Intragenic MicroRNAs Binding Sites in MRNAs of Genes Involved in Carcinogenesis

MiRNAs participate in gene regulation of translation. Some studies have investigated the interactions between genes and intragenic miRNAs. It is important to study the miRNA binding sites of genes involved in carcinogenesis. RNAHybrid 2.1 and ERNAhybrid programmes were used to compute the hybridization free energy of miRNA binding sites. Of these 54 mRNAs, 22.6%, 37.7%, and 39.7% of miRNA binding sites were present in the 5'UTRs, CDSs, and 3'UTRs, respectively. The density of the binding sites for miRNAs in the 5'UTR ranged from 1.6 to 43.2 times and from 1.8 to 8.0 times greater than in the CDS and 3'UTR, respectively. Three types of miRNA interactions with mRNAs have been revealed: 5'- dominant canonical, 3'-compensatory, and complementary binding sites. MiRNAs regulate gene expression, and information on the interactions between miRNAs and mRNAs could be useful in molecular medicine. We recommend that newly described sites undergo validation by experimental investigation.

Pyrite from Zones of Mz-Kz Reactivation of Large Faults on the Eastern Slope of the Ural Mountains, Russia

Pyritisation halos are identified in weathering crusts and unconsolidated formations at five locations within large fault structure of the Urals’ eastern slope. Electron microscopy reveals the presence of inclusions and growths on pyrite faces – normally on cubic pyrite with striations, or combinations of cubes and other forms. Following neogenesis types are established: native elements and intermetallic compounds (including gold and silver), halogenides, sulphides, sulfosalts, tellurides, sulphotellurides, selenides, tungstates, sulphates, phosphates, carbon-based substances. Direct relationship is noted between amount and diversity of such mineral phases, and proximity to and scale of ore-grade mineralization. Gold and silver, both in native form and within tellurides, presence of lead (galena, native lead), native tungsten, and, possibly, molybdenite and sulfosalts can indicate gold-bearing formations. First find of native tungsten in the Urals is for the first time – in crystallised and druse-like form. Link is suggested between unusual mineralization and “reducing” hydrothermal fluids from deep-seated faults at later stages of Urals’ reactivation. 

Changes in Selected Fuel Properties of Sewage Sludge as a Result of its Storage

The article presents test results on the changes occurring in sewage sludge during the process of its storage. Tests were conducted on mechanically dehydrated sewage sludge derived from large municipal sewage treatment plants equipped with biological sewage treatment systems. In testing presented in the paper the focus was on the basic fuel properties of sewage sludge: moisture content, heat of combustion, carbon share. In the first part of the article the overview of the issues concerning the sewage sludge management is presented and the genesis of tests is explained. Further in the paper, selected results of conducted tests are discussed. Changes in tested parameters were determined in the period of a 10- month sewage storage.

Histogenesis of Rabbit Vallate Papillae

The gustatory system allows animals to distinguish varieties of food and affects greatly the consumption of food, hence the health and growth of animals. In the current study, we investigated the histogenesis of vallate papillae (VLP) in the rabbit tongue using light and scanning electron microscopy. Samples were obtained from rabbit embryos at the embryonic days 16-30 (E16-30), and from newborns until maturity; 6 months. At E16, the first primordia of vallate papillae were observed as small pits on the surface epithelium of the tongue-s root. At E18, the caudal part was prominent with loose mesenchymal tissue core; meanwhile the rostral part of the papilla was remained as a thick mass of epithelial cells. At E20-24, the side epithelium formed the primitive annular groove. At E26, the primitive taste buds appeared only at the papillary surface and reached their maturity by E28. The annular groove started to appear at E26 became more defined at E28. The definitive vallate papillae with substantial number of apparently mature taste buds were observed by the end of the second week. We conclude that the vallate papillae develop early and mature during the early postnatal life.

Phenotypes of B Cells Differ in EBV-positive Burkitt-s lymphoma Derived Cell Lines

Epstein-Barr virus (EBV) is implicated in the pathogenesis of the endemic Burkitt-s lymphoma (BL). The EBVpositive BL-derived cell lines initially maintain the original tumor phenotype of EBV infection (latency I, LatI), but most of them drift toward a lymphoblast phenotype of EBV latency III (LatIII) during in vitro culturing. The aim of the present work was to characterize the B-cell subsets in EBV-positive BL cell lines and to verify whether a particular cell subset correlates with the type of EBV infection. The phenotype analysis of two EBV-negative and eleven EBV-positive (three of LatI and eight of LatIII) BL cell lines was performed by polychromatic flow cytomery, based on expression pattern of CD19, CD10, CD38, CD27, and CD5 markers. Two cell subsets, CD19+CD10+ and CD19+CD10-, were defined in LatIII BL cell lines. In both subsets, the CD27 and CD5 cell surface expression was detected in a proportion of the cells.

