The molecular biology and evolution of feline immunodeficiency viruses of cougars.
Feline immunodeficiency virus (FIV) is a lentivirus that has been identified in many members of the family Felidae but domestic cats are the only FIV host in which infection results in disease. We studied FIVpco infection of cougars (Puma concolor) as a model for asymptomatic lentivirus infections to understand the mechanisms of host-virus coexistence. Several natural cougar populations were evaluated to determine if there are any consequences of FIVpco infection on cougar fecundity, survival, or susceptibility to other infections. We have sequenced full-length viral genomes and conducted a detailed analysis of viral molecular evolution on these sequences and on genome fragments of serially sampled animals to determine the evolutionary forces experienced by this virus in cougars. In addition, we have evaluated the molecular genetics of FIVpco in a new host, domestic cats, to determine the evolutionary consequences to a host-adapted virus associated with cross-species infection. Our results indicate that there are no significant differences in survival, fecundity or susceptibility to other infections between FIVpco-infected and uninfected cougars. The molecular evolution of FIVpco is characterized by a slower evolutionary rate and an absence of positive selection, but also by proviral and plasma viral loads comparable to those of epidemic lentiviruses such as HIV-1 or FIVfca. Evolutionary and recombination rates and selection profiles change significantly when FIVpco replicates in a new host.


Developing a scoring method for evaluating dietary methodology in reviews of epidemiologic studies.
We examined the quality of dietary assessment used by studies of prostate cancer and dietary fat in an attempt to explain the heterogeneity of their relative risk (RR) estimates. We reviewed the dietary assessment of 39 studies published in English that reported RRs for the association between prostate cancer and dietary fat intake derived from food frequency questionnaires (FFQs). We scored studies based on several objective measures of quality dietary assessment. Studies received no points for characteristics with unclear information. Studies scored 2 points for interviewer-completed FFQs, along with 2 points for quantitative assessments. They were scored 4 points for FFQs with more than 150 items, with an additional point for pretesting and 2 points for validated FFQs. Studies were given 1 point for describing each of the following characteristics: specifying the nutrient database used to convert foods to grams of fat, specifying quality control, attempting to measure dietary intake prior to diagnosis (recalled dietary period), and reporting the time needed to complete the FFQ. We then ranked studies based on their overall score: "high" for a score of 7 or greater out of 15 and "low" for lower scores. Two of the 39 studies that used quantitative methods other than a FFQ were excluded. Of the remaining 37 studies reviewed that used FFQs, only 16 were judged to have a high quality assessment of dietary fat. This review highlights the inconsistency of FFQ used in epidemiologic studies of dietary fat. Such variations in dietary measurement may be reflected in the variation in the magnitude of RRs reported for prostate cancer and dietary fat. The problems identified here include insufficient reporting of the details of dietary assessment, in addition to use of questionnaires with only a few food items to estimate a subject's dietary fat intake. It is imperative that journals include experts in the field of nutrition as reviewers of epidemiologic papers describing diet.


Aqueous sodium hydroxide (NaOH) solutions at high pressure and temperature: insights from in situ Raman spectroscopy and ab initio molecular dynamics simulations.
Hydrothermal diamond anvil cell experiments in combination with Raman spectroscopy and first principles molecular dynamics simulations were performed to investigate the structure and dynamics of aqueous NaOH solutions for temperatures up to 700 °C, pressures up to 850 MPa and two different solute concentrations. The significant changes observed in the O-H stretching region of the Raman spectra between ambient and supercritical conditions are explained by both dynamic effects and structural differences. Especially important are a Grotthuss-like proton transport process and the decreasing network connectivity of the water molecules with increasing temperature. The observed transfer of Raman intensity towards lower wavenumbers by the proton transfer affects a wide range of frequencies and must be considered in the interpretation of Raman spectra of highly basic solutions. We suggest a deconvolution of the spectra using a model with four Gaussian functions, which are assigned to the molecular H2O and OH- vibrations, and one asymmetric exponentially modified Gaussian (EMG) function, which is assigned to [HO(H2O)n]- vibrations.


Germline Features Associated with Immune Infiltration in Solid Tumors.
The immune composition of the tumor microenvironment influences response and resistance to immunotherapies. While numerous studies have identified somatic correlates of immune infiltration, germline features that associate with immune infiltrates in cancers remain incompletely characterized. We analyze seven million autosomal germline variants in the TCGA cohort and test for association with established immune-related phenotypes that describe the tumor immune microenvironment. We identify one SNP associated with the amount of infiltrating follicular helper T cells; 23 candidate genes, some of which are involved in cytokine-mediated signaling and others containing cancer-risk SNPs; and networks with genes that are part of the DNA repair and transcription elongation pathways. In addition, we find a positive association between polygenic risk for rheumatoid arthritis and amount of infiltrating CD8+ T cells. Overall, we identify multiple germline genetic features associated with tumor-immune phenotypes and develop a framework for probing inherited features that contribute to differences in immune infiltration.


Marihuana inhibits dihydrotestosterone binding to the androgen receptor.
Marihuana and its constitutents delta 9-tetrahydrocannabinol (delta 9-THC) and cannabinol (CBN) were tested for their ability to interact with the androgen receptor in rat prostate cytosol. Smoked marihuana condensate, delta 9-THC, and CBN inhibit specific binding of dihydrotestosterone (DHT) to the androgen receptor with a dissociatin constant of the inhibitors (Li) of 2.1-5.8 X 10(-7)M. in addition, other metabolites of delta 9-THC were also androgen antagonists. This data suggests that the anti-androgenic effects associated with marihuana use results, at least in part, from inhibition of androgen action at the receptor level.


[A tribute to professor Héctor Croxatto Rezzio: setting up science in Chile. "Nature is always more amazing than our imagination can conceive"].
Héctor R. Croxatto, M.D., has recently celebrated his 90th anniversary, fully active in scientific research and in other academic activities. Along his productive life, he has contributed with original observations on the role of certain peptides in the pathogenesis of arterial hypertension. Furthermore, he has been a leader in the development of biological research in Chile and the mentor or teacher of several prominent disciples who have had, or are having, their own brilliant careers in medicine and in science in Chile and other Latinamerican countries. Although most of his scientific productivity has been published in top journals in the international field, several of his papers have been published in Revista Médica de Chile. In this issue, the journal pays a tribute to this outstanding scholar, his exceptional personality and fruitful academic career. The tribute includes this Editorial, a Letter to the Editor and its reply, and his most recent manuscript, all testifying the exceptional virtues of a great man.


Degradation of emerging organic pollutants in wastewater effluents by electrochemical photocatalysis on nanostructured TiO2 meshes.
An immobilized photoactive TiO2 coating grown directly on titanium meshes was successfully exploited for the electrochemical photocatalytic degradation of carbamazepine in real secondary wastewater effluent. The catalyst was prepared by Plasma Electrolytic Oxidation and during the photocatalytic water treatment an electrical polarization (bias) was applied to the catalyst. The investigated process was compared with the conventional one employing suspended TiO2 powder (Degussa P25). Results showed that carbamazepine degradation rate follows the order UV/supported TiO2+bias ≈ UV/TiO2 Degussa P25 > UV/supported TiO2 > UV. The investigation also included the identification of other micropollutants and degradation products. This allowed the detection of 201 compounds present in the secondary wastewater effluent employed for the photocatalysis tests, 51 of them also successfully associated to compounds of emerging concern (CECs), and 194 to transformation products (TPs). The degradation of detected compounds followed first-order kinetics and the mean kinetic constant values of the 51 CECs resulted to be 0.048, 0.035 and 0.043 min-1 for the TiO2+Bias + UV, TiO2+UV and UV, respectively. As for TPs, results showed that the TiO2+Bias + UV treatment is much more efficient than both TiO2+UV and UV in minimizing the intensity of the organics in the real wastewater. Such a better performance was more pronounced at higher reaction time reaching 60% reduction of mean peak area of TPs at 90 min of reaction. Among the detected TPs also compounds belonging to known carbamazepine TPs were found. This allowed to propose a degradation pathway of carbamazepine. The supported catalyst was positively tested for 15 cycles demonstrating that it has the potential to be used in real wastewater tertiary steps aimed at removing CECs.


Cascade catalysis in membranes with enzyme immobilization for multi-enzymatic conversion of CO2 to methanol.
Facile co-immobilization of enzymes is highly desirable for bioconversion methods involving multi-enzymatic cascade reactions. Here we show for the first time that three enzymes can be immobilized in flat-sheet polymeric membranes simultaneously or separately by simple pressure-driven filtration (i.e. by directing membrane fouling formation), without any addition of organic solvent. Such co-immobilization and sequential immobilization systems were examined for the production of methanol from CO2 with formate dehydrogenase (FDH), formaldehyde dehydrogenase (FaldDH) and alcohol dehydrogenase (ADH). Enzyme activity was fully retained by this non-covalent immobilization strategy. The two immobilization systems had similar catalytic efficiencies because the second reaction (formic acid→formaldehyde) catalyzed by FaldDH was found to be the cascade bottleneck (a threshold substrate concentration was required). Moreover, the trade-off between the mitigation of product inhibition and low substrate concentration for the adjacent enzymes probably made the co-immobilization meaningless. Thus, sequential immobilization could be used for multi-enzymatic cascade reactions, as it allowed the operational conditions for each single step to be optimized, not only during the enzyme immobilization but also during the reaction process, and the pressure-driven mass transfer (flow-through mode) could overcome the diffusion resistance between enzymes. This study not only offers a green and facile immobilization method for multi-enzymatic cascade systems, but also reveals the reaction bottleneck and provides possible solutions for the bioconversion of CO2 to methanol.


REPLICATION OF THE RNA OF BACTERIOPHAGE R17.
Escherichia coli cells were irradiated with ultraviolet light to stop ribosomal RNA synthesis. After infection of such cells with the single-stranded RNA phage R17, the RNA most rapidly labeled by a pulse of tritiated uridine sedimented in a broad band in the 16S region of sucrose gradients. The effect of ribonuclease on this material and its behavior during a "chase" period in nonradioactive medium suggest that it consists of a core of double-stranded RNA, one strand of which-the viral strand-is continually displaced by a growing strand forming single-stranded tails and ultimately free 27S viral RNA.


