions also remain as to which cell types are most sensitive to this regulation and whether its dysregulation contributes to any autoimmune conditions. We approached this issue using the NOD model. We report that ADA is upregulated in NOD dendritic cells, which results in their exuberant and spontaneous activation. This, in turn, triggers autoimmune T cell activation. NOD DCs deficient in ADA expression have a greatly reduced capacity to trigger type I diabetes. We also provide evidence that although many cell types, particularly T cells, have been implicated as the suppression targets by adenosine in an in vitro setting, DCs also seem to be affected by this regulatory mechanism. Therefore, this report illustrates a role of ADA in autoimmunity and suggests a potential target for therapeutic intervention.


[Reliability and validity of Health Scale of Traditional Chinese Medicine].
To test the reliability and validity of Health Scale of Traditional Chinese Medicine (HSTCM) by means of questionnaires. A cross-sectional survey was conducted at Liwan Community of Guangzhou, Old People's Home in Guangzhou and Outpatient Department of the Second Affiliated Hospital of Guangzhou University of Chinese Medicine. A total of 652 Chinese individuals (over 18 years old) were assessed with the 88-item version of HSTCM and World Health Organization Quality of Life Measure-Abbreviated Version (WHOQOL-BREF), which were randomly delivered to them. Some socioeconomic characteristics were registered. A test-retest reliability (15-day interval) was found among the 76 persons who completed the questionnaires by themselves. Intra-class correlation coefficient (ICC) was 0.93. Associated 95% confidence interval (CI) was 0.89-0.96. Split-half reliability was 0.79. Inter-investigator reliability (0.93) was also good, and the ICC of HSTCM was 0.90 (95%CI 0.67-0.97). The correlation between HSTCM and WHOQOL-BREF was -0.66. The correlations of HSTCM and questionnaire deliver order, investigator, interview date and interview time were 0.06, -0.12, -0.17 and 0.20 respectively. The correlation between HSTCM and self-rated health (0.46) was greater than that between HSTCM and chronic illness (0.28). Divided by individuals with or without chronic illness, area under the ROC (receiver operator characteristic) curve for HSTCM was 0.67 (95%CI 0.63-0.71). It indicates that the HSTCM is conceptually valid with satisfactory psychometric properties and forms a basis for further applications in clinical research of traditional or integrative medicine.


PRC2 engages a bivalent H3K27M-H3K27me3 dinucleosome inhibitor.
A lysine-to-methionine mutation at lysine 27 of histone 3 (H3K27M) has been shown to promote oncogenesis in a subset of pediatric gliomas. While there is evidence that this "oncohistone" mutation acts by inhibiting the histone methyltransferase PRC2, the details of this proposed mechanism nevertheless continue to be debated. Recent evidence suggests that PRC2 must simultaneously bind both H3K27M and H3K27me3 to experience competitive inhibition of its methyltransferase activity. In this work, we used PRC2 inhibitor treatments in a transgenic H3K27M cell line to validate this dependence in a cellular context. We further used designer chromatin inhibitors to probe the geometric constraints of PRC2 engagement of H3K27M and H3K27me3 in a biochemical setting. We found that PRC2 binds to a bivalent inhibitor unit consisting of an H3K27M and an H3K27me3 nucleosome and exhibits a distance dependence in its affinity for such an inhibitor, which favors closer proximity of the 2 nucleosomes within a chromatin array. Together, our data precisely delineate fundamental aspects of the H3K27M inhibitor and support a model wherein PRC2 becomes trapped at H3K27M-H3K27me3 boundaries.


Chiari III malformation in a neonate.
Chiari malformation (CM) is a group of complex deformities of the posterior fossa and hindbrain, of which CMIII is the rarest. We report a term neonate, with an antenatal diagnosis of occipital encephalocele, who underwent resection of the encephalocele and ligation of vessels, with repair of a large scalp defect and dural reconstruction on day 4 of life. The parents of the child had been counselled for a guarded and poor prognosis on initial diagnosis. The child has had a good postoperative course without complications but suffers from cortical visual impairment and global developmental delay.


