Research Article

Table of Contents | Genet. Mol. Res. 2017 (3)

Displaying 91 - 105 of 115
Research Article

We aimed to investigate the biological role of miR-367 in uveal melanoma cell growth and migration, and the underlying mechanism responsible. Quantitative real-time polymerase chain reaction was performed to evaluate miR-367 expression in uveal melanoma tissue samples and cell lines. A miR-367 mimic, miR-367 inhibitor, and negative control oligonucleotide were transfected into these cells to investigate the function of this microRNA. In addition, the role of PTEN in miR-367-mediated uveal melanoma cell growth and migration was evaluated. miR-367 was significantly upregulated in uveal melanoma cells and tissue samples (both P < 0.01). Its inhibition suppressed the proliferation, cell cycle transition, and migration of such cells, and increased levels had the opposite effect. PTEN was confirmed to be a target gene of miR-367. More importantly, co-transfection with a PTEN construct lacking the 3'-untranslated region mitigated miR-367 mimic-induced promotion of uveal melanoma cell proliferation and migration. In summary, miR-367 was found to be upregulated in this malignancy, and may promote uveal melanoma cell proliferation and migration, at least in part by regulating PTEN.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039067
Research Article

The objectives of this study were to estimate the stability of dry biomass production of elephant grass genotypes under an annual harvest regime, in soil-climatic conditions of the Northern region of the Rio de Janeiro State, Brazil, and to compare methodologies for stability analyses of Yates and Cochran (YC), Plaisted and Peterson (PP), Annicchiarico (ANN), Lin and Binns (LB), Huenh (HU), and Kang and Phan (KP). A randomized block design with 83 treatments and two replicates was adopted. Four annual harvests were performed (2012-2015) and dry matter yield (DMY, t.ha.year) was evaluated. Individual and combined analyses of variance for DMY revealed significant effects for genotypes, harvests, and for the genotype x harvest interaction at the probability levels of 1 and 5%. Genotypes indicated by the YC and PP methods are associated with higher stability and lower DMY. The weighting of KP with YC and PP was highly effective in associating stability with DMY. The LB and ANN methods showed strong agreement with each other and produced similar classifications as to phenotypic stability, and so we recommend using one or the other. Genotypes Elefante Cachoeiro do Itapemirim, Cuba-116, Taiwan A-46, P241 Piracicaba, Taiwan A-144, Cameroon - Piracicaba, 10 AD IRI, Guaçu/I,Z,2, Mineirão IPEACO, Taiwan A-121, IJ7125 cv EMPASC308, 903-77, Mole de Volta Grande, and Porto Rico 534-B showed high stability and DMY, standing out as promising genotypes for the soil-climatic conditions of the Northern region of the Rio de Janeiro State. The methodologies based on ANOVA and non-parametric analyses were complementary and increased reliability in the recommendation of genotypes.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039041
Research Article

The forkhead box F2 (Foxf2) gene suppresses epithelial-mesenchymal transition via the modulation of transcription of zinc finger E-box-binding homeobox 1 (Zeb1) and epithelial (E)-cadherin, thereby inhibiting tumor metastasis. Additionally, the specific binding of microRNA (miR)-200c to Foxf2 mRNA impedes metastatic pulmonary cancer. However, the role of miR-200c in breast cancer is still unknown. Therefore, in this study, miR-200c mimics were transfected into the highly metastatic breast cancer cell line MDA-MB-231. Their invasion and migration abilities were observed by scratch and transwell migration assays. Real-time PCR was used to detect mRNA levels of Foxf2, Zeb1, and E-cadherin, whereas Foxf2 protein level was determined by western blot analysis. Our results showed that, compared to the control group, miR-200c inhibited the migration or invasion of MDA-MB-231 cells. Real-time PCR and western blot analysis exhibited significant decreases in Foxf2 expression in the presence of miR-200c, along with a decrease in Zeb1 and increase in E-cadherin mRNA expressions. Thus, our preliminary data demonstrated that miR-200c could inhibit the metastasis of breast cancer cells by downregulating Foxf2 expression, providing leads for the discovery of a novel breast cancer treatment.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16038971
Research Article

