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Background: Schistosomiasis is one of the neglected tropical diseases with recent evidences about the high prevalence among preschool-age children. The pediatric formulation of Praziquantel (PZQ) has to be assessed for the efficacy as it gave controversial results in several countries. Objective(s): The current study aimed at evaluating the efficacy of the pediatric suspension of PZQ against Schistosoma mansoni Egyptian strain in the experimental animals. Methods: 150 Swiss albino mice infected with Schistosoma mansoni were divided into three groups, the first group was treated with 600 mg/kg body weight of PZQ pediatric suspension, the second group was treated with 600 mg/kg PZQ tablets and the third one received no treatment as a control. The efficacy of the pediatric formulation was experimentally evaluated in comparison with the tablet formulation as a benchmark on the basis of the following specific parasitological parameters (worm burden, tissue egg load, and oogram pattern i.e. percentage of dead, live or immature eggs shown in the stool sample). Results: The comparison between the mean egg count per gram stool in the two groups pediatric suspension of PZQ (Epiquantel) and adult tablets of PZQ (Distocide), and the control group by applying one way ANOVA revealed a statistically significant difference (p

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Antibodies forming a complex with antigen in vivo can dramatically change the antibody response to this antigen. In some situations, the response will be a 100-fold stronger than in animals immunized with antigen alone, and in other situations, the response will be completely suppressed. IgG is known to suppress the antibody response, for example to erythrocytes, and this is used clinically in Rhesus prophylaxis. The mechanism behind IgG-mediated immune suppression is still not understood. Here, we will review studies performed in experimental animal models and discuss the various hypotheses put forward to explain the profound suppressive effect of IgG. We conclude that an exclusive role for negative regulation of B cells through Fc gamma RIIB, increased clearance of erythrocytes from the circulation or complement-mediated lysis is unlikely. Epitope masking, where IgG hides the epitope from B cells, or trogocytosis, where IgG removes the epitope from the erythrocyte, is compatible with many observations. These two mechanisms are not mutually exclusive. Moreover, it cannot be ruled out that clearance, in combination with other mechanisms, plays a role.

The purpose of the study: to analize the dynamics of the concentration of IgG in thedevelopment of experimental ulcerous - necrotic gingivitis with and without the localtreatment by a complex of antioxidant preparations.Material and methods: the study was conducted on 18 rabbits. The model of ulcerous -necrotic gingivitis was obtained in animals by chemical burns. Experimentalpreparations were applied to the damaged gum area 2 times a day in 2 hours after feedingthe animals at approximate dose of 200 mg. The nature of the course of experimentalulcerous - necrotic gingivitis was investigated on the 3rd, 5th, 7th and 10th days of thehealing process.Results: On the 3rd day in control animals, IgG levels increased 50.0% above the level ofintact animals. On the 5th day, the prevalence over the index of physiological norm wasalready 30.0%. The indicated trend continued, although on the 7th day the data obtainedsignificantly differed from those of intact animals and prevailed 10.56%. Only at the endof the study (10th day), a result was obtained that statistically insignificantly differedfrom the indicator of intact animals and made it 104.45%.Conclusions: Local application of the developed complex of antioxidant preparationsallows to adjust the course of the inflammatory reaction in the damaged mucousmembrane of the oral cavity. As a result, moderate, less than in the control group, growthof the indexes of the markers studied in the initial phase and their complete normalizationhave been already seen on the 7th day of observation., Цель работы - проанализировать динамику концентрации IgG при развитии экспериментального язвенно - некротического гингивита без и на фоне местного лечениякомплексом антиоксидантных препаратов.Материалы и методы. Исследования проводились на 18-ти кроликах. Модельязвенно-некротического гингивита получали у животных путем химического ожога. Комплекс препаратов в ориентировочной дозе 200 мг наносили на поврежденный участок десны 2 раза в сутки через 2 часа после кормления животных.Исследовался характер течения экспериментального язвенно - некротическогогингивита на 3-и, 5-е, 7-е и 10-е сутки процесса заживления.Результаты. На 3-и сутки у животных контрольной группы наблюдался рост содержания IgG на 50,0% выше уровня интактных животных. На 5-е сутки преимущество над показателем физиологической нормы составляло уже 30,0%.Указанная тенденция сохранялась и в дальнейшем, хотя еще полученные на 7-е сутки данные достоверно отличались от данных интактных животных, и превосходили их на 10,56%. Только к окончанию исследования (10-е сутки) был полученрезультат, который статистически недостоверно отличался от показателяинтактных животных, и составлял 104,45%.Выводы. Местное применение разработанного комплекса антиоксидантных препаратов позволяет скорректировать ход воспалительной реакции на поврежденой слизистой оболочке полости рта. В результате отмечено умеренный, меньшечем в контрольной группе, рост показателей исследуемых маркеров в начальнойфазе, и их полноценная нормализация уже на 7-е сутки наблюдения., Мета роботи - проаназізувати динаміку концентрації IgG при розвитку експериментального виразково-некротичного гінгівіту без та на тлі місцевого лікуваннякомплексом антиоксидантних препаратів.Матеріали і методи. дослідження проводили на 18 кроликах. Модель виразковонекротичного гінгівіту отримували у тварин шляхом хімічного опіку. Дослідніпрепарати в орієнтовній дозі 200 мг наносили на пошкоджену ділянку ясен 2 разина добу через 2 год після годування тварин. Досліджували характер перебігу експериментального виразково-некротичного гінгівіту на 3-тю, 5-ту, 7-му та 10-тудоби процесу загоєння.Результати. На 3-тю добу у тварин контрольної групи спостерігали зростаннявмісту IgG на 50,0% над рівнем інтактних тварин. На 5-ту добу перевага над показником фізіологічної норми становила вже 30,0%. Вказана тенденція зберігаласяі надалі, хоч ще на 7-му добу отримані дані достовірно різнилися від данихінтактних тварин та переважали їх на 10,56%. Лише на закінчення дослідження(10-та доба) отримано результат, який статистично недостовірно відрізнявся відпоказника інтактних тварин та становив його 104,45%.Висновки. Місцеве застосуванням розробленого комплексу антиоксидантних препаратів дає змогу скорегувати перебіг запальної реакції на пошкодження слизовоїоболонки порожнини рота. Як наслідок, відзначено помірковане, менше ніж уконтрольній групі, зростання показників досліджуваних маркерів у початковій фазіта їх повноцінну нормалізацію вже на 7-му добу спостереження.