Effect of Retinoic Acid on Fetus Reproductive Organ Mice (Mus musculus) Swiss Webster

Retinoic acid is like a steroid hormone that plays a role in embryo formation, proliferation of spermatogonia cells, ephitelial cells differentiation and organogenesis. Retinoic acid can influences seminiferous tubule formation during embryonic testis development and also play a role in the regulation of ovarian function and female reproductive tract by suppressing the hormones FSH receptor expression. The excessive use of retinoic acid caused abnormalities in the fetus. The result showed that there is the influence of retinoic acid on the developmet of mice fetal testes, for examples disruption of the formation of seminiferous tubules and tubules seemed to be hollow, spermatogonia cells are relatively few in number and caused Leydig cells count relatively more. While in the female fetus does not caused the formation of primordial follicles and disrupted the development of germinal ephitelial cells of fetal ovaries of female mice (mus musculus) Swiss Webster.

An Efficient Protocol for Cyclic Somatic Embryogenesis in Neem (Azadirachta indica A Juss.)

Neem is a highly heterozygous and commercially important perennial plant. Conventionally, it is propagated by seeds which loose viability within two weeks. Strictly cross pollinating nature of the plant causes serious barrier to the genetic improvement by conventional methods. Alternative methods of tree improvement such as somatic hybridization, mutagenesis and genetic transformation require an efficient in vitro plant regeneration system. In this regard, somatic embryogenesis particularly secondary somatic embryogenesis may offer an effective system for large scale plant propagation without affecting the clonal fidelity of the regenerants. It can be used for synthetic seed production, which further bolsters conservation of this tree species which is otherwise very difficult The present report describes the culture conditions necessary to induce and maintain repetitive somatic embryogenesis, for the first time, in neem. Out of various treatments tested, the somatic embryos were induced directly from immature zygotic embryos of neem on MS + TDZ (0.1 μM) + ABA (4 μM), in more than 76 % cultures. Direct secondary somatic embryogenesis occurred from primary somatic embryos on MS + IAA (5 μM) + GA3 (5 μM) in 12.5 % cultures. Embryogenic competence of the explant as well as of the primary embryos was maintained for a long period by repeated subcultures at frequent intervals. A maximum of 10 % of these somatic embryos were converted into plantlets.

Loss of P16/INK4A Protein Expression is a Common Abnormality in Hodgkin's Lymphoma

P16/INK4A is tumor suppressor protein that plays a critical role in cell cycle regulation. Loss of P16 protein expression has been implicated in pathogenesis of many cancers, including lymphoma. Therefore, we sought to investigate if loss of P16 protein expression is associated with lymphoma and/or any specific lymphoma subtypes (Hodgkin-s lymphoma (HL) and nonHodgkin-s lymphoma (NHL)). Fifty-five lymphoma cases consisted of 30 cases of HL and 25 cases of NHL, with an age range of 3 to 78 years, were examined for loss of P16 by immunohistochemical technique using a specific antibody reacting against P16. In total, P16 loss was seen in 33% of all lymphoma cases. P16 loss was identified in 47.7% of HL cases. In contrast, only 16% of NHL showed loss of P16. Loss of P16 was seen in 67% of HL patients with 50 years of age or older, whereas P16 loss was found in only 42% of HL patients with less than 50 years of age. P16 loss in HL is somewhat higher in male (55%) than in female (30%). In subtypes of HL, P16 loss was found exclusively in all cases of lymphocyte depletion, lymphocyte predominance and unclassified cases, whereas P16 loss was seen in 39% of mixed cellularity and 29% of nodular sclerosis cases. In low grade NHL patients, P16 loss was seen in approximately one-third of cases, whereas no or very rare of P16 loss was found in intermediate and high grade cases. P16 loss did not show any correlation with age or gender of NHL patients. In conclusion, the high rate of P16 loss seen in our study suggests that loss of P16 expression plays a critical role in the pathogenesis of lymphoma, particularly with HL.

Identification of Differentially Expressed Gene(DEG) in Atherosclerotic Lesion by Annealing Control Primer (ACP)-Based Genefishing™ PCR

Atherosclerosis was identified as a chronic inflammatory process resulting from interactions between plasma lipoproteins, cellular components (monocyte, macrophages, T lymphocytes, endothelial cells and smooth muscle cells) and the extracellular matrix of the arterial wall. Several types of genes were known to express during formation of atherosclerosis. This study is carried out to identify unknown differentially expressed gene (DEG) in atherogenesis. Rabbit’s aorta tissues were stained by H&E for histomorphology. GeneFishing™ PCR analysis was performed from total RNA extracted from the aorta tissues. The DNA fragment from DEG was cloned, sequenced and validated by Real-time PCR. Histomorphology showed intimal thickening in the aorta. DEG detected from ACP-41 was identified as cathepsin B gene and showed upregulation at week-8 and week-12 of atherogenesis. Therefore, ACP-based GeneFishing™ PCR facilitated identification of cathepsin B gene which was differentially expressed during development of atherosclerosis.

Cultural Integration as a Factor of Genesis of the Kazakh Nation in the Conditions of Multicultural Society

The article analyses historical aspects of the formation of the Kazakh nation in the conditions of the multicultural society. The authors underline cultural integration as a significant stage of the cultural advancement of the Kazakh nation. The transition to the modern-style houses, the adoption and development of the secular education gave a rise to the development of the society and culture on the whole.

Cultivation of Thymus by In Vitro And Hydroponics Combined Method

Our results showed that for the growth of qualitative seedling and vegetative raw material of ðó. marschallianus Willd. and T. serphyllum L. it is more profitable to use the in vitro and hydroponics combined method. In in vitro culture it is possible to do micro-propagation whole year with 98-99% rhizogenesis. 30000 micro-plants were obtained from one explant during 9 months. Hydroponic conditions provide the necessary microclimate for microplants where the survival rate without acclimatization was 93.3%. The essential oil content in hydroponic dry herb of both species in vegetative and blossom phase was 1.3% whereas in wild plants it was 1.2%, the content of extractive substances and vitamin C also exceeded wild plants. Our biochemical and radiochemical investigations indicated that the medicinal raw materials obtained from hydroponic and wild plants of Thymus species correspond to the demands of SPh XI, and the content of artificial radionuclides does not exceed the MACL.

Visfatin and Apelin Are New Interrelated Adipokines Playing Role in the Pathogenesis of Type 2 Diabetes Mellitus Associated Coronary Artery Disease in Postmenopausal Women

Visfatin and apelin are two new adipokines that recently gained a special interest in diabetes research. This study was conducted to study the interplay between these two adipokines and their correlation with other inflammatory and biochemical parameters in type 2 diabetic (T2D) postmenopausal women with CAD. Visfatin and apelin were measured by enzyme-linked immunoassay (ELISA). Visfatin was found to be significantly higher in the following groups: T2D patients without CAD, non-obese and obese T2D patients with CAD when compared to control group. Apelin was found to be significantly lower in non-obese and obese T2D patients with CAD when compared to control group. Visfatin and apelin were found to be significantly associated with each other and with other biochemical parameters. The current study provides evidence for the interplay between visfatin and apelin through the inflammatory milieu characteristic of T2D and their possible role in the pathogenesis of CAD complication of T2D. 

Proteins Length and their Phenotypic Potential

Mendelian Disease Genes represent a collection of single points of failure for the various systems they constitute. Such genes have been shown, on average, to encode longer proteins than 'non-disease' proteins. Existing models suggest that this results from the increased likeli-hood of longer genes undergoing mutations. Here, we show that in saturated mutagenesis experiments performed on model organisms, where the likelihood of each gene mutating is one, a similar relationship between length and the probability of a gene being lethal was observed. We thus suggest an extended model demonstrating that the likelihood of a mutated gene to produce a severe phenotype is length-dependent. Using the occurrence of conserved domains, we bring evidence that this dependency results from a correlation between protein length and the number of functions it performs. We propose that protein length thus serves as a proxy for protein cardinality in different networks required for the organism's survival and well-being. We use this example to argue that the collection of Mendelian Disease Genes can, and should, be used to study the rules governing systems vulnerability in living organisms.

