After-school interventions to increase physical activity among youth.
Most children and adolescents do not meet the recommended 60 minutes or more of moderate to vigorous physical activity per day. One attractive approach to increasing physical activity in young people is providing activity through structured after-school programmes. This paper provides a review of the scientific literature on the effects of after-school programmes on physical activity in children and adolescents. After-school physical activity interventions provided mixed results; some increased children's physical activity, others did not. Although after-school programmes have the potential to help children and adolescents engage in regular, enjoyable physical activity, the research on these programmes is limited and, in some cases, methodologically weak. Additional, well-controlled studies are needed to identify the components of after-school programmes that promote physical activity and to determine the level of activity that can be attained when children and adolescents participate in these programmes.


Naturalistic driving study of rear seat child occupants: Quantification of head position using a Kinect™ sensor.
Restraint performance is evaluated using anthropomorphic test devices (ATDs) positioned in prescribed, optimal seating positions. Anecdotally, humans-children in particular-assume a variety of positions that may affect restraint performance. Naturalistic driving studies (NDSs), where cameras and other data acquisition systems are placed in a vehicle used by participants during their regular transportation, offer means to collect these data. To date, these studies have used conventional video and analysis methods and, thus, analyses have largely been qualitative. This article describes a recently completed NDS of child occupants in which their position was monitored using a Kinect sensor to quantify their head position throughout normal, everyday driving trips. A study vehicle was instrumented with a data acquisition system to measure vehicle dynamics, a set of video cameras, and a Kinect sensor providing 3D motion capture at 1 Hz of the rear seat occupants. Participant families used the vehicle for all driving trips over 2 weeks. The child occupants' head position was manually identified via custom software from each Kinect color image. The 3D head position was then extracted and its distribution summarized by seat position (left, rear, center) and restraint type (forward-facing child restraint system [FFCRS], booster seat, seat belt). Data from 18 families (37 child occupants) resulted in 582 trips (with children) for analysis. The average age of the child occupants was 45.6 months and 51% were male. Twenty-five child occupants were restrained in FFCRS, 9 in booster seats, and 3 in seat belts. As restraint type moved from more to less restraint (FFCRS to booster seat to seat belt), the range of fore-aft head position increased: 218, 244, and 340 mm on average, respectively. This observation was also true for left-right movement for every seat position. In general, those in the center seat position demonstrated a smaller range of head positions. For the first time in a naturalistic setting, the range of head positions for child occupants was quantified. More variability was observed for those restrained in booster seats and seat belts than for those in FFCRS. The role of activities, in particular interactions with electronic devices, on head position was notable; this will be the subject of further analysis in other components of the broader study. These data can lead to solutions for optimal protection for occupants who assume positions that differ from prescribed, optimal testing positions.


Prevalence of intestinal protozoa infection among school-aged children on Pemba Island, Tanzania, and effect of single-dose albendazole, nitazoxanide and albendazole-nitazoxanide.
Pathogenic intestinal protozoa infections are common in school-aged children in the developing world and they are frequently associated with malabsorption syndromes and gastrointestinal morbidity. Since diagnosis of these parasites is difficult, prevalence data on intestinal protozoa is scarce. We collected two stool samples from school-aged children on Pemba Island, Tanzania, as part of a randomized controlled trial before and 3 weeks after treatment with (i) single-dose albendazole (400 mg); (ii) single-dose nitazoxanide (1,000 mg); (iii) nitazoxanide-albendazole combination (1,000 mg-400 mg), with each drug given separately on two consecutive days; and (iv) placebo. Formalin-fixed stool samples were examined for the presence of intestinal protozoa using an ether-concentration method to determine the prevalence and estimate cure rates (CRs). Almost half (48.7%) of the children were diagnosed with at least one of the (potentially) pathogenic protozoa Giardia intestinalis, Entamoeba histolytica/E. dispar and Blastocystis hominis. Observed CRs were high for all treatment arms, including placebo. Nitazoxanide showed a significant effect compared to placebo against the non-pathogenic protozoon Entamoeba coli. Intestinal protozoa infections might be of substantial health relevance even in settings where they are not considered as a health problem. Examination of a single stool sample with the ether-concentration method lacks sensitivity for the diagnosis of intestinal protozoa, and hence, care is indicated when interpreting prevalence estimates and treatment effects.


Texture classification and segmentation using wavelet frames.
This paper describes a new approach to the characterization of texture properties at multiple scales using the wavelet transform. The analysis uses an overcomplete wavelet decomposition, which yields a description that is translation invariant. It is shown that this representation constitutes a tight frame of l(2) and that it has a fast iterative algorithm. A texture is characterized by a set of channel variances estimated at the output of the corresponding filter bank. Classification experiments with l(2) Brodatz textures indicate that the discrete wavelet frame (DWF) approach is superior to a standard (critically sampled) wavelet transform feature extraction. These results also suggest that this approach should perform better than most traditional single resolution techniques (co-occurrences, local linear transform, and the like). A detailed comparison of the classification performance of various orthogonal and biorthogonal wavelet transforms is also provided. Finally, the DWF feature extraction technique is incorporated into a simple multicomponent texture segmentation algorithm, and some illustrative examples are presented.


Cytotoxicity and in vitro characterization studies of synthesized Jeffamine-cored PAMAM dendrimers.
The objective of this study is to make comprehensive cytotoxicity evaluation and in vitro characterization of Jeffamine-cored polyamidoamine (PAMAM) dendrimers on L929 cell lines for oral drug delivery purposes. Ester-, amine- and carboxylic acid-terminated PAMAMs were investigated for their cytotoxicity on L929 cells at different generations and concentrations. Cationic surface charge caused highest cytotoxicity on L929 cells, while ester-terminated PAMAMs showed generation- and concentration-dependent toxicity. Anionic dendrimers were determined as the lowest cytotoxic group, and highest generation number presented lowest cellular toxicity. Encapsulation studies were performed with anionic PAMAMs at 2.5, 3.5 and 4.5 generations and different concentrations. Increasing generation number provides greater loaded naproxen amounts and larger particle size. Moreover, formulations provide controlled release at simulated terminal ileum conditions. Consequently, Jeffamine-cored carboxylic acid-terminated PAMAMs can be a promising option for oral drug delivery of poorly water-soluble drugs.


Predicting the suicide attempts of lesbian, gay, and bisexual youth.
In this study predictors of serious suicide attempts among lesbian, gay, and bisexual (LGB) youth were examined. Three groups were compared: youth who reported no attempts, youth who reported attempts unrelated to their sexual orientation, and youth whose attempts were considered related to their sexual orientation. About one third of respondents reported at least one suicide attempt; however, only half of the attempts were judged serious based on potential lethality. About half of all attempts were related to youths' sexual orientation. Factors that differentiated youth reporting suicide attempts and those not reporting attempts were greater childhood parental psychological abuse and more childhood gender-atypical behavior. Gay-related suicide attempts were associated with identifiability as LGB, especially by parents. Early openness about sexual orientation, being considered gender atypical in childhood by parents, and parental efforts to discourage gender atypical behavior were associated with gay-related suicide attempts, especially for males. Assessment of past parental psychological abuse, parental reactions to childhood gender atypical behavior, youths' openness about sexual orientation with family members, and lifetime gay-related verbal abuse can assist in the prediction of suicide attempts in this population.


Significant Correlation Between the Infant Gut Microbiome and Rotavirus Vaccine Response in Rural Ghana.
Rotavirus (RV) is the leading cause of diarrhea-related death in children worldwide and 95% of RV-associated deaths occur in Africa and Asia where RV vaccines (RVVs) have lower efficacy. We hypothesize that differences in intestinal microbiome composition correlate with the decreased RVV efficacy observed in poor settings. We conducted a nested, case-control study comparing prevaccination, fecal microbiome compositions between 6-week old, matched RVV responders and nonresponders in rural Ghana. These infants' microbiomes were then compared with 154 age-matched, healthy Dutch infants' microbiomes, assumed to be RVV responders. Fecal microbiome analysis was performed in all groups using the Human Intestinal Tract Chip. We analyzed findings in 78 Ghanaian infants, including 39 RVV responder and nonresponder pairs. The overall microbiome composition was significantly different between RVV responders and nonresponders (FDR, 0.12), and Ghanaian responders were more similar to Dutch infants than nonresponders (P = .002). RVV response correlated with an increased abundance of Streptococcus bovis and a decreased abundance of the Bacteroidetes phylum in comparisons between both Ghanaian RVV responders and nonresponders (FDR, 0.008 vs 0.003) and Dutch infants and Ghanaian nonresponders (FDR, 0.002 vs 0.009). The intestinal microbiome composition correlates significantly with RVV immunogenicity and may contribute to the diminished RVV immunogenicity observed in developing countries.


Otolaryngology Resident Education and the Accreditation Council for Graduate Medical Education Core Competencies: A Systematic Review.
To date, there have been no reports in the current literature regarding the use of the Accreditation Council for Graduate Medical Education (ACGME) core competencies in otolaryngology residency training. An evaluation may help educators address these core competencies in the training curriculum. To examine the quantity and nature of otolaryngology residency training literature through a systematic review and to evaluate whether this literature aligns with the 6 core competencies. A medical librarian assisted in a search of all indexed years of the PubMed, Embase, Education Resources Information Center (via EBSCOhost), Cochrane Library (Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, and Cochrane Methodology Register), Thomson Reuters Web of Science (Science Citation Index Expanded, Social Sciences Citation Index Expanded, Conference Proceedings Citation Index-Science, and Conference Proceedings Citation Index-Social Science and Humanities), Elsevier Scopus, and ClinicalTrials.gov databases to identify relevant English-language studies. Included studies contained original human data and focused on otolaryngology resident education. Data regarding study design, setting, and ACGME core competencies addressed were extracted from each article. Initial searches were performed on May 20, 2015, and updated on October 4, 2016. In this systematic review of 104 unique studies, interpersonal communication skills were reported 15 times; medical knowledge, 48 times; patient care, 44 times; practice-based learning and improvement, 31 times; professionalism, 15 times; and systems-based practices, 10 times. Multiple studies addressed more than 1 core competency at once, and 6 addressed all 6 core competencies. Increased emphasis on nonclinical core competencies is needed, including professionalism, interpersonal and communication skills, and systems-based practices in the otolaryngology residency training curriculum. A formal curriculum addressing nonclinical core competencies should be integrated into otolaryngology residency training.


