THCz: Small molecules with antimicrobial activity that block cell wall lipid intermediates.

Emerging antibiotic resistance demands identification of novel antibacterial compound classes. A bacterial whole-cell screen based on pneumococcal autolysin-mediated lysis induction was developed to identify potential bacterial cell wall synthesis inhibitors. A hit class comprising a 1-amino substituted tetrahydrocarbazole (THCz) scaffold, containing two essential amine groups, displayed bactericidal activity against a broad range of gram-positive and selected gram-negative pathogens in the low micromolar range. Mode of action studies revealed that THCz inhibit cell envelope synthesis by targeting undecaprenyl pyrophosphate-containing lipid intermediates and thus simultaneously inhibit peptidoglycan, teichoic acid, and polysaccharide capsule biosynthesis. Resistance did not readily develop in vitro, and the ease of synthesizing and modifying these small molecules, as compared to natural lipid II-binding antibiotics, makes THCz promising scaffolds for development of cell wall-targeting antimicrobials.

Androgen deprivation therapy in men with prostate cancer is not associated with COVID-2019 infection.

BACKGROUND: Androgens may play a role in severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection and host responses as the virus is dependent on the androgen-regulated protein transmembrane serine protease 2 for cell entry. Studies have indicated that prostate cancer patients receiving androgen deprivation therapy (ADT) are at reduced risk of SARS-CoV-2 infection and serious complications compared with patients without ADT, but data are inconsistent. METHODS: A total of 655 prostate cancer patients who were under surveillance at two urology departments in Sweden on April 1, 2020 were included in the study as well as 240 patients with benign prostatic hyperplasia (BPH). At follow-up early in 2021, the participants completed a questionnaire containing information about symptoms compatible with coronavirus disease 2019 (COVID-19). Blood samples were also collected for the assessment of SARS-CoV-2 IgG antibodies (SARS-CoV-2 Total; Siemens). We used multivariable logistic regression models to calculate odds ratios (ORs) and 95% confidence intervals (CIs) for the association between ADT and the risk of SARS-CoV-2 infection. RESULTS: The cumulative incidence of SARS-CoV-2 seropositivity was 13.4% among patients receiving ADT and 10.4% among patients without ADT. After adjusting for potential confounders, we observed no differences in symptoms or risk of SARS-CoV-2 infection between patients with and without ADT (OR: 0.98; 95% CI: 0.52-1.85). Higher body mass index, Type 1 diabetes, and prostate cancer severity, defined by high Gleason score (8-10; OR: 2.06; 95% CI: 1.04-4.09) or elevated levels of prostate-specific antigen (>20 µg/l; OR: 2.15; 95% CI: 1.13-4.07) were associated with increased risk of SARS-CoV-2 infection. Overall, the risk of SARS-CoV-2 infection was not higher among men with prostate cancer than among men with BPH. CONCLUSIONS: Our results do not support the hypothesis that ADT use in prostate cancer patients reduces the risk or symptom severity of SARS-CoV-2 infection or that prostate cancer patients are at increased risk of COVID-19 compared with men without prostate cancer.

Managing leukemia patients via liquid biopsy and super rolling circle amplification (superRCA).

The rapidly increasing availability of sequence information for tumor patients, combined with expanding treatment options, motivates efforts to monitor the course of disease for individual patients by analyzing patient-specific mutations in liquid biopsies, as highly specific markers of the malignancy. We discuss the suitability of established molecular methods to monitor patients with malignancies, in particular leukemias, comparing these to the recently developed super rolling circle amplification technique for highly sensitive, parallel measurements of mutant sequences using readily available instruments. The very high sensitivity for tumor-specific mutations-in combination with low cost and ready access at clinics-promises to allow routine monitoring of increasing numbers of tumor patients, in order to initiate improved treatments at the earliest timepoint possible, when necessary. A method with high-enough accuracy to enable monitoring in peripheral blood rather than bone marrow samples would present a great practical advantage, not least from the patient perspective. We describe scenarios in which sufficiently sensitive, inexpensive methods for mutational analysis can provide valuable guidance for the clinician in choosing among therapeutic options and adjusting ongoing treatment and help to promptly identify recurrences of disease in treated patients.

Characterization of therapy-related acute myeloid leukemia: increasing incidence and prognostic implications.

