Site-specific association with host and viral chromatin by Kaposi's sarcoma-associated herpesvirus LANA and its reversal during lytic reactivation.
Latency-associated nuclear antigen (LANA), a multifunctional protein expressed by the Kaposi sarcoma-associated herpesvirus (KSHV) in latently infected cells, is required for stable maintenance of the viral episome. This is mediated by two interactions: LANA binds to specific sequences (LBS1 and LBS2) on viral DNA and also engages host histones, tethering the viral genome to host chromosomes in mitosis. LANA has also been suggested to affect host gene expression, but both the mechanism(s) and role of this dysregulation in KSHV biology remain unclear. Here, we have examined LANA interactions with host chromatin on a genome-wide scale using chromatin immunoprecipitation with high-throughput sequencing (ChIP-seq) and show that LANA predominantly targets human genes near their transcriptional start sites (TSSs). These host LANA-binding sites are generally found within transcriptionally active promoters and display striking overrepresentation of a consensus DNA sequence virtually identical to the LANA-binding site 1 (LBS1) motif in KSHV DNA. Comparison of the ChIP-seq profile with whole-transcriptome (high-throughput sequencing of RNA transcripts [RNA-seq]) data reveals that few of the genes that are differentially regulated in latent infection are occupied by LANA at their promoters. This suggests that direct LANA binding to promoters is not the prime determinant of altered host transcription in KSHV-infected cells. Most surprisingly, the association of LANA to both host and viral DNA is strongly disrupted during the lytic cycle of KSHV. This disruption can be prevented by the inhibition of viral DNA synthesis, suggesting the existence of novel and potent regulatory mechanisms linked to either viral DNA replication or late gene expression. Here, we employ complementary genome-wide analyses to evaluate the distribution of the highly abundant latency-associated nuclear antigen, LANA, on the host genome and its impact on host gene expression during KSHV latent infection. Combined, ChIP-seq and RNA-seq reveal that LANA accumulates at active gene promoters that harbor specific short DNA sequences that are highly reminiscent of its cognate binding sites in the virus genome. Unexpectedly, we found that such association does not lead to remodeling of global host transcription during latency. We also report for the first time that LANA's ability to bind host and viral chromatin is highly dynamic and is disrupted in cells undergoing an extensive lytic reactivation. This therefore suggests that the association of LANA to chromatin during a productive infection cycle is controlled by a new regulatory mechanism.


An Anti-tumorigenic Role of the Warburg Effect at Emergence of Transformed Cells.
The Warburg effect is one of the hallmarks of cancer cells, characterized by enhanced aerobic glycolysis. Despite intense research efforts, its functional relevance or biological significance to facilitate tumor progression is still debatable. Hence the question persists when and how the Warburg effect contributes to carcinogenesis. Especially, the role of metabolic changes at a very early stage of tumorigenesis has received relatively little attention, and how aerobic glycolysis impacts tumor incidence remains largely unknown. Here we discuss a novel paradigm for the effect of the Warburg effect that provides a suppressive role in oncogenesis.Key words: Warburg effect, aerobic glycolysis, cell competition, EDAC.


Evaluation of potential early markers of chorioamnionitis associated with preterm premature ruptured membranes.
Optimal expectant management of preterm premature rupture of membranes (PROM) requires the early detection of chorioamnionitis. To date, however, no universally sensitive and specific marker for chorioamnionitis has been identified. Recently, the serial determination of plasma fibronectin, antithrombin, and prekallikrein has been reported to facilitate the early detection of sepsis in critically ill neonates and adult surgical patients. A cross-sectional study was undertaken to determine if plasma levels of these markers change significantly in patients with overt chorioamnionitis following expectant management of preterm PROM. Plasma levels of fibronectin and prekallikrein were not significantly different between the study (30 patients with overt chorioamnionitis following preterm PROM) and control (30 undelivered patients without antenatal complication matched for gestational age) groups. Antithrombin levels were significantly lower in the study group (p less than 0.05), but the magnitude of the difference (102% versus 94%) is not likely to be of clinical significance. We conclude that determination of plasma levels of fibronectin, prekallikrein, and antithrombin is not likely to aid in the early detection of chorioamnionitis in the setting of preterm (PROM).


The prevalence, serogroup distribution and risk factors of meningococcal carriage in adolescents and young adults in Turkey.
The serogroup epidemiology of invasive meningococcal disease (IMD), which varies considerably by geographic region and immunization schedule, changes continuously. Meningococcal carriage data are crucial for assessing IMD epidemiology and designing f potential vaccination strategies. Meningococcal seroepidemiology in Turkey differs from that in other countries: serogroups W and B are the predominant strains for IMD during childhood, whereas no serogroup C cases were identified over the last 10 y and no adolescent peak for IMD was found. There is a lack of data on meningococcal carriage that represents the whole population. The aims of this multicenter study (12 cities in Turkey) were to evaluate the prevalence of Neisseria meningitidis carriage, the serogroup distribution and the related risk factors (educational status, living in a dormitory or student house, being a household contact with Hajj pilgrims, smoking, completion of military service, attending bars/clubs) in 1518 adolescents and young adults aged 10-24 y. The presence of N. meningitidis DNA was tested, and a serogroup analysis was performed using polymerase chain reaction. The overall meningococcal carriage rate was 6.3% (n = 96) in the study population. A serogroup distribution of the 96 N. meningitidis strains isolated from the nasopharyngeal specimens revealed serogroup A in 5 specimens (5.2%), serogroup B in 9 specimens (9.4%), serogroup W in 64 specimens (66.6%), and serogroup Y in 4 specimens (4.2%); 14 were classified as non-grouped (14.4%). No serogroup C cases were detected. The nasopharyngeal meningococcal carriage rate was 5% in the 10-14 age group, 6.4% in the 15-17 age-group, and 4.7% in the 18-20 age group; the highest carriage rate was found in the 21-24 age group (9.1%), which was significantly higher than those of the other age groups (p < 0.05). The highest carriage rate was found in 17-year-old adolescents (11%). The carriage rate was higher among the participants who had had close contact with Hajj/Umrah pilgrims (p < 0.01) or a history of upper respiratory tract infections over the past 3 months (p < 0.05). The nasopharyngeal carriage rate was 6.3% among adolescents and young adults in Turkey and was similar to the recent rates observed in the same age groups in other countries. The most prevalent serogroup was W, and no serogroup C cases were found. In conclusion, the present study found that meningococcal carriage reaches its peak level by age 17, the highest carriage rate was found in 21 - to 24 - year-olds and the majority of the carriage cases were due to serogroup W. Adolescents and young adult carriers seem to be a potential reservoir for the disease, and further immunization strategies, including adolescent immunization, may play a role in the control of IMD.


Lesbian health care. What a primary care physician needs to know.
Many primary care physicians take care of lesbians and women sexually active with women without being aware of their patients' sexual orientation. These women have unique medical and psychosocial needs that each physician must consider. Lesbian identity or being sexually active exclusively with women influences care in areas such as sexually transmitted diseases, risk of human immunodeficiency virus infection, counseling, cancer risk, screening, parenting, depression, alcohol use, and violence. We review an approach to taking a history with all women that facilitates open, comfortable communication with lesbians. We also review specific medical and psychosocial areas of primary care in which caring for lesbians is different from caring for other women. Further research is needed on lesbian health issues to provide appropriate guidelines to clinicians.


[Fall in aortic diastolic pressure immediately after aortic valve destruction can predict severity of regurgitation through the valve and chronic morphological changes in the left ventricle].
Aortic regurgitation (AR) has been studied experimentally, however these reports did not investigate compensatory mechanisms in AR in relation with the severity of regurgitation. If the severity of AR can be changed voluntarily, we can evaluate the pathophysiological response to AR in wide range. The purposes of this study were 1) to see if the extent of fall in aortic diastolic pressure immediately after the production of AR (delta AoDP) was a reliable predictor of the severity of AR and 2) to evaluate the process of adaptation through acute and chronic stage in moderate AR in the rabbit model. AR was produced by perforating the aortic valves in Japanese white rabbits. First, aortic pressure, volume, weight and wall thickness of the left ventricle were measured. Correlations between delta AoDP and body weight-corrected volume (LVV/BW) and weight (LVW/BW) were studied in 18 rabbits soon after, 19 one week after, 23 four weeks after and 10 eight weeks after the production of AR. delta AoDP was closely correlated with LVV/BW after 1 week (1 week: R = 0.667, p less than 0.01, 4 weeks: R = 0.733, p less than 0.01, 8 weeks: R = 0.757, p less than 0.05) and with LVW/BW after 4 weeks (4 weeks: R = 0.484, p less than 0.05, 8 weeks: R = 0.651, p less than 0.05). The slope of these regression equations increased with time (delta AoDP and LVV/BW: 1 week: 0.0138, 4 weeks: 0.0361, 8 weeks: 0.0577 ml/kg.mmHg; delta AoDP and LVW/BW; 4 weeks: 0.0142 8 weeks: 0.0310 g/kg.mmHg).(ABSTRACT TRUNCATED AT 250 WORDS)


Application of a Colored Multiexposure High Dynamic Range Technique to Radiographic Imaging: An Experimental Trial to Show Feasibility.
The aim of this study was to evaluate the feasibility of applying the high dynamic range (HDR) technique to radiographic imaging to expand the dynamic range of conventional radiographic images using a colored multiexposure approach. An appropriate study object was repeatedly imaged using a range of different imaging parameters using a standard clinical x-ray unit. An underexposed image (acquired at 80 keV), an intermediate exposed image (110 keV), and an overexposed image (140 keV) were chosen and combined to a 32-bit colored HDR image. To display the resulting HDR image on a regular color display with typically 8 bits per channel, the Reinhard tone mapping algorithm was applied. The source images and the resulting HDR image were qualitatively evaluated by 5 independent radiologists with regard to the visibility of the different anatomic structures using a Likert scale (1, not visible, to 5, excellent visibility). Data were presented descriptively. High dynamic range postprocessing was possible without malalignment or image distortion. Application of the Reinhardt algorithm did not cause visible artifacts. Overall, postprocessing time was 7 minutes 10 seconds for the whole process. Visibility of anatomic structure was rated between 1 and 5, depending on the anatomic structure of interest. Most authors rated the HDR image best before individual source images. This experimental trial showed the feasibility of applying the HDR technique to radiographic imaging to expand the dynamic range of conventional radiographic images using a colored multiexposure approach.


