Research Article

Table of Contents | Genet. Mol. Res. 2017 (4)

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Research Article

 Precocious soybean cultivars enable crop rotation and
low pressure to the biotic and abiotic factors. This study aimed to
determinate the inheritance of characters related to precocity and
agronomic characters in soybean segregating population, coming from
contrasting genitors to cycle. The experiment was conducted at a
greenhouse located at Capim Branco farm, belonging to Federal
University of Uberlândia. The MG/BR 46 Conquista and UFUS 6901
cultivars were used to bi-parental crossing and acquirement of F1 and
F2 populations. The individuals of the genitors, F1 and F2 populations
were evaluated to eight characters and the phenotypic, genetic and
environmental variances, broad-sense heritability, average degree of
dominance and gene number were determined. The average for number
of days for flowering and number of days to maturity in the F1
generation was 40.26 and 103.52 days and in the F2 generation was
37.85 and 105.88 days, respectively, which fits into the classification of
semi-precocious. The heritability varied from 2.36% for number of
grains per pod – controlled by 160 genes, to 85.39% for number of days
for maturing – controlled by two genes. On the F2 population, the
existence of transgressed segregates was found, except for number of
days to maturity, number of nodes and intersection height of the first
pod. It was concluded that the genetic variance and heritability of the acquirement of
selection of superior individuals in terms of precocity, height, number of
nodes and compounds of productivity, being promising genotypes in the
soybean breeding program.
 

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039842
Research Article

 The Klf6 gene is a tumor suppressor gene belonging to
the family of the Klf gene and closely associated with tumor formation.
Recently, super enhancers(SEs) have been shown to play a particularly
important role in regulating cell identity and are attempting to evaluate
the in vivo function of SEs. But direct functional evidence of SE
associated with Klf6 is lacking. Using genomic editing technology, we
have attempted to identify super enhancer associated with the expression
of Klf6 and clarify its function in human hepatoma (HepG2) cells. As a
result, it identified the Klf6 related SE and demonstrated that the Klf6-
related SE is responsible for more than 80% of Klf6 gene expression. It
also revealed the hierarchical structure of the Klf-6 related SE and the
function of individual enhancers. Our results provide the functional
significance of the super enhancer in understanding the transcriptional
regulation mechanism of Klf6.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039841
Research Article

 Antibiotic-induced disruption of the intestinal microbiota
has serious consequences for human physiology. We conducted a
microbial dysbiosis animal model to study and directly provide evidence
for microbial dysbiosis contribution to malignant melanoma animal
model. Females C57BL/6 wild-type (WT) mice injected with a limiting
threshold number of B16/F1 melanoma cells and the tumor development
was observed with and without probiotics treatment. Treatment with
probiotic could restore microbiota diversity and indirectly effect the
tumor incidence. In microbial dysbiosis group tumor incidence was
83.33%. Also, the expression level of inflammatory proteins NFkB-p65;
IL-6; and STAT-3 were remarkably enhanced in microbial dysbiosis
group compared to probiotic treated group. Altogether, our findings
demonstrated that microbial dysbiosis can strike the balance between
immunity and tumorigenesis, and increase the incidence of malignant
melanoma in mice. Probiotic treatment significantly reduced tumor
incidence.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039840
Research Article

Brazil nut tree is a species of economic importance for
the Amazon region, known for the commercialization of its almonds.
The objective of this work was to study the genetic diversity among halfsib
progenies from different Brazil nut trees present in native forest in
the municipality of Itaúba, MT, belonging to the Brazilian Amazon. In a
native forest area of nine hectares, fruits of nine parent trees, randomly
selected in the plot, were collected. The seeds were planted at
greenhouse and they were named, according to their origin, identifying
seed tree and fruit. After the seed germination and initial development
of the seedlings, leaves were collected for DNA extraction and analyzed
with microsatellite molecular markers. It was performed analysis of
molecular variance and cluster analysis of progenies and seed trees.
There is greater genetic diversity between families than among
progenies from the same family. The clustering of progenies from
different families in the same group can be explained by the low
dissimilarity between the seed trees. Among the loci analyzed in this
study, eight were informative for evaluations of genetic diversity in
Brazil nut, except BET12 and BET16 loci.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039839
Research Article

