Boosting the Immune System for HIV Cure: A γδ T Cell Perspective.
The major barrier to HIV cure is a population of long-lived cells that harbor latent but replication-competent virus, are not eliminated by antiretroviral therapy (ART), and remain indistinguishable from uninfected cells. However, ART does not cure HIV infection, side effects to treatment still occur, and the steady global rate of new infections makes finding a sustained ART-free HIV remission or cure for HIV-seropositive individuals urgently needed. Approaches aimed to cure HIV are mostly based on the "shock and kill" method that entails the use of a drug compound to reactivate latent virus paired together with strategies to boost or supplement the existing immune system to clear reactivated latently infected cells. Traditionally, these strategies have utilized CD8+ cytotoxic lymphocytes (CTL) but have been met with a number of challenges. Enhancing innate immune cell populations, such as γδ T cells, may provide an alternative route to HIV cure. γδ T cells possess anti-viral and cytotoxic capabilities that have been shown to directly inhibit HIV infection and specifically eliminate reactivated, latently infected cells in vitro. Most notably, their access to immune privileged anatomical sites and MHC-independent antigen recognition may circumvent many of the challenges facing CTL-based strategies. In this review, we discuss the role of γδ T cells in normal immunity and HIV infection as well as their current use in strategies to treat cancer. We present this information as means to speculate about the utilization of γδ T cells for HIV cure strategies and highlight some of the fundamental gaps in knowledge that require investigation.


Structure of nanoscale-pitch helical phases: blue phase and twist-bend nematic phase resolved by resonant soft X-ray scattering.
Periodic structures of phases with orientational order of molecules but homogenous electron density distribution: a short pitch cholesteric phase, blue phase and twist-bend nematic phase, were probed by resonant soft X-ray scattering (RSoXS) at the carbon K-edge. The theoretical model shows that in the case of a simple heliconical nematic structure, two resonant signals corresponding to the full and half pitch band should be present, while only the full pitch band is observed experimentally. This suggests that the twist-bend nematic phase has a complex structure with a double-helix built of two interlocked, shifted helices. We confirm that the helical pitch in the twist-bend nematic phase is in a 10 nm range for both the chiral and achiral materials. We also show that the symmetry of the blue phase can be unambiguously determined through a resonant enhancement of the X-ray diffraction signals, by including polarization effects, which are found to be an important indicator in phase structure determination.


N-acetylcysteine: multiple clinical applications.
N-acetylcysteine is the acetylated variant of the amino acid L-cysteine and is widely used as the specific antidote for acetaminophen overdose. Other applications for N-acetylcysteine supplementation supported by scientific evidence include prevention of chronic obstructive pulmonary disease exacerbation, prevention of contrast-induced kidney damage during imaging procedures, attenuation of illness from the influenza virus when started before infection, treatment of pulmonary fibrosis, and treatment of infertility in patients with clomiphene-resistant polycystic ovary syndrome. Preliminary studies suggest that N-acetylcysteine may also have a role as a cancer chemopreventive, an adjunct in the eradication of Helicobacter pylori, and prophylaxis of gentamicin-induced hearing loss in patients on renal dialysis.


The oligomeric state of the truncated mechanosensitive channel of large conductance shows no variance in vivo.
The mechanosensitive channel of large conductance (MscL) from E. coli serves as an emergency release valve allowing the cell to survive acute osmotic downshock. It is one of the best studied mechanosensitive channels and serves as a paradigm for how a protein can sense and respond to membrane tension. Two MscL crystal structures of the orthologs M. tuberculosis and S. aureus have been solved showing pentameric and tetrameric structures, respectively. Several studies followed to understand whether the discrepancy in their stoichiometry was a species difference or a consequence of the protein manipulation for crystallization. Two independent studies now agree that the full-length S. aureus MscL is actually a pentamer, not tetramer. While detergents appear to play a role in modifying the oligomeric state of the protein, a cytoplasmic helical bundle has also been implicated. Here, we evaluate the role of the C-terminal region of S. aureus MscL in the oligomerization of the channel in native membranes by using an in vivo disulfide-trapping technique. We find that the oligomeric state of S. aureus MscLs with different C-terminal truncations, including the one used to obtain the tetrameric S. aureus MscL crystal structure, are pentamers in vivo. Thus, the C-terminal domain of the S. aureus protein only plays a critical role in the oligomeric state of the SaMscL protein when it is solubilized in detergent.


Septic Arthritis of the Shoulder with Proximal Humerus Osteomyelitis, Treated by Ilizarov Shoulder Arthrodesis.
Septic arthritis of the shoulder can destroy the glenohumeral joint resulting in significant pain, instability and poor function. Surgical treatment is notoriously difficult with significant risk of complications. Shoulder arthrodesis has been performed using both internal and external fixation but has high complication rates in the presence of infection. We present our experience of managing a patient with significant pain and loss of function as a result of septic arthritis of the shoulder following infected proximal humeral fracture fixation with associated proximal humeral osteomyelitis. We discuss patient presentation, appropriate investigations, decision-making, pre-operative planning and the principles of managing of bone and joint infection. Wide excision with Ilizarov stabilisation and implantation of an absorbable antibiotic carrier, allowed successful fusion with eradication of the infection. The benefits of using this arthrodesis technique include reduced risk of infection recurrence and excellent stability of construct, therefore reducing time to fusion and minimising soft tissue trauma with the opportunity for early rehabilitation and return to optimal level of function.


CNS-specific ablation of steroidogenic factor 1 results in impaired female reproductive function.
The ventromedial hypothalamic nucleus (VMH) regulates a variety of homeostatic processes including female sexual behavior and reproduction. In the current study, we assessed the roles of steroidogenic factor 1 (SF-1) on reproductive function in the VMH using central nervous system-specific SF-1 knockout (SF-1 KO(nCre;F/-)) mice. Here we show that SF-1 KO(nCre;F/-) females exhibited marked impairment in female reproduction. Although male mice appeared to be normal in all aspects studied, including sexual behavior, SF-1 KO(nCre;F/-) females showed infertility or subfertility. Although adult SF-1 KO(nCre;F/-) females showed decreased or lacked corpora lutea, exogenous administration of gonadotropins induced the formation of multiple corpora lutea and induced normal ovulation, demonstrating that the ovaries are functionally intact. In addition, SF-1 KO(nCre;F/-) females stimulated with a synthetic GnRH agonist after priming exhibited markedly reduced LH secretion compared with wild-type littermates, arguing that disorganization in and around the VMH caused by SF-1 ablation interferes with the GnRH priming process or gonadotrope LH capacity. Furthermore, the SF-1 KO(nCre;F/-) females primed with estrogen benzoate and progesterone failed to induce steroid receptors around the VMH, consistent with impaired lordosis behavior in the SF-1 KO(nCre;F/-) females. Collectively, our results highlight that SF-1 in the VMH plays crucial roles in regulation of female reproductive function, presumably by organizing a precise neuronal connection and communication in and around the VMH.


Geographic Clustering of Admissions to Inpatient Psychiatry among Adults with Cognitive Disorders in Ontario, Canada: Does Distance to Hospital Matter?
This study examined relationships among hospital accessibility, socio-economic context, and geographic clustering of inpatient psychiatry admissions for adults with cognitive disorders in Ontario, Canada. A retrospective cross-sectional analysis was conducted using admissions data from 71 hospitals with inpatient psychiatry beds in Ontario, Canada between 2011 and 2014. Data included 7,637 unique admissions for 4,550 adults with a DSM-IV diagnosis of Delirium, Dementia, Amnestic and other Cognitive Disorders. Bayesian spatial Poisson regression was employed to examine the relationship between accessibility of general hospitals with psychiatric beds and psychiatric hospitals, area-level marginalization, and hospitalization rate with the risk of admission to inpatient psychiatry among adults with cognitive disorders across 516 Forward Sortation Areas (FSA) in Ontario. Residential instability and the overall hospitalization rate were significantly associated with an increase in the relative risk of admissions to inpatient psychiatry. Accessibility to general hospitals and psychiatric hospitals were marginally insignificant at the 95% credible interval in the final model. Significant geographic clustering of admissions was identified where individuals residing in FSA's with the highest relative risk were 2.0 to 7.1 times more likely to be admitted to inpatient psychiatry compared to the average. Geographic clustering of inpatient psychiatry admissions for adults with cognitive disorders exists across the Province of Ontario, Canada. At the geographic level, the risk of admission was positively associated with residential instability and the overall hospitalization rate, but not distance to the closest general or psychiatric hospital.


High-density genetic linkage map construction and identification of fruit-related QTLs in pear using SNP and SSR markers.
Pear (Pyrus spp) is an important fruit crop, grown in all temperate regions of the world, with global production ranked after grape and apples among deciduous tree crops. A high-density linkage map is a valuable tool for fine mapping quantitative trait loci (QTL) and map-based gene cloning. In this study, we firstly constructed a high-density linkage map of pear using SNPs integrated with SSRs, developed by the rapid and robust technology of restriction-associated DNA sequencing (RADseq). The linkage map consists of 3143 SNP markers and 98 SSRs, 3241 markers in total, spanning 2243.4 cM, with an average marker distance of 0.70 cM. Anchoring SSRs were able to anchor seventeen linkage groups to their corresponding chromosomes. Based on this high-density integrated pear linkage map and two years of fruit phenotyping, a total of 32 potential QTLs for 11 traits, including length of pedicel (LFP), single fruit weight (SFW), soluble solid content (SSC), transverse diameter (TD), vertical diameter (VD), calyx status (CS), flesh colour (FC), juice content (JC), number of seeds (NS), skin colour (SC), and skin smooth (SS), were identified and positioned on the genetic map. Among them, some important fruit-related traits have for the first time been identified, such as calyx status, length of pedicel, and flesh colour, and reliable localization of QTLs were verified repeatable. This high-density linkage map of pear is a worthy reference for mapping important fruit traits, QTL identification, and comparison and combination of different genetic maps.


