Експериментальна модель технології комбінування антибактеріальних препаратів, що забезпечують антимікробну дію для лікування захворювань стафілококової етіології

У статті розглядаються створена експериментальна модельна система комбінованої дії антибіотиків природного походження (бензилпеніциліну) та напівсинтетичного (ампіоксу) з додаванням в якості допоміжної речовини кальція глюконату потенціюючи дію додаванням біополімера (ЕАР) Staphylococcus aureus. Спостерігали синергізм дії в комбінованих препаратах антибіотиків, глюконату кальція та біополімеру стафілокока. Показано, що застосування комбінації препаратів призводило до посилення антибактеріальної дії антибіотиків відносно Staphylococcus aureus, збільшення фагоцитарної активності та індексу цитотоксичності в модельній системі. Отримані результати можуть ввійти в основу розробки комбінованих препаратів з антибактеріальною дією та імунокорегуючими впливами, що можуть бути застосовані для лікування захворювань стафілококової етіології.The article discusses the design of an experimental model system which combines natural (Benzylpenicillin) and semi-synthetic (Ampiox) antibiotics with calcium gluconate as an auxiliary substance and their potentiation by addition of cell biopolymer of Staphylococcus aureus (EAP). The combination of antibiotics, calcium gluconate and staphylococcal biopolymer had a synergistic effect. It was shown that the drug combination results in enhancement of antimicrobial activity, as well as an increase of phagocytic activity and cytotoxicity index in the model system. The obtained results may form the design basis of the combined drugs which have antibacterial and immunocorrective effects that can be used in treatment staphylococcal caused diseases

Одержання моделей комбінованих імунобіологічних препаратів, що володіють противірусною та імунокорегуючою дією для забезпечення протективного імунітету

У статті розглянуто імунобіологічна активність людського гранулоцитарного колоніє стимулюючого фактора Г-КСФ (Філстим), інтерферону α-2b рекомбінантного (Лаферобіон), Герпевіру (міжнародна назва Ацикловір) та нанорозмірного біополімеру клітинної стінки Staphylococcus aureus. Також проаналізовано впливи моделей комбінованих імунобіологічних препаратів для забезпечення протективного імунітету та оптимізацію процесу виробництва препарату, що володіє противірусною, імуномодулюючою та імунокорегуючою дією. Показано, що при комбінованому застосуванні досліджуваного біополімера стафілокока (білку-адгезину ЕАР) та фармацевтичних препаратів спостерігали синергізм дії по імуномодулюючим властивостями, продукції захисних цитокінів та забезпеченні протективного імунітету. Отже отриманий нанорозмірний біополімер може бути використаний для моделювання біофармацевтичних препаратів з подібними властивостями.The article discusses the immunobiological activity of human granulocyte colony stimulating factor (G-CSF) (Filsm), recombinant interferon α-2b (Laferobion), Herpevir (international name Acyclovir) and nano-sized cell wall biopolymer of Staphylococcus aureus. We analyzed the effect of the combinations of these drugs on the development of protective immunity and conducted optimization of the drug production process to achieve antiviral, immunomodulatory and immuneregulating effects. Application of the staphylococcal adherence protein (EAP) biopolymer in combination with aforementioned drugs resulted in synergistic immunomodulatory activity, production of protective cytokines, and enhancement of protective immunity. Thus, a nano-sized biopolymer can be used to potentiate other drugs with similar properties

Somatic mutations of KIT in familial testicular germ cell tumours

Somatic mutations of the KIT gene have been reported in mast cell diseases and gastrointestinal stromal tumours. Recently, they have also been found in mediastinal and testicular germ cell tumours (TGCTs), particularly in cases with bilateral disease. We screened the KIT coding sequence (except exon 1) for germline mutations in 240 pedigrees with two or more cases of TGCT. No germline mutations were found. Exons 10, 11 and 17 of KIT were examined for somatic mutations in 123 TGCT from 93 multiple-case testicular cancer families. Five somatic mutations were identified; four were missense amino acid substitutions in exon 17 and one was a 12bp in-frame deletion in exon 11. Two of seven TGCT from cases with bilateral disease carried KIT mutations compared with 3 out 116 unilateral cases (p = 0.026). The results indicate that somatic KIT mutations are implicated in the development of a minority of familial as well as sporadic TGCT. They also lend support to the hypothesis that KIT mutations primarily take place during embryogenesis such that primordial germ cells with KIT mutations are distributed to both testes

Genome-wide linkage screen for testicular germ cell tumour susceptibility loci

A family history of disease is a strong risk factor for testicular germ cell tumour (TGCT). In order to identify the location of putative TGCT susceptibility gene(s) we conducted a linkage search in 237 pedigrees with two or more cases of TGCT. One hundred and seventy-nine pedigrees were evaluated genome-wide with an average inter-marker distance of 10 cM. An additional 58 pedigrees were used to more intensively investigate several genomic regions of interest. Genetic linkage analysis was performed with the ALLEGRO software using two model-based parametric analyses and a non-parametric analysis. Six genomic regions on chromosomes 2p23, 3p12, 3q26, 12p13-q21, 18q21-q23 and Xq27 showed heterogeneity LOD (HLOD) scores of greater than 1, with a maximum HLOD of 1.94 at 3q26. Genome-wide simulation studies indicate that the observed number of HLOD peaks greater than one does not differ significantly from that expected by chance. A TGCT locus at Xq27 has been previously reported. Of the 237 pedigrees examined in this study, 66 were previously unstudied at Xq27, no evidence for linkage to this region was observed in this new pedigree set. Overall, the results indicate that no single major locus can account for the majority of the familial aggregation of TGCT, and suggests that multiple susceptibility loci with weak effects contribute to the diseas

Прогностическое значение PD-L1-статуса опухоли у больных метастатическим раком предстательной железы

Background.New potential biomarker for patients with metastatic hormone-naive prostate cancer (PCa) might be detection of programmed death ligand 1 (PD-L1) expression in tumor which is associated with worsened results of treatment and decreased survival in patients with pancreatic cancer, lung cancer and other malignant tumors.Objective: to evaluate the prognostic value of positive tumor PD-L1 status on time to castration resistance (CRPCa) in patients with meta­static PCa receiving hormonal androgen deprivation therapy in first-line systemic treatment.Materials and methods.A total of 35patients with metastatic hormone-naive PCa receiving androgen deprivation therapy with luteinizing hormone-releasing hormone analogue and follow-up at N.N. Blokhin National Medical Research Center of Oncology were recruited in our prospective study. Tumor features of all patients were evaluated for PD-L1 expression on tumor cells by immunohistochemical studies of paraffin block sections obtained under the visual control of the pathologist using a set of monoclonal anti-PD-L1 antibody (28-8) (ab 205921) and Ventana BenchMark GXSlide staining system. Tumor tissue was obtained before starting androgen deprivation therapy. The expression level of PD-L1 >1 % in tumor cells was taken for the positive tumor PD-L1(+) status.Results. Median follow-up was 32.8 months. Positive tumor PD-L1(+) status was identified in 10 (28.6 %) cases. Median time to CRPCa was significantly lower in patients with PD-L1(+) status, than in negative PD-L1(—) status (21.44 vs. 49.12, p = 0.006 log rank test). Multi­variate Cox regression analysis confirmed independence prognostic value of PD-L1(+) associated with decreased time to CRPCa (hazard ration 5.95, 95 % confidence interval 1.97—17.99; p = 0.002), including in subgroup of patients with low-volume metastatic disease (hazard ration 7.33, 95 % confidence interval 1.81—29.60; p = 0.005).Discussion. Interaction of PD-1 receptors and its ligands PD-L1/PD-L2 is the key mechanism causing tumor immune escape and progression of the cancer. There are discussed certain ways of inducing PD-L1 expression and its prognostic value on aggressive nonmetastatic PCa. High frequency of positive PD-L1 status was revealed in rare histological subtypes of PCa associated with unfavorable prognosis and visceral metastasis.Conclusion.The results of our study demonstrated the positive tumor PD-L1 status as an independent unfavorable prognostic factorfor patients with metastatic hormone-naive PCa associated with decreased time to castration resistance, including in patients with low volume metastatic disease.Введение. Одним из потенциальных биомаркеров для больных метастатическим гормоночувствительным раком предстательной железы (РПЖ) может быть определение экспрессии лиганда белка программируемой клеточной гибели (PD-L1) в опухоли, ассо­циированной с неблагоприятными результатами лечения и снижением выживаемости больных раком поджелудочной железы, легкого и другими злокачественными новообразованиями.Цель исследования — оценка прогностической значимости положительного статуса PD-L1(+) опухоли на время до развития кастрационной резистентности (КРРПЖ) у больных метастатическим РПЖ, получающих гормональную андрогендепривационную терапию в 1-й линии системного противоопухолевого лечения.Материалы и методы. В проспективный анализ были включены данные 35пациентов с метастатическим гормоночувствительным РПЖ, которым проводилась андрогендепривационная терапия аналогами лютеинизирующего гормона рилизинг-гормона и которые находились под наблюдением в НМИЦонкологии им. Н.Н. Блохина. Всем пациентам было проведено определение экспрессии PD-L1 в опухолевых клетках с применением метода иммуногистохимического исследования срезов парафиновых блоков, полученных под контролем патоморфолога с использованием моноклонального антитела Anti-PD-L1 antibody (28-8) (ab 205921) на иммуностейнере Ventana BenchMark GX. Опухолевый материал был получен до начала андрогендепривационной терапии у пациентов. За статус PD-L1(+) принимали уровень экспрессии PD-L1 >1 % в опухолевых клетках.Результаты. Медиана наблюдения составила 32,8 мес. Статус PD-L1(+) опухоли подтвержден в 10 (28,6 %) случаях. Медиана времени до КРРПЖ была достоверно ниже в группе PD-L1(+), чем в группе PD-L1(—) (21,44 мес против 49,12 мес; р = 0,006). Многофакторный анализ Кокса подтвердил PD-L1(+) как независимый фактор прогноза, ассоциированный со снижением времени до КРРПЖ (отношение рисков 5,95, 95 % доверительный интервал 1,97—17,99; р = 0,002), в том числе в подгруппе больных с незначительной распространенностью метастатического поражения (отношение рисков 7,33, 95 % доверительный интервал 1,81—29,60; р = 0,005).Обсуждение. Взаимодействие рецептора PD-1 с его лигандами PD-L1/PD-L2 является ключевым механизмом в «ускользании» опухоли от иммунологического противоопухолевого надзора. Приведены различные механизмы активации экспрессии PD-L1, а также ее связь с агрессивным фенотипом при неметастатическом РПЖ. Высокая частота положительного статуса PD-L1 обнаружена при редких неблагоприятных гистологических формах РПЖ и висцеральных метастазах.Заключение. Результаты исследования показали, что положительный статус PD-L1 опухоли является независимым фактором неблагоприятного прогноза для больных метастатическим гормоночувствительным РПЖ, ассоциированным со снижением времени до развития КРРПЖ, в том числе при минимальной распространенности метастатического поражения

