Administration of vitamin D3 improves antimetastatic efficacy of cancer vaccine therapy of Lewis lung carcinoma

Aim: To analyze antitumor efficacy of experimental cancer vaccine therapy combined with introduction of vitamin D3 (VD3) for treatment of Lewis lung carcinoma (3LL). Materials and Methods: Cancer vaccines composed from recombinant murine beta-defensin-2 (mBD-2) and 3LL cell lysate, or DNA, coding for mBD-2-Muc1 fusion construct cloned in pcDNA3+ vector, were prepared and used for intradermal vaccination. Experimental cancer vaccines introduced i. d. at therapeutic and prophylactic regimens to 3LLbearing C57Bl mice, were applied alone or in combination with VD3 (administered per os) and/or low-dose cyclophosphamide (CP, administered intraperitoneal). Efficacy of treatments was analyzed by primary tumor growth dynamics indexes and by metastasis rate in vaccinated animals. Results: As it has been shown, administration of the protein-based vaccine composed from mBD-2 and 3LL cell lysate in combination with VD3 and CP, but not in VD3 free setting, led to significant suppression of primary tumor growth (p < 0.005) and had significant antimetastatic effect. Introduction of VD3 with or without CP in the scheme of treatment with mBD-2-Muc1-DNA vaccine at therapeutic regimen has led to significant suppression of primary tumor (p < 0.05) and metastasis volumes (p < 0.005), while in the groups of animals treated with DNA-vaccine + VD3 with or without CP at prophylactic regimen, significant antimetastatic effect (p < 0.05) and elevation of average life-span (p < 0.05) have been registered. Conclusion: The results of this pilot study have shown promising clinical effects of VD3 administration in combination with cancer vaccinotherapy in vivo

Human beta-defensin-2 controlS cell cycle in malignant epithelial cells: in vitro study

Aim: In the present research we analyze the mechanism of human beta-defensin-2 (hBD-2) influence on cultured malignant epithelial cell growth. Materials and Methods: The analysis of a concentration-dependent effect of recombinant hBD-2 (rec-hBD-2) on cell growth patterns and cell cycle distribution has been performed in vitro with 2 cell lines (human lung adenocarcinoma A549 cells and human epidermoid carcinoma A431 cells) using MTT test, flow cytometry and direct cell counting. To study intracellular localization of hBD-2 immunocytofluorescent and immunocytochemical analyses were applied, and effect of hBD-2 on signal cascades involved in cell cycle regulation has been studied by Western blotting. Results: According to our data, rec-hBD-2 exerts a concentration-dependent effect on the viability of cultured A549 and A431 cells. It causes proproliferative effect at concentrations below 1 nM, significant suppression of cell proliferation at concentration range from 10 nM to 1 µM (p<0.05), and cell death at higher concentrations. Using flow cytometry we have demonstrated that hBD-2 dependent growth suppression is realized via cell cycle arrest at G1/S phase (p<0.05). Also, we have registered significant activation of pRB and decreased expression of Cyclin D1 in cells treated with the defensin compared to untreated control cells, while the expression of p53 remains unaffected. The study of intracellular localization of hBD-2 in these cells has revealed that exogeneously added defensin molecules enter the cells, are distributed throughout the cytoplasm and could be detected in cell nuclei. The model study using A549 cells treated with 1,25-(OH)2D3 has shown similar cell growth suppression effect of native endogenously produced hBD-2. Conclusion: The results of our study suggest that in malignant epithelial cells hBD-2 may control cell growth via arrest of G1/S transition and activation of pRB

Polymorphisms of the GCLC Gene Are Novel Genetic Markers for Susceptibility to Psoriasis Associated with Alcohol Abuse and Cigarette Smoking

The aim of this pilot study was to investigate whether single nucleotide polymorphisms in the gene encoding the catalytic subunit of glutamate cysteine ligase are associated with the risk and clinical features of psoriasi

Immunohistochemical analysis of beta-defensin-2 expression in human lung tumors

The aim of this paper is to present research was directed on analysis of the expression patterns of human beta-defensin-2 (hBD-2) in human lung tumors

Polysemy of greek suffix -os- in the formation of derivative clinical terms

The aim of the study - to analyze the meanings of the suffix -os- in the formation of derivative clinical terms.Цель исследования - анализ значений суффикса -os- при формировании производных клинических термино