Wasp Venom Peptides may play a role in the Pathogenesis of Acute Disseminated Encephalomyelitis in Humans: A Structural Similarity Analysis

Acute disseminated encephalomyelitis (ADEM) has been reported to develop after a hymenoptera sting, but its pathogenesis is not known in detail. Myelin basic protein (MBP)- specific T cells have been detected in the blood of patients with ADEM, and a proportion of these patients develop multiple sclerosis (MS). In an attempt to understand the mechanisms underlying ADEM, molecular mimicry between hymenoptera venom peptides and the human immunodominant MBP peptide was scrutinized, based on the sequence and structural similarities, whether it was the root of the disease. The results suggest that the three wasp venom peptides have low sequence homology with the human immunodominant MBP residues 85-99. Structural similarity analysis among the three venom peptides and the MS-related HLA-DR2b (DRA, DRB1*1501)-associated immunodominant MHC binding/TCR contact residues 88-93, VVHFFK showed that hyaluronidase residues 7-12, phospholipase A1 residues 98-103, and antigen 5 residues 109-114 showed a high degree of similarity 83.3%, 100%, and 83.3% respectively. In conclusion, some wasp venom peptides, particularly phospholipase A1, may potentially act as the molecular motifs of the human 3HLA-DR2b-associated immunodominant MBP88-93, and possibly present a mechanism for induction of wasp sting-associated ADEM.

Neurogenic Potential of Clitoria ternatea Aqueous Root Extract–A Basis for Enhancing Learning and Memory

The neurogenic potential of many herbal extracts used in Indian medicine is hitherto unknown. Extracts derived from Clitoria ternatea Linn have been used in Indian Ayurvedic system of medicine as an ingredient of “Medhya rasayana", consumed for improving memory and longevity in humans and also in treatment of various neurological disorders. Our earlier experimental studies with oral intubation of Clitoria ternatea aqueous root extract (CTR) had shown significant enhancement of learning and memory in postnatal and young adult Wistar rats. The present study was designed to elucidate the in vitro effects of 200ng/ml of CTR on proliferation, differentiation and growth of anterior subventricular zone neural stem cells (aSVZ NSC-s) derived from prenatal and postnatal rat pups. Results show significant increase in proliferation and growth of neurospheres and increase in the yield of differentiated neurons of aSVZ neural precursor cells (aSVZNPC-s) at 7 days in vitro when treated with 200ng/ml of CTR as compared to age matched control. Results indicate that CTR has growth promoting neurogenic effect on aSVZ neural stem cells and their survival similar to neurotrophic factors like Survivin, Neuregulin 1, FGF-2, BDNF possibly the basis for enhanced learning and memory.

Role of Oxidative DNA Damage in Pathogenesis of Diabetic Neuropathy

Oxidative stress is considered to be the cause for onset and the progression of type 2 diabetes mellitus (T2DM) and complications including neuropathy. It is a deleterious process that can be an important mediator of damage to cell structures: protein, lipids and DNA. Data suggest that in patients with diabetes and diabetic neuropathy DNA repair is impaired, which prevents effective removal of lesions. Objective: The aim of our study was to evaluate the association of the hOGG1 (326 Ser/Cys) and XRCC1 (194 Arg/Trp, 399 Arg/Gln) gene polymorphisms whose protein is involved in the BER pathway with DNA repair efficiency in patients with diabetes type 2 and diabetic neuropathy compared to the healthy subjects. Genotypes were determined by PCR-RFLP analysis in 385 subjects, including 117 with type 2 diabetes, 56 with diabetic neuropathy and 212 with normal glucose metabolism. The polymorphisms studied include codon 326 of hOGG1 and 194, 399 of XRCC1 in the base excision repair (BER) genes. Comet assay was carried out using peripheral blood lymphocytes from the patients and controls. This test enabled the evaluation of DNA damage in cells exposed to hydrogen peroxide alone and in the combination with the endonuclease III (Nth). The results of the analysis of polymorphism were statistically examination by calculating the odds ratio (OR) and their 95% confidence intervals (95% CI) using the ¤ç2-tests. Our data indicate that patients with diabetes mellitus type 2 (including those with neuropathy) had higher frequencies of the XRCC1 399Arg/Gln polymorphism in homozygote (GG) (OR: 1.85 [95% CI: 1.07-3.22], P=0.3) and also increased frequency of 399Gln (G) allele (OR: 1.38 [95% CI: 1.03-1.83], P=0.3). No relation to other polymorphisms with increased risk of diabetes or diabetic neuropathy. In T2DM patients complicated by neuropathy, there was less efficient repair of oxidative DNA damage induced by hydrogen peroxide in both the presence and absence of the Nth enzyme. The results of our study suggest that the XRCC1 399 Arg/Gln polymorphism is a significant risk factor of T2DM in Polish population. Obtained data suggest a decreased efficiency of DNA repair in cells from patients with diabetes and neuropathy may be associated with oxidative stress. Additionally, patients with neuropathy are characterized by even greater sensitivity to oxidative damage than patients with diabetes, which suggests participation of free radicals in the pathogenesis of neuropathy.