Onion (Allium cepa L.) is the third most valuable
vegetable crop worldwide and the third most produced in Brazil. The
knowledge of the nature and the value of the relationship between the
traits of interest is very important, since it is possible to select a main
trait, with low heritability or difficult measurement, obtaining higher
genetic gain faster than with direct selection. The aim of this work was
to estimate the genotypic, phenotypic and environmental correlations
between the characteristics of the plant, the bulb and seed production in
onion segregating populations obtained from different breeding
methods. Most of the estimations of the genetic correlations (rg) were
higher than the phenotypic (rp) and the environmental (re) ones. The
genotypic, phenotypic and environmental correlation coefficients for
plant and bulb characteristics were low, except for plant vigour and its
components (plant height, neck diameter and plant architecture). The
phenotypic and genotypic correlations revealed an association between
the traits related to the plant vigour in onion, nevertheless, it was not
reflected in the bulb and seed production, in the two years evaluated. The
traits “Seed setting” and “Resistance in Alternaria” can be useful in the
indirect selection for “Seed mass per umbel.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039838
Research Article

Low-cost and low-density (LCD) DNA arrays offer an
easy way to detect resistance as minimal laboratory instrumentation is
needed. Nucleic acid-based amplification tests allow the rapid detection
of Mycobacterium tuberculosis. Recently, a real-time PCR assay for M.
tuberculosis complex was introduced. Real-Time PCR and DNAmicroarray
techniques were compared with the classical methods of
Ziehl-Neelsen (ZN) staining and culturing. Regarding to the standard
culture method, 80 positive individuals were identified out of 140 urine
samples. RT- PCR showed 96.3% sensitivity and 96.7% specificity with
Mycobacterial tuberculosis complex (MTB) (n=10) and nontuberculous
mycobacteria (NTM) (n=70). The DNA-microarray analysis exhibited
100% sensitivity and specificity. One species belonging to (MTB) was
identified as M. tuberculosis and positively represented by 12.5%
(n=10). Five species belonging to nontuberculous mycobacteria (NTM)
were identified and represented as M. kansasii 37.5% (n=30), M.
celatum 21.25% (n=17), M. gordonae 11.25% (n=9), M. chelonae 10%
(n=8), and M. phlei 7.5% (n=6). The results recommend the use of our
simple and rapid PCR technique for early diagnosis of mycobacteria’s. 
Also, the fast LCD-microarray protocol is very useful for species
identification and differentiation between MTB and NTM.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039837
Research Article

The TCTP (Translationally controlled tumor protein)
is a highly conserved protein family found in eukaryotic organisms,
which has been associated to the performance of several functions at
the cellular level, as those related to growth, development, and
responses against environmental stresses. However, studies on TCTP
from plants are still incipient. Cassava (Manihot esculenta Crantz) is
a crop of great socioeconomic importance for millions of people in
the world, especially because of its high energy content. Previous
studies reported the identification of a cassava TCTP (MeTCTP)
with potential roles in storage root formation and salt stress response.
Here, our main goal was to increase the understanding about the roles
of MeTCTP in response to abiotic stresses. We verified that the
overexpression of recombinant MeTCTP in Escherichia coli
increased tolerance of bacterial cells to heat stress. In addition, the
recombinant MeTCTP was purified by nickel affinity and evaluated
by chaperone activity assays, which showed its ability to preserve the
function of NdeI restriction enzyme under thermal denaturation 
 
conditions. Results presented here provided insights into the
relationship of MeTCTP with responses of plants to abiotic stresses.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039835
Research Article