Studies of therapy-related AML (t-AML) are usually performed in selected cohorts and reliable incidence rates are lacking. In this study, we characterized, defined the incidence over time and studied prognostic implications in all t-AML patients diagnosed in Sweden between 1997 and 2015. Data were retrieved from nationwide population-based registries. In total, 6,779 AML patients were included in the study, of whom 686 (10%) had t-AML. The median age for t-AML was 71 years and 392 (57%) patients were females. During the study period, the incidence of t-AML almost doubled with a yearly increase in t-AML of 4.5% (95% confidence interval: 2.8%-6.2%), which contributed significantly to the general increase in AML incidence over the study period. t-AML solidly constituted over 10% of all AML cases during the later period of the study. Primary diagnoses with the largest increase in incidence and decrease in mortality rate during the study period (i.e., breast and prostate cancer) contributed significantly to the increased incidence of t-AML. In multivariable analysis, t-AML was associated with poorer outcome in cytogenetically intermediate- and adverse-risk cases but t-AML had no significant impact on outcome in favorable-risk AML, including core binding leukemias, acute promyelocytic leukemia and AML with mutated NPM1 without FLT3-ITD. We conclude that there is a strong increase in incidence in t-AML over time and that t-AML constitutes a successively larger proportion of the AML cases. Furthermore, we conclude that t-AML confers a poor prognosis in cytogenetically intermediate- and adverse-risk, but not in favorable-risk AML.

Drug information sources in professional work-a questionnaire study on physicians' usage and preferences (the drug information study).

PURPOSE: This study aimed to explore physicians' use of drug information in professional work, with special focus on those working in primary care, and also in relation to personal characteristics of physicians. METHODS: A web-based questionnaire was distributed by e-mail to physicians in five regions in Sweden. The questions concerned drug-related queries at issue when searching for information, sources used, and factors of importance for the choice of source, as well as responder characteristics. RESULTS: A total of 3254 (85%) out of 3814 responding physicians stated that they searched for drug information every week. For physicians working in primary health care, the corresponding number was 585 (96%). The most common drug-related issues searched for by 76% of physicians every week concerned pharmacotherapeutic aspects (e.g., dosing), followed by adverse drug reactions (63%). For 3349 (88%) physicians, credibility was the most important factor for the choice of sources of drug information, followed by easy access online (n = 3127, 82%). Further analyses among physicians in primary care showed that some personal characteristics, like seniority, sex, and country of education, as well as research experience, were associated with usage and preferences of drug information sources. CONCLUSIONS: This study confirms that physicians often use drug information sources in professional work, in particular those who work in primary health care. Credibility and easy access are key factors for usage. Among physicians in primary care, personal factors influenced the choice of drug information sources.

Efficacy of access flap and pocket elimination procedures in the management of peri-implantitis: A systematic review and meta-analysis.

AIM: To evaluate the efficacy of access flap and pocket elimination procedures in the surgical treatment of peri-implantitis. MATERIALS AND METHODS: Systematic electronic searches (Central/MEDLINE/EMBASE) up to March 2022 were conducted to identify prospective clinical studies evaluating surgical therapy (access flap or pocket elimination procedures) of peri-implantitis. Primary outcome measures were reduction of probing depth (PD) and bleeding on probing (BOP). Risk of bias was evaluated according to study design. Meta-analysis and meta-regression were performed. Results were expressed as standardized mean effect with 95% confidence interval (CI). RESULTS: Evidence from studies directly comparing surgical with non-surgical therapy is lacking. Based on pre-post data originating from 13 prospective patient cohorts, pronounced reductions of PD (standardized mean effect: 2.2 mm; 95% CI 1.8-2.7) and BOP% (27.0; 95% CI 19.8-34.2) as well as marginal bone level gain (0.2 mm; 95% CI -0.0 to 0.5) were observed at evaluation time points ranging from 1 to 5 years. Wide prediction intervals suggested a high degree of heterogeneity. Reduction of mean PD increased by 0.7 mm (95% CI 0.5-0.9) for every millimetre in increase of mean PD at baseline. During the follow-up period ranging from 1 to 5 years, disease recurrence occurred frequently and implant loss was not uncommon. CONCLUSIONS: Access flap and pocket elimination surgery are effective procedures in the management of peri-implantitis, although rates of disease recurrence during 5 years were high. Treatment outcomes were affected by baseline conditions.

Furcation involvement and tooth loss: A registry-based retrospective cohort study.