The nanomechanics of individual proteins.
Mechanical forces regulate a large variety of cellular functionalities, encompassing e.g. motility, differentiation and muscle contractility. To adapt to the dynamic change in mechanical stress, the constitutive individual proteins need to reversibly stretch and recoil over long periods of time. Yet, the molecular mechanisms controlling the mechanical unfolding and refolding of proteins cannot be accessed by protein folding biochemistry experiments conducted in the bulk, because they cannot typically apply forces to individual proteins. The advent of single-molecule nanomechanical techniques, often combined with bespoke protein engineering strategies, has enabled monitoring the conformational dynamics of proteins under force with unprecedented length-, time- and force-resolution. This review focuses on the fundamental operational principles of the main single-molecule nanomechanical techniques, placing particular emphasis on the most common analytical approaches used to extract information directly from the experiments. The breadth of enabling applications highlights the most exciting and promising outputs from the nanomechanics field to date.


Motives and Barriers to Physical Activity Among Older Adults of Different Socioeconomic Status.
This study explored motives and barriers to physical activity (PA) among older adults of differing socioeconomic status (SES) utilizing a self-determination theory and self-efficacy theory framework. Focus groups (n = 4) were conducted with older adults (n = 28) from two SES groups, using thematic analysis to identify motives and barriers. Integrated and identified regulations and, to a lesser extent, intrinsic motives, were evident across SES groups. Verbal persuasion and affective and physiological states emerged as prominent efficacy sources regardless of SES. More barriers were reported by the low SES group, with health conditions, neighborhood safety, and PA guidelines knowledge emerging as most salient. Time emerged as a prominent barrier for the high SES group. Integrated and identified regulations should be fostered in future interventions and policy regardless of SES. Barriers to PA varied across SES groups; thus future interventions and policy should account for such differences.


Antenatal exposure to chorioamnionitis affects lipid metabolism in 7-week-old sheep.
Antenatal exposure of the fetus to inflammation may alter postnatal organ development. In our previous work, we demonstrated that the fetal liver is involved in the systemic inflammation associated with chorioamnionitis, leading to metabolic changes. On the basis of these findings, we hypothesized that chorioamnionitis can lead to postnatal inflammation-related liver injury and disturbed lipid metabolism. Chorioamnionitis was induced in sheep by intra-amniotic injection of lipopolysaccharide (LPS) or saline at 90, 100 and 110 days of gestation. Liver homeostasis and lipid metabolism were analyzed at term and at 7 weeks of age. At term, hepatic T-lymphocytes and apoptotic hepatocytes were increased. In addition, hepatic cholesterol and triglyceride levels were decreased in LPS-exposed animals compared with controls. At 7 weeks of age, no hepatic inflammation could be detected. However, liver triglycerides and plasma cholesterol levels were increased in LPS-exposed animals relative to controls. The changes in lipid levels at 7 weeks of age were associated with increased leptin receptor mRNA levels, increased lipid peroxidation, increased expression of cytochrome c oxidase subunit 4 as a marker for mitochondrial function and increased circulating ceramide levels. These findings demonstrate that chorioamnionitis-mediated antenatal inflammation-related liver disturbances have long-lasting postnatal effects on lipid metabolism.


Menarche and the onset of depression and anxiety in Victoria, Australia.
Psychiatric disorder often begins at adolescence. This study aimed to examine the associations between puberty and social circumstances and the adolescent rise in depression and anxiety. A two stage cluster sampling procedure was used to identify a representative group of Australian secondary school students in years 7 (age 12-13 years), 9 (14-15 years), and 11 (16-17 years) of 45 Victorian schools. The computerised clinical interview schedule (CIS) was used to evaluate psychiatric morbidity. A total of 2525 subjects completed the survey - an overall participation rate of 83%. Levels of depression and anxiety increased with the secondary school years and girls had significantly higher rates at each school year level. For boys, the clearest independent associations with depression and anxiety were rising school year level and high parental educational achievement. For girls menarchal status emerged as the strongest predictor. Associations with age and school year level, evident on univariate analysis, did not persist when the recency of menarche was taken into account. After addition of measures of perceived social stress to a multivariate model, a significant association between depression/anxiety and parental divorce disappeared but the association with menarche persisted. Menarche marks a transition in the risk of depression and anxiety in girls. The pattern of findings is consistent with a biological mediation of this association.


Recombinant human interleukin 4 has antiproliferative activity on human tumor cell lines derived from epithelial and nonepithelial histologies.
Interleukin 4, a T cell-derived 20-kDa glycoprotein, plays an important role in regulating the immune response of B cells, T cells, and macrophages against infections and malignant cells. For this reason recombinant human interleukin 4 (rhIL-4) has entered early clinical trials in cancer patients. In the present study we report that rhIL-4 has an antiproliferative effect on five of nine cell lines derived from human colon tumors, head and neck tumors, and glioblastomas as measured by a decrease of colony formation in human tumor cloning assays. All of the cell lines with in vitro responsiveness express at least 100 high-affinity receptors for human interleukin 4 per cell on their cell surface, whereas the nonresponsive tumor cell lines lack expression of high-affinity receptors for human interleukin 4 on their cell surface. In the next series of experiments we have xenotransplanted some of the responsive cell lines into athymic nude mice. Subsequently, the animals were treated s.c. twice daily with 0.5 mg/m2 rhIL-4 or control vehicle for at least 12 days. There was a clear growth inhibition of these xenotransplanted tumors in the mice treated with rhIL-4. Histology of the tumors in both groups revealed no marked infiltration with murine hematopoietic and lymphocytic cells as evaluated by staining with a rat anti-mouse CD45 antibody. We conclude that rhIL-4 has a direct therapeutic activity on the growth of some human epithelial and nonepithelial tumor cell lines which, along with its regulatory function on hematolymphopoietic cells, makes this cytokine an interesting candidate for experimental tumor therapy.


Engineering of a novel Saccharomyces cerevisiae wine strain with a respiratory phenotype at high external glucose concentrations.
The recently described respiratory strain Saccharomyces cerevisiae KOY.TM6*P is, to our knowledge, the only reported strain of S. cerevisiae which completely redirects the flux of glucose from ethanol fermentation to respiration, even at high external glucose concentrations (27). In the KOY.TM6*P strain, portions of the genes encoding the predominant hexose transporter proteins, Hxt1 and Hxt7, were fused within the regions encoding transmembrane (TM) domain 6. The resulting chimeric gene, TM6*, encoded a chimera composed of the amino-terminal half of Hxt1 and the carboxy-terminal half of Hxt7. It was subsequently integrated into the genome of an hxt null strain. In this study, we have demonstrated the transferability of this respiratory phenotype to the V5 hxt1-7Delta strain, a derivative of a strain used in enology. We also show by using this mutant that it is not necessary to transform a complete hxt null strain with the TM6* construct to obtain a non-ethanol-producing phenotype. The resulting V5.TM6*P strain, obtained by transformation of the V5 hxt1-7Delta strain with the TM6* chimeric gene, produced only minor amounts of ethanol when cultured on external glucose concentrations as high as 5%. Despite the fact that glucose flux was reduced to 30% in the V5.TM6*P strain compared with that of its parental strain, the V5.TM6*P strain produced biomass at a specific rate as high as 85% that of the V5 wild-type strain. Even more relevant for the potential use of such a strain for the production of heterologous proteins and also of low-alcohol beverages is the observation that the biomass yield increased 50% with the mutant compared to its parental strain.


Immunization of rabbits with recombinant Treponema pallidum surface antigen 4D alters the course of experimental syphilis.
Rabbits immunized with the protease resistant recombinant Treponema pallidum surface-associated Ag 4D showed an altered course of experimental syphilis after intradermal challenge with virulent T. pallidum. Vaccination trials using three different protocols were examined. In one experiment, a combined i.m.-i.v. immunization protocol was compared with an i.m. injection schedule. Four of five rabbits immunized by the i.m.-i.v. protocol developed morphologically atypical lesions at each of 16 sites, whereas the i.m. injected animals showed no evidence of attenuated disease. Moreover, immunization by both protocols elicited equally high ELISA titers of 4D-specific IgG antibody suggesting that cellular immune mechanisms may have been involved. In an effort to augment cell-mediated responses to 4D, seven rabbits received i.v. immunizations with an emulsion of 4D, purified BCG cell wall skeletons, and trehalose dimycolate in an oil-microdroplet form (4D-cell wall skeleton-trehalose dimycolate). Splenic lymphocytes from three representative immunized animals had a strong in vitro proliferative response to 4D as compared to controls. Further, all animals developed high anti-4D titers in response to immunization. After challenge with virulent T. pallidum, six of the seven rabbits developed an early cutaneous response (3-8 days) at the sites of inoculation consistent with a hypersensitivity reaction followed by morphologically atypical lesions. Aspirates from each of three representative atypical lesions were devoid of treponemes by darkfield examination at day 14, whereas motile T. pallidum were observed in aspirates from four of five control sites. Between days 20 and 24, lesions in the 4D-cell wall skeleton-trehalose dimycolate immunized rabbits began to enlarge and become more typical in appearance; aspirates from six of seven representative lesions were darkfield positive. We conclude that 1) immunization with the r4D Ag alters the course of experimental syphilis in a manner consistent with previously defined parameters of partial protection and 2) immunizations protocols containing an i.v. component are more effective than protocols using the i.m. route alone.