Background: Protein Z (PZ) is a vitamin K-dependent
plasma glycoprotein, it plays a key role in the physiologic inhibition
of coagulation by acting as a cofactor in the inactivation of factor Xa.
The relationship between PZ gene polymorphisms and pregnancy
loss is controversial. To address this, we investigate the association
between PZ (rs3024718, rs3024719, rs3024731, rs3024778 and
rs3024772) single nucleotide polymorphisms (SNPs) and idiopathic
recurrent pregnancy loss (IRPL) in in women from Tabuk region
(North western region, Saudi Arabia).
Methods: A case control study, including Sixty-three women with at
least three unexplained recurrent pregnancy loss (RPL) were selected
and matched with seventy-eight healthy and fertile women
(controls). SNP genotyping was carried out by allelic discrimination
using real time PCR.
Results: The outcome was that the minor allele frequencies (MAF)
are 0.23 vs 0.20, 0.24 vs 0.19, 0.02 vs 0.02, 0.26 vs 0.21, and 0.02 vs
0.01 respectively for rs3024718 A/G, rs024719 G/A, rs3024778
G/A, rs3024731 G/A, and rs3024772 A/G PZ polymorphisms. The
genotype distribution was similar between women with RPL and
control women (P>0.05) and none of the tested SNPs were associated
with RPL under co-dominant, dominant, or recessive genetic models.
The lack of association was also confirmed by haplotypes analysis
showing an absence of RPL risk with the constructed haplotypes.
Conclusion: It was concluded that the studied PZ polymorphisms are
not associated with the risk of recurrent pregnancy loss in the studied
population. 

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039834
Research Article

Purpose: To explore the mechanism of You-Gui-Wan
(YGW) in improving endometrial receptivity in mouse model of
controlled ovarian hyperstimulation (COH).
Methods: Sixty female mice with two consecutive oestrous cycles
were randomly divided into COH model group (n = 50) and normal
control group (n = 10). The COH model was created by using
GnRH-a + PMSG + HCG and further randomly divided into 5
subgroups of simple model, YGW high-dose, YGW medium-dose,
YGW low-dose and aspirin control, 10 in each subgroup. YGW or
aspirin or saline was administrated to the mice by gavage once a day
for 11 days. Afterwards, blood and uterine samples were collected
for examination of serum levels of estradiol (E2) and progesterone
(P4), endometrial thickness and number of blood vessels as well as
expressions of angiogenic factors of VEGF, Ang-1, Ang-2 and Tie2
at transcription and translation levels by using H& E staining,
ELISA, real-time qRT-PCR, and immunohistochemistry,
respectively.
Results: COH per se increased the serum concentrations of E2 and
P4,but decreased the number of endometrial blood vessels and the
protein expressions of the angiogenic factors when compared to the
normal control group (P< 0.05 or 0.01). YGW increased the mRNA
expressions of all angiogenic factors and restored the effect of COH
on number of endometrial blood vessels (P< 0.01), and the protein
expressions of the angiogenic factors when compared to the COH
simple model group (P< 0.05 or 0.01). In comparison, aspirin could
only restore the number of endometrial blood vessels and the protein
expressions of Ang-2 and Tie-2 (P< 0.05 or 0.01).
Conclusions: YGW can restore the side effect of COH on number
of endometrial blood vessels and the expressions of angiogenic
factors, thus improve endometrial receptivity in COH mice.
 

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039833
Research Article

The interaction between the ovine prion protein gene
(PRNP) and Doppel (PRND) gene polymorphisms is essential for
understanding the role of prion proteins in scrapie. In this study, the
blood genomic DNA samples of 111 Gansu Alpine Merino sheep
were used to define the PRNP alleles and the haplotypes of PRND
by PCR-SSCP (single-strand conformation polymorphism) and
PCR. The results shown that the frequency of PRNP genotype
ARQ/ARQ and ARR/ARR has a certain advantage, especially the
ARQ/ARQ (46%). In addition, PRND 26 haplotypes and 29
polymorphisms sites were firstly defined, and the dominant
polymorphic sites are at codons 28, 51 and 158. The frequency of the
haplotype GTC of PRND is 98%, the ARQ/ARQ has great advantage 
in the PRNP genotype. These results may be helpful in future studies
of the relationship between PRND with PRNP in prion diseases. 