A case of congenital cataracts with hypotrichosis caused by compound heterozygous variants in the LSS gene.
Patients with biallelic variants in the lanosterol synthase (LSS) gene has been reported to exhibit phenotypes as follows: non-syndromic form of hypotrichosis, congenital cataracts, and alopecia with intellectual disability or growth retardation. However, genotype-phenotype correlations in the LSS gene are still not completely clear. In this study, we reported a Chinese girl who had congenital cataracts with hypotrichosis. The trio exome sequencing was performed to elucidate the genetic cause of the patient. We identified compound heterozygous variants (c.296G>A, p.G99D and c.1025T>G, p.I342S) in the LSS gene. Both variants altered the amino acid coding at highly conserved amino acid residues and were predicted to be deleterious using prediction software. Our report expands the spectrum of variants in the LSS gene and will be helpful for genotype-phenotype correlations study.


Behavioral aspects of Angelman syndrome: a case control study.
Angelman syndrome (AS) is a rare congenital disorder characterized by impairments in intellectual, neurological and motor functioning and a postulated behavioral profile. This study compared behavioral characteristics of 62 individuals with genetically confirmed AS and 29 individuals with presumed AS from clinical features, with a control group of young persons with intellectual disability (ID) derived from an Australian epidemiological register. Twelve behavioral items from the developmental behavior checklist (DBC) were used for this comparison. The groups were matched for chronological age, gender, and level of ID. In the AS group, significant differences were found for 10 behaviors, with poor attention span and impulsivity being less common, and overactivity/restlessness, chewing or mouthing objects, eating non-food items, gorging food, food fads, fascination for water, hand flapping and sleep disturbance being more common. Interestingly, there was no difference in prevalence of unprovoked laughter. Comparison of the results of the genetically confirmed with the genetically unconfirmed AS cases showed no significant differences between individual behavior prevalence. These findings show that a "behavioral phenotype" of AS can be distinguished from others of similar level of ID, but it is different from that hitherto published. Abnormal food related behaviors, hyperactivity, fascination for water, hand flapping, and sleep disturbance should be included in a "behavioral phenotype" for AS. Apart from hyperactivity, "ADHD-type" behaviors are not more characteristic of AS than in ID generally. Therefore, the Consensus Criteria for the diagnosis of AS need to be reviewed.


The association between a single bout of moderate physical activity and executive function in young adults with Down syndrome: a preliminary study.
This study was aimed at investigating the impact of a single exercise intervention on executive function in young adults with Down syndrome (DS). Considering the relations among executive function, physical and mental health and early onset of Alzheimer's disease in this population, we tested three components of executive function (e.g. choice-response time, attention shifting and inhibition) that have been shown to be impaired in previous studies. Ten persons with DS were assigned to an exercise group, who walked on a treadmill for 20 min at moderate intensity and ten additional persons with DS were assigned to an attentional control group, who watched a video. Measures of executive function were tested pre and post interventions. These results showed non-significant improvements in choice-response time (P = 0.32) and attention shifting (P = 0.13) but a statistically significant improvement in inhibition (P = 0.03) after the exercise intervention. Given the improved inhibition ability, exercise may be an effective intervention, even in a signal session. However, only a few studies have focused on this topic. Based on theoretical models linking exercise to executive function, we proposed that exercise may increase arousal status or enhance neural transmission. Hence, future work is needed to examine the exact mechanism in the relationship between exercise and executive function for individuals with DS.


CT Scans in Young People in Great Britain: Temporal and Descriptive Patterns, 1993-2002.
Background. Although using computed tomography (CT) can be greatly beneficial, the associated relatively high radiation doses have led to growing concerns in relation to potential associations with risk of future cancer. Very little has been published regarding the trends of CT use in young people. Therefore, our objective was to assess temporal and other patterns in CT usage among patients aged under 22 years in Great Britain from 1993 to 2002. Methods. Electronic data were obtained from the Radiology Information Systems of 81 hospital trusts within Great Britain. All included patients were aged under 22 years and examined using CT between 1993 and 2002, with accessible radiology records. Results. The number of CT examinations doubled over the study period. While increases in numbers of recorded examinations were seen across all age groups, the greatest increases were in the older patients, most notably those aged 15-19 years of age. Sixty percent of CT examinations were of the head, with the percentages varying with calendar year and patient age. Conclusions. In contrast to previous data from the North of England, the doubling of CT use was not accompanied by an increase in numbers of multiple examinations to the same individual.


The pore of plant K(+) channels is involved in voltage and pH sensing: domain-swapping between different K(+) channel alpha-subunits.
Plant K(+) uptake channel types differ with respect to their voltage, Ca(2)+, and pH dependence. Here, we constructed recombinant chimeric channels between KST1, a member of the inward-rectifying, acid-activated KAT1 family, and AKT3, a member of the weakly voltage-dependent, proton-blocked AKT2/3 family. The homologous pore regions of AKT3 (amino acids 216 to 287) and KST1 (amino acids 217 to 289) have been exchanged to generate the two chimeric channels AKT3/(p)KST1 and KST1/(p)AKT3. In contrast to AKT3 wild-type channels, AKT3/(p)KST1 revealed a strong inward rectification reminiscent of that of KST1. Correspondingly, the substitution of the KST1 by the AKT3 pore led to less pronounced rectification properties of KST1/(p)AKT3 compared with wild-type KST1. Besides the voltage dependence, the interaction between the chimera and extracellular H(+) and Ca(2)+ resembled the properties of the inserted rather than the respective wild-type pore. Whereas AKT3/(p)KST1 was acid activated and Ca(2)+ insensitive, extracellular protons and Ca(2)+ inhibited KST1/(p)AKT3. The regulation of the chimeric channels by cytoplasmic protons followed the respective wild-type backbone of the chimeric channels, indicating that the intracellular pH sensor is located outside the P domain. We thus conclude that essential elements for external pH and Ca(2)+ regulation and for the rectification of voltage-dependent K(+) uptake channels are located within the channel pore.


Crucial Role of FABP3 in αSyn-Induced Reduction of Septal GABAergic Neurons and Cognitive Decline in Mice.
In synucleinopathies, while motor symptoms are thought to be attributed to the accumulation of misfolded α-synuclein (αSyn) in nigral dopaminergic neurons, it remains to be elucidated how cognitive decline arises. Here, we investigated the effects of distinct αSyn strains on cognition and the related neuropathology in the medial septum/diagonal band (MS/DB), a key region for cognitive processing. Bilateral injection of αSyn fibrils into the dorsal striatum potently impaired cognition in mice. The cognitive decline was accompanied by accumulation of phosphorylated αSyn at Ser129 and reduction of gamma-aminobutyric acid (GABA)-ergic but not cholinergic neurons in the MS/DB. Since we have demonstrated that fatty acid-binding protein 3 (FABP3) is critical for αSyn neurotoxicity in nigral dopaminergic neurons, we investigated whether FABP3 also participates in αSyn pathology in the MS/DB and cognitive decline. FABP3 was highly expressed in GABAergic but rarely in cholinergic neurons in the MS/DB. Notably, Fabp3 deletion antagonized the accumulation of phosphorylated αSyn, decrease in GABAergic neurons, and cognitive impairment caused by αSyn fibrils. Overall, the present study indicates that FABP3 mediates αSyn neurotoxicity in septal GABAergic neurons and the resultant cognitive impairment, and that FABP3 in this subpopulation could be a therapeutic target for dementia in synucleinopathies.


Exchange of Gramicidin between Lipid Bilayers: Implications for the Mechanism of Channel Formation.
The canonical mechanism of gramicidin (gA) channel formation is transmembrane dimerization of nonconducting subunits that reside in opposite bilayer leaflets. The channels do not open and close; they appear and disappear due to subunit association and dissociation. Many different types of experiments support this monomer ↔ dimer mechanism. Recently, however, this mechanism was challenged, based on experiments with lipid vesicle-incorporated gA under conditions where vesicle fusion could be controlled. In these experiments, sustained channel activity was observed long after fusion had been terminated, which led to the proposal that gA single-channel current transitions result from closed-open transitions in long-lived bilayer-spanning dimers. This proposal is at odds with 40 years of experiments, but involves the key assumption that gA monomers do not exchange between bilayers. We tested the possibility of peptide exchange between bilayers using three different types of experiments. First, we demonstrated the exchange of gA between 1,2-dierucoyl-sn-glycero-3-phosphocholine (DC22:1PC) or 1,2-dioleoyl-sn-glycero-3-phosphocholine (DC18:1PC) lipid vesicles using a fluorescence assay for gA channel activity. Second, we added gA-free DC22:1PC vesicles to both sides of planar DC18:1PC bilayers preincubated with gA, which reduced channel activity up to 10-fold. Third, we added gA-containing DC22:1PC vesicles to one or both sides of DC18:1PC planar bilayers, which produced much higher channel activity when the gA-containing vesicles were added to both sides of the bilayer, as compared to one side only. All three types of experiments show that gA subunits can exchange between lipid bilayers. The exchange of subunits between bilayers thus is firmly established, which becomes a crucial consideration with respect to the mechanism of channel formation.


A comparison of quality of life between older adults living in high and low altitude areas.
High altitude is known to have a significant impact on human physiology and health, therefore, understanding its relationship with quality of life is an important research area. This study compared the quality of life (QOL) in older adults living in high and low altitude areas, and examined the independent correlates of QOL in those living in a high altitude area. Older adults living in three public nursing homes in Xining (high altitude area) and one public nursing home in Guangzhou (low altitude area) were recruited. The WHOQOL-BREF was used to measure the QOL. 644 older adults (male: 39.1%) were included, with 207 living in high altitude and 437 living in low altitude areas. After controlling for the covariates, older adults living in the high altitude area had higher QOL in terms of physical (P = 0.035) and social domains (P = 0.002), but had lower QOL in psychological (P = 0.009) domain compared to their counterparts living in the low altitude area. For older adults living in the high altitude area, smoking status was associated with higher social QOL (P = 0.021), good financial status was associated with higher physical QOL (P = 0.035), and fair or good health status was associated with higher physical (p < 0.001) and psychological QOL (P = 0.046), while more severe depressive symptoms were associated with lower QOL. Appropriate interventions and support to improve depressive symptoms and both financial and health status should be developed for older adults living in high altitude areas to improve their QOL.