This study aimed to evaluate the supplementation levels of an exogenous alpha-amylase in broilers diets and compare two indicators in determining the diets energy. The experiment was divided into two parallel evaluations, being one of performance and the other of metabolism. In performance assay, 1,700 one-day-old Cobb-500 male chicks were used. The animals were distributed in 50 experimental plots and evaluated five treatments with ten replicates in a completely randomized design (CRD). The treatments were: a positive control (PC), a negative control (NC) and three alpha-amylase supplementation levels 200, 400 and 600 g/t, and the NC was formulated with 50 and 90 kcal of energy reduction in relation to the PC to the phases from 1 to 21 days and from 22 to 42 days, respectively. In the metabolism assay were used 240 animals, 150 birds for stage from 14 to 21 days and 90 birds to stage from 35 to 42 days of age and the treatments were the same as the performance assay, with six replicates per treatment in CRD. All diets of metabolism test contained the digestibility indicators Lipe (eucalyptus purified lignin) and chromic oxide (CrO), in concentrations of 0.05 and 1.0%, respectively. In the period from 1 to 21 days old, no significant differences were observed in weight gain (WG) (P > 0.05), however, feed intake (FI) was found higher by using 200 ppm of enzyme (P < 0.05) and better feed conversion (FC) with the PC (P < 0.05). From 22 to 42 days, no significant differences were observed on the WG (P > 0.05), but were observed lower FI and better FC to PC treatment (P < 0.05). In the period from 1 to 42 days of age, significant differences were also not observed on the WG (P > 0.05), but there was lower FI and better FC for the PC treatment (P < 0.05). The AMEn (apparent metabolizable energy corrected for nitrogen balance), determined using the total collection, reaffirmed the values ​​calculated for the PC and NC with intermediate data obtained from the enzyme use (200, 400 and 600 ppm). Comparing the total collection using Lipe and CrO, a correlation was observed only for the PC results, that were always higher, and for the NC results, that were lower for the three methodologies. For IDE (ileal digestible energy), determined by CrO significant differences were observed (P < 0.05) and it presented higher values in the PC treatment and lower values in the NC. In IDE determining by Lipe, significant was observed (P < 0.05), showing higher value in the PC treatment. It is concluded that in the metabolism assessments, exogenous alpha-amylase tested was effective in increasing the metabolizable energy (ME), however, not enough to be equivalent to the ME of PC treatment, a result that was better expressed in the total collection, AMEn by Lipe and IDE by Lipe methodologies. The performance results reflect the superiority of PC treatment, pointing enzyme limitation to supply the energy deficit practiced in this work.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16038766
Research Article

The Malaysian giant prawn is among the most commonly cultured species of the genus Macrobrachium. Stocks of giant prawns from four rivers in Peninsular Malaysia have been used for aquaculture over the past 25 years, which has led to repeated harvesting, restocking, and transplantation between rivers. Consequently, a stock improvement program is now important to avoid the depletion of wild stocks and the loss of genetic diversity. However, the success of such an improvement program depends on our knowledge of the genetic variation of these base populations. The aim of the current study was to estimate genetic variation and differentiation of these riverine sources using novel expressed sequence tag-microsatellite (EST-SSR) markers, which not only are informative on genetic diversity but also provide information on immune and metabolic traits. Our findings indicated that the tested stocks have inbreeding depression due to a significant deficiency in heterozygotes, and F was estimated as 0.15538 to 0.31938. An F-statistics analysis suggested that the stocks are composed of one large panmictic population. Among the four locations, stocks from Johor, in the southern region of the peninsular, showed higher allelic and genetic diversity than the other stocks. To overcome inbreeding problems, the Johor population could be used as a base population in a stock improvement program by crossing to the other populations. The study demonstrated that EST-SSR markers can be incorporated in future marker assisted breeding to aid the proper management of the stocks by breeders and stakeholders in Malaysia.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16035685
Research Article

A disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4) can effectively degrade articular cartilage matrix proteoglycan and damage the intervertebral disc of spinal tuberculosis patients, resulting in deterioration of the physical properties of articular cartilage. Transforming growth factor β activated kinase 1 (TAK1) is similar to vascular cell adhesion molecule 1 (VCAM-1) and closely related to a variety of pathophysiological processes. This study intended to explore the expression of ADAMTS-4, VCAM-1, and TAK1 in cartilage tissue obtained from spinal tuberculosis patients and their inter-relationships, aiming to provide new treatment approaches for spinal tuberculosis. Patients with spinal tuberculosis (N = 60) from the department of orthopedics and patients with traumatic spinal fracture (N = 60, controls) were recruited for the study. ADAMTS-4, VCAM-1, and TAK1 expression was detected by immunohistochemistry. SPSS 19.0 software was used for data processing and analysis. The score values of ADAMTS-4, TAK1, and VCAM-1 were 1.45 ± 0.10, 1.33 ± 0.09, and 1.54 ± 0.11, respectively, which were significantly higher than those in normal controls (P < 0.05). ADAMTS-4 showed positive correlation with VCAM-1 and TAK1. ADAMTS-4, TAK1, and VCAM-1 expressions increased in spinal tuberculosis patients. They could provide clinical reference for spinal tuberculosis diagnosis and new treatment strategies can be devised by focusing on their positive correlation.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16038961
Research Article

Genotypes x environment (G x E) interaction consists of different behavior of genotypes cultivated in different environments. This interaction occurs due to the performance variation of each genotype in different environments. To reduce the effect of the interaction in soybean crops, some studies have been reported in the literature to study their adaptability and stability. However, these studies are still scarce in Minas Gerais State. Thus, the aim of this study was to verify the adaptability and stability of soybean cultivars and identify the cultivars that contribute least to the G x E interaction in Minas Gerais. Six soybean cultivars were evaluated in 9 different environments. The plots were composed of 4 rows of 5 m with a spacing of 0.5 m between rows, and only the two central rows were harvested. The inoculation with Bradyrhizobium japonicum was performed through sowing furrow. The fertilization followed the recommendations of the Soil Fertility Commission of Minas Gerais. Grain yield was evaluated in kg/ha after conversion to 13% moisture. After individual analysis, the joint analysis was performed by grouping the phenotypic means by the Scott and Knott (1974) test. Wricke's ecovalence methodologies and the Annicchiarico confidence index were applied for the adaptability and stability analysis. The interaction was decomposed into a simple and a complex part. The cultivars BRSMG 820RR and BRSMG 760SRR have wide adaptability and stability. The first one presents a better index of confidence and a small contribution to the interaction.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039730
Research Article

Genotypes x environment (G x E) interaction consists of different behavior of genotypes cultivated in different environments. This interaction occurs due to the performance variation of each genotype in different environments. To reduce the effect of the interaction in soybean crops, some studies have been reported in the literature to study their adaptability and stability. However, these studies are still scarce in Minas Gerais State. Thus, the aim of this study was to verify the adaptability and stability of soybean cultivars and identify the cultivars that contribute least to the G x E interaction in Minas Gerais. Six soybean cultivars were evaluated in 9 different environments. The plots were composed of 4 rows of 5 m with a spacing of 0.5 m between rows, and only the two central rows were harvested. The inoculation with Bradyrhizobium japonicum was performed through sowing furrow. The fertilization followed the recommendations of the Soil Fertility Commission of Minas Gerais. Grain yield was evaluated in kg/ha after conversion to 13% moisture. After individual analysis, the joint analysis was performed by grouping the phenotypic means by the Scott and Knott (1974) test. Wricke's ecovalence methodologies and the Annicchiarico confidence index were applied for the adaptability and stability analysis. The interaction was decomposed into a simple and a complex part. The cultivars BRSMG 820RR and BRSMG 760SRR have wide adaptability and stability. The first one presents a better index of confidence and a small contribution to the interaction.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039730
Research Article

Genotypes x environment (G x E) interaction consists of different behavior of genotypes cultivated in different environments. This interaction occurs due to the performance variation of each genotype in different environments. To reduce the effect of the interaction in soybean crops, some studies have been reported in the literature to study their adaptability and stability. However, these studies are still scarce in Minas Gerais State. Thus, the aim of this study was to verify the adaptability and stability of soybean cultivars and identify the cultivars that contribute least to the G x E interaction in Minas Gerais. Six soybean cultivars were evaluated in 9 different environments. The plots were composed of 4 rows of 5 m with a spacing of 0.5 m between rows, and only the two central rows were harvested. The inoculation with Bradyrhizobium japonicum was performed through sowing furrow. The fertilization followed the recommendations of the Soil Fertility Commission of Minas Gerais. Grain yield was evaluated in kg/ha after conversion to 13% moisture. After individual analysis, the joint analysis was performed by grouping the phenotypic means by the Scott and Knott (1974) test. Wricke's ecovalence methodologies and the Annicchiarico confidence index were applied for the adaptability and stability analysis. The interaction was decomposed into a simple and a complex part. The cultivars BRSMG 820RR and BRSMG 760SRR have wide adaptability and stability. The first one presents a better index of confidence and a small contribution to the interaction.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039730
Research Article

Anthracnose is among the major diseases of the Capsicum culture. It is caused by different species of the genus Colletotrichum, which may result in major damages to the cultivation of this genus. Studies aiming to search for cultivars resistant to diseases are essential to reduce financial and agricultural losses. The objective of this study was to evaluate the correlation between the variables analyzed to select Capsicum genotypes resistant to the fungus Colletotrichum gloeosporioides. The experimental design was completely randomized blocks with three replications, 88 treatments, four ripe fruits, and four unripe fruits per replication. Accessions of Capsicum from the Germplasm Active Bank of Universidade do Estado de Mato Grosso (UNEMAT) were evaluated as for resistance to the fungus. Fruits were collected from each plot and taken to the laboratory for disinfestation. A lesion was performed in the middle region of the fruit using a sterile needle, where a spore suspension drop, adjusted to 10 spores/mL, was deposited. An ultrapure water drop was deposited into control fruits. The fruits were placed in humid chambers, and the evaluation was performed by measuring the diameter and the length of lesions using a caliper for 11 days. After data were obtained, analyses of variance, correlation, and path analysis were performed using the GENES software and R. According to the likelihood-ratio test, the effects of genotypes (G), fruit stage (F), and its interaction (G x F) were significant (P < 0.05). There were differences between the magnitudes of genotype correlations according to fruit stage. Different variables must be taken into account for an indirect selection in this culture in function of fruit stage since the variable AUDPC is an important criterion for selecting resistant accessions. We found through the path analysis that the variables DULRD and DULRL exerted the greatest effects on AUDPC.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039720
Research Article

Anthracnose is among the major diseases of the Capsicum culture. It is caused by different species of the genus Colletotrichum, which may result in major damages to the cultivation of this genus. Studies aiming to search for cultivars resistant to diseases are essential to reduce financial and agricultural losses. The objective of this study was to evaluate the correlation between the variables analyzed to select Capsicum genotypes resistant to the fungus Colletotrichum gloeosporioides. The experimental design was completely randomized blocks with three replications, 88 treatments, four ripe fruits, and four unripe fruits per replication. Accessions of Capsicum from the Germplasm Active Bank of Universidade do Estado de Mato Grosso (UNEMAT) were evaluated as for resistance to the fungus. Fruits were collected from each plot and taken to the laboratory for disinfestation. A lesion was performed in the middle region of the fruit using a sterile needle, where a spore suspension drop, adjusted to 10 spores/mL, was deposited. An ultrapure water drop was deposited into control fruits. The fruits were placed in humid chambers, and the evaluation was performed by measuring the diameter and the length of lesions using a caliper for 11 days. After data were obtained, analyses of variance, correlation, and path analysis were performed using the GENES software and R. According to the likelihood-ratio test, the effects of genotypes (G), fruit stage (F), and its interaction (G x F) were significant (P < 0.05). There were differences between the magnitudes of genotype correlations according to fruit stage. Different variables must be taken into account for an indirect selection in this culture in function of fruit stage since the variable AUDPC is an important criterion for selecting resistant accessions. We found through the path analysis that the variables DULRD and DULRL exerted the greatest effects on AUDPC.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039720
Research Article

A disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS-4) can effectively degrade articular cartilage matrix proteoglycan and damage the intervertebral disc of spinal tuberculosis patients, resulting in deterioration of the physical properties of articular cartilage. Transforming growth factor β activated kinase 1 (TAK1) is similar to vascular cell adhesion molecule 1 (VCAM-1) and closely related to a variety of pathophysiological processes. This study intended to explore the expression of ADAMTS-4, VCAM-1, and TAK1 in cartilage tissue obtained from spinal tuberculosis patients and their inter-relationships, aiming to provide new treatment approaches for spinal tuberculosis. Patients with spinal tuberculosis (N = 60) from the department of orthopedics and patients with traumatic spinal fracture (N = 60, controls) were recruited for the study. ADAMTS-4, VCAM-1, and TAK1 expression was detected by immunohistochemistry. SPSS 19.0 software was used for data processing and analysis. The score values of ADAMTS-4, TAK1, and VCAM-1 were 1.45 ± 0.10, 1.33 ± 0.09, and 1.54 ± 0.11, respectively, which were significantly higher than those in normal controls (P < 0.05). ADAMTS-4 showed positive correlation with VCAM-1 and TAK1. ADAMTS-4, TAK1, and VCAM-1 expressions increased in spinal tuberculosis patients. They could provide clinical reference for spinal tuberculosis diagnosis and new treatment strategies can be devised by focusing on their positive correlation.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16038961
Research Article

The lower northern part of Thailand contains various genetically diverse ethnic populations. The sequences of the mitochondrial DNA hypervariable region were studied in three ethnic populations inhabiting Phitsanulok Province. One hundred and nine nucleotide sequences - 53, 29, and 27 from Hmongs (Hill tribe), Lao Songs, and Thai-Siams, respectively - were collected. The haplotypes were generated from 1130 nucleotides of the entire control region. Eighty-six haplotypes were found in the three ethnic populations, and no shared haplotypes were found between populations. Point heteroplasmy was noted at position 311 (C→Y). Haplotypes with ACAC-insertion at position 512 were observed in immigrant individuals from the Lao Song population. The Thai-Siam population showed higher genetic diversity than the other populations. The Hmong and Lao Song populations showed less genetic diversity than those living in their original area. The neutrality testing suggested that the result might be influenced by genetic drift.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039687
Research Article

The present study was carried out in a native Brazil nut tree population (Bertholletia excelsa Bonpl., Lecythidaceae) to assess its genetic diversity and structure. Ten microsatellite markers were used to genotype 198 adult trees (B. excelsa). The population presented high genetic diversity and inbreeding absence rates. The empirical Bayesian method showed three distinct groups in the structure of this population. Molecular analysis of variance showed 98% variability within groups, and 2% between groups. The genetic divergence (F) indicated little difference between groups; thus, suggesting efficient gene flow between the analyzed B. excelsa adult trees.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039702
Research Article

A role for estrogen in the etiology of breast and ovarian cancers has been suggested; therefore, genetic polymorphisms in steroid metabolism genes could be involved in the carcinogenesis of these tumors. We have aimed to investigate the role of GSTP1 and CYP17 polymorphisms and their correlation with MSI (microsatellite instability) and LOH (loss of heterozygosity) in AR, ERβ and CYP19 genes in women from Espírito Santo State, Brazil. The study population consisted of 107 female breast and 24 ovarian tumors. GSTP1 and CYP17 polymorphisms were detected by polymerase chain reaction (PCR) amplification followed by restriction fragment length polymorphism (RFLP) analysis while MSI and LOH were analyzed by PCR. GSTP1 and CYP17 polymorphisms alone were not associated with an increased risk for breast or ovarian tumors. However, when combined with MSI/LOH in AR, ERβ and CYP19 genes, we were able to detect significant associations with the GSTP1 wild-type genotype in PR (progesterone receptor) negative breast cancers or the CYP17 wild-type genotype in ER (estrogen receptor) and PR-negative breast tumors. No associations with ovarian tumors were detected. Our results suggest that wild-type GSTP1 or CYP17 genes when combined with LOH/MSI in steroid metabolism genes may play a role in ER and/or PR negative breast cancers. These data support the hypothesis that genes related to steroid metabolism are important in the characterization of breast cancer and that the analysis of single polymorphisms may not be sufficient.

Genet. Mol. Res. 2017
DOI: 10.4238/gmr16039691

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