The Influence of Dual Pressure Biofeedback Units on Pelvic Rotation and Abdominal Muscle Activity during the Active Straight Leg Raise in Women with Chronic Lower Back Pain.
[Purpose] This study was performed to assess the influence of applying dual pressure biofeedback units (DPBUs) on the angle of pelvic rotation and abdominal muscle activity during the active straight leg raise (ASLR). [Subjects] Seventeen patients with low-back pain (LBP) participated in this study. [Methods] The subjects were asked to perform an active straight leg raise (ASLR) without a PBU, with a single PBU, and with DPBUs. The angles of pelvic rotation were measured using a three-dimensional motion-analysis system, and the muscle activity of the bilateral internal oblique abdominis (IO), external oblique abdominis (EO), and rectus abdominis (RA) was recorded using surface electromyography (EMG). One-way repeated-measures ANOVA was performed to determine the rotation angles and muscle activity under the three conditions. [Results] The EMG activity of the ipsilateral IO, contralateral EO, and bilateral RA was greater and pelvic rotation was lower with the DPBUs than with no PBU or a single PBU. [Conclusion] The results of this study suggest that applying DPBUs during ASLR is effective in decreasing unwanted pelvic rotation and increasing abdominal muscle activity in women with chronic low back pain.


Non-invasive Ventilation for Children With Chronic Lung Disease.
Advances in medical care and supportive care options have contributed to the survival of children with complex disorders, including children with chronic lung disease. By delivering a positive pressure or a volume during the patient's inspiration, NIV is able to reverse nocturnal alveolar hypoventilation in patients who experience hypoventilation during sleep, such as patients with chronic lung disease. Bronchopulmonary dysplasia (BPD) is a common complication of prematurity, and despite significant advances in neonatal care over recent decades its incidence has not diminished. Most affected infants have mild disease and require a short period of oxygen supplementation or respiratory support. However, severely affected infants can become dependent on positive pressure support for a prolonged period. In case of established severe BPD, respiratory support with non-invasive or invasive positive pressure ventilation is required. Patients with cystic fibrosis (CF) and advanced lung disease develop hypoxaemia and hypercapnia during sleep and hypoventilation during sleep usually predates daytime hypercapnia. Hypoxaemia and hypercapnia indicates poor prognosis and prompts referral for lung transplantation. The prevention of respiratory failure during sleep in CF may prolong survival. Long-term oxygen therapy has not been shown to improve survival in people with CF. A Cochrane review on the use NIV in CF concluded that NIV in combination with oxygen therapy improves gas exchange during sleep to a greater extent than oxygen therapy alone in people with moderate to severe CF lung disease. Uncontrolled, non-randomized studies suggest survival benefit with NIV in addition to being an effective bridge to transplantation. Complications of NIV relate mainly to prolonged use of a face or nasal mask which can lead to skin trauma, and neurodevelopmental delay by acting as a physical barrier to social interaction. Another associated risk is pulmonary aspiration caused by vomiting whilst wearing a face mask. Adherence to NIV is one of the major barriers to treatment in children. This article will review the current evidence for indications, adverse effects and long term follow up including adherence to NIV in children with chronic lung disease.


[Volumetry of abdominal tumors. Problems--feasibility].
If multimodal tumor therapy in the abdomen is to be effective, exact staging is required for which one needs precise planimetric and volumetric data. The most important indication for clinically reliable volumetric determination of tumor size in the abdominal region is monitoring liver metastases during chemotherapy. Determination of volume can be effectively realized using 3D reconstruction. Therefore, the primary data set must be complete and contiguous. The mass should be depicted strongly enhanced and free of artifacts. At present, this prerequisite can only be complied with using thin-slice spiral CT. Phantom studies have proven that a semiautomatic reconstruction algorithm is recommendable. The basic difficulties involved in volumetric determination of tumor size are the problems in differentiating active malignant mass and changes in the surrounding tissue, as well as the lack of histomorphological correlation. Possible indications for volumetry of gastrointestinal masses in the assessment of neoadjuvant therapeutic concepts are under scientific evaluation.