Evidence that both exosites on thrombin participate in its high affinity interaction with fibrin.
Exosite 1 on thrombin mediates low affinity binding to sites on the NH2 termini of the alpha- and beta-chains of fibrin. A subpopulation of fibrin molecules (gammaA/gamma'-fibrin) has an alternate COOH terminus of the normal gamma-chain (gammaA/gammaA-fibrin) that binds thrombin with high affinity. To determine the roles of exosites 1 and 2 in the high affinity interaction of thrombin with gammaA/gamma'-fibrin, binding studies were done with thrombin variants and exosite 1- or 2-directed ligands. alpha-Thrombin bound gammaA/gamma'-fibrin via high and low affinity binding sites. A peptide analog of the COOH terminus of the gamma'-chain that binds alpha-thrombin via exosite 2 blocked the high affinity binding of alpha-thrombin to gammaA/gamma'-fibrin, suggesting that the interaction of alpha-thrombin with the gamma'-chain is exosite 2-mediated. In support of this concept, (a) gamma-thrombin, which lacks a functional exosite 1, bound to gammaA/gamma'-fibrin, but not to gammaA/gammaA-fibrin; (b) thrombin R93A/R97A/R101A, an exosite 2-defective variant, bound only to gammaA/gamma'-fibrin via low affinity sites; and (c) exosite 2-directed ligands reduced alpha-thrombin binding to gammaA/gamma'-fibrin. However, several lines of evidence indicate that exosite 1 contributes to the high affinity interaction of thrombin with gammaA/gamma'-fibrin. First, the affinity of gamma-thrombin for gammaA/gamma'-fibrin was lower than that of alpha-thrombin. Second, removal of a low affinity binding site on the beta-chain of gammaA/gamma'-fibrin reduced its affinity for alpha-thrombin. Third, exosite 1-directed ligands reduced alpha-thrombin binding to gammaA/gamma'-fibrin. Taken together, these data suggest that, although exosite 2 mediates the interaction of thrombin with the gamma'-chain of gammaA/gamma'-fibrin, simultaneous ligation of exosite 1 by low affinity binding sites is essential for the high affinity interaction of thrombin with gammaA/gamma'-fibrin.


[Results of femoro-popliteal arterial reconstructions depending on hemodynamic conditions].
The investigation included 205 patients. Reconstructive operations were fulfilled on major arteries of the femoro-popliteal segment (FPS) of lower extremities. An analysis of long-term results showed that the results of surgical treatment of lesions of FPS in obliterating atherosclerosis of the lower extremity vessels depended on the level of the peripheral vascular resistance in the lower extremity arteries, and the formation of the arteriovenous shunt in reconstructions of the FPS allowed considerable improvement of results of surgical treatment of patients with high peripheral vascular resistance.


Suicide in Scottish prisons.
A consecutive series of prison suicides in Scotland between 1970 and 1982 was examined. The initial phase of imprisonment was found to be the most vulnerable stage of confinement, with almost two-thirds of the deaths having taken place within the first month. A disproportionately high number were found to be on remand. A history of some form of psychiatric involvement was common, and a third of the group had received previous psychiatric inpatient treatment. In contrast to results from general population studies, there was little evidence to link suicide with a history of depressive illness. A record of problems with either alcohol or drug dependence was found in almost half the cases. The importance of general measures designed to reduce stress and promote coping mechanisms is emphasised.


[Evaluation of operator's quality using test "Manometers" implemented with the use of hard- and software Neurolab-2000M].
Purpose of the work was to correlate the quality parameters (speed and accuracy) of test Manometers implementation with the use of hard- and software Neurolab-2000M. The operator's quality parameters were latent reaction time (LRT) as a speed criterion, number of errors as an accuracy criterion, and speed and accuracy dependent time of task presentation (TP). The parameters were related by evaluation of statistical significance of calculated linear correlation coefficients Correlations between TP and LRT as well as between LRT and the number of errors were neither unique nor significant. On the opposite, TP and the number of errors were in direct correlation on the level of proximity and high statistical significance. These results suggest that, like the number of errors, TP can serve as a criterion of test completion accuracy. Regarding LRT, the parameter is not suitable for evaluation of accuracy of the Manometers test completion.


Predictive Simulations with OpenSim Moco to Investigate the Interaction Between Human and Assistive Exoskeleton.
The purpose of this work was to investigate the interaction between human and lower limbs assistive exoskeleton under different levels of assistance, by using computational simulations. To this, a human-exoskeleton interaction model was used and three predictive simulations were carried out with the OpenSim Moco. The results proved that the increase in the level of robot assistance causes a reduction in human effort. In addition, it was possible to verify the RMS torque of both the robot and the human, as well as the muscle activations, for the different levels of assistance simulated. For future work, we intend to run predictive simulations with more complex movements, such as gait free and with obstacles, in addition to using models that can represent a human being with muscle weakness on one side of the body (hemiparesis).


A social and economic analysis of schistosomiasis: a conceptual framework and a research strategy.
This paper describes a broad conceptual framework for analyzing the social and economic aspects of schistosomiasis. In this framework, the household, in an attempt to improve its welfare, is assumed to make a variety of simultaneous decision based on a set of opportunities and constraints defined by its household resources and community environment, which are in turn influenced by public policies and programmes. The analytical gains from utilizing this broad framework in the study of the social and economic consequences of schistosomiasis, the transmission of the disease, and the demand for treatment from the perspective of more effective control of the disease are briefly noted. The paper also describes the research strategy being adopted by an ongoing research in the Philippines with attention to the intensive data requirements that the broad framework calls for.


Sonic hedgehog pathway activation is associated with cetuximab resistance and EPHB3 receptor induction in colorectal cancer.
A major problem of colorectal cancer (CRC) targeted therapies is relapse caused by drug resistance. In most cases of CRC, patients develop resistance to anticancer drugs. Cetuximab does not show many of the side effects of other anticancer drugs and improves the survival of patients with metastatic CRC. However, the molecular mechanism of cetuximab resistance is not fully understood. Methods: EPHB3-mediated cetuximab resistance was confirmed by in vitro western blotting, colony-forming assays, WST-1 colorimetric assay, and in vivo xenograft models (n = 7 per group). RNA-seq analysis and receptor tyrosine kinase assays were performed to identify the cetuximab resistance mechanism of EPHB3. All statistical tests were two-sided. Results: The expression of EFNB3, which upregulates the EPHB3 receptor, was shown to be increased via microarray analysis. When resistance to cetuximab was acquired, EPHB3 protein levels increased. Hedgehog signaling, cancer stemness, and epithelial-mesenchymal transition signaling proteins were also increased in the cetuximab-resistant human colon cancer cell line SW48R. Despite cells acquiring resistance to cetuximab, STAT3 was still responsive to EGF and cetuximab treatment. Moreover, inhibition of EPHB3 was associated with decreased STAT3 activity. Co-immunoprecipitation confirmed that EGFR and EPHB3 bind to each other and this binding increases upon resistance acquisition, suggesting that STAT3 is activated by the binding between EGFR and EPHB3. Protein levels of GLI-1, SOX2, and Vimentin, which are affected by STAT3, also increased. Similar results were obtained in samples from patients with CRC. Conclusion: EPHB3 expression is associated with anticancer drug resistance.


Sports-specialized intensive training and the risk of injury in young athletes: a clinical case-control study.
Data are lacking regarding the independent risk of injury related to intense single-sport training or growth rate in young athletes. To determine whether sports specialization, weekly training volumes, and growth rates are associated with increased risk for injury and serious overuse injury in young athletes. Case-control study; Level of evidence, 3. Injured athletes aged 7 to 18 years were recruited from 2 hospital-based sports medicine clinics and compared with healthy controls from affiliated primary care clinics undergoing sports physicals (2010-2013). Participants completed surveys reporting hours per week spent in organized sports, physical education class, and free play, as well as degree of sports specialization and Tanner stage. Heights and weights were measured. Injury details were obtained from athlete surveys and electronic medical records. Of 1214 athletes enrolled, 1190 (50.7% male) had data satisfactory for analysis. There were 822 injured participants (49.5% male; unique injuries, n = 846) and 368 uninjured participants (55% male). Injured athletes were older than uninjured athletes (14.1 ± 2.1 vs. 12.9 ± 2.6 years; P < .001) and reported more total hours of physical activity (19.6 ± 9.2 vs. 17.6 ± 8.9 h/wk; P < .001) and organized sports activity (11.2 ± 2.6 vs. 9.1 ± 6.3 h/wk; P < .01). After accounting for age and hours in sports activity spent per week, sports-specialized training was an independent risk for injury (odds ratio [OR], 1.27; 95% CI, 1.07-1.52; P < .01) and serious overuse injury (OR, 1.36; 95% CI, 1.08-1.72; P < .01). Young athletes participating in more hours of sports per week than number of age in years (OR, 2.07; 95% CI, 1.40-3.05; P < .001) or whose ratio of organized sports to free play time was >2:1 hours/week had increased odds of having a serious overuse injury (OR, 1.87; 95% CI, 1.26-2.76; P < .01). Growth rates were similar between injured and uninjured athletes (4.8 cm/y for both groups; P = .96). Injured young athletes were older and spent more hours per week in organized sports. There is an independent risk of injury and serious overuse injury in young athletes who specialize in a single sport. Growth rate was not related to injury risk. The study data provide guidance for clinicians counseling young athletes and their parents regarding injury risks associated with sports specialization.


Nanoparticulate glutathione peroxidase mimics based on selenocystine-pullulan conjugates.
We synthesized nanoparticulate glutathione peroxidase (GPx) mimics in which selenocystine (SeCyst) was conjugated to a hydrophilic linear polysaccharide, pullulan (Pul). The SeCyst ester-conjugated Pul derivatives (SeCyst-Pul) in phosphate buffer (pH 7) were treated with a sonicator to spontaneously form particulate materials. Dynamic light scattering measurements revealed that the SeCyst-Pul conjugates could form particulate materials with diameters between 100 and 300 nm. Distinctive endothermic peaks were observed for the SeCyst-Pul aggregate solutions based on a differential scanning calorimetric analysis. The tryptophan (Trp) fluorescence intensity of SeCyst benzyl ester-tryptophanyl-Pul (SeCyst-Bz-Trp-Pul) mostly decreased in comparison to those of the Trp-Pul (its precursor) and free Trp, which indicates that the Trp residues come close to each other during the aggregation of the conjugates. Formation of SeCyst-Pul aggregates could be induced by the hydrophobic interactions between the SeCyst esters and the amino acid residues on Pul. The GPx-like activity of SeCyst-Bz-Trp-Pul aggregates for the reduction of H2O2 was enhanced nearly 20-fold higher than that of free SeCyst. The double-reciprocal plots of the SeCyst-Bz-Trp-Pul aggregate-catalyzed reduction yielded parallel lines by varying the substrate concentrations, indicating a "ping-pong" mechanism that is similar to those of the natural GPxs. The enhanced GPx activity of the SeCyst-Bz-Trp-Pul aggregate was also supported by higher kinetic parameters, k(cat)/K(m) (GSH) and k(cat)/K(m) H2O2. Overall, the enhanced activity of the SeCyst-Bz-Trp-Pul aggregate would be attributed to a hydrophobic environment that was formed at the vicinity of the SeCyst.