Mitochondrial control of apoptosis: an overview.
The mitochondrial permeability transition (PT) pore, also called the mitochondrial megachannel, is a multiprotein complex formed at the contact site between the mitochondrial inner and outer membranes, exactly the same location at which Bax, Bcl-2 and Bcl-XL are particularly abundant. The PT pore participates in the regulation of matrix Ca2+, pH, transmembrane potential and volume, and functions as a Ca(2+)-, voltage-, pH- and redox-gated channel with several levels of conductance and little, if any, ion selectivity. We have obtained three independent lines of evidence implicating the mitochondrial PT pore in apoptosis. First, in intact cells, apoptosis is accompanied by an early dissipation of the mitochondrial transmembrane potential, delta psi m. In several models of apoptosis, specific agents inhibiting the mitochondrial PT pore abolish this dissipation of the delta psi m and simultaneously prevent activation of downstream caspases and endonucleases, indicating that PT pore opening can be a critical event of the apoptotic process. Secondly, mitochondria are rate-limiting for caspase and nuclease activation in several cell-free systems of apoptosis. Isolated mitochondria release apoptogenic factors capable of activating pro-caspases or endonucleases upon opening of the mitochondrial megachannel in vitro. Thirdly, opening of the purified PT pore complex reconstituted into liposomes is inhibited by recombinant Bcl-2 or Bcl-XL, two apoptosis-inhibitory proteins that also prevent PT pore opening in cells and isolated mitochondria. Altogether, our results suggest that PT pore opening is sufficient and (mostly) necessary for triggering apoptosis. The implications of these findings are examined in the light of pharmacological interventions in apoptosis.


Effects of anesthetic agents on autoregulation of renal hemodynamics in the rat and dog.
Controversy has existed over apparent dissimilarities in the autoregulatory capacities of the rat and dog. A protocol was designed to evaluate both the effects of the anesthetic agents. Nembutal (used most commonly in dogs) and Inactin (most frequently employed in rats) and the species peculiarities of these two mammals on autoregulation of renal blood flow (RBF) and glomerular filtration rate (GFR). With Nembutal autoregulation of RBF was present in both experimental animals. Inactin impaired RBF autoregulation similarly in both species. With impaired RBF autoregulation similarly in both species. With either anesthetic GFR was autoregulated well in both rat and dog. Comparison of the two species revealed a greater RBF per gram kidney weight and a higher renal perfusion pressure (RPP) at which autoregulation of both hemodynamic parameters was lost in the rat. It is concluded from these studies that 1) the frequent use of Inactin in the rat in large part accounts for the observed lack of autoregulation of RBF in this animal and 2) renal hemodynamic responses are qualitatively similar in rat and dog when the same anesthetic agents are used.


Postoperative care for Samter's triad patients undergoing endoscopic sinus surgery: a double-blinded, randomized controlled trial.
Evidence is lacking to guide the postoperative management of Samter's triad patients with chronic rhinosinusitis with polyposis (CRSwP) undergoing endoscopic sinus surgery (ESS). The purpose of this study was to compare three different standardized medication regimens prescribed to these patients after ESS. Three-arm, randomized, double-blinded, controlled trial. Patients with Samter's triad undergoing ESS were postoperatively randomized into three medication regimens, those being saline irrigation alone (control group A), saline irrigation plus separate budesonide nasal spray (group B), and saline irrigation mixed with budesonide nasal spray (group C). Outcome measures were Sino-Nasal Outcome Test scores, Lund-Mackay computed tomography scores, and Lund-Kennedy endoscopic scores taken at preoperative baseline, and then at 6 months and 1 year postoperatively. Side effect profiles were also measured (adrenocorticotropic hormone blood level ranges and intraocular pressure at the same interval points). Analysis of variance and χ(2) analyses were conducted using a Bonferroni correction method and routine descriptive statistics. Inter- and intragroup comparisons were made. Sixty subjects were recruited. All groups were equivalent at baseline in all outcomes. All intragroup analyses showed statistically and clinically significant improvement in disease status as compared to baseline (P < .0167), with a sustained but lessened improvement at 1 year. However, no statistically or clinically significant differences were observed between groups at any time point (P > .05). There was no treatment effect noted. In this study, nasal steroids did not confer any additional benefit over saline alone as post-ESS care for the Samter's triad CRSwP patient population.


Attachment and ingestion of bacteria by trophozoites of Entamoeba histolytica.
Entamoeba histolytica trophozoites were found to be very selective in their interactions with bacteria. Two principal mechanisms were shown to be responsible for these interactions. Certain bacteria, such as a number of Escherichia coli and Serratia marcescens strains which are known to contain mannose-binding components on their cell surface, bound to mannose receptors on the amoeba membrane. This attachment was markedly inhibited by alpha-methylmannoside (0.5%), especially when the incubations were done at low temperature (5 degrees C). Other bacterial species, such as Shigella flexneri and Staphylococcus aureus, which do not possess a mannose-binding capacity, attached to the amoebae, but only with the aid of concanavalin A or after opsonization of the bacteria with immune serum. In both types of attachment, between 40 and 100 bacteria bound per amoeba, and considerable ingestion of bacteria into amoeba vacuoles was observed at 37 degrees C. The attachment of opsonized bacteria to the amoebae does not appear to be mediated by Fc receptors since Fab' dimers obtained after pepsin digestion of immunoglobulin were capable of mediating adherence. Furthermore, preincubation of the amoebae with aggregated human immunoglobulin G or with heat-inactivated immune serum and EDTA did not inhibit the attachment of opsonized bacteria. The attachment of opsonized bacteria was markedly inhibited by N-acetylglucosamine-containing glycoconjugates, such as peptidoglycan and chitin oligosaccharides, as well as by N-acetylgalactosamine. These results indicate that amoebae can attach and ingest bacteria either by using their membrane-associated carbohydrate-binding protein or by having their mannose-containing cell surface components serve as receptors.


COVID-19 vaccine hesitancy among rural Oklahomans.
COVID-19 vaccination is widely recommended as a prevention strategy; however, vaccine uptake is disproportionately lower among rural Americans compared to their urban counterparts. Development of public health activities to address the rural-urban vaccine gap requires an understanding of determinants of vaccine hesitation. The present study explores perceptions of and barriers to COVID-19 vaccination among rural Oklahomans. Between March and May 2021, 222 residents, unvaccinated for COVID-19, within rural Oklahoma counties completed a cross-sectional, online questionnaire to qualitatively assess perceptions, benefits, and concerns regarding getting vaccinated for COVID-19. Approximately two-fifths of rural respondents in the present study were hesitant to get vaccinated, even when a vaccine was made available to them. Major factors included limited knowledge and understanding about the vaccine, including potential side-effects and long-term complications, as well as skepticism surrounding COVID-19 vaccine development and efficacy. Among the potential perceived benefits of vaccination were protecting the health of vulnerable individuals and the ability to return to normal day-to-day activities. Increases in COVID-19 cases and deaths in rural areas are expected to continue as new variants are introduced within communities. The present findings highlight the need for the development of culturally tailored vaccine information, to be disseminated by local leaders within rural communities.


In vitro susceptibility of dermatophytes to oral antifungal drugs and amphotericin B in Uttar Pradesh, India.
Dermatophytosis is a major public health problem in our country. Although resistance to conventional oral and topical antifungal agents is being increasingly encountered, the sensitivity pattern of dermatophytes has not been systematically analysed. We aimed to determine the sensitivity pattern of dermatophyte isolates to amphotericin B and six oral antifungal drugs. Patients with dermatophytosis attending the outpatient department of dermatology were enrolled in the study. Samples were collected for mycological examination and in vitro antifungal sensitivity testing was done by broth microdilution as per the Clinical and Laboratory Standard Institute M38-A standards. A total of 804 patients were enrolled. Specimens from 185 patients (23%) were both KOH and culture positive, and 44 of these isolates (41 Trichophyton mentagrophytes and 3 Trichophyton rubrum) were subjected to sensitivity testing. Minimum inhibitory concentrations (MIC) of itraconazole, ketoconazole, voriconazole and amphotericin B were comparable. The median MIC to fluconazole was higher than the other tested drugs. Dermatophytes were most susceptible to ketoconazole and voriconazole, followed by itraconazole, amphotericin B, fluconazole and griseofulvin. A high incidence of resistance was found to terbinafine and the difference was statistically significant in comparison to fluconazole, itraconazole, voriconazole, ketoconazole (P = 0.001) and griseofulvin (P = 0.003). The strains were more sensitive to amphotericin B as compared to griseofulvin (P = 0.02) and terbinafine (P < 0.001). This was a hospital-based study and may not reflect the true pattern in the community. Only a few of the isolates were selected for study. The clinical response of patients, whose isolates were studied for in vitro sensitivity of the antifungals, was not studied. The sensitivity pattern of dermatophytes to various antifungals including amphotericin B, ketoconazole, voriconazole and itraconazole were determined. The studied isolates were least susceptible to terbinafine.


When reciprocity fails: effort-reward imbalance in relation to coronary heart disease and health functioning within the Whitehall II study.
A deleterious psychosocial work environment, as defined by high efforts expended in relation to few rewards reaped, is hypothesised to increase the risk of future poor health outcomes. To test this hypothesis within a cohort of London based civil servants. Effort-reward imbalance (ERI) was measured among 6895 male and 3413 female civil servants aged 35-55 during the first phase of the Whitehall II study (1985-88). Participants were followed until the end of phase 5 (1997-2000), with a mean length of follow up of 11 years. Baseline ERI was used to predict incident validated coronary heart disease (CHD) events during follow up and poor mental and physical functioning at phase 5. A high ratio of efforts in relation to rewards was related to an increased incidence of all CHD (hazard ratio (HR) = 1.36, 95% CI 1.12 to 1.65) and fatal CHD/non-fatal myocardial infarction (HR = 1.28, 95% CI 0.89 to 1.84) during follow up, as well as poor physical (odds ratio (OR) = 1.47, 95% CI 1.24 to 1.74) and mental (OR = 2.24, 95% CI 1.89 to 2.65) functioning at phase 5, net of employment grade. A one item measure of high intrinsic effort also significantly increased the risk of these health outcomes, net of grade. ERI may be particularly deleterious with respect to CHD risk among those with low social support at work or in the lowest employment grades. Within the Whitehall II study, a ratio of high efforts to rewards predicted higher risk of CHD and poor physical and mental health functioning during follow up. Although the increased risk associated with ERI was relatively small, as ERI is common it could be of considerable public health importance.