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039832
Research Article

Prediabetes is considered to be an at a risk state, with
an annualized 5-10% conversion rate of diabetes and the similiar
proportion converting back to normoglycaemia. The global
incidence of prediabetes is increasing and will reach to 471 million
patients in 2030. The elevation of blood sugar is a continuum and
hence prediabetes cannot be considered an entirely benign status and
present insulin resistance (IR) and β-cell dysfunction, so that the
development of prediabetic complications and vascular endothelial
dysfunction. Therefore, it is crucial to early recognition and
treatment of prediabetic individuals. For prediabetic individuals,
lifestyle modification is the cornerstone of diabetes prevention with
evidence of a 40-70% relative risk reduction. In the review, we
include the possible mechanism of IR-induced vascular endothelial
dysfunction, and exercise intervention improving the vascular
endothelial dysfunction. These results showed clearly positive
effects of physical exercise intervention and provided a direct
clinical evidence of the effectiveness of this approach for preventing
early diabetes

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039830
Research Article

Toll-like receptor 4 (TLR4), one of the key immune
system effectors, plays a main role in immune recognition of cervical
cancer. Micro-RNAs are involved in regulation of multiple important
genes in the progression of cervical cancer. A case-control study of
592 people was conducted from Yun’an County, Yunfu City,
Guangdong Province, China. Cervical fall off epithelia were
collected to detect human papilloma virus (HPV), followed by Thin
Prep cytology test (TCT). Moreover, extraction of DNA from
peripheral blood were performed for genotyping from the 296
patients and another 296 age-matched healthy control subjects.
Logistic regression was used to determine the risk genotypes for
susceptibility to cervical precancerous lesion, and multifactor
dimensionality reduction (MDR) was further employed to
preliminarily investigate the gene-environment interaction on risk of
cervical precancerous lesion. Gene expression of miRNA-140 from
serum was done by real-time PCR. We investigated whether target
sites of TLR4 gene polytheisms (rs11536896 T>C, rs7873784 G>C)
of miR-140 were associated with cervical precancerous lesion risk.
The alleles C>A of SNP rs11536896 were significantly different in
ASCUS in comparison of case group and control group with HPV
infection. The presence of the allele C was associated with a higher
risk of developing ASCUS lesion in HPV negative women (OR:
1.75, 95%CI:1.20-2.54, p = 0.003). There was statistically significant
difference between the expression of miRNA 140 and the
susceptibility to cervical precancerous lesion, in which there is
down-regulation of the miRNA-140 in case group (T=6.73,
P=0.007). Gene-environment interaction analysis by MDR software
revealed an association among rs7873784 and hrHPV infection and
more types of infected HPV (p < 0.0001, OR: 25.48; 95%Cl: 5.20-
124.84). Collectively, these results suggested that rs11536896 and
rs787378 from TLR4 gene were associated with risks of cervical
precancerous lesion. Thus, this miRNA-140 and SNPs(
rs11536896rs787378) of TLR4 gene could be considered as a
potential molecular mechanism and biomarker for detecting and
diagnosing cervical cancer in early time.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039831
Research Article