AIM: This registry-based retrospective cohort study aimed to evaluate the impact of furcation status on the risk for molar loss. MATERIALS AND METHODS: Subjects with and without furcation involvement (FI) in 2010/2011 were identified in a nationwide registry in Sweden (age- and gender-matched sample: 381,450 subjects; 2,374,883 molars). Data on dental and periodontal status were extracted for the subsequent 10-year period. Impact of FI (at baseline or detected during follow-up) on molar loss (i.e., tooth extraction) was evaluated through multilevel logistic regression and survival analyses. RESULTS: FI had a significant impact on molar loss. FI degrees 2 and 3 resulted in adjusted risk ratios of 1.67 (95% confidence interval [CI] 1.63-1.71) and 3.30 (95% CI 3.18-3.43), respectively. Following the first detection of deep FI (degrees 2-3), estimated survival decreased by 4% at 5 years and 8% at 10 years. In addition to FI, endodontic status and probing depth were relevant risk factors for molar loss. CONCLUSIONS: Furcation status had a clinically relevant impact on the risk for molar loss. Following first detection of deep FI, however, the decline in molar survival was minor.

Prevention and management of peri-implant mucositis and peri-implantitis: A systematic review of outcome measures used in clinical studies in the last 10 years.

OBJECTIVES: To evaluate outcome measures, methods of assessment, and analysis in clinical studies on the prevention and management of peri-implant mucositis and peri-implantitis. METHODS: Systematic electronic searches (CENTRAL/MEDLINE/SCOPUS) up to April 2021 were conducted to identify longitudinal clinical studies with ≥10 patients on either the prevention or management of peri-implant diseases. Outcome measures of this analysis were the choice of outcome measures, methods of assessment, and analytical methods. Risk of bias was evaluated according to study design. Data were extracted into evidence tables and outcomes were analysed in a descriptive manner. RESULTS: The analysis of the 159 selected studies revealed that probing pocket depth (PPD) and bleeding/suppuration on probing (BOP) were reported in 89% and 87% of all studies, respectively. Additional outcome measures included plaque scores (reported in 64% of studies), radiographic outcomes (49%), soft tissue dimensions (34%), and composite outcomes (26%). Adverse events (8%) and patient-reported outcomes (6%) were only rarely mentioned. A primary outcome measure was clearly defined only in 36% of studies. Data on PPD, radiographic outcomes, and soft tissue dimensions were primarily reported as mean values and rarely as frequency distributions. For radiographic outcomes and soft tissue dimensions, it was frequently unclear how clustered data were handled. CONCLUSIONS: PPD and BOP were routinely reported in studies on the prevention and management of peri-implant mucositis and peri-implantitis, while composite outcomes, adverse events, and patient-reported outcomes were only infrequently described.

Factors influencing outcomes of surgical therapy of peri-implantitis: A secondary analysis of 1-year results from a randomized clinical study.

AIM: To identify predictors of treatment outcomes following surgical therapy of peri-implantitis. MATERIALS AND METHODS: We performed a secondary analysis of data from a randomized controlled trial (RCT) comparing access flap with or without bone replacement graft. Outcomes at 12 months were probing pocket depth (PPD), bleeding on probing (BOP), soft-tissue recession (REC) and marginal bone level (MBL) change. Multilevel regression analyses were used to identify predictors. We also built an explanatory model for residual signs of inflammation. RESULTS: Baseline PPD was the most relevant predictor, showing positive associations with final PPD, REC and MBL gain, and negative association with probability of pocket closure. Smokers presented higher residual PPD. Absence of keratinized mucosa at baseline increased the probability of BOP but was otherwise not indicative of outcomes. Plaque at 6 weeks was detrimental in terms of residual PPD and BOP. Treatment allocation had an effect on REC. Final BOP was explained by residual PPD ≥6 mm and plaque at more than two sites. CONCLUSIONS: Baseline PPD was the most relevant predictor of the outcomes of surgical therapy of peri-implantitis. Pocket closure should be a primary goal of treatment. Bone replacement grafts may be indicated in aesthetically demanding cases to reduce soft-tissue recession. The importance of smoking cessation and patient-performed plaque control is also underlined.

Prevention and management of peri-implant mucositis and peri-implantitis: A systematic review of outcome measures used in clinical studies in the last 10 years.