Assessing the therapeutic potential of lab-made hepatocytes.
Hepatocyte transplantation has potential as a bridge or even alternative to whole-organ liver transplantation. Because donor livers are scarce, realizing this potential requires the development of alternative cell sources. To be therapeutically effective, surrogate hepatocytes must replicate the complex function and ability to proliferate of primary human hepatocytes. Ideally, they are also autologous to eliminate the need for immune suppression, which can have severe side effects and may not be sufficient to prevent rejection long term. In the past decade, several methods have been developed to generate hepatocytes from other readily and safely accessible somatic cells. These lab-made hepatocytes show promise in animal models of liver diseases, supporting the feasibility of autologous liver cell therapies. Here, we review recent preclinical studies exemplifying different types of lab-made hepatocytes that can potentially be used in autologous liver cell therapies. To define the therapeutic efficacy of current lab-made hepatocytes, we compare them to primary human hepatocytes, focusing on engraftment efficiency and posttransplant proliferation and function. In addition to summarizing published results, we discuss animal models and assays effective in assessing therapeutic efficacy. This analysis underscores the therapeutic potential of current lab-made hepatocytes, but also highlights deficiencies and uncertainties that need to be addressed in future studies aimed at developing liver cell therapies with lab-made hepatocytes. (Hepatology 2016;64:287-294).


Cross-sectional geometry predicts failure location in maize stalks.
Stalk lodging (breaking of agricultural plant stalks prior to harvest) is a multi-billion dollar a year problem. Stalk lodging occurs when high winds induce bending moments in the stalk which exceed the bending strength of the plant. Previous biomechanical models of plant stalks have investigated the effect of cross-sectional morphology on stalk lodging resistance (e.g., diameter and rind thickness). However, it is unclear if the location of stalk failure along the length of stem is determined by morphological or compositional factors. It is also unclear if the crops are structurally optimized, i.e., if the plants allocate structural biomass to create uniform and minimal bending stresses in the plant tissues. The purpose of this paper is twofold: (1) to investigate the relationship between bending stress and failure location of maize stalks, and (2) to investigate the potential of phenotyping for internode-level bending stresses to assess lodging resistance. 868 maize specimens representing 16 maize hybrids were successfully tested in bending to failure. Internode morphology was measured, and bending stresses were calculated. It was found that bending stress is highly and positively associated with failure location. A user-friendly computational tool is presented to help plant breeders in phenotyping for internode-level bending stress. Phenotyping for internode-level bending stresses could potentially be used to breed for more biomechanically optimal stalks that are resistant to stalk lodging. Internode-level bending stress plays a potentially critical role in the structural integrity of plant stems. Equations and tools provided herein enable researchers to account for this phenotype, which has the potential to increase the bending strength of plants without increasing overall structural biomass.


Effects of adding solid and molten chocolate on the physicochemical, antioxidant, microbiological, and sensory properties of ewe's milk cheese.
A novel dairy product, namely "chocolate cheese", was produced with two typical Sicilian food products: Pecorino cheese, processed from ewe's milk, and Modica chocolate. The cheese, manufactured with 0%, 5%, 10%, and 15% (w/w) solid or molten chocolate, was evaluated after 0, 2, 4, and 6 weeks of vacuum storage for its nutritional and health properties. The addition of chocolate reduced the pH, protein, fat, and ash; the addition of 5% or 10% molten chocolate reduced hardness (N/mm2 ). The addition of either solid or molten chocolate resulted in a slight increase (P < 0.1038) in the total polyphenol content, a higher oleic acid content, and less oxidative stability. The microbiological profile showed that the total mesophilic count and the number of mesophilic coccus lactic acid bacteria (LAB) were approximately equal (about 108 CFU/g) in all cheese. The survival of the microorganisms was affected by both the chocolate added and the storage time. Chocolate cheese stored for 6 weeks had less Enterobacteriaceae than control cheese, whereas yeasts were detected at higher cell densities in the former cheese. Filamentous fungi were undetectable in some cheese. Differences were also observed in the number of mesophilic rod LAB, which increased progressively over time in all cheese, and in Enterobacteriaceae, yeasts, and filamentous fungi, which decreased during storage. Descriptive and hedonic sensory tests and principal component analysis showed that fresh cheese and cheese stored for 2 weeks, including 5% molten chocolate, were the most preferred by evaluators. Based on these results, chocolate cheese has the potential to be appreciated in the market for its nutritional, health, and sensory properties. PRACTICAL APPLICATION: Chocolate cheese, made by combining two typical Sicilian foods, Pecorino cheese and Modica chocolate, is proposed as a novel dairy product. The highest sensory acceptance was obtained with the addition of 5% molten chocolate and storage for 2 weeks. Given its improved antioxidant properties, healthier fat, and sensory properties, chocolate cheese has the potential to be appreciated in the market, especially by young consumers.


Further studies on cynodont endocasts (Reptilia--Therapsida).
Endocranial casts of the cynodonts Andescynodon mendozensis, Probelesoden sp., Massetognathus pascualis and M. teruggii are studied qualitatively and quantitatively. The former two show large and flat olfactory bulbs, little or no representation of the olfactory peduncles, long and narrow cerebral region and laterally protruding flocculus at the cerebellar region, specially developed in Andescynodon. The specimens of Massetognathus show long, narrow and elevated olfactory bulbs, well represented olfactory peduncles, wide cerebral region (specially in M. pascuali) and pineal foramen (specially developed in M. teruggii). The encephalization quotients are in agreement with previous results on cynodont endocasts. One specimen of Andescynodon shows a range of 0.097-0.15 and Massetognathus pascuali 0.12-0.18.


Associated factors for Tracheostomy in adults with severe traumatic brain injury. Score proposal.
In patients with severe traumatic brain injury (TBI), there is a lack of consensus about the need and time to perform a tracheostomy. Nowadays, the decision is individualized to each case. It is considered that patients that will need a tracheostomy profit by performing it earlier. An observational and prospective study was performed. One hundred and twenty patients in a period of 18 months between 2016 and 2018, older than 18 years, with severe TBI at the first 24 h of trauma were analyzed. Clinical, biochemical, and radiological findings at admission were measured; patients were followed up until discharge. The statistical analysis was made with Student's t-test, χ2, and prevalence risk ratio. Ten associated factors were grouped according to the prevalence risk ratio. The principal factors were CRASH score, IMPACT score, SAPS II score, APACHE II score, age, revised trauma score, Glasgow Coma Scale, subdural hematoma, uni or bilateral abnormal pupil reactivity, and collapse of basal cisterns. We conclude that there are multiple factors associated with the need for tracheostomy in adult patients with severe TBI and it is possible to predict according to our findings from admission which patients will profit by this procedure.


[Laminar analysis of neuronal ultrastructure in the midbrain tectum of the steppe turtle].
A layer-to-layer study of the neuronal ultrastructure in the mesencephalic optic centre of T. horsfieldi has been made, the results being compared to those of similar experments with Emys orbicularis L. Neurons of the efferent type (layers IV, V), in contrast to short axon elements (layers, I, 22) are characterized by a greater amount of cytoplasm and by a variety of cytoplasmatic organelles. A few morphological types of neurons are distinguished: light, dark and neurons with a light nucleus and a dark cytoplasm. In two systems of horizontal neurons dark cells predominate to be assumed as being an independent morphofunctional neuronal type. In contrast to E. orbicularis, the neurons of T. horsfieldi show a large amount of axo-somatic synapses on the surface, and a deficiency in spiny outgrowths, both on the apical and basal dendrites.


Diagnostic reference levels in pediatric radiology in Austria.
Because of the higher sensitivity to radiation in children, optimization plays an essential role in pediatric radiology. Diagnostic reference levels are a helpful tool to optimize patient dose for standard radiographic procedures. A nationwide survey was conducted to determine doses and variation in average dose applied to children for standard X-ray examinations. Dose values for chest, skull and abdomen X-ray procedures were determined for newborns, 1, 5, 10 and 15 year olds. Third quartiles of patient doses are provided and compared to European, British and German reference values. Variation in average doses between hospitals (i.e., ratio of maximal to minimal applied average dose) for the same age and examination was found to be typically a factor of eight. Ratios of maximal to minimal applied doses to individual patients were found to be approximately a factor of six higher than variations of average doses between clinics. While average patient doses indicate the use of good radiological practice in general, variance in average doses applied by different hospitals indicate that audit of dose with respect to reference values will provide guidance for optimization. Such practice should result in lowering doses for pediatric examinations.


[Helicobacter pylori infection and gastric cancer in Mexico. A challenge for prevention and population control].
The International Agency for Research on Cancer (IARC) of the WHO has recognized a cause-effect relationship between Helicobacter pylori (Hp) infection and gastric cancer of such magnitude that the presence of this infection increases the risk of gastric cancer approximately four times. Gastric cancer is currently the second cause of mortality due to malignant neoplasms in Mexico City. This article explores the association between Hp infection and gastric cancer incidence through an epidemiological study including 109 gastric cancer patients and 177 hospital controls in Mexico City. The study estimates that, in the population studied, Hp infection was present in 87.2% of the cases, compared with 82.5% of the controls. The odds ratio of having gastric cancer if infected with Hp was 1.44 IC95% 0.7-2.8. In addition, it was calculated that with eradication of Hp infection in the general population, gastric cancer incidence would decrease by at least 26.6%. An improvement of the actual sanitary conditions along with the development of an effective vaccine for Hp infection and the existence of increasingly effective treatments to eradicate the bacteria are the necessary next step for populational prevention and control of gastric cancer.


Patient-centered connectivity-based prediction of tau pathology spread in Alzheimer's disease.
In Alzheimer's disease (AD), the Braak staging scheme suggests a stereotypical tau spreading pattern that does, however, not capture interindividual variability in tau deposition. This complicates the prediction of tau spreading, which may become critical for defining individualized tau-PET readouts in clinical trials. Since tau is assumed to spread throughout connected regions, we used functional connectivity to improve tau spreading predictions over Braak staging methods. We included two samples with longitudinal tau-PET from controls and AD patients. Cross-sectionally, we found connectivity of tau epicenters (i.e., regions with earliest tau) to predict estimated tau spreading sequences. Longitudinally, we found tau accumulation rates to correlate with connectivity strength to patient-specific tau epicenters. A connectivity-based, patient-centered tau spreading model improved the assessment of tau accumulation rates compared to Braak stage-specific readouts and reduced sample sizes by ~40% in simulated tau-targeting interventions. Thus, connectivity-based tau spreading models may show utility in clinical trials.