Development and testing of hyperpolarized (13)C MR calibrationless parallel imaging.
A calibrationless parallel imaging technique developed previously for (1)H MRI was modified and tested for hyperpolarized (13)C MRI for applications requiring large FOV and high spatial resolution. The technique was demonstrated with both retrospective and prospective under-sampled data acquired in phantom and in vivo rat studies. A 2-fold acceleration was achieved using a 2D symmetric EPI readout equipped with random blips on the phase encode dimension. Reconstructed images showed excellent qualitative agreement with fully sampled data. Further acceleration can be achieved using acquisition schemes that incorporate multi-dimensional under-sampling.


Accelerated secure key distribution based on localized and asymmetric fiber interferometers.
We propose and experimentally demonstrate an approach to generate and distribute secret keys over optical fiber communication infrastructure. Mach-Zehnder interferometers (MZIs) are adopted for key generation by transferring the environmental noise to random optical signals. A novel combination of wideband optical noise and an asymmetric MZI structure enables the secret keys to be securely transmitted and exchanged over public fiber links without being detected. We experimentally demonstrate this system and show reliable performance: keys are generated at the rate of 502 bit/s, and are successfully exchanged between two parties over a 10 km optical fiber with a bit error of ∼ 0.3%. System security analysis is performed by corroborating our experimental findings with simulations. The results show that our system can protect the key distribution under different attacks, attributed to wideband optical noise and asymmetric MZI structures. Compared to the previous schemes based on distributed MZIs, our scheme exploits localized MZI which provides twofold advantages. Firstly, the key generation rate can be increased by a factor of 5.7 at a negligible additional cost. Secondly, the system becomes robust to, in particular, active intrusion attack. The proposed system is a reliable and cost-effective solution for key establishment, and is compatible with the existing optical fiber communication infrastructure.


[A case of Scedosporium apiospermum keratitis confirmed by a molecular genetic method].
A 54-yr-old male, who was treated by chemotherapy for gastric cancer 15 months ago, presented to Yongdong Severance Hospital, Seoul, with complaints of pain in his right eye caused by a foreign body from the ground in the previous week. He had been treated with topical and oral antibacterial in addition to antifungal agents, but did not show significant clinical improvement. After a positive corneal culture with mold, topical amphotericin B was added to the initial regimen. The mold was identified as Scedosporium apiospermum by macroscopic and microscopic morphologies and the nucleotide sequences of a fungal PCR product showing 99% homology with those of S. apiospermum (EF151349). He recovered with good results at 25 days after corneal epithelial debridement. The early diagnosis of S. apiospermum keratitis is very important for proper treatment. It is recommended that molecular diagnostic methods such as fungal PCR and sequencing be done with conventional cultures whenever a fungal infection is suspected.


Accuracy of high-resolution in vivo micro magnetic resonance imaging for measurements of microstructural and mechanical properties of human distal tibial bone.
Micro magnetic resonance imaging (µMRI) is an in vivo imaging method that permits 3D quantification of cortical and trabecular bone microstructure. µMR images can also be used for building microstructural finite element (µFE) models to assess bone stiffness, which highly correlates with bone's resistance to fractures. In order for µMRI-based microstructural and µFE analyses to become standard clinical tools for assessing bone quality, validation with a current gold standard, namely, high-resolution micro computed tomography (µCT), is required. Microstructural measurements of 25 human cadaveric distal tibias were performed for the registered µMR and µCT images, respectively. Next, whole bone stiffness, trabecular bone stiffness, and elastic moduli of cubic subvolumes of trabecular bone in both µMR and µCT images were determined by voxel-based µFE analysis. The bone volume fraction (BV/TV), trabecular number (Tb.N*), trabecular spacing (Tb.Sp*), cortical thickness (Ct.Th), and structure model index (SMI) based on µMRI showed strong correlations with µCT measurements (r(2) = 0.67 to 0.97), and bone surface-to-volume ratio (BS/BV), connectivity density (Conn.D), and degree of anisotropy (DA) had significant but moderate correlations (r(2) = 0.33 to 0.51). Each of these measurements also contributed to one or many of the µFE-predicted mechanical properties. However, model-independent trabecular thickness (Tb.Th*) based on µMRI had no correlation with the µCT measurement and did not contribute to any mechanical measurement. Furthermore, the whole bone and trabecular bone stiffness based on µMRI were highly correlated with those of µCT images (r(2) = 0.86 and 0.96), suggesting that µMRI-based µFE analyses can directly and accurately quantify whole bone mechanical competence. In contrast, the elastic moduli of the µMRI trabecular bone subvolume had significant but only moderate correlations with their gold standards (r(2) = 0.40 to 0.58). We conclude that most microstructural and mechanical properties of the distal tibia can be derived efficiently from µMR images and can provide additional information regarding bone quality.


Sterol 14alpha-demethylase as a potential target for antitrypanosomal therapy: enzyme inhibition and parasite cell growth.
Sterol 14alpha-demethylases (CYP51) serve as primary targets for antifungal drugs, and specific inhibition of CYP51s in protozoan parasites Trypanosoma brucei (TB) and Trypanosoma cruzi (TC) might provide an effective treatment strategy for human trypanosomiases. Primary inhibitor selection is based initially on the cytochrome P450 spectral response to ligand binding. Ligands that demonstrate strongest binding parameters were examined as inhibitors of reconstituted TB and TC CYP51 activity in vitro. Direct correlation between potency of the compounds as CYP51 inhibitors and their antiparasitic effect in TB and TC cells implies essential requirements for endogenous sterol production in both trypanosomes and suggests a lead structure with a defined region most promising for further modifications. The approach developed here can be used for further large-scale search for new CYP51 inhibitors.


Biology of the Sertoli Cell in the Fetal, Pubertal, and Adult Mammalian Testis.
A healthy man typically produces between 50 × 10(6) and 200 × 10(6) spermatozoa per day by spermatogenesis; in the absence of Sertoli cells in the male gonad, this individual would be infertile. In the adult testis, Sertoli cells are sustentacular cells that support germ cell development by secreting proteins and other important biomolecules that are essential for germ cell survival and maturation, establishing the blood-testis barrier, and facilitating spermatozoa detachment at spermiation. In the fetal testis, on the other hand, pre-Sertoli cells form the testis cords, the future seminiferous tubules. However, the role of pre-Sertoli cells in this process is much less clear than the function of Sertoli cells in the adult testis. Within this framework, we provide an overview of the biology of the fetal, pubertal, and adult Sertoli cell, highlighting relevant cell biology studies that have expanded our understanding of mammalian spermatogenesis.


Dysregulation of the mitogen granulin in human cancer through the miR-15/107 microRNA gene group.
Granulin (GRN) is a potent mitogen and growth factor implicated in many human cancers, but its regulation is poorly understood. Recent findings indicate that GRN is regulated strongly by the microRNA miR-107, which functionally overlaps with miR-15, miR-16, and miR-195 due to a common 5' sequence critical for target specificity. In this study, we queried whether miR-107 and paralogs regulated GRN in human cancers. In cultured cells, anti-argonaute RNA coimmunoprecipitation with downstream microarray analyses indicates that GRN mRNA is directly targeted by numerous miR-15/107 miRNAs. We further tested this association in human tumors. MiR-15 and miR-16 are known to be downregulated in chronic lymphocytic leukemia (CLL). Using pre-existing microarray datasets, we found that GRN expression is higher in CLL relative to nonneoplastic lymphocytes (P < 0.00001). By contrast, other prospective miR-15/miR-16 targets in the dataset (BCL-2 and cyclin D1) were not upregulated in CLL. Unlike in CLL, GRN was not upregulated in chronic myelogenous leukemia (CML) where miR-107 paralogs are not known to be dysregulated. Prior studies have shown that GRN is also upregulated, and miR-107 downregulated, in prostate carcinoma. Our results indicate that multiple members of the miR-107 gene group indeed repress GRN protein levels when transfected into prostate cancer cells. At least a dozen distinct types of cancer have the pattern of increased GRN and decreased miR-107 expression. These findings indicate for the first time that the mitogen and growth factor GRN is dysregulated via the miR-15/107 gene group in multiple human cancers, which may provide a potential common therapeutic target.


Root-specific role for Nicotiana benthamiana RDR6 in the inhibition of Chinese wheat mosaic virus accumulation at higher temperatures.
Some viruses only infect plants at cool temperatures but the molecular mechanism underlying this low-temperature dependence remains unclear. Chinese wheat mosaic virus (CWMV, genus Furovirus) was able to infect wheat and Nicotiana benthamiana plants at 16 but not at 24°C. When CWMV-infected plants were transferred to 24°C for 2 weeks, the newly emerged leaves and roots became virus free. Co-infection with Potato virus Y rescued CWMV accumulation in N. benthamiana plants after a temperature shift to 24°C. In transgenic N. benthamiana plants silenced for the N. benthamiana RNA-dependent RNA polymerase 6 (NbRDR6), CWMV was able to accumulate in roots but not in leaves after a temperature shift to 24°C. Deep sequencing of small RNAs showed that, at 16°C, abundant CWMV small interfering (si)RNAs accumulated in infected N. benthamiana plants. Silencing of NbRDR6 increased the abundance of CWMV siRNAs and the generation of siRNAs from hotspots in the CWMV genome. In contrast, when shifted to 24°C for 1 week, CWMV siRNAs were markedly fewer in roots of NbRDR6-silenced than in roots of wild-type plants but were similar in the leaves of those plants. Our results demonstrate the root-specific role of NbRDR6 in the inhibition of CWMV accumulation and biogenesis of CWMV siRNAs at higher temperatures.