Lobular versus ductal breast neoplasms: the diagnostic utility of p120 catenin.
The distinction between lobular and ductal lesions of the breast is important in several circumstances. Diagnostic reproducibility of lobular versus ductal lesions, based on histology alone, is less than optimal. The proper distinction between atypical lobular hyperplasia, lobular carcinoma in situ and low-grade ductal carcinoma in situ is critical for patient management. Patients who have a core biopsy of invasive lobular carcinoma often have preoperative magnetic resonance imaging to prepare the surgeon for proper margin attainment. E-cadherin, a negative membrane marker for lobular neoplasia, is useful in the distinction of lobular versus ductal neoplasia, but as a negative marker, can be difficult to interpret in particularly challenging cases. In this study, we surveyed primary and metastatic ductal lesions (62) and lobular lesions (64) of the breast to determine if P120 catenin is useful in the diagnostic distinction between lobular and ductal neoplasia. Primary breast ductal and lobular preneoplastic and neoplastic lesions were immunostained with E-cadherin and P120ctn and independently classified as ductal or lobular lesions. In addition, a wide array of carcinomas of different types were surveyed with P120ctn in tissue microarrays to ascertain whether the cytoplasmic P120ctn immunostaining pattern observed in lobular neoplasia was unique. Accurate categorization of ductal versus lobular neoplasia in the breast with P120ctn immunostaining was effective in all cases. Separation of low-grade ductal carcinoma in situ from lobular neoplasia was efficient. Diagnostically, P120ctn was particularly useful in identifying early lesions of lobular neoplasia. Of the other tumors that may morphologically mimic lobular carcinoma, only the diffusely infiltrating variants of rectal and gastric carcinomas showed diffuse cytoplasmic P120ctn immunostaining. Caution should be exercised when examining tumors in metastatic sites with P120ctn, with the incorporation of an appropriate panel of immunostains.


Wheat (Triticum aestivum L. and T. turgidum L. ssp. durum) Kernel Hardness: I. Current View on the Role of Puroindolines and Polar Lipids.
Wheat hardness has major consequences for the entire wheat supply chain from breeders and millers over manufacturers to, finally, consumers of wheat-based products. Indeed, differences in hardness among Triticum aestivum L. or between T. aestivum L. and T. turgidum L. ssp. durum wheat cultivars determine not only their milling properties, but also the properties of flour or semolina endosperm particles, their preferential use in cereal-based applications, and the quality of the latter. Although the mechanism causing differences in wheat hardness has been subject of research more than once, it is still not completely understood. It is widely accepted that differences in wheat hardness originate from differences in the interaction between the starch granules and the endosperm protein matrix in the kernel. This interaction seems impacted by the presence of either puroindoline a and/or b, polar lipids on the starch granule surface, or by a combination of both. We focus here on wheat hardness and its relation to the presence of puroindolines and polar lipids. More in particular, the structure, properties, and genetics of puroindolines and their interactions with polar lipids are critically discussed as is their possible role in wheat hardness. We also address future research needs as well as the presence of puroindoline-type proteins in other cereals.