Reaching perinatal women online: the Healthy You, Healthy Baby website and app.
Overwhelming evidence reveals the close link between unwarranted weight gain among childbearing women and childhood adiposity. Yet current barriers limit the capacity of perinatal health care providers (PHCPs) to offer healthy lifestyle counselling. In response, today's Internet savvy women are turning to online resources to access health information, with the potential of revolutionising health services by enabling PHCPs to guide women to appropriate online resources. This paper presents the findings of a project designed to develop an online resource to promote healthy lifestyles during the perinatal period. The methodology involved focus groups and interviews with perinatal women and PHCPs to determine what online information was needed, in what form, and how best it should be presented. The outcome was the development of the Healthy You, Healthy Baby website and smartphone app. This clinically-endorsed, interactive online resource provides perinatal women with a personalised tool to track their weight, diet, physical activity, emotional wellbeing, and sleep patterns based on the developmental stage of their child with links to quality-assured information. One year since the launch of the online resource, data indicates it provides a low-cost intervention delivered across most geographic and socioeconomic strata without additional demands on health service staff.


Circumventing the stability-function trade-off in an engineered FN3 domain.
The favorable biophysical attributes of non-antibody scaffolds make them attractive alternatives to monoclonal antibodies. However, due to the well-known stability-function trade-off, these gains tend to be marginal after functional selection. A notable example is the fibronectin Type III (FN3) domain, FNfn10, which has been previously evolved to bind lysozyme with 1 pM affinity (FNfn10-α-lys), but suffers from poor thermodynamic and kinetic stability. To explore this stability-function compromise further, we grafted the lysozyme-binding loops from FNfn10-α-lys onto our previously engineered, ultra-stable FN3 scaffold, FN3con. The resulting variant (FN3con-α-lys) bound lysozyme with a markedly reduced affinity, but retained high levels of thermal stability. The crystal structure of FNfn10-α-lys in complex with lysozyme revealed unanticipated interactions at the protein-protein interface involving framework residues of FNfn10-α-lys, thus explaining the failure to transfer binding via loop grafting. Utilizing this structural information, we redesigned FN3con-α-lys and restored picomolar binding affinity to lysozyme, while maintaining thermodynamic stability (with a thermal melting temperature 2-fold higher than that of FNfn10-α-lys). FN3con therefore provides an exceptional window of stability to tolerate deleterious mutations, resulting in a substantial advantage for functional design. This study emphasizes the utility of consensus design for the generation of highly stable scaffolds for downstream protein engineering studies.


TXNL1-XRCC1 pathway regulates cisplatin-induced cell death and contributes to resistance in human gastric cancer.
Cisplatin is a cytotoxic platinum compound that triggers DNA crosslinking induced cell death, and is one of the reference drugs used in the treatment of several types of human cancers including gastric cancer. However, intrinsic or acquired drug resistance to cisplatin is very common, and leading to treatment failure. We have recently shown that reduced expression of base excision repair protein XRCC1 (X-ray repair cross complementing group1) in gastric cancerous tissues correlates with a significant survival benefit from adjuvant first-line platinum-based chemotherapy. In this study, we demonstrated the role of XRCC1 in repair of cisplatin-induced DNA lesions and acquired cisplatin resistance in gastric cancer by using cisplatin-sensitive gastric cancer cell lines BGC823 and the cisplatin-resistant gastric cancer cell lines BGC823/cis-diamminedichloridoplatinum(II) (DDP). Our results indicated that the protein expression of XRCC1 was significantly increased in cisplatin-resistant cells and independently contributed to cisplatin resistance. Irinotecan, another chemotherapeutic agent to induce DNA damaging used to treat patients with advanced gastric cancer that progressed on cisplatin, was found to inhibit the expression of XRCC1 effectively, and leading to an increase in the sensitivity of resistant cells to cisplatin. Our proteomic studies further identified a cofactor of 26S proteasome, the thioredoxin-like protein 1 (TXNL1) that downregulated XRCC1 in BGC823/DDP cells via the ubiquitin-proteasome pathway. In conclusion, the TXNL1-XRCC1 is a novel regulatory pathway that has an independent role in cisplatin resistance, indicating a putative drug target for reversing cisplatin resistance in gastric cancer.


In the Gray Zone in the Fragile X Gene: What are the Key Unanswered Clinical and Biological Questions?
Smaller expansions (41-54 CGG repeats) in the fragile X mental retardation 1 (FMR1) gene are termed "gray zone" alleles. Only recently has interest in these expansions increased due to reporting of phenotypes unique to gray zone carriers or similar to those seen in individuals with larger expansions. As minimal research has focused on gray zone expansions, this paper asks several questions related to this topic. These include the following: What is the definition of the gray zone? Is there a risk of developing neurological signs in these carriers? Are there secondary gene effects that impact gray zone alleles or a biologic advantage to carrying these repeats? How do we counsel patients with gray zone expansions? The answers to these questions will help to determine the significance of these expansions and provide needed information to the research community and clinicians.


Optimized MR imaging for polyacrylamide gel dosimetry.
Polyacrylamide gels are a powerful tool to measure radiation dose by quantifying the NMR T2 relaxation times of the irradiated gel. The exploitation of these radiation sensitive gels in clinical radiotherapy requires accurate mapping of T2 values. This paper describes the optimization strategy used to identify accurate and practical methods of measuring the range of T2 values typical of gel dosimeters (140-700 ms). The MR imaging techniques used to measure T2 values and the choice of image acquisition parameters are described. Four sequences are compared and the results are analysed in terms of accuracy, signal-to-noise ratio and acquisition time. A multiple spin echo sequence was found to yield the most accurate results (98.9%). Single spin echo sequences, such as Hahn spin echo and EPI spin echo, were found to measure gel T2 values with an accuracy of 90.1%. This paper reports the importance of careful selection and optimization of the MR imaging sequences for accurate and reliable polyacrylamide gel dosimetry.


Intradiscal therapy: a review of current treatment modalities.
A systematic review of the medical literature regarding current intradiscal therapeutic methods. To establish an understanding of the current intradiscal treatment options for the management of low back pain. Current treatment of intradiscal disease is rapidly evolving and as such should be a multidisciplinary effort that follows a logical, orderly algorithm. Minimally invasive techniques, namely, intradiscal electrothermal therapy (IDET), radiofrequency ablation (RFA), percutaneous endoscopic laser discectomy (PELD), and cryoablation have challenged the conventional surgical management of back pain. Thirty-eight research reports, published between 1986 and 2005, were systematically reviewed for disease classification, surgical intervention, and treatment outcomes (neurologic status, pain scores, and ambulation). The surgical literature on the management of intradiscal disease continues to be limited to large series with short clinical follow-ups. Arthrodesis continues to be the primary treatment modality in the majority of patients. Newer treatment options including IDET, RFA, PELD, and cryoablation have shown promising results with regards to symptomatic relief and early return to function. Low back pain is an extremely common and potentially debilitating problem. Adding biophysical methods to well-tested biomechanical and newly investigated biomolecular solutions allows for multiple avenues of therapeutic interventions. With future clinical and basic science studies regarding intradiscal therapies forthcoming, we may soon alter our current treatment algorithms for the management of discogenic back pain.


Forestomach epithelial receptor activation by rumen fluids from sheep given intraruminal infusions of volatile fatty acids.
Forty-four reticuloruminal epithelial receptors were tested with rumen fluids obtained from 12 sheep before they were intraruminally infused with 4.0M acetic acid (8 sheep) or 4.0M butyric acid (4 sheep; preinfusion rumen fluid) and with rumen fluids obtained at the onset of ruminal stasis (abolition rumen fluid). The preinfusion rumen fluids from the 8 acetic acid-infused sheep (mean pH, 6.55) contained 1.7 mM nondissociated volatile fatty acids (VFA)/L and excited none of the 25 receptors tested. Preinfusion rumen fluids from the 4 butyric acid-infused sheep (mean pH, 6.98) contained 0.3 mM nondissociated VFA/L and also did not evoke responses in any of the 19 receptors tested. Abolition rumen fluids from sheep treated with acetic acid excited 17 of the 25 receptors tested and contained 89.4 mM nondissociated VFA/L, of which nondissociated acetic acid comprised 85.0 mM/L. Abolition rumen fluids from sheep treated with butyric acid activated 14 of the 19 receptors tested and contained 61.1 mM nondissociated VFA/L, of which 38.7 mM/L was nondissociated butyric acid. Preinfusion rumen fluids whose pH values were adjusted to that of abolition rumen fluids with HCl contained nondissociated VFA levels ranging from 16.3 mM/L (acetic acid-treated sheep) to 20.6 mM/L (butyric acid-treated sheep) and elicited responses in 4 of 30 receptors tested. Preinfusion rumen fluids whose pH values were adjusted to the pH value of abolition rumen fluid with acetic acid contained 29.5 mM nondissociated VFA/L and excited 7 of 13 tested receptors.(ABSTRACT TRUNCATED AT 250 WORDS)


Solution structure and folding characteristics of the C-terminal SH3 domain of c-Crk-II.
Crk-II is a signaling adaptor protein that is involved in many cellular processes including apoptosis, proliferation, and differentiation. It has a modular domain architecture consisting of an Src homology 2 domain (SH2) followed by two Src homology 3 (SH3) domains. The structures and ligand-binding properties of the SH2 and the middle SH3 domains are well-characterized. Several studies suggest that the C-terminal SH3 domain plays an important regulatory role in the protein; however, no structural information is available on this domain, and relatively little is known about its binding partners. In the current work, we have solved the solution NMR structure of the C-terminal SH3 domain. The domain adopts the standard SH3 fold comprising a five-stranded beta barrel. In agreement with alignment and modeling studies, the structure indicates that the canonical-binding surface of the SH3 domain is unusually polar and suggests that this domain may not bind typical PXXP ligands or that it may bind them with reduced affinity. Thermodynamic and kinetic studies show that the domain folds in a reversible two-state manner and that the stability of the fold is similar to that observed for other SH3 domains. These studies offer some insight into the likely structural and thermodynamic consequences of point mutations in the cSH3 domain that are known to deregulate Crk-II function. Our results set the stage for a better understanding the role of the cSH3 domain in the context of the full-length protein.