The use of a microagglutination assay for the detection of antibodies to Corynebacterium pseudotuberculosis in naturally infected sheep and goat flocks.
Two goat flocks comprising 326 animals and four sheep flocks comprising 343 animals, all with a previously recognized problem of abscesses due to Corynebacterium pseudotuberculosis, were examined for the presence of abscesses and antibody titers to C. pseudotuberculosis as detected by direct microagglutination assay. In sheep there was a strong positive relationship between age and titer (p less than 0.0001). However, the relationship in goats between age and titer could not be determined due to a strong interaction between flock and age. When the relationship between abscesses and titer was examined, it was found that goats with abscesses had higher titers than those that did not (p less than 0.05), whereas there was no difference in titer between sheep with abscesses and those without (p = 0.5753). The sensitivity of the microagglutination test was poor to good for both species (52.3% for goats and 89.7% for sheep). The specificity of the test was fair to poor (64.9% for goats and 21.7% for sheep). Given a disease prevalence of 13.5% for goats and 8.5% for sheep the predictive value of the positive test was very poor (18.9% for goats and 9.6% for sheep) but the predictive value of the negative test was good to excellent (89.7% for goats and 95.8% for sheep). The poor specificity of the test and therefore the positive predictive value may be due in part to the criterion of classification of presence of disease, i.e. presence of an abscess at the time of sampling.(ABSTRACT TRUNCATED AT 250 WORDS)


Coping with chronic pain among younger, middle-aged, and older adults living with neurological injury and disease.
Objective. This article compares use of pain coping strategies among older, middle-aged, and younger adults living with chronic pain and seeks to determine whether the relationship between pain severity and coping is moderated by age. Method. Participants were 464 adults reporting chronic pain secondary to multiple sclerosis, spinal cord injury, or neuromuscular disease. Participants completed a survey including measures of pain severity and the Chronic Pain Coping Inventory. Results. After controlling for clinical and demographic variables, older adults (older than 60) reported a wider range of frequently used strategies and significantly more frequent engagement in activity pacing, seeking social support, and use of coping self-statements than did younger or middle-aged adults. Moderation analyses suggest that, for younger adults, efforts at coping generally increased with greater pain severity, whereas this relationship did not exist for older adults. Discussion. These data suggest differences in the quantity and quality of pain coping among age groups.


Nasal vaccination with troponin reduces troponin specific T-cell responses and improves heart function in myocardial ischemia-reperfusion injury.
Myocardial ischemia with subsequent reperfusion (MI/R) can lead to significant myocardial damage. Ischemia initiates inflammation at the blood-microvascular endothelial cell interface and contributes significantly to both acute injury and repair of the damaged tissue. We have found that MI/R injury in mice is associated with a cellular immune response to troponin. Myocardial cells exclusively synthesize troponin and release the troponin into the bloodstream following injury. Mucosally administered proteins induce T cells that secrete anti-inflammatory cytokines such as IL-10 and transforming growth factor beta at the anatomical site where the protein localizes. We found that nasal administration of the three subunits of troponin (C, I and T isoforms), given prior to or 1 h following MI/R, decreased infarct size by 40% measured 24 h later. At 1.5 months following MI/R, there was a 50% reduction in infarct size and improvement in cardiac function as measured by echocardiography. Protection was associated with a reduction of cellular immunity to troponin. Immunohistochemistry demonstrated increased IL-10 and reduced IFN-gamma in the area surrounding the ischemic infarct following nasal troponin. Adoptive transfer of CD4+ T cells to mice from nasally troponin-treated mice 1 h after the MI/R decreased infarct size by 72%, whereas CD4+ T cells from IL-10-/- mice or nasally BSA-treated mice had no effect. Our results demonstrate that IL-10-secreting CD4+ T cells induced by nasal troponin reduce injury following MI/R. Modulation of cardiac inflammation by nasal troponin provides a novel treatment to decrease myocardial damage and enhance recovery after myocardial ischemia.


A non-synonymous single-nucleotide polymorphism associated with multiple sclerosis risk affects the EVI5 interactome.
Despite recent progress in the characterization of genetic loci associated with multiple sclerosis (MS) risk, the ubiquitous linkage disequilibrium operating across the genome has stalled efforts to distinguish causative variants from proxy single-nucleotide polymorphisms (SNPs). Here, we have identified through fine mapping and meta-analysis EVI5 as the most plausible disease risk gene within the 1p22.1 locus. We further show that an exonic SNP associated with risk induces changes in superficial hydrophobicity patterns of the coiled-coil domain of EVI5, which, in turns, affects the EVI5 interactome. Immunoprecipitation of wild-type and mutated EVI5 followed by mass spectrometry generated a roster of disease-specific interactors functionally linked to lipid metabolism. Among the exclusive binding partners of the risk variant, we describe the novel interaction with sphingosine 1-phosphate lyase (SGPL1)-a key enzyme for the creation of the sphingosine-1 phosphate gradient, which is relevant to the pathogenic process and therapeutic management of MS.


Mechanisms and targets of deep brain stimulation in movement disorders.
Chronic electrical stimulation of the brain, known as deep brain stimulation (DBS), has become a preferred surgical treatment for medication-refractory movement disorders. Despite its remarkable clinical success, the therapeutic mechanisms of DBS are still not completely understood, limiting opportunities to improve treatment efficacy and simplify selection of stimulation parameters. This review addresses three questions essential to understanding the mechanisms of DBS. 1) How does DBS affect neuronal tissue in the vicinity of the active electrode or electrodes? 2) How do these changes translate into therapeutic benefit on motor symptoms? 3) How do these effects depend on the particular site of stimulation? Early hypotheses proposed that stimulation inhibited neuronal activity at the site of stimulation, mimicking the outcome of ablative surgeries. Recent studies have challenged that view, suggesting that although somatic activity near the DBS electrode may exhibit substantial inhibition or complex modulation patterns, the output from the stimulated nucleus follows the DBS pulse train by direct axonal excitation. The intrinsic activity is thus replaced by high-frequency activity that is time-locked to the stimulus and more regular in pattern. These changes in firing pattern are thought to prevent transmission of pathologic bursting and oscillatory activity, resulting in the reduction of disease symptoms through compensatory processing of sensorimotor information. Although promising, this theory does not entirely explain why DBS improves motor symptoms at different latencies. Understanding these processes on a physiological level will be critically important if we are to reach the full potential of this powerful tool.


Risk factors of vitamin B(12) deficiency in patients receiving metformin.
Identification of risk factors for metformin-related vitamin B(12) deficiency has major potential implications regarding the management of diabetes mellitus. We conducted a nested case-control study from a database in which the source population consisted of subjects who had levels of both serum vitamin B(12) and hemoglobin A(1c) checked in a central laboratory. We identified 155 cases of diabetes mellitus and vitamin B(12) deficiency secondary to metformin treatment. Another 310 controls were selected from the cohort who did not have vitamin B(12) deficiency while taking metformin. A total of 155 patients with metformin-related vitamin B(12) deficiency (mean +/- SD serum vitamin B(12) concentration, 148.6 +/- 40.4 pg/mL [110 +/- 30 pmol/L]) were compared with 310 matched controls (466.1 +/- 330.4 pg/mL [344 +/- 244 pmol/L]). After adjusting for confounders, we found clinically important and statistically significant association of vitamin B(12) deficiency with dose and duration of metformin use. Each 1-g/d metformin dose increment conferred an odds ratio of 2.88 (95% confidence interval, 2.15-3.87) for developing vitamin B(12) deficiency (P<.001). Among those using metformin for 3 years or more, the adjusted odds ratio was 2.39 (95% confidence interval, 1.46-3.91) (P = .001) compared with those receiving metformin for less than 3 years. After exclusion of 113 subjects with borderline vitamin B(12) concentration, dose of metformin remained the strongest independent predictor of vitamin B(12) deficiency. Our results indicate an increased risk of vitamin B(12) deficiency associated with current dose and duration of metformin use despite adjustment for many potential confounders. The risk factors identified have implications for planning screening or prevention strategies in metformin-treated patients.


Policy- and county-level associations with HIV pre-exposure prophylaxis use, the United States, 2018.
HIV pre-exposure prophylaxis (PrEP) is highly efficacious, and yet most individuals indicated for it are not currently using it. To provide guidance for health policymakers, researchers, and community advocates, we developed county-level PrEP use estimates and assessed locality and policy associations. Using data from a national aggregator, we applied a validated crosswalk procedure to generate county-level estimates of PrEP users in 2018. A multilevel Poisson regression explored associations between PrEP use and (1) state policy variables of Medicaid expansion and state Drug Assistance Programs (PrEP-DAPs) and (2) county-level characteristics from the U.S. Census Bureau. Outcomes were PrEP per population (prevalence) and PrEP-to-need ratio (PnR), defined as the ratio of PrEP users per new HIV diagnosis. Higher levels of PrEP prevalence or PnR indicate more PrEP users relative to the total population or estimated need, respectively. Our 2018 county-level data set included a total of 188,546 PrEP users in the United States. Nationally, PrEP prevalence was 70.3/100,000 population and PnR was 4.9. In an adjusted model, counties with a 5% higher proportion of black residents had 5% lower PnR (rate ratio (RR): 0.95, 95% confidence interval (CI): 0.93, 0.96). Similarly, counties with higher concentration of residents uninsured or living in poverty had lower PnR. Relative to states without Medicaid expansion or PrEP-DAPs, states with only one of those programs had 25% higher PrEP prevalence (RR: 1.25, 95% CI: 1.09, 1.45), and states with both programs had 99% higher PrEP prevalence (RR: 1.99, 95% CI: 1.60, 2.48). There was a significant linear trend across the three policy groups, and similar findings for the relation between PnR and the policy groups. In a data set comprising approximately 80% of PrEP users in the United States, we found that Medicaid expansion and PrEP-DAPs were associated with higher PrEP use in states that adopted those policies, after controlling for potential confounders. Future research should identify which components of PrEP support programs have the most success and how to best promote PrEP among groups most impacted by the epidemic. States should support the admirable health decisions of their residents to get on PrEP by implementing policies that facilitate access.


Simple optimization procedure for objective lens correction collar setting.
We present a new method for setting a coverglass correction collar on an objective lens. Axial scans across the interface between the specimen volume and the slide are used together with a quantitative function of merit to determine the optimum setting of the correction collar. The method, which simplifies the adjustment for the user and reduces photobleaching, was implemented within the software environment of a scanning microscope system.


The relationship between individualized care and the practice environment: an international study.
Previous research studies have found that the better the quality of practice environments in hospitals, the better the outcomes for nurses and patients. Practice environment may influence nurses' ability to individualize care but the detailed relationship between individualized care and the professional practice environment has not been investigated widely. Some evidence exists about the association of practice environments with the level of individualization of nursing care, but this evidence is based on single national studies. The aim of this study was to determine whether nurses' views of their professional practice environment associate with their views of the level of care individualization in seven countries. This study had an international, multisite, prospective, cross-sectional, exploratory survey design. The study involved acute orthopedic and trauma surgical inpatient wards (n=91) in acute care hospitals (n=34) in seven countries, Cyprus, Finland, Greece, the State of Kansas, USA, Portugal, Sweden, and Turkey. Nurses (n=1163), registered or licensed practical, working in direct patient care, in orthopedic and trauma inpatient units in acute care hospitals in seven countries participated in the study. Self-administered questionnaires, including two instruments, the Revised Professional Practice Environment and the Individualized Care Scale-Nurse (Individualized Care Scale-Nurse A and B) were used for data collection. Data were analyzed statistically using descriptive statistics, simultaneous multiple regression analysis, and generalized linear model. Two regression models were applied to assess the predictive validity of the Revised Professional Practice Environment on the Individualized Care Scale-Nurse-A and B. The results showed that elements of the professional practice environment were associated with care individualization. Internal work motivation, cultural sensitivity, control over practice, teamwork, and staff relationship with physicians were predictors of support (Individualized Care Scale-A) for and the delivery (Individualized Care Scale-B) of individualized care. The results of this study provide evidence that environment aspect could explain variations in care individualization. These findings support the assertion that individualized care needs to be understood in a broader context than the immediate nurse-patient relationship and that careful development of the care environment may be an effective way to improve care quality and outcomes.


Periodization and physical performance in elite female soccer players.
To investigate the variation in training demands, physical performance, and player well-being across a women's soccer season. Seventeen elite female players wore GPS tracking devices during every training session (N=90) throughout 1 national-league season. Intermittent high-speed-running capacity and 5-, 15-, and 25-m-sprint testing were conducted at the beginning of preseason, end of preseason, midseason, and end of season. In addition, subjective well-being measures were self-reported daily by players over the course of the season. Time over 5 m was lowest at the end of preseason (mean 1.148 s, SE 0.017 s) but then progressively deteriorated to the end of the season (P<.001). Sprint performance over 15 m improved by 2.8% (P=.013) after preseason training, while 25-m-sprint performance peaked at midseason, with a 3.1% (P=.05) improvement from the start of preseason, before declining at the end of season (P=.023). Training demands varied between phases, with total distance and high-speed distance greatest during preseason before decreasing (P<.001) during the early- and late-season phases. Endurance capacity and well-being measures did not change across training phases. Monitoring training demands and subsequent physical performance in elite female soccer players allow coaches to ensure that training periodization goals are being met and related positive training adaptations are being elicited.


How Does Incarcerating Young People Affect Their Adult Health Outcomes?
Despite the widespread epidemic of mass incarceration in the US, relatively little literature exists examining the longitudinal relationship between youth incarceration and adult health outcomes. We sought to quantify the association of youth incarceration with subsequent adult health outcomes. We analyzed data from 14 344 adult participants in the National Longitudinal Study of Adolescent to Adult Health. We used weighted multivariate logistic regressions to investigate the relationship between cumulative incarceration duration (none, <1 month, 1-12 months, and >1 year) before Wave IV (ages 24-34 years) and subsequent adult health outcomes (general health, functional limitations, depressive symptoms, and suicidal thoughts). Models controlled for Wave I (grades 7-12) baseline health, sociodemographics, and covariates associated with incarceration and health. A total of 14.0% of adults reported being incarcerated between Waves I and IV. Of these, 50.3% reported a cumulative incarceration duration of <1 month, 34.8% reported 1 to 12 months, and 15.0% reported >1 year. Compared with no incarceration, incarceration duration of < 1 month predicted subsequent adult depressive symptoms (odds ratio [OR] = 1.41; 95% confidence interval [CI], 1.11-1.80; P = .005). A duration of 1 to 12 months predicted worse subsequent adult general health (OR = 1.48; 95% CI, 1.12-1.96; P = .007). A duration of >1 year predicted subsequent adult functional limitations (OR = 2.92; 95% CI, 1.51-5.64; P = .002), adult depressive symptoms (OR = 4.18; 95% CI, 2.48-7.06; P < .001), and adult suicidal thoughts (OR = 2.34; 95% CI, 1.09-5.01; P = .029). Cumulative incarceration duration during adolescence and early adulthood is independently associated with worse physical and mental health later in adulthood. Potential mechanisms merit exploration.


TFIIH kinase places bivalent marks on the carboxy-terminal domain of RNA polymerase II.
Posttranslational modifications of the carboxy-terminal domain (CTD) of the largest subunit of RNA polymerase II (Pol II) specify a molecular recognition code that is deciphered by proteins involved in RNA biogenesis. The CTD is comprised of a repeating heptapeptide (Y(1)S(2)P(3)T(4)S(5)P(6)S(7)). Recently, phosphorylation of serine 7 was shown to be important for cotranscriptional processing of two snRNAs in mammalian cells. Here we report that Kin28/Cdk7, a subunit of the evolutionarily conserved TFIIH complex, is a Ser7 kinase. The ability of Kin28/Cdk7 to phosphorylate Ser7 is particularly surprising because this kinase functions at promoters of protein-coding genes, rather than being restricted to promoter-distal regions of snRNA genes. Kin28/Cdk7 is also known to phosphorylate Ser5 residues of the CTD at gene promoters. Taken together, our results implicate the TFIIH kinase in placing bivalent Ser5 and Ser7 marks early in gene transcription. These bivalent CTD marks, in concert with cues within nascent transcripts, specify the cotranscriptional engagement of the relevant RNA processing machinery.


The suppressive effect of anti-asialo GM1 antibody on low-dose streptozotocin-induced diabetes in CD-1 mice.
To elucidate the role of natural killer (NK) cells in the pathogenesis of diabetes in streptozotocin-induced diabetes, we examined whether treatment with anti-asialo GM1 antibody prevents the occurrence of diabetes in CD-1 mouse model. Anti-asialo GM1 antibody was injected intraperitoneally 2-3 times a week starting three days before the first streptozotocin injection. In controls, rabbit immunoglobulin or saline was injected instead of anti-asialo GM1 antibody. Three of twelve anti-asialo GM1 antibody-treated mice developed diabetes, however eight of eight (100%) rabbit immunoglobulin injected mice and 20 of 23 saline-injected mice developed diabetes. The incidence of diabetes in the anti-asialo GM1 antibody-injected group was significantly higher than in the two control groups (p less than 0.01, p less than 0.01, respectively). The NK-cell activities of spleen cells from anti-asialo GM1 antibody-treated mice were significantly lower than in control mice. Flow-cytometry analysis demonstrated that anti-asialo GM1 antibody-positive cells had disappeared from spleens of anti-asialo GM1 antibody-injected mice but no suppression of T-lymphocytes could be demonstrated. These results suggest that NK cells play a role in the pathogenesis of streptozotocin-induced diabetes in CD-1 mice.


Crizotinib-Resistant ROS1 Mutations Reveal a Predictive Kinase Inhibitor Sensitivity Model for ROS1- and ALK-Rearranged Lung Cancers.
The identification of molecular mechanisms conferring resistance to tyrosine kinase inhibitor (TKI) is a key step to improve therapeutic results for patients with oncogene addiction. Several alterations leading to EGFR and anaplastic lymphoma kinase (ALK) resistance to TKI therapy have been described in non-small cell lung cancer (NSCLC). Only two mutations in the ROS1 kinase domain responsible for crizotinib resistance have been described in patients thus far. A patient suffering from a metastatic NSCLC harboring an ezrin (EZR)-ROS1 fusion gene developed acquired resistance to the ALK/ROS1 inhibitor crizotinib. Molecular analysis (whole-exome sequencing, CGH) and functional studies were undertaken to elucidate the mechanism of resistance. Based on this case, we took advantage of the structural homology of ROS1 and ALK to build a predictive model for drug sensitivity regarding future ROS1 mutations. Sequencing revealed a dual mutation, S1986Y and S1986F, in the ROS1 kinase domain. Functional in vitro studies demonstrated that ROS1 harboring either the S1986Y or the S1986F mutation, while conferring resistance to crizotinib and ceritinib, was inhibited by lorlatinib (PF-06463922). The patient's clinical response confirmed the potency of lorlatinib against S1986Y/F mutations. The ROS1 S1986Y/F and ALK C1156Y mutations are homologous and displayed similar sensitivity patterns to ALK/ROS1 TKIs. We extended this analogy to build a model predicting TKI efficacy against potential ROS1 mutations. Clinical evidence, in vitro validation, and homology-based prediction provide guidance for treatment decision making for patients with ROS1-rearranged NSCLC who progressed on crizotinib. Clin Cancer Res; 22(24); 5983-91. ©2016 AACR.


The Insertion in Fingers Domain in Human Telomerase Can Mediate Enzyme Processivity and Telomerase Recruitment to Telomeres in a TPP1-Dependent Manner.
In most human cancer cells, cellular immortalization relies on the activation and recruitment of telomerase to telomeres. The telomere-binding protein TPP1 and the TEN domain of the telomerase catalytic subunit TERT regulate telomerase recruitment. TERT contains a unique domain, called the insertion in fingers domain (IFD), located within the conserved reverse transcriptase domain. We report the role of specific hTERT IFD residues in the regulation of telomerase activity and processivity, recruitment to telomeres, and cell survival. One hTERT IFD variant, hTERT-L805A, with reduced activity and processivity showed impaired telomere association, which could be partially rescued by overexpression of TPP1-POT1. Another previously reported hTERT IFD mutant enzyme with similarly low levels of activity and processivity, hTERT-V791Y, displayed defects in telomere binding and was insensitive to TPP1-POT1 overexpression. Our results provide the first evidence that the IFD can mediate enzyme processivity and telomerase recruitment to telomeres in a TPP1-dependent manner. Moreover, unlike hTERT-V791Y, hTERT-V763S, a variant with reduced activity but increased processivity, and hTERT-L805A, could both immortalize limited-life-span cells, but cells expressing these two mutant enzymes displayed growth defects, increased apoptosis, DNA damage at telomeres, and short telomeres. Our results highlight the importance of the IFD in maintaining short telomeres and in cell survival.


Amyloid beta accumulation in axons after traumatic brain injury in humans.
Although plaques composed of amyloid beta (AD) have been found shortly after traumatic brain injury (TBI) in humans, the source for this Abeta has not been identified. In the present study, the authors explored the potential relationship between Abeta accumulation in damaged axons and associated Abeta plaque formation. The authors performed an immunohistochemical analysis of paraffin-embedded sections of brain from 12 patients who died after TBI and from two control patients by using antibodies selective for Abeta peptides, amyloid precursor protein (APP), and neurofilament (NF) proteins. In nine brain-injured patients, extensive colocalizations of Abeta, APP, and NF protein were found in swollen axons. Many of these immunoreactive axonal profiles were present close to Abeta plaques or were surrounded by Abeta staining, which spread out into the tissue. Immunoreactive profiles were not found in the brains of the control patients. The results of this study indicate that damaged axons can serve as a large reservoir of Abeta, which may contribute to Abeta plaque formation after TBI in humans.