OBJECTIVES: To evaluate outcome measures, methods of assessment, and analysis in clinical studies on the prevention and management of peri-implant mucositis and peri-implantitis. METHODS: Systematic electronic searches (CENTRAL/MEDLINE/SCOPUS) up to April 2021 were conducted to identify longitudinal clinical studies with ≥10 patients on either the prevention or management of peri-implant diseases. Outcome measures of this analysis were the choice of outcome measures, methods of assessment, and analytical methods. Risk of bias was evaluated according to study design. Data were extracted into evidence tables and outcomes were analysed in a descriptive manner. RESULTS: The analysis of the 159 selected studies revealed that probing pocket depth (PPD) and bleeding/suppuration on probing (BOP) were reported in 89% and 87% of all studies, respectively. Additional outcome measures included plaque scores (reported in 64% of studies), radiographic outcomes (49%), soft tissue dimensions (34%), and composite outcomes (26%). Adverse events (8%) and patient-reported outcomes (6%) were only rarely mentioned. A primary outcome measure was clearly defined only in 36% of studies. Data on PPD, radiographic outcomes, and soft tissue dimensions were primarily reported as mean values and rarely as frequency distributions. For radiographic outcomes and soft tissue dimensions, it was frequently unclear how clustered data were handled. CONCLUSIONS: PPD and BOP were routinely reported in studies on the prevention and management of peri-implant mucositis and peri-implantitis, while composite outcomes, adverse events, and patient-reported outcomes were only infrequently described.

Scientific rationale for study: In 2012, recommendations on study design, key outcome measures, and reporting in clinical studies on the prevention and management of peri-implant diseases were presented. We aimed to evaluate how these recommendations were adapted and utilized in relevant studies published during the last decade. Principal findings: Recommendations on outcome measures and reporting in clinical studies on the prevention and management of peri-implant mucositis and peri-implantitis were only partially followed. Practical implications: When evaluating the evidence on the prevention and management of peri-implant diseases, the clinician should be aware of the limitations in terms of choice of outcome measures and data reporting.

Onset of periodontitis - a registry-based cohort study.

OBJECTIVES: The present retrospective registry-based cohort study aimed to identify parameters associated with the onset of periodontitis in young adults. MATERIAL AND METHODS: A total of 345 Swedish subjects were clinically examined at age 19 years (as part of an epidemiological survey) and then followed up to 31 years through the Swedish Quality Registry for Caries and Periodontal diseases (SKaPa). The registry data including periodontal parameters were obtained for the period 2010-2018 (23-31 years). Logistic regression and survival models were used to identify risk factors for periodontitis (PPD ≥6 mm at ≥2 teeth). RESULTS: The incidence of periodontitis during the 12-year observation period was 9.8%. Cigarette smoking (modified pack-years; HR 2.35, 95%CI 1.34-4.13) and increased probing pocket depth (number of sites with PPD 4-5 mm; HR 1.04, 95%CI 1.01-1.07) at 19 years were risk factors for periodontitis in subsequent young adulthood. No statistically significant association was identified for gender, snuff use, plaque and marginal bleeding scores. CONCLUSION: Cigarette smoking and increased probing pocket depth (≥4 mm) in late adolescence (19 years) were relevant risk factors for periodontitis in young adulthood. CLINICAL RELEVANCE: Our study identified cigarette smoking and increased probing depth in late adolescence as relevant risk factors of periodontitis in young adulthood. Preventive programs should therefore consider both cigarette smoking and probing pocket depths in their risk assessment.

Quantitative proteomics reveals protein dysregulation during T cell activation in multiple sclerosis patients compared to healthy controls.

BACKGROUND: Multiple sclerosis (MS) is an autoimmune, neurodegenerative disorder with a strong genetic component that acts in a complex interaction with environmental factors for disease development. CD4+ T cells are pivotal players in MS pathogenesis, where peripherally activated T cells migrate to the central nervous system leading to demyelination and axonal degeneration. Through a proteomic approach, we aim at identifying dysregulated pathways in activated T cells from MS patients as compared to healthy controls. METHODS: CD4+ T cells were purified from peripheral blood from MS patients and healthy controls by magnetic separation. Cells were left unstimulated or stimulated in vitro through the TCR and costimulatory CD28 receptor for 24 h prior to sampling. Electrospray liquid chromatography-tandem mass spectrometry was used to measure protein abundances. RESULTS: Upon T cell activation the abundance of 1801 proteins was changed. Among these proteins, we observed an enrichment of proteins expressed by MS-susceptibility genes. When comparing protein abundances in T cell samples from healthy controls and MS patients, 18 and 33 proteins were differentially expressed in unstimulated and stimulated CD4+ T cells, respectively. Moreover, 353 and 304 proteins were identified as proteins exclusively induced upon T cell activation in healthy controls and MS patients, respectively and dysregulation of the Nur77 pathway was observed only in samples from MS patients. CONCLUSIONS: Our study highlights the importance of CD4+ T cell activation for MS, as proteins that change in abundance upon T cell activation are enriched for proteins encoded by MS susceptibility genes. The results provide evidence for proteomic disturbances in T cell activation in MS, and pinpoint to dysregulation of the Nur77 pathway, a biological pathway known to limit aberrant effector T cell responses.

