Minor Region Research Articles

Two DNA binding modes of a zinc-metronidazole and biological evaluation as a potent anti-cancer agent

A complex of metronidazole (MTZ) with zinc ion was synthesized and characterized by UV-Vis, Fourier transform infrared (FT-IR), 1H-NMR, X-ray crystallography and thermal gravimetric-differential thermal analysis (TG-DTA). The cytotoxicity effect of the synthesized complex investigated over SKNMC, A549, MCF-7, and MCDK cell lines and the results have shown that it has high cytotoxic potential over cancer cell lines. In order to clarify the mechanism of cell cytotoxicity, the oxidative stress and binding of the complex to the calf thymus-DNA studied by evaluating the intrinsic binding constant and defining thermodynamic parameters of complex over the DNA accompanying with in silico molecular modeling method. For this purpose, the complex optimized at the B3LYP/LANL2DZ level and docked over the DNA structure. The results revealed that the metronidazole-zinc complex interacted with DNA via hydrogen binding and electrostatic interaction to the minor groove region and phosphate backbone of DNA, respectively.

Structural, rheological and antioxidant properties of pectins from Equisetum arvense L. and Equisetum sylvaticum L.

The pectins were isolated from sterile stems of E. arvense (EA, yield 5.9%) and E. sylvaticum (ES, yield 4.8%) (Equisetaceae) using ammonium oxalate extraction after preliminary treatment with dilute HCl (рH 4.0). The pectins possessed high molecular weight (Mw, 340–360 kDa), high GalA content (ca. 85%), low degrees of methyl-esterification (14–16%) and acetylation (3–8%). NMR analysis indicated extensive regions of partially methyl-etherified and 3-O-acetylated HG and minor regions of low branched RG in the fragment isolated after hydrolysis of pectin EA by pectinase. Pectin EA produced a higher viscosity solution, formed a stronger and more rigid ionotropic hydrogel than pectin ES. The pectins scavenged DPPH and hydroxyl radicals, but not the superoxide radical and hydrogen peroxide. Phenolic compounds (0.11 and 0.23%) associated with polysaccharide moieties were apparently responsible for the differences in the anti-DPPH scavenging activity of pectins EA and ES (63 and 49%). The findings suggested that pectin from E. arvense should be more perspective than pectin from E. sylvaticum on their use as components of wound healing remedies.

Structural studies of biologically active pectin-containing polysaccharides of pomegranate Punica granatum

The polysaccharide PGW was isolated from peels and membranes of fruits of pomegranate P. granatum by hot water extraction. The methods of ion exchange chromatography, partial acid hydrolysis, enzymatic hydrolysis and NMR spectroscopy were employed to establish the major elements of its structure. It was shown that this polysaccharide contained polymers with different structures, the main components among which were pectic polysaccharides, represented mainly by highly methyl-esterified and lowly acetylated 1,4-α-d-galactopyranosyluronan and a minor amount of partially 2-O- and/or 3-O-acetylated RG-I. The side carbohydrate chains of the branched region of RG-I were mainly represented by terminal, 5-O-, 3-O-, 2,5-di-O-, 3,5-di-O- and 2,3,5-tri-O-substituted α-l-Araf residues indicating the presence of branched 1,5-α-l-arabinan and minor regions of 1,4-β-d-galactan or arabinogalactan type I. The degree of methylation of the isolated pectins varied depending on the method of treatment. As a result, peels and membranes of pomegranate fruits can be recommended as a source of highly and lowly methyl-esterified pectic polysaccharides.

Size distribution of trilayer graphene flakes obtained by electrochemical exfoliation of graphite: Effect of the synthesis parameters

Flake size distribution, one of the important aspects to be addressed for large-scale application of graphene, is largely ignored in the research on graphene synthesis. For the liquid phase methods of graphene synthesis, a systematic study is complicated by a scatter in the number of layers of the obtained graphene. We use, a recently reported, electrochemical exfoliation method for obtaining completely trilayer graphene suspension to study the change in trilayer flake size distribution with a change in the synthesis conditions. XRD analysis shows that the exfoliation current density affects the domain size of the intermediate graphite bisulfate (GB) even though the DLS-measured size of the exfoliated GB particles were similar. The difference in the GB domain size resulted in a very different average size as well as the tail of the size distribution of the final graphene flakes obtained by the ultrasonic cleaving of the intermediate GB particles. The power density employed in the ultrasonic cleaving step and the speed of the centrifugation employed to remove impurities from the graphene suspension also affect the flake size distribution. A trilayer graphene suspension of average flake area ∼15 μm2 with the flake size distribution slightly skewed to the right (larger size) and tail like a normal distribution is obtained by optimizing these three synthesis variables. The graphene flakes also contained minor four-layer regions and the effect of the three synthesis variables on their distribution is also studied.