Patofiziološki procesi celjenja ran pri hiperglikemiji so zelo kompleksni in še niso dovolj pojasnjeni. Dipeptidil-peptidaza IV ali molekula CD26 (DPP IV/CD26) je multifunkcionalen protein, ki je v telesu prisoten v številnih celicah in različnih bioloških tekočinah. Poleg tega, da je vključen v vzdrževanje homeostaze glukoze z uravnavanjem biološke aktivnosti različnih substratov, igra pomembno vlogo tudi v procesih angiogeneze ter proliferacije, migracije in apoptoze različnih vrst celic, pomembnih za celjenje poškodovanega oz. ranjenega tkiva. Podatki iz literaure kažejo na to, da inhibicija DPP IV/CD26 prispeva k boljšemu celjenju ran pri bolnikih s sladkorno boleznijo. Namen magistrske naloge je bil vzpostaviti eksperimentalni model diabetesa (eksperimentalne hiperglikemije) v divjem tipu (C57BL/6) in CD26 deficientnih (CD26-/-) miših, ter ugotoviti ali, in na kakšen način, v takih okoliščinah pomanjkanje DPP IV/CD26 vpliva na procesa angiogeneze in regeneracije kožnega tkiva. Razvoj diabetesa v poskusnih živalih smo spremljali s pomočjo kliničnih in biokemičnih parametrov. Mišim z induciranim diabetesom smo na hrbtnem delu naredili dve enaki kožni rani s premeroma 5 mm, nato pa jih žrtvovali 2., 4., 7., 10. in 15. dan po ranitvi. Vzorce kontrolne kože in ranjenega tkiva smo analizirali patohistološko, histomorfometrično, imunohistokemijsko in imunokemijsko. V odvzetih vzorcih (tkivni in serumski), smo določali stopnjo regeneracije coriuma (dermisa), izražanje molekul HIF-1? (s hipoksijo induciranega transkripcijskiega dejavnika 1?) in vaskularnega endotelijskega rastnega dejavnika (VEGF) ter koncentracijo kemokina IP-10. Encimsko aktivnost DPP IV/CD26 smo v serumskih in tkivnih vzorcih miši seva C57BL/6, med procesom celjenja ran v pogojih eksperimentalne hiperglikemije, spremljali spektrofotometrično. Ugotovili smo, da so miši CD26-/- v primerjavi z divjim tipom poskusnih živali bolje prenašale tako eksperimentalno hiperglikemijo kot tudi začetno hipoglikemijo. Makroskopske in mikroskopske analize so pri njih pokazale tudi hitrejšo dinamiko celjenja ran. Tako se je pri miših CD26-/- regeneracija coriuma začela prej, vnetna faza procesa celjenja je bila krajša, poleg tega pa je prišlo do hitrejšega nastajanja brazgotin in posledično do njihove hitrejše degradacije. Drugi dan celjenja ran je bilo izražanje HIF-1? pri predstavnikih obeh sevov statistično značilno manjše, 4. dan pa statistično značilno večje (p < 0,05), v primerjavi s kontrolno skupino. Od 4. dneva dalje pa je bila ekspresija HIF-1? v odvzetih tkivih miši CD26-/- statistično značilno višja (p < 0,05) kot v tkivih seva C57BL/6. Ugotovili smo tudi, da je bilo izražanje VEGF pri misših CD26-/- statistično značilno večje (p < 0,05) v vseh testiranih časovnih točah, v primerjavi s poskusnimi živalmi divjega tipa. Serumske koncentracije IP-10 so bile pri miših CD26-/- statistično značilno višje (p < 0,05) v vseh časovnih točkah, v odvzetih vzorcih tkiv pa le na 10. in 15. dan po indukciji kožnih ran. Z analizo rezultatov smo ugotovili tudi, da je bila aktivnost DPP IV/CD26 v serumskih vzorcih miši C57BL/6 statistično značilno manjša (p < 0,05) na 4., v tkivih pa na 2., 4. in 7. dan po indukciji ran v primerjavi s kontrolno skupino. Naši rezultati potrjujejo, da ima molekula DPP IV/CD26 pomembno vlogo pri uravnavanju koncentracije glukoze v krvi. Poleg tega sklepamo, da boljša angiogeneza in večja proliferacija celic v pogojih pomanjkanja DPP IV/CD26 ob povečani lokalni ekspresiji dejavnikov HIF-1?, VEGF in IP-10 potrjujejo, da ima inhibicija DPP IV/CD26 koristne učinke na proces celjenja kožnih ran poskusnih miši, v pogojih eksperimentalno izzvane hiperglikemije. Na podlagi naših izsledkov lahko torej potrdimo velik pomen inhibicije delovanja DPP IV/CD26 v smislu terapevtskega pristopa za zdravljenje sladkorne bolezni in njenih zapletov, še posebej pojava kroničnih ran. Pathophysiological processes of wound healing in hyperglycemia are very complex and insufficiently explained. Dipeptidyl-peptidase IV, or CD26 molecule (DPP IV/CD26) is a multifunctional protein present in a large number of cells in the body and in various biological fluids. Except its involvement in maintenance of glucose homeostasis by regulating the biological activity of various substrates, it also plays an important role in the processes of angiogenesis and proliferation, migration, and apoptosis of various cell types important for the healing of damaged or wounded tissue. Literature data suggest that DPP IV/CD26 inhibition contributes to improved wound healing in patients with diabetes. The aim of the Master's thesis was to establish an experimental model of diabetes (experimental hyperglycemia) in wild-type (C57BL/6) and CD26 deficient (CD26-/-) mice and to determine whether and how in that circumstances, DPP IV/CD26 deficiency influences on the processes of angiogenesis and cutaneous tissue regeneration. The development of diabetes in experimental animals was monitored by clinical and biochemical parameters. Diabetic mice were wounded on the dorsal region (two identical skin wounds with diameters of 5 mm) and sacrificed on days 2, 4, 7, 10, and 15 after wounding. Control skin and wound tissue samples were analyzed pathohistologically, histomorphometrically, immunohistochemically and immunochemically. The degree of corium (dermis) regeneration, the expression of HIF-1α (hypoxia-induced transcription factor 1α) and vascular endothelial growth factor (VEGF) was analysed and the chemokine IP-10 concentration was determined. The enzyme activity of DPP IV/CD26 was determined spectrophotometrically in serum and tissue samples of C57BL/6 mice during the wound healing process in conditions of experimental hyperglycemia. It was found that CD26-/- show improved tolerance of both experimental hyperglycemia and initial hypoglycemia compared to wild type animals. Macroscopic and microscopic analysis showed a faster dynamics of wound healing. Thus, in CD26-/- mice the regeneration of the corium begins earlier, the inflammatory phase of the healing process was shorter accompanied by rapid scar tissue formation and consequently its faster degradation. On the day 2 of wound healing, the expression of HIF-1α was statistically significantly lower in both mice strains and statistically significantly higher on day 4 (p

Natural killer T (NKT) cells are αβ T cell receptor (TCR) expressing innate‐like T cells that display natural killer (NK) cell markers. Based on TCR characteristics, they are divided into two groups restricted to the MHC class I‐like molecule CD1d. Type I NKT cells, most extensively studied, are identified by a semi‐invariant Vα14‐Jα18 (mouse, Vα24‐Jα18 in humans) TCR reactive to the prototypic ligand α‐galactosylceramide presented on CD1d. In contrast, type II NKT cells display diverse TCR reacting to different CD1d‐presented ligands. There are no reagents that identify all type II NKT cells, limiting their exploration. Here, we searched for novel type II NKT cells by comparing Jα18−/−MHCII−/− mice that harbour type II but not type I NKT cells, and CD1d−/−MHCII−/− mice, lacking all NKT cells. We identified significantly larger populations of CD4+ and CD4−CD8− (double negative, DN) TCRβ+ cells expressing NKG2D or NKG2A/C/E in Jα18−/−MHCII−/− mice compared with CD1d−/−MHCII−/− mice, suggesting that 30%‐50% of these cells were type II NKT cells. They expressed CD122, NK1.1, CXCR3 and intermediate/low levels of CD45RB. Further, the CD4+ subset was CD69+, while the DN cells were CD49b+ and CD62L+. Both subsets expressed the NKT cell‐associated promyelocytic leukaemia zinc finger (PLZF) transcription factor and Tbet, while fewer cells expressed RORγt. NKG2D+ CD4+ and DN populations were producers of IFN‐γ, but rarely IL‐4 and IL‐17. Taken together, we identify a novel subset of primary CD4+ and DN type II NKT cells that expresses NKG2 receptors have typical NKT cell phenotypes and a TH1‐like cytokine production.

