A comparative analysis of methylation status of tumor suppressor genes in paired biopsy and serum samples from cervical cancer patients among north indian population

Tumor-specific genetic or epigenetic alterations have been detected in serum DNA in case of various types of cancers. In breast cancer, the detection of tumor suppressor gene hypermethylation has been reported in several body fluids. Promoter hypermethylation of some genes like MYOD1, CALCA, hTERT, etc. has also been detected in serum samples from cervical cancer. The present study is the first report on the comparison of promoter hypermethylation of tumor suppressor genes like p14, p15, p16, p21, p27, p57, p53, p73, RARβ2, FHIT, DAPK, STAT1, and RB1 genes in paired biopsy and serum samples from cervical cancer patients among north Indian population. This is also the first report on the hypermethylation of these genes in serum samples from cervical cancer patients among north Indian population. According to the results of the present study, promoter hypermethylation of these genes can also be detected in serum samples of cervical cancer patients. The sensitivity of detection of promoter hypermethylalion in serum samples of cervical cancer patients as compared to paired biopsy samples was found to be around 83.3%. It was observed that promoter hypermethylation was mainly observed in the serum samples in the higher stages and very rarely in the lower stages. The present study clearly showed that serum of patients with cervical cancer can also be used to study methylated genes as biomarkers.     

Oxidase-Peroxidase Method of Ethanol Assay in Fermented Musts and Wine Products

A new alcohol oxidase-peroxidase method of ethanol assay in fermented musts and wine products is described and compared to conventional methods routinely used in winemaking. The sensitivity, accuracy, and reliability of this method were determined. The results of ethanol determination in fermented musts and wines correlated well with the data obtained by refractometry (correlation coefficient R = 0.9595, p < 0.0001) and densitometry (correlation coefficient R = 0.9384, p < 0.0001). The proposed method is less time- and labor-consuming and allows simultaneous analysis of a series of wine samples.     

Contents of Heavy Metals in Chinese Edible Herbs: Evidence from a Case Study of Epimedii Folium

Toxic heavy metal contamination in Chinese edible herbs has raised a worldwide concern. In this study, heavy metals in Epimedii Folium, an edible medicinal plant in China, were quantitatively analyzed. Variations of heavy metals in different species, in various organs (i.e., leaves, stems, and roots), in wild-growing and cultivated plants, and in 35 market samples of Epimedii Folium, were systematically investigated. In all of Epimedium samples, Hg (mercury) was not detectable (0.00 μg/g). Four species, Epimedium pubescens, Epimedium sagittatum, Epimedium brevicornu, and Epimedium wushanense, were found to contain Cu (copper) and Pb (lead). And contents of Cu and Pb in E. brevicornu were significantly higher than those in other species (P < 0.01). In wild-growing and cultivated Epimedium plants, Cd (cadmium) and As (arsenic) were not detectable, and concentrations of Cu and Pb in wild-growing plants were significantly higher than those in cultivated plants (P < 0.01). Cd was not detectable in leaves, roots, and stems, while organ specificity was apparent in the distribution of Cu, As, and Pb. And the highest levels of Cu and Pb were observed in roots and leaves, respectively. In Chinese markets, several samples of Epimedii Folium contained excessive Cu, Cd, As, and Pb beyond the national permissible limits. In summary, there was a large variation of heavy metals among Epimedii Folium samples, and Cu and Pb were the most important heavy metals contaminating the edible medicinal plant. Application of Epimedii Folium to drug and food industries will need to focus more on toxic heavy metal contamination.     