A risk score based on real-world data to predict early death in acute promyelocytic leukemia.

With increasingly effective treatments, early death (ED) has become the predominant reason for therapeutic failure in patients with acute promyelocytic leukemia (APL). To better prevent ED, patients with high-risk of ED must be identified. Our aim was to develop a score that predicts the risk of ED in a real-life setting. We used APL patients in the populationbased Swedish AML Registry (n=301) and a Portuguese hospital-based registry (n=129) as training and validation cohorts, respectively. The cohorts were comparable with respect to age (median, 54 and 53 years) and ED rate (19.6% and 18.6%). The score was developed by logistic regression analyses, risk-per-quantile assessment and scoring based on ridge regression coefficients from multivariable penalized logistic regression analysis. White blood cell count, platelet count and age were selected by this approach as the most significant variables for predicting ED. The score identified low-, high- and very high-risk patients with ED risks of 4.8%, 20.2% and 50.9% respectively in the training cohort and with 6.7%, 25.0% and 36.0% as corresponding values for the validation cohort. The score identified an increased risk of ED already at sub-normal and normal white blood cell counts and, consequently, it was better at predicting ED risk than the Sanz score (AUROC 0.77 vs. 0.64). In summary, we here present an externally validated and population-based risk score to predict ED risk in a real-world setting, identifying patients with the most urgent need of aggressive ED prevention. The results also suggest that increased vigilance for ED is already necessary at sub-normal/normal white blood cell counts.

Systematic review on the association between genetic polymorphisms and dental implant-related biological complications.

OBJECTIVES: The aim of this systematic review was to evaluate the association between specific genetic polymorphisms and dental implant-related biological complications in patients having a follow-up period of at least 12-months post-loading. MATERIAL AND METHODS: A sensitive search strategy was developed to identify implant-related genetic-association studies. This was performed by searching five databases. A three-stage screening (titles, abstract, full text) was carried out in duplicate and independently by two reviewers. Assessment was carried out according to the suggested scale for quality assessment of periodontal genetic-association studies and adapted to genetic analyses of implant-related studies leading to an overall final score 0-20 based on the summation of positive answers. RESULTS: The initial search resulted in 1838 articles. Sixty-seven full-text articles were assessed for eligibility and four studies met the defined inclusion criteria. IL-6 G174C, TNF-α -308, IL-1A-889 and IL-1B+3954 and CD14-159 C/T polymorphisms were evaluated. The quality assessment scores ranged from 6 to 11 positive answers from out of a maximum score of 20. The great heterogeneity among the studies did not allow a meta-analysis. CONCLUSIONS: The published evidence on genetic predisposition and implant biologic complications is limited. The small number of identified studies evaluating the association between genetic polymorphisms and peri-implant disease presented methodological and reporting inadequacies. Thus, the potential link between genetic polymorphisms and biological complications should be further investigated and clarified through well-designed clinical studies on adequately powered and appropriately included study populations.

A comparative study on periodontitis and periodontitis-associated bacteria in Somali and non-Somali children and adolescents living in Trollhättan, Sweden.

