Identification of target T cell epitopes for a therapeutic HPV16 vaccine

To rationally design therapeutic human papillomavirus (HPV) vaccines, it is important to know which T cell epitopes are present on HPV-transformed cells. HPV affects the cellular antigen processing machinery, thus not every epitope derived from viral proteins is necessarily presented by human leukocyte antigen (HLA) molecules on the cancer cell surface. HPV16 has been identified as the causative agent in 50% of all cervical cancer cases and in approximately 95% of all extra-cervical mucosal HPV-induced tumors. The transforming potential of high-risk HPVs is mediated by two consistently expressed viral oncoproteins, E6 and E7. As the induction and maintenance of the malignant phenotype depend on these two proteins, they are ideal targets for immunotherapy. A therapeutic vaccine that is applicable to everyone without prior HLA typing needs to contain epitopes for the major HLA types. To date, HPV T cell epitopes have mostly been determined for the most prevalent HLA type, HLA-A2. We now aim to identify HPV16 E6 and E7 T cell epitopes for the HLA-A3, HLA-A11 and HLA-A24 supertypes. Different in silico prediction algorithms were used to predict prospective epitopes from the E6 and E7 proteins derived from the HPV16 reference sequence for the mentioned HLA supertypes. In total, 74 epitopes, comprised of 8 to 11mer peptides, were predicted for HLA-A3, 96 epitopes for HLA-A11 and 95 epitopes for HLA-A24. In competition-based cellular binding assays, 22 previously known binding peptides were confirmed and 78 novel binding peptides were identified. Additionally, HPV16 variants harbored in our HPV16-positive cell line collection were determined and peptide binding to HLA-A24 was shown for seven out of 20 tested HPV16 variant peptides. Evaluation of prediction server performance based on the generated data suggested different optimal prediction servers depending on peptide length and on the HLA type. NetMHC and NetMHCcons were shown to be the best predictors overall. Immunogenicity of identified binding peptides was investigated by screening long-term memory responses in healthy donors. To this end, peptide-specific short-term T cell lines were generated from peripheral blood mononuclear cells, which were HLA typed, stimulated with HLA-matching peptides and cultured for twelve days. Several peptides were identified to be immunogenic. Immunogenicity of the four most promising candidate peptides for HLA-A24 could be further confirmed by generation of peptide-specific long-term T cell lines from healthy HLA-A24-positive donors and subsequent coculture with autologous B cells pulsed with the respective peptide. Functional assays, such as IFNγ ELISpot assays and cytotoxicity assays, determined the best vaccine candidates. In conclusion, several novel HPV16 E6 and E7 CD8+ T cell epitopes were identified.Verified epitopes are the basis of rational therapeutic vaccine design and are also important for immunomonitoring purposes. In addition, they can be employed as a tool for the development of other immunotherapies such as adoptive T cell transfer with transgenic T cell receptors

The effect of motivational orientation on academic achievement and persistence of rural community college students: A test of a motivational model from a self-determination theory perspective

