THE SENSITIVITY OF DEVELOPED ELISA TEST SYSTEM FOR AFLATOXIN B1 DETECTION COMPARED TO CHROMATOGRAPHIC METHODS

Розроблено тест-систему для виявлення афлатоксину В1 методом ІФА. Відпрацьована методика синтезу імуноферментного кон’югату афлотоксин-ЛФ і підібрано стабілізуючі розчини для його зберігання та створення імуноферментної тест-системи. Підібрано стабілізуючий розчин для зберігання кон’югату та розчин на основі трісового буфера, який найкраще справлявся з подоланням неспецифічних взаємодій антитіл з матеріалом планшету і зменшував кількість хибних результатів при постановці аналізу.Для встановлення чутливості розробленої ІФА тест-системи готували серію розведень стандарту афлатоксину В1 в молоці з концентраціями 0, 1, 5, 10, 20, 40,  80, 160, 320, 640 ppb. Зразки тестували методом ІФА та за допомогою ВЕРХ. Визначено, що чутливість виявлення афлатоксину В1 в розробленій тест-системі ІФА (ELISA) складає 4-40 ppb, а у ВЕРХ – 1-320 ppb. Доведено, що хроматографічні методи (тонкошарова хроматографія та ВЕРХ) менш специфічні порівняно з імуноферментним аналізом. Отримавши основні компоненти тест-системи і оптимізувавши умови проведення аналізу показали, що розроблена тест-система легко відтворювана, має високу чутливість та специфічність.Разработана тест-система для опредеения афлатоксина В1 методом ИФА. Отработанная методика синтеза иммуноферментного конъюгата афлотоксин-ЩФ и подобраны стабилизирующие растворы для его хранения для создания иммуноферментной тест-системы. Подобран стабилизирующий раствор для хранения конъюгата и раствор на основе трисового буфера, который лучше справлялся с преодолением неспецифических взаимодействий антител с материалом планшета и уменьшал количество ложных результатов при постановке анализа.Для определения чувствительности разработанной ИФА тест-системы готовили серию разведений стандарта афлатоксина В1 в молоке с концентрациями 0, 1, 5, 10, 20, 40, 80, 160, 320, 640 ppb. Образцы тестировали методом ИФА и с помощью ВЭЖХ. Определено, что чувствительность обнаружения афлатоксина В1 в разработаной тест-системе ИФА (ELISA) составляет 4-40 ppb, а в ВЭЖХ – 1-320 ppb. Доказано, что хроматографические методы (тонкослойной хроматографии и ВЭЖХ) менее специфичны по сравнению с иммуноферментным анализом. Получив основные компоненты тест-системы и оптимизировав условия проведения анализа показано, что разработанная тест-система легко воспроизводима, имеет высокую чувствительность и специфичность.The test system was developed for the detection of aflatoxin B1 by ELISA. The methodology for synthesis of immune-enzyme aflatoxin-alkaline phosphatase conjugates was completed and selected stabilizing solutions for the storage and creating ELISA test system. The stabilizing solution was selected for storing conjugate and trises buffer solution that better cope with overcoming nonspecific interactions of antibodies with the material of plate and reduces amount of false positives in the formulation of analysis. For determining the sensitivity of developed ELISA test systems was prepared the dilutions series of aflatoxin B1 standards in milk at the concentrations 0, 1, 5, 10, 20, 40, 80, 160, 320, 640 ppb. The samples were analized by ELISA and HPLC. Determined that the sensitivity of detection of aflatoxin B1 at the developed of ELISA test system is 4-40 ppb, and HPLC - 1-320 ppb. Was proved that the chromatographic techniques (thin layer chromatography and HPLC) compared with the ELISA less specific. After receiving the basic components of the test system and optimized of the analysis conditions were showed that the developed test system can be easily reproducible and has a high sensitivity and specificity

Clinical Experience of Luminescent Diagnostics of Precancerous Diseases and Cervical Cancer