Pathological Narcissism: A Study of Burden on Partners and Family.
Pathological narcissism is characterized by impaired interpersonal functioning, but few studies have examined the impact of the disorder on those living in a close relationship. Participants (N = 683; comprising romantic partners [77.8%], mothers [8.5%] or other family members [10%]) in a close relationship with a relative with pathological narcissism completed measures assessing levels of grief, burden, mental health, and coping style. Participants' reported burden was over 1.5 standard deviations above comparison carers of people with mood, neurotic, or psychotic disorders, and higher than carers of people with borderline personality disorder. Similarly, caseness for depression (69% of sample) or anxiety disorders (82%) in the sample was high. Relationship type, subtype expression (vulnerable/grandiose), and coping style were all found to significantly relate to experienced psychopathology. Although limitations exist regarding sample selection that may influence interpretation of results, these findings quantify the significant interpersonal impact of pathological narcissism in this sample.


Hierarchical organization of macaque and cat cortical sensory systems explored with a novel network processor.
Neuroanatomists have described a large number of connections between the various structures of monkey and cat cortical sensory systems. Because of the complexity of the connection data, analysis is required to unravel what principles of organization they imply. To date, analysis of laminar origin and termination connection data to reveal hierarchical relationships between the cortical areas has been the most widely acknowledged approach. We programmed a network processor that searches for optimal hierarchical orderings of cortical areas given known hierarchical constraints and rules for their interpretation. For all cortical systems and all cost functions, the processor found a multitude of equally low-cost hierarchies. Laminar hierarchical constraints that are presently available in the anatomical literature were therefore insufficient to constrain a unique ordering for any of the sensory systems we analysed. Hierarchical orderings of the monkey visual system that have been widely reported, but which were derived by hand, were not among the optimal orderings. All the cortical systems we studied displayed a significant degree of hierarchical organization, and the anatomical constraints from the monkey visual and somato-motor systems were satisfied with very few constraint violations in the optimal hierarchies. The visual and somato-motor systems in that animal were therefore surprisingly strictly hierarchical. Most inconsistencies between the constraints and the hierarchical relationships in the optimal structures for the visual system were related to connections of area FST (fundus of superior temporal sulcus). We found that the hierarchical solutions could be further improved by assuming that FST consists of two areas, which differ in the nature of their projections. Indeed, we found that perfect hierarchical arrangements of the primate visual system, without any violation of anatomical constraints, could be obtained under two reasonable conditions, namely the subdivision of FST into two distinct areas, whose connectivity we predict, and the abolition of at least one of the less reliable rule constraints. Our analyses showed that the future collection of the same type of laminar constraints, or the inclusion of new hierarchical constraints from thalamocortical connections, will not resolve the problem of multiple optimal hierarchical representations for the primate visual system. Further data, however, may help to specify the relative ordering of some more areas. This indeterminacy of the visual hierarchy is in part due to the reported absence of some connections between cortical areas. These absences are consistent with limited cross-talk between differentiated processing streams in the system. Hence, hierarchical representation of the visual system is affected by, and must take into account, other organizational features, such as processing streams.


Shockwave treatment of erectile dysfunction.
Low-intensity extracorporeal shock wave therapy (LI-ESWT) is a novel modality that has recently been developed for treating erectile dysfunction (ED). Unlike other current treatment options for ED, all of which are palliative in nature, LI-ESWT is unique in that it aims to restore the erectile mechanism in order to enable natural or spontaneous erections. Results from basic science experiments have provided evidence that LI-ESWT induces cellular microtrauma, which in turn stimulates the release of angiogenic factors and the subsequent neovascularization of the treated tissue. Extracorporeal shock wave therapy (ESWT) has been clinically investigated and applied in several medical fields with various degrees of success. High-intensity shock wave therapy is used for lithotripsy because of its focused mechanical destructive nature, and medium-intensity shock waves have been shown to have anti-inflammatory properties and are used for treating a wide array of orthopedic conditions, such as non-union fractures, tendonitis, and bursitis. In contrast, LI-ESWT has angiogenetic properties and is therefore used in the management of chronic wounds, peripheral neuropathy, and in cardiac neovascularization. As a result of these characteristics we initiated a series of experiments evaluating the effect of LI-ESWT on the cavernosal tissue of patients with vasculogenic ED. The results of our studies, which also included a double-blind randomized control trial, confirm that LI-ESWT generates a significant clinical improvement of erectile function and a significant improvement in penile hemodynamics without any adverse effects. Although further extensive research is needed, LI-ESWT may create a new standard of care for men with vasculogenic ED.


Charge Transfer at the Qo-Site of the Cytochrome bc1 Complex Leads to Superoxide Production.
The cytochrome bc1 complex is the third protein complex in the electron transport chain of mitochondria or photosynthetic bacteria, and it serves to create an electrochemical gradient across a cellular membrane, which is used to drive ATP synthesis. The purpose of this study is to investigate interactions involving an occasionally trapped oxygen molecule (O2) at the so-called Qo site of the bc1 complex, which is one of the central active sites of the protein complex, where redox reactions are expected to occur. The investigation focuses on revealing the possibility of the oxygen molecule to influence the normal operation of the bc1 complex and acquire an extra electron, thus becoming superoxide, a biologically toxic free radical. The process is modeled by applying quantum chemical calculations to previously performed classical molecular dynamics simulations. Investigations reveal several spontaneous charge transfer modes from amino acid residues and cofactors at the Qo-site to the trapped O2 molecule.


OSCAR is a collagen receptor that costimulates osteoclastogenesis in DAP12-deficient humans and mice.
Osteoclasts are terminally differentiated leukocytes that erode the mineralized bone matrix. Osteoclastogenesis requires costimulatory receptor signaling through adaptors containing immunoreceptor tyrosine-based activation motifs (ITAMs), such as Fc receptor common γ (FcRγ) and DNAX-activating protein of 12 kDa. Identification of these ITAM-containing receptors and their ligands remains a high research priority, since the stimuli for osteoclastogenesis are only partly defined. Osteoclast-associated receptor (OSCAR) was proposed to be a potent FcRγ-associated costimulatory receptor expressed by preosteoclasts in vitro, but OSCAR lacks a cognate ligand and its role in vivo has been unclear. Using samples from mice and patients deficient in various ITAM signaling pathways, we show here that OSCAR costimulates one of the major FcRγ-associated pathways required for osteoclastogenesis in vivo. Furthermore, we found that OSCAR binds to specific motifs within fibrillar collagens in the ECM that become revealed on nonquiescent bone surfaces in which osteoclasts undergo maturation and terminal differentiation in vivo. OSCAR promoted osteoclastogenesis in vivo, and OSCAR binding to its collagen motif led to signaling that increased numbers of osteoclasts in culture. Thus, our results suggest that ITAM-containing receptors can respond to exposed ligands in collagen, leading to the functional differentiation of leukocytes, which provides what we believe to be a new concept for ITAM regulation of cytokine receptors in different tissue microenvironments.


[Prevalence of serotypes and antimicrobial resistance of invasive strains of Streptococcus pneumoniae].
To determine the prevalence of serotypes and antimicrobial susceptibility of invasive strains of Streptococcus pneumoniae and to discuss the implications of these findings for vaccine formulation. Strains of Streptococcus pneumoniae obtained from normally sterile fluids from patients admitted with invasive diseases were isolated and identified at the Hospital de Clínicas, Universidade Federal de Uberlândia, state of Minas Gerais, and forwarded to Instituto Adolfo Lutz, state of São Paulo, for further identification, serotyping and determination of antimicrobial susceptibility. From April 1999 to March 2003, 148 invasive strains of Streptococcus pneumoniae were obtained. The age of patients ranged from 1 day to 88.83 years (mean: 21.33+/-25.82 years; median: 4.42 years). Eighty-four (56.7%) patients were male. The most common diagnoses were pneumonia (91 cases; 61.4%), meningitis (32 cases; 21.6%) and occult bacteremia (15 cases; 10.1%). Strains were isolated mostly from blood (76 occasions; 51.3%), pleural fluid (39 occasions; 26.3%) and cerebrospinal fluid (30 occasions; 20.2%). There were 23 different serotypes, and the most common were 14, 3, 1, 5, 6A, 6B and 18C. Among 30 (20.2%) oxacillin-resistant strains, 23 (15.5%) were confirmed as resistant to penicillin (12.8% intermediate resistance and 2.7% full resistance). Oxacillin-resistant strains were restricted to serotypes 14, 23F, 19A and 6B. Resistance to penicillin varied with age, being more common in children under two years of age (p = 0.0008). We observed decreased sensibility to sulfamethoxazole-trimethoprim (92 isolates; 63.4%), to erythromycin (12 isolates; 8.3%), to clindamycin (12 isolates; 8.7%), to ofloxacin (one strain; 0.8%) and to cefotaxime (three strains; 2%; also resistant to penicillin). All isolates were susceptible to chloramphenicol, rifampin and vancomycin. The decreased susceptibility to penicillin, detected in 15.5% of the strains was predominant in children under two years of age. There were 23 different Streptococcus pneumoniae serotypes. The 23-valent polysaccharide vaccine covers 82.6% of the serotypes and 90.2% of the invasive strains isolated in this population. In addition, 46.7% of the serotypes and 63.6% of the strains isolated from children until five years of age are covered in the currently available 7-valent conjugated vaccine (PN CRM7).


Fuzzy comprehensive evaluation-based disaster risk assessment of desertification in Horqin Sand Land, China.
Desertification is a typical disaster risk event in which human settlements and living environments are destroyed. Desertification Disaster Risk Assessment can control and prevent the occurrence and development of desertification disasters and reduce their adverse influence on human society. This study presents the methodology and procedure for risk assessment and zoning of desertification disasters in Horqin Sand Land. Based on natural disaster risk theory and the desertification disaster formation mechanism, the Desertification Disaster Risk Index (DDRI) combined hazard, exposure, vulnerability and restorability factors and was developed mainly by using multi-source data and the fuzzy comprehensive evaluation method. The results showed that high risk and middle risk areas account for 28% and 23% of the study area, respectively. They are distributed with an "S" type in the study area. Low risk and very low risk areas account for 21% and 10% of the study area, respectively. They are distributed in the west-central and southwestern parts. Very high risk areas account for 18% of the study area and are distributed in the northeastern parts. The results can be used to know the desertification disaster risk level. It has important theoretical and practical significance to prevention and control of desertification in Horqin Sand Land and even in Northern China.