Цель оценка влияния электромагнитного излучения радиочастотного диапазона на органы кроветворения самок и самцов мышей линии СВА. Методика. Опытные группы животных подвергали воздействию электромагнитного излучения радиочастотного диапазона (диапазон сотовой связи) с интенсивностью 1,2 мВт/см2, периодом экспозиции 10 мин ежедневно в течение 5 сут. Через 30 сут с момента начала облучения часть животных выводили из эксперимента для исследования органов кроветворения, другую часть спаривали для получения потомства. Результаты. Установлено, что воздействие исследуемого фактора вызывает статистически значимые изменения в красном костном мозге, селезенке, тимусе, периферической крови у экспериментальных животных и их потомства. Через 30 сут у облученных самок мышей в крови возрастало количество эритроцитов, у половозрелых самцов количество эритроцитов уменьшалось. Содержание гемоглобина изменялось однонаправленно у облученных самок и самцов, что проявлялось тенденцией к снижению его уровня. У самцов и самок отмечалось снижение числа нейтрофилов. Число лимфоцитов у самцов было в пределах нормы, у самок отмечалась тенденция к увеличению доли лимфоцитов. Однонаправленные изменения у самок и самцов отмечены в эритроидном ростке уменьшение числа полихроматофильных и оксифильных нормоцитов, вследствие ускоренного созревания клеток и их выхода в кровь. В тимусе облученных самок и самцов уменьшалось число ядросодержащих клеток, в селезенке, напротив, их число увеличивалось. У потомства облученных животных снижалось число лейкоцитов с увеличением доли юных и палочкоядерных нейтрофилов, доля лимфоцитов снижалась по сравнению с потомством необлученных животных. Заключение. Выявленные изменения расцениваются как компенсаторноприспособительные реакции в органах кроветворения у экспериментальных животных., The purpose identify the effects of the influence of electromagnetic radiation (EMR) at the radio frequency band hemopoietic organs of males and females CBA mice in different models of irradiation. Methods. The experimental group of animals exposed to RF EMF with an intensity of 1.2 mW / cm2 exposure period of 10 minutes daily for 5 days. After 30 days from the start of irradiation of the animals were taken out of the experiment for the study of blood, the other part mated to produce offspring. Results. It was found that when exposed to the test factor there was a significant change in the bone marrow, spleen, thymus, peripheral blood of experimental animals and their offspring. The findings indicate the development of compensatory and adaptive reactions of the blood of experimental animals., №1 (2019)

Background Numerous studies have explored the anti-tumor effect of berberine (BBR), but little clinical evidence guides the use of BBR in cancer patients. Objectives Our aim was to investigate the impact of BBR on various cancers in healthy animals to promote the transformation from bench to bed. Search methods PubMed, Embase, Springer, and Cochrane databases were searched from January 2000 to October 2018 for relevant articles. Selection criteria Only published studies focusing on the relationship between BBR and various cancers in vivo were qualified. Two review authors independently assessed the risk of bias for each study, and any disagreement was resolved by discussion or by involving a third assessor. Results A total of 26 studies from 2000 to 2018, focusing on various cancer types, including breast cancer, liver cancer, colorectal cancer, nasopharyngeal carcinoma, lung cancer, gastric cancer, neuroepithelial cancer, endometrial carcinoma, esophageal cancer, tongue cancer, cholangiocarcinoma, and sarcoma were included. Overall, BBR reduced tumor volume (SMD =3.72, 95% CI: 2.89, 4.56, Z = 8.73, p

Prvi pisani dokazi korištenja životinja u pokusima potječu još iz Staroga vijeka. Sve do otkrića anestetika životinje su u pokusima proživljavale nezamislive patnje. Prvi akt koji je zabranjivao okrutnost prema životinjama u pokusu donesen je u Britanskom parlamentu 1876. godine. Sredinom dvadesetog stoljeća znanstvenici Russell i Burch u svome su djelu “Načela humanih pokusnih tehnika” postavili temelje nove primijenjene znanstvene discipli- ne koja je trebala poboljšati postupke u radu s laboratorijskim životinjama, a ujedno i pri- donijeti kvaliteti znanstvenog rada u kojem se te životinje koriste. Raščlamba metoda kojima bi se mogli isključiti neljudski postupci prema životinjama u pokusu rezultirala je stvaranjem koncepta 3R načela: Replacement (zamjena), Re- duction (smanjenje) i Refinement (poboljšanje) s ultimativnim ciljem sadržanim u tek jednoj rije- či‒čovječnost. Razvojem znanosti, tehnologije, ali i evolucijom svijesti kako znanstvenika, tako i šire društvene zajednice, 3R načela svakod- nevno se razvijaju, tako da možemo govoriti i o 11R načelima jer se uz tri prethodno navede- na danas još navode i Reliability (pouzdanost), Relevance (pogodnost‒prikladnost), Reproduci- bility (ponovljivost), Rehabilitation (oporavak), Responsibility (odgovornost), Respect (poštova- nje), Redundancy avoidance (izbjegavanje nepo- trebnih ponavljanja) i Regulation (zakonitost postupaka). 3R načelo razvijalo se tijekom pro- teklih pedeset godina osiguravajući okvir za izvođenje humanih pokusnih postupaka pre- ma životinjama, a prvi se puta sam naziv „Na- čela zamjene, smanjenja i poboljšanja“ uvodi u zakonodavstvo Europske unije kroz Direktivu 2010/63/EU. Godine 2010. donesena je Baselska deklaracija kojoj je cilj ujediniti napore znan- stvene zajednice u nastojanjima daljnjeg una- prjeđivanja primjene etičkih principa poput 3R načela u pokusima na životinjama sa svrhom postizanja pozitivnog dijaloga znanstvene za- jednice sa zainteresiranom javnošću. Danas projekt “Odgovorno istraživanje i inovacije” (Responsible Research and Innovation, RRI) Eu- ropskog okvirnog programa Obzor 2020 potiče suradnju i dijalog među istraživačima, građa- nima, političarima, itd. tijekom cijelog procesa istraživanja kako bi se taj proces i njegovi isho- di bolje uskladili s vrijednostima, potrebama i očekivanjima društva., The first written evidence of the use of animals in experiments originated in the Ancient world. Until the discovery of anaesthetics, experimental animals experienced unimaginable suffering. The first act prohibiting cruelty to animals was passed by the British Parliament in 1876. In the mid-20th century, scientists Russell and Burch in their work The Principles of Humane Experimental Technique laid the foundations of a new applied science discipline aimed at improving procedures in working with laboratory animals, while also contributing to the quality of the scientific work in which these animals were used. The analysis of methods to exclude inhumane treatment of animals in experiments resulted in the creation of the concept of the 3R Principle: Replacement, Reduction and Refinement, with the ultimate goal contained in a single word ‒ humaneness. With the development of science, technology, and the evolution of consciousness within the sciences and the general public, the 3R Principle has evolved rapidly, and today has been expanded into the 11R principle, in which Reliability, Relevance Reproducibility, Rehabilitation, Responsibility, Respect, Redundancy avoidance, and Regulation were added to the original 3R principles. Over the past 50 years, the 3R principle has developing into a framework for performing experimental techniques on animals, and for the first time the Principles of Replacement, Reduction and Refinement have been introduced into the European Union legislation through Directive 2010/63/EU. In 2010, the Basel Declaration was adopted, aiming to unite the efforts of the scientific community with the goal for further improvement of the application of ethical principles such as the 3R principles in animal experiments with the aim of achieving positive dialogue between the scientific community and the interested public. Today, the project “European Responsibility Research and Innovation” (RRI) within the European framework programme Horizon 2020 encourages co-operation and dialogue among researchers, citizens, politicians, and others throughout the research process, to better align this process and its outcomes with the values, needs and expectations of society.