The article summarizes the experience of using luminescent diagnostics with the use of ytterbium porphyrin complexes in gynecology and oncology. A pharmaceutical composition based on the Yb complex of 2,4-dimethoxyhematoporphyrin IX was used as the luminescent markers within the infrared range. The determination of luminescence characteristics (luminescence intensity) was carried out using a laserfiber fluorimeter in the range of 900-1100 nm. A new method for diagnosis of cervical disease has been proposed. The method of luminescent diagnostics allows to conduct a survey of a large number of patients in a short time. The method of luminescent diagnostics using the ytterbium complexes of porphyrins is not invasive. The method can be used as a screening. Differences between normal and pathologically altered cervical tissue have been identified and differences between pathological changes in the cervix HSIL (CIN II, CIN III) and cervical cancer are reliable. Keywords: Cervical cancer, squamous cell carcinoma, diagnosis of cervical cancer, squamous intraepithelial lesions of high grade – HSIL, luminescent diagnostics, luminescing in the near infrared (NIR) spectral region, porphyrins, ytterbium complexes of porphyrins

МОРФОМЕТРИЧЕСКОЕ ИССЛЕДОВАНИЕ ПЕЧЕНИ КРЫС ВИСТАР С МОДЕЛЬЮ АЛИМЕНТАРНОГО ОЖИРЕНИЯ

Experimental alimentary obesity leads to significant structural changes in the liver of rats. Structural changes in the parenchymal cells are accompanied by functional strain in the capillary-connective tissue structures, and by disorders in the blood circulation and lymph drainage in the liver.Экспериментальное алиментарное ожирение приводит к значительным структурным изменениям в печени крыс. Структурные изменения в паренхиматозных клетках сопровождаются функциональным напряжением капилляросоединительнотканных структур, нарушением кровообращения и лимфотока в печени

LOW PROFILE SLOT ANTENNA INTEGRATED WITH SOLAR PANEL

The integration of solar panels with antennas allows you to achieve communication in remote areas to provide the capabilities of the Internet of things, monitoring of vital signs of the body. The paper deals with the integration of a low-profile corrugated antenna into a solar panel design

НОВЫЕ ТЕХНОЛОГИИ ВОССТАНОВЛЕНИЯ И ЗАЩИТЫ ЭНЕРГЕТИЧЕСКОГО ОБОРУДОВАНИЯ КОМПОЗИТНЫМИ МАТЕРИАЛАМИ

The analysis of possible variants of reconstruction of the power equipment is fulfilled and the conclusion concerning the prospects of such work with the use of composite materials is reached. The data on the technical characteristics of composite repair materials for various purposes are presented, the results of repairs of power equipment, in particular the technology for the recovery of the boarding surfaces of the diffuser rings and protection of the pumps D1250 casings are provided. The technology of the recovery pneumatic cylinder, hydraulic cylinder rod, as well as the unique technology of restoration of working surfaces of the impeller vanes of transfer pump, that had been destroyed by corrosion in conjunction with the cavitation processes and were considered as not restorable is described. The restored impeller was in operation during a year and only thereafter it was removed for restoration. Another composite material discussed in the article – diagum – makes it possible to perform a series of repairs associated with restoration of the rubber-covered surfaces of pump casings as well as with restoration of various surfaces of the conveyor belts. Taking the excellent adhesive properties of this composite into account, restoration of worn stainless steel sieve screens to remove abrasive material was fulfilled with the aid of it. The restoration was accomplished via the use of the conveyor belt which application time had expired, that was glued to a metal sieve with diagum. The use of the composites is economically justified, because the application of them in repairs reduces, firstly, terms of restoration work and, secondly, the price of repairs. Third, equipment that was damaged beyond repair is being commissioned by the use of the mentioned composites. Проведен анализ возможных вариантов восстановления энергетического оборудования и сделан вывод о перспективности выполнения таких работ с помощью композитных материалов. Изложены сведения о технических характеристиках композитных ремонтных материалов различного назначения, приведены результаты ремонтов энергетического оборудования, в частности отработана технология восстановления посадочных поверхностей под диффузорными кольцами и защиты корпусов насосов Д1250. Описана технология восстановления пневмоцилиндра, штока гидроцилиндра, а также уникальная технология восстановления рабочих поверхностей лопаток колеса перекачивающего насоса, которые были полностью сведены коррозией в совокупности с кавитационными процессами и восстановлению традиционными способами не подлежали. Восстановленное колесо отработало один год, и лишь после этого было снято на повторное восстановление. Еще один композитный материал из рассмотренных в статье – диагум – позволил выполнить целую серию ремонтных работ, связанных как с восстановлением обрезиненных поверхностей корпусов насосов, так и с ремонтом различного вида поверхностей транспортерных лент. Учитывая отличные адгезионные свойства данного композита, с его помощью выполнено восстановление изношенных нержавеющих сит грохотов для отсева абразивного материала. Это восстановление выполняли за счет использования отслужившей свой срок транспортерной ленты, которую приклеивали к металлическому ситу с помощью диагума. Применение композитов экономически оправдано, поскольку благодаря им сроки восстановительных работ сокращаются, снижается себестоимость ремонтов и вводится в строй оборудование, которое восстановлению не подлежало

