Characteristics of the Alternative Phenotype of Microglia/Macrophages and its Modulation in Experimental Gliomas

Microglia (brain resident macrophages) accumulate in malignant gliomas and instead of initiating the anti-tumor response, they switch to a pro-invasive phenotype, support tumor growth, invasion, angiogenesis and immunosuppression by release of cytokines/chemokines and extracellular matrix proteases. Using immunofluorescence and flow cytometry, we demonstrate an early accumulation of activated microglia followed by accumulation of macrophages in experimental murine EGFP-GL261 gliomas. Those cells acquire the alternative phenotype, as evidenced by evaluation of the production of ten pro/anti-inflammatory cytokines and expression profiling of 28 genes in magnetically-sorted CD11b+ cells from tumor tissues. Furthermore, we show that infiltration of implanted gliomas by amoeboid, Iba1-positive cells can be reduced by a systematically injected cyclosporine A (CsA) two or eight days after cell inoculation. The up-regulated levels of IL-10 and GM-CSF, increased expression of genes characteristic for the alternative and pro-invasive phenotype (arg-1, mt1-mmp, cxcl14) in glioma-derived CD11b+ cells as well as enhanced angiogenesis and tumor growth were reduced in CsA-treated mice. Our findings define for the first time kinetics and biochemical characteristics of glioma-infiltrating microglia/macrophages. Inhibition of the alternative activation of tumor-infiltrating macrophages significantly reduced tumor growth. Thus, blockade of microglia/macrophage infiltration and their pro-invasive functions could be a novel therapeutic strategy in malignant gliomas

The freshwater Sponge Ephydatia Fluviatilis harbours diverse pseudomonas species (Gammaproteobacteria, Pseudomonadales) with broad-spectrum antimicrobial activity

Bacteria are believed to play an important role in the fitness and biochemistry of sponges (Porifera). Pseudomonas species (Gammaproteobacteria, Pseudomonadales) are capable of colonizing a broad range of eukaryotic hosts, but knowledge of their diversity and function in freshwater invertebrates is rudimentary. We assessed the diversity, structure and antimicrobial activities of Pseudomonas spp. in the freshwater sponge Ephydatia fluviatilis. Polymerase Chain Reaction - Denaturing Gradient Gel Electrophoresis (PCR-DGGE) fingerprints of the global regulator gene gacA revealed distinct structures between sponge-associated and free-living Pseudomonas communities, unveiling previously unsuspected diversity of these assemblages in freshwater. Community structures varied across E. fluviatilis specimens, yet specific gacA phylotypes could be detected by PCR-DGGE in almost all sponge individuals sampled over two consecutive years. By means of whole-genome fingerprinting, 39 distinct genotypes were found within 90 fluorescent Pseudomonas isolates retrieved from E. fluviatilis. High frequency of in vitro antibacterial (49%), antiprotozoan (35%) and anti-oomycetal (32%) activities was found among these isolates, contrasting less-pronounced basidiomycetal (17%) and ascomycetal (8%) antagonism. Culture extracts of highly predation-resistant isolates rapidly caused complete immobility or lysis of cells of the protozoan Colpoda steinii. Isolates tentatively identified as P. jessenii, P. protegens and P. oryzihabitans showed conspicuous inhibitory traits and correspondence with dominant sponge-associated phylotypes registered by cultivation-independent analysis. Our findings suggest that E. fluviatilis hosts both transient and persistent Pseudomonas symbionts displaying antimicrobial activities of potential ecological and biotechnological value.European Regional Development Fund (ERDF) through the COMPETE (Operational Competitiveness Programme); national funds through FCT (Foundation for Science and Technology) [PEst-C/MAR/LA0015/2011]; FCT-funded project [PTDC/BIA-MIC/3865/2012]; Federation of European Microbiological Societies (FEMS)info:eu-repo/semantics/publishedVersio

Effects of alirocumab on types of myocardial infarction: insights from the ODYSSEY OUTCOMES trial

Aims  The third Universal Definition of Myocardial Infarction (MI) Task Force classified MIs into five types: Type 1, spontaneous; Type 2, related to oxygen supply/demand imbalance; Type 3, fatal without ascertainment of cardiac biomarkers; Type 4, related to percutaneous coronary intervention; and Type 5, related to coronary artery bypass surgery. Low-density lipoprotein cholesterol (LDL-C) reduction with statins and proprotein convertase subtilisin–kexin Type 9 (PCSK9) inhibitors reduces risk of MI, but less is known about effects on types of MI. ODYSSEY OUTCOMES compared the PCSK9 inhibitor alirocumab with placebo in 18 924 patients with recent acute coronary syndrome (ACS) and elevated LDL-C (≥1.8 mmol/L) despite intensive statin therapy. In a pre-specified analysis, we assessed the effects of alirocumab on types of MI. Methods and results  Median follow-up was 2.8 years. Myocardial infarction types were prospectively adjudicated and classified. Of 1860 total MIs, 1223 (65.8%) were adjudicated as Type 1, 386 (20.8%) as Type 2, and 244 (13.1%) as Type 4. Few events were Type 3 (n = 2) or Type 5 (n = 5). Alirocumab reduced first MIs [hazard ratio (HR) 0.85, 95% confidence interval (CI) 0.77–0.95; P = 0.003], with reductions in both Type 1 (HR 0.87, 95% CI 0.77–0.99; P = 0.032) and Type 2 (0.77, 0.61–0.97; P = 0.025), but not Type 4 MI. Conclusion  After ACS, alirocumab added to intensive statin therapy favourably impacted on Type 1 and 2 MIs. The data indicate for the first time that a lipid-lowering therapy can attenuate the risk of Type 2 MI. Low-density lipoprotein cholesterol reduction below levels achievable with statins is an effective preventive strategy for both MI types.For complete list of authors see http://dx.doi.org/10.1093/eurheartj/ehz299</p