Postshunt lateral ventricular volume, white matter integrity, and intellectual outcomes in spina bifida and hydrocephalus.
No previous reports exist that have evaluated the relationships of white matter (WM) integrity with the number of shunt revisions, ventricular volume after shunting, and cognition in medically stable children who have spina bifida and hydrocephalus (SBH). The authors hypothesized that enlarged ventricles and a greater number of shunt revisions decrease WM integrity in children. In total, 80 children (mean age 13.7 years) who had SBH underwent MRI and IQ testing. Probabilistic diffusion tractography was performed to determine mean diffusion tensor imaging (DTI) metrics along the frontal and parietal tectocortical pathways. The DTI metrics were evaluated for significant correlation with a composite IQ measure and with the total number of shunt revisions and the total lateral ventricular volume obtained through semiautomated parcellation of T1-weighted MRI scans. An enlargement in total lateral ventricle volume and an increase in the number of shunt revisions were both associated with higher fractional anisotropy (FA) and with lower radial diffusivity (RD) along both frontal and parietal tectocortical pathways. Children who had not undergone a shunt revision had on average a greater lateral ventricle volume and higher FA and lower RD along frontal and parietal pathways than those who had undergone multiple shunt revisions. The mean DTI metrics along parietal pathways predicted IQ scores, but intellectual ability was not significantly correlated with ventricular volume or with the number of lifetime shunt revisions. Significant changes in DTI metrics were observed as a function of ventricular volume. An increased lateral ventricle volume was associated with elevated FA and decreased RD. Given that the participants were medically stable at the time of the MRI examination, the results suggested that those who have enlarged ventricles show a DTI pattern consistent with axonal compression due to increased intracranial pressure (ICP) in attenuated hydrocephalus. Although limited by a cross-sectional design, the study's findings suggest that DTI metrics may serve as sensitive indicators for chronic, mild hydrocephalus in the absence of overt clinical symptoms due to increased ICP. Having enlarged ventricles and undergoing multiple shunt revisions did not affect intellectual ability in children with SBH.


Treatment of Breast Cancer With Gonadotropin-Releasing Hormone Analogs.
Although significant progress has been made in the implementation of new breast cancer treatments over the last three decades, this neoplasm annually continues to show high worldwide rates of morbidity and mortality. In consequence, the search for novel therapies with greater effectiveness and specificity has not come to a stop. Among the alternative therapeutic targets, the human gonadotropin-releasing hormone type I and type II (hGnRH-I and hGnRH-II, respectively) and its receptor, the human gonadotropin-releasing hormone receptor type I (hGnRHR-I), have shown to be powerful therapeutic targets to decrease the adverse effects of this disease. In the present review, we describe how the administration of GnRH analogs is able to reduce circulating concentrations of estrogen in premenopausal women through their action on the hypothalamus-pituitary-ovarian axis, consequently reducing the growth of breast tumors and disease recurrence. Also, it has been mentioned that, regardless of the suppression of synthesis and secretion of ovarian steroids, GnRH agonists exert direct anticancer action, such as the reduction of tumor growth and cell invasion. In addition, we discuss the effects on breast cancer of the hGnRH-I and hGnRH-II agonist and antagonist, non-peptide GnRH antagonists, and cytotoxic analogs of GnRH and their implication as novel adjuvant therapies as antitumor agents for reducing the adverse effects of breast cancer. In conclusion, we suggest that the hGnRH/hGnRHR system is a promising target for pharmaceutical development in the treatment of breast cancer, especially for the treatment of advanced states of this disease.


Alveolar nitric oxide and its role in pediatric asthma control assessment.
Nitric oxide can be measured at multiple flow rates to determine proximal (maximum airway nitric oxide flux; JawNO) and distal inflammation (alveolar nitric oxide concentration; CANO). The main aim was to study the association among symptoms, lung function, proximal (maximum airway nitric oxide flux) and distal (alveolar nitric oxide concentration) airway inflammation in asthmatic children treated and not treated with inhaled glucocorticoids. A cross-sectional study with prospective data collection was carried out in a consecutive sample of girls and boys aged between 6 and 16 years with a medical diagnosis of asthma. Maximum airway nitric oxide flux and alveolar nitric oxide concentration were calculated according to the two-compartment model. In asthmatic patients, the asthma control questionnaire (CAN) was completed and forced spirometry was performed. In controls, differences between the sexes in alveolar nitric oxide concentration and maximum airway nitric oxide flux and their correlation with height were studied. The correlation among the fraction of exhaled NO at 50 ml/s (FENO50), CANO, JawNO, forced expiratory volume in 1 second (FEV1) and the CAN questionnaire was measured and the degree of agreement regarding asthma control assessment was studied using Cohen's kappa. We studied 162 children; 49 healthy (group 1), 23 asthmatic participants without treatment (group 2) and 80 asthmatic patients treated with inhaled corticosteroids (group 3). CANO (ppb) was 2.2 (0.1-4.5), 3 (0.2-9.2) and 2.45 (0.1-24), respectively. JawNO (pl/s) was 516 (98.3-1470), 2356.67 (120-6110) and 1426 (156-11805), respectively. There was a strong association (r=0.97) between FENO50 and JawNO and the degree of agreement was very good in group 2 and was good in group 3. There was no agreement or only slight agreement between the measures used to monitor asthma control (FEV1, CAN questionnaire, CANO and JawNO). The results for CANO and JawNO in controls were similar to those found in other reports. There was no agreement or only slight agreement among the three measure instruments analyzed to assess asthma control. In our sample, no additional information was provided by CANO and JawNO.


Music and learning-induced cortical plasticity.
Auditory stimuli are encoded by frequency-tuned neurons in the auditory cortex. There are a number of tonotopic maps, indicating that there are multiple representations, as in a mosaic. However, the cortical organization is not fixed due to the brain's capacity to adapt to current requirements of the environment. Several experiments on cerebral cortical organization in musicians demonstrate an astonishing plasticity. We used the MEG technique in a number of studies to investigate the changes that occur in the human auditory cortex when a skill is acquired, such as when learning to play a musical instrument. We found enlarged cortical representation of tones of the musical scale as compared to pure tones in skilled musicians. Enlargement was correlated with the age at which musicians began to practice. We also investigated cortical representations for notes of different timbre (violin and trumpet) and found that they are enhanced in violinists and trumpeters, preferentially for the timbre of the instrument on which the musician was trained. In recent studies we extended these findings in three ways. First, we show that we can use MEG to measure the effects of relatively short-term laboratory training involving learning to perceive virtual instead of spectral pitch and that the switch to perceiving virtual pitch is manifested in the gamma band frequency. Second, we show that there is cross-modal plasticity in that when the lips of trumpet players are stimulated (trumpet players assess their auditory performance by monitoring the position and pressure of their lips touching the mouthpiece of their instrument) at the same time as a trumpet tone, activation in the somatosensory cortex is increased more than it is during the sum of the separate lip and trumpet tone stimulation. Third, we show that musicians' automatic encoding and discrimination of pitch contour and interval information in melodies are specifically enhanced compared to those in nonmusicians in that musicians show larger functional mismatch negativity (MMNm) responses to occasional changes in melodic contour or interval, but that the two groups show similar MMNm responses to changes in the frequency of a pure tone.


pp-GalNAc-T13 induces high metastatic potential of murine Lewis lung cancer by generating trimeric Tn antigen.
In order to analyze the mechanisms for cancer metastasis, high metastatic sublines (H7-A, H7-Lu, H7-O, C4-sc, and C4-ly) were obtained by repeated injection of mouse Lewis lung cancer sublines H7 and C4 into C57BL/6 mice. These sublines exhibited increased proliferation and invasion activity in vitro. Ganglioside profiles exhibited lower expression of GM1 in high metastatic sublines than the parent lines. Then, we established GM1-Si-1 and GM1-Si-2 by stable silencing of GM1 synthase in H7 cells. These GM1-knockdown clones exhibited increased proliferation and invasion. Then, we explored genes that markedly altered in the expression levels by DNA microarray in the combination of C4 vs. C4-ly or H7 vs. H7 (GM1-Si). Consequently, pp-GalNAc-T13 gene was identified as up-regulated genes in the high metastatic sublines. Stable transfection of pp-GalNAc-T13 cDNA into C4 (T13-TF) resulted in increased invasion and motility. Then, immunoblotting and flow cytometry using various antibodies and lectins were performed. Only anti-trimeric Tn antibody (mAb MLS128), showed increased expression levels of trimeric Tn antigen in T13-TF clones. Moreover, immunoprecipitation/immunoblotting was performed by mAb MLS128, leading to the identification of an 80 kDa band carrying trimeric Tn antigen, i.e. Syndecan-1. Stable silencing of endogenous pp-GalNAc-T13 in C4-sc (T13-KD) revealed that primary tumors generated by subcutaneous injection of T13-KD clones showed lower coalescence to fascia and peritoneum, and significantly reduced lung metastasis than control clones. These data suggested that high expression of pp-GalNAc-T13 gene generated trimeric Tn antigen on Syndecan-1, leading to the enhanced metastasis.


Expectation and attention in hierarchical auditory prediction.
Hierarchical predictive coding suggests that attention in humans emerges from increased precision in probabilistic inference, whereas expectation biases attention in favor of contextually anticipated stimuli. We test these notions within auditory perception by independently manipulating top-down expectation and attentional precision alongside bottom-up stimulus predictability. Our findings support an integrative interpretation of commonly observed electrophysiological signatures of neurodynamics, namely mismatch negativity (MMN), P300, and contingent negative variation (CNV), as manifestations along successive levels of predictive complexity. Early first-level processing indexed by the MMN was sensitive to stimulus predictability: here, attentional precision enhanced early responses, but explicit top-down expectation diminished it. This pattern was in contrast to later, second-level processing indexed by the P300: although sensitive to the degree of predictability, responses at this level were contingent on attentional engagement and in fact sharpened by top-down expectation. At the highest level, the drift of the CNV was a fine-grained marker of top-down expectation itself. Source reconstruction of high-density EEG, supported by intracranial recordings, implicated temporal and frontal regions differentially active at early and late levels. The cortical generators of the CNV suggested that it might be involved in facilitating the consolidation of context-salient stimuli into conscious perception. These results provide convergent empirical support to promising recent accounts of attention and expectation in predictive coding.