Common variable immunodeficiency disorder: a clinical case

Primary immunodeficiency is a rare congenital pathology associated with failure of immune system, manifested by disturbances of its functions. These defects lead to increased susceptibility of patients to various infectious agents, as well as the development of autoimmune, malignant and other diseases. Primary immunodeficiency is classified as a rare disease, which was previously associated with a poor prognosis with a high risk of mortality in childhood. To date, the emergence of highly effective treatment methods has changed the course and prognosis of these diseases. Clinicians of various specialties increasingly meet with this pathology in everyday practice, including adult age cohorts. In this regard, early diagnosis of primary immunodeficiency in adults becomes relevant, being associated with choosing optimal therapy, prevention of severe internal organ damage, determination of management strategy for the patient, as well as the need to identify inherited disorders and provide information to the patient’s family. Delayed verification of the diagnosis may cause disability of the patient and development of irreversible, often fatal complications. This article presents our own clinical case with a newly diagnosed clinical condition: Common variable immunodeficiency disorder (CVID), the most common form of primary immunodeficiency in adults. The symptoms of common variable immunodeficiency disorder appear in these patients in adulthood, but a high-quality collected history of the disease will allow you to trace symptoms in the patients even since early childhood. There is a common gap for several years between the onset of the disease and clinical diagnosis, since erroneous diagnosis is often made due to non-specific clinical symptoms that resemble other, more frequent diseases. The prognosis of patients with CVID depends on several factors: frequency of infections, structural disorders in the lungs, the occurrence of autoimmune diseases and the success of infection prevention. Thus, a variety of clinical forms of primary immunodeficiency, lack of awareness of doctors about this pathology, complexity of immunological examination in the general medical network lead to the fact that CVID is not diagnosed for long terms, and patients do not receive the necessary pathogenetic therapy. There is a need for drawing attention of doctors of various disciplines to the fact that the recurrent inflammatory processes of various localization, which are difficult to respond to adequate traditional therapy, may be caused by changes in the immune system, including congenital, genetically determined immunodeficiency

Сучасний стан захворюваності на злоякісні новоутворення порожнини рота в Полтавській області за 2004-2010 роки