Effect of fibrinogen on platelet reactivity measured by the VerifyNow P2Y<Subscript>12</Subscript> assay

The VerifyNow assay is based upon the ability of activated platelets to cross-link beads coated with fibrinogen. However, fibrinogen is an abundant protein of blood, and therefore it may affect test results by competing with fibrinogen of beads for binding to platelets. To test this assumption, we assessed the influence of artificial alteration of fibrinogen level in blood samples obtained from donors (n = 9) and patients on clopidogrel therapy (n = 8) on the results of the VerifyNow P2Y₁₂ assay. Fibrinogen level was altered by adding to blood samples 1/10 volume of fibrinogen solution (10.56 g/liter) or corresponding buffer. Relative to baseline, addition of buffer significantly increased platelet reactivity, whereas addition of fibrinogen decreased it. Analysis of the relationship between change in platelet reactivity values (dBase and dPRU) and change in fibrinogen concentration (dFg) revealed strong negative correlations: dBase =–63.3 × dFg–27.1 (r =–0.924, p < 0.0005) and dPRU =–54.4 × dFg–21.8 (r =–0.764, p < 0.0005). Thus, the results of our experiments suggest that: (i) blood fibrinogen strongly influences results of the VerifyNow P2Y₁₂ assay, and (ii) correcting for fibrinogen effect may be needed to improve the accuracy of the test in the measuring of antiplatelet effect of clopidogrel therapy.     

Mammaglobin in peripheral blood and the tumor of breast cancer patients

Currently, no molecular biological markers do exist for early diagnosis of breast cancer. One of the possible candidates for the marker of early breast cancer is mammaglobin (MGB1) or SCGB2A2 (secretoglobin, family 2A, member 2), characterized by the maximal expression level in early breast cancer. Using the RT-PCR method MGB1 mRNA expression was examined in 57 tumor tissue samples and 57 samples of morphologically non-malignant tissue (MNT) of breast cancer (BC) patients. Specificity and sensitivity of the MGB1 mRNA assay in peripheral blood of BC patients was evaluated by nested PCR. 169 blood samples (from 95 BC patients, 22 from patients with benign breast tumors, 28 from patients with tumors of other localizations, and 24 samples from healthy donors) have been analyzed. MGB1 expression was significantly higher in BC tissue samples compared to MNT (p = 0.0019). The maximal expression level was in the samples T1 (p = 0.013), stage I BC (p = 0.037), GI (p = 0.0019). MGB1 expression positively correlated with expression of estrogen (p = 0.034) and progesterone (p = 0.0004) receptors. Sensitivity and specificity of the MGB1 mRNA assay in peripheral blood were 60.6 and 92.3%, respectively. Expression of MGB1 was higher in BC than MNT and it decreased during BC progression. The sensitivity and specificity of the MGB1 mRNA assay may be used as an additional diagnostic method.     

<Emphasis Type="Italic">Saccharothrix ghardaiensis</Emphasis> sp. nov., an actinobacterium isolated from Saharan soil

The taxonomic position of a new Saccharothrix strain, designated MB46^T, isolated from a Saharan soil sample collected in Mzab region (Ghardaïa province, South Algeria) was established following a polyphasic approach. The novel microorganism has morphological and chemical characteristics typical of the members of the genus Saccharothrix and formed a phyletic line at the periphery of the Saccharothrix espanaensis subcluster in the 16S rRNA gene dendrograms. Results of the 16S rRNA gene sequence comparisons revealed that strain MB46^T shares high degrees of similarity with S. espanaensis DSM 44229^T (99.2%), Saccharothrix variisporea DSM 43911^T (98.7%) and Saccharothrix texasensis NRRL B-16134^T (98.6%). However, the new strain exhibited only 12.5–17.5% DNA relatedness to the neighbouring Saccharothrix spp. On the basis of phenotypic characteristics, 16S rRNA gene sequence comparisons and DNA-DNA hybridizations, strain MB46^T is concluded to represent a novel species of the genus Saccharothrix, for which the name Saccharothrix ghardaiensis sp. nov. (type strain MB46^T = DSM 46886^T = CECT 9046^T) is proposed.     