Application of 1H NMR spectroscopy to the metabolic phenotyping of rodent brain extracts: A metabonomic study of gut microbial influence on host brain metabolism.
1H NMR Spectroscopy has been applied to determine the neurochemical profiles of brain extracts from the frontal cortex and hippocampal regions of germ free and normal mice and rats. The results revealed a number of differences between germ free (GF) and conventional (CV) rats or specific pathogen-free (SPF) mice with microbiome-associated metabolic variation found to be both species- and region-dependent. In the mouse, the GF frontal cortex contained lower amounts of creatine, N-acetyl-aspartate (NAA), glycerophosphocholine and lactate, but greater amounts of choline compared to that of specific pathogen free (SPF) mice. In the hippocampus, the GF mice had greater creatine, NAA, lactate and taurine content compared to those of the SPF animals, but lower relative quantities of succinate and an unidentified lipid-related component. The GF rat frontal cortex contained higher relative quantities of lactate, creatine and NAA compared to the CV animals whilst the GF hippocampus was characterized by higher taurine and phosphocholine concentrations and lower quantities of NAA, N-acetylaspartylglutamate and choline compared to the CV animals. Of note is that, in both rat and mouse brain extracts, concentrations of hippocampal taurine were found to be greater in the absence of an established microbiome. The results provide further evidence that brain biochemistry can be influenced by gut microbial status, specifically metabolites involved in energy metabolism demonstrating biochemical dialogue between the microbiome and brain.


A mutation in the largest (catalytic) subunit of RNA polymerase II and its relation to the arrest of the cell cycle in G(1) phase.
Transcriptional activity of RNA polymerase II is modulated during the cell cycle. We previously identified a temperature-sensitive mutation in the largest (catalytic) subunit of RNA polymerase II (RPB1) that causes cell cycle arrest and genome instability. We now characterize a different cell line that has a temperature-sensitive defect in cell cycle progression, and find that it also has a mutation in RPB1. The temperature-sensitive mutant, tsAF8, of the Syrian hamster cell line, BHK21, arrests at the non-permissive temperature in the mid-G(1) phase. We show that RPB1 in tsAF8--which is found exclusively in the nucleus at the permissive temperature--is also found in the cytoplasm at the non-permissive temperature. Comparison of the DNA sequences of the RPB1 gene in the wild-type and mutant shows the mutant phenotype results from a (hemizygous) C-to-A variation at nucleotide 944 in one RPB1 allele; this gives rise to an ala-to-asp substitution at residue 315 in the protein. Aligning the amino acid sequences from various species reveals that ala(315) is highly conserved in eukaryotes.


UK Food Standards Agency cis-monounsaturated fatty acid workshop report.
The UK Food Standards Agency convened a group of expert scientists to review current research investigating the optimal dietary intake for n-9 cis-monounsaturated fatty acids (MUFA). The aim was to review the mechanisms underlying the reported beneficial effects of MUFA on CHD risk, and to establish priorities for future research. The issue of optimal MUFA intake is contingent upon optimal total fat intake; however, there is no consensus of opinion on what the optimal total fat intake should be. Thus, it was recommended that a large multi-centre study should look at the effects on CHD risk of MUFA replacement of saturated fatty acids in relation to varying total fat intakes; this study should be of sufficient size to take account of genetic variation, sex, physical activity and stage of life factors, as well as being of sufficient duration to account for adaptation to diets. Recommendations for studies investigating the mechanistic effects of MUFA were also made. Methods of manipulating the food chain to increase MUFA at the expense of saturated fatty acids were also discussed.


Chronic consumption of trans-fat-rich diet increases hepatic cholesterol levels and impairs muscle insulin sensitivity without leading to hepatic steatosis and hypertriglyceridemia in female Fischer rats.
The impact of industrial trans fatty acids (TFAs) on lipid metabolism and health remains elusive. We compared the effect of long-term (52 weeks) ingestion of 10% partially hydrogenated vegetable oil, providing 4.2% of total energy from TFAs, on hepatic lipid metabolism and muscle insulin sensitivity in weanling female Fischer rats with that of palmolein (monounsaturated fatty acid, MUFA), sunflower (n-6 polyunsaturated fatty acid, PUFA), and a blend of sunflower and fish oil (n-3 PUFA). The proportion of plasma high-density lipoprotein cholesterol in total cholesterol and reverse cholesterol transport-associated protein expressions were similar in all the groups. Despite higher lipogenic-pathway protein levels, steatosis or hypertriglyceridemia was not observed in TFA-fed rats. Though TFA ingestion had no effect on fasting plasma glucose, insulin levels or oral glucose tolerance, it significantly decreased muscle insulin-stimulated glucose uptake as compared to PUFAs. Further, TFA ingestion increased adipose tissue retinol-binding protein 4 mRNA as compared to PUFAs (n-6 or n-3). The effects of MUFA (oleic acid) on all these parameters were comparable to those observed for TFAs. Compared to PUFA-rich diets, chronic consumption of a TFA-rich diet did not lead to steatosis or hypertriglyceridemia; however, it significantly impaired muscle insulin sensitivity, while no changes were found in the oral glucose tolerance test.


Tool Life Performance of Injection Mould Tooling Fabricated by Selective Laser Melting for High-Volume Production.
Rapid Tooling processes are developing and proving to be a reliable method to compete with subtractive techniques for tool making. This paper investigates large volume production of components produced from Selective Laser Melting (SLM) fabricated injection moulding tool inserts. To date, other researchers have focused primarily on investigating the use of additive manufacturing technology for injection moulding for low-volume component production rather than high volume production. In this study, SLM technology has been used to fabricate four Stainless Steel 316L tool inserts of a similar geometry for an after-market automotive spare part. The SLM tool inserts have been evaluated to analyse the maximum number of successful injections and quality of performance. Microstructure inspection and chemical composition analysis have been investigated. Performance tests were conducted for the four tool inserts before and after injection moulding in the context of hardness testing and dimensional accuracy. For the first reported time, 150,000 injected products were successfully produced from the four SLM tool inserts. Tool inserts performance was monitored under actual operating conditions considering high-level demands. In the scope of this research, SLM proved to be a dependable manufacturing technique for most part geometries and an effective alternative to subtractive manufacturing for high-volume injection moulding tools for the aftermarket automotive sector.


Putting the systems back into systems biology.
In recent years the term "systems biology" has become widespread in the biological literature, but most of the papers in which these words appear have surprisingly little to do with older notions of biological systems: they often seem to imply little more than reductionist biology applied on a large scale, with a little attention to interactions between some of the components, but with minimal attention to the kinetic properties of enzymes, which supplied much of the reductionist foundation of biochemistry. A systemic approach to biology ought to put the emphasis on the entire system; insofar as it is concerned with components at all, it is to explain their roles in meeting the needs of the system as a whole. Genuinely systemic thinking allows us to understand how biochemical systems are regulated, and why clumsy attempts to manipulate them for biotechnological purposes may fail. At a more abstract level, it is necessary for understanding the nature of life, because as long as an organism is treated as no more than a collection of components, one cannot ask the right questions, and certainly cannot answer them.


Effects of size-controlled TiO2 nanopowders synthesized by chemical vapor condensation process on conversion efficiency of dye-sensitized solar cells.
To investigate the microstructural effects of the synthesized TiO2 nanopowders such as particle size, specific surface area, pore size and pore distributions for the application of an anode material of dye-sensitized solar cells (DSSC), size-controlled and well-dispersed TiO2 nanopowders were synthesized by chemical vapor condensation (CVC) process in the range of 800-1000 degreesC under a pressure of 50 mbar. The average particle size of synthesized TiO2 nanopowders was increased with increasing temperature from 13 nm for 800 degreesC, 15 nm for 900 degreesC and 26 nm. The specific surface area of synthesized nanoparticles were measured as 119.1 m2/g for 800 degreesC, 104.7 m2/g for 900 degreesC and 59.5 m2/g for 1000 degreesC, respectively. The conversion efficiency values (eta%) of DSSC with the synthesized TiO2 nanopowders at 800 degreesC, 900 degreesC, and 1000 degreesC were 2.59%, 5.96% and 3.66%, respectively. The highest conversion efficiency obtained in the 900 degreesC (5.96%) sample is thought to be attributable to homogeneous particle size and pore distributions, large specific surface area, and high transmittance in regions of dye absorption wavelength.


Video Rating in Neurodegenerative Disease Clinical Trials: The Experience of PRION-1.
Large clinical trials including patients with uncommon diseases involve assessors in different geographical locations, resulting in considerable inter-rater variability in assessment scores. As video recordings of examinations, which can be individually rated, may eliminate such variability, we measured the agreement between a single video rater and multiple examining physicians in the context of PRION-1, a clinical trial of the antimalarial drug quinacrine in human prion diseases. We analysed a 43-component neurocognitive assessment battery, on 101 patients with Creutzfeldt-Jakob disease, focusing on the correlation and agreement between examining physicians and a single video rater. In total, 335 videos of examinations of 101 patients who were video-recorded over the 4-year trial period were assessed. For neurocognitive examination, inter-observer concordance was generally excellent. Highly visual neurological examination domains (e.g. finger-nose-finger assessment of ataxia) had good inter-rater correlation, whereas those dependent on non-visual clues (e.g. power or reflexes) correlated poorly. Some non-visual neurological domains were surprisingly concordant, such as limb muscle tone. Cognitive assessments and selected neurological domains can be practically and accurately recorded in a clinical trial using video rating. Video recording of examinations is a valuable addition to any trial provided appropriate selection of assessment instruments is used and rigorous training of assessors is undertaken.