Efficacy of Arabica Versus Robusta Coffee in Improving Weight, Insulin Resistance, and Liver Steatosis in a Rat Model of Type-2 Diabetes.
The effects of chronic coffee exposure in models of type 2 diabetes mellitus (T2D) models is scarcely studied, and the efficacy of the main coffee species has never been compared. We tested the hypothesis that long-term consumption of arabica and robusta coffee may differentially delay and affect T2D development in Zucker diabetic fatty rats. Three study groups received either chow mixed with arabica or robusta instant coffee (1.8% w/w) or unsupplemented chow food for 10 weeks. Both coffee species reduced liver triglyceride content and area under the curve of fasting and postprandial insulin. At study end, plasma adiponectin, total cholesterol and high density lipoprotein levels were higher in the robust group compared with both arabica and control groups. The liver gene expression of Glucose-6-phosphatase, catalytic subunit (G6pc) and Mechanistic target of rapamycin (mTOR) in robusta and Cpt1a in both coffee groups was downregulated. In conclusion, long-term consumption of both coffee species reduced weight gain and liver steatosis and improved insulin sensitivity in a rat model of T2D. Robusta coffee was seemingly superior to arabica coffee with respect to effects on lipid profile, adiponectin level and hepatic gene expression.


Stabilization of the Trigonal Langasite Structure in Ca3Ga2-2xZnxGe4+xO14 (0 ≤ x ≤ 1) with Partial Ordering of Three Isoelectronic Cations Characterized by a Multitechnique Approach.
Crystallization of oxide glasses rich in Zn2+, Ga3+, and Ge4+ is of interest for the synthesis of new transparent ceramics. In this context, we report the identification and detailed structural characterization of a new solid solution Ca3Ga2-2xZnxGe4+xO14 (0 ≤ x ≤ 1). These compounds adopt the trigonal langasite structure type, offering three possible crystallographic sites for the coordination of isoelectronic Zn2+, Ga3+, and Ge4+. We used neutron diffraction to determine distributions of Ga3+/Ge4+ and Zn2+/Ge4+ in the simpler end members Ca3Ga2Ge4O14 and Ca3ZnGe5O14, while for the complex intermediate member Ca3GaZn0.5Ge4.5O14, we used an original approach combining quantitative 2D analysis of atomic-resolution STEM-EDS maps with neutron diffraction. This revealed that, across the solid solution, the tetrahedral D sites remain fully occupied by Ge4+, while Zn2+, Ga3+, and the remaining Ge4+ are shared between octahedral B- and tetrahedral C sites in proportions that depend upon their relative ionic radii. The adoption of the trigonal langasite structure by glass-crystallized Ca3ZnGe5O14, a compound that was previously observed only in a distorted monoclinic langasite polymorph, is attributed to substantial disorder between Zn2+ and Ge4+ over the B and C sites. The quantitative 2D refinement of atomic-resolution STEM-EDS maps is applicable to a wide range of materials where multiple cations with poor scattering contrast are distributed over different crystallographic sites in a crystal structure.


Cardiac involvement in non-human primates infected with the Lyme disease spirochete Borrelia burgdorferi.
To investigate cardiac involvement in the non-human primate (NHP) model of Lyme disease, we inoculated 39 adult Macaca mulatta with Borrelia burgdorferi sensu stricto strains N40 (BbN40) by needle (N=22, 14 immunocompetent (IC), seven permanently immunosuppressed (IS), and four transiently immunosuppressed (TISP)) or by tick-bite (N=4, all TISP) or strain 297 (Bb297) by needle (N=2 IS), or with B. garinii strains Pbi (N=4, 2 TISP and 2 IS), 793 (N=2, TISP) or Pli (N=2, TISP). Five uninfected NHPs were used as controls. Infection and inflammation was studied in the hearts and the aorta removed at necropsy 2-32 months after inoculation by (1) H&E and trichrome-staining; (2) immunohistochemistry and digital image analysis; (3) Western blot densitometry; and (4) TaqMan RT-PCR. All NHPs inoculated with BbN40 became infected and showed carditis at necropsy. The predominant cells were T cells, plasma cells, and macrophages. There was increased IgG and IgM in the heart independent of immunosuppression. The B-cell chemokine BLC was significantly increased in IS-NHPs. There was increased deposition of the complement membrane attack complex (MAC) in TISP and IS-NHPs. The spirochetal load was very high in all BbN40-inoculated IS-NHPs but minimal if any in IC or TISP NHPs. Double-immunostaining revealed that many spirochetes in the heart of BbN40-IS NHPs had MAC on their membranes. We conclude that carditis in NHPs infected with B. burgdorferi is frequent and can persist for years but is mild unless they are immunosupressed.


Identification of Polo-like kinases as potential novel drug targets for influenza A virus.
In recent years genome-wide RNAi screens have revealed hundreds of cellular factors required for influenza virus infections in human cells. The long-term goal is to establish some of them as drug targets for the development of the next generation of antivirals against influenza. We found that several members of the polo-like kinases (PLK), a family of serine/threonine kinases with well-known roles in cell cycle regulation, were identified as hits in four different RNAi screens and we therefore studied their potential as drug target for influenza. We show that knockdown of PLK1, PLK3, and PLK4, as well as inhibition of PLK kinase activity by four different compounds, leads to reduced influenza virus replication, and we map the requirement of PLK activity to early stages of the viral replication cycle. We also tested the impact of the PLK inhibitor BI2536 on influenza virus replication in a human lung tissue culture model and observed strong inhibition of virus replication with no measurable toxicity. This study establishes the PLKs as potential drug targets for influenza and contributes to a more detailed understanding of the intricate interactions between influenza viruses and their host cells.


Radical-Pair-Based Magnetoreception Amplified by Radical Scavenging: Resilience to Spin Relaxation.
Birds and several other species are equipped with the remarkable ability to sense the geomagnetic field for the purpose of navigation and orientation. The primary detection mechanism of this compass sense is uncertain but appears to originate from a truly quantum process involving spin-correlated radical pairs. In order to elicit sensitivity to weak magnetic fields, such as the Earth's magnetic field, the underlying spin dynamics must be protected from fast decoherence. In this work, we elucidate the effects of spin relaxation on a recently suggested reaction scheme involving three radicals, instead of a radical pair, doublet-quartet interconversion under magnetic interactions, and a spin-selective scavenging reaction. We show that, besides giving rise to a vastly enhanced reaction anisotropy, this extended reaction scheme is more resilient to spin relaxation than the conventional radical pair mechanism. Surprisingly, the anisotropic magnetic field effect can be enhanced by fast spin relaxation in one of the radicals of the primary pair. We discuss this finding in the context of magnetoreception. Radical scavenging can protect the spin system against fast spin relaxation in one of the radicals, thereby providing a credible model to the involvement of fast relaxing radical pairs, such as FADH•/O2•-, in radical-pair based magnetoreception. This finding will help explain behavioral observations that seem incompatible with the previously proposed flavin semiquinone/tryptophanyl radical pair.


The RNaseIII enzyme Dicer is required for morphogenesis but not patterning of the vertebrate limb.
The RNaseIII-containing enzyme Dicer is believed to be required for the processing of most, if not all, microRNAs (miRNAs) and for processing long dsRNA into small interfering RNAs. Because the complete loss of Dicer in both zebrafish and mice results in early embryonic lethality, it has been impossible to determine what role, if any, Dicer has in patterning later tissues in the developing vertebrate embryo. To bypass the early requirement of Dicer in development, we have created a conditional allele of this gene in mice. Using transgenes to drive Cre expression in discrete regions of the limb mesoderm, we find that removal of Dicer results in the loss of processed miRNAs. Phenotypically, developmental delays, in part due to massive cell death as well as disregulation of specific gene expression, lead to the formation of a much smaller limb. Thus, Dicer is required for the formation of normal mouse limbs. Strikingly, however, we did not detect defects in basic patterning or in tissue-specific differentiation of Dicer-deficient limb buds.


A needle micro-osmometer for determination of glycosaminoglycan concentration in excised nucleus pulposus tissue.
Aggrecan is one of the major macromolecular components of the intervertebral disc (IVD) and its loss is an early sign of degeneration. Restoration of aggrecan, and hence of biomechanical properties, is a major objective of biological therapies. At present, assessment of aggrecan concentration via its glycosaminoglycan (GAG) content is accomplished using biochemical and histological methods which require sacrifice of tissue. A minimally invasive method for assessing GAG, and hence aggrecan, which can avoid destruction of tissue, would be of benefit. We have developed a needle micro-osmometer that is capable of measuring flux of saline into excised human nucleus pulposus (NP) tissue. Using the isotropic osmotic stress technique to assess the swelling pressure of the excised NP tissue and assuming negligible collagen tensile stress, we were able to relate the flux to the tissue fixed charge density (FCD). GAG concentration is evaluated from its FCD via the radioactive tracer technique. Samples representing different ages (28-59 years) and degeneration grades (1-4) were analyzed. The flux is controlled by both the osmotic pressure difference across the probe's semi-permeable membrane and by the tissue permeability. A linear correlation was found between flux and the tissue FCD. The equation describing the linear fit is FCD/(total tissue hydration) = 1.97 × 10(-4) + 8283 × flux (R = 0.836, p < 10(-4)). Thus, by measuring saline flux, the concentration of GAG can be determined. Micro-osmometry provides a reliable and minimally invasive tool for assessing GAG content in excised NP tissue. This method may be usefully applied in tissue engineering applications. It may also be useful for in vivo measurements if the question of the degenerative effect of needle puncture can be overcome.


The impact of parenting styles on children developmental outcome: The role of academic self-concept as a mediator.
Although the importance of parenting styles directly influencing child development is well established, fewer studied have examined whether parenting styles also affect children's behavioural problems indirectly, mediated through children's academic self-concept (ASC). We examined direct and shared effects of parenting styles on behavioural problems of 199 Kurdish primary school children with a mean age of 11 years 7 months (range 11 years 5 months to 12 years 3 months). Questionnaires measured parenting styles (child version of Alabama Parenting Questionnaire), assessed children's ASC (Myself-As-Learner Scale) and identified children's behavioural problems with the Strengths and Difficulties Questionnaire (SDQ). PROCESS analysis was used to perform the mediation analysis. The results revealed that positive and negative parenting composites are indirectly related to children's internalising behaviour problems. In addition, ASC partially mediated the relationship between the negative parenting composite and prosocial behaviour. However, the mediation analysis did not show the expected indirect effect of parenting styles on externalising problems as being mediated via ASC. Hence, we argue that the ASC serves as a significant mediator in the relationship between parenting styles with prosocial behaviour and internalising problems.