Intrapopulation Polymorphism of the Pine Marten (<Emphasis Type="Italic">Martes martes</Emphasis>, Carnivora,Mustelidae) from Tver Oblast

The intrapopulation polymorphism of the pine marten (Martes martes L. 1758) was assessed on the basis of studying five samplings of skulls, three of which were collected on the same territory at different times. The geographical distance between the three sampling areas varied from 60 to 240 km. Study of the material using the phenetic (250 skulls), morphometric (151 skulls), and molecular genetic (21 skin samples) approaches allowed us to characterize the pine marten as a species with a low level of polymorphism. The genetic diversity (π = 0.0069 ± 0.0041 and H = 0.90 ± 0.039) is determined by ancient demographic processes that took place in the Pleistocene and Holocene and are related to the ice-age history of this species, its expansion, and the formation of the population on the territory under study. Low levels of phenetic (μ = 1.48 ± 0.056, h = 0.14 ± 0.033) and morphological (CV = 3.37 ± 0.41) diversity as well as a low level of sexual dimorphism (Isd = 9.4) indicate that the pine marten occupies a specific and relatively narrow ecological niche and is involved in weak competition with smaller mustelid species.     

Antioxidative effect of CLA diet and endurance training in liver and skeletal muscles of rat

The purpose of this study is to investigate the effects of conjugated linoleic acid (CLA) supplementation and endurance exercise on the oxidative/anti-oxidative status in rat liver and skeletal muscles. Sprague-Dawley male rats were randomly divided into HS (high-fat diet sedentary group, n = 8), CS group (CLA supplemented sedentary group, n = 8), and CE group (CLA supplemented exercise group, n = 8). For CLA supplementation, 1.0% CLA was substituted for dietary fat. For endurance exercise, the rats swam for 60 min a day, 5 days a week for 8 weeks. The MDA content, Cu, Zn-SOD and Mn-SOD expression in the soleus muscle (SOM) of the CE group improved significantly compared to the HS (p < 0.01) and CS groups (p < 0.05). Moreover, Mn-SOD expression in both the SOM and extensor digitorum longus muscle (EDL) of the CS were enhanced significantly compared to the HS (p < 0.05). From these results, it was suggested that CLA supplementation under the endurance exercise condition may improve the oxidative status by decreasing the MDA content via potential scavenging of Cu,Zn-SOD, and Mn-SOD protein in red muscle, respectively. Therefore, our study demonstrated long-term endurance exercise with CLA supplementation plays a crucial role for maintenance of antioxidative properties in the skeletal muscle of rat.     

Molecular profiling of fungal assemblages in the healthy and infected roots of <Emphasis Type="Italic">Decalepis arayalpathra</Emphasis> (J. Joseph &amp; V. Chandras) Venter,an endemic and endangered ethnomedicinal plant from Western Ghats,India

Decalepis arayalpathra, an endangered, endemic ethnomedicinal plant from southern Western Ghats, India, is targeted for its aromatic and medicinal properties. This study aimed at to identify fungal endophyte populations associated with healthy and diseased roots of this perennial shrub. Healthy and rotted root samples of D. arayalpathra were collected, fungal endophytes assemblages were identified both by culture-dependent and culture-independent approaches, further sequenced and the retrieved sequences were analysed with the reference sequences in GenBank to know their phylogenetic relationships. Analysis of the ITS rDNA region generated 24 different Ascomycota and three Basidiomycota taxa. Trichoderma sp. was most abundant in healthy and diseased root samples, while Penicillium and Aspergillus were confined to healthy roots. Furthermore, Fusarium solani, Fusarium oxysporum and Mucor velutinosus were found to be the most frequent fungi identified from the rotted root samples, thus substantiated to be the cause for D. arayalpathra decline in the wild. Interestingly, the strains assigned to Fusarium sp. were isolated from diseased roots showing typical clearly visible symptoms, such as a severe brown discolouration on the taproot. Molecular profiling of all the pure fungal isolates, viz., Trichoderma, Penicillium, Aspergillus, Fusarium and Mucor, revealed high sequence similarities (≥ 98 %) to corresponding reference sequences. Sequencing of Trichoderma pure cultures isolated from healthy and diseased roots revealed sequence similarities to Trichoderma harzianum, T. hamatum, T. koningiopsis, T. asperellum, T. pubescens and Hypocrea sp. This confirms the morphological examinations, as Hypocrea is the teleomorph stage of Trichoderma sp. This study signifies the first work pertaining to the taxonomy of the fungal endophytic community of D. arayalpathra, and the results reported in this work may help to ascertain the cause of root rot disease often perceived in D. arayalpathra. Also, it could be useful to identify the promising endophytic communities against the root rot diseases occurring in D. arayalpathra.     