SU-E-T-134: Patient Specific Quality Assurance of RapidArc Pre Treatment Plans Using Semiflex 0.125 Cc Ionization Chamber.
To evaluate the Patient specific pre-treatment quality assurance for hundred RapidArc plans using semiflex (0.125cc) ionization chambers. Absolute point dose were measured for head and neck, thorax and abdomen cases using semiflex (0.125 cc) ionization chamber. Verification plan was created for each treatment plan in eclipse 8.6 treatment planning system with the semiflex ionization chamber and the octavius phantom. Measurements were performed on a Varian Clinac2100C/D linear accelerator equipped with a millennium 120 leaf collimator. All the results were compared with the fluence measurements using 2D Seven29 ion chamber array combined with octavius phantom. Positive absolute mean dose variation of 0.56 % was observed with thorax cases with a standard deviation (SD) of ± 1.13 between the plans with a range of -1.78% to 2.70%. Negative percentage dose errors were found with head and neck and abdomen cases, with a mean variation of -0.43 % (SD ± 1.50), (range -3.25 % to 2.85 %) and -0.35 % (SD ± 1.48), (range -3.10 % to 2.65 %) for head and neck and abdomen cases respectively. Relative dose measurements with 2D array agreed well with the TPS calculate for all the cases. The maximum percentage value failed in gamma analysis was found to be 4.95, 4.75, and 4.88 for head and neck, thorax, and abdomen cases respectively. In all the cases analysed the percentage dose points failed the gamma criteria was less than 5%. On the basis of the studies performed it can be concluded that the semiflex ionization chamber having a volume of 0.125cc can be used efficiently for measuring the pre-treatment quality assurance of RapidArc plans for all the sites. The results provide an overall accuracy when compared to fluence measurement done using 2D array seven29.


A low-energy-consumption electroactive valveless hydrogel micropump for long-term biomedical applications.
Stimuli-responsive hydrogels have attracted considerable interest in the field of microfluidics due to their ability to transform electrical energy directly into mechanical work through swelling, bending, and other deformations. In particular, electroactive hydrogels hold great promise for biomedical micropumping applications such as implantable drug delivery systems. In such applications, energy consumption rate and durability are key properties. Here, we developed a valveless micropump system that utilizes a hydrogel as the main actuator, and tested its performance over 6 months of continuous operation. The proposed micropump system, powered by a single 1.5 V commercial battery, expended very little energy (less than 750 μWs per stroke) while pumping 0.9 wt% saline solution under a low voltage (less than 1 V), and remained fully functional after 6 months. CFD simulations were conducted to improve the microchannel geometry so as to minimize the backflow caused by the valveless mechanism of the system. Based on the simulation results, an asymmetric geometry and a stop post were introduced to enhance the pumping performance. To demonstrate the feasibility of the proposed system as a drug delivery pump, an anti-cancer drug (adriamycin) was perfused to human breast cancer cells (MCF-7) using the pump. The present study showed that the proposed system can operate continuously for long periods with low energy consumption, powered by a single 1.5 V battery, making it a promising candidate for an implantable drug delivery system.


Culture of Keratinocytes for Transplantation without the Need of Feeder Layer Cells.
Patients with large burn wounds have a limited amount of healthy donor skin. An alternative for the autologous skin graft is transplantation with autologous keratinocytes. Conventionally, the keratinocytes are cultured with mouse feeder layer cells in medium containing fetal calf serum (FCS) to obtain sufficient numbers of cells. These xenobiotic materials can be a potential risk for the patient. The aim of the present study was to investigate if keratinocytes could be expanded in culture without the need of a feeder layer and FCS. Keratinocytes were cultured on tissue culture plastic with or without collagen type IV coating in medium containing Ultroser G (serum substitute) and keratinocyte growth factor (KGF). An in vitro skin equivalent model was used to examine the capacity of these cells to form an epidermis. Keratinocytes in different passages (P2, P4, and P6) and freshly isolated cells were studied. Keratinocytes grown on collagen type IV were able to form an epidermis at higher passage numbers than cells grown in the absence of collagen type IV (P4 and P2, respectively). In both cases the reconstructed epidermis showed an increased expression of Ki-67, SKALP, involucrin, and keratin 17 compared to normal skin. Only 50,000 keratinocytes grown on collagen type IV in P4 were needed to form 1 cm2 epidermis, whereas 150,000 of freshly isolated keratinocytes were necessary. Using this culture technique sufficient numbers of keratinocytes, isolated from 1 cm2 skin, were obtained to cover 400 cm2 of wound surface in 2 weeks. The results show that keratinocytes can be cultured without the need of a fibroblast feeder layer and FCS and that these cells are still able to create a fully differentiated epidermis. This culture technique can be a valuable tool for the treatment of burn wounds and further development of tissue engineered skin.


Emergence of spontaneous anticipatory hand movements in a probabilistic environment.
In this article, we present a novel experimental approach to the study of anticipation in probabilistic cuing. We implemented a modified spatial cuing task in which participants made an anticipatory hand movement toward one of two probabilistic targets while the (x, y)-computer mouse coordinates of their hand movements were sampled. This approach allowed us to tap into anticipatory processes as they occurred, rather than just measuring their behavioral outcome through reaction time to the target. In different conditions, we varied the participants' degree of certainty of the upcoming target position with probabilistic pre-cues. We found that participants initiated spontaneous anticipatory hand movements in all conditions, even when they had no information on the position of the upcoming target. However, participants' hand position immediately before the target was affected by the degree of certainty concerning the target's position. This modulation of anticipatory hand movements emerged rapidly in most participants as they encountered a constant probabilistic relation between a cue and an upcoming target position over the course of the experiment. Finally, we found individual differences in the way anticipatory behavior was modulated with an uncertain/neutral cue. Implications of these findings for probabilistic spatial cuing are discussed.


A randomized double-blind placebo-controlled study of Pu'er tea extract on the regulation of metabolic syndrome.
To explore the regulative efficacy of Pu'er tea () extract on metabolic syndrome. Ninety patients with metabolic syndrome were randomly divided into two groups, the intervention group administered with Pu'er tea extract, and the placebo group with placebo capsules. After 3 months' treatment, body mass index, waist hip ratio, blood lipids, blood sugar, immune and inflammatory index, and oxidation index of the patients with metabolic syndrome were tested and analyzed. In the intervention group, the body mass index, waist-hip ratio, fasting and 2 h postprandial blood glucose, serum total cholesterol, triglycerides, low density lipoprotein and apolipoprotein B-100 all decreased in the patients with metabolic syndrome, and also the high-density lipoprotein level increased and apolipoprotein A-1 showed the tendency to increase. Serum C-reactive protein, tumor necrosis factor-α, and interleukin-6 were decreased in the intervention group. Interleukin-10 level was increased, MDA was decreased and superoxide dismutase was increased. Compared with before treatment and the placebo group, there were significant differences (P<0.05, P<0.01). Pu'er tea demonstrated excellent potential in improving central obesity, adjusting blood lipid, lowering blood sugar, regulating immunity and resisting oxidation. It can adjust the metabolic syndrome of different clinical phenotypes to different degrees, and is ideally fit for early prevention of metabolic syndrome.


Intermittent claudication caused by atherosclerosis in patients aged forty years and younger.
Intermittent claudication may represent a severe limitation in daily life-style for young, active patients and may result in loss of professional career for members of the armed forces. Our recent experience with young patients with atherosclerotic claudication was reviewed to determine whether more liberal use of angioplasty and surgery is warranted in these patients to salvage their military careers and improve their life-styles. A systemic review was made of patients aged 40 years and younger with atherosclerosis who have undergone arteriography at our institution during the past 67 months. From this study, 22 patients were found with the initial complaint of intermittent claudication. Fifteen of these patients (68%) had occlusive disease confined to the iliac arteries, and six (27%) had single-level occlusive disease in a femoropopliteal distribution. Only one patient had multilevel disease. Symptoms of coronary occlusive disease developed in five patients (23%) within 38 months of the onset of claudication, and two of these patients died of myocardial infarctions. Of 19 patients who underwent interventions to correct symptoms of claudication, 16 (85%) had complete relief of symptoms at the end of follow-up, ranging from 6 months to 7 years. Ten of these patients remained symptom free after a single intervention, but six patients had rapid progression of disease and required secondary procedures. These results indicate that a majority of young patients with claudication caused by atherosclerosis can be returned to full active-duty status in the military after angioplasty or surgery. However, some patients may require secondary procedures to relieve recurrent symptoms caused by rapid progression of disease. We believe that an aggressive interventional approach is warranted in the evaluation and treatment of young patients with work-limiting claudication caused by atherosclerosis. These patients are at significant risk of developing premature coronary occlusive disease and should be monitored closely.


Directional atherectomy before stenting versus stenting alone in percutaneous coronary interventions: a meta-analysis.
Plaque debulking before stenting is still controversial. We performed a meta-analysis of 12 randomized and non-randomized trials comparing directional coronary atherectomy (DCA) before stenting versus stenting alone. Angiographic end points were acute gain, late loss and angiographic restenosis rate. Clinical end points were early major adverse cardiac events [MACEs: death, Q-wave myocardial infarction (MI), non-Q-wave MI], late MACEs (death, Q-wave MI) and target lesion revascularization (TLR). Data are expressed as odds ratio (OR) with 95% confidence intervals (CI) or weighted mean difference (WMD) with 95% CI, as appropriate. A total of 1216 patients undergoing DCA before stent and 1484 patients undergoing stent alone have been included. DCA before stent was associated to a better acute gain compared to stenting alone (WMD 0.23, [0.18-0.28]; p<0.0001), to a striking reduction of angiographic restenosis rate (OR of 0.67, [0.54-0.84], p=0.0003) and to a significantly lower rate of late TLR (OR 0.73 [0.59-0.91], p=0.006). Late loss did not differ between the two groups (WMD 0.00 [-0.08 and 0.08], p=0.98). We found a higher rate of early MACEs for the combined approach (OR 1.87 [1.16-3.02], p=0.01), with similar prevalence of late MACEs (OR 0.83 [0.65-1.06], p=0.13). In conclusion, this meta-analysis demonstrates that DCA before stenting is superior to stenting alone with regard to acute angiographic results and TLR with a similar prevalence of late MACEs. The higher prevalence of early MACEs with DCA before stenting, however, is disturbing and probably related to distal embolization.