Modern fluorescent techniques to investigate the mechanisms of lymphocyte activation

Fluorescent proteins are promising tools for studying intracellular signaling processes in lymphocytes. This brief review summarizes fluorescence-based imaging techniques developed in recent years and discusses new methodological advances, such as fluorescent photoswitches, fluorescence recovery after photobleaching (FRAP), fluorescent resonance energy transfer (FRET), fluorescence lifetime imaging microscopy (FLIM), photoactivated localization microscopy (PALM), stochastic optical reconstruction microscopy (STORM), stimulated emission depletion (STED), total internal reflection fluorescence (TIRF) and other techiques. This survey also highlights recent advances in vitro imaging of live tissues, novel applications of flow cytometry with genetically modified fluorescent proteins, and future prospects for the development of new immunological test systems based on fluorescent protein technology

The prognostic value of tumor PD-L1 status in patients with metastatic prostate cancer

Background.New potential biomarker for patients with metastatic hormone-naive prostate cancer (PCa) might be detection of programmed death ligand 1 (PD-L1) expression in tumor which is associated with worsened results of treatment and decreased survival in patients with pancreatic cancer, lung cancer and other malignant tumors.Objective: to evaluate the prognostic value of positive tumor PD-L1 status on time to castration resistance (CRPCa) in patients with meta­static PCa receiving hormonal androgen deprivation therapy in first-line systemic treatment.Materials and methods.A total of 35patients with metastatic hormone-naive PCa receiving androgen deprivation therapy with luteinizing hormone-releasing hormone analogue and follow-up at N.N. Blokhin National Medical Research Center of Oncology were recruited in our prospective study. Tumor features of all patients were evaluated for PD-L1 expression on tumor cells by immunohistochemical studies of paraffin block sections obtained under the visual control of the pathologist using a set of monoclonal anti-PD-L1 antibody (28-8) (ab 205921) and Ventana BenchMark GXSlide staining system. Tumor tissue was obtained before starting androgen deprivation therapy. The expression level of PD-L1 >1 % in tumor cells was taken for the positive tumor PD-L1(+) status.Results. Median follow-up was 32.8 months. Positive tumor PD-L1(+) status was identified in 10 (28.6 %) cases. Median time to CRPCa was significantly lower in patients with PD-L1(+) status, than in negative PD-L1(—) status (21.44 vs. 49.12, p = 0.006 log rank test). Multi­variate Cox regression analysis confirmed independence prognostic value of PD-L1(+) associated with decreased time to CRPCa (hazard ration 5.95, 95 % confidence interval 1.97—17.99; p = 0.002), including in subgroup of patients with low-volume metastatic disease (hazard ration 7.33, 95 % confidence interval 1.81—29.60; p = 0.005).Discussion. Interaction of PD-1 receptors and its ligands PD-L1/PD-L2 is the key mechanism causing tumor immune escape and progression of the cancer. There are discussed certain ways of inducing PD-L1 expression and its prognostic value on aggressive nonmetastatic PCa. High frequency of positive PD-L1 status was revealed in rare histological subtypes of PCa associated with unfavorable prognosis and visceral metastasis.Conclusion.The results of our study demonstrated the positive tumor PD-L1 status as an independent unfavorable prognostic factorfor patients with metastatic hormone-naive PCa associated with decreased time to castration resistance, including in patients with low volume metastatic disease