Selaginella moellendorffii telomeres: conserved and unique features in an ancient land plant lineage.
Telomeres, the essential terminal regions of linear eukaryotic chromosomes, consist of G-rich DNA repeats bound by a plethora of associated proteins. While the general pathways of telomere maintenance are evolutionarily conserved, individual telomere complex components show remarkable variation between eukaryotic lineages and even within closely related species. The recent genome sequencing of the lycophyte Selaginella moellendorffii and the availability of an ever-increasing number of flowering plant genomes provides a unique opportunity to evaluate the molecular and functional evolution of telomere components from the early evolving non-seed plants to the more developmentally advanced angiosperms. Here we analyzed telomere sequence in S. moellendorffii and found it to consist of TTTAGGG repeats, typical of most plants. Telomere tracts in S. moellendorffii range from 1 to 5.5 kb, closely resembling Arabidopsis thaliana. We identified several S. moellendorffii genes encoding sequence homologs of proteins involved in telomere maintenance in other organisms, including CST complex components and the telomere-binding proteins, POT1 and the TRFL family. Notable sequence similarities and differences were uncovered among the telomere-related genes in some of the plant lineages. Taken together, the data indicate that comparative analysis of the telomere complex in early diverging land plants such as S. moellendorffii and green algae will yield important insights into the evolution of telomeres and their protein constituents.


[Effect of the inoculum on tobramycin biosynthesis].
The effect of the quality and quantity of the inoculum on the biosynthesis of tobramycin was studied. The medium with corn steep liquor and glycerol was found to be the optimal seed medium for the tobramycin-producing organism. It is required that the fermentation medium be supplemented with 2 per cent of the vegetative inoculum at the stationary growth phase or with an increased amount of the seed material at the exponential growth phase or at the early stage of the culture dying. Subculturing of the inoculum decreases the level of tobramycin production at the fermentation stage: when the culture of the 5th--6th generation is used, the activity of the fermentation broth decreases by 30--50 per cent.


Human plasma and human platelet-rich plasma as a substitute for fetal calf serum during long-term cultivation of mesenchymal dental pulp stem cells.
Our aims were to isolate and cultivate mesenchymal dental pulp stem cells (DPSC) in various media enriched with human blood components, and subsequently to investigate their basic biological properties. DPSC were cultivated in five different media based on α MEM containing different concentrations of human plasma (HP), platelet-rich plasma (PRP), or fetal calf serum (FCS). The DPSC biological properties were examined periodically. We cultivated DPSC in the various cultivation media over 15 population doublings except for the medium supplemented with 10% HP. Our results showed that DPSC cultivated in medium supplemented with 10% PRP showed the shortest average population doubling time (DT) (28.6 ± 4.6 hours), in contrast to DPSC cultivated in 10% HP which indicated the longest DT (156.2 ± 17.8 hours); hence this part of the experiment had been cancelled in the 6th passage. DPSC cultivated in media with 2% FCS+ITS (DT 47.3 ± 10.4 hours), 2% PRP (DT 40.1 ± 5.7 hours) and 2% HP (DT 49.0 ± 15.2 hours) showed almost the same proliferative activity. DPSC's viability in the 9th passage was over 90% except for the DPSC cultivated in the 10% HP media. We proved that human blood components are suitable substitution for FCS in cultivation media for long-term DPSC cultivation.


Eating disorders and body image disturbances among ballet dancers, gymnasium users and body builders.
Eating disorders are frequent among elite performers of certain sports or physical activities; however, little is known about non-professional performers. 113 female non-elite ballet dancers, 54 female gymnasium users, 44 male non-competitive body builders, 105 female controls and 30 male controls were evaluated using the Body Uneasiness Test, the State-Trait Anxiety Inventory, the Beck Depression Inventory, and the Eating Disorder Examination 12th edition (EDE-12). Non-elite ballet dancers reported the highest prevalence of eating disorders (anorexia nervosa 1.8%; bulimia nervosa 2.7%; eating disorders not otherwise specified 22.1%), followed by gymnasium users (anorexia nervosa 2.6%; eating disorders not otherwise specified 18%). Significant differences (p < 0.01) between athletes and their controls were found in the following parameters (median values): Beck Depression Inventory (female dancers 5.7, gymnasium users 6.1, female controls 2.8, body builders 1.6, and male controls 1.3), Body Uneasiness Test (female dancers 1.08, gymnasium users 0.62, female controls 0.54, body builders 0.35, and male controls 0.27), EDE total scores (female dancers 1.6, gymnasium users 1.7, female controls 1.0, body builders 1.0, and male controls 0.4), EDE - restraint subscale scores (female dancers 0.8, gymnasium users 1.6, female controls 0.0, body builders 0.8, and male controls 0.0), EDE--eating concern subscale scores (female dancers 0.4, gymnasium users 0.2, female controls 0.0, body builders 0.0, and male controls 0.0), EDE--weight concern subscale scores (female dancers 2.1, gymnasium users 2.1, female controls 1.6, body builders 1.4, and male controls 0.5), and EDE--shape concern subscale scores (female dancers 2.7, gymnasium users 2.8, female controls 2.0, body builders 2.1, and male controls 0.9). EDE scores were highly related to Body Uneasiness Test scores, especially in non-elite ballet dancers and in non-competitive body builders (p < 0.01). Non-professional performers of sports emphasising thinness or muscularity, such as ballet and body-building, show a high degree of body uneasiness and inappropriate eating attitudes and behaviours.


The nuclear pore complex protein Tpr is a common autoantigen in sera that demonstrate nuclear envelope staining by indirect immunofluorescence.
We studied the autoantigen targets of 75 human sera that had antibodies to the nuclear envelope (NE) as identified by indirect immunofluorescence (IIF) on HEp-2 cells. Several different IIF staining patterns could be identified when antibodies to different components of the nuclear membrane (NM) and nuclear pore complexes (NuPC) were identified: a smooth membrane pattern characteristic of antibodies to nuclear lamins, a punctate pattern typical of antibodies to the nuclear pore complex and more complex patterns that included antibodies to nuclear and cytoplasmic organelles. Western immunoblotting of isolated nuclear and NE proteins and immunoprecipitation of radiolabelled recombinant proteins prepared by using the full-length cDNAs of the Translocated promoter region (Tpr), gp210 and p62 were used to identify specific autoantibody targets. Fifty-two of the 75 (70%) sera bound to Tpr, 25 (33%) bound to lamins A, B or C, 15 (20%) reacted with gp210 and none reacted with p62. Sixteen (21%) did not react with any of the NE components tested in our assays. The clinical features of 37 patients with anti-NE showed that there were 34 females and three males with an age range of 16-88 years (mean 59 years). The most frequent clinical diagnosis (9/37 = 24%) was autoimmune liver disease (ALD; two with primary biliary cirrhosis), followed by seven (19%) with systemic lupus erythematosus (SLE), four (11%) with a motor and/or sensory neuropathy, three (8%) with anti-phospholipid syndrome (APS), two with systemic sclerosis (SSc), two with Sjögren's syndrome (SjS), and others with a variety of diagnoses. This report indicates that Tpr, a component of the NuPC, is a common target of human autoantibodies that react with the NE.


Detection of aberrantly methylated hMLH1 promoter DNA in the serum of patients with microsatellite unstable colon cancer.
Serological tumor markers have proven valuable in the care of individuals with cancer for the early detection of primary cancers, early detection of cancer relapse, monitoring the response of cancers to therapy, and as predictors of cancer prognosis. Recently, the aberrant hypermethylation of the hMLH1 promoter and its consequent transcriptional silencing has been shown to be a common event in the formation of sporadic microsatellite unstable colon cancer. The silencing of hMLH1 expression appears to be controlled by the hypermethylation of a specific region in the hMLH1 promoter. We developed a methylation-specific PCR assay that assesses this region of the hMLH1 promoter. We found that this assay is able to detect methylated hMLH1 promoter DNA in the serum of some patients with microsatellite unstable colon cancers. In a panel of sera from 19 colon cancer cases, 9 with hMLH1 promoter methylation in the tumor primary, the assay proved 33% sensitive and 100% specific. This assay offers a potential means for the serum-based detection and/or monitoring of microsatellite unstable colon cancers.


Pemetrexed plus gemcitabine as first-line chemotherapy for patients with peritoneal mesothelioma: final report of a phase II trial.
Pemetrexed in combination with cisplatin is approved for the treatment of pleural mesothelioma and is active in malignant peritoneal mesothelioma (MPeM). Pemetrexed and gemcitabine are synergistic in preclinical models, but the activity of this combination in MPeM is unknown. This clinical study assessed safety and efficacy of pemetrexed plus gemcitabine in chemotherapy-naïve patients with MPeM. Treatment consisted of gemcitabine 1,250 mg/m(2) on days 1 and 8, and pemetrexed 500 mg/m(2) on day 8, administered immediately before gemcitabine. Treatment was repeated every 21 days for six cycles or until disease progression. All patients received folic acid, vitamin B(12), and dexamethasone supplementation. End points included tumor response, toxicity, time to disease progression (TTPD), and overall survival (OS). Disease control rate (DCR) was also calculated. Twenty patients were enrolled between December 2002 and May 2004. The confirmed response rate was 15% (95% CI, 3.2% to 37.9%), with three patients experiencing a partial response. The DCR was 50% (95% CI, 27.2% to 72.8%). The most common grade 3 to 4 nonhematologic toxicities included fatigue (20%), constipation (10%), vomiting (10%), and dehydration (10%). Hematologic toxicities included grade 3 to 4 neutropenia (60%) and febrile neutropenia (10%). One patient death was attributed to treatment. Median TTPD and OS times were 10.4 months and 26.8 months, respectively. The combination of pemetrexed plus gemcitabine was active in patients with MPeM with a notably high incidence of neutropenia. Median TTPD and OS seem promising. This regimen may provide an alternative to standard therapies, especially for patients who cannot tolerate a platinum-based regimen.


Improved clinical utility in clavicle fracture decision-making with true orthogonal radiographs.
Operative treatment of displaced clavicle fractures has demonstrated lower rates of nonunion and symptomatic malunion, improved functional outcomes, and earlier return to activities versus nonoperative treatment. Surgical treatment may offer a reduction in the relative risk of nonunion and symptomatic malunion and an earlier return to work or sport. To present an easy and safe method of acquiring orthogonal views of the clavicle without any manipulation of the patient's upper extremity. Academic medical center and private clinic in the Midwestern United States; retrospective patient cohort. We reviewed records of patients with acute clavicle fractures between January 2010 and August 2011. Thirty-four patients were treated with sling immobilization and 52 patients were treated surgically with superior plate and screw internal fixation. Prior to the introduction of orthogonal views, 19 patients were treated nonoperatively and 22 treated with internal fixation. Addition of orthogonal views increased the number of patients treated surgically: In the same period, 15 patients were treated nonoperatively and 30 with internal fixation. Following surgery, patients were evaluated in the outpatient office at 2 and 6 weeks and 3 and 6 months. Nonoperatively treated patients were evaluated at 1, 2, and 4 weeks and 3 and 6 months postinjury. Standard use of orthogonal views has led to greater utilization of surgical treatment of clavicle fractures at our institution and improved both the understanding and treatment of these injuries.