Anthocyanins are flavonoids compounds that have a great therapeutic potential acting as antioxidants, anti-inflammatory and anti-carcinogenic agents. They are consumed through food, pharmaceutical and cosmetics industries as a natural alternative to synthetic dyes. However, anthocyanins are unstable depending on pH, temperature, light and oxygen variations, which led to the production of its encapsulation in nanoparticles as an alternative to increase its stability and protect them from chemical degradation for human consumption. Due to the fact that many of the beneficial properties of nanoparticles could also be harmful to human health, and the need of prior toxicological evaluation of any medicinal product, this research aims to evaluate the cytotoxic and genotoxic effects of anthocyanins-loaded Eudragit® L100 nanoparticles (AN). A range of AN doses were tested with a preliminary MTT assay, which allowed selecting five concentrations for comet and micronucleus assays: 0.2, 2, 20, 125 and 250 µg/mL, respectively, on human peripheral blood mononuclear cells (PBMC) (cells without liver metabolizing enzymes) and human hepatoma cell line HepG2 (with liver metabolizing enzymes). Results showed absence of cytotoxic effect in MTT test on both cell types after 24 h of exposure. However, the nuclear division index in PBMC cells assessed by micronucleus test indicates a decrease in the cell division, after 28 h of exposure. Genotoxicity analysis showed that AN did not produce significant genotoxic effects detected in comet assay, in either cell type. However, the micronucleus test on HepG2 cells showed that at concentration of 2 µg/mL and higher, AN produced clastogenic/aneugenic effects. Under our experimental conditions and limitations, the observed cytotoxic and genotoxic effects, not previously reported in the literature, may be due to anthocyanin metabolization and related to its concentrations and time of exposure

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039875
Research Article

Euterpe precatoria, popularly known as “açaí-do-amazonas”, has agronomic, technological and economic potential and has been gaining prominence with the commercialization of its fruits. The production of this açaí-do-amazonas is based on extractivism and management practices have been recommended for sustainable collection. To recommend sustainable rates of fruit collection, it is essential to have information on the genetic variability of natural populations to monitor the recruitment and population dynamics. The objective of this study was to evaluate the transferability of microsatellite loci of E. edulis species to E. precatoria to confirm the applicability of these markers in genetic studies. Eighteen microsatellite loci developed for E. edulis were analyzed using 20 individuals from two natural populations (Formoso and Novo Segredo) of E. precatoria collected at Feijó, State of Acre, Brazil. All loci (100%) were amplified, and of these, nine (50%) were polymorphic. A total of 29 alleles were found in the Novo Segredo population, and 27 alleles were found in the Formoso population, ranging from two to five alleles per locus, with a mean of three alleles per locus. The expected heterozygosity in the Formoso population varied from 0.100 to 0.668, with an average of 0.421. In the Novo Segredo population, a variation between 0.100 and 0.710, with a mean of 0.418, was found. The observed heterozygosity values ranged from 0.100 to 0.800, with averages of 0.333 and 0.267 for Formoso and Novo Segredo, respectively. This set of markers will support further studies on the molecular characterization of the natural populations of E. precatoria and assist with the recommendation of sustainable management practices and strategies for the conservation and genetic improvement of this species.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039825
Research Article

A study of intestinal helminthic parasites of domestic chickens was carried out in Tabriz city, Iran during 2016. One hundred (100) gastrointestinal samples were examined in necropsied chickens microscopically using the lactophenol and carmine-acid staining technique. The result showed a high prevalence of intestinal parasitic infection. Out of 100 domestic chicken, 37 (37%) were infected by the parasite. Nematodes had the highest prevalence 25 (68%), followed by the cestodes with the prevalence of 12 (32%) in the parasitic infested birds. Nematodes especially Ascaridi agalli 9 (16 %) and Heterakis galinarum 16 (64 %) were most prevalent in the nematode infested birds. Raillietina tetragona 12 (100%) was found to be the highest in occurrence among the cestodes infested birds. No trematode was encountered among the infested birds. The average parasite burden per chicken was found to be 5.7 and majority of the species were restricted to the small and large intestine. 14 (38 %) cases of mixed infections were encountered out of the infested poultry. The study reveals that there was a high prevalence of intestinal parasites of domestic chicken in the study area. This calls for improved management and disease control to enhance their potential.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039824

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