Effect of alirocumab on mortality after acute coronary syndromes. An analysis of the ODYSSEY OUTCOMES randomized clinical trial

Background: Previous trials of PCSK9 (proprotein convertase subtilisin-kexin type 9) inhibitors demonstrated reductions in major adverse cardiovascular events, but not death. We assessed the effects of alirocumab on death after index acute coronary syndrome. Methods: ODYSSEY OUTCOMES (Evaluation of Cardiovascular Outcomes After an Acute Coronary Syndrome During Treatment With Alirocumab) was a double-blind, randomized comparison of alirocumab or placebo in 18 924 patients who had an ACS 1 to 12 months previously and elevated atherogenic lipoproteins despite intensive statin therapy. Alirocumab dose was blindly titrated to target achieved low-density lipoprotein cholesterol (LDL-C) between 25 and 50 mg/dL. We examined the effects of treatment on all-cause death and its components, cardiovascular and noncardiovascular death, with log-rank testing. Joint semiparametric models tested associations between nonfatal cardiovascular events and cardiovascular or noncardiovascular death. Results: Median follow-up was 2.8 years. Death occurred in 334 (3.5%) and 392 (4.1%) patients, respectively, in the alirocumab and placebo groups (hazard ratio [HR], 0.85; 95% CI, 0.73 to 0.98; P=0.03, nominal P value). This resulted from nonsignificantly fewer cardiovascular (240 [2.5%] vs 271 [2.9%]; HR, 0.88; 95% CI, 0.74 to 1.05; P=0.15) and noncardiovascular (94 [1.0%] vs 121 [1.3%]; HR, 0.77; 95% CI, 0.59 to 1.01; P=0.06) deaths with alirocumab. In a prespecified analysis of 8242 patients eligible for ≥3 years follow-up, alirocumab reduced death (HR, 0.78; 95% CI, 0.65 to 0.94; P=0.01). Patients with nonfatal cardiovascular events were at increased risk for cardiovascular and noncardiovascular deaths (P<0.0001 for the associations). Alirocumab reduced total nonfatal cardiovascular events (P<0.001) and thereby may have attenuated the number of cardiovascular and noncardiovascular deaths. A post hoc analysis found that, compared to patients with lower LDL-C, patients with baseline LDL-C ≥100 mg/dL (2.59 mmol/L) had a greater absolute risk of death and a larger mortality benefit from alirocumab (HR, 0.71; 95% CI, 0.56 to 0.90; Pinteraction=0.007). In the alirocumab group, all-cause death declined wit h achieved LDL-C at 4 months of treatment, to a level of approximately 30 mg/dL (adjusted P=0.017 for linear trend). Conclusions: Alirocumab added to intensive statin therapy has the potential to reduce death after acute coronary syndrome, particularly if treatment is maintained for ≥3 years, if baseline LDL-C is ≥100 mg/dL, or if achieved LDL-C is low. Clinical Trial Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT01663402