В роботі представлений стан онкологічної захворюваності порожнини рота в Полтавському регіоні за 7 років, з 2004- 2010 роки. Виявлена висока занедбаність пухлин цієї локалізації. За статистикою просліджується висока смертність до 1 року з моменту встановлення діагнозу. Вона становить 29,8%. Виявлена велика кількість відмов від лікування на всіх етапах. Зроблені висновки стосовно причин пізньої пізньої діагностики цієї локалізації. Загальна кількість хворих на злоякісні новоутворення порожнини рота в Полтавській області за 2004-2010 роки становила 794 особи, із них в місті проживають 214 чоловік (26,95%), у сільській місцевості – 580 (73,04%) хворих, (що у 2,71 рази більше, ніж у місті), за статтю хворих розподілили : жінок, вражених онкопатологією, було – 174 (22,16%), чоловіків спостерігалося – 618 (77,84%), що у 3,55 рази перевищує кількість жінок, смертність до року становила – 306 випадків. За віком хворих розподілили таким чином: 1група (до 45 років) – 8 осіб, 2 група (45-59 років) – 160 випадків, найбільша кількість хворих зафіксована у 3-й віковій групі (60-74 роки) - 350 людей, 4 група (75-89 років) - 215 осіб, 5 група (>90 років) - зафіксовано 25 випадків захворювання. На першому місці за кількістю хворих стоїть рак губ – 355 людей, друге місце належить ЗН дна порожнини рота (147 випадків), інших та не уточнених частин язика – 96 хворих, пухлини язика - 75 людей, піднебіння – 67 осіб, останнє місце займають ЗН ясен - 54 особи. Ключові; В работе представлен состояние онкологической заболеваемости полости рта в Полтавском регионе за 7 лет, с 2004-2010 года. Обнаружена высокая запущенность опухолей этой локализации. По статистике прослеживается высокая смертность до 1 года с момента установления диагноза. Она составляет 29,8%. Обнаружено большое количество отказов от лечения на всех этапах. Сделанные выводы относительно причин поздней поздней диагностики этой локализации. Общее количество больных злокачественными новообразованиями полости рта в Полтавской области за 2004-2010 годы составила 794 человека, из них в городе проживают 214 человек (26,95%), в сельской местности - 580 (73,04%) больных, (что в 2,71 раза больше, чем в городе), по полу больных распределили: женщин, пораженных онкопатологией, было - 174 (22,16%), мужчин наблюдалось - 618 (77,84%), что в 3,55 раза превышает количество женщин, смертность до года составляла - 306 случаев. По возрасту больных распределили следующим образом: 1 группы (до 45 лет) - 8 человек, 2 группа (45-59 лет) - 160 случаев, наибольшее число заболевших зафиксировано в 3-й возрастной группе (60-74 года) - 350 человек, 4 группа (75-89 лет) - 215 человек, 5 группа (> 90 лет) - зафиксировано 25 случаев заболевания. На первом месте по количеству больных стоит рак губ - 355 человек, второе место принадлежит ЗН дна полости рта (147 случаев), других но не уточненных частей языка - 96 больных, опухоли языка - 75 человек, небо - 67 человек, последнее место занимают ЗН десен - 54 человека. Ключевые; The paper presents the state of oncological morbidity of the oral cavity in the Poltava region for 7 years, from 2004-2010. The high neglect of tumors of this localization was revealed. According to statistics, a high mortality rate of up to 1 year from the time of diagnosis is detected. It is 29.8%. A large number of abandoned treatments have been detected at all stages. Conclusions are made regarding the reasons for the late late diagnosis of this localization. The total number of patients with malignant neoplasms of the oral cavity in the Poltava region in 2004-2010 was 794 people, of which 214 people live in the city (26.95%), in rural areas - 580 (73.04%) patients (which 2.71 times more than in the city), according to the gender of the patients: women who were affected by oncopathology were - 174 (22,16%), men were observed - 618 (77,84%), which is 3,55 times the number women, the death rate to the year was - 306 cases. By age, the patients were divided as follows: 1 group (up to 45 years old) - 8 people, 2 groups (45-59 years) - 160 cases, the largest number of patients recorded in the 3rd age group (60-74 years) - 350 people, 4 group (75-89 years old) - 215 people, 5 groups (> 90 years) - 25 cases of the disease were recorded. In the first place in the number of patients there is a cancer of the lips - 355 people, the second place belongs to the bottom of the oral cavity (147 cases), the other and unspecified parts of the tongue - 96 patients, tongue tongue - 75 people, palate - 67 people, the last place occupy ZN ash - 54 people. Keys

RNA-Seq gene expression profiling of HepG2 cells: The influence of experimental factors and comparison with liver tissue

© Tyakht et al.; licensee BioMed Central. Background: Human hepatoma HepG2 cells are used as an in vitro model of the human liver. High-throughput transcriptomic sequencing is an advanced approach for assessing the functional state of a tissue or cell type. However, the influence of experimental factors, such as the sample preparation method and inter-laboratory variation, on the transcriptomic profile has not been evaluated. Results: The whole-transcriptome sequencing of HepG2 cells was performed using the SOLiD platform and validated using droplet digital PCR. The gene expression profile was compared to the results obtained with the same sequencing method in another laboratory and using another sample preparation method. We also compared the transcriptomic profile HepG2 cells with that of liver tissue. Comparison of the gene expression profiles between the HepG2 cell line and liver tissue revealed the highest variation, followed by HepG2 cells submitted to two different sample preparation protocols. The lowest variation was observed between HepG2 cells prepared by two different laboratories using the same protocol. The enrichment analysis of the genes that were differentially expressed between HepG2 cells and liver tissue mainly revealed the cancer-associated gene signature of HepG2 cells and the activation of the response to chemical stimuli in the liver tissue. The HepG2 transcriptome obtained with the SOLiD platform was highly correlated with the published transcriptome obtained with the Illumina and Helicos platforms, with moderate correspondence to microarrays. Conclusions: In the present study, we assessed the influence of experimental factors on the HepG2 transcriptome and identified differences in gene expression between the HepG2 cell line and liver cells. These findings will facilitate robust experimental design in the fields of pharmacology and toxicology. Our results were supported by a comparative analysis with previous HepG2 gene expression studies