MSH-2 and MLH-1 protein expression in Muir Torre syndrome-related and sporadic sebaceous neoplasms.
Muir-Torre Syndrome (MTS) is a rare autosomal-dominant disorder characterized by the predisposition to both sebaceous neoplasm and internal malignancies. MTS-associated sebaceous neoplasms reveal mutations in DNA mismatch repair (MMR) genes and microsatellite instability. A significant part of MTS patients represents a phenotypic variant, the hereditary nonpolyposis colorectal cancer (HNPCC). A strong correlation between microsatellite instability and immunostaining has been demonstrated. The early recognition of sebaceous neoplasm as part of MTS, and their differentiation from sporadic sebaceous neoplasm may have an important application in a clinical setting. The absence of MLH-1 or MSH-2 expression by immunostaining identifies tumors with mismatch repair deficiency. Our aim is to determine whether an immunohistochemical approach, targeting DNA repair proteins MSH-2 and MLH-1 in MTS-related sebaceous neoplasm and their sporadic counterparts, can be used for their identification. We examined 15 sebaceous neoplasms (including 6 internal malignancy- associated sebaceous neoplasms and 8 sporadic sebaceous neoplasms) from 11 patients for the expression of MSH-2 and MLH-1 by immunohistochemistry. Four of 5 internal malignancy-associated sebaceous neoplasms showed loss of expression of MSH-2 or MLH-1. Correlation of the immunostaining pattern of the sebaceous neoplasms and the patients' positive history of colon carcinoma was 80%. Seven of 8 sporadic sebaceous neoplasms showed a positive expression of MSH-2 and MLH-1. The prevalence for loss of expression of MMR proteins in sebaceous neoplasms was 38.5%. MMR immunostaining had 87.5% specificity and 80% sensitivity. This study is limited by a small sample size, and by bias selection due to the use of non nationwide data-base as the resource of cases. Our findings demonstrate that immunohistochemical testing for internal malignancy-associated sebaceous neoplasms is a practical approach to confirm a suspected inherited MMR gene defect, and an accurate method to distinguish between sporadic and MTS-associated sebaceous lesions.


Follow-up study of patients treated by x-ray epilation for tinea capitis. Estimation of the dose to the thyroid and pituitary glands and other structures of the head and neck.
This study is a further investigation of radiation dose to various head structures in the children given X-ray therapy for tinea capitis (ringworm of the scalp). In this work, estimates of the dose to the thyroid and pituitary gland were obtained with lithium fluoride thermoluminescent dosemeters using a child's head phantom. Doses were also measured for the parotid gland and several skin sites where skin tumours developed in the irradiated cases. In a previous study, brain and scalp doses of 140 and 500-800 rad had been estimated for the treated group using this same head phantom. In this work dosemeters were also placed in the same brain locations so that comparisons could be obtained between the two studies. The thyroid dose was estimated to be 6 +/- 2 rad and the pituitary dose was 49 +/- 6 rad for the conventional tinea capitis treatment. The dose to the parotid gland was 39 rad and the dose to skin sites on the face and neck where tumours occurred ranged from 20 to 40 rad. The data for the thyroid adenoma response from this and other studies involving irradiation of children suggests a linear dose-response relationship within the first 30-40 years after exposure with a risk of about 0-04% per rad.


Scanning electron microscopy analysis of RinsEndo system and conventional irrigation for debris removal.
A new device for irrigation, which presents hydrodynamic activation based on the pressure-suction technology, has recently been introduced to the market: the RinsEndo system. This study compared the efficacy of the RinsEndo system and conventional (manual-dynamic) irrigation in the removal of debris from the root canal walls, using scanning electron microscopy (SEM). Twenty mandibular premolars with completely formed roots were selected and randomly divided into group 1 (irrigation with the RinsEndo system) and group 2 (conventional irrigation). The canals were irrigated with 1 mL of saline at each change of instrument. Instrumentation started with a #15 K file and continued up to a #40 K file, which was standardized as the working length instrument. Then, the teeth were sectioned in buccolingual direction and the halves were sputter-coated with gold and examined by SEM. The apical, middle and cervical root canal thirds were evaluated, and the results were analyzed statistically by the Mann-Whitney test for comparison between methods, Kruskal-Wallis test for comparison among thirds, and Miller test for individual comparisons. A significance level of 5% was set for all analyses. The results did not show significant differences (p>0.05) between methods at each third and among thirds for each technique analyzed individually. In conclusion, there was no difference in the cleaning ability of the RinsEndo system and conventional irrigation.


Deciphering signatures of mutational processes operative in human cancer.
The genome of a cancer cell carries somatic mutations that are the cumulative consequences of the DNA damage and repair processes operative during the cellular lineage between the fertilized egg and the cancer cell. Remarkably, these mutational processes are poorly characterized. Global sequencing initiatives are yielding catalogs of somatic mutations from thousands of cancers, thus providing the unique opportunity to decipher the signatures of mutational processes operative in human cancer. However, until now there have been no theoretical models describing the signatures of mutational processes operative in cancer genomes and no systematic computational approaches are available to decipher these mutational signatures. Here, by modeling mutational processes as a blind source separation problem, we introduce a computational framework that effectively addresses these questions. Our approach provides a basis for characterizing mutational signatures from cancer-derived somatic mutational catalogs, paving the way to insights into the pathogenetic mechanism underlying all cancers.


Estimating influence of cofragmentation on peptide quantification and identification in iTRAQ experiments by simulating multiplexed spectra.
Isobaric tag-based quantification such as iTRAQ and TMT is a promising approach to mass spectrometry-based quantification in proteomics as it provides wide proteome coverage with greatly increased experimental throughput. However, it is known to suffer from inaccurate quantification and identification of a target peptide due to cofragmentation of multiple peptides, which likely leads to under-estimation of differentially expressed peptides (DEPs). A simple method of filtering out cofragmented spectra with less than 100% precursor isolation purity (PIP) would decrease the coverage of iTRAQ/TMT experiments. In order to estimate the impact of cofragmentation on quantification and identification of iTRAQ-labeled peptide samples, we generated multiplexed spectra with varying degrees of PIP by mixing the two MS/MS spectra of 100% PIP obtained in global proteome profiling experiments on gastric tumor-normal tissue pair proteomes labeled by 4-plex iTRAQ. Despite cofragmentation, the simulation experiments showed that more than 99% of multiplexed spectra with PIP greater than 80% were correctly identified by three different database search engines-MODa, MS-GF+, and Proteome Discoverer. Using the multiplexed spectra that have been correctly identified, we estimated the effect of cofragmentation on peptide quantification. In 74% of the multiplexed spectra, however, the cancer-to-normal expression ratio was compressed, and a fair number of spectra showed the "ratio inflation" phenomenon. On the basis of the estimated distribution of distortions on quantification, we were able to calculate cutoff values for DEP detection from cofragmented spectra, which were corrected according to a specific PIP and probability of type I (or type II) error. When we applied these corrected cutoff values to real cofragmented spectra with PIP larger than or equal to 70%, we were able to identify reliable DEPs by removing about 25% of DEPs, which are highly likely to be false positives. Our experimental results provide useful insight into the effect of cofragmentation on isobaric tag-based quantification methods. The simulation procedure as well as the corrected cutoff calculation method could be adopted for quantifying the effect of cofragmentation and reducing false positives (or false negatives) in the DEP identification with general quantification experiments based on isobaric labeling techniques.


[Choosing the correct benzodiazepine: mechanism of action and pharmacokinetics].
There is a discrepancy between the recommendation for caution and daily practice in the prescription of benzodiazepines. Although there is heterogeneity in the registered indications, all benzodiazepine agonists have almost the same mechanism of action. There are, however, substantial pharmacokinetic differences between individual benzodiazepine agonists. During short-term use of benzodiazepines, the elimination half-life is no measure of duration of action. Benzodiazepine lipophilicity determines the speed of action. If a rapid effect is desired, for instance in acute anxiety or agitation, then regarding oral medication the use of a lipophilic benzodiazepine such as diazepam is a rational choice. An accumulation factor can be used to estimate benzodiazepine accumulation during chronic use. In theory, accumulation does not occur with once-daily dosage of benzodiazepines that have an elimination half-life markedly shorter than 24 h, such as oxazepam, temazepam, and lorazepam.


Neonatal outcomes following planned preterm delivery in diabetic mothers.
Women with diabetes may need elective preterm delivery due to pregnancy or diabetes related complications. The aim of this study was to describe the neonatal outcomes arising from elective preterm delivery in diabetic women. Suitable patients were identified by the obstetric team at Hull Royal Infirmary Women and Children's Hospital and data was extracted from their case notes. 45 diabetic women with planned preterm delivery were identified within a set time frame, resulting in 48 babies. Of the 48 babies born, 47 survived. 36 out of 48 were delivered via caesarean section. Gestational ages ranged from 29+3 to 36+6 weeks, and 24 out of 48 (50%) had a birth weight greater than the 90th centile for gestational age.34 out of the 48 babies experienced some form of neonatal complication and were admitted to the neonatal unit. The median duration of stay in the neonatal unit was 7 days. 14 of the surviving neonates suffered from respiratory distress, although only 4 required surfactant therapy to regain respiratory function. However, the incidence of serious neonatal complications in those born after 34 weeks was shown to be low. Elective preterm delivery after 34 weeks had little effect on overall neonatal outcome. Therefore it could be proposed that elective preterm delivery after 34 weeks gestation may be an acceptable option in diabetic women if there are maternal or obstetric complications.


Statistical mechanics of helix bundles using a dynamic programming approach.
Despite much study, biomolecule folding cooperativity is not well understood. There are quantitative models for helix-coil transitions and for coil-to-globule transitions, but no accurate models yet treat both chain collapse and secondary structure formation together. We develop here a dynamic programming approach to statistical mechanical partition functions of foldamer chain molecules. We call it the ascending levels model. We apply it to helix-coil and helix-bundle folding and cooperativity. For 14- to 50-mer Baldwin peptides, the model gives good predictions for the heat capacity and helicity versus temperature and urea. The model also gives good fits for the denaturation of Oas's three-helix bundle B domain of protein A (F13W*) and synthetic protein alpha3C by temperature and guanidine. The model predicts the conformational distributions. It shows that these proteins fold with transitions that are two-state, although the transitions in the Baldwin helices are nearly higher order. The model shows that the recently developed three-helix bundle polypeptoids of Lee et al. fold anti-cooperatively, with a predicted value of DeltaHvH/DeltaHcal = 0.72. The model also predicts that two-helix bundles are unstable in proteins but stable in peptoids. Our dynamic programming approach provides a general way to explore cooperativity in complex foldable polymers.