A definition of the Mycobacterium avium complex for taxonomical and clinical purposes, a review.
Nontuberculous mycobacteria, particularly the Mycobacterium avium complex (MAC) bacteria, are increasingly recognized as opportunistic pathogens of humans. As a result, studies on antibiotic treatment and taxonomy of the MAC are intensifying, but an updated definition of what constitutes the MAC, either for taxonomical studies or for clinical purposes, is lacking. On the basis of literature review and phylogenetic analyses, we propose to define the MAC as a grouping of slow-growing mycobacteria that show corresponding values in at least two of the following targets against either M. avium ATCC 25291T or Mycobacterium intracellulare ATCC 13950T: >99.4 % sequence identity for the full 16S rRNA gene, >98.7 % for the partial (5') 16S rRNA gene, >97.3 % for hsp65 and >94.4 % for rpoB region V. A >97.5 % value in concatenated analyses of >2500 bp that includes 16S rRNA, hsp65 and rpoB gene sequence data or ≥85 % average nucleotide identity to M. avium ATCC 25291T or M. intracellulare ATCC 13950T on basis of whole genome sequencing data is recommended. This molecular definition is based on the distances observed between the classical members of the MAC, M. avium and M. intracellulare. Applying this definition, the complex currently consists of 12 validly published species: Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium chimaera, Mycobacterium colombiense, Mycobacterium arosiense, Mycobacterium vulneris, Mycobacterium bouchedurhonense, Mycobacterium timonense, Mycobacterium marseillense, Mycobacterium yongonense, Mycobacterium paraintracellulare and Mycobacterium lepraemurium.


Choroideremia. A clinical and genetic study of 84 Finnish patients and 126 female carriers.
The aim of this work was to identify the choroideremia families in northern Finland, form an impression of the incidence of the disease in Finland, construct a picture of its clinical progression and gather new information on relevant genetic questions. A total of 111 choroideremia patients and 188 carriers were traced, members of four families from northern Finland and one from the Savo district. Ophthalmological confirmation was obtained for 84 choroideremia cases and 126 carriers. The largest of the families from northern Finland contained 80 cases of the disease and 146 carriers in eight generations among a total of more than 3000 descendants from one ancestral mother. The clinical picture for choroideremia proved to be more variable than could have been supposed from the literature, including cases of patients under 30 years of age who were already virtually blind and of patients of over 50 who were subjectively symptom-free. Only 7 out of 105 carriers could be shown anamnestically to have had subjective symptoms, but surprisingly, as many as 21 out of 52 carriers examined had changes in the visual field and 13 out of 40 examined showed deterioration in dark adaptation. One carrier was seen to undergo an obvious decline in dark adaptation during a three-year observation period. One indirect indication of the progression of fundus changes in choroideremia carriers was obtained from the fact that these changes, and also alterations in visual field and dark adaptation, were greater in the older carriers. A progression could also be detected by fundus photography in six instances, although the changes involved were fairly mild ones. Considerable variety was noted in the fundus findings for the choroideremia carriers, there being some 80-year-old subjects with quite minor changes and some 20-year-olds with obvious, extensive changes. Practical visual acuity remained normal throughout life in the majority of the carriers, however. Diagnosis within the known choroideremia families was fairly difficult, especially at the early stages in the survey, and even later on a few cases aged up to ten years produced diagnostic problems. Quite often diagnosis was easy, however, and choroideremic fundus changes were even identified in two boys aged 3 and 8 months. No other diseases could be shown to be associated with choroideremia, and the occurrence of dominantly inherited olivopontocerebellar atrophy alongside choroideremia in one branch of a family may be regarded as a coincidence.(ABSTRACT TRUNCATED AT 400 WORDS)


Risk factors for retinopathy of prematurity in six neonatal intensive care units in Beijing, China.
Guidelines on oxygenation policies and on the prevention and treatment of retinopathy of prematurity (ROP) were issued by the Chinese Medical Association in 2004. This study was undertaken to determine the incidence of and risk factors for ROP among preterm infants in Beijing, the capital of China, after implementation of the guidelines. Neonates with birth weights (BW) < or = 2000 g or gestational age (GA) < or = 34 weeks admitted to the six largest neonatal intensive care units in Beijing during 2005 were enrolled. Ophthalmological examinations started 3-4 weeks after birth, and ROP was classified using the revised International Classification. Maternal and perinatal risk factors for type 1 ROP were analysed. Retinopathy of prematurity was detected in 10.8% of 639 neonates who had complete eye examinations, 23 of whom (3.6%) developed type 1 ROP and were treated. The rate of ROP needing treatment has not declined since 2002. Logistic regression analysis indicated that low BW, apnoea > 20 s, anaemia, hypoxic-ischaemic encephalopathy and placenta abruption were significantly associated with type 1 ROP. In Beijing, rates of ROP needing treatment are high, and affected babies are more mature than in NICUs in high-income countries. More needs to be done to prevent ROP through improved neonatal care.