Phenotypic and genotypic analyses of an attenuated porcine reproductive and respiratory syndrome virus strain after serial passages in cultured porcine alveolar macrophages.

The porcine reproductive and respiratory syndrome virus (PRRSV) is a globally ubiquitous swine viral pathogen that causes major economic losses worldwide. We previously reported an over-attenuated phenotype of cell-adapted PRRSV strain CA-2-P100 in vivo. In the present study, CA-2-P100 was serially propagated in cultured porcine alveolar macrophage (PAM) cells for up to 20 passages to obtain the derivative strain CA-2-MP120. Animal inoculation studies revealed that both CA-2-P100 and CA-2-MP120 had decreased virulence, eliciting weight gains, body temperatures, and histopathologic lesions similar to those in the negative control group. However, compared to CA-2-P100 infection, CA-2-MP120 yielded consistently higher viremia kinetics and enhanced antibody responses in pigs. All pigs inoculated with CA-2-MP120 developed viremia and seroconverted to PRRSV. During 20 passages in PAM cells, CA-2-MP120 acquired 15 amino acid changes that were mostly distributed in nsp2 and minor structural protein-coding regions. Among these changes, 6 mutations represented reversions to the sequences of the reference CA-2 and parental CA-2-P20 strains. These genetic drifts may be hypothetical molecular markers associated with PRRSV macrophage tropism and virulence. Our results indicate that the PAM-passaged CA-2-MP120 strain is a potential candidate for developing a live, attenuated PRRSV vaccine.

Comprehensive Assessment of Genotype Imputation Performance

Genotype imputation is a process of estimating missing ge­notypes from the haplotype or genotype reference panel. It can effectively boost the power of detecting single nucleotide polymorphisms (SNPs) in genome-wide association studies, integrate multi-studies for meta-analysis, and be applied in fine-mapping studies. The performance of genotype imputation is affected by many factors, including software, reference selection, sample size, and SNP density/sequencing coverage. A systematical evaluation of the imputation performance of current popular software will benefit future studies. Here, we evaluate imputation performances of Beagle4.1, IMPUTE2, MACH+Minimac3, and SHAPEIT2+ IM­PUTE2 using test samples of East Asian ancestry and references of the 1000 Genomes Project. The result indicated the accuracy of IMPUTE2 (99.18%) is slightly higher than that of the others (Beagle4.1: 98.94%, MACH+Minimac3: 98.51%, and SHAPEIT2+IMPUTE2: 99.08%). To achieve good and stable imputation quality, the minimum requirement of SNP density needs to be > 200/Mb. The imputation accuracies of IMPUTE2 and Beagle4.1 were under the minor influence of the study sample size. The contribution extent of reference to genotype imputation performance relied on software selection. We assessed the imputation performance on SNPs generated by next-generation whole genome sequencing and found that SNP sets detected by sequencing with 15× depth could be mostly got by imputing from the haplotype reference panel of the 1000 Genomes Project based on SNP data detected by sequencing with 4× depth. All of the imputation software had a weaker performance in low minor allele frequency SNP regions because of the bias of reference or software. In the future, more comprehensive reference panels or new algorithm developments may rise up to this challenge.

Observation of distorted Maxwell-Boltzmann distribution of epithermal ions in LHD

A distorted Maxwell-Boltzmann distribution of epithermal ions is observed associated with the collapse of energetic ions triggered by the tongue shaped deformation. The tongue shaped deformation is characterized by the plasma displacement localized in the toroidal, poloidal, and radial directions at the non-rational magnetic flux surface in toroidal plasma. Moment analysis of the ion velocity distribution measured with charge exchange spectroscopy is studied in order to investigate the impact of tongue event on ion distribution. A clear non-zero skewness (3rd moment) and kurtosis (4th moment –3) of ion velocity distribution in the epithermal region (within three times of thermal velocity) is observed after the tongue event. This observation indicates the clear evidence of the distortion of ion velocity distribution from Maxwell-Boltzmann distribution. This distortion from Maxwell-Boltzmann distribution is observed in one-third of plasma minor radius region near the plasma edge and disappears in the ion-ion collision time scale.