ECIL guidelines for preventing Pneumocystis jirovecii pneumonia in patients with haematological malignancies and stem cell transplant recipients.
The 5th European Conference on Infections in Leukaemia (ECIL-5) meeting aimed to establish evidence-based recommendations for the prophylaxis of Pneumocystis jirovecii pneumonia (PCP) in non-HIV-infected patients with an underlying haematological condition, including allogeneic HSCT recipients. Recommendations were based on the grading system of the IDSA. Trimethoprim/sulfamethoxazole given 2-3 times weekly is the drug of choice for the primary prophylaxis of PCP in adults ( A-II: ) and children ( A-I: ) and should be given during the entire period at risk. Recent data indicate that children may benefit equally from a once-weekly regimen ( B-II: ). All other drugs, including pentamidine, atovaquone and dapsone, are considered second-line alternatives when trimethoprim/sulfamethoxazole is poorly tolerated or contraindicated. The main indications of PCP prophylaxis are ALL, allogeneic HSCT, treatment with alemtuzumab, fludarabine/cyclophosphamide/rituximab combinations, >4 weeks of treatment with corticosteroids and well-defined primary immune deficiencies in children. Additional indications are proposed depending on the treatment regimen.


Simultaneous inhibition of T helper 2 and T regulatory cell differentiation by small molecules enhances Bacillus Calmette-Guerin vaccine efficacy against tuberculosis.
Tuberculosis affects nine million individuals and kills almost two million people every year. The only vaccine available, Bacillus Calmette-Guerin (BCG), has been used since its inception in 1921. Although BCG induces host-protective T helper 1 (Th1) cell immune responses, which play a central role in host protection, its efficacy is unsatisfactory, suggesting that additional methods to enhance protective immune responses are needed. Recently we have shown that simultaneous inhibition of Th2 cells and Tregs by using the pharmacological inhibitors suplatast tosylate and D4476, respectively, dramatically enhances Mycobacterium tuberculosis clearance and induces superior Th1 responses. Here we show that treatment with these two drugs during BCG vaccination dramatically improves vaccine efficacy. Furthermore, we demonstrate that these drugs induce a shift in the development of T cell memory, favoring central memory T (Tcm) cell responses over effector memory T (Tem) cell responses. Collectively, our findings provide evidence that simultaneous inhibition of Th2 cells and Tregs during BCG vaccination promotes vaccine efficacy.


European Respiratory Society guideline on long-term management of children with bronchopulmonary dysplasia.
This document provides recommendations for monitoring and treatment of children in whom bronchopulmonary dysplasia (BPD) has been established and who have been discharged from the hospital, or who were >36 weeks of postmenstrual age. The guideline was based on predefined Population, Intervention, Comparison and Outcomes (PICO) questions relevant for clinical care, a systematic review of the literature and assessment of the evidence using the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) approach. After considering the balance of desirable (benefits) and undesirable (burden, adverse effects) consequences of the intervention, the certainty of the evidence, and values, the task force made conditional recommendations for monitoring and treatment of BPD based on very low to low quality of evidence. We suggest monitoring with lung imaging using ionising radiation in a subgroup only, for example severe BPD or recurrent hospitalisations, and monitoring with lung function in all children. We suggest to give individual advice to parents regarding daycare attendance. With regards to treatment, we suggest the use of bronchodilators in a subgroup only, for example asthma-like symptoms, or reversibility in lung function; no treatment with inhaled or systemic corticosteroids; natural weaning of diuretics by the relative decrease in dose with increasing weight gain if diuretics are started in the neonatal period; and treatment with supplemental oxygen with a saturation target range of 90-95%. A multidisciplinary approach for children with established severe BPD after the neonatal period into adulthood is preferable. These recommendations should be considered until new and urgently needed evidence becomes available.


Hereditary Angioedema Due to C1-Inhibitor Deficiency in Romania: First National Study, Diagnostic and Treatment Challenges.
Hereditary angioedema (HAE) is a rare genetic potentially life-threatening disease characterized by episodic non-pruritic subcutaneous and submucosal edema attacks in different parts of the body. To assess the status of Romanian HAE patients after the recent introduction of a new therapy through a nationwide program. This cross-sectional observational study included patients from the Romanian HAE Registry. The study included 84 patients with HAE type I (91.7%) and type II (8.3%). The mean delay in diagnosis was 2.4 years in children and 16.7 years in adults (p=0.019). Stress and tiredness were the most frequent trigger factors. The majority of the HAE episodes involved subcutaneous (89.3%), abdominal (77.4%), genital (51.2%), facial (41.7%), and laryngeal (39.3%) symptoms during the preceding 12 months. One or several misdiagnoses were reported in 83.33% patients and 44.1 % of the patients were subjected to or proposed unnecessary surgery during abdominal episodes. Plasma-derived C1-INH (pdC1-INH) and recombinant C1-INH (rhC1-INH) were respectively used in 10 (11.9%) and 13 (15.5%) of the HAE patients for life-threatening attacks over the past 12 months. Forty-three (51.19%) patients practiced home treatment with subcutaneous injection of the bradykinin B2-receptor antagonist for acute HAE attacks. The significantly lower delay observed in children suggests an improvement in the awareness of C1-INH-HAE among physicians in recent years. The management of HAE in Romania has been somewhat enhanced as the majority of HAE patients have recently gained access to pdC1-INH, rhC1-INH, and bradykinin B2-receptor antagonist.


Biotransformation of the organophosphorus insecticides parathion and methyl parathion in male and female rat livers perfused in situ.
Although numerous previous reports have characterized the mammalian biotransformation of the organophosphorus insecticides parathion and methyl parathion, questions still remain regarding the toxicological significance of certain metabolic pathways in vivo. The present study utilized rat liver perfusions in order to better characterize the hepatic biotransformation of parathion and methyl parathion in intact liver. Single-pass liver perfusions with parathion and methyl parathion over a range of perfusate concentrations of 10-80 microM resulted in the appearance of paraoxon and methyl paraoxon, respectively, in effluent. Furthermore, rat blood did not have the capacity to prevent transport of paraoxon and methyl paraoxon to extrahepatic tissues, suggesting that oxon produced hepatically can distribute to extrahepatic tissues. In addition, striking sex differences were noted in the metabolite profile of parathion and methyl parathion in perfused livers. However, these differences could not account for the observation that females are more susceptible to parathion, but less susceptible to methyl parathion, compared to males. And finally, S-methyl glutathione or S-p-nitrophenyl glutathione could not be detected in effluent or bile of livers from either sex perfused with methyl parathion, suggesting that glutathione-dependent detoxification of this insecticide does not occur to any significant degree in intact rat liver.


Normal patterns of melatonin levels in the pineal gland and body fluids of humans and experimental animals.
The normal 24 hours patterns of melatonin were described in experimental animals and man. The rhythms are generated by neuronal activity originating in the suprachiasmatic nuclei of the hypothalamus; the activity of these nuclei are synchronized by the prevailing light:dark environment as perceived by the retinas. As a consequence of this synchronization, pineal melatonin levels are always high at night and low during the day, irrespective of whether the animal being studied is nocturnally or diurnally active. The rhythmic production of melatonin in the pineal gland is interrupted by the exposure of animals, or man, to light during the normal dark period. Several factors may change the 24 hours pattern of melatonin production. Because daylength changes throughout the year there are seasonal effects on the melatonin rhythm. Also, age is a major factor in determining the ability of the pineal gland to metabolize serotonin to melatonin. In advanced age pineal melatonin production is severely limited in both the human and in experimental animals. A variety of endocrine manipulations have been tested in terms of their impact on the ability of the pineal to produce melatonin. The majority of these have been rather innocuous although hypophysectomy does significantly curtail the synthesis of melatonin. To date, melatonin cycles have been usually described in rodents maintained under controlled environmental conditions in the laboratory; it can only be assumed that these rhythms are similar in animals in their natural habitat.


The fitness value of information.
Communication and information are central concepts in evolutionary biology. In fact, it is hard to find an area of biology where these concepts are not used. However, quantifying the information transferred in biological interactions has been difficult. How much information is transferred when the first spring rainfall hits a dormant seed, or when a chick begs for food from its parent? One measure that is commonly used in such cases is fitness value: by how much, on average, an individual's fitness would increase if it behaved optimally with the new information, compared to its average fitness without the information. Another measure, often used to describe neural responses to sensory stimuli, is the mutual information-a measure of reduction in uncertainty, as introduced by Shannon in communication theory. However, mutual information has generally not been considered to be an appropriate measure for describing developmental or behavioral responses at the organismal level, because it is blind to function; it does not distinguish between relevant and irrelevant information. In this paper we show that there is in fact a surprisingly tight connection between these two measures in the important context of evolution in an uncertain environment. In this case, a useful measure of fitness benefit is the increase in the long-term growth rate, or the fold increase in number of surviving lineages. We show that in many cases the fitness value of a developmental cue, when measured this way, is exactly equal to the reduction in uncertainty about the environment, as described by the mutual information.


Triggers of self-conscious emotions in the sexually transmitted infection testing process.
Self-conscious emotions (shame, guilt and embarrassment) are part of many individuals' experiences of seeking STI testing. These emotions can have negative impacts on individuals' interpretations of the STI testing process, their willingness to seek treatment and their willingness to inform sexual partners in light of positive STI diagnoses. Because of these impacts, researchers have called for more work to be completed on the connections between shame, guilt, embarrassment and STI testing. We examine the specific events in the STI testing process that trigger self-conscious emotions in young adults who seek STI testing; and to understand what it is about these events that triggers these emotions.Semi-structured interviews with 30 adults (21 women, 9 men) in the Republic of Ireland. Seven specific triggers of self-conscious emotions were identified. These were: having unprotected sex, associated with the initial reason for seeking STI testing; talking to partners and peers about the intention to seek STI testing; the experience of accessing STI testing facilities and sitting in clinic waiting rooms; negative interactions with healthcare professionals; receiving a positive diagnosis of an STI; having to notify sexual partners in light of a positive STI diagnosis; and accessing healthcare settings for treatment for an STI. Self-conscious emotions were triggered in each case by a perceived threat to respondents' social identities. There are multiple triggers of self-conscious emotions in the STI testing process, ranging from the initial decision to seek testing, right through to the experience of accessing treatment. The role of self-conscious emotions needs to be considered in each component of service design from health promotion approaches, through facility layout to the training of all professionals involved in the STI testing process.