Association analysis of alcohol metabolizing enzymes <Emphasis Type="Italic">ADH1B,ADH7, CYP2E1</Emphasis> gene polymorphism with risk for coronary atherosclerosis

The allele and genotype distribution of two alcohol dehydrogenase genes ADH1B (exon 3 polymorphism A/G (47His)), ADH7 (intron 5 polymorphism G/C) and cytochrome P450 2E1 gene (CYP2E1; 5′-flanking region G/C and intron 6 T/A polymorphisms) were examined in Russian (Tomsk, n = 125) healthy population and in coronary atherosclerosis patients (CA, n = 92). The genotype frequencies followed the Hardy-Weinberg equilibrium and the alleles were in linkage equilibrium or gametic equilibrium in the control sample. Only two CYP2E1 gene polymorphisms were in linkage disequilibrium. The frequencies of the derived alleles at ADH1B ^* G (+MslI) allele, CYP2E1 ^* C2 (+PstI) allele and CYP2E1 ^* C (-DraI) allele were 8.48 ± 1.86, 1.20 ± 0.69, and 10.00 ± 1.90%, respectively. The ADH7 gene polymorphism showed a high level of heterozygosity; the frequency of the ADH7 ^* C (-StyI) allele was 44.58 ± 3.21%. A significantly higher frequency of CYP2E1 PstI C2 allele has been revealed in the CA group (P = 0.043; OR = 4.23; 95% CI 1.03–20.01). The tendency to significant effect of A1A2 genotype in ADH1B MslI polymorphism was observed for systolic blood pressure in the control group (P = 0.068). The statistically significant two-way interaction effects of ADH7 StyI and CYP2E1 DraI on diastolic blood pressure (P = 0.029) and on the serum high density lipoprotein level (P = 0.042) were also revealed. Association of A1A2 genotype in ADH1B MslI polymorphism with reduced amount in a serum of a very low density lipoprotein level (P = 0.045) have also been shown. This may result from multifunctional activity of alcohol metabolizing enzymes and their involvement in many metabolic and free radical reactions in the body.     

Genetic differentiation in the polyploid complex of <Emphasis Type="Italic">Suaeda corniculata</Emphasis> (C.A. Mey.) Bunge in Eastern Siberia

Data on differentiation of the polyploid complex of Suaeda corniculata (C.A. Mey.) Bunge in East Siberia based on variability of morphological characteristics, karyological analysis, and genetic polymorphism in inter simple sequence repeat (ISSR) markers were given for the first time. It was established that the samples studied belonged to three cytotypes: diploids (2n = 2x = 18), tetraploids (2n = 4x = 36) and hexaploids (2n = 6x = 54). The analysis of variability of morphological characteristics by the method of principal coordinates showed division of the samples into three separate groups. On the basis of morphological and molecular-genetic methods, a genetically differentiated diploid population was revealed in Yakutia.     

Phenotypic and genotypic typing of<Emphasis Type="Italic">Salmonella enterica</Emphasis> serovar Enteritidis isolates from poultry farms environments in Tunisia

Salmonella enterica serovar Enteritidis is one of the majorSalmonella serovars which may cause animal infections and human salmonellosis. In this study, two hundred forty five samples (faeces, water and environmental swabs) were taken from eight poultry farms localized in different geographical areas of Tunisia. We foundSalmonella serovar Enteritidis (16 strains),Salmonella typhimirium (2),Salmonella scharzengrund (2), andSalmonella braenderup (1).Salmonella Enteritidis strains were characterized by pulsed field gel electrophoresis (PFGE) analysis, plasmid analysis and antibiotic resistance profiles.XbaI PFGE analysis revealed two PFGE types and plasmid profiling identified four plasmid types. The majority of isolates were susceptible to all antibiotic tested. The combined use of phenotypic and genotypic methods indicates the spread of a particularSalmonella Enteritidis clone. This clone is highly related to a major world-wide clone identified in many other countries.     

