Cystic fibrosis sputum DNA has NETosis characteristics and neutrophil extracellular trap release is regulated by macrophage migration-inhibitory factor

Neutrophils are the main proinflammatory cell type in chronically infected lungs of cystic fibrosis (CF) patients; however, they fail to effectively clear the colonizing pathogens. Here, we investigated the molecular composition of non-mucoid and mucoid Pseudomonas aeruginosa-induced neutrophil extracellular traps (NETs) in vitro and compared them to the DNA-protein complexes present in the CF sputum. The protein composition of P. aeruginosa-induced NET fragments revealed that irrespective of the inducing stimuli, NET fragments were decorated with a conserved set of proteins. The DNA-protein complexes derived from CF sputum were consistent with NETosis and shared a similar protein signature, suggesting that the majority of the extracellular DNA was NET derived. The ability of polymorphonuclear leukocytes to produce NETs in response to P. aeruginosa was driven by macrophage migration-inhibitory factor (MIF) by promoting mitogen-activated protein kinase. Analysis of 132 CF patient samples revealed that elevated MIF protein levels correlated with poorer lung function. We suggest that targeting MIF by small molecular inhibitors might reduce the presence of extracellular DNA and serve as an adjunct to the use of antimicrobial drugs that could ultimately reduce bacterial fitness in the lungs during the later stages of CF disease

Reduced risk of myocardial infarct and revascularization following coronary artery bypass grafting compared with percutaneous coronary intervention in patients with chronic kidney disease

Coronary atherosclerotic disease is highly prevalent in chronic kidney disease (CKD). Although revascularization improves outcomes, procedural risks are increased in CKD and unbiased data comparing bypass surgery (CABG) and percutaneous intervention (PCI) in CKD are sparse. To compare outcomes of CABG and PCI in stage 3-5 CKD, we identified randomized trials comparing these procedures. Investigators were contacted to obtain individual, patient-level data. Ten of 27 trials meeting inclusion criteria provided data. These trials enrolled 3993 patients encompassing 526 patients with stage 3-5 CKD of which 137 were stage 3b-5 CKD. Among individuals with stage 3-5 CKD survival through 5-years was not different following CABG compared with PCI (hazard ratio 0.99, 95% confidence interval: 0.67, 1.46) or stage 3b-5 CKD (1.29: 0.68, 2.46). However, CKD modified the impact on survival free from myocardial infarction: it was not different between CABG and PCI for individuals with preserved kidney function (0.97: 0.80, 1.17), but was significantly lower following CABG in stage 3-5 CKD (0.49: 0.29, 0.82) and stage 3b-5 CKD (0.23: 0.09, 0.58). Repeat revascularization was reduced following CABG compared with PCI regardless of baseline kidney function. Results were limited by unavailability of data from several trials and paucity of enrolled patients with stage 4-5 CKD. Thus, our patient-level meta-analysis of individuals with CKD randomized to CABG versus PCI suggests that CABG significantly reduces the risk of subsequent myocardial infarction and revascularization without impacting survival in these patients

Labeling of alprenolol with fluorescent aryl iodide as a reagent based on Mizoroki-Heck coupling reaction.

A novel fluorescent labeling method for alprenolol was developed based on Mizoroki-Heck coupling reaction. We designed and synthesized fluorescent aryl iodide, 4-(4,5-diphenyl-1H-imidazol-2-yl)iodobenzene (DIBI) as a labeling reagent. DIBI has a lophine skeleton carrying an iodide atom acting as fluorophore and reactive center, respectively. In order to evaluate the usefulness of DIBI, a high-performance liquid chromatography (HPLC) with fluorescence detection method was developed for the determination of alprenolol as a model compound of terminal double bond. The fluorescent labeling of alprenolol with DIBI was achieved in the presence of palladium acetate as a catalyst, and the labeled alprenolol was detected fluorometrically. In addition, it was found that the fluorescence of DIBI derivative increased and red shifted when compared with that of DIBI. Furthermore, the proposed method could be applied to determine the alprenolol concentration in rat plasma after administration of alprenolol without interferences from biological components. The detection limit (S/N=3) for alprenolol in rat plasma was 0.74 ng/mL (30 fmol on column)

Updating the approaches to define susceptibility and resistance to anti-tuberculosis agents: implications for diagnosis and treatment

11 páginas, 2 figuras, 1 tablaInappropriately high breakpoints have resulted in systematic false-susceptible AST results to anti-TB drugs. MIC, PK/PD and clinical outcome data should be combined when setting breakpoints to minimise the emergence and spread of antimicrobial resistance.I. Comas was supported by PID2019-104477RB-I00 from the Spanish Science Ministry and by ERC (CoG 101001038)Peer reviewe

31st Annual Meeting and Associated Programs of the Society for Immunotherapy of Cancer (SITC 2016) : part two

Background The immunological escape of tumors represents one of the main ob- stacles to the treatment of malignancies. The blockade of PD-1 or CTLA-4 receptors represented a milestone in the history of immunotherapy. However, immune checkpoint inhibitors seem to be effective in specific cohorts of patients. It has been proposed that their efficacy relies on the presence of an immunological response. Thus, we hypothesized that disruption of the PD-L1/PD-1 axis would synergize with our oncolytic vaccine platform PeptiCRAd. Methods We used murine B16OVA in vivo tumor models and flow cytometry analysis to investigate the immunological background. Results First, we found that high-burden B16OVA tumors were refractory to combination immunotherapy. However, with a more aggressive schedule, tumors with a lower burden were more susceptible to the combination of PeptiCRAd and PD-L1 blockade. The therapy signifi- cantly increased the median survival of mice (Fig. 7). Interestingly, the reduced growth of contralaterally injected B16F10 cells sug- gested the presence of a long lasting immunological memory also against non-targeted antigens. Concerning the functional state of tumor infiltrating lymphocytes (TILs), we found that all the immune therapies would enhance the percentage of activated (PD-1pos TIM- 3neg) T lymphocytes and reduce the amount of exhausted (PD-1pos TIM-3pos) cells compared to placebo. As expected, we found that PeptiCRAd monotherapy could increase the number of antigen spe- cific CD8+ T cells compared to other treatments. However, only the combination with PD-L1 blockade could significantly increase the ra- tio between activated and exhausted pentamer positive cells (p= 0.0058), suggesting that by disrupting the PD-1/PD-L1 axis we could decrease the amount of dysfunctional antigen specific T cells. We ob- served that the anatomical location deeply influenced the state of CD4+ and CD8+ T lymphocytes. In fact, TIM-3 expression was in- creased by 2 fold on TILs compared to splenic and lymphoid T cells. In the CD8+ compartment, the expression of PD-1 on the surface seemed to be restricted to the tumor micro-environment, while CD4 + T cells had a high expression of PD-1 also in lymphoid organs. Interestingly, we found that the levels of PD-1 were significantly higher on CD8+ T cells than on CD4+ T cells into the tumor micro- environment (p < 0.0001). Conclusions In conclusion, we demonstrated that the efficacy of immune check- point inhibitors might be strongly enhanced by their combination with cancer vaccines. PeptiCRAd was able to increase the number of antigen-specific T cells and PD-L1 blockade prevented their exhaus- tion, resulting in long-lasting immunological memory and increased median survival

Medical student simulation training in intrauterine contraception insertion and removal: an intervention to improve comfort, skill, and attitudes

Background: Opportunities for medical students to place intrauterine contraception (IUC) in patients are rare. Our objective was to determine whether participation in an IUC insertion and removal simulation exercise would increase medical students’ comfort level with, attitudes towards, and willingness to recommend IUC. Methods: A prospective cohort study was undertaken in all students completing the obstetrics and gynecology clerkship at a major academic hospital during the 2010–2011 academic year. The exposure consisted of a 45-minute interactive didactic session and a 30-minute, hands-on practicum in IUC placement and removal using medical instruments and realistic pelvic models. Both levonorgestrel and Cu380A IUC devices were utilized. Participants completed a pre- and post-simulation survey instrument, designed to examine students’ IUC-specific knowledge, comfort, and attitudes. Pre- and post-simulation responses were compared by McNemar’s test for paired samples. Results: Thirty-five paired pre- and post-simulation surveys were analyzed, representing a 78 % response rate. Composite IUC-related knowledge scores increased by a median of 3 out of 10 points after the intervention (p < 0.01). Students were significantly more comfortable counseling patients about IUC as well as inserting IUC after the intervention, compared to before. Seven (20 %) students before, compared to 27 (77 %) after, agreed with the statement, “I feel comfortable placing an IUC in a patient under the supervision of an experienced doctor” (p < 0.01). Students developed significantly more favorable attitudes towards IUC through the intervention. Nineteen (54 %) participants before, compared to 27 (77 %) after, agreed with the statement, “I would recommend an IUC to my family member” (p = 0.02). Conclusions: A hands-on simulation during the obstetrics and gynecology clerkship increased medical students’ knowledge of and comfort with IUC and resulted in more favorable attitudes toward the method. Intrauterine contraception simulation in medical curricula may help expand utilization of this effective contraceptive method

Relationship Of Job Role And Clinic Type To Perceived Stigma And Occupational Stress Among Abortion Workers