ПРИНЦИПИ КОНСЕРВАТИВНОЇ ТЕРАПІЇ З УРАХУВАННЯМ ЛОКАЛІЗАЦІЇ ЕНДОМЕТРІОЇДНИХ ГЕТЕРОТОПІЙ

Introduction. In work the principles of conservative therapy of patients taking into account localization of endometriotic heterotopias, features of immune and hormonal homeostasis are represented.The study of the mechanisms genital endometriosis of pathogenic of different forms and degrees distribution was the purpose of work and development of rational medical tactic of the efficiency of medical treatment and rehabilitation of patients directed on the increase.Materials and methods. 115 patients with I-II outward and internal endometriosis are inspected, III-IV degrees of distribution. to 57 (49,6%) patients traditional conservative medical treatment of endometriosis was conducted. 58 (50,4%) patients got the differentiated hormonal and immunological medical treatment taking into account localization of endometriosis.Research results. The conducted researches showed that violations in a state of general immunity at patients with different clinical forms of endometriosis carried the second character, what the dynamics of growth of changes in immune status specifies on by a patient with growth of degree of distribution of disease, and also presence of substantial differences in general immunological reactivity depending on localization of endometriosis. Hormonal local homeostasis also has certain differences depending on localization and degree of weight of process, and it is needed to take into account during conducting of pathogenesis medical treatment of endometriosis with different localization.Conclusions. The conducted study of therapeutic efficiency of the differentiated medical treatment taking into account localization and degree of distribution of process showed more rapid decline of pain syndrome, and also more early renewal of reproductive function. Conservative therapy of endometriosis was conducted taking into account localization of endometriotic heterotopias of features of homeostasis (immune and hormonal) results in a clinical effect more early and stable, on comparison with traditional therapy, is instrumental in reduction of relapses of endometriosis in 1,5-2 times.В работе представлены принципы консервативной терапии больных с учетом локализации эндометриоидных гетеротопий, особенностей иммунного и гормонального гомеостаза.Целью работы явилось изучение механизмов патогенеза различных форм и степеней распространения генитального эндометриоза и разработка рациональной лечебной тактики, направленной на повышение эффективности лечения и реабилитации больных.Материалы и методы. Обследовано 115 больных наружным и внутренним эндометриозом I–II, III–IV степени распространения. 57 (49,6%) пациенткам проводилось традиционное консервативное лечение эндометриоза. 58 (50,4%) пациенток получали дифференцированное гормональное и иммунологическое лечение с учетом локализации эндометриоидных гетеротопий.Результаты исследования. Проведенные исследования показали, что нарушения в состоянии общего иммунитета у больных различными клиническими формами эндометриоза носят вторичный характер, на что указывает динамика нарастания изменений в иммунном статусе больных с ростом степени распространения заболевания, а также наличие существенных отличий в общей иммунологической реактивности в зависимости от локализации эндометриоидных гетеротопий. Гормональный местный гомеостаз также имеет определенные отличия в зависимости от локализации и степени тяжести процесса и это необходимо учитывать при проведении патогенетического лечения эндометриоза различной локализации.Выводы. Проведенное изучение терапевтической эффективности дифференцированного лечения с учетом локализации и степень распространения процесса показало более быстрое снижение болевого синдрома, а также более раннее восстановление репродуктивной функции. Консервативная терапия эндометриоза, проводимая с учетом локализации эндометриоидных гетеротопий, особенностей иммунного и гормонального гомеостаза, приводит к более раннему и стабильному клиническому эффекту, по сравнению с традиционной терапией, способствует уменьшению рецидивов эндометриоза в 1,5-2 раза.У роботі представлені принципи консервативної терапії хворих з урахуванням локалізації ендометріоїдних гетеротопій, особливостей імунного і гормонального гомеостазу.Метою роботи є вивчення механізмів патогенезу різних форм і ступенів розповсюдження генітального ендометріозу і розробка раціональної лікувальної тактики, направленої на підвищення ефективності лікування і реабілітації хворих.Матеріали і методи. Обстежено 115 хворих на зовнішній і внутрішній ендометріоз I-II, III-IV ступені розповсюдження. 57 (49,6%) пацієнткам проводилося традиційне консервативне лікування ендометріозу. 58 (50,4%) пацієнток одержували диференційоване гормональне і імунологічне лікування з урахуванням локалізації ендометріоїдних гетеротопій.Результати дослідження. Проведені дослідження показали, що порушення в стані загального імунітету у хворих на різні клінічні форми ендометріозу носять вторинний характер, на що вказує динаміка наростання змін в імунному статусі хворих із зростанням ступеня розповсюдження захворювання, а також наявність істотних відмінностей в загальній імунологічній реактивності залежно від локалізації ендометріоїдних гетеротопій. Гормональний місцевий гомеостаз також має певні відмінності залежно від локалізації і ступеня тяжкості процесу, і це необхідне враховувати при проведенні патогенетичного лікування ендометріозу різної локалізації.Висновки. Проведене вивчення терапевтичної ефективності диференційованого лікування з урахуванням локалізації і ступеня розповсюдження процесу показало більш швидке зниження больового синдрому, а також більш раннє відновлення репродуктивної функції. Консервативна терапія ендометріозу, яка проводилася з урахуванням локалізації ендометріоїдних гетеротопій, особливостей імунного і гормонального гомеостазу приводить до більш раннього і стабільного клінічного ефекту, в порівнянні з традиційною терапією, сприяє зменшенню рецидивів ендометріозу в 1,5-2 рази

IDENTIFICATION OF ANTIMICROBIAL COMPOUNDS FROM STREPTOMYCES SP. 156A BY CHROMATOGRAPHY AND MASSS

Bacterial strain 156A isolated from fresh water of Lake Baikal produces antibiotic compounds-homologies. The compounds have been isolated, purified and identified as nactins. The antibiotic mixture yield is 2 mg/l. The mixture consists of nonactin, monactin, dinactin, trinactin, tetractin and macrotetrolide C