MOLECULAR CHARACTERIZATION OF INFECTIOUS BURSAL DISEASE VIRUS FROM COMMERCIAL POULTRY IN PAKISTAN

Infectious bursal disease (IBD) is an immunosuppressive disease of young, growing chickens which results in impaired growth or mortality of rearing flocks. In the current era there is a re-emergence of very virulent Infectious Bursal Disease Viruses (vvIBDV) and classical variant (cv) IBDV strains which increased the financial losses of poultry industry worldwide. Recent studies were conducted to characterize the existing vvIBDVs prevailing in Pakistan. The suspected samples were collected from the field outbreaks during the period from 2014–2017. IBDV was detected by RT-PCR. The sequences of VP2 gene (hyper variable region) were determined and available details were aligned with sequences submitted inGenBank. Phylogenetic analysis reveals that both vvIBDV and classical variant strains were circulating in different regions of Pakistan. In Indo-Pak isolates, the presence of virulent markers, amino acids (A222, I242, Q253, I256 and S299) and “Serine rich- heptapeptide” indicated the presence of very virulent viruses. The presence of T284A isan indicator of vvIBDVs in local poultry farms. More than 99% similarity of Pakistani isolates with Indian sequences reflects the trans-boundary spread of IBD. In recent studies amino acid, Glutamine (Q) is present at position 221 (as reported in previous studies) rather than Histidine (H) in Pakistani sequences. It is investigated that Glutamic acid (E) is located at position 300 in minor hydrophilic region III of VP2 protein in all reported Pakistani isolates. It is the unique feature of indigenous strains. This study will be useful in understanding the origin and pathotypes of IBDV circulating in Pakistan.

Application of Real-time Quantitative PCR in Detecting Atypical BCR/ABL mRNA Transcripts in Chronic Myelocytic Leukemia

To detect atypical BCR/ABL mRNA transcript by real-time quantitative PCR in CML patients without e13a2/e14a2,e19a2 or e1a2 transcripts, and investigate its value of clinical application. Twelve cases of CML with positive for t(9;22) translocation, but negative for common major and minor breakpoint cluster regions comfirmed by chromosome karyotyping or FISH analysis, were collected from July 2012 to December 2015. These 12 cases were then detected for b2a3(e13a3), b3a3(e14a3), e6a2, e8a2 and e1a3 fusion variants by real-time quantitative PCR. Among 12 cases 4 variant transcripts were detected, including e1a3 in 1 case (8.33%), e8a2 in 2 cases (16.67%), b2a3 in 5 cases (41.67%) and b3a3 in 4 cases (33.33%), with total positivity of 100%, moreover b2a3 and b3a3 were predominant. The detecting atypical BCR/ABL mRNA transcripts by real-time quantitative PCR is suitable for the diagnosis of CML negative for P210, P190 and P230 by standard real-time PCR test, and this detection is still the standard and economic method for monitoring minimal residual disease in CML patients with variants of BCR/ABL fusion gene.

Infraspinatus and Teres Minor Tendon Calcification with Severe Pain

Calcific tendinitis is an extremely common shoulder disorder which affects 2.7 to 10.3% of the adult population. However, clear details regarding the formation of calcific tendinitis have only been found recently, and its pathology still under debate. In this report, we present a case of calcific tendinitis affect both the infraspinatus and the teres minor tendon with both X-ray and ultrasound findings. Calcific tendinitis at the infraspinatus is rare (around 8.4% in all rotator cuff calcific tendinitis), and no separate case report on calcific tendinitis has been reported in the teres minor region in the preceding several years, therefore, the rarity of reporting this case was considered.4 We assume that additional case reports can help to further improve our understanding regarding infraspinatus and teres minor tendon calcification.