On the minimum electron transport coefficients in tokamaks in the range of low collision frequencies

There are two close empirical scalings, namely, the T-11 and neo-Alcator ones, that provide correct estimates for the energy confinement time in tokamaks in ohmic heating regimes in the linear part of the dependence τ _E (\(\bar n_e \)) in the range of low values of \(\bar n_e \) and 〈ν _e ^* 〉 ≤ 1. The similar character of electron energy confinement in this range, which expands with increasing magnetic field B ₀, has stimulated the search for dimensionless parameters and simple physical models that would explain the experimentally observed dependences χ _e ～ 1/n _e and τ _Ee ～ \(\bar n_e \). In 1987, T. Okhawa showed that the experimental data were satisfactorily described by the formula χ_e⊥ = (c ²/ω _pe ² )ν _e /qR, in deriving of which the random spatial leap along the radius r on the electron trajectory was assumed to be the same as that in the coefficient of the poloidal field diffusion, while the repetition rate of these leaps was assumed to be ν _e /qR. In 2004, J. Callen took into account the decrease in the fraction of transient electrons with increasing toroidal ratio ? = r/R and corrected the coefficient c ²/ω _pe ² in Okhawa equation by the factor σ _‖ ^Sp /σ _‖ ^neo . If one takes into account this correction and assumes that the frequency of the stochastic process is equal to the reciprocal of the half-period of rotation of a trapped electron along its banana trajectory, then the resulting expression for χ_e⊥ will coincide with the T-11 scaling: χ _e ^an ∞ ?^1.75(T _e /A _i )^0.5/(n _e qR) at A i = 1. If the same stochastic process also involves ions, it may result in the opening of the orbit of a trapped ion at the distance ～(c/ω _pe )(m _i /m _e )^1/4. In this case, the calculated coefficient of electron and ion diffusion D is close to D ^an ≈ χ _e ^an /2.     

Chaotic Dynamics of Neuromuscular System Parameters and the Problems of the Evolution of Complexity

The evolution rate v(t) varies among diverse biosystems, but a general theory can be formulated when the dynamics of the biosystem stater x = x(t) = (x₁, x₂, x _m ) ^T is considered in the m-dimensional space of states. A mathematical approach is proposed for evaluating such processes and describes the processes in terms of particular chaos of the statistical distribution functions f(x). In the case of complex multicomponent systems with a high dimension number m (m ?1) of the phase space of states, we propose using pairwise comparison matrices of samples x(t) when homeostasis is constant and calculating the parameters of quasiattractors. The Glensdorff–Prigogine thermodynamic approach to estimating evolution is inefficient in assessing the third-type systems, while it is applicable and the Prigogine theorem works at the level of molecular systems. Alterations in the state of the human neuromuscular system were found to lead to chaotic changes in the statistical functions f(x) in tremor recording samples, while quasiattractor parameters demonstrate a certain regularity.     

Association between serum somatostatin levels and glucose-lipid metabolism in the Jino ethnic minority and Han Chinese population

We aim to investigate the relationship between serum somatostatin(SST) levels and glucose-lipid metabolism at various stages of glucose tolerance in the Jino ethnic minority(n=111) and Han population(n=113) of Yunnan Province, southwest China.Anthropometric parameters and biochemical traits were measured. Serum SST and plasma glucagon levels were tested. Participants were divided into three subgroups: isolated fasting hyperglycemia(IFH), isolated post challenge hyperglycemia(IPH)and normal glucose tolerance(NGT). SST levels were found lower while glucagon levels were significantly higher in the Jino ethnic with IPH(P=0.0026 and P=0.0069, respectively). Fasting glucose and high density lipoprotein-cholesterol(HDL-C)levels were higher(P=0.0055 and P=0.0021, respectively) and fasting insulin levels and homeostasis model assessments β-cell function were lower(P=0.0479 and P=0.0007, respectively) in the Jino population. After adjusting for confounding factors, the serum SST level was associated with glucagon(P0.0001) in both populations. The SST level was correlated with fasting Cpeptide(P=0.0267) in Jino and HDL-C levels in Han(P=0.0079). Our findings suggest that serum SST levels and plasma glucagon levels may vary in subjects with IPH between two ethnics.     