Direct identification of a peptide binding region in the opioid receptor-like 1 receptor by photoaffinity labeling with [Bpa(10),Tyr(14)]nociceptin.
The heptadecapeptide nociceptin, also known as orphanin FQ, is the endogenous agonist of the opioid receptor-like 1 (ORL1) G protein-coupled receptor. An affinity labeling approach has been implemented to probe the interactions of the neuropeptide with the receptor using the photolabile nociceptin derivative, [p-benzoyl-l-Phe(10),Tyr(14)]nociceptin ([Bpa(10),Tyr(14)]noc). In recombinant Chinese hamster ovary cells expressing the human ORL1 receptor, [Bpa(10),Tyr(14)]noc binds the receptor with high affinity (K(i) approximately 0.7 nm) and is as potent as nociceptin in the inhibition of forskolin-induced cAMP synthesis (EC(50) approximately 0.5 nm). UV irradiation at 365 nm of the complex formed by the ORL1 receptor and radioiodinated [Bpa(10),Tyr(14)]noc results in the irreversible labeling of a glycoprotein of approximately 65 kDa, determined by SDS-polyacrylamide gel electrophoresis. Complete digestion of the partially purified 65-kDa complex with kallikrein generates a single labeled fragment (approximately 6.5 kDa) that is readily cleaved by endoproteinase Glu-C to yield a labeled fragment of approximately 3.2 kDa. Kallikrein treatment of the photoaffinity cross-linked Glu(295) --> Asp mutant receptor also yields a single labeled fragment of approximately 6.5 kDa but is resistant to further cleavage by endoproteinase Glu-C. Based upon the expected proteolytic fingerprint of the labeled receptor, the photoreactive region can be identified as ORL1-(296-302; residues Thr-Ala-Val-Ala-Ile-Leu-Arg) spanning the C terminus of extracellular loop 3 and the N terminus of transmembrane helix VII. Molecular modeling of the ORL1 receptor complex with [Bpa(10)]noc suggests that reaction of the Bpa carbonyl group may occur with the side chain of Ile(300) within the experimentally identified photoreactive region.


A biomechanical and spectroscopic study of bone from rats with selenium deficiency and toxicity.
Selenium, being an essential mineral in the mammalian diet, is important in providing protection against oxidative damage. Numerous in vitro studies of selenium compounds reveal a very high correlation between catalytic activity of selenium compounds and toxicity. The present study was designed to investigate the effects of dietary selenium on the biomechanical properties of bone. New born rats of both sexes were fed with either a control, or a selenium- and vitamin E-deficient, or a selenium-excess and vitamin E-adequate diet. We obtained the stiffness (modulus of elasticity) of bones (femur and tibia) by tensile test for all groups considered. Both the deficient and the excess groups have decreased biomechanical strength with respect to the control group. To support our biomechanical results for both experimental groups, X-ray diffraction analysis and FTIR spectroscopic study were performed on the femurs and tibiae. The X-ray diffraction analysis showed that the intensities of the peak observed at around 2theta degrees = 31.820, in the control femur and tibia are stronger than the intensities of the corresponding peak of two experimental groups. In FTIR spectroscopy, the disappearance and/or reduction of the intensities of some carbonate bands in the two experimental groups indicate that there is a decrease in crystallinity and mineral contents which, together with X-ray diffraction analysis, correlate very well with the biomechanical data.