The reported prevalence of periodontitis in children and adolescents varies considerably between populations globally. This cross-sectional study compares clinical and microbiological findings on 83 Somali immigrants and 96 non-Somali children aged 10-17 years old living in Trollhättan, Sweden. The clinical examination included registration of bleeding on probing, plaque, and calculus on incisors and first molars. The distance between cemento-enamel junction and bone level was measured on bitewing radiographs. Pooled microbiological samples (1 µL) were taken from the mesial surface of 16, 11, 31, 36, and analyzed by culture and real-time polymerase chain reaction for seven periodontal associated bacterial species. The Somali participants had poorer oral hygiene and more bleeding, plaque, and calculus. Ten of the Somali but none of the non-Somali participants showed periodontal breakdown (radiographical bone loss > 3 mm), corresponding to a prevalence of 12% (95% CI: 5.9, 21.0%). The presence of A. actinomycetemcomitans was almost exclusively associated with Somali participants. Further, the JP2 clone was found in five Somalis (including two periodontitis cases) confirming the association of this clone with African populations. The Somali group showed significantly higher frequencies and numbers of Porphyromonas gingivalis and Treponema denticola, implying a mature and adult type of subgingival microbiota.

Health-related quality of life in refugee minors from Syria, Iraq and Afghanistan resettled in Sweden: a nation-wide, cross-sectional study.

PURPOSE: To examine health-related quality of life (HRQoL) in refugee minors resettled in Sweden and compare results to a European reference population, while exploring associations between sociodemographic factors and HRQoL dimensions. METHODS: A cross-sectional, nation-wide study was conducted with a stratified sample of refugee minors ages 12-15 and 16-18 from Afghanistan, Iraq and Syria, resettled in Sweden between 2014 and 2018. HRQoL was measured using KIDSCREEN-27. HRQoL dimension scores of the sample were compared to mean scores of European age and gender-matched reference population. Associations between sociodemographic factors and HRQoL dimensions were investigated with independent t tests and ANOVA. A multivariable regression analysis was performed to identify the sociodemographic factors associated with HRQoL. RESULTS: The questionnaire was sent to 10,000 potential respondents. The response rate was 26%, yielding n = 2559 refugee minors (boys 55%, girls 45%) in the study sample. Compared to European references, minors in the present study had significantly lower scores of HRQoL within psychological wellbeing and peers and social support, whereas levels for autonomy and parent/guardian relations and school environment were higher. Several sociodemographic factors were significantly associated with all HRQoL dimensions, with those 16-18 years old, having average or poor family economy, and living with an unrelated adult or family reporting lower levels of HRQoL. Minors from Afghanistan had significantly lower scores of HRQoL for all dimensions compared to those from Iraq and Syria. CONCLUSION: Refugee minors had significantly lower levels of HRQoL for psychological wellbeing and peers and social support compared to European references. Future research should further investigate this potential HRQoL gap further.

Exploring the role of the multiple sclerosis susceptibility gene CLEC16A in T cells.

C-type lectin-like domain family 16 member A (CLEC16A) is associated with autoimmune disorders, including multiple sclerosis (MS), but its functional relevance is not completely understood. CLEC16A is expressed in several immune cells, where it affects autophagic processes and receptor expression. Recently, we reported that the risk genotype of an MS-associated single nucleotide polymorphism in CLEC16A intron 19 is associated with higher expression of CLEC16A in CD4+ T cells. Here, we show that CLEC16A expression is induced in CD4+ T cells upon T cell activation. By the use of imaging flow cytometry and confocal microscopy, we demonstrate that CLEC16A is located in Rab4a-positive recycling endosomes in Jurkat TAg T cells. CLEC16A knock-down in Jurkat cells resulted in lower cell surface expression of the T cell receptor, however, this did not have a major impact on T cell activation response in vitro in Jurkat nor in human, primary CD4+ T cells.

Genomic signature of parity in the breast of premenopausal women.

BACKGROUND: Full-term pregnancy (FTP) at an early age confers long-term protection against breast cancer. Previously, we reported that a FTP imprints a specific gene expression profile in the breast of postmenopausal women. Herein, we evaluated gene expression changes induced by parity in the breast of premenopausal women. METHODS: Gene expression profiling of normal breast tissue from 30 nulliparous (NP) and 79 parous (P) premenopausal volunteers was performed using Affymetrix microarrays. In addition to a discovery/validation analysis, we conducted an analysis of gene expression differences in P vs. NP women as a function of time since last FTP. Finally, a laser capture microdissection substudy was performed to compare the gene expression profile in the whole breast biopsy with that in the epithelial and stromal tissues. RESULTS: Discovery/validation analysis identified 43 differentially expressed genes in P vs. NP breast. Analysis of expression as a function of time since FTP revealed 286 differentially expressed genes (238 up- and 48 downregulated) comparing all P vs. all NP, and/or P women whose last FTP was less than 5 years before biopsy vs. all NP women. The upregulated genes showed three expression patterns: (1) transient: genes upregulated after FTP but whose expression levels returned to NP levels. These genes were mainly related to immune response, specifically activation of T cells. (2) Long-term changing: genes upregulated following FTP, whose expression levels decreased with increasing time since FTP but did not return to NP levels. These were related to immune response and development. (3) Long-term constant: genes that remained upregulated in parous compared to nulliparous breast, independently of time since FTP. These were mainly involved in development/cell differentiation processes, and also chromatin remodeling. Lastly, we found that the gene expression in whole tissue was a weighted average of the expression in epithelial and stromal tissues. CONCLUSIONS: Genes transiently activated by FTP may have a role in protecting the mammary gland against neoplastically transformed cells through activation of T cells. Furthermore, chromatin remodeling and cell differentiation, represented by the genes that are maintained upregulated long after the FTP, may be responsible for the lasting preventive effect against breast cancer.