В работе приведены результаты изучения некоторых поведенческих показателей у белых беспородных мышей при внутрижелудочном введении смеси, содержащей бензоат натрия, сорбат калия и аскорбиновую кислоту. С помощью метода «открытое поле» регистрировали изменения вертикальной двигательной активности и количество эпизодов груминга, что отражалось в преобладании смешанного тревожно-фобического, пассивно-оборонительного вариантов поведения, агрессии. Пероральное введение смеси консервантов в повышенных дозах в динамике эксперимента оказывает негативное воздействие на поведение животных, что согласуется с нашими данными, полученными ранее., The paper presents the results of studying some behavioral parameters in white outbred mice after intragastric administration of a mixture containing sodium benzoate, potassium sorbate and ascorbic acid. Using the “open field” method, changes in vertical motor activity and the num-ber of grooming episodes were recorded, which was reflected in the prevalence of mixed anxiety-phobic, passive-defensive behaviors, aggression. Oral administration of a mixture of preservatives in high doses in the dynamics of the experiment has a negative impact on the behavior of animals, which is consistent with our data obtained earlier.

В настоящей работе проведено исследование влияния низких температур на интенсивность свободнорадикального окисления липидов и антиоксидантной защиты в тканях внутренних органов (печени, почек, легких, сердца) экспериментальных животных в зависимости от времени экспозиции. Отмечено повышение концентрации низкомолекулярных антиоксидантов в органах животных, время экспозиции которых на холоде длилось 1 ч. Увеличение времени экспозиции крыс на холоде до 3 ч, сопряжено с повышением активности каталазы., The article reports the study on the influence of low temperatures on the intensity of free radical lipid oxidation and antioxidant protection in the tissues of internal organs (liver, kidneys, lungs, heart) in experimental animals, depending on the exposure time. We noted an increase in the concentration of low-molecular antioxidants in the organs of animals, whose exposure time in the cold lasted 1 hour. An increase in exposure time of rats in the cold to 3 hours is associated with an increase in catalase activity., №3(63) (2018)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Após lesões musculares severas, a comunicação entre os macrófagos (Mϕs) e células miogênicas é essencial para induzir o reestabelecimento das estruturas que compõem o músculo esquelético. No entanto, a comunicação e contribuição dos Mϕs após lesão induzida pelo exercício físico (LIEF) ainda é pouco compreendida. O objetivo deste estudo foi avaliar a participação de dois subtipos de Mϕs no músculo esquelético após LIEF. Os camundongos foram expostos ao protocolo de exercício em downhill (18 tiros de corrida, cada tiro composto de 5 min por 2 min de recuperação passiva, a inclinação da descida foi fixada a -16°). A eutanásia ocorreu imediatamente, 24, 48 e 72 horas pós-exercício (CTRL, D1, D2 e D3 respectivamente). O músculo tríceps braquial (TB), gastrocnêmio (GAST), quadríceps (QUAD) e tibial anterior (TA) foram dissecados e submetidos a extração proteica, montagem histológica ou digeridos para as análises de citometria de fluxo. A marcação por anticorpo F4/80 foi utilizada para quantificar Mϕs nos músculos. Os anticorpos CD45, CD64 e Ly6C foram utilizados para determinar os diferentes subtipos de Mϕs. Ainda, foi realizado análise por multiplex e western blot para a quantificação da expressão proteica e de algumas citocinas. Não foram observadas diferenças estatísticas na quantidade de células positivas para F4/80 entre os músculos analisados. No entanto, por meio de citometria, observamos aumento de células positivas para CD45 (p < 0,05), e de Ly6C+ (p< 0,05) três dias após o exercício. Neste mesmo momento também observamos tendência de redução de Ly6C- (p = 0,07), aumento na concentração de citocinas pró inflamatórias (IL-1β, TNF-α, MCP-1 e IFN-γ, todas com p < 0.05) e correlação inversa com os entre Ly6C- e Ly6C+ (p < 0,05 e C.C = 0,81). Por fim, também foi observado uma tendência na diminuição de IL-4 (p = 0,07) simultaneamente ao aumento de IL-13). Coletivamente, nossos resultados sugerem que o protocolo downhill foi responsável por induzir uma inflamação e que esta inflamação foi responsável por recrutar e modular os fenótipos de Mϕs no músculo TB. Ainda, a secreção de algumas citocinas como a TNF-α e a IL-13 possivelmente podem estar associadas com os processos de miogênese após o protocolo de LIEF. After skeletal muscle damage, the communication between inflammatory macrophages (Mϕs), myogenic cells by cytokines secretion are essential for re-establishment of skeletal muscle structure. Although, these communications are poorly exploring after exercise induce muscle damage (EIMD). The aim of this study is to quantify Mϕ subtypes within mice muscles after downhill exercise protocol. Methods: Mice were exposed to an intermittent protocol of downhill exercise (18 bouts of running, each bout was consisted by 5 minutes with 2 minutes of rest interval. The treadmill inclination was set at -16°) and euthanized before (CTRL), one (D1), two (D2) and three (D3) days after exercise. After euthanasia, the Triceps brachial (TB), gastrocnemius (GAST), quadriceps (QUAD) and Tibialis anterior (TA) were collected, and submitted to protein extraction, immunostaining procedures or flow cytometry analysis. The anti-F4/80+ positive cells were utilized to determine accumulation of Mϕ into muscles and the CD45, CD64 and Ly6C antibodies were used to quantify Mϕ subsets in TB. Additionally, multiplex analysis was use for muscle cytokines quantification. Results: We did not observe any statistical differences on F4/80+ positive cells among muscles analyzed. By flow cytometry analysis, we observed an increase of CD45+ (P< 0.05) cells at day 3, with similar increase of Ly6C+ Mϕs (P< 0.05), but the Ly6C- Mϕs trended to a slight decrease (P= 0.07). We also noticed an elevation of the pro-inflammatory cytokines (IL-1β, TNF-α, MCP-1 and IFN-γ all with P< 0.05) along with an inverse correlation (P< 0.05 e C.C=-0.81) between Ly6C+ and Ly6C- Mϕs, which suggest that this inflammation resulted Mϕ subset change on TB muscles 3 days after downhill exercise. Nevertheless, we also observed an increase we also observed a decrease trend of IL-4 (P= 0.07) and a significant increase of IL-13 (P< 0.05).Collectively, ours results indicates that downhill exercise induces an inflammation which were associate with Mϕs subset change. Moreover, the secretion of some cytokines also indicates that these molecules might have an important role on myogenesis process after EIMD. FAPESP: 2014/04504-6 FAPESP: 2016/06817-7 Dados abertos - Sucupira - Teses e dissertações (2018)