ФИЗИОЛОГИЧЕСКИЙ ЭФФЕКТ ПРОБИОТИКА ЗООВЕСТИНА В ЭКСПЕРИМЕНТАХ НА НЕПРОДУКТИВНЫХ ЖИВОТНЫХ

The article focuses on the fact, that probiotic specimens affect insufficiently due to lack of specific microorganisms for gut colonization, response to antibiotics or some feeds, way of applying and doses and periods of applying. The authors explain the importance of estimating the effect of applying new probiotics on domestic animals. The researchers conducted experiments on cats and dogs; the experimental results show the effect of probiotic Zoovestin. The paper estimates the state of blood in the beginning of the experiment and declares that blood of cats and dogs corresponds to the standards. The authors observed the microorganisms as coccus, coliform bacillus with low enzyme ability, yeast-like fungi, globulolytic coliform bacillus and proteus in gut microflora of pets. There were no changes in blood indexes of pets observed. Zoovestin improved gut flora of cats and dogs and made its indexescorrepond to the physiological standards. The researchers observed an increase in number of bifid bacterium and coliform bacillus with low enzyme ability. The number of coccus flora and coliform bacillus with low enzyme ability was reduced and globulolytic coliform bacillus and proteus were removed. The researchers observed better appetite and exterior that characterizes their welfare. Пробиотические препараты иногда дают недостаточный эффект, что связывается с необходимостью специфических типов микроорганизмов для колонизации кишечника определенного вида животных, чувствительностью к антибиотикам или некоторым кормам, способом применения, включая дозы и периоды использования. В связи с этим особую актуальность приобретает оценка физиологического эффекта на организм животных новых пробиотических препаратов. В опытах на кошках и собаках показано действие пробиотика на основе бифидобактерий – зоовестина. Оценка гематологического статуса в начале эксперимента показала, что и кошки, и собаки характеризуются показателями крови в границах нормы. В то же время в кишечной микрофлоре были в значительной степени представлены такие микроорганизмы, как кокки, кишечная палочка с низкой ферментативной активностью, дрожжеподобные грибы, гемолитическая кишечная палочка и протей. Гематологические показатели животных в процессе исследований значительных изменений не претерпели. Зоовестин стабилизировал кишечную флору кошек и собак и привел ее показатели к физиологическим нормам. Произошло некоторое повышение количества бифидобактерий и кишечной палочки с нормальной ферментативной активностью. Снизилось до нормы количество кокковой флоры и кишечной палочки с низкой ферментативной активностью, элиминирована гемолитическая кишечная палочка и протей. Улучшился аппетит и внешний вид животных, что характеризует их благополучие

Expression of the NH2-Terminal Fragment of RasGAP in Pancreatic β-Cells Increases Their Resistance to Stresses and Protects Mice From Diabetes

OBJECTIVE: Our laboratory has previously established in vitro that a caspase-generated RasGAP NH(2)-terminal moiety, called fragment N, potently protects cells, including insulinomas, from apoptotic stress. We aimed to determine whether fragment N can increase the resistance of pancreatic beta-cells in a physiological setting. RESEARCH DESIGN AND METHODS: A mouse line, called rat insulin promoter (RIP)-N, was generated that bears a transgene containing the rat insulin promoter followed by the cDNA-encoding fragment N. The histology, functionality, and resistance to stress of RIP-N islets were then assessed. RESULTS: Pancreatic beta-cells of RIP-N mice express fragment N, activate Akt, and block nuclear factor kappaB activity without affecting islet cell proliferation or the morphology and cellular composition of islets. Intraperitoneal glucose tolerance tests revealed that RIP-N mice control their glycemia similarly as wild-type mice throughout their lifespan. Moreover, islets isolated from RIP-N mice showed normal glucose-induced insulin secretory capacities. They, however, displayed increased resistance to apoptosis induced by a series of stresses including inflammatory cytokines, fatty acids, and hyperglycemia. RIP-N mice were also protected from multiple low-dose streptozotocin-induced diabetes, and this was associated with reduced in vivo beta-cell apoptosis. CONCLUSIONS: Fragment N efficiently increases the overall resistance of beta-cells to noxious stimuli without interfering with the physiological functions of the cells. Fragment N and the pathway it regulates represent, therefore, a potential target for the development of antidiabetes tools

Transgenic Overexpression of Active Calcineurin in β-Cells Results in Decreased β-Cell Mass and Hyperglycemia

BACKGROUND:Glucose modulates beta-cell mass and function through an initial depolarization and Ca(2+) influx, which then triggers a number of growth regulating signaling pathways. One of the most important downstream effectors in Ca(2+) signaling is the calcium/Calmodulin activated serine threonine phosphatase, calcineurin. Recent evidence suggests that calcineurin/NFAT is essential for beta-cell proliferation, and that in its absence loss of beta-cells results in diabetes. We hypothesized that in contrast, activation of calcineurin might result in expansion of beta-cell mass and resistance to diabetes. METHODOLOGY/PRINCIPAL FINDINGS:To determine the role of activation of calcineurin signaling in the regulation of pancreatic beta-cell mass and proliferation, we created mice that expressed a constitutively active form of calcineurin under the insulin gene promoter (caCn(RIP)). To our surprise, these mice exhibited glucose intolerance. In vitro studies demonstrated that while the second phase of Insulin secretion is enhanced, the overall insulin secretory response was conserved. Islet morphometric studies demonstrated decreased beta-cell mass suggesting that this was a major component responsible for altered Insulin secretion and glucose intolerance in caCn(RIP) mice. The reduced beta-cell mass was accompanied by decreased proliferation and enhanced apoptosis. CONCLUSIONS:Our studies identify calcineurin as an important factor in controlling glucose homeostasis and indicate that chronic depolarization leading to increased calcineurin activity may contribute, along with other genetic and environmental factors, to beta-cell dysfunction and diabetes