Quantitative proteomic analyses of CD4+ and CD8+ T cells reveal differentially expressed proteins in multiple sclerosis patients and healthy controls.

BACKGROUND: Multiple sclerosis (MS) is an autoimmune, neuroinflammatory disease, with an unclear etiology. However, T cells play a central role in the pathogenesis by crossing the blood-brain-barrier, leading to inflammation of the central nervous system and demyelination of the protective sheath surrounding the nerve fibers. MS has a complex inheritance pattern, and several studies indicate that gene interactions with environmental factors contribute to disease onset. METHODS: In the current study, we evaluated T cell dysregulation at the protein level using electrospray liquid chromatography-tandem mass spectrometry to get novel insights into immune-cell processes in MS. We have analyzed the proteomic profiles of CD4+ and CD8+ T cells purified from whole blood from 13 newly diagnosed, treatment-naive female patients with relapsing-remitting MS and 14 age- and sex-matched healthy controls. RESULTS: An overall higher protein abundance was observed in both CD4+ and CD8+ T cells from MS patients when compared to healthy controls. The differentially expressed proteins were enriched for T-cell specific activation pathways, especially CTLA4 and CD28 signaling in CD4+ T cells. When selectively analyzing proteins expressed from the genes most proximal to > 200 non-HLA MS susceptibility polymorphisms, we observed differential expression of eight proteins in T cells between MS patients and healthy controls, and there was a correlation between the genotype at three MS genetic risk loci and protein expressed from proximal genes. CONCLUSION: Our study provides evidence for proteomic differences in T cells from relapsing-remitting MS patients compared to healthy controls and also identifies dysregulation of proteins encoded from MS susceptibility genes.

Developing confidence in basic prescribing skills during medical school: a longitudinal questionnaire study investigating the effects of a modified clinical pharmacology course.

PURPOSE: To investigate if increased focus on pharmacotherapy during medical school can increase students' confidence in basic prescribing skills, that is, performing medication reviews and writing medication discharge summaries. METHODS: In 2016, the clinical pharmacology course in medical school in Gothenburg, Sweden, was modified to facilitate the students' acquisition of prescribing skills, with (i) clarified learning outcomes; (ii) supply of a list of common drugs for self-completion; (iii) instructions to practice medication reviews/discharge summaries during the ward-based education; and (iv) a concluding compulsory seminar where the students were to present prescribing-related experiences from their ward-based attendance. Questionnaires were administered to students participating in the course before (2016; n = 101) and after (2017; n = 137) implementation of the modifications. Students were asked to grade their agreement from 1 (totally disagree) to 5 (totally agree) on statements related to their perceived confidence in basic prescribing skills. RESULTS: In all, 195 students returned the questionnaire (response rate 82%; median age 24 years; 68% female). Confidence was rated higher after the modifications were implemented, both regarding medication reviews and medication discharge summaries, after vs. before 3.6 ± 1.2 vs. 3.2 ± 1.0 (P = 0.024), and 4.3 ± 0.9 vs. 3.9 ± 1.1 (P = 0.008), respectively. The adjusted odds for being confident in performing these tasks were 1.49/1.36 times greater after the course modifications (P = 0.047/0.019). Perceived confidence in performing medication reviews/summary reports was positively correlated with numbers performed (P < 0.0001). CONCLUSIONS: Modifications of the clinical pharmacology course during medical school, focusing on students' training in pharmacotherapy, was associated with increased confidence of this core skill for a physician.