Optimization of a genotyping scheme for Treponema pallidum subsp. pallidum including markers for macrolide resistance

Syfilis er en seksuelt overførbar infeksjon, og er et viktig folkehelseproblem både i Norge og i verden. Forekomst av syfilis har økt i de senere årene, spesielt blant menn som har sex med menn (MSM). Bakterien Treponema pallidum subspecies pallidum (TPA), en spiroket som ikke lar seg dyrke, er årsaken til syfilis. Diagnostisering av syfilis gjøres ved hjelp av antistofftest mot TPA antigen, eller ved direkte identifisering av TPA DNA med polymerasekjedereaksjon (PCR). Ubehandlet syfilisinfeksjon, kan deles inn i ulike stadier som gir forskjellige symptomer: primær, sekundær, latent og tertiær. Det er ikke utviklet noen vaksine mot TPA. Penicillin er førstevalg ved behandling av syfilis. Azitromycin, et makrolidantibiotikum, har også vært anvendt i behandling av syfilis og brukes også i behandling av andre kjønnssykdommer. Resistens mot makrolidantibiotika i TPA har imidlertid blitt detektert globalt. TPA kan deles inn i to hovedgrupper utfra molekylær subtyping: Nichols- og SS14- stammer. Kombinasjonen av molekylær genotyping av TPA og resistens mot makrolidantibiotika er viktig for å forstå epidemiologien ved syfilis i Norge, og overvåkning av antibiotikaresistens. Genotypiskbestemmelse av TPA, inkludert markører for makrolidresistens, må først optimaliseres og etableres. Molekylær subtyping av TPA utføres ved multilokus sekvens typing (MLST) av tre loci: TP0136, TP0548 og TP0705. MLST består av en nested PCR etterfulgt av Sanger sekvensering for hvert locus. Allele profiler og sekvenstyper blir bestemt av PubMLST. For deteksjon av makrolidresistente TPA, som skyldes punktmutasjonene A2058G eller A2059G i ett eller begge kopier av 23S rRNA-genet, ble det utviklet en SimpleProbe real-time PCR analyse etterfulgt av en smeltepunktsanalyse. Makrolidresistente TPA har en lavere smeltepunkts-temperatur enn villtype TPA. Det ble samlet inn 81 pasientprøver fra oktober 2020 til september 2022. Ved molekylær subtyping ble det avdekket ni forskjellige sekvenstyper, deriblant tre nye sekvenstyper. Makrolidresistens kan også bekreftes blant disse prøvene.Syphilis, a sexually transmitted disease, is an important public health problem, both in Norway and worldwide. The incidence of syphilis has increased in the recent years, particularly among men who have sex with men (MSM). Infection with the uncultivated spirochaete bacteria Treponema pallidum subsp. pallidum (TPA), is the cause of syphilis. Syphilis can be diagnosed with antibody test against TPA antigen or by directly identification of TPA DNA with polymerase chain reaction (PCR). Untreated syphilis infection can be divided into different stages that give different symptoms: primary, secondary, latent, and tertiary. No vaccine against TPA has been developed yet. Penicillin is the first choice of antibiotic treatment for syphilis. Azithromycin, a macrolide antibiotic, has shown to be an effective treatment of early syphilis in many cases as well as treating infections with other venereal diseases. However, macrolide antibiotic resistance in TPA has been detected globally. By molecular characterization, TPA can be divided into two main lineages: Nichols- and SS14-like strains. The combination of molecular genotyping of TPA and macrolide antibiotic resistance is important for understanding the epidemiology of syphilis in Norway, and the surveillance of antibiotic resistance. Thus, a genotyping scheme for TPA including markers for macrolide resistance, need to be established and optimized. Molecular characterization of TPA was performed by multi-locus sequence typing (MLST) of three loci: TP0136, TP0548, and TP0705. MLST consists of a nested PCR followed by Sanger sequencing of each of the three loci. The allelic profiles and sequence type can be determined by PubMLST. For detection of macrolide resistance in TPA, due to point mutations in position A2058G or A2059G in one or both copies of the 23S rRNA gene, a SimpleProbe real-time PCR assay was developed followed by a melting point analysis. Macrolide resistance in TPA is detected by having a lower melting temperature than the wild-type. Eighty-one clinical samples were collected from October 2020 to September 2022. Molecular characterization of TPA revealed nine different sequence types, including three novel sequence types. Macrolide resistance was also confirmed among the collected samples

Enhancing Leisure Participation for Women who are Experiencing Homelessness

Opportunities to engage in leisure activities are limited for women experiencing homelessness (Klitzing, 2004). The purpose of this capstone was to develop a program for a residence serving women who are at risk of homelessness to improve their leisure participation and exploration. The needs assessment involved on-site observations at the Community Mission of Hope and Empowerment Village, literature review, client interviews, and an online questionnaire for volunteers and staff members. Although the program was not implemented due to time constraints and current programming already occurring, future sessions were outlined, and supplies were ordered through a grant to prepare for implementation. The 9-session program was based on the interests of the women at the Empowerment Village, incorporates aspects of wellness, and was proposed to promote more leisure opportunities for the women at the residence.https://soar.usa.edu/otdcapstonesfall2022/1001/thumbnail.jp