Rabbit mesenchymal stem cells (MSCs) are important seed cells in regenerative medicine research, particularly in translational research. In the current study, we showed that rabbit subchondral bone is a reliable source of MSCs. First, we harvested subchondral bone (SCB) from the rabbit knee-joint and initiated the MSC culture by cultivating enzyme-treated SCB. Adherent fibroblast-like cells that outgrew from SCB fulfill the common immuno-phenotypic criteria for defining MSCs, but with low contamination of CD45+ hematopoietic cells. Interestingly, differentiated SCB-MSCs expressed osteogenic and chondrogenic markers at significantly higher levels than those in bone marrow cell suspension-derived MSCs (BMS-MSCs) (P0.05) were observed. Moreover, the results of the colony forming unit-fibroblast assay and sphere formation assay demonstrated that the SCB-MSCs had increased self-renewal potential. SCB-MSCs expressed higher levels of the stemness markers Nanog, OCT4, and Sox-2 compared to in BMS-MSCs (P

Laboratory is the main supporting to reach the learning aim that appropriate competency for Nutrition Department students. Utilization of experimental animals as objects experiment in the laboratory to test in vivo ( laboratory with experimental animals ) not available yet, this laboratory is important to development of science related with diet, chemical compounds that has a positive role to health. Laboratory of experimental animals is not available yet, therefore necessary to design the laboratory of experimental animals. Purposes of Research : to design laboratory management of experimental animals, management of experimental animals cages and management of experimental animals waste. Design of research includes descriptive qualitative with comparation study in laboratory of experimental animals at Universitas Gajah Mada ( UGM ) and Universitas Negeri Semarang ( UNNES ). Data collection with literature study, interview, visit to laboratory of experimental animals. Results of research : hamster, white mouse and rabbit are used as experiment because of the fisiologis excuses, these animals are able to be simulation model because have some similiars with the condition of human body. Animal source must be from a certified institute that provides experimental animals who is able to guarantee animals health. Original genetic and homogenous. Cage in accordance with necessaries. There are three types : cage enlargement, cage for feeding and mating. Cage must be kept clean. Diet should be made periodically as needed. Management staff who has biology educational background one or two person and two other staff. Laboratory design of experimental animals at Nutrition Department has just been developed as simple applied laboratory, without reproduction management and enlargement of experimental animals. Laboratory management of experimental animals refers to the current system. Design of cage has a size 4m x 4m which is equipped sanitation facilities / sink and prep table. Conclusion : Hamster and white mice are used for experimental animals. The cage treatment and cage maintenance are cage design that is required. One management staff with the biology educational backgroud is required and empowering an existing. Room design 4m x 4m which is equipped a sink and prep table for equipment.