En bloc removal of inflammatory fibrocellular membranes from the iris surface in endophthalmitis.
Dense inflammatory fibrocellular membranes may develop over the iris in an eye with endophalmitis, completely obliterating any view of the posterior segment and hindering vitrectomy. To remove this debris, the edge of the membrane is first lifted from the surface of the iris with a small microvitreoretinal blade inserted through a limbal wound. The membrane is then engaged with an iris hook that is rotated until the debris is tightly wound on the shaft of the instrument. The iris hook is then removed from the anterior segment, delivering the inflammatory debris en bloc. This technique limits possible surgical trauma to the anterior segment and provides an excellent specimen for culture.


Cryoprecipitate for transfusion: which patients receive it and why? A study of patterns of use across three regions in England.
Despite increasing interest in the use of fibrinogen concentrates, cryoprecipitate remains the major source of fibrinogen in England. Understand patterns and indications for use of cryoprecipitate in hospitals from three English regions. Data collection over 3 months from adults, children and neonates receiving cryoprecipitate, including clinical scenario, indications, dose and levels of fibrinogen concentrations pre- and post-transfusion. Four hundred and twenty-three episodes of cryoprecipitate transfusion were analysed from 39 hospitals. Use varied from 0.1 to 4.9 units per 100 red cells transfused. The primary indication was haemorrhage [311 episodes (74%)]. The commonest clinical scenario in all age groups was cardiac surgery, followed by trauma in adults and critical/neonatal care for children. Pre-treatment fibrinogen levels were measured in 322 episodes. In 179 episodes, the level was ≥ 1.0 g L(-1) . Wide variation in practice and dose suggests inconsistent practice and uncertainty in the evidence informing optimal use of cryoprecipitate.


Homodimeric enzymes as drug targets.
Many enzymes and proteins are regulated by their quaternary structure and/or by their association in homo- and/or hetero-oligomer complexes. Thus, these protein-protein interactions can be good targets for blocking or modulating protein function therapeutically. The large number of oligomeric structures in the Protein Data Bank (http://www.rcsb.org/) reflects growing interest in proteins that function as multimeric complexes. In this review, we consider the particular case of homodimeric enzymes as drug targets. There is intense interest in drugs that inhibit dimerization of a functionally obligate homodimeric enzyme. Because amino acid conservation within enzyme interfaces is often low compared to conservation in active sites, it may be easier to achieve drugs that target protein interfaces selectively and specifically. Two main types of dimerization inhibitors have been developed: peptides or peptidomimetics based on sequences involved in protein-protein interactions, and small molecules that act at hot spots in protein-protein interfaces. Examples include inhibitors of HIV protease and HIV integrase. Studying the mechanisms of action and locating the binding sites of such inhibitors requires different techniques for different proteins. For some enzymes, ligand binding is only detectable in vivo or after unfolding of the complexes. Here, we review the structural features of dimeric enzymes and give examples of inhibition through interference in dimer stability. Several techniques for studying these complex phenomena will be presented.


Decreased neutrophil apoptosis in quiescent ANCA-associated systemic vasculitis.
ANCA-Associated Systemic Vasculitis (AASV) is characterized by leukocytoclasis, accumulation of unscavenged apoptotic and necrotic neutrophils in perivascular tissues. Dysregulation of neutrophil cell death may contribute directly to the pathogenesis of AASV. Neutrophils from Healthy Blood Donors (HBD), patients with AASV most in complete remission, Polycythemia Vera (PV), Systemic Lupus Erythematosus (SLE), Rheumatoid Arthritis (RA) and renal transplant recipients (TP) were incubated in vitro, and the rate of spontaneous apoptosis was measured by FACS. Plasma levels of cytokines and sFAS were measured with cytometric bead array and ELISA. Expression of pro/anti-apoptotic factors, transcription factors C/EBP-α, C/EBP-β and PU.1 and inhibitors of survival/JAK2-pathway were measured by real-time-PCR. AASV, PV and RA neutrophils had a significantly lower rate of apoptosis compared to HBD neutrophils (AASV 50 ± 14% vs. HBD 64 ± 11%, p<0.0001). In RA but not in AASV and PV, low apoptosis rate correlated with increased plasma levels of GM-CSF and high mRNA levels of anti-apoptotic factors Bcl-2A1 and Mcl-1. AASV patients had normal levels of G-CSF, GM-CSF and IL-3. Both C/EBP-α, C/EBP-β were significantly higher in neutrophils from AASV patients than HBD. Levels of sFAS were significantly higher in AASV compared to HBD. Neutrophil apoptosis rates in vitro are decreased in AASV, RA and PV but mechanisms seem to differ. Increased mRNA levels of granulopoiesis-associated transcription factors and increased levels of sFAS in plasma were observed in AASV. Additional studies are required to define the mechanisms behind the decreased apoptosis rates, and possible connections with accumulation of dying neutrophils in regions of vascular lesions in AASV patients.


Exploring potential interest in missed period pills in two US states.
Missed period pills (MPP) are uterine evacuation medications used for treatment of delayed menses without prior pregnancy confirmation. This study explores potential interest in missed period pills in two US states. We enrolled people seeking pregnancy test services at nine health centers in two US states between June 2015 and October 2017. Participants completed an anonymous questionnaire containing closed- and open-ended questions about background characteristics, reproductive practices, pregnancy feelings and intentions, abortion attitudes, and MPP interest. We used ordered logistic regression to identify factors associated with MPP interest and inductive content analysis to identify recurring qualitative themes related to MPP interest or disinterest. In all, 678 people completed the survey and 286/678 (42%) indicated interest in missed period pills. Interest was greatest (129/185 or 70%) among those who would be unhappy if pregnant. Variables associated with interest in the multivariate analyses were age ≥ 35, nulliparity, prior abortion and contraceptive use, recent use of emergency contraception, pregnancy feelings and intentions, and abortion attitudes (p < .05). Variables not associated with interest included state of residence, educational attainment, ethnicity, religious affiliation, and frequency of religious attendance. Key reasons for interest were to prevent, avoid or terminate pregnancy; and psychological or emotional benefits, including management of abortion stigma. Reasons for non-interest included concerns about safety or side effects, desire to be pregnant or have a baby, and not wanting to abort or hurt the fetus/baby. If missed period pills were available in the United States, demand might be substantial and wide-ranging across demographic groups. Our findings suggest that some people with missed periods do not desire pregnancy confirmation before taking medications that might disrupt a pregnancy. As a result, provision of missed period pills in the United States would expand reproductive service options and could improve the delivery of patient-centered care.


N-(1,5-Dimethyl-3-oxo-2-phenyl-2,3-di-hydro-1H-pyrazol-4-yl)-2-phenyl-acetamide.
The title compound, C19H19N3O2, crystallizes with two independent mol-ecules (A and B) in the asymmetric unit. In mol-ecule A, the pyrazole ring adopts a slightly disordered half-chair conformation while in B it is planar [r.m.s. deviation = 0.0386 (15) Å]. The dihedral angle between the mean planes of the two phenyl rings is 56.2 (8) in A and 38.2 (3)° in B. The N-phenyl substituent on the pyrazole ring is twisted by 46.5 (2) in A and 58.6 (4)° in B while the extended phenyl ring is twisted by 82.2 (8) in A and 87.5 (9)° in B. The mean plane of the amide group forms an angle of 74.8 (3) in A and 67.7 (1)° in B with respect to the phenyl ring. In addition, the amide group is rotated by 51.4 (1) in A and 53.6 (2)° in B from the the mean plane of the pyrazole ring. In the crystal, the two molecules are linked via N-H⋯O hydrogen bonds, supported by weak C-H⋯O inter-actions, forming dimers enclosing an R 2 (2)(10) ring motif. The dimers are linked via C-H⋯O inter-actions, forming a three-dimensional structure.


Comparison of the incidence and pattern of interstitial lung disease during erlotinib and gefitinib treatment in Japanese Patients with non-small cell lung cancer: the Okayama Lung Cancer Study Group experience.
Data comparing the incidence and pattern of interstitial lung disease (ILD) in non-small cell lung cancer patients receiving treatment with gefitinib versus erlotinib, both of which are epidermal growth factor receptor tyrosine kinase inhibitors, are scarce. We investigated the incidence of ILD in Japanese patients treated with gefitinib or erlotinib. We reviewed the clinical records of 209 patients treated with erlotinib in 2008 (cohort A) and 330 treated with gefitinib between 2000 and 2003 (cohort B). Toxicity within the first month of treatment was investigated. The patients in cohort A had fewer known risk factors for ILD (e.g., poor performance status and prior pulmonary fibrosis). ILD was detected in two patients (1.0%) from cohort A and eight patients (2.4%) from cohort B during the first month of treatment. The events were graded as follows: one patient each in grades 1 and 2 (cohort A), and one, one, and six patients in grades 3, 4, and 5, respectively (cohort B). Multivariate analysis revealed that poor performance status and prior pulmonary fibrosis were significantly correlated with the occurrence of ILD, but the type of epidermal growth factor receptor tyrosine kinase inhibitor administered was not. There was a somewhat lower incidence of ILD with erlotinib therapy than with gefitinib therapy, despite no statistically significant difference. Patient selection based on awareness by Japanese physicians of the risk factors for ILD, rather than the type of agent, may explain the difference in ILD incidence between the two treatments.


Estimating the variation, autocorrelation, and environmental sensitivity of phenotypic selection.
Despite considerable interest in temporal and spatial variation of phenotypic selection, very few methods allow quantifying this variation while correctly accounting for the error variance of each individual estimate. Furthermore, the available methods do not estimate the autocorrelation of phenotypic selection, which is a major determinant of eco-evolutionary dynamics in changing environments. We introduce a new method for measuring variable phenotypic selection using random regression. We rely on model selection to assess the support for stabilizing selection, and for a moving optimum that may include a trend plus (possibly autocorrelated) fluctuations. The environmental sensitivity of selection also can be estimated by including an environmental covariate. After testing our method on extensive simulations, we apply it to breeding time in a great tit population in the Netherlands. Our analysis finds support for an optimum that is well predicted by spring temperature, and occurs about 33 days before a peak in food biomass, consistent with what is known from the biology of this species. We also detect autocorrelated fluctuations in the optimum, beyond those caused by temperature and the food peak. Because our approach directly estimates parameters that appear in theoretical models, it should be particularly useful for predicting eco-evolutionary responses to environmental change.