Understanding the relationship between motivation, academic achievement, and persistence may help higher education administrators develop plans for increasing student engagement and student success. Prior research on higher education indicated connections between motivation, academic achievement, and persistence, but there has been limited research that relates these concepts to the rural community college setting. The purpose of this study was to examine the motivation to pursue higher education by rural community college students as well as the relationship between motivation, academic achievement, and persistence. Furthermore, it was important to assess the way student variables may moderate the relationship between motivation, academic achievement, and persistence, including demographic characteristics such as gender, race/ethnicity, childhood residence, and socioeconomic status as well as participation characteristics such as enrollment status and Oregon Promise program participation. The study used a quantitative approach to analyze the motivating factors of pursuing higher education as well as how these factors affect the academic success of students. The dependent outcomes of the study included academic achievement (measured by grade point average) and intention to persist. Self-determination theory provided the guiding theoretical perspective for the study. Through a descriptive design, the researcher studied various extrinsic and intrinsic motivating factors that led rural community college students to pursue higher education. In addition, the researcher measured the academic achievement and the intent to persist of these students in order to consider the relationship between motivation, academic achievement, and persistence within higher education. The unit of analysis involved individual students at rural community colleges in Oregon. Data were collected using a web-based questionnaire, which was sent one time in the fall term 2017 to these students. Three community colleges in Oregon participated in the study and sent the study information, including the link to the web-based questionnaire, via email to their students. The web-based questionnaire included demographic items, student participation items, self-reported grade point average, items related to intention to persist as well as the Competence Motivation Scale, the Need for Relatedness at College Questionnaire, and the Autonomous Motivation Scale. More than 700 students responded to the survey; however, data cleaning revealed only 346 usable cases that included both of the dependent variables, grade point average and intention to persist. In order to analyze the data, the following techniques were used. Data cleaning was conducted in Microsoft Excel. Descriptive data analysis was completed in EQS – Structural Equation Modeling Software (Bentler, 2006) and IBM SPSS (2018). Exploratory factor analyses and correlational analyses were also performed on the dataset. EQS – Structural Equation Modeling Software was used to perform structural equation modeling in order to test the structural pathways between motivational orientation, academic achievement, and persistence, as well as to test for moderation effects by assessing the first of the hypothesized model for the entire sample of students as well as by the student moderating variables. The results of the data were described by the overall model and moderated models. The standardized coefficient for the path between competence motivational orientation and intention to persist as well as competence and grade point average were positive and significant, indicating that students who attended college motivated by the need for competence were more likely to report intentions to persist and were more likely to achieve higher grade point averages. On the other hand, the standardized coefficient for the path between autonomy motivational orientation and grade point average was negative and significant, indicating that students who attended college to fulfill needs for autonomy were less likely to achieve higher grade point averages. In addition, there were several significant findings from the moderated models. According to the results of the free versus constrained nested comparison, socioeconomic status (higher versus lower) and Oregon Promise participation (received grant versus did not receive grant) moderated at least one of the pathways in the overall model. Examination of the critical ratio for differences between parameters in the configural model showed significant differences in the socioeconomic status model (lower versus higher socioeconomic status) and Oregon Promise model (grant recipient versus non-recipient). The results of this study suggested that important relationships exist between college student motivational orientation and academic success. Furthermore, the study showed that self-determination theory contributes to the understanding of college student academic achievement and persistence. The structural equation modeling indicated that there were significant and positive relationships between the motivational orientation of competence and intention to persist as well as grade point average. Students who attended college to fulfill a need for competence were concerned about their achievements, knowledge, and skills. If this need for competence was fulfilled, these students may experience greater levels of intrinsic motivation, which may lead to higher academic achievement and greater likelihood for persistence. Faculty could help increase students’ perceived competence by providing positive feedback to their students. In addition, college foundations could offer scholarship awards to students who meet some academic merit criteria. Although there were limitations to the study, it still provided significant results that would be beneficial to share with community college administrators, faculty, and student services professionals, among others

Abnormal small heat shock protein interactions involving neuropathy-associated HSP22 (HSPB8) mutants

Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/154632/1/fsb2fj065911fje.pd

Three-dimensional architecture and biogenesis of membrane structures associated with hepatitis C virus replication

All positive strand RNA viruses are known to replicate their genomes in close association with intracellular membranes. In case of the hepatitis C virus (HCV), a member of the family Flaviviridae, infected cells contain accumulations of vesicles forming a membranous web (MW) that is thought to be the site of viral RNA replication. However, little is known about the biogenesis and three-dimensional structure of the MW. In this study we used a combination of immunofluorescence- and electron microscopy (EM)-based methods to analyze the membranous structures induced by HCV in infected cells. We found that the MW is derived primarily from the endoplasmic reticulum (ER) and contains markers of rough ER as well as markers of early and late endosomes, COP vesicles, mitochondria and lipid droplets (LDs). The main constituents of the MW are single and double membrane vesicles (DMVs). The latter predominate and the kinetic of their appearance correlates with kinetics of viral RNA replication. DMVs are induced primarily by NS5A whereas NS4B induces single membrane vesicles arguing that MW formation requires the concerted action of several HCV replicase proteins. Three-dimensional reconstructions identify DMVs as protrusions from the ER membrane into the cytosol, frequently connected to the ER membrane via a neck-like structure. In addition, late in infection multi-membrane vesicles become evident, presumably as a result of a stress-induced reaction. Thus, the morphology of the membranous rearrangements induced in HCV-infected cells resemble those of the unrelated picorna-, corona- and arteriviruses, but are clearly distinct from those of the closely related flaviviruses. These results reveal unexpected similarities between HCV and distantly related positive-strand RNA viruses presumably reflecting similarities in cellular pathways exploited by these viruses to establish their membranous replication factories