Identifying the molecular basis of host-parasite coevolution: merging models and mechanisms.
Mathematical models of the coevolutionary process have uncovered consequences of host-parasite interactions that go well beyond the traditional realm of the Red Queen, potentially explaining several important evolutionary transitions. However, these models also demonstrate that the specific consequences of coevolution are sensitive to the structure of the infection matrix, which is embedded in models to describe the likelihood of infection in encounters between specific host and parasite genotypes. Traditional cross-infection approaches to estimating infection matrices might be unreliable because evolutionary dynamics and experimental sampling lead to missing genotypes. Consequently, our goal is to identify the likely structure of infection matrices by synthesizing molecular mechanisms of host immune defense and parasite counterdefense with coevolutionary models. This synthesis reveals that the molecular mechanisms of immune reactions, although complex and diverse, conform to two basic models commonly used within coevolutionary theory: matching infection and targeted recognition. Our synthesis also overturns conventional wisdom, revealing that the general models are not taxonomically restricted but are applicable to plants, invertebrates, and vertebrates. Finally, our synthesis identifies several important areas for future research that should improve the explanatory power of coevolutionary models. The most important among these include empirical studies to identify the molecular hotspots of genotypic specificity and theoretical studies examining the consequences of matrices that more accurately represent multistep infection processes and quantitative defenses.


Non-Hodgkin's lymphoma and auricular hypoplasia: associated with juvenile colloid milium or ligneous conjunctivitis?
The exact nature of amyloid-like hyaline material deposits in the skin is not well understood in some disorders. Three of those--ligneous conjunctivitis, ligneous periodontitis and colloid milium--have been rarely reported in a same patient. We report a case of mucosal and skin deposits of an amyloid-like homogeneous material associated with non-Hodgkin's lymphoma and congenital auricular hypoplasia. We discussed and reviewed the literature on these unique associations to determine whether these are the same pathological process. We also noted whether this case represents a new syndrome or a coincidental association.


Treatment of cancer-associated venous thromboembolism in patients under palliative care.
Many patients with cancer require palliative care at some stage and the vast majority of people followed in palliative care are cancer patients. Patients with cancer are at high risk of venous thromboembolism (VTE), and this is particularly true during the advanced palliative phase when mobility is limited or absent. Patients with cancer in palliative cancer are at higher bleeding risk compared to non-cancer patients. Decisions to treat VTE or withhold anticoagulation for these patients have proven to be difficult and depend largely on an individual clinician's judgment. For this reason, we have developed a consensus proposal for appropriate management of cancer-associated thromboembolism (CAT) in patients in palliative care, which is presented in this article. The proposal was informed by the recent scientific literature retrieved through a systematic literature review. In cancer patients in advanced palliative care, the benefit-risk ratio of anticoagulation seems unfavourable with a higher haemorrhagic risk than the benefit associated with prevention of CAT recurrence and, above all, in the absence of any benefit on quality of life. For this reason, we recommend that patients should be prescribed anticoagulants on a case-by-case basis. The choice of whether to treat, and with which type of treatment, should take into account anticipated life expectancy and patient preferences, as well as clinical factors such as the estimated bleeding risk, the type of VTE experienced and the time since the VTE event.


Optical Coherence Tomography Features of Macular Hyperpigmented Lesions without Intraretinal Hyperreflective Foci in Age-Related Macular Degeneration.
To evaluate the optical coherence tomography (OCT) features of hyperpigmented lesions in the absence of intraretinal hyperreflective foci (IHRF) on OCT in eyes with age-related macular degeneration (AMD). We retrospectively analyzed OCT images of eyes with intermediate AMD (iAMD) and macular hyperpigmentation (HP) on color fundus photograph (CFP) but without IHRF on OCT in the corresponding location. The most prominent or definite HP was selected for analysis. The infrared reflectance (IR) image registered with the CFP, and the location corresponding to the HP lesion were defined on the IR image. The location of the HP on the corresponding OCT B-scan was assessed for retinal pigment epithelium (RPE) elevation, acquired vitelliform lesion (AVL), abnormal retinal pigment epithelium + basal lamina (RPE + BL) band reflectivity, RPE + BL band thickening, as well as interdigitation zone (IZ), ellipsoid zone (EZ) and external limiting membrane (ELM) disruption. 49 eyes (39 patients) were included in this study. Forty-six (94%) of the hyperpigmented lesions showed a thickened RPE + BL band. RPE + BL band reflectivity was increased in 37 (76%) of the lesions. RPE + BL band thickening, however, was not correlated with RPE + BL band reflectivity (p-value = 0.31). Either thickening or hyperreflectivity of the RPE + BL band was present in all cases. Twenty (41%) lesions had evidence of ELM disruption, 42 (86%) demonstrated EZ disruption and 48 (98%) had IZ disruption. Five (10%) HPs demonstrated AVL. Among cases with RPE elevation (15 cases, 31%), 10 were classified as drusen, 2 as drusenoid PEDs, and 3 as fibrovascular PEDs. Thickening and/or hyperreflectivity of the RPE + BL band commonly correspond to regions of macular hyperpigmentation without IHRF in eyes with iAMD.


Is anterior cingulate cortex necessary for cognitive control?
Functional neuroimaging studies in normal humans suggest that dorsal anterior cingulate cortex (dACC) plays an important role in cognitive control. This brain area is reliably activated when tasks require the ongoing adjustment of the allocation of attention. The dACC has come to occupy a central role in theories of attention and cognitive control, which hold that dACC either monitors response conflict, signalling the need for adjustments in cognitive processes, or directly mediates such adjustments. However, functional imaging results cannot establish that a brain area is necessary for a particular cognitive process. This requires evidence from loss-of-function studies. Here we assessed cognitive control in four human subjects with damage to dACC and 12 age- and education-matched control subjects using several measures drawn from the functional imaging literature. All four subjects with dACC damage showed normal adjustments in performance following manipulation of response conflict in both Stroop and go-no go tasks. Furthermore, damage to the dACC did not impair the phenomenon of post-error slowing, nor alter the ability to adjust performance in response to explicit speed or accuracy instructions. Thus, cognitive control, as assessed by four different measures in two different tasks, appears to be intact in these subjects, arguing against a necessary role for dACC in this process.


Medical treatment of critical limb ischemia: current state and future directions.
Atherosclerotic critical limb ischemia (CLI) is manifested by ischemic rest pain, non-healing ulcers or gangrene. The incidence of CLI is estimated to be approximately 500-1000 new cases per year per million people and is expected to grow in developed countries as the population ages with an increasing prevalence of diabetes. Patients diagnosed with CLI are at very high risk of major amputation and cardiovascular morbidity and mortality and experience poor physical function and quality of life. The goals of treatment for CLI are relieving ischemic pain, healing ulcers, preventing limb loss, improving patient function and quality of life, and prolonging survival. Prompt surgical or endovascular revascularization is currently recommended for limb salvage in CLI. All patients with CLI should receive cardiovascular risk reduction therapies, focused on optimizing antiplatelet therapy and risk factor management, to reduce cardiovascular event rates. Adjunctive pharmacotherapy with antithrombotic drugs, statins, and beta-blockers is critical to decrease perioperative cardiovascular complications in patients undergoing surgical vascular reconstruction and enhance postrevascularization arterial and graft patency. In non-reconstructable patients with stable pain and tissue loss, evidence suggests that prostanoids, dedicated wound care programs, and several mechanical devices, such as spinal cord stimulation, intermittent pneumatic compression, and hyperbaric oxygen therapy, can alleviate ischemic symptoms and improve limb salvage. Current medical armamentarium used in treating ischemic wounds also includes ultrasound and negative pressure wound therapy. Therapeutic neovascularization, including gene- and cell-based approaches, is a novel promising tool in the management of CLI under ongoing investigation.


The WblC/WhiB7 Transcription Factor Controls Intrinsic Resistance to Translation-Targeting Antibiotics by Altering Ribosome Composition.
Bacteria that encounter antibiotics can efficiently change their physiology to develop resistance. This intrinsic antibiotic resistance is mediated by multiple pathways, including a regulatory system(s) that activates specific genes. In some Streptomyces and Mycobacterium spp., the WblC/WhiB7 transcription factor is required for intrinsic resistance to translation-targeting antibiotics. Wide conservation of WblC/WhiB7 within Actinobacteria indicates a critical role of WblC/WhiB7 in developing resistance to such antibiotics. Here, we identified 312 WblC target genes in Streptomyces coelicolor, a model antibiotic-producing bacterium, using a combined analysis of RNA sequencing and chromatin immunoprecipitation sequencing. Interestingly, WblC controls many genes involved in translation, in addition to previously identified antibiotic resistance genes. Moreover, WblC promotes translation rate during antibiotic stress by altering the ribosome-associated protein composition. Our genome-wide analyses highlight a previously unappreciated antibiotic resistance mechanism that modifies ribosome composition and maintains the translation rate in the presence of sub-MIC levels of antibiotics.IMPORTANCE The emergence of antibiotic-resistant bacteria is one of the top threats in human health. Therefore, we need to understand how bacteria acquire resistance to antibiotics and continue growth even in the presence of antibiotics. Streptomyces coelicolor, an antibiotic-producing soil bacterium, intrinsically develops resistance to translation-targeting antibiotics. Intrinsic resistance is controlled by the WblC/WhiB7 transcription factor that is highly conserved within Actinobacteria, including Mycobacterium tuberculosis Here, identification of the WblC/WhiB7 regulon revealed that WblC/WhiB7 controls ribosome maintenance genes and promotes translation in the presence of antibiotics by altering the composition of ribosome-associated proteins. Also, the WblC-mediated ribosomal alteration is indeed required for resistance to translation-targeting antibiotics. This suggests that inactivation of the WblC/WhiB7 regulon could be a potential target to treat antibiotic-resistant mycobacteria.