Association of hypoxia inducible factor-1 alpha gene polymorphism with both type 1 and type 2 diabetes in a Caucasian (Hungarian) sample

BACKGROUND: Hypoxia inducible factor-1 alpha (HIF-1alpha) is a transcription factor that plays an important role in neo-vascularisation, embryonic pancreas beta-cell mass development, and beta cell protection. Recently a non synonymous single nucleotide polymorphism (g.C45035T SNP, rs11549465) of HIF-1alpha gene, resulting in the p.P582S amino acid change has been shown to be associated with type 2 diabetes (T2DM) in a Japanese population. Our aim was to replicate these findings on a Caucasian (Hungarian) population, as well as to study whether this genetic effect is restricted to T2DM or can be expanded to diabetes in general. METHODS: A large Caucasian sample (N = 890) was recruited including 370 T2DM, 166 T1DM and 354 healthy subjects. Genotyping was validated by two independent methods: a restriction fragment analysis (RFLP) and a real time PCR using TaqMan probes. An overestimation of heterozygotes by RFLP was observed as a consequence of a nearby SNP (rs34005929). Therefore genotyping results of the justified TaqMan system were accepted. The measured genotype distribution corresponded to Hardy-Weinberg equilibrium (P = 0.740) RESULTS: As the TT genotype was extremely rare in the population (0.6% in clinical sample and 2.5% in controls), the genotypes were grouped as T absent (CC) and T present (CT and TT). Genotype-wise analysis showed a significant increase of T present group in controls (24.0%) as compared to patients (16.8%, P = 0.008). This genetic effect was demonstrated in the separated samples of type 1 (15.1%, P = 0.020), and also in type 2 (17.6%, P = 0.032) diabetes. Allele-wise analysis gave identical results showing a higher frequency of the T allele in the control sample (13.3%) than in the clinical sample (8.7%, P = 0.002) with similar results in type 1 (7.8%, P = 0.010) and type 2 (9.1%, P = 0.011) diabetes. The odds ratio for diabetes (either type 1 or 2) was 1.56 in the presence of the C allele. CONCLUSION: We confirmed the protective effect of a rare genetic variant of HIF-1alpha gene against type 2 diabetes in a Caucasian sample. Moreover we demonstrated a genetic contribution of the same polymorphism in type 1 diabetes as well, supporting a possible overlap in pathomechanism for T2DM and a T1DM

Relationship of depressive disorders with hypertension, its control and other metabolic risk factors in the Tyumen Oblast population of men and women. Data from the study “Epidemiology of Cardiovascular Diseases and their Risk Factors in Regions of Russian Federation” (ESSE-RF)

Aim. To study the association between depression and metabolic cardiovascular risk factors, hypertension (HTN) and its control in a random sample of Tyumen Oblast population of men and women aged 25-64 years.Material and methods. The study object was a random sample of the population of the Tyumen and the Tyumen Oblast aged 25-64 years, examined as part of the ESSE-RF epidemiological study. The study included 1658 participants. Among them, 30,3% (n=503) were men, while 69,7% (n=1155) — women. Mean age was 48,9±11,4 years. The prevalence of metabolic risk factors (hyperlipidemia, carbohydrate metabolism disorder, obesity), hypertension and the likelihood of its control in men and women with different levels of depressive disorders diagnosed using the HADS scale were assessed.Results. Compared with participants without depression, persons with psychological disorders were significantly more likely to have HTN (55,5% vs 47,6%, p&lt;0,01), elevated levels of total cholesterol (TC) (63,9% vs 54,0%, p&lt;0,01) and low-density lipoproteins (LDL) (66,7% vs 60,3%, p&lt;0,05), carbohydrate metabolism disorders (8,3% vs 5,2% p&lt;0,05), obesity (49,2% vs 37,7%, p&lt;0,01). Significantly more often hypertensive subjects without depression took antihypertensive drugs effectively (odds ratio (OR) — 1,747, 95% confidence interval (CI), 1,001-3,053) and controlled blood pressure (OR — 1,533, 95% CI, 1,05-2,36). There was no association between the use of antihypertensives and the level of depressive disorders. Among women with depression (HADS&gt;7), dyslipidemia (65,5% vs 57,4% for TC, p&lt;0,05; 71,0% vs 62,9% for LDL, p&lt;0,05), carbohydrate metabolism disorders (10,1% vs 5,2%, p&lt;0,01), obesity (53,3% vs 43,2%, p&lt;0,01), HTN (60,6% vs 45,6%, p&lt;0,01) were more common. Men with clinical depression were more likely to have HTN (69,0% vs 47,7%, p&lt;0,05), with a high level of depression — hyperlipidemia (58,9% vs 46,7% for TC, p&lt;0,05; 67,1% vs 53,9% for LDL, p&lt;0,05). Women with elevated depression levels were less likely to take antihypertensive drugs (30% vs 49,4%, p&lt;0,01) and control hypertension (13,8% vs 21,2%, p&lt;0,05).Conclusion. The data obtained confirm the association of depressive disorders with metabolic risk factors and the likelihood of HTN control, which is especially significant among women