Bacteria producing contractile phage tail-like particles (CPTPs) are promising alternatives to conventional pesticides

Diese Übersichtsarbeit verfolgt das Ziel, über bakterielle kontraktile Phagenderivate (englisch CPTPs) und ihr Potenzial als effiziente und Schadorganismus-spezifische Alternativen zu konventionellen chemischen Pflanzenschutzmitteln in der Land-/Forstwirtschaft und im Gartenbau zu informieren. CPTPs werden von verschiedenen Bakterien in diversen Habitaten für den interbakteriellen Konkurrenzkampf sowie zur Beeinflussung eukaryotischer Wirte, wie Pilze und Insekten, verwendet. Diese Arbeit präsentiert interessante und bemerkenswerte Beispiele für den vielfältigen Einsatz von CPTPs als leistungsfähige biologische Bekämpfungsmittel. Wir stellen die verschiedenen Typen von CPTPs vor und legen einen besonderen Fokus auf diejenigen, die eine Wirkung gegen Schadinsekten besitzen. Zusätzlich stellen wir zwei kürzlich etablierte Webservices vor, die das permanent wachsende Wissen über CPTPs mit einem Auswahlverfahren für die besten Bakterienkandidaten kombinieren, um eine zielgerichtete Anwendung der CPTPs in der nachhaltigen Pflanzenproduk­tion zu ermöglichen.This mini-review aims at raising the interest in contractile phage tail-like particles (CPTPs) of bacteria as an efficient and pest-specific alternative to conventional chemical pesticides in agriculture, horticulture and forestry. CPTPs are used by various bacteria in diverse environments for interbacterial competition or for manipulation of eukaryotic hosts, such as fungi or insects. This review gives examples for the versatile use of CPTPs as powerful biological control agents. We introduce the different types of CPTPs with a special focus on those with activity against insect plant pests. In addition, we present two currently established web services that combine the permanently increasing knowledge on CPTPs with a selection approach of the best candidate bacteria for targeted CPTP application in sustainable plant production

Расчет параметров гидротранспорта высококонцентрированных гидросмесей в условиях предприятий Кривбасса

Для технології складування на підприємствах Кривбасу відходів збагачення, згущених до концентрації пасти, запропоновано методики розрахунків параметрів та режимів роботи/ гідротранспортних установок з урахуванням гідравлічних та реологічних характеристик.The methods of calculation of parameters and regimes of hydrotransport plants operation for technology of stocking of cleaning rejects condensed till paste concentration at Krivbas enterprises taking into account hydraulic and rheological characteristics are offered

Severe infections of Panton-Valentine leukocidin positive Staphylococcus aureus in children

Infections caused by Panton-Valentine leukocidin-positive Staphylococcus aureus (PVL-SA) mostly present as recurrent skin abscesses and furunculosis. However, life-threatening infections (eg, necrotizing pneumonia, necrotizing fasciitis, and osteomyelitis) caused by PVL-SA have also been reported.We assessed the clinical phenotype, frequency, clinical implications (surgery, length of treatment in hospitals/intensive care units, and antibiotic treatments), and potential preventability of severe PVL-SA infections in children.Total, 75 children treated for PVL-SA infections in our in- and outpatient units from 2012 to 2017 were included in this retrospective study.Ten out of 75 children contracted severe infections (PVL-methicillin resistant S aureus n = 4) including necrotizing pneumonia (n = 4), necrotizing fasciitis (n = 2), pyomyositis (n = 2; including 1 patient who also had pneumonia), mastoiditis with cerebellitis (n = 1), preorbital cellulitis (n = 1), and recurrent deep furunculosis in an immunosuppressed patient (n = 1). Specific complications of PVL-SA infections were venous thrombosis (n = 2), sepsis (n = 5), respiratory failure (n = 5), and acute respiratory distress syndrome (n = 3). The median duration of hospital stay was 14 days (range 5-52 days). In 6 out of 10 patients a history suggestive for PVL-SA colonization in the patient or close family members before hospital admission was identified.PVL-SA causes severe to life-threatening infections requiring lengthy treatments in hospital in a substantial percentage of symptomatic PVL-SA colonized children. More than 50% of severe infections might be prevented by prompt testing for PVL-SA in individuals with a history of abscesses or furunculosis, followed by decolonization measures