Interaction of a bioactive pyrazole derivative with calf thymus DNA: Deciphering the mode of binding by multi-spectroscopic and molecular docking investigations

Deoxyribonuclic acid (DNA) is the most relevant intracellular target for a wide variety of anticancer and antibiotic drugs. Elucidating the binding interaction of small bioactive molecules with DNA provides a structural guideline for designing new drugs with improved selectivity and superior clinical efficacy. In the present work interaction of a newly synthesized biologically relevant fluorophore, namely, (E)-1,5-diphenyl-3-styryl-4,5-dihydro-1H-pyrazole (DSDP) with calf thymus DNA (ctDNA) has been investigated vividly through a number of in vitro studies. Noteworthy modifications in the UV–Vis absorption and emission spectra reveal the formation of the probe–ctDNA complex. Several other spectroscopic experiments such as circular dichromism (CD), iodide induced quenching, competitive binding assay with known groove binder probe, 3-hydroxyflavone (3HF), time resolved fluorescence decay measurements, thermometric experiment in connection with the helix melting of ctDNA etc. unequivocally ascertain the groove binding interaction of DSDP with ctDNA. Determination of the thermodynamic parameters through temperature variation study implies the dominant role of hydrophobic interaction in the probe―DNA binding process. Inappreciable change in the CD spectrum of ctDNA with the addition of DSDP suggests that binding of the probe with the DNA does not lead to a significant modification in the DNA conformation. In-silico molecular docking simulation corroborates the experimental findings and depicts that DSDP favorably binds to the minor groove region of the biomacromolecule.

Nuclear Magnetic Resonance Structure of the Human Polyoma JC Virus Agnoprotein.

Agnoprotein is an important regulatory protein of the human polyoma JC virus (JCV) and plays critical roles during the viral replication cycle. It forms highly stable dimers and oligomers through its Leu/Ile/Phe-rich domain, which is important for the stability and function of the protein. We recently resolved the partial 3D structure of this protein by NMR using a synthetic peptide encompassing amino acids Thr17 to Gln52, where the Leu/Ile/Phe- rich region was found to adopt a major alpha-helix conformation spanning amino acids 23-39. Here, we report the resolution of the 3D structure of full-length JCV agnoprotein by NMR, which not only confirmed the existence of the previously reported major α-helix domain at the same position but also revealed the presence of an additional minor α-helix region spanning amino acid residues Leu6 to lys13. The remaining regions of the protein adopt an intrinsically unstructured conformation. J. Cell. Biochem. 118: 3268-3280, 2017. © 2017 Wiley Periodicals, Inc.

Binding of resveratrol to the minor groove of DNA sequences with AATT and TTAA segments induces differential stability

Interactions of a natural stilbene compound, resveratrol with two DNA sequences containing AATT/TTAA segments have been studied. Resveratrol is found to interact with both the sequences. The mode of interaction has been studied using absorption, steady state fluorescence and circular dichroism spectroscopic techniques. UV–visible absorption and fluorescence studies provided the information regarding the binding constants and the stoichiometry of binding, whereas circular dichroism studies depicted the structural changes in DNA upon resveratrol binding. Our results evidenced that, though resveratrol showed similar affinity to both the sequences, the mode of interactions was different. The binding constants of resveratrol to AATT/TTAA sequences were found to be 7.55×105M−1 and 5.42×105M−1 respectively. Spectroscopic data evidenced for a groove binding interaction. Melting studies showed that the binding of resveratrol induces differential stability to the DNA sequences d(CGTTAACG)2 and d(CGAATTCG)2. Fluorescence data showed a stoichiometry of 1:1 for d(CGAATTCG)2-resveratrol complex and 1:4 for d(CGTTAACG)2-resveratrol complex. Molecular docking studies demonstrated that resveratrol binds to the minor groove region of both the sequences to form stable complexes with varied atomic contacts to the DNA bases or backbone. Both the complexes are stabilized by hydrogen bond formation. Our results evidenced that modulation of DNA sequence within the same bases can greatly alter the binding geometry and stability of the complex upon binding to small molecule inhibitor compounds like resveratrol.