We assessed the level of pro- (IL1β, FNOα, IL2) and anti-inflammatory (IL10) cytokines in the anterior chamber fluid of experimental animals on the 3rd, 7th and 30th day, after an experimental ciliary body destruсtion with porous titanium nickelid cryoapplicator. Consistent changes of the cytokine content at the local level are reflecting the reactivity of the experimental animals as an answer both to surgery in ciliary body area, and to its experimental cyclodestruction using the cryoapplicator from titanium nickelid. The predominance of pro-inflammatory cytokines’ potential was found during the early period (3-7 day), and the balance normalization of pro- and anti-inflammatory cytokines was found at late observation period (30 days) due to the increase of the anti-inflammatory IL-10 production; this effect was maintained at the end of the long-term study period (1.5 years).

Changes in the composition of intestinal microbiota, namely the reduction of microorganisms capable of metabolizing oxalates, is one of the main factors in the development of hyperoxaluria. The purpose of this study was to determine the quantity of oxalate-degrading bacteria in fecal biopsy of different species of laboratory animals and to investigate the relationship between the amount of oxalate in the diet and the level of oxalate-degrading bacteria. Methods. The object of the study was the content of oxalate-degrading bacteria in fecal biopsy of experimental animals: nonbreeding rats (n = 12); Wistar rats (n = 12); mice Balb C line (n =12); сhinchilla rabbits (n= 10). The quantity of oxalate-degrading bacteria was determined by culture method on a highly selective Oxalate Medium. Results. Current data shows that the content of oxalate-degrading bacteria in fecal biopsy depends on the species of the animals and the high content of oxalates in the diet. In Wistar line rats, oxalate-degrading bacteria were found in 100 % of the animals, in non-breeding rats – 58 %, in mice Balb C line – 42 %, in сhinchilla rabbits – 7 %. The highest quantity of oxalatedegrading bacteria in grams of fecal biopsy was found in Wistar rats – lg 6,12 ± 0,63 CFU/g, in non-breeding rats – lg 2,97 ± 0,34 CFU/g, in mice lg 2,4 ± 0,41 CFU/g. The least quantity of oxalate-degrading bacteria was detected in rabbits (lg 2,1 ± 0,5 CFU/g). A 14-day administration of Na2C2O4 to non-breeding rats has led to an increase in the quantity of oxalate-degrading bacteria in fecal biopsy. During the first seven days of the high-oxalate diet 57 % animals had increased the quantity of oxalate-degrading bacteria in a gram of feces. After 14 days, the further increase of oxalatedegrading bacteria in a gram of feces was observed in 25 % animals. In general, the quantity of oxalate-degrading bacteria during high-oxalate diet has increased by two orders. Conclusions. Animals with non-detectable level of oxalate-degrading bacteria in feces might be attributed to the potential risk group of hyperoxaluria and the formation of oxalate stones.

The beginnings of research on animals date back to ancient precursors of science and medicine. In the history of vivisection the most significant were representatives of 19th century French science such as physiologist Francois Magendie who became famous with the public exposure of dogs nervous system by nailing them to the tables. Such experiments were always excused by philosophers: Descartes, Kant and others. Now, thanks to new legal solutions animal experimentation is more restricted and controlled. This text is an attempt to systematize the history of experiments on animals and to confront them with the present state of animal protection used for experiments.

Оценивали влияние низких температур (-10±2 0С на процессы перекисного окисления липидов в тканях внутренних органов (печени, почках, легких, сердца) крыс. Опытная и контрольная группы включали по 8 крыс линии Вистар. Животных опытной группы экспонировали нахолоде в течение 14 дней по 1 ч. На холоде температура лапок, измеренная электротермометром с игольчатымдатчиком, понижалась на 6,8 0С до (+20,5±0,5) 0С, хвоста на 3,7 0С до (+21,4±0,7) 0С, по сравнению с контролем. Это свидетельствует о нарушении микроциркуляции и возникновении гипоксии в конечностях, следствие интенсификация перекисных процессов. В конце опыта отбирали по 100 мг ткани печени, почек, легкого и сердца, которые исследовали с помощью спектрофотометра SPECORD 40. У подопытных животных концентрация малонового диальдегида конечного продукта перекисного окисления липидов, была значительно ниже контрольных значений: в печени в 1,6 раза (6,64 нмоль/г), почках -в 1,7 раза (9,74 нмоль/г), легких в 3,0 раза (5,02 нмоль/г). Уровень диеновых коньюгатов первичных продуктов перекисного окисления липидов был значительно выше, в сравнении с интактной группой, в печени в 1,7раза (3,79 мкмоль/л), почках-в 5,6раз (5,55 мкмоль/л), легких-в 1,4 раза (7,10 мкмоль/л), сердце в 1,7 раза (4,51 мкмоль/л). Суммарное содержание низкомолекулярных антиоксидантов было достоверно выше, чем в контроле, в печени в 1,8 раз (29,78 мг-экв кверцетина/г), почках-в 2,0раза (76,57мг-экв кверцетина/г), легких-в 22,0 раза (44,18 мг-экв кверцетина/г). Активность каталазы была существенно ниже контрольных значений в печени в 1,4 раза (15,14 мккат/г), почках в 1,7раза (10,45 мккат/г), легких в 1,7раза (9,89 мккат/г), сердце в 1,5 раза (8,56 мккат/г)., The authors determined the influence of low temperatures (-10 ± 2) C on the processes of lipid peroxidation in tissues of viscera (liver, kidneys, lungs, heart) of rats. An experimental and control group consisted of eight Wistar rats. Animals from the experimental group were exposed to the cold temperature for 14 days. In cold, the temperature of paws, measured by an electric thermometer with a needle sensor, decreased by 6.8 C to (+20.5 ± 0.5) C and of the tail by 3.7 C to (+21.4 ± 0.7) C, in comparison with the control. It indicates a violation of microcirculation and occurrence of hypoxia in the limbs and, as a result, intensification of peroxide processes. At the end of the experiment, 100 mg of liver, kidney, lung and heart tissue were sampled, which were examined using SPECORD 40 spectrophotometer. In the experimental animals, the concentration of malondialdehyde, the end product of lipid peroxidation, was significantly lower than control values: 1.6 times in the liver tissues (6.64 nmol/g), 1.7 times in kidneys (9.74 nmol/g), 3.0 times in lungs (5.02 nmol/g). The level of diene conjugates, primary products of lipid peroxidation, was reliably higher, in comparison with the intact group, in liver tissues 1.7 times (3.79 micromol/L), kidneys 5.6 times (5.55 micromol/L), lungs 1.4 times (7.10 micromol/L), myocardium 1.7 times (4.51 micromol/L). The total content of low-molecular antioxidants was reliably higher, in comparison with the control group, in the liver tissues 1.8 times (29.78 mg-eq/g of quercetine), kidneys 2.0 times (76.57 mg-eq/g of quercetine), lungs 22.0 times (44.18 mg-eq/g of quercetine). The catalase activity was significantly lower than the control values: in the liver tissue 1.4 times (15.14 microkat/g), in kidneys 1.7 times (10.45 microkat/g), in lung 1.7 times (9.89 microkat/g), in myocardium 1.5 times (8.56 microkat/g).