Breast oedema following free flap breast reconstruction.
Breast oedema causes significant morbidity and is historically difficult to quantify. The aim of this study was to identify changes in breast tissue water content from pre-operative levels in the native breast to post-operative levels in mastectomy skin flaps and free flaps in the reconstructed breast. One hundred patients undergoing unilateral mastectomy and immediate free flap breast reconstruction were examined pre-operatively and at three post-operative appointments. A validated moisture meter was used to record dermal water percentages of each breast quadrant and areola in both breasts pre-operatively, then four quadrants of both breasts plus the unaffected areola and free flap at each post-operative review. Native skin of the reconstructed breast showed significant, persistent increase in MWC from 45.6% ± 0.5% to 72.8% ± 0.9% at 1st follow up (p < 0.001), decreasing only to 67.6% ± 0.8% by 3rd follow up. There was a marked difference (p < 0.001) in the mean water content (MWC) of the initial free flap (39.7% ± 0.6%) compared to 61.8% ± 1.7% at 1st follow up, then 55.1% ± 1.4% at 2nd and 53.7% ± 1.3% at 3rd follow ups. The unaffected breast showed a small but significant increase in MWC of all quadrants at subsequent follow up (greatest difference 3.1% at 1st follow up). This patient group demonstrates significant, persistent oedema of the reconstructed breast, which can be monitored using a non-invasive moisture meter.


Receptor-interacting protein 140 binds c-Jun and inhibits estradiol-induced activator protein-1 activity by reversing glucocorticoid receptor-interacting protein 1 effect.
In the presence of estradiol, estrogen receptor-alpha (ERalpha) increases transcription triggered by activator protein-1 (AP-1). We have previously shown that induction is mediated by the direct interaction between c-Jun and ERalpha, which stabilizes a multiprotein complex containing the coactivator GRIP1 (glucocorticoid receptor interacting protein 1). The effect of receptor-interacting protein 140 (RIP140) in this regulation was assessed in the present study. We report that overexpression of RIP140 inhibits estradiol-induced AP-1-dependent transcription in a dose-dependent manner. Inhibition is not affected by trichostatin A, suggesting that histone deacetylase recruitment is not implicated. RIP140, which binds Jun proteins in pull-down assays and in intact cells, as shown by coimmunoprecipitation analysis and a mammalian one-hybrid system, participates in a multiprotein complex containing c-Jun and ERalpha. Moreover, the negative effect of RIP140 on AP-1-mediated transcription is relieved by GRIP1 overexpression and, conversely, RIP140 inhibits the stimulatory effect of GRIP1. The two cofactors compete for binding to c-Jun and ERalpha both in vitro and in intact cells, and GRIP1 interaction with both ERalpha and c-Jun is required for an efficient competition. These overall results suggest that the ratio between RIP140 and GRIP1 could determine, as proposed for hormone response element-mediated responses, the efficacy of estradiol in stimulating transcription of genes under AP-1 control.


HLA class I-minor histocompatibility antigen tetramers select cytotoxic T cells with high avidity to the natural ligand.
Cytotoxic T cells specific for the hematopoietic system-restricted minor histocompatibility antigens HA-1 and HA-2 are potential tools for the treatment of relapsed leukemia after minor histocompatibility antigen mismatched bone marrow transplantation. HA-1/HA-2-specific cytotoxic T cells with strong cytotoxic activity against HA-1/HA-2 positive target cells can be generated in vitro using HA-1 and HA-2 peptide-pulsed dendritic cells as antigen presenting cells. We used HLA-A2 HA-1/HA-2 tetramers (HA-1(A2)/HA-2(A2) tetramers) to monitor the in vitro generation of HA-1- or HA-2-specific cytotoxic T cells. We show that the intensity of the tetramer-staining of the HA-1/HA-2-specific cytotoxic T cells strongly correlates with their capability to recognize mHag positive target cells. The bright tetramer-staining cytotoxic T cells lyse target cells expressing the natural ligand. The dim tetramer-staining cytotoxic T cells fail to lyse natural ligand positive target cells and lyse peptide-pulsed target cells only. The frequency of bright tetramer-staining, high avidity minor histocompatibility antigen-specific CTLs increases significantly upon appropriate antigen-specific restimulations. Our results demonstrate that HLA class I-minor histocompatibility antigen tetramers are useful tools for monitoring and selection of high avidity HA-1- and HA-2-specific cytotoxic T cells for adoptive immunotherapy.


Nurses' health beliefs about paper face masks in Japan, Australia and China: a qualitative descriptive study.
To explore the health beliefs of clinical and academic nurses from Japan, Australia and China regarding wearing paper masks to protect themselves and others, and to identify differences in participants' health beliefs regarding masks. The correct use of face masks and consensus among health professionals across the globe is essential for containing pandemics, and nurses need to act according to policy to protect themselves, educate the public and preserve resources for frontline health workers. Paper masks are worn by health professionals and the general public to avoid the transmission of respiratory infections, such as COVID-19, but there appear to be differences in health beliefs of nurses within and between countries regarding these. This qualitative descriptive study used content analysis with a framework approach. There were major differences in nurse participants' beliefs between and within countries, including how nurses use paper masks and their understanding of their efficacy. In addition, there were cultural differences in the way that nurses use masks in their daily lives and nursing practice contexts. Nurses from different working environments, countries and areas of practice hold a variety of health beliefs about mask wearing at the personal and professional level. The COVID-19 pandemic has sparked much discussion about the critical importance of masks for the safety of health professionals, and there has been considerable discussion and disagreement about health policies regarding mask use by the general public. Improper use of masks may have a role in creating mask shortages or transmitting infections. An evidence-based global policy on mask use for respiratory illnesses for health professionals, including nurses, and the general public needs to be adopted and supported by a wide-reaching education campaign.


Current safety practices relating to I-131 administration for diseases of the thyroid: a survey of physicians and allied practitioners.
There is little information about the individual safety instructions provided by healthcare professionals to patients receiving radioactive iodine (I-131) therapy for the treatment of benign and malignant thyroid disorders or about whether these instructions are consistent across medical specialties. Currently, no national guidelines exist to standardize safety instructions related to I-131 administration. Here, we examine the spectrum of I-131 safety practices in contemporary use. Members of major societies of physicians and allied specialists who treat patients with thyroid disorders were invited to complete a 27-question online survey about safety practices related to I-131 administration. Data from questionnaires were analyzed by type of safety recommendation and grouped according to provider specialty and geographic location. A total of 311 endocrinologists, surgeons, nuclear medicine radiologists, and allied health professionals completed questionnaires. They indicated that patients often receive instruction from more than one treating specialist. The decision to hospitalize a patient for treatment and the length of stay were determined by the patient's social situation and the dose of I-131 administered. Starting at I-131 doses between 259 and 1073 MBq (7 and 29 mCi), over 60% of respondents advised avoiding contact with children, sexual activity, and breastfeeding, with the latter recommendation continuing beyond 48 hours after treatment. Personal hygiene, laundry, and meal preparation precautions varied across respondents. Over 90% of respondents used serum or urine testing to screen for pregnancy status. Precautions to delay parenthood were given more often to female than male patients (90% vs. 60%), with a minimum recommended delay of 6 months. About 20% of respondents considered insurance coverage as a factor in selecting outpatient versus inpatient I-131 therapy, and this consideration varied geographically. A wide variety of safety recommendations are given to patients who receive I-131. To our knowledge, this survey represents the first organized inquiry into safety practices related to I-131 administration. The diversity of responses suggests an opportunity for multispecialty collaboration in defining more uniform recommendations for patient safety instructions during and after I-131 treatment.


Responsiveness of gonadotropin secretion to infusion of an opiate-receptor antagonist in hypogonadotropic individuals.
We tested the hypothesis that suppressive effects of endogenous opiate substances are involved in certain hypogonadotropic states. For this purpose, we studied gonadotropin secretion in idiopathic hypopituitarism (five children), constitutionally delayed adolescence (five boys), and Kallmann's syndrome (three men). Endogenous opiate pathways were antagonized by the iv infusion of naloxone hydrochloride at a dose previously shown to elicit a prompt and significant increase in serum levels of LH in normal men. Under these conditions, naloxone did not increase serially sampled serum concentrations or mean urinary levels of LH: or FSH in eight patients with idiopathic hypopituitarism or Kallmann's syndrome. Gonadotropin concentrations in four of five patients with constitutional delay of adolescence also were unaffected. In one boy with clinical and biochemical indices of late pubertal development, naloxone elicited a significant increase in LH levels in blood and urine, similar to the pattern observed in normal men. In contrast to results in experimental animals, naloxone did not suppress serum PRL concentration significantly in any subject. These observations suggest that: 1) endogenous opiate mechanisms are unlikely to constitute a principal factor in maintaining hypogonadotropism in idiopathic hypopituitarism, delayed adolescence, or Kallmann's syndrome, at least acutely; 2) endogenous opiate mechanisms also cannot be implicated in the acute regulation or PRL secretion in children; and 3) the capability of adult men, but not early pubertal boys, to respond with increased gonadotropin secretion during inhibition of opiate receptors suggests that maturation of the opiate-related neuroendocrine system occurs during the course of sexual development in the human.


Evidence for high levels of vertical transmission in Toxoplasma gondii.
Toxoplasma gondii is a highly ubiquitous and prevalent parasite. Despite the cat being the only definitive host, it is found in almost all geographical areas and warm blooded animals. Three routes of transmission are recognised: ingestion of oocysts shed by the cat, carnivory and congenital transmission. In natural populations, it is difficult to establish the relative importance of these routes. This paper reviews recent work in our laboratory which suggests that congenital transmission may be much more important than previously thought. Using PCR detection of the parasite, studies in sheep show that congenital transmission may occur in as many as 66% of pregnancies. Furthermore, in families of sheep on the same farm, exposed to the same sources of oocysts, significant divergent prevalences of Toxoplasma infection and abortion are found between different families. The data suggest that breeding from infected ewes increases the risk of subsequent abortion and infection in lambs. Congenital transmission rates in a natural population of mice were found to be 75%. Interestingly, congenital transmission rates in humans were measured at 19.8%. The results presented in these studies differ from those of other published studies and suggest that vertical transmission may be much more important than previously thought.