Effects of histamine, H1- and H2-receptor antagonists on the activity of serotonergic neurons in the dorsal raphe nucleus.
We investigated the effects of histamine applied by microiontophoresis onto serotonin-containing (serotonergic) cells recorded extracellularly in the dorsal raphe nucleus of the rat. Application of histamine at low iontophoretic currents (1-5 nA) produced a rapid depression of the firing of all serotonergic neurons tested. The H1-receptor antagonists mepyramine and diphenhydramine were unable to attenuate the histamine-induced response. Antagonism of the effect of histamine by the iontophoretic application of the H2-receptor antagonists cimetidine and metiamide was not possible to evaluate since both were found to exert potent inhibitory effects by themselves. In contrast, the nonimidazole-derived H2-receptor antagonist ranitidine, which had no effect by itself, selectively antagonized the histamine-induced depression of neuronal activity. Histidine, 3-methylhistamine and a variety of histamine agonists selective for H1- or H2-receptors were unable to mimic the effect of histamine in dorsal raphe. Histamine's effects may, in part, be mediated at a gamma-aminobutyric acid receptor complex as the gamma-aminobutyric acid antagonists bicuculline and picrotoxin rapidly and reversibly antagonized both the histamine- and the cimetidine-induced depression of serotonin cell firing; the glycine antagonist strychnine selectively blocked the inhibitory effect of glycine without altering the histamine-induced response. These data show an inhibitory effect of histamine on serotonin-containing neurons in the dorsal raphe; this effect may be partially mediated at a subtype of H2-receptor. These data further indicate that the inhibitory effects of histamine and cimetidine observed in the dorsal raphe nucleus may result, in part, from an action directly or indirectly at a gamma-aminobutyric acid receptor complex.


Transcriptomic versus chromosomal prognostic markers and clinical outcome in uveal melanoma.
To compare a gene expression-based classifier versus the standard genetic prognostic marker, monosomy 3, for predicting metastasis in uveal melanoma. Gene expression profiling, fluorescence in situ hybridization (FISH), and array comparative genomic hybridization (aCGH) were done on 67 primary uveal melanomas. Clinical and pathologic prognostic factors were also assessed. Variables were analyzed by Cox proportional hazards, Kaplan-Meier analysis, sensitivity, specificity, positive and negative predictive value, and positive and negative likelihood ratios. The gene expression-based molecular classifier assigned 27 tumors to class 1 (low risk) and 25 tumors to class 2 (high risk). By Cox univariate proportional hazards, class 2 signature (P = 0.0001), advanced patient age (P = 0.01), and scleral invasion (P = 0.007) were the only variables significantly associated with metastasis. Only the class 2 signature was needed to optimize predictive accuracy in a Cox multivariate model. A less significant association with metastasis was observed for monosomy 3 detected by aCGH (P = 0.076) and FISH (P = 0.127). The sensitivity and specificity for the molecular classifier (84.6% and 92.9%, respectively) were superior to monosomy 3 detected by aCGH (58.3% and 85.7%, respectively) and FISH (50.0% and 72.7%, respectively). Positive and negative predictive values (91.7% and 86.7%, respectively) and positive and negative likelihood ratios (11.9 and 0.2, respectively) for the molecular classifier were also superior to those for monosomy 3. Molecular classification based on gene expression profiling of the primary tumor was superior to monosomy 3 and clinicopathologic prognostic factors for predicting metastasis in uveal melanoma.


Stereoscopic virtual realistic surgical simulation in intracranial aneurysms.
Three-dimensional (3D)-computed tomographic angiography (CTA) has been widely used for surgical simulation of intracranial aneurysms. Stereo imaging technology is progressing rapidly in recent years and stereo imaging may make more realistic surgical simulation possible. Therefore, we aimed at the establishment of a technique for stereoscopic viewing of minute volume rendering images while pursuing a low cost. Between January 2009 and June 2011, 54 patients with ruptured intracranial aneurysms were enrolled in this study. CTA data was transferred to the workstation equipped with image-processing software, and multilayer fusion images were processed by neurosurgeons. Image data for stereoscopic viewing of multilayer fusion image from arbitrary directions were collected form rotational trajectories around an aneurysm and were output to MPEG file. Stereoscopic viewing using MPEG data was achieved by the freeware named Stereo Movie Maker. Stereo viewing method using QuickTime VR format was also tried. Multilayer fusion image created from CTA data displayed clearly the anatomical information about not only the aneurysm but also the surrounding structures, such as parent artery, venous system, brain tissue, skull bone, and scalp. The quality of the resulting multilayer fusion image was suitable for surgical simulation with virtual reality. Virtual realistic surgical simulation became possible by the combination of minute multilayer fusion image and stereoscopic viewing by our method. Our method for stereo viewing of multilayer fusion images resulted in an improvement in the capability of diagnostic imaging and the image-guided support for neurosurgical procedures in intracranial aneurysm.


Structure and enzymatic accessibility of leaf and stem from wheat straw before and after hydrothermal pretreatment.
Biomass recalcitrance is affected by a number of chemical, physical and biological factors. In this study we looked into the differences in recalcitrance between two major anatomical fractions of wheat straw biomass, leaf and stem. A set of twenty-one wheat cultivars was fractionated and illustrated the substantial variation in leaf-to-stem ratio between cultivars. The two fractions were compared in terms of chemical composition, enzymatic convertibility, cellulose crystallinity and glucan accessibility. The use of water as a probe for assessing glucan accessibility was explored using low field nuclear magnetic resonance and infrared spectroscopy in combination with hydrogen-deuterium exchange. Leaves were clearly more degradable by lignocellulolytic enzymes than stems, and it was demonstrated that xylose removal was more linked to glucose yield for stems than for leaves. Comparing the locations of water in leaf and stem by low field NMR and FT-IR revealed that the glucan hydroxyl groups in leaves were more accessible to water than glucan hydroxyl groups in stems. No difference in crystallinity between leaf and stem was observed using wide angle x-ray diffraction. Hydrothermal pretreatment increased the accessibility towards water in stems but not in leaves. The results in this study indicate a correlation between the accessibility of glucan to water and to enzymes. Enzymatic degradability of wheat straw anatomical fractions can be indicated by the accessibility of the hydroxyl groups to water. This suggests that water may be used to assess glucan accessibility in biomass samples.


Application of COMPOCHIP microarray to investigate the bacterial communities of different composts.
A microarray spotted with 369 different 16S rRNA gene probes specific to microorganisms involved in the degradation process of organic waste during composting was developed. The microarray was tested with pure cultures, and of the 30,258 individual probe-target hybridization reactions performed, there were only 188 false positive (0.62%) and 22 false negative signals (0.07%). Labeled target DNA was prepared by polymerase chain reaction amplification of 16S rRNA genes using a Cy5-labeled universal bacterial forward primer and a universal reverse primer. The COMPOCHIP microarray was applied to three different compost types (green compost, manure mix compost, and anaerobic digestate compost) of different maturity (2, 8, and 16 weeks), and differences in the microorganisms in the three compost types and maturity stages were observed. Multivariate analysis showed that the bacterial composition of the three composts was different at the beginning of the composting process and became more similar upon maturation. Certain probes (targeting Sphingobacterium, Actinomyces, Xylella/Xanthomonas/Stenotrophomonas, Microbacterium, Verrucomicrobia, Planctomycetes, Low G + C and Alphaproteobacteria) were more influential in discriminating between different composts. Results from denaturing gradient gel electrophoresis supported those of microarray analysis. This study showed that the COMPOCHIP array is a suitable tool to study bacterial communities in composts.


Does selection against transcriptional interference shape retroelement-free regions in mammalian genomes?
Eukaryotic genomes are scattered with retroelements that proliferate through retrotransposition. Although retroelements make up around 40 percent of the human genome, large regions are found to be completely devoid of retroelements. This has been hypothesised to be a result of genomic regions being intolerant to insertions of retroelements. The inadvertent transcriptional activity of retroelements may affect neighbouring genes, which in turn could be detrimental to an organism. We speculate that such retroelement transcription, or transcriptional interference, is a contributing factor in generating and maintaining retroelement-free regions in the human genome. Based on the known transcriptional properties of retroelements, we expect long interspersed elements (LINEs) to be able to display a high degree of transcriptional interference. In contrast, we expect short interspersed elements (SINEs) to display very low levels of transcriptional interference. We find that genomic regions devoid of long interspersed elements (LINEs) are enriched for protein-coding genes, but that this is not the case for regions devoid of short interspersed elements (SINEs). This is expected if genes are subject to selection against transcriptional interference. We do not find microRNAs to be associated with genomic regions devoid of either SINEs or LINEs. We further observe an increased relative activity of genes overlapping LINE-free regions during early embryogenesis, where activity of LINEs has been identified previously. Our observations are consistent with the notion that selection against transcriptional interference has contributed to the maintenance and/or generation of retroelement-free regions in the human genome.


Minimally invasive method for the point-of-care quantification of lymphatic vessel function.
Current clinical methods for the evaluation of lymphatic vessel function, crucial for early diagnosis and evaluation of treatment response of several pathological conditions, in particular of postsurgical lymphedema, are based on complex and mainly qualitative imaging techniques. To address this unmet medical need, we established a simple strategy for the painless and quantitative assessment of cutaneous lymphatic function. We prepared a lymphatic-specific tracer formulation, consisting of the clinically approved near-infrared fluorescent dye, indocyanine green, and the solubilizing surfactant Kolliphor HS15. The tracer was noninvasively delivered to the dermal layer of the skin using MicronJet600 hollow microneedles, and the fluorescence signal decay at the injection site was measured over time using a custom-made, portable detection device. The decay rate of fluorescence signal in the skin was used as a direct measure of lymphatic vessel drainage function. With this method, we could quantify impaired lymphatic clearance in transgenic mice lacking dermal lymphatics and distinguish distinct lymphatic clearance patterns in pigs in different body locations and under manual stimulus. Overall, this method has the potential for becoming a noninvasive and quantitative clinical "office test" for lymphatic function assessment.