Case report: Cystic fibrosis with kwashiorkor: A rare presentation in the era of universal newborn screening

BackgroundUniversal newborn screening changed the way medical providers think about the presentation of cystic fibrosis (CF). Before implementation of universal screening, it was common for children with CF to present with failure to thrive, nutritional deficiencies, and recurrent infections. Now, nearly all cases of CF are diagnosed by newborn screening shortly after birth before significant symptoms develop. Therefore, providers often do not consider this illness in the setting of a normal newborn screen. Newborn screening significantly decreases the risk of complications in early childhood, yet definitive testing should be pursued if a patient with negative newborn screening presents with symptoms consistent with CF, including severe failure to thrive, metabolic alkalosis due to significant salt losses, or recurrent respiratory infections.Case presentationWe present a case of a 6-month-old infant male with kwashiorkor, severe edema, multiple vitamin deficiencies, hematemesis secondary to coagulopathy, and diffuse erythematous rash, all secondary to severe pancreatic insufficiency. His first newborn screen had an immunoreactive trypsinogen (IRT) value below the state cut-off value, so additional testing was not performed, and his growth trajectory appeared reassuring. He was ultimately diagnosed with CF by genetic testing and confirmatory sweat chloride testing, in the setting of his parents being known CF carriers and his severe presentation being clinically consistent with CF. Acutely, management with supplemental albumin, furosemide, potassium, and vitamin K was initiated to correct the presenting hypoalbuminemia, edema, and coagulopathy. Later, pancreatic enzyme supplementation and additional vitamins and minerals were added to manage ongoing deficiencies from pancreatic insufficiency. With appropriate treatment, his vitamin deficiencies and edema resolved, and his growth improved.ConclusionDue to universal newborn screening, symptomatic presentation of CF is rare and presentation with kwashiorkor is extremely rare in resource-rich communities. The diagnosis of CF was delayed in our patient because of a normal newborn screen and falsely reassuring growth, which after diagnosis was determined to be secondary to severe edematous malnutrition. This case highlights that newborn screening is a useful but imperfect tool. Clinicians should continue to have suspicion for CF in the right clinical context, even in the setting of normal newborn screen results

Evidence-based Review on the Use of Proton Therapy in Lymphoma From the Particle Therapy Cooperative Group (PTCOG) Lymphoma Subcommittee.

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ACR Appropriateness Criteria® Hodgkin Lymphoma-Favorable Prognosis Stage I and II

This topic addresses the treatment of newly diagnosed patients with favorable prognosis stage I and II Hodgkin lymphoma. In most cases, combined modality therapy (chemotherapy followed by involved site radiation therapy) constitutes the current standard of care. The American College of Radiology Appropriateness Criteria are evidence-based guidelines for specific clinical conditions that are reviewed annually by a multidisciplinary expert panel. The guideline development and revision include an extensive analysis of current medical literature from peer-reviewed journals and the application of well-established methodologies (RAND/UCLA Appropriateness Method and Grading of Recommendations Assessment, Development, and Evaluation or GRADE) to rate the appropriateness of imaging and treatment procedures for specific clinical scenarios. In those instances where evidence is lacking or equivocal, expert opinion may supplement the available evidence to recommend imaging or treatment. By combining the most recent medical literature and expert opinion, this revised guideline can aid clinicians in the appropriate use of combined modality therapy for favorable prognosis stage I and II Hodgkin lymphoma. Increasing information about the late effects of treatment has led to attempts to decrease toxicity by using less chemotherapy (decreased duration and/or intensity or different agents) and less radiation therapy (reduced volume and/or dose) while maintaining excellent efficacy