An introduction to instrumental variables analysis: part 1.
There are several examples in the medical literature where the associations of treatment effects predicted by observational studies have been refuted by evidence from subsequent large-scale randomised trials. This is because of the fact that non-experimental studies are subject to confounding - and confounding cannot be entirely eliminated even if all known confounders have been measured in the study as there may be unknown confounders. The aim of this 2-part methodological primer is to introduce an emerging methodology for estimating treatment effects using observational data in the absence of good randomised evidence known as the method of instrumental variables.


Mitochondrial targeted coenzyme Q, superoxide, and fuel selectivity in endothelial cells.
Previously, we reported that the "antioxidant" compound "mitoQ" (mitochondrial-targeted ubiquinol/ubiquinone) actually increased superoxide production by bovine aortic endothelial (BAE) cell mitochondria incubated with complex I but not complex II substrates. To further define the site of action of the targeted coenzyme Q compound, we extended these studies to include different substrate and inhibitor conditions. In addition, we assessed the effects of mitoquinone on mitochondrial respiration, measured respiration and mitochondrial membrane potential in intact cells, and tested the intriguing hypothesis that mitoquinone might impart fuel selectivity in intact BAE cells. In mitochondria respiring on differing concentrations of complex I substrates, mitoquinone and rotenone had interactive effects on ROS consistent with redox cycling at multiple sites within complex I. Mitoquinone increased respiration in isolated mitochondria respiring on complex I but not complex II substrates. Mitoquinone also increased oxygen consumption by intact BAE cells. Moreover, when added to intact cells at 50 to 1000 nM, mitoquinone increased glucose oxidation and reduced fat oxidation, at doses that did not alter membrane potential or induce cell toxicity. Although high dose mitoquinone reduced mitochondrial membrane potential, the positively charged mitochondrial-targeted cation, decyltriphenylphosphonium (mitoquinone without the coenzyme Q moiety), decreased membrane potential more than mitoquinone, but did not alter fuel selectivity. Therefore, non-specific effects of the positive charge were not responsible and the quinone moiety is required for altered nutrient selectivity. In summary, the interactive effects of mitoquinone and rotenone are consistent with redox cycling at more than one site within complex I. In addition, mitoquinone has substrate dependent effects on mitochondrial respiration, increases repiration by intact cells, and alters fuel selectivity favoring glucose over fatty acid oxidation at the intact cell level.


Cartilage transplantation techniques for talar cartilage lesions.
Talar articular cartilage is known to differ significantly from knee cartilage. Even so, recommendations for the treatment of talar cartilage lesions have been based on strategies for the knee. Arthroscopic management of osteochondral lesions of the talus is well documented. Results have been favorable with reparative techniques such as débridement with curettage and débridement with drilling, whether undertaken via early open techniques or more recent arthroscopic procedures. Salvage of failed reparative techniques is controversial. Early efforts to salvage failed débridement focused on osteochondral allografts and autografts that used the knee as a donor site. Results of these restorative techniques have been favorable, but concerns have been raised regarding knee donor site morbidity, the use of malleolar osteotomy, and incomplete restoration of the talar articular surface. More recent restorative techniques developed for the knee have been adapted for the ankle, such as autologous chondrocyte implantation and matrix-induced autologous chondrocyte implantation.


Paw-reaching, sensorimotor, and rotational behavior after brain infarction in rats.
Functional tests that are stable and consistent over time are an advantage for long-term evaluation of treatment in experimental stroke research. Because little information on this subject is available in rodents with focal cerebral ischemia, we investigated the outcome of three behavioral tests for a period of 3 months after the insult. Spontaneously hypertensive rats were sham-operated (n = 27) or underwent an occlusion (n = 36) of the right middle cerebral artery. Before surgery all rats were tested for amphetamine-induced rotational behavior, and half of the rats were trained in a paw-reaching task. One, 2, and 3 months after surgery the tests were repeated, together with a test for sensorimotor function. Infarct size was measured morphometrically. In the lesion group, total hemisphere area was reduced by 22%, caudate putamen by 47%, and the thalamus by 24%. Contralateral to the lesion, paw-reaching was highly impaired, regardless of whether or not the rats had been pretrained, and lesion size correlated significantly to paw-reach performance. Ipsilateral rotation increased and sensorimotor function recovered with time in infarcted rats. In contrast to amphetamine-induced rotation and sensorimotor behavior, the paw-reaching test provides a stable behavioral parameter after a middle cerebral artery occlusion. Moreover, the lesion-induced deficit in paw-reaching is highly correlated to the extent of the infarct, suggesting that this test is useful in evaluating treatment effects for a longer period of time.


Social networks and cooperation in hunter-gatherers.
Social networks show striking structural regularities, and both theory and evidence suggest that networks may have facilitated the development of large-scale cooperation in humans. Here, we characterize the social networks of the Hadza, a population of hunter-gatherers in Tanzania. We show that Hadza networks have important properties also seen in modernized social networks, including a skewed degree distribution, degree assortativity, transitivity, reciprocity, geographic decay and homophily. We demonstrate that Hadza camps exhibit high between-group and low within-group variation in public goods game donations. Network ties are also more likely between people who give the same amount, and the similarity in cooperative behaviour extends up to two degrees of separation. Social distance appears to be as important as genetic relatedness and physical proximity in explaining assortativity in cooperation. Our results suggest that certain elements of social network structure may have been present at an early point in human history. Also, early humans may have formed ties with both kin and non-kin, based in part on their tendency to cooperate. Social networks may thus have contributed to the emergence of cooperation.


The enhancement of retinal adhesiveness by ouabain appears to involve cellular edema.
Our previous work showed that ouabain increases retinal adhesion to the retinal pigment epithelium, but the mechanism of this effect has been unclear. In this study, we have found that the ouabain-induced increase in retinal adhesiveness was blocked reversibly by removing sodium from the medium or by exposure to hyperosmotic solution. Hyposmotic solution slowed down the postmortem failure of adhesion. Tissue edema could not be documented histologically, but retinal tissue water content increased by 2.9% after exposure to ouabain. These data suggest that the ouabain effect involves cellular edema, which may enhance adhesion by tightening the interdigitation between photoreceptor outer segments and retinal pigment epithelial microvilli.


"They're Not Going to Do Nothing for Me": Research Participants' Attitudes towards Elective Genetic Counseling.
As applications of genomic sequencing have expanded, offering genetic counseling support to all patients is arguably no longer practical. Additionally, whether individuals desire and value genetic counseling services for genomic screening is unclear. We offered elective genetic counseling to 5110 individuals prior to undergoing sequencing and 2310 participants who received neutral results to assess demand. A total of 0.2% of the study participants accessed genetic counseling services prior to sequencing, and 0.3% reached out after receiving neutral results. We later conducted 50 interviews with participants to understand why they did not access these services. Many interviewees did not recall the availability of genetic counseling and were unfamiliar with the profession. Interviewees described not needing counseling before sequencing because they understood the study and felt that they could cope with any result. Counseling was considered equally unnecessary after learning neutral results. Although the participants had questions about their results, they did not feel that speaking with a genetic counselor would be helpful. Genomic screening efforts that employ opt-in models of genetic counseling may need to clarify the potential value of genetic counseling support from the outset and feature genetic counseling services more prominently in program materials.


Cross-reactivity between avian influenza A (H7N9) virus and divergent H7 subtypic- and heterosubtypic influenza A viruses.
The number of human avian H7N9 influenza infections has been increasing in China. Understanding their antigenic and serologic relationships is crucial for developing diagnostic tools and vaccines. Here, we evaluated the cross-reactivities and neutralizing activities among H7 subtype influenza viruses and between H7N9 and heterosubtype influenza A viruses. We found strong cross-reactivities between H7N9 and divergent H7 subtypic viruses, including H7N2, H7N3, and H7N7. Antisera against H7N2, H7N3, and H7N7 could also effectively neutralize two distinct H7N9 strains. Two-way cross-reactivities exist within group 2, including H3 and H4, whereas one-way cross-reactivities were found across other groups, including H1, H10, H9, and H13. Our data indicate that the hemaglutinins from divergent H7 subtypes may facilitate the development of vaccines for distinct H7N9 infections. Moreover, serologic diagnoses for H7N9 infections need to consider possible interference from the cross-reactivity of H7N9 with other subtype influenza viruses.


Is air pollution a risk factor for rheumatoid arthritis?
Rheumatoid arthritis is a chronic inflammatory debilitating disease triggered by a complex interaction involving genetic and environmental factors. Active smoking and occupational exposures such as silica increase its risk, suggesting that initial inflammation and generation of rheumatoid arthritis-related autoantibodies in the lungs may precede the clinical disease. This hypothesis paved the way to epidemiological studies investigating air pollution as a potential determinant of rheumatoid arthritis. Studies designed for epidemiology of rheumatoid arthritis found a link between traffic, a surrogate of air pollution, and this disease. Furthermore, a small case-control study recently found an association between wood smoke exposure and anticyclic citrullinated protein/peptide antibody in sera of patients presenting wood-smoke-related chronic obstructive pulmonary disease. However, reports addressing impact of specific pollutants on rheumatoid arthritis incidence and severity across populations are somewhat conflicting. In addition to the link reported between other systemic autoimmune rheumatic diseases and particulate matters/gaseous pollutants, experimental observation of exacerbated rheumatoid arthritis incidence and severity in mice models of collagen-induced arthritis after diesel exhaust particles exposure as well as hypovitaminosis D-related autoimmunity can help understand the role of air pollution in rheumatoid arthritis. All these considerations highlight the necessity to extend high quality epidemiological researches investigating different sources of atmospheric pollution across populations and particularly in low-and-middle countries, in order to further explore the biological plausibility of air pollution's effect in the pathogenesis of rheumatoid arthritis. This should be attempted to better inform policies aiming to reduce the burden of rheumatoid arthritis.