An exploratory trial implementing a community-based child oral health promotion intervention for Australian families from refugee and migrant backgrounds: a protocol paper for Teeth Tales

Introduction: Inequalities are evident in early childhood caries rates with the socially disadvantaged experiencing greater burden of disease. This study builds on formative qualitative research, conducted in the Moreland/Hume local government areas of Melbourne, Victoria 2006–2009, in response to community concerns for oral health of children from refugee and migrant backgrounds. Development of the community-based intervention described here extends the partnership approach to cogeneration of contemporary evidence with continued and meaningful involvement of investigators, community, cultural and government partners. This trial aims to establish a model for child oral health promotion for culturally diverse communities in Australia.<p></p> Methods and analysis: This is an exploratory trial implementing a community-based child oral health promotion intervention for Australian families from refugee and migrant backgrounds. Families from an Iraqi, Lebanese or Pakistani background with children aged 1–4 years, residing in metropolitan Melbourne, were invited to participate in the trial by peer educators from their respective communities using snowball and purposive sampling techniques. Target sample size was 600. Moreland, a culturally diverse, inner-urban metropolitan area of Melbourne, was chosen as the intervention site. The intervention comprised peer educator led community oral health education sessions and reorienting of dental health and family services through cultural Competency Organisational Review (CORe).<p></p> Ethics and dissemination: Ethics approval for this trial was granted by the University of Melbourne Human Research Ethics Committee and the Department of Education and Early Childhood Development Research Committee. Study progress and output will be disseminated via periodic newsletters, peer-reviewed research papers, reports, community seminars and at National and International conferences.<p></p&gt

Interictal Neurocognitive Processing of Visual Stimuli in Migraine: Evidence from Event-Related Potentials

Research has established decreased sensory habituation as a defining feature in migraine, while decreased cognitive habituation has only been found with regard to cognitive assessment of the relative probability of the occurrence of a stimulus event. Our study extended the investigation of interictal habituation in migraine to include cognitive processing when viewing of a series of visually-complex images, similar to those we encounter on the internet everyday. We examined interictal neurocognitive function in migraine from a habituation perspective, using a novel paradigm designed to assess how the response to a series of images changes over time. Two groups of participants--migraineurs (N = 25) and non-migraine controls (N = 25)--were asked to view a set of 232 unfamiliar logos in the context of a target identification task as their brain electrical responses were recorded via event-related potentials (ERPs). The set of logos was viewed serially in each of 10 separate trial blocks, with data analysis focusing on how the ERP responses to the logos in frontal electrodes from 200-600 ms changed across time within each group. For the controls, we found that the amplitude of the late positive potential (LPP) ERP component elicited by the logos had no significant change across trial blocks. In contrast, in migraineurs we found that the LPP significantly increased in amplitude across trial blocks, an effect consistent with a lack of habituation to visual stimuli seen in previous research. Our findings provide empirical support abnormal cognitive processing of complex visual images across time in migraineurs that goes beyond the sensory-level habituation found in previous research

A systematic review of measures of HIV/AIDS stigma in paediatric HIV-infected and HIV-affected populations

Introduction: HIV-related stigma impacts the quality of life and care management of HIV-infected and HIV-affected individuals, but how we measure stigma and its impact on children and adolescents has less often been described. Methods: We conducted a systematic review of studies that measured HIV-related stigma with a quantitative tool in paediatric HIV-infected and HIV-affected populations. Results and discussion: Varying measures have been used to assess stigma in paediatric populations, with most studies utilizing the full or variant form of the HIV Stigma Scale that has been validated in adult populations and utilized with paediatric populations in Africa, Asia and the United States. Other common measures included the Perceived Public Stigma Against Children Affected by HIV, primarily utilized and validated in China. Few studies implored item validation techniques with the population of interest, although scales were used in a different cultural context from the origin of the scale. Conclusions: Many stigma measures have been used to assess HIV stigma in paediatric populations, globally, but few have implored methods for cultural adaptation and content validity

The E3 Ubiquitin Ligase SCF(Cyclin F) Transmits AKT Signaling to the Cell-Cycle Machinery

The oncogenic AKT kinase is a key regulator of apoptosis, cell growth, and cell-cycle progression. Despite its important role in proliferation, it remains largely unknown how AKT is mechanistically linked to the cell cycle. We show here that cyclin F, a substrate receptor F-box protein for the SCF (Skp1/Cul1/F-box) family of E3 ubiquitin ligases, is a bona fide AKT substrate. Cyclin F expression oscillates throughout the cell cycle, a rare feature among the 69 human F-box proteins, and all of its known substrates are involved in proliferation. AKT phosphorylation of cyclin F enhances its stability and promotes assembly into productive E3 ligase complexes. Importantly, expression of mutant versions of cyclin F that cannot be phosphorylated by AKT impair cell-cycle entry. Our data suggest that cyclin F transmits mitogen signaling through AKT to the core cell-cycle machinery. This discovery has potential implications for proliferative control in malignancies where AKT is activated

Transcriptome analysis of Drosophila suzukii reveals molecular mechanisms conferring pyrethroid and spinosad resistance.