Self-Reported Psychosomatic Complaints In Swedish Children, Adolescents, and Young Adults Living in Rural and Urban Areas: An Internet-Based Survey

BackgroundFrequencies in reported psychosomatic illnesses have increased in Sweden among children, adolescents, and young adults. Little is known about demographic differences in self-reported psychosomatic complaints, such as between urban and rural areas, and whether surveys launched on the Internet could be a useful method in sampling such data.ObjectivesThis study examines the connection between psychosomatic illnesses and demographics in Swedish children and youth. The feasibility of using the Internet to gather large amounts of data regarding psychosomatic complaints in this group is another major objective of this study.MethodsA cross-sectional study using 7 validated questions about psychosomatic health, were launched in a controlled way onto a recognized Swedish Internet community site, which targeted users 10 to 24 years of age. The subjects were able to answer the items while they were logged in to their personal domain. The results were analyzed cross-geographically within Sweden.ResultsIn total, we received 100,000 to 130,000 individual answers per question. Subjects of both sexes generally reported significantly higher levels of self-reported psychosomatic complaints in major city areas as compared with minor city/rural areas, even though the differences between the areas were small. For example, 12.00% (4472/37,265) of females in minor regions reported always feeling tense, compared with 13.80% (3156/22,873) of females in major regions (P<.001). In males, the answer pattern was similar, 16.40% (4887/29,801) in major regions versus 15.60% (2712/17,386) in minor regions, (P=.006). Females reported significantly higher frequencies of psychosomatic complaints than males (P<.001).ConclusionsIn subjects aged 10 to 24 years, higher levels of psychosomatic complaints appear to correlate with living in major city areas in comparison with minor city/rural areas. Surveys launched on the Internet could be a useful method in sampling data regarding psychosomatic health for this age group.

Predicting the Warm Forming Behavior of WE43 and AA5086 Alloys

In the present work, we have studied the formability behaviour of two types of magnesium alloys, WE43 hot rolled and WE43 cold rolled by carrying out uniaxial tensile test at elevated temperatures of 350°C to 500°C both experimentally and numerically at a constant strain rate of 10-3s-1. Finite element (FE) model is simulated in ABAQUS/CAE 6.7-6 using coupled temperature-displacement step at higher temperature considering material's property to be isotropic in nature. The effect of temperature on maximum flow stress and major strain at onset of necking is discussed. The true stress-strain behaviour and necking evolution through strain mapping are predicted from FE model and compared with the experimental results. The results show that with increase in temperature, the maximum flow stress decreases and necking delays with increase in limiting major strain for the Magnesium alloys. The work has been extended to predict the forming limit strains of Al 5086 alloy only on the negative minor strain region using M-K (Marciniak and Kuczynski) concept. An FE model based on M-K concept is simulated at 20°C, 150°C and 200°C using coupled temperature-displacement step considering anisotropic sheet material. A groove is created in the middle of the model with an optimizedf value of 0.99, after much iteration. The forming limit strains from such FE simulations are compared with the available experimental data. The results are encouraging providing scope for further improvements in modelling.

Inter-Cell Interference Mitigation Techniques in Long Term Evolution Networks: A Survey

Objectives: This paper gives survey of different techniques and the overview of how the bandwidth is dispensed to the users at the major/edge regions and minor/center regions .It also gives an overview of how the power is dispensed to the users at minor group and major group regions Methods: ICIC has one of the schemes which is based on reusing the frequency called as Fractional Frequency Reuse (FFR). There are different techniques of FFR. They are Partial Frequency Reuse (PFR), Soft Frequency Reuse (SFR) and Soft Fractional Frequency Reuse (SFFR). Findings: In this survey it is found that, the different techniques of FFR gives different throughput and it is also found that SFFR scheme gives the highest throughput when compared with others. Applications: The different techniques of FFR can be used in LTE (Long Term Evolution) based networks. The users will get benefitted based on different power allocations. The real time applications of this technique can be seen in 4G networks.