Genome-Wide Mycobacterium tuberculosis Variation (GMTV) Database: A New Tool for Integrating Sequence Variations and Epidemiology

Background Tuberculosis (TB) poses a worldwide threat due to advancing multidrug-resistant strains and deadly co-infections with Human immunodeficiency virus. Today large amounts of Mycobacterium tuberculosis whole genome sequencing data are being assessed broadly and yet there exists no comprehensive online resource that connects M. tuberculosis genome variants with geographic origin, with drug resistance or with clinical outcome. Description Here we describe a broadly inclusive unifying Genome-wide Mycobacterium tuberculosis Variation (GMTV) database, (http://mtb.dobzhanskycenter.org) that catalogues genome variations of M. tuberculosis strains collected across Russia. GMTV contains a broad spectrum of data derived from different sources and related to M. tuberculosis molecular biology, epidemiology, TB clinical outcome, year and place of isolation, drug resistance profiles and displays the variants across the genome using a dedicated genome browser. GMTV database, which includes 1084 genomes and over 69,000 SNP or Indel variants, can be queried about M. tuberculosis genome variation and putative associations with drug resistance, geographical origin, and clinical stages and outcomes. Conclusions Implementation of GMTV tracks the pattern of changes of M. tuberculosis strains in different geographical areas, facilitates disease gene discoveries associated with drug resistance or different clinical sequelae, and automates comparative genomic analyses among M. tuberculosis strains

Tumor inflating lymphocytes. Purification, expanding and cytotoxicity analisys on primary tumor cultures

Background. Tumor Infiltrating Lymphocytes (TILs) is one of the most promising sources of autologous cytotoxic T-cells for adoptive immunotherapy, which has already shown high efficiency in the treatment of metastatic melanoma. However, the isolation of TILs from solid tumors is technically difficult. A suppressive tumor microenvironment, in particular, a high level of expression of check-point inhibitors PD-1 CTLA4, tissue hypoxia and other factors cause that T cells isolated from the tumor do not proliferate well and do not exhibit cytotoxic properties. Aims. In this study, we isolated TILs from surgical material obtained by resection of solid tumors (primary and metastatic adenocarcinomas of various localization, melanoma, glioblastoma), studied their population composition and developed protocols for the purification expanding, and activation of CD4+, CD8+ cytotoxic antitumor lymphocytes. Methods. An urgent task is the activation of TILs, turning off immunosuppressive mechanisms and increasing their antitumor cytotoxic activity. Various approaches are used for this: activation by a cocktail of cytokines and antibodies, editing the lymphocyte genome by knocking out suppressor genes or, conversely, transduction of activating genes, coincubation with feeder cells, etc. Cells were obtained from samples of resected tumors in 16 patients; in each case we obtain an autologous pair: the primary tumor culture and the TILs culture. Results. We could isolate viable lymphocytes in 100% of cases. Isolated TILs were successfully expanded in our specialized medium using various combinations of IL-2, IL-15, IL-21, IL-7, anti-CD3 and anti-CD28. Immunophenotyping showed that the obtained TILs are a heterogeneous mixture of CD4+, CD8+ cells containing populations of CD3+CD8+CD45+(CTL) CD3+CD4+CD45+ (T-helpers), CD4+CD25+CD127- (Т-regulatory cells), CD3-CD56+CD45+ (NK-cells), CD3+CD56+CD45+ (Т-NK-cells). The initial cultures of TILs were also characterized by a high level of PD1 expression, indicating their low antitumor cytotoxicity. Using different protocols of isolation, expansion, and activation, we obtained a cell preparation containing 80% of CD8+ PD-1- activated TILs in an amount sufficient for adoptive therapy (500106 or more). An in vitro study of the cytotoxicity of obtained TILs in primary cultures of homologous tumors using RTCA Icelligence showed high cytotoxicity, providing almost 100% tumor cell death. Conclusion. Our developed protocol for the production and activation of TILs can be recommended for the phase III clinical trials of adoptive immunotherapy of recurrent, highly metastatic solid tumors