Community Detection-based Analysis of the Human Interactome Network

The study of proteomics reached unexpected levels of interest, as a direct consequence of its discovered influence over some complex biological phenomena, such as problematic diseases like cancer. This paper presents a new technique that allows for an accurate analysis of the human interactome network. It is basically a two-step analysis process that involves, at first, the detection of each protein-s absolute importance through the betweenness centrality computation. Then, the second step determines the functionallyrelated communities of proteins. For this purpose, we use a community detection technique that is based on the edge betweenness calculation. The new technique was thoroughly tested on real biological data and the results prove some interesting properties of those proteins that are involved in the carcinogenesis process. Apart from its experimental usefulness, the novel technique is also computationally effective in terms of execution times. Based on the analysis- results, some topological features of cancer mutated proteins are presented and a possible optimization solution for cancer drugs design is suggested.

Changes to Oxidative Stress Levels Following Exposure to Formaldehyde in Lymphocytes

Formaldehyde is the illegal chemical substance used for food preservation in fish and vegetable. It can promote carcinogenesis. Superoxide dismutases are the important antioxidative enzymes that catalyze the dismutation of superoxide anion into oxygen and hydrogen peroxide. The resultant level of oxidative stress in formaldehyde-treated lymphocytes was investigated. The formaldehyde concentrations of 0, 20, 40, 60, 80 and 120μmol/L were treated in human lymphocytes for 12 hours. After 12 treated hours, the superoxide dismutase activity change was measured in formaldehyde-treated lymphocytes. The results showed that the formaldehyde concentrations of 60, 80 and 120μmol/L significantly decreased superoxide dismutase activities in lymphocytes (P < 0.05). The change of superoxide dismutase activity in formaldehyde-treated lymphocytes may be the biomarker for detect cellular injury, such as damage to DNA, due to formaldehyde exposure.

Ultrasonic Evaluation of Bone Callus Growth in a Rabbit Tibial Distraction Model

Ultrasound is useful in demonstrating bone mineral density of regenerating osseous tissue as well as structural alterations. A proposed ultrasound method, which included ultrasonography and acoustic parameters measurement, was employed to evaluate its efficacy in monitoring the bone callus changes in a rabbit tibial distraction osteogenesis (DO) model. The findings demonstrated that ultrasonographic images depicted characteristic changes of the bone callus, typical of histology findings, during the distraction phase. Follow-up acoustic parameters measurement of the bone callus, including speed of sound, reflection and attenuation, showed significant linear changes over time during the distraction phase. The acoustic parameters obtained during the distraction phase also showed moderate to strong correlation with consolidated bone callus density and micro-architecture measured by micro-computed tomography at the end of the consolidation phase. The results support the preferred use of ultrasound imaging in the early monitoring of bone callus changes during DO treatment.

Function of miR-125b in Zebrafish Neurogenesis

MicroRNAs are an important class of gene expression regulators that are involved in many biological processes including embryogenesis. miR-125b is a conserved microRNA that is enriched in the nervous system. We have previously reported the function of miR-125b in neuronal differentiation of human cell lines. We also discovered the function of miR-125b in regulating p53 in human and zebrafish. Here we further characterize the brain defects in zebrafish embryos injected with morpholinos against miR-125b. Our data confirm the essential role of miR-125b in brain morphogenesis particularly in maintaining the balance between proliferation, cell death and differentiation. We identified lunatic fringe (lfng) as an additional target of miR-125b in human and zebrafish and suggest that lfng may mediate the function of miR-125b in neurogenesis. Together, this report reveals new insights into the function of miR- 125b during neural development of zebrafish.