The cytogenetic investigation on peripheral blood lymphocytes of a patient with thyroid cancer.
The cytogenetic investigation of a patient with follicular cancer in the left lobe of the thyroid gland was made. The study was carried out on blood lymphocytes as described earlier (1) before and after hemithyroidectomy and in different terms during the therapeutic treatment. The great number of cells with polyploid and hyperaneuploid (13% and 5% correspondingly) were revealed in the patient on the day before the operation and confirmed malignant character of the tumour, which was confirmed histologically. It is believed that cytogenetic markers as trisomies 4-th chromosome or/and X chromosome, detected in hyperaneuploids cells before the beginning of a treatment, might be specific markers for follicular thyroid cancer (FTC). The cytogenetic examination of the patient, carried out on peripheral blood lymphocytes in dynamics, allowed to observe changes of background chromosomal breaches under influence of surgical and therapeutic treatment and to estimate their efficiency.


Iguratimod prevents ovariectomy‑induced bone loss and suppresses osteoclastogenesis via inhibition of peroxisome proliferator‑activated receptor‑γ.
Iguratimod is known for its anti‑inflammatory activities and therapeutic effects in patients with rheumatoid arthritis. It has previously been demonstrated that iguratimod attenuates bone destruction and osteoclast formation in the Walker 256 rat mammary gland carcinoma cell‑induced bone cancer pain model. Therefore, it was hypothesized that iguratimod may additionally exhibit therapeutic effects on benign osteoclast‑associated diseases including postmenopausal osteoporosis. In the present study, ovariectomized mice were used to investigate the effects of iguratimod in vivo. Bone marrow mononuclear cells were cultured to detect the effects of iguratimod on receptor activator of nuclear factor‑κB ligand (RANKL)‑induced osteoclastogenesis in vitro and the molecular mechanisms involved. It was demonstrated that iguratimod may prevent ovariectomy‑induced bone loss by suppressing osteoclast activity in vivo. Consistently, iguratimod may inhibit RANKL‑induced osteoclastogenesis and bone resorption in primary bone marrow mononuclear cells. At the molecular level, peroxisome proliferator‑activated receptor‑γ (PPAR‑γ)/c‑Fos pathway, which is essential in RANKL‑induced osteoclast differentiation, was suppressed by iguratimod. Subsequently, iguratimod decreased the expression of nuclear factor of activated T cells c1 and downstream osteoclast marker genes. The results of the present study demonstrated that iguratimod may inhibit ovariectomy‑induced bone loss and osteoclastogenesis by modulating RANKL signaling. Therefore, iguratimod may act as a novel therapeutic to prevent postmenopausal osteoporosis.


Prevalence, distribution, and sequence diversity of hmwA among commensal and otitis media non-typeable Haemophilus influenzae.
Nontypeable Haemophilus influenzae (NTHi) are Gram-negative coccobacilli that colonize the human pharynx, their only known natural reservoir. Adherence to the host epithelium facilitates NTHi colonization and marks one of the first steps in NTHi pathogenesis. Epithelial cell attachment is mediated, in part, by a pair of high molecular weight (HMW) adhesins that are highly immunogenic, antigenically diverse, and display a wide range of amino acid diversity both within and between isolates. In this study, the prevalence of hmwA, which encodes the HMW adhesin, was determined for a collection of 170 NTHi isolates recovered from the middle ears of children with otitis media (OM isolates) or throats or nasopharynges of healthy children (commensal isolates) from Finland, Israel, and the U.S. Overall, hmwA was detected in 61% of NTHi isolates and was significantly more prevalent (P=0.004) among OM isolates than among commensal isolates; the prevalence ratio comparing hmwA prevalence among ear isolates with that of commensal isolates was 1.47 (95% CI (1.12, 1.92)). Ninety-five percent (98/103) of the hmwA-positive NTHi isolates possessed two hmw loci. To advance our understanding of hmwA binding sequence diversity, we determined the DNA sequence of the hmwA binding region of 33 isolates from this collection. The average amino acid identity across all hmwA sequences was 62%. Phylogenetic analyses of the hmwA binding revealed four distinct sequence clusters, and the majority of hmwA sequences (83%) belonged to one of two dominant sequence clusters. hmwA sequences did not cluster by chromosomal location, geographic region, or disease status.


Efficacy and safety of histrelin subdermal implant in children with central precocious puberty: a multicenter trial.
GnRH analog (GnRHa) therapy for central precocious puberty (CPP) typically involves im injections. The histrelin implant is a new treatment that provides a continuous slow release of the GnRHa histrelin. The objective of the study was to investigate the safety and efficacy of the subdermal histrelin implant for the treatment of CPP in treatment naive and previously treated children. This was a phase III, open-label, prospective study of 1-yr duration. The study was conducted at nine U.S. medical centers. Girls ages 2-8 yr (naive) or 2-10 yr (previously treated) and boys 2-9 yr (naive) or 2-11 yr (previously treated) with clinical evidence of CPP and a pretreatment pubertal response to leuprolide stimulation were eligible. A 50-mg histrelin implant was inserted sc in the inner upper arm. Peak LH after GnRHa stimulation testing and estradiol (girls) and testosterone (boys) were the main outcome measures. Thirty-six subjects (20 naive) were enrolled. By 1 month, peak LH fell from 28.2 +/- 19.97 (naive) to 0.8 +/- 0.39 mIU/ml (P < 0.0001) and from 2.1 +/- 2.15 (previously treated) to 0.5 +/- 0.32 mIU/ml (P < 0.0056). Estradiol suppressed from 24.5 +/- 22.27 (naive) to 5.9 +/- 2.37 pg/ml (P = 0.0016) and remained suppressed in previously treated subjects, as did testosterone. Suppression was maintained throughout the study. No significant adverse events occurred. The subdermal histrelin implant achieves and maintains excellent suppression of peak LH and sex steroid levels for 1 yr in children with CPP. The treatment is well tolerated. Long-term studies are needed to confirm these results.


Temporal motor coordination in the ankle joint following upper motor neuron lesions.
[Purpose] We compared ankle temporal motor coordination between stroke, spinal disease and healthy elderly groups, and investigated the relationship between motor impairments and gait speed. [Subjects] Twenty-four patients with stroke, 19 post-operative spinal disease patients and 17 healthy elderly subjects participated. [Methods] Ankle temporal motor coordination of the three groups was assessed using the simple reaction time, the foot-tapping test, and a rhythm task. Rhythm error and rhythm variation were analyzed using the results of the rhythm task. Isometric muscle strength, spasticity, muscle stiffness, somatosensory and 10-m gait speed of the stroke and spinal disease subjects were also measured. [Results] Only the stroke group showed significant reductions in temporal accuracy and consistency in the rhythm task. Simple reaction time and the rhythm task were significantly poorer in the stroke group, whereas the foot-tapping test was not. Stepwise multiple regression analysis indicated gait speed was explained by rhythm error and plantarflexor strength in the stroke group, and rhythm error and simple reaction time in the spinal disease group. [Conclusion] Poor performance in simple reaction time and the rhythm task in the stroke group suggest these tasks are controlled by the supraspinal central nervous system. Negative features, particularly motor coordination, are more associated with gait speed than positive features.


Survival, Persistence, and Isolation of the Emerging Multidrug-Resistant Pathogenic Yeast Candida auris on a Plastic Health Care Surface.
The emerging multidrug-resistant pathogenic yeast Candida auris represents a serious threat to global health. Unlike most other Candida species, this organism appears to be commonly transmitted within health care facilities and causes health care-associated outbreaks. To better understand the epidemiology of this emerging pathogen, we investigated the ability of C. auris to persist on plastic surfaces common in health care settings compared with that of Candida parapsilosis, a species known to colonize the skin and plastics. Specifically, we compiled comparative and quantitative data essential to understanding the vehicles of spread and the ability of both species to survive and persist on plastic surfaces under controlled conditions (25°C and 57% relative humidity), such as those found in health care settings. When a test suspension of 104 cells was applied and dried on plastic surfaces, C. auris remained viable for at least 14 days and C. parapsilosis for at least 28 days, as measured by CFU. However, survival measured by esterase activity was higher for C. auris than C. parapsilosis throughout the 28-day study. Given the notable length of time Candida species survive and persist outside their host, we developed methods to more effectively culture C. auris from patients and their environment. Using our enrichment protocol, public health laboratories and researchers can now readily isolate C. auris from complex microbial communities (such as patient skin, nasopharynx, and stool) as well as environmental biofilms, in order to better understand and prevent C. auris colonization and transmission.


Protective effect of puerarin on diabetic retinopathy in rats.
Puerarin is a major active ingredient extracted from the traditional Chinese medicine Ge-gen. The purpose of this study is to investigate the protective effect of puerarin on diabetic retinopathy (DR) and its mechanisms in rats. Seventy-two male Wistar rats were selected and divided at random into three main groups: control group, streptozotocin (STZ) group and puerarin + STZ group. Retinal histopathological observation and electron microscopic examination were performed; retinal vascular endothelial growth factor (VEGF) and hypoxia inducible factor 1 (HIF-1alpha) gene expressions were examined by Reverse transcription-polymerase chain reaction (RT-PCR) analysis. Results showed that the DR induced by STZ was significantly reduced by the treatment of puerarin as judged by the reduction of morphological changes of inner nuclear layer and outer nuclear layer at any time-point. Puerarin regulates expressions of VEGF and HIF-1alpha stimulated by STZ. It was concluded that puerarin exerts significant protective effects against DR in rats, likely regulating angiogenesis factors expressions, and thus may be an effective and promising medicine for treatment of DR.


Speculum opening in transsphenoidal surgery.
To assess the extent to which the transsphenoidal speculum can be safely opened at the face of and within the sphenoid sinus without risking damage to the optic nerves in the optic canals and at the orbital apex and the nerves coursing adjacent the walls of the sphenoid sinus. The distance was measured between the optic nerves at the level of the anterior wall of the sphenoid sinus and 0.5 and 1.0 cm within the sinus. In addition, the distance between the middle turbinates and the contralateral optic canals was assessed because this turbinate is the largest structure blocking access to the sphenoid sinus in the transsphenoidal approach and tends to force the speculum away from the midline and toward the optic nerve in the contralateral side of the approach. Opening the transsphenoidal speculum at the anterior wall of the sphenoid sinus beyond 2.5 centimeters carries some risk of damaging the optic nerves and this distance narrows when the speculum opening is positioned inside the sphenoid sinus. Displacement of the speculum to one side by the middle turbinate places the speculum near the contralateral optic nerve and may be associated with optic nerve injury with lesser degrees of speculum opening. Careful attention should be directed to avoiding excessive opening of the transsphenoidal speculum at the anterior face of the sphenoid or within the sphenoid sinus.