Severe Listeria monocytogenes infection induces development of monocytes with distinct phenotypic and functional features.
Monocytes perform diverse roles during infection with the facultative intracellular bacterium Listeria monocytogenes. They are essential as bactericidal cells in host defense but can also become Trojan horses transporting bacteria into the brain. To explain these contrasting roles, we characterized bone marrow (BM) monocytes in steady state and generated during lethal and sublethal L. monocytogenes infection. Ly-6C(high)CD11b(+) BM monocytes expressed high amounts of M-CSFR/CD115 in steady state and 72 h following sublethal infection. However, infection with increasing numbers of bacteria resulted in progressive loss of CD115 and strongly decreased CD115-encoding c-fms mRNA expression. Conversely, analysis of regulatory molecules showed de novo expression of the nonsignaling IL-1RII, CD121b, under the same conditions. Ly-6C(high)CD11b(+) monocytes in circulation also acquired a CD115(neg/low)CD121b(high) phenotype during lethal infection. These BM monocytes showed upregulation of suppressor of cytokine signaling 1 and 3 and IL-1R-"associated kinase-M to a greater extent and/or earlier compared with cells from sublethal infection and showed decreased LPS-induced IL-6 production despite similar levels of surface TLR4 expression. BM monocytes from uninfected or sublethally infected mice bound and internalized very few L. monocytogenes in vitro. However, both functions were significantly increased in monocytes developing during lethal infection. Nonetheless, these cells did not produce reactive oxygen intermediates, suggesting an inability to kill L. monocytogenes. Together, these data show that systemic infections with lethal and sublethal amounts of bacteria differentially shape developing BM monocytes. This results in distinct phenotypic and functional properties consistent with being Trojan horses rather than bactericidal effector cells.


Emergency contraception.
Emergency contraceptives are methods that prevent pregnancy when used shortly after unprotected sex. Three different emergency contraceptive methods are safe, simple, and widely available in the United States. These are: (1) ordinary combined oral contraceptives containing ethinyl estradiol and levonorgestrel taken in a higher dose for a short period of time and started within a few days after unprotected intercourse; (2) levonorgestrel-only tablets used similarly; and (3) copper-bearing intrauterine devices inserted within approximately 1 week after unprotected intercourse. Emergency contraceptive use is best known for women who have been raped, but the methods are also appropriate for women who have experienced condom breaks, women who did not use any method because they were not planning on having sex, or women who had unprotected intercourse for any other reason. Unfortunately, few women know about emergency contraceptives, and few clinicians think to inform their patients routinely about the option. A nationwide toll-free hotline (1-888-NOT-2-LATE) and a website (http://not-2-late.com) can help women learn about these options. Sharing "family planning's best-kept secret" widely with women could prevent as many as a million unwanted pregnancies annually in the United States.


Complementary and alternative medicine use decreases adherence to HAART in HIV-positive women.
The use of complementary and alternative medicine (CAM) to treat chronic illnesses, especially HIV, is becoming increasingly widespread. Given this popularity, it is critical to understand how HIV-positive individuals use CAM and, more specifically, whether CAM use impacts their adherence to prescribed antiretroviral regimens (HAART). The present study examined the relationship between CAM use and HAART adherence among HIV+ women. Data were analysed from 366 HIV-positive, mostly African-American women, aged 18-50 years in Alabama and Georgia who were enrolled in an intervention to reduce high-risk sexual behaviour. At enrollment data were collected describing use of CAM and HAART use. Women were classified as CAM users if they reported taking herbal/natural immunity boosters (Chinese herbs, mushrooms, garlic, ginseng or algae) or multivitamins, or reported using religious/psychic health or bodywork to treat HIV. Women were classified as non-adherent if they reported missing any doses of their HAART medication in the 30 days preceding baseline assessment. Logistic regressions models, adjusted for potential confounders, were used to investigate the relationship between CAM use and HAART adherence. Women using CAM (immunity boosters or vitamins), relative to non-CAM users, were 1.69 times more likely to report missing HAART doses in the last 30 days (CI: 1.02-2.80; P=.041) even after adjusting for age, education, race, religion and income. The findings provide preliminary evidence that patients using CAM may be doing so as an alternative to traditional medicine as opposed to complementing prescribed HARRT treatment regimens. The inconsistent use of HAART is problematic given its association with drug resistance. Therefore, health care providers and patients should have explicit dialogues about how to effectively integrate CAM practices into traditional treatment regimens so that the safety and health of HIV-positive patients is not compromised.


Accelerating the carbon cycle: the ethics of enhanced weathering.
Enhanced weathering, in comparison to other geoengineering measures, creates the possibility of a reduced cost, reduced impact way of decreasing atmospheric carbon, with positive knock-on effects such as decreased oceanic acidity. We argue that ethical concerns have a place alongside empirical, political and social factors as we consider how to best respond to the critical challenge that anthropogenic climate change poses. We review these concerns, considering the ethical issues that arise (or would arise) in the large-scale deployment of enhanced weathering. We discuss post-implementation scenarios, failures of collective action, the distribution of risk and externalities and redress for damage. We also discuss issues surrounding 'dirty hands' (taking conventionally immoral action to avoid having to take action that is even worse), whether enhanced weathering research might present a moral hazard, the importance of international governance and the notion that the implementation of large-scale enhanced weathering would reveal problematic hubris. Ethics and scientific research interrelate in complex ways: some ethical considerations caution against research and implementation, while others encourage them. Indeed, the ethical perspective encourages us to think more carefully about how, and what types of, geoengineering should be researched and implemented.


A coordination-based model for transition metal alloy nanoparticles.
We present a simple approach for predicting the relative energies of bimetallic nanoparticles spanning a wide-ranging combinatorial space, using only the identity and nearest-neighbor coordination number of individual metal atoms as independent parameters. By performing straightforward metal atom adsorption calculations on surface slab models, we parameterize expressions for the energy of metal atoms as a function of their coordination number in 21 bimetallic pairings of fcc metals. We rigorously establish the transferability of our model by predicting relative energies of a series of nanoparticles across a large number of morphologies, sizes, atomic compositions, and arrangements. The model is particularly accurate in predicting atomic rearrangements at or near the metal surfaces, which is essential for its potential applications when studying segregation phenomena or dynamic processes in heterogeneous catalysis. By rapidly forecasting site stabilities with atomic specificity across generic structural and compositional features, our model is able to reverse engineer thermodynamically feasible motifs of active sites in bimetallic nanoparticles through robust property ⇔ structure relations.


Variation at the NFATC2 locus increases the risk of thiazolidinedione-induced edema in the Diabetes REduction Assessment with ramipril and rosiglitazone Medication (DREAM) study.
Thiazolidinediones are used to treat type 2 diabetes. Their use has been associated with peripheral edema and congestive heart failure-outcomes that may have a genetic etiology. We genotyped 4,197 participants of the multiethnic DREAM (Diabetes REduction Assessment with ramipril and rosiglitazone Medication) trial with a 50k single nucleotide polymorphisms (SNP) array, which captures ∼2000 cardiovascular, inflammatory, and metabolic genes. We tested 32,088 SNPs for an association with edema among Europeans who received rosiglitazone (n = 965). One SNP, rs6123045, in NFATC2 was significantly associated with edema (odds ratio 1.89 [95% CI 1.47-2.42]; P = 5.32 × 10(-7), corrected P = 0.017). Homozygous individuals had the highest edema rate (hazard ratio 2.89, P = 4.22 × 10(-4)) when compared with individuals homozygous for the protective allele, with heterozygous individuals having an intermediate risk. The interaction between the SNP and rosiglitazone for edema was significant (P = 7.68 × 10(-3)). Six SNPs in NFATC2 were significant in both Europeans and Latin Americans (P < 0.05). Genetic variation at the NFATC2 locus contributes to edema among individuals who receive rosiglitazone.


[Breastfeeding, complimentary feeding practices and childhood malnutrition in the Bolivian Andes].
Northern Potosi is one of the poorest parts of Bolivia with the highest indicators of rural poverty, malnutrition and food insecurity in the Bolivian Andes. The objective of this research was to characterize the levels of malnutrition and describe infant feeding practices in Potosi, Bolivia and use this information to develop an effective, gender sensitive and culturally relevant intervention encouraging good infant feeding practices. Standard methods were used to collect anthropometric data. Weight and height data were collected for 400 children under five years of age from 30 communities. In six of these communities, interviews and focus group discussions were conducted with 33 mothers and other families in addition to household observational data that were collected to describe infant feeding practices. Nearly 20% of children were underweight; stunting was widespread as well. 38% of mothers initiated breastfeeding 12 hours or more after birth. 39% of mothers initiated complementary feeding in the first three months following birth. The type of complementary food given to children was usually inadequate. With this research we could see that nutritional deficiencies often begin when the mother starts breastfeeding and when first introduced complementary feeding. Interventions aimed at improving maternal and child nutrition will require changes in parents' behavior, greater recognition and community support of the importance of child feeding, and the inclusion of strategies to reach young people, involve men, and make high quality nutrition promotion more widely available in the communities.