Chromosomes of Ichneumonidae (Hymenoptera): New results and conclusions

New data were obtained on the karyotypes of 12 parasitic wasp species of the family Ichneumonidae: Scambus detritus (2n = 28), S. vesicarius (2n = 28), Pimpla contemplator (2n = 36), Theronia atalantae (2n = 24), Stenomacrus sp. 2 (n = 18, 2n = 36), Phygadeuon sp. 3 (2n = 32), Ichneumon memorator (2n = 24), I. vorax (2n = 22), Lymantrichneumon disparis (2n = 26), Orthopelma mediator (n = 15, 2n = 30), Lissonota buccator (2n = 22), and Trichionotus flexorius (2n = 16). Chromosomes of Anomaloninae were studied for the first time. The data on the karyotype structure of the subfamily Orthopelmatinae were revised. Various aspects of chromosome variability in Ichneumonidae are discussed.     

Decolorization of textile dye RB19 using volcanic rock matrix immobilized <Emphasis Type="Italic">Bacillus thuringiensis</Emphasis> cells with surface displayed laccase

A triplicate volcanic rock matrix–Bacillus thuringiensis–laccase WlacD (VRMs–Bt–WlacD) dye decolorization system was developed. WlacD was displayed on the B. thuringiensis MB174 cell surface to prepare a whole-cell laccase biocatalyst by using two repeat N-terminal domains of autolysin Mbg (Mbgn)₂ as the anchoring motif. Immunofluorescence microscopic assays confirmed that the fusion protein (Mbgn)₂–WlacD was anchored on the surface of the recombinant B. thuringiensis MB174. After optimization by a single factor test, L ₉(3⁴)-orthogonal test, Plackett–Burman test, steepest ascent method, and Box–Behnken response surface methodology, the whole-cell specific laccase activity of B. thuringiensis MB174 was improved to 555.2 U L^?1, which was 2.25 times than that of the primary culture condition. Optimized B. thuringiensis MB174 cells were further adsorbed by VRMs to prepare VRMs–Bt–WlacD, an immobilized whole-cell laccase biocatalyst. Decolorization capacity of as-prepared VRMs–Bt–WlacD toward an initial concentration of 500 mg L^?1 of an textile dye reactive blue 19 (RB19) aqueous solution reached 72.36% at a solid-to-liquid ratio of 10 g–100 mL. Repeated decolorization-activation operations showed the high decolorization capacity of VRMs–Bt–WlacD and have the potential for large-scale or continuous operations.     

The spectrum of mutations in genes associated with resistance to rifampicin,isoniazid, and fluoroquinolones in the clinical strains of <Emphasis Type="Italic">M. tuberculosis</Emphasis> reflects the transmissibility of mutant clones

To study the transmissibility of drug resistant mutant clones, M. tuberculosis samples were isolated from the patients of the clinical department and the polyclinic of the Central TB Research Institute (n = 1455) for 2011–2014. A number of clones were phenotypically resistant to rifampicin (n = 829), isoniazid (n = 968), and fluoroquinolones (n = 220). We have detected 21 resistance-associated variants in eight codons of rpoB, six variants in three codons of katG, three variants in two positions of inhA, four variants in four positions of ahpC, and nine variants in five codons of gyrA, which were represented in the analyzed samples with varied frequencies. Most common mutations were rpoB 531 Ser→Leu (77.93%), katG 315 (Ser→Thr) (94.11%), and gyrA 94 (Asp→Gly) (45.45%). We found that the mutations at position 15 of inhA (C→T) (frequency of 25.72%) are commonly associated with katG 315 (Ser→Thr). This association of two DNA variants may arise due to the double selection by coexposure of M. tuberculosis to isoniazid and ethionamide. The high transmissibility of mutated strains was observed, which may be explained by the minimal influence of the resistance determinants on strain viability. The high transmissibility of resistant variants may also explain the large populational prevalence of drug-resistant TB strains.