Objective: To describe the role of worksite characteristics, job role and abortion stigma in clinical abortion workers’ occupational stress. Study design: We recruited nurses, medical assistants and counseling staff who provide abortion care at five hospitals and four freestanding clinics in Massachusetts. Study instruments assessed individual-level abortion stigma (the perception that others treat abortion as shameful, dirty and socially taboo) using the Abortion Providers Stigma Scale; job strain through the Job Content Questionnaire; and emotional burnout through the Depersonalization, Emotional Exhaustion and Personal Accomplishment subscales of the Maslach Burnout Inventory. Multivariable linear and logistic regression modeling tested for associations between job role, worksite type, stigma, job strain and burnout, accounting for demographic and job characteristics. Results: Of 205 eligible workers, 136 participated (66%). Forty-one percent were medical assistants, 50% nurses, and 9% counselors. In adjusted models, abortion workers with high stigma had increased odds of experiencing job strain [adjusted odds ratio (aOR) 3.94, 95% CI 1.19–13.05]. Compared to working in a freestanding clinic, working in a hospital was associated with lower odds of experiencing the low personal accomplishment dimension of burnout (aOR 0.49, 95% CI 0.29–0.84) and lower odds of experiencing the depersonalization dimension of burnout (aOR 0.05, 95% CI 0.01–0.28). Counselors had higher odds of depersonalization compared to nurses (aOR 4.56, 95% CI 2.3–9.0). Conclusion: Hospital-based abortion workers experience lower risk for burnout than comparable workers in freestanding clinics, accounting for abortion stigma and job characteristics. Implications: Implementing structured supports to alter workflows or improve coping among workers in freestanding abortion clinics may help mitigate risk of worker burnout and associated turnover

High-risk sexual behaviors while on depot medroxyprogesterone acetate as compared to oral contraception

Abstract Background Depot medroxyprogesterone acetate (DMPA) contraceptive use is associated with an increased risk for Chlamydia infection. However, prior studies inadequately account for potential differences in sexual behavior between users of DMPA and users of other contraceptive methods. In this study we compare sexual risk-taking behavior in women using DMPA to women using oral contraceptive pills (OCP) to assess risk of Chlamydia trachomatis infection. Methods In this cross-sectional study of 630 reproductive-aged women seeking routine gynecologic care (449 OCP and 181 DMPA users) sexual risk-taking was evaluated by use of the Safe Sex Behavior Questionnaire, a validated measure of sexual behaviors and attitudes. All women were screened for Chlamydia . Logistic regression estimated the association of contraceptive choice, sexual behaviors, and Chlamydia infection. Results Oral contraceptive pill users differed from DMPA users in age, race, marital status, education level, and pregnancy history ( p -values all <0.05). Oral contraceptive pill users had used their method of contraception for longer average duration ( p \u2009<\u20090.01) and reported greater frequency of condom use ( p \u2009<\u20090.01). Eleven (2.5%) OCP and 2 (1.1%) DMPA users had Chlamydia ( p \u2009=\u2009NS). Conclusions Oral contraceptive pill and DMPA users differed with respect to both demographic factors and frequency of condom use. Odds of current Chlamydia infection did not differ between OCP and DMPA users when controlling for sexual risk-taking or demographic factors, though due to low Chlamydia rates in our population, this study was underpowered to detect this difference

Pain related outcomes.

BackgroundEnhanced recovery is the gold standard in modern perioperative management, including that for cesarean deliveries. However, qualitative and quantitative data on the physical and psychological recovery of women after vaginal childbirth are limited. Whether neuraxial labor analgesia influences postpartum recovery is unknown.MethodsPrimiparous women anticipating a vaginal childbirth between January 2020 and May 2021 were enrolled. Women with major comorbidities or postpartum complications and those who underwent a cesarean delivery were excluded. Daily step count was measured using a wrist-worn activity tracker (FitbitTM Inspire HR) for 120 hours after vaginal childbirth. Subjective fatigue levels and health-related quality of life were assessed using the Multidimensional Fatigue Inventory (MFI) and EuroQol 5 Dimension 5 Level (EQ-5D-5L), respectively, at the 3rd trimester antenatal visit, on postpartum day 1 and 3, and at the one-month postpartum visit. Rest and dynamic pain scores and the location of pain were documented by participants during postpartum hospitalization.ResultsAmong 300 women who were enrolled antenatally, 95 and 116 had a vaginal delivery without (NCB group) and with (EPL group) epidural analgesia, respectively. The median number of steps per 24 hours increased daily in both groups, and no significant difference was detected between the groups. Postpartum pain was mild overall, with median rest and dynamic pain scores being less than 4 and similar between the groups. MFI and EQ-5D-5L scores were the worst on postpartum day 1 in both groups and gradually improved to antepartum level by the one-month postpartum visit. Higher MFI score on postpartum day 1, but not the use of epidural analgesia, was associated with lower odds of achieving adequate postpartum ambulation (defined as >3500 steps between 48 and 72 hours postpartum).ConclusionThe use of epidural analgesia was not associated with worse recovery outcomes during postpartum hospitalization.Trial registrationUMIN-CTR, #UMIN000039343, registered on January 31, 2020.</div