Monitoring Silent Spillovers Before Emergence: A Pilot Study at the Tick/Human Interface in Thailand.
Emerging zoonoses caused by previously unknown agents are one of the most important challenges for human health because of their inherent inability to be predictable, conversely to emergences caused by previously known agents that could be targeted by routine surveillance programs. Emerging zoonotic infections either originate from increasing contacts between wildlife and human populations, or from the geographical expansion of hematophagous arthropods that act as vectors, this latter being more capable to impact large-scale human populations. While characterizing the viral communities from candidate vectors in high-risk geographical areas is a necessary initial step, the need to identify which viruses are able to spill over and those restricted to their hosts has recently emerged. We hypothesized that currently unknown tick-borne arboviruses could silently circulate in specific biotopes where mammals are highly exposed to tick bites, and implemented a strategy that combined high-throughput sequencing with broad-range serological techniques to both identify novel arboviruses and tick-specific viruses in a ticks/mammals interface in Thailand. The virome of Thai ticks belonging to the Rhipicephalus, Amblyomma, Dermacentor, Hyalomma, and Haemaphysalis genera identified numerous viruses, among which several viruses could be candidates for future emergence as regards to their phylogenetic relatedness with known tick-borne arboviruses. Luciferase immunoprecipitation system targeting external viral proteins of viruses identified among the Orthomyxoviridae, Phenuiviridae, Flaviviridae, Rhabdoviridae, and Chuviridae families was used to screen human and cattle Thai populations highly exposed to tick bites. Although no positive serum was detected for any of the six viruses selected, suggesting that these viruses are not infecting these vertebrates, or at very low prevalence (upper estimate 0.017% and 0.047% in humans and cattle, respectively), the virome of Thai ticks presents an extremely rich viral diversity, among which novel tick-borne arboviruses are probably hidden and could pose a public health concern if they emerge. The strategy developed in this pilot study, starting from the inventory of viral communities of hematophagous arthropods to end by the identification of viruses able (or likely unable) to infect vertebrates, is the first step in the prediction of putative new emergences and could easily be transposed to other reservoirs/vectors/susceptible hosts interfaces.


PEG10 is a c-MYC target gene in cancer cells.
The product of the imprinted gene paternally expressed gene-10 (PEG10) has been reported to support proliferation in hepatocellular carcinomas, but how this gene is regulated has been an open question. We find that MYC knockdown by RNA interference suppresses PEG10 expression in Panc1 pancreatic carcinoma and HepG2 hepatocellular carcinoma cells and that knockdown of PEG10 inhibits the proliferation of Panc1, HepG2, and Hep3B cells. Conversely, PEG10 was up-regulated by inducing c-MYC expression in a B-lymphocyte cell line. Chromatin immunoprecipitation from Panc1 cells showed c-MYC bound to an E-box-containing region in the PEG10 first intron and site-directed mutagenesis showed that the most proximal E-box is essential for promoter activity. In a mouse mammary tumor virus (MMTV)-MYC transgenic mouse model of breast cancer, most but not all of the mammary carcinomas had strongly increased Peg10 mRNA compared with normal mammary gland. By immunohistochemistry, normal human breast and prostate epithelium was negative for the major isoform [reading frame-1 (RF1)] of PEG10 protein, but this cytoplasmic protein was strongly expressed in a subset of breast carcinomas in situ and invasive ductal carcinomas ( approximately 30%) and in a similar percentage of prostate cancers. As in the mouse model, we found positive, but not absolute, correlations between PEG10 and c-MYC in tissue arrays containing 161 human breast cancers (P < 0.002) and 30 prostate cancers (P = 0.014). Immunostaining of human placenta showed PEG10 and c-MYC proteins coexpressed in proliferating cytotrophoblast and coordinately lost in postmitotic syncytiotrophoblast. These findings link cancer genetics and epigenetics by showing that a classic proto-oncogene, MYC, acts directly upstream of a proliferation-positive imprinted gene, PEG10.


Meta-analysis identifies multiple loci associated with kidney function-related traits in east Asian populations.
Chronic kidney disease (CKD), impairment of kidney function, is a serious public health problem, and the assessment of genetic factors influencing kidney function has substantial clinical relevance. Here, we report a meta-analysis of genome-wide association studies for kidney function-related traits, including 71,149 east Asian individuals from 18 studies in 11 population-, hospital- or family-based cohorts, conducted as part of the Asian Genetic Epidemiology Network (AGEN). Our meta-analysis identified 17 loci newly associated with kidney function-related traits, including the concentrations of blood urea nitrogen, uric acid and serum creatinine and estimated glomerular filtration rate based on serum creatinine levels (eGFRcrea) (P < 5.0 × 10(-8)). We further examined these loci with in silico replication in individuals of European ancestry from the KidneyGen, CKDGen and GUGC consortia, including a combined total of ∼110,347 individuals. We identify pleiotropic associations among these loci with kidney function-related traits and risk of CKD. These findings provide new insights into the genetics of kidney function.


Handedness and drinking behaviour.
A number of papers have investigated whether there is an association between handedness and alcohol consumption hypothesizing that alcoholism may be a consequence of atypical cerebral lateralization or a response to the stress involved in being a minority in a right-handed world. Research to date has mostly used small clinical samples, some without a comparison group. This paper exams this issue using a large population-based random sample. A large multi-country data set of nationally representative samples of the non-institutional population aged 50 years and older from 12 European countries was used (N= 27,428). methods: Logistic regression was used to model the frequency with which individuals self-report the frequency of alcohol consumption. A series of models with differing numbers of potential confounders are estimated. The predictors of frequent and infrequent drinking are investigated separately. After controlling for a number of confounders it is shown that left-handers do drink more often. However, this is due to them being less likely to drink rarely (less than once a month) or not at all. The evidence suggests that while there is an association between left-handedness and frequency of alcohol consumption there is no reason to believe that it is associated with excessive alcohol consumption or risky drinking.


Chapter 1. Gene regulatory networks in neural crest development and evolution.
The neural crest is a multipotent migratory embryonic cell population that is present in all vertebrates, but missing from basal chordates. In this chapter, we discuss recent work in amphioxus, ascidians, lamprey, and gnathostomes that reflects the current state of knowledge of the evolutionary origin of this fascinating cell population. We summarize recent evidence for the ongoing diversification of the neural crest in several vertebrate species, with particular reference to studies in nontraditional vertebrate model organisms.


High-performance p-type inorganic-organic hybrid thermoelectric thin films.
The performance of organic-inorganic hybrid thermoelectric thin films can be dramatically enhanced by optimizing energy filtering and carrier transport states at the organic-inorganic interfaces. In this work, p-type "Sb2Te3/CH3NH3I/Sb2Te3" multilayer thin films were firstly fabricated with varied contents of CH3NH3I, and then an annealing process was used in order to form homogeneous organic-inorganic hybrid thin films. The results revealed that the introduced organic component can promote thin film growth and develop a dense nanostructure with improved crystallinity, thus resulting in a significantly increased Seebeck coefficient and a reduced thermal conductivity as a result of the optimized electronic transport characteristics and enhanced effects of phonon scattering. As is expected, the thermoelectric performance of the hybrid-nanocomposite films is enhanced, achieving the maximum ZT value of 1.55 at a temperature of 413 K, which is several times higher than that of the as-fabricated film, thereby suggesting that the proposed strategy can be applied as an efficient method for the preparation of high-performance thermoelectric thin films.


Less depressed or less forthcoming? Self-report of depression symptoms in women preparing for in vitro fertilization.
While depression has been associated with infertility treatments, it is not routinely assessed in women prior to undergoing in vitro fertilization (IVF) treatment. Findings are mixed regarding the degree to which women report depression prior to IVF. The purpose of this study was to (1) examine response profiles in women preparing for IVF and (2) compare responses to those of postpartum, primary care, and general population groups. Female IVF patients (n = 321; 19-45 years) completed the Patient Health Questionnaire-9 (PHQ-9) at their first visit. Clinical and demographic characteristics and incidence of major depressive disorder (MDD) and other depressive disorders (ODD) were examined. Overall score distributions of the IVF group were compared to those of local postpartum patients and published primary care and general populations. Demographic or clinical characteristics did not account for response differences within the IVF group. The IVF group had lower incidences of MDD and ODD than a PHQ-9 normative group. Women in the IVF group reported no depressive symptoms significantly more than postpartum, primary care, and general population groups. Women preparing to undergo IVF report fewer symptoms of depression than multiple comparison groups. Specific quality of life measures may be needed to assess distress in this population.


Synthesis of dimeric ADP-ribose and its structure with human poly(ADP-ribose) glycohydrolase.
Poly(ADP-ribosyl)ation is a common post-translational modification that mediates a wide variety of cellular processes including DNA damage repair, chromatin regulation, transcription, and apoptosis. The difficulty associated with accessing poly(ADP-ribose) (PAR) in a homogeneous form has been an impediment to understanding the interactions of PAR with poly(ADP-ribose) glycohydrolase (PARG) and other binding proteins. Here we describe the chemical synthesis of the ADP-ribose dimer, and we use this compound to obtain the first human PARG substrate-enzyme cocrystal structure. Chemical synthesis of PAR is an attractive alternative to traditional enzymatic synthesis and fractionation, allowing access to products such as dimeric ADP-ribose, which has been detected but never isolated from natural sources. Additionally, we describe the synthesis of an alkynylated dimer and demonstrate that this compound can be used to synthesize PAR probes including biotin and fluorophore-labeled compounds. The fluorescently labeled ADP-ribose dimer was then utilized in a general fluorescence polarization-based PAR-protein binding assay. Finally, we use intermediates of our synthesis to access various PAR fragments, and evaluation of these compounds as substrates for PARG reveals the minimal features for substrate recognition and enzymatic cleavage. Homogeneous PAR oligomers and unnatural variants produced from chemical synthesis will allow for further detailed structural and biochemical studies on the interaction of PAR with its many protein binding partners.


Flow cytometric quantification of glucose-stimulated beta-cell metabolic flux can reveal impaired islet functional potency.
The objective of this study was to develop a multiparametric flow cytometry assay to simultaneously quantify isolated pancreatic islet cell viability, apoptosis, and glucose-induced metabolic flux. INS-1 and rat islet beta-cells were stained with fluorescent probes for cell viability (ToPro3), apoptosis (Annexin V and VADFMK), and intracellular calcium (Ca2+(i)) (Fura Red), stimulated with glucose, and analyzed on a FACS Vantage flow cytometer. Glucose-induced metabolic activity was indicated by changes in Fura Red fluorescence and the autofluorescence of the pyridine [NAD(P)H] and flavin (FAD/FMN) nucleotides. Rat islets cultured under conditions of proinflammatory cytokine-induced oxidative stress were evaluated by flow cytometry and transplantation into diabetic mice. INS-1 and rat islet beta-cell health and metabolic activity were quantified in response to elevated glucose dose and inhibitors of glycolysis and mitochondrial function. Changes in metabolite fluorescence were converted to an area under the curve (AUC) value. Rat islets cultured under oxidative stress conditions showed decreased viability, increased apoptosis, and decreased glucose-induced metabolic activity indicated by reduced AUC for pyridine and flavin nucleotides and Ca2+(i). Reduced metabolite AUC measured by flow cytometry correlated with the inability to reverse diabetes in mice. Single cell flow cytometry can simultaneously quantify both overall islet cell health and beta-cell glucose responsiveness as indicators of functional potency.