Drosophila suzukii lay eggs in soft-skinned, ripening fruits, making this insect a serious threat to berry production. Since its 2008 introduction into North America, growers have used insecticides, such as pyrethroids and spinosads, as the primary approach for D. suzukii management, resulting in development of insecticide resistance in this pest. This study sought to identify the molecular mechanisms conferring insecticide resistance in these populations. We sequenced the transcriptomes of two pyrethroid- and two spinosad-resistant isofemale lines. In both pyrethroid-resistant lines and one spinosad-resistant line, we identified overexpression of metabolic genes that are implicated in resistance in other insect pests. In the other spinosad-resistant line, we observed an overexpression of cuticular genes that have been linked to resistance. Our findings enabled the development of molecular diagnostics that we used to confirm persistence of insecticide resistance in California, U.S.A. To validate these findings, we leveraged D. melanogaster mutants with reduced expression of metabolic or cuticular genes that were found to be upregulated in resistant D. suzukii to demonstrate that these genes are involved in promoting resistance. This study is the first to characterize the molecular mechanisms of insecticide resistance in D. suzukii and provides insights into how current management practices can be optimized

The interactions of exogenous phytase with whole grain feeding and effects of barley as the whole grain component in broiler diets based on wheat, sorghum and wheat-sorghum blends

The objectives of this experiment were two-fold; the first was to evaluate exogenous phytase in either conventional or whole grain diets as a 2 × 2 factorial treatment array. Wheat-sorghum blended rations containing 12.5% ground or whole barley were offered without and with 1000 FTU/kg exogenous phytase. The second objective was to evaluate barley as the whole grain component in diets based on wheat, sorghum and equal wheat-sorghum blends as a 3 × 2 factorial treatment array. Rations based on wheat, sorghum and wheat-sorghum blends were offered as an intact pellet containing 12.5% ground barley or offered as a mix of 12.5% whole barley and a pelleted concentrate. Each of the dietary treatments was offered to 7 replicates (6 birds per cage) of male Ross 308 chicks from 7 to 28 days post-hatch. Treatment effects on growth performance, gizzard and pancreas weights, gizzard pH, bone mineralisation, nutrient utilisation, digestibility coefficients of starch and protein (N) and starch:protein disappearance rate ratios in four small intestinal segments (proximal and distal jejunum, proximal and distal ileum), excreta dry matter and incidence of dilated proventriculi were determined. In the 2 × 2 analysis there was a significant (

Long-Term Signs of T Cell and Myeloid Cell Activation After Intestinal Transplantation With Cellular Rejections Contributing to Further Increase of CD16+ Cell Subsets

The intestine mediates a delicate balance between tolerogenic and inflammatory immune responses. The continuous pathogen encounter might also augment immune cell responses contributing to complications observed upon intestinal transplantation (ITx). We thus hypothesized that ITx patients show persistent signs of immune cell activation affecting both the adaptive and innate immune cell compartment. Information on the impact of intestinal grafts on immune cell composition, however, especially in the long-term is sparse. We here assessed activated and differentiated adaptive and innate immune subsets according to time, previous experience of cellular or antibody-mediated rejections or type of transplant after ITx applying multi-parametric flow cytometry, gene expression, serum cytokine and chemokine profiling. ITx patients showed an increase in CD16 expressing monocytes and myeloid dendritic cells (DCs) compared to healthy controls. This was even detectable in patients who were transplanted more than 10 years ago. Also, conventional CD4+ and CD8+ T cells showed persistent signs of activation counterbalanced by increased activated CCR4+ regulatory T cells. Patients with previous cellular rejections had even higher proportions of CD16+ monocytes and DCs, whereas transplanting higher donor mass with multi-visceral grafts was associated with increased T cell activation. The persistent inflammation and innate immune cell activation might contribute to unsatisfactory results after ITx

Hierarchical Event Descriptor library schema for EEG data annotation

Standardizing terminology to describe electrophysiological events can improve both clinical care and computational research. Sharing data enriched by such standardized terminology can support advances in neuroscientific data exploration, from single-subject to mega-analysis. Machine readability of electrophysiological event annotations is essential for performing such analyses efficiently across software tools and packages. Hierarchical Event Descriptors (HED) provide a framework for describing events in neuroscience experiments. HED library schemas extend the standard HED schema vocabulary to include specialized vocabularies, such as standardized clinical terms for electrophysiological events. The Standardized Computer-based Organized Reporting of EEG (SCORE) defines terms for annotating EEG events, including artifacts. This study makes SCORE machine-readable by incorporating it into a HED library schema. We demonstrate the use of the HED-SCORE library schema to annotate events in example EEG data stored in Brain Imaging Data Structure (BIDS) format. Clinicians and researchers worldwide can now use the HED-SCORE library schema to annotate and then compute on electrophysiological data obtained from the human brain.Comment: 22 pages, 5 figure