Genome-wide identification of neuropeptides and their receptor genes in Bemisia tabaci and their transcript accumulation change in response to temperature stresses.
Insect neuropeptides play an important role in regulating physiological functions such as growth, development, behavior and reproduction. We identified temperature-sensitive neuropeptides and receptor genes of the cotton whitefly, Bemisia tabaci. We identified 38 neuropeptide precursor genes and 35 neuropeptide receptors and constructed a phylogenetic tree using additional data from other insects. As temperature adaptability enables B. tabaci to colonize a diversity of habitats, we performed quantitative polymerase chain reaction with two temperature stresses (low = 4 °C and high = 40 °C) to screen for temperature-sensitive neuropeptides. We found many neuropeptides and receptors that may be involved in the temperature adaptability of B. tabaci. This study is the first to identify B. tabaci neuropeptides and their receptors, and it will help to reveal the roles of neuropeptides in temperature adaptation of B. tabaci.


A translation and preliminary validation of the Dutch Wound-QoL questionnaire.
Chronic wounds have a major impact on patients' health-related quality of life (HRQoL). Therefore, measuring HRQoL is an indispensable part of the treatment of patients with chronic wounds. The aim of this study was to translate and validate the Wound-QoL, a wound-specific HRQoL questionnaire, in a Dutch population. The Wound-QoL was translated into Dutch according to the international standards. Patients with chronic wounds were asked to complete questionnaires at baseline (T0) and after six weeks (T1), including Wound-QoL, EQ-5D-3L (a generic questionnaire to measure HRQoL) and a visual analogue scale (VAS) measuring wound pain. If patients were not able to complete the questionnaire by themselves, it was read out to them by a nurse. Further data were obtained from medical records. Of the 120 patients included, 64 (53.3%) completed the questionnaire by themselves. To 55 patients (45.8%), the questionnaire was read out. The internal consistency of the Wound-QoL global score was high at both time points (T0: Cronbach's α = 0.89, T1: Cronbach's α = 0.92). The item selectivity for global score ranged from r = 0.25 to r = 0.77 at T0 and from r = 0.40 to r = 0.79 at T1. Overall, the self-completion and read-out subgroups showed similar internal consistency and item selectivity scores. With regard to convergent validity, significant correlations were found between Wound-QoL and EQ-5D-3L (T0: r = - 0.45, p < 0.001, T1: r = - 0.50, p < 0.001) as well as between Wound-QoL and pain VAS (T0: r = 0.23, p = 0.012, T1: r = 0.37, p = 0.001) at both time points. Responsiveness analyses showed significant correlations between changes in Wound-QoL and changes in EQ-5D-3L (r = - 0.37, p < 0.001), pain VAS (r = 0.24, p = 0.044) and wound size (r = 0.24, p = 0.013). The self-completion and read-out subgroups showed differences in convergent validity and responsiveness. The results indicate that the Dutch version of the Wound-QoL has positive psychometric properties. However, more research is needed to further explore the differences between self-completed and read-out questionnaires.


Interleukin-21 contributes to germinal centre formation and immunoglobulin G4 production in IgG4-related dacryoadenitis and sialoadenitis, so-called Mikulicz's disease.
Interleukin (IL)-21 is mainly produced by CD4 T helper (Th) cells including Th2, Th17 and follicular helper T (Tfh) cells. Recent studies have reported that IL-21 is involved in the formation of germinal centres (GCs) and class switching of IgG4. It has been suggested that IgG4-related dacryoadenitis and sialoadenitis (IgG4-DS), so-called Mikulicz's disease (MD), is distinct from Sjögren's syndrome (SS) and shows a high frequency of GC formation in salivary glands. In this study the expression of IL-21 in IgG4-DS and SS patients was examined. Twelve patients with IgG4-DS, 15 with SS and 15 healthy subjects were screened for (1) ectopic GC formation in formalin-fixed labial salivary gland (LSG) biopsy samples; (2) expression of IL-21, Th2-, Th17- and Tfh-related molecules (cytokines, chemokine receptors and transcription factors) in LSGs; (3) relationship between IgG4/IgG ratio and mRNA expression of IL-21 in LSGs. mRNA expression of IL-21 and Bcl-6 in LSGs from patients with IgG4-DS was significantly higher than in patients with SS and controls. IL-21 and CXCR5 were detected by immunohistochemistry in or around GC in patients with SS and those with IgG4-DS. IL-21 was detected in infiltrating lymphocytes outside GC only in patients with IgG4-DS. Expression of IL-21 was consistent with that of Th2-related molecules while IL-17 was rarely seen in IgG4-DS. Furthermore, the expression of IL-21 in LSGs was correlated with the number of GC formations and the IgG4/IgG ratio in patients with IgG4-DS. These results suggest that overexpression of IL-21 by Th2 cells might play a key role in GC formation and IgG4 production in IgG4-DS.


Insulin-induced hexokinase II expression is reduced in obesity and NIDDM.
NIDDM and obesity are characterized by decreased insulin-stimulated glucose uptake in muscle. It has been suggested that impaired glucose phosphorylation to glucose-6-phosphate, catalyzed in muscle by hexokinase (HK)II, may contribute to this insulin resistance. Insulin is known to increase HKII mRNA, protein, and activity in lean nondiabetic individuals. The purpose of this study was to determine whether defects in insulin-stimulated HKII expression and activity could contribute to the insulin resistance of obesity and NIDDM. Fifteen lean nondiabetic control subjects, 17 obese nondiabetic subjects, and 14 obese NIDDM patients were studied. Percutaneous muscle biopsies of the vastus lateralis were performed in conjunction with leg balance and local indirect calorimetry measurements before and at the end of a 3-h euglycemic-hyperinsulinemic clamp (40 or 240 mU x min(-1) x m[-2]). Leg glucose uptake in response to the 40-mU insulin infusion was higher in the lean control subjects (2.53 +/- 0.35 micromol x min(-1) per x 100 ml leg vol) than in obese (1.46 +/- 0.50) or NIDDM (0.53 +/- 0.25, P < 0.05) patients. In response to 240 mU insulin, leg glucose uptake was similar in all of the groups. In response to 40 mU insulin, HKII mRNA in lean control subjects was increased 1.48 +/- 0.18-fold (P < 0.05) but failed to increase significantly in the obese (1.12 +/- 0.24) or NIDDM (1.14 +/- 0.18) groups. In response to 240 mU insulin, HKII mRNA was increased in all groups (control subjects 1.48 +/- 0.18, P < 0.05 vs. basal, obese 1.30 +/- 0.16, P < 0.05, and NIDDM 1.25 +/- 0.14, P < 0.05). Under basal conditions, HKI and HKII activities did not differ significantly between groups. Neither the 40 mU nor the 240 mU insulin infusion affected HK activity. Total HKII activity was reduced in the obese subjects (4.33 +/- 0.08 pmol x min(-1) x g(-1) muscle protein) relative to the lean control subjects (5.00 +/- 0.08, P < 0.05). There was a further reduction in the diabetic patients (3.10 +/- 0.10, P < 0.01 vs. the control subjects, P < 0.01 vs. the obese subjects). Resistance to insulin's metabolic effects extends to its ability to induce HKII expression in obesity and NIDDM.


Anaplastic, plasmablastic, and plasmacytic plasmacytomas of mice: relationships to human plasma cell neoplasms and late-stage differentiation of normal B cells.
We have compared histologic features and gene expression profiles of newly identified plasmacytomas from NFS.V(+) congenic mice with plasmacytomas of IL6 transgenic, Fasl mutant, and SJL-beta2M(-/-) mice. NFS.V(+) tumors comprised an overlapping morphologic spectrum of high-grade/anaplastic, intermediate-grade/plasmablastic, and low-grade/plasmacytic cases with similarities to subsets of human multiple myeloma and plasmacytoma. Microarray and immunohistochemical analyses of genes expressed by the most prevalent tumors, plasmablastic plasmacytomas, showed them to be most closely related to immunoblastic lymphomas, less so to plasmacytomas of Fasl mutant and SJL mice, and least to plasmacytic plasmacytomas of IL6 transgenic mice. Plasmablastic tumors seemed to develop in an inflammatory environment associated with gene signatures of T cells, natural killer cells, and macrophages not seen with plasmacytic plasmacytomas. Plasmablastic plasmacytomas from NFS.V(+) and SJL-beta2M(-/-) mice did not have structural alterations in Myc or T(12;15) translocations and did not express Myc at high levels, regular features of transgenic and pristane-induced plasmacytomas. These findings imply that, as for human multiple myeloma, Myc-independent routes of transformation contribute to the pathogenesis of these tumors. These findings suggest that plasma cell neoplasms of mice and humans exhibit similar degrees of complexity. Mouse plasmacytomas, previously considered to be homogeneous, may thus be as diverse as their human counterparts with respect to oncogenic mechanisms of plasma cell transformation. Selecting specific types of mouse plasmacytomas that relate most closely to subtypes of human multiple myeloma may provide new opportunities for preclinical testing of drugs for treatment of the human disease.


Genetic defects in pediatric-onset adrenal insufficiency in Japan.
Most patients with pediatric-onset primary adrenal insufficiency (PAI), such as 21-hydroxylase deficiency, can be diagnosed by measuring the urine or serum levels of steroid metabolites. However, the etiology is often difficult to determine in a subset of patients lacking characteristic biochemical findings. To assess the frequency of genetic defects in Japanese children with biochemically uncharacterized PAI and characterize the phenotypes of mutation-carrying patients. We enrolled 63 Japanese children (59 families) with biochemically uncharacterized PAI, and sequenced 12 PAI-associated genes. The pathogenicities of rare variants were assessed based on in silico analyses and structural modeling. We calculated the proportion of mutation-carrying patients according to demographic characteristics. We identified genetic defects in 50 (85%) families: STAR in 19, NR0B1 in 18, SAMD9 in seven, AAAS in two, NNT in two, MC2R in one and CDKN1C in one. NR0B1 defects were identified in 78% of the male patients that received both glucocorticoid and mineralocorticoid replacement therapy and had normal male external genitalia. STAR defects were identified in 67% of female and 9% of male patients. Seven of the 19 patients with STAR defects developed PAI at age two or older, out of whom, five did not have mineralocorticoid deficiency. Molecular testing elucidated the etiologies of most biochemically uncharacterized PAI patients. Genetic defects such as NR0B1 defects are presumed based on phenotypes, while others with broad phenotypic variability, such as STAR defects, are difficult to diagnose. Molecular testing is a rational approach to diagnosis in biochemically uncharacterized PAI patients.