MOBILITY OF POPULATION IN SUPPORT OF COMPETITIVE ADVANTAGES OF HUMAN CAPITAL

The article discusses the role of mobility of population as a bearer of human potential in shaping the competitive advantages and the accumulation of human capital as the main development resource. It focuses on the problem of polarization in the global market of education, which is based on the concentration of scientific potential and intelligence in minor regions of the planet and its degradation in other regions. The article analyzes the importance of attracting skilled workers to countries based on the Global Competitiveness Index. It reveals the essence of the new "win-win" concept of the circulation of skilled workers. The articledeals with the establishing backlog of Ukraine in attracting highly qualified specialists for the accumulation of human capital. It analyzes the migration losses of Ukraine and identifies prospects of attracting employees with the necessary skills abroad. It is found out that the distribution of migrant workers by occupational groups in employment abroad is significantly different from the structure of professions employed by Ukraine. The article proves the source of human capital loss because of non-recognition in Ukraine the specialties acquired by workers without professional education, which hinders their employment in other sectors, that are specialized according to their formal education at home. It considers the new possibilities of migration transformation towards the creation of the challenges for public policy in connection with the liberalization of foreign trade, harmonization of national legislation with European standards; support the necessary population amount and labor; the formation of orientation on temporary migration (mainly circular) among mobile youth; reintegration of return migrants in the society that has changed over the period of their absence; promoting entrepreneurship of migrants and their families. The article determines the measures to enhance the attractiveness of national labor market to attract human capital through mobility.

Novel e8a2 BCR/ABL Fusion Transcript in Case of a Myeloproliferative Neoplasm.

Main objective of this work was to confirm the occurrence of rare BCR-ABL fusion variant involving the a2 region of the ABL gene and e8 of BCR gene in a patient of Myeloproliferative neoplasm positive for t(9;22) translocation but negative for common major and minor breakpoint cluster regions. A patient with elevated white blood cell count was subjected to classical cytogenetics, FISH as well as RT-PCR testing using commercial kits as well as published primers and in house testing protocol. The translocation event in chromosome 9 and 22 could be successfully detected. BCR/ABL dual color, dual fusion probe generated a classical balanced translocation scenario within the nucleus. In RT-PCR we found an unexpectedly large amplification band around 1700bp, which is consistent with e8a2 transcript. Nine metaphases showed 46,XY,t(9;22)(q34;q11.2)[9] by cytogenetic. A rare e8a2 break point in the BCR-ABL gene in myeloproliferative neoplasm disease detected in India. It also emphasizes the utility of cytogenetic and FISH for primary diagnosis of any neoplasm in blood. Our Patient detected rare BCR-ABL fusion variant e8a2 was on imatinib 400mg since last 3months. After 3months fluorescent in situ analysis and reverse transcriptase pcr analysis showed negative results.

Whole-Genome Comparative Analysis of Two Carbapenem-Resistant ST-258 Klebsiella pneumoniae Strains Isolated during a North-Eastern Ohio Outbreak: Differences within the High Heterogeneity Zones.

Klebsiella pneumoniae has become one of the most dangerous causative agents of hospital infections due to the acquisition of resistance to carbapenems, one of the last resort families of antibiotics. Resistance is usually mediated by carbapenemases coded for by different classes of genes. A prolonged outbreak of carbapenem-resistant K. pneumoniae infections has been recently described in northeastern Ohio. Most strains isolated from patients during this outbreak belong to MLST sequence type 258 (ST258). To understand more about this outbreak two isolates (strains 140 and 677), one of them responsible for a fatal infection, were selected for genome comparison analyses. Whole genome map and sequence comparisons demonstrated that both strains are highly related showing 99% average nucleotide identity. However, the genomes differ at the so-called high heterogeneity zone (HHZ) and other minor regions. This study identifies the potential value of the HHZ as a potential marker for K. pneumoniae clinical and epidemiological studies.

5-Methylcytosine-Rich Heterochromatin in the Indian Muntjac

Two 5-methylcytosine (5-MeC)-rich heterochromatic regions were demonstrated in metaphase chromosomes of the Indian muntjac by indirect immunofluorescence using a monoclonal anti-5-MeC antibody. The metaphases were obtained from diploid and triploid cell lines. A major region is located in the ‘neck' of the 3;X fusion chromosome and can be detected after denaturation of the chromosomal DNA with UV-light irradiation for 1 h. It is located exactly at the border of the X chromosome and the translocated autosome 3. A minor region is found in the centromeric region of the free autosome 3 after denaturing the chromosomal DNA for 3 h or longer. The structure and possible function of the major hypermethylated region as barrier against spreading of the X-inactivation process into the autosome 3 is discussed.