Topicality. Insufficient light in the world scientific literature the problem of changing the regulatory mechanisms of the immune response depending on age. Little-studied molecular events and cellular interactions underlying the mechanisms of immunoreactivity for the development of adaptation at different stages of ontogenesis.Aim. To study peculiarities of changes of phagocytic activity of neutrophils and evaluation of their enzymatic activity in the experimental animals of different ages on the background of inflammatory process and the use of immunocorrectjrs polioksidonium and mixfactor.Materials amd methods. In the course of work conducted the study of the cellular component of innate immunity in experimental animals of different ages on the background of the introduction of a suspension of E. coli and the use of immunomodulators at different stages of the experiment by microscopic method.Results and discussion. The study revealed that polioksidonium should be applied after the activation of the inflammatory response since this drug has a pronounced antioxidant activity, which is negative to ensure the phagocytosis of neutrophils through reactive oxygen species, which provides endotsitoz bacterial antigens.Conclusions. The introduction of an experimental composite preparation mixfactor resulted in stimulation of phagocytosis activity, which is evidence about adaptive interaction processes of reactivity and resistance of experimental animals of different age., Актуальность. Недостаточно освещена в мировой научной литературе проблема изменения регуляторных механизмов иммунного ответа в зависимости от возраста. Мало изучены молекулярные события и клеточные взаимодействия, лежащие в основе механизмов иммунореактивности, обеспечивающие развитие адаптации на разных этапах онтогенеза.Целью нашей работы было изучение особенностей изменения фагоцитирующей активности нейтрофилов и оценка их ферментативной активности у экспериментальных животных разного возраста на фоне воспалительного процесса и применение иммунокорректоров полиоксидония и миксфактора.Материалы и методы. В ходе работы проводились исследования клеточного звена врожденного иммунитета у экспериментальных животных разного возраста на фоне введения взвеси E. сoli и применения иммунокорректоров на разных этапах эксперимента с помощью микроскопического метода.Результаты и их обсуждение. В ходе исследования выявлено, что полиоксидоний следует применять после активации воспалительной реакции, так как этот препарат имеет выраженную антиоксидантную активность, что является негативным для обеспечения фагоцитоза нейтрофилов с помощью активных форм кислорода, который обеспечивает эндоцитоз бактериальных антигенов.Выводы. Введение экспериментального композитного препарата миксфактора привело к стимуляции активности фагоцитоза, что свидетельствует об адаптивном взаимодействии процессов реактивности и резистентности экспериментальных животных разного возраста., Актуальність. Недостатньо висвітлена в світовій науковій літературі проблема зміни регуляторних механізмів імунної відповіді в залежності від віку. Мало вивчені молекулярні події та клітинні взаємодії, що лежать в основі механізмів імунореактивності, які забезпечують розвиток адаптації на різних етапах онтогенезу.Метою нашої роботи було вивчення особливостей зміни фагоцитуючої активності нейтрофілів та оцінка їх ферментативної активності в експериментальних тварин різного віку на тлі запального процесу і застосування імунокореторів поліоксидонію та міксфактора.Матеріали та методи. Під час роботи проводились дослідження клітинної ланки вродженого імунітету в експериментальних тварин різного віку на тлі введення суспензії Е. coli та застосування імунокоректорів на різних етапах експерименту за допомогою мікроскопічного методу.Результати та їх обговорення. В результаті дослідження виявлено, що поліоксидоній слід застосовувати після активації запальної реакції, так як цей препарат має виражену антиоксидантну активність, що є негативним для забезпечення фагоцитозу нейтрофілів за допомогою активних форм кисню, що забезпечує ендоцитоз бактеріальних антигенів.Висновки. Введення експериментального композитного препарату міксфактора призвело до стимуляції активності фагоцитозу, що свідчить про адаптивний зв’язок процесів реактивності і резистентності експериментальних тварин різного віку.