Studies on the clinical significance of nonesterified and total cholesterol in urine.
Gas-liquid chromatographic determinations of nonesterified and total urinary cholesterol were performed in 137 normals, 264 patients with various internal diseases without evidence of neoplasias or diseases of the kidney or urinary tract, 497 patients with malignancies and 236 patients with diseases of the kidney, urinary tract infections or prostatic adenoma with residual urine. A normal range (mean +/- 2 SD) of 0.2-2.2 mg/24 hours nonesterified cholesterol (NEC) and of 0.3-3.0 mg/24 hours total cholesterol (TC) was calculated. Values of urinary cholesterol excretion were independent of age and sex and did not correlate with cholesterol levels in plasma. Patients with various internal diseases, without evidence of neoplasias nor diseases of the kidney or obstruction of the urinary tract, showed normal urinary cholesterol excretions, as did patients with infections of the urinary tract. However, elevated urinary cholesterol was found in patients with diseases of the kidney or urinary tract obstruction (prostatic adenoma with residual urine), malignant diseases of the urogenital tract and metastasizing carcinoma of the breast. In patients with other malignant diseases urinary cholesterol was usually normal. Lesions of the urothelial cell membranes are considered to be the most likely cause of urinary cholesterol hyperexcretion. The clinical value of urinary cholesterol determinations as a possible screening test for urogenital carcinomas in unselected populations is limited by lacking specificity, expensive methodology and low prevalence of the mentioned carcinomas, although elevated urinary cholesterol excretions have been observed in early clinical stages of urogenital cancers.


Target loop adaptive optics: thermal blooming correction.
To evaluate adaptive optics (AO) concepts for the correction of nonlinear atmospheric phenomena occurring in high energy laser beam propagation and to perform AO system trade offs, a simple, but reasonably accurate, model of the entire AO loop is required. Heretofore, no such model existed which takes into account the temporal response of the atmosphere and yet is simple enough to permit extensive system tradeoffs. We have developed such a model. Optimum thermal blooming correction dictates that the atmospheric response time must be accounted for properly, especially for strong blooming strength. Experimental verification of the above model was carried out using an argon-ion laser with an iodine blooming cell and a closed AO loop to provide corrections for tilt, focus, and astigmatism. Reasonable agreement between theory and experimental data was achieved.


Predictive Modeling of Anatomy with Genetic and Clinical Data.
We present a semi-parametric generative model for predicting anatomy of a patient in subsequent scans following a single baseline image. Such predictive modeling promises to facilitate novel analyses in both voxel-level studies and longitudinal biomarker evaluation. We capture anatomical change through a combination of population-wide regression and a non-parametric model of the subject's health based on individual genetic and clinical indicators. In contrast to classical correlation and longitudinal analysis, we focus on predicting new observations from a single subject observation. We demonstrate prediction of follow-up anatomical scans in the ADNI cohort, and illustrate a novel analysis approach that compares a patient's scans to the predicted subject-specific healthy anatomical trajectory.


Efficacy of a trivalent influenza vaccine against seasonal strains and against 2009 pandemic H1N1: A randomized, placebo-controlled trial.
Before pandemic H1N1 vaccines were available, the potential benefit of existing seasonal trivalent inactivated influenza vaccines (IIV3s) against influenza due to the 2009 pandemic H1N1 influenza strain was investigated, with conflicting results. This study assessed the efficacy of seasonal IIV3s against influenza due to 2008 and 2009 seasonal influenza strains and against the 2009 pandemic H1N1 strain. This observer-blind, randomized, placebo-controlled study enrolled adults aged 18-64years during 2008 and 2009 in Australia and New Zealand. Participants were randomized 2:1 to receive IIV3 or placebo. The primary objective was to demonstrate the efficacy of IIV3 against laboratory-confirmed influenza. Participants reporting an influenza-like illness during the period from 14days after vaccination until 30 November of each study year were tested for influenza by real-time reverse transcription polymerase chain reaction. Over a study period of 2years, 15,044 participants were enrolled (mean age±standard deviation: 35.5±14.7years; 54.4% female). Vaccine efficacy of the 2008 and 2009 IIV3s against influenza due to any strain was 42% (95% confidence interval [CI]: 30%, 52%), whereas vaccine efficacy against influenza due to the vaccine-matched strains was 60% (95% CI: 44%, 72%). Vaccine efficacy of the 2009 IIV3 against influenza due to the 2009 pandemic H1N1 strain was 38% (95% CI: 19%, 53%). No vaccine-related deaths or serious adverse events were reported. Solicited local and systemic adverse events were more frequent in IIV3 recipients than placebo recipients (local: IIV3 74.6% vs placebo 20.4%, p<0.001; systemic: IIV3 46.6% vs placebo 39.1%, p<0.001). The 2008 and 2009 IIV3s were efficacious against influenza due to seasonal influenza strains and the 2009 IIV3 demonstrated moderate efficacy against influenza due to the 2009 pandemic H1N1 strain. Funded by CSL Limited, ClinicalTrials.gov identifier NCT00562484.


Ion-regulated allosteric binding of fullerenes (C60 and C70) by tetrathiafulvalene-calix[4]pyrroles.
The effect of ionic species on the binding of fullerenes (C60 and C70) by tetrathiafulvalene-calix[4]pyrrole (TTF-C4P) receptors and the nature of the resulting supramolecular complexes (TTF-C4P + fullerene + halide anion + tetraalkylammonium cation) was studied in the solid state through single crystal X-ray diffraction methods and in dichloromethane solution by means of continuous variation plots and UV-vis spectroscopic titrations. These analyses revealed a 1:1 stoichiometry between the anion-bound TTF-C4Ps and the complexed fullerenes. The latter guests are bound within the bowl-like cup of the C4P in a ball-and-socket binding mode. The interactions between the TTF-C4P receptors and the fullerene guests are highly influenced by both the nature of halide anions and their counter tetraalkylammonium cations. Three halides (F(-), Cl(-), and Br(-)) were studied. All three potentiate the binding of the two test fullerenes by inducing a conformational change from the 1,3-alternate to the cone conformer of the TTF-C4Ps, thus acting as positive heterotropic allosteric effectors. For a particular halide anion, the choice of tetraalkylammonium salts serves to modulate the strength of the TTF-C4P-fullerene host-guest binding interactions and, in conjunction with variations in the halide anion, can be exploited to alter the inherent selectivity of the host for a given fullerene. Differences in binding are reflected in the excited state optical properties. Overall, the present four-component system provides an illustration of how host-guest binding events involving appropriately designed artificial receptors can be fine-tuned via the addition of simple ionic species as allosteric modulators.


Effectiveness of medical and surgical therapies for lower urinary tract symptoms in the community setting.
Study Type - Therapy (outcomes research) Level of Evidence 2c. What's known on the subject? and What does the study add? It is known that benign prostatic hyperplasia is a common condition affecting most men by the age of 80 years. There are multiple treatment options available, including both medical and surgical interventions. However, what is not known is how affective the different types of interventions are in the general population. Previous studies have focused on centre-specific data. What is unique about our study is that it is a prospective cross-section analysis of a community cohort of men. Through this study we were able to assess the outcomes in the general population as opposed to in a high-volume surgical centre. Our findings show that in this community medical management was poor at symptomatic improvement, whereas surgical intervention produced the best improvement. • To describe the use and symptomatic outcomes of different therapies for lower urinary tract symptoms (LUTS) in a community-based population of men followed for 17 years. • Data from a randomly selected cohort of 2184 men, aged 40-79 years in 1990, from Olmsted County, Minnesota, USA were included in the study. Participants completed a questionnaire similar to the American Urological Association Symptom Index (AUASI) and reported on incontinence. • Men were followed biennially through 2007 (median follow-up: 13.7 years; Q1, Q3: 8.8, 15.7). Medical and surgical treatments for LUTS were reported on biennial questionnaires and abstracted from community medical records. • Overall, 610 (28%) men received medical or surgical therapy for treatment of LUTS. Patients undergoing vaporization and transurethral resection of the prostate (TURP) had the highest pre-intervention AUASI scores (P < 0.001) and the most rapid increase in scores over time (P= 0.002) compared with those treated with medications or no therapy. After intervention, symptom progression slowed in all treatment groups. • However, the greatest improvement in AUASI score (median % change) was observed in the TURP group: -27.45%. The TURP group also reported a significant decrease in incontinence after surgery (% change): TURP: -22.58%. • All therapies were effective at slowing the progression of LUTS, but only TURP patients reported a significant decrease in both LUTS and incontinence after therapy.


Generation and characterization of an antagonistic monoclonal antibody against an extracellular domain of mouse DP2 (CRTH2/GPR44) receptors for prostaglandin D2.
Prostaglandin D2 (PGD2) is a lipid mediator involved in sleep regulation and inflammation. PGD2 interacts with 2 types of G protein-coupled receptors, DP1 and DP2/CRTH2 (chemoattractant receptor homologous molecule expressed on T helper type 2 cells)/GPR44 to show a variety of biological effects. DP1 activation leads to Gs-mediated elevation of the intracellular cAMP level, whereas activation of DP2 decreases this level via the Gi pathway; and it also induces G protein-independent, arrestin-mediated cellular responses. Activation of DP2 by PGD2 causes the progression of inflammation via the recruitment of lymphocytes by enhancing the production of Th2-cytokines. Here we developed monoclonal antibodies (MAbs) against the extracellular domain of mouse DP2 by immunization of DP2-null mutant mice with DP2-overexpressing BAF3, murine interleukin-3 dependent pro-B cells, to reduce the generation of antibodies against the host cells by immunization of mice. Moreover, we immunized DP2-KO mice to prevent immunological tolerance to mDP2 protein. After cell ELISA, immunocytochemical, and Western blot analyses, we successfully obtained a novel monoclonal antibody, MAb-1D8, that specifically recognized native mouse DP2, but neither human DP2 nor denatured mouse DP2, by binding to a particular 3D receptor conformation formed by the N-terminus and extracellular loop 1, 2, and 3 of DP2. This antibody inhibited the binding of 0.5 nM [3H]PGD2 to mouse DP2 (IC50 = 46.3 ± 18.6 nM), showed antagonistic activity toward 15(R)-15-methyl PGD2-induced inhibition of 300 nM forskolin-activated cAMP production (IC50 = 16.9 ± 2.6 nM), and gave positive results for immunohistochemical staining of DP2-expressing CD4+ Th2 lymphocytes that had accumulated in the kidney of unilateral ureteral obstruction model mice. This monoclonal antibody will be very useful for in vitro and in vivo studies on DP2-mediated diseases.


Beyond aflatoxin: four distinct expression patterns and functional roles associated with Aspergillus flavus secondary metabolism gene clusters.
Species of Aspergillus produce a diverse array of secondary metabolites, and recent genomic analysis has predicted that these species have the capacity to synthesize many more compounds. It has been possible to infer the presence of 55 gene clusters associated with secondary metabolism in Aspergillus flavus; however, only three metabolic pathways-aflatoxin, cyclopiazonic acid (CPA) and aflatrem-have been assigned to these clusters. To gain an insight into the regulation of and to infer the ecological significance of the 55 secondary metabolite gene clusters predicted in A. flavus, we examined their expression over 28 diverse conditions. Variables included culture medium and temperature, fungal development, colonization of developing maize seeds and misexpression of laeA, a global regulator of secondary metabolism. Hierarchical clustering analysis of expression profiles allowed us to categorize the gene clusters into four distinct clades. Gene clusters for the production of aflatoxins, CPA and seven other unknown compound(s) were identified as belonging to one clade. To further explore the relationships found by gene expression analysis, aflatoxin and CPA production were quantified under five different cell culture environments known to be conducive or nonconducive for aflatoxin biosynthesis and during the colonization of developing maize seeds. Results from these studies showed that secondary metabolism gene clusters have distinctive gene expression profiles. Aflatoxin and CPA were found to have unique regulation, but are sufficiently similar that they would be expected to co-occur in substrates colonized with A. flavus.


Environmental risks and children's health: what can PRAMS tell us?
Environmental exposures during pregnancy have a lasting impact on children's health. We combined environmental and maternal risk factor survey data to inform efforts to protect children's health. We made recommendations for future use of such data. A modified version of the Pregnancy Risk Assessment Monitoring System (PRAMS) mail survey was conducted based on weighted sampling design with low-income and non-low income women in Monroe County, NY (1,022 respondents). A series of environmental questions were included in the questionnaire. Data were analyzed using Chi square tests and Poisson loglinear regression model to identify patterns in environmental health risk and sociodemographic characteristics. We identified women who rented their homes, had lower incomes, and lived in inner city zip codes as "high environmental health risk" (HEHR). HEHR respondents were more likely to report that a health care provider talked with them about lead and on average reported more behaviors to protect their children from lead poisoning. Combining environmental and perinatal risk factor data could yield important recommendations for medical practice, health education, and policy development. However, at present PRAMS gathers only limited and inconsistent environmental data. We found that existing PRAMS environmental questions are insufficient. Further work is needed to develop updated and more comprehensive environmental health survey questions and implement them consistently across the country.


Heteroatom-doped carbon dots: synthesis, characterization, properties, photoluminescence mechanism and biological applications.
Heteroatom-doped carbon dots (CDs), due to their excellent photoluminescence (PL) properties, attracted widespread attention recently and demonstrated immense promise for diverse applications, particularly for biological applications. The objective of this feature article is to provide a comprehensive overview of the recent progress in the research and development of heteroatom-doped CDs and a detailed description of the influence of single or co-doping heteroatoms on their PL behavior. The most recent understanding and critical insights into the PL mechanism of heteroatom-doped CDs are also highlighted. Moreover, potential bio-related applications of heteroatom-doped CDs in biosensing, bioimaging, and theranostics are also reviewed. This state-of-the-art review will provide a platform for understanding the intricate details of heteroatom-doped CDs, a summary of the latest progress in the field, and related applications in biology and is expected to inspire further developments in this exciting class of materials.


Heparin-induced tau filaments are structurally heterogeneous and differ from Alzheimer's disease filaments.
Alzheimer's disease (AD) is characterized by the presence of tau filaments in the brain whose structure was recently solved. The formation of AD filaments is routinely modeled in vitro by mixing tau with heparin. This study shows that heparin-induced tau filaments are markedly different from the AD filaments and are highly heterogeneous.


Discovery and Structure-Activity Relationship Study of (Z)-5-Methylenethiazolidin-4-one Derivatives as Potent and Selective Pan-phosphatidylinositol 5-Phosphate 4-Kinase Inhibitors.
Due to their role in many important signaling pathways, phosphatidylinositol 5-phosphate 4-kinases (PI5P4Ks) are attractive targets for the development of experimental therapeutics for cancer, metabolic, and immunological disorders. Recent efforts to develop small molecule inhibitors for these lipid kinases resulted in compounds with low- to sub-micromolar potencies. Here, we report the identification of CVM-05-002 using a high-throughput screen of PI5P4Kα against our in-house kinase inhibitor library. CVM-05-002 is a potent and selective inhibitor of PI5P4Ks, and a 1.7 Å X-ray structure reveals its binding interactions in the ATP-binding pocket. Further investigation of the structure-activity relationship led to the development of compound 13, replacing the rhodanine-like moiety present in CVM-05-002 with an indole, a potent pan-PI5P4K inhibitor with excellent kinome-wide selectivity. Finally, we employed isothermal cellular thermal shift assays (CETSAs) to demonstrate the effective cellular target engagement of PI5P4Kα and -β by the inhibitors in HEK 293T cells.


Speed of language comprehension is impaired in ADHD.
Children with ADHD have an increased risk of poor academic performance. It is important to identify cognitive processes that may be related to this academic failure. In Western schooling systems, especially language processing skills may be of relevance. The present study, therefore, compares the ability to comprehend complex sentences of individuals with and without ADHD. Fifteen children (aged 8-11) and 15 adolescents (aged 12-16) with ADHD combined subtype are matched for age, gender, and parental level of education to 30 control subjects. Language comprehension is measured using the neuropsychological procedure proposed by Luria and an adapted version of the Token Test. Compared with the control group, children and adolescents with ADHD perform significantly slower on language comprehension tasks. Differences in accuracy are limited. No interaction between age and ADHD is found. Children and adolescents with ADHD are slower and less efficient than matched control subjects with regard to complex sentence comprehension.


Esophageal early basaloid squamous carcinoma with unusual narrowband imaging magnified endoscopy findings.
Basaloid squamous carcinoma (BSC) is a rare variant of esophageal cancer. There are very few reports of "early" BSC. Here we report a case of early BSC with unusual findings by narrowband imaging magnified endoscopy (NBI-ME). A 70-year-old man with a middle thoracic esophageal tumor was referred to our hospital. White-light endoscopy revealed a reddish depressed lesion 5 mm in diameter having a subepithelial tumor-like prominence with a gentle rising slope. NBI-ME revealed irregular loop-shaped microvessels coexistent with thick irregularly branched non-looped vessels. Iodine staining revealed a pale brown lesion. We performed endoscopic submucosal dissection for diagnostic treatment. Histologic examination showed the proliferation of basal cell-like hyperchromatic tumor cells in the lamina propria and with slight invasion into the submucosa at a depth of 320 μm. The tumor cells formed solid nests and microcystic structures, containing an Alcian blue-positive mucoid matrix. The surface was covered with squamous epithelium without cellular atypia. Thin vessels were observed in the intra-epithelial papilla and thick vessels were observed around the solid nests beneath the epithelium. Based on these findings together, we diagnosed the lesion as BSC. In this case, the NBI-ME findings differed from those of typical squamous cell carcinoma in that both non-invasive cancer-like irregular loop-shaped microvessels coexisted with massively invasive cancer-like thick non-looped vessels. We speculate that the looped and non-looped vessels observed by NBI-ME histologically corresponded to thin vessels in the intra-epithelial papilla and thick vessels around the tumor nests, respectively. These NBI-ME findings might be a feature of early esophageal BSC.


Birdsong "transcriptomics": neurochemical specializations of the oscine song system.
Vocal learning is a rare and complex behavioral trait that serves as a basis for the acquisition of human spoken language. In songbirds, vocal learning and production depend on a set of specialized brain nuclei known as the song system. Using high-throughput functional genomics we have identified approximately 200 novel molecular markers of adult zebra finch HVC, a key node of the song system. These markers clearly differentiate HVC from the general pallial region to which HVC belongs, and thus represent molecular specializations of this song nucleus. Bioinformatics analysis reveals that several major neuronal cell functions and specific biochemical pathways are the targets of transcriptional regulation in HVC, including: 1) cell-cell and cell-substrate interactions (e.g., cadherin/catenin-mediated adherens junctions, collagen-mediated focal adhesions, and semaphorin-neuropilin/plexin axon guidance pathways); 2) cell excitability (e.g., potassium channel subfamilies, cholinergic and serotonergic receptors, neuropeptides and neuropeptide receptors); 3) signal transduction (e.g., calcium regulatory proteins, regulators of G-protein-related signaling); 4) cell proliferation/death, migration and differentiation (e.g., TGF-beta/BMP and p53 pathways); and 5) regulation of gene expression (candidate retinoid and steroid targets, modulators of chromatin/nucleolar organization). The overall direction of regulation suggest that processes related to cell stability are enhanced, whereas proliferation, growth and plasticity are largely suppressed in adult HVC, consistent with the observation that song in this songbird species is mostly stable in adulthood. Our study represents one of the most comprehensive molecular genetic characterizations of a brain nucleus involved in a complex learned behavior in a vertebrate. The data indicate numerous targets for pharmacological and genetic manipulations of the song system, and provide novel insights into mechanisms that might play a role in the regulation of song behavior and/or vocal learning.


A human antibody against Zika virus crosslinks the E protein to prevent infection.
The recent Zika virus (ZIKV) epidemic has been linked to unusual and severe clinical manifestations including microcephaly in fetuses of infected pregnant women and Guillian-Barré syndrome in adults. Neutralizing antibodies present a possible therapeutic approach to prevent and control ZIKV infection. Here we present a 6.2 Å resolution three-dimensional cryo-electron microscopy (cryoEM) structure of an infectious ZIKV (strain H/PF/2013, French Polynesia) in complex with the Fab fragment of a highly therapeutic and neutralizing human monoclonal antibody, ZIKV-117. The antibody had been shown to prevent fetal infection and demise in mice. The structure shows that ZIKV-117 Fabs cross-link the monomers within the surface E glycoprotein dimers as well as between neighbouring dimers, thus preventing the reorganization of E protein monomers into fusogenic trimers in the acidic environment of endosomes.