Fish eggs are often excluded from identification analysis since at this stage of development there are few morphological characters. The correct identification of eggs can provide important ...information about spawning areas of species. The current work aimed to identify fish eggs in the Tibagi and Cinzas Rivers using the DNA barcode to obtain information on richness and diversity, adding to the existing data in the area. Of the 928 sequences analyzed using the BOLD Systems database, 99.78% were able to be identified at a specific level, demonstrating a high success rate for egg identification. The samples resulted in 25 species, 11 families, and 2 orders. Of the 25 species found, more than half (60%) present reproductive migration behavior, indicating that the tributaries of the Capivara reservoir are being used as a migratory route by these species. Eggs of rare and endangered species were found, indicating these tributaries as spawning grounds for these species. The results demonstrate the importance of identifying fish eggs in reservoir-influenced environments to recognize breeding areas of native and endangered species, as well as the importance of the Tibagi and Cinzas Rivers for the maintenance of native fish species in the Paranapanema River.
Many tumor cells express chemokines and chemokine receptors, and these molecules can contribute to distinct modes of metastasis processes. It is known that they play a crucial role in breast cancer ...(BC) tumorigenesis and progression. Considering this, it was investigated a possible role for C–Chemokine receptor type 5(
CCR5
) polymorphism
(rs333/delta32)
by conventional polymerase chain reaction (PCR) and CCL5 (C–C motif chemokine ligand 5) protein level by immunosorbent assay (ELISA) in 47 BC patients (resulting in 47 tumoral tissue samples and 47 adjacent normal tissue samples). There was a significant difference between CCL5 level in tumoral and adjacent normal tissues for the same BC patients (p < 0.0001). A significant association was also found for CCL5 level in relation to lymph nodes commitment (p = 0.03). Likewise, there was a significant difference in CCL5 level from tumor tissue of stage III in relation to stage I (p < 0.02). On the other hand, it was verified that
CCR5
-
delta32
polymorphism presented no significant association in relation to CCL5 protein level. Considering the present findings, we suggest that CCL5 may be involved in BC staging and metastasis process.
CXCL12/CXCR4 signaling has been implicated in breast carcinogenesis, and genetic polymorphisms in these molecules have been associated with different types of cancer. The present study analyzed ...genetic polymorphisms in
CXCL12
(rs1801157, G > A) and
CXCR4
(rs2228014, C > T) and CXCR4 immunostaining in tumor tissues from patients with triple negative breast cancer (TNBC) aiming to evaluate their possible role in its’ susceptibility and prognosis. Genetic polymorphisms were analyzed in 59 TNBC patients and 150 control women; age-adjusted logistic regression showed no association when variants were considered in isolation; however, a statistically significant interaction was noted for heterozygosis for both allelic variants increasing the odds for TNBC (CXCL12-GA by CXCR4-CT: OR 7.23; 95% CI 1.15–45.41;
p
= 0.035). CXCL12 polymorphism was correlated negatively with proliferation index (Ki67) (Tau-b = − 0.406;
p
= 0.006). CXCR4 immunostaining was evaluated in 37 TNBC patients (22 with paired tumor-normal adjacent tissue). CXCR4 was detected more intensely in cell cytoplasm than in membrane, and was more expressed in tumor than in normal adjacent tissues, although not statistically significant. CXCR4 expression on the membrane of tumor cells was correlated positively with histopathological grade (Tau-b = 0.271;
p
= 0.036) and negatively with lymph node metastasis (Tau-b = − 0.478;
p
= 0.036). The present study indicates that
CXCL12
and
CXCR4
polymorphisms and CXCR4 immunostaining might have susceptibility and prognostic roles in TNBC pathogenesis.
Background
The HER2 (human epidermal growth factor receptor‐2) Ile655Val (rs1136201) genetic polymorphism can alter the receptor structure and its auto‐activation, which can modify the signal ...transduction and, consequently, the cell cycle regulation. For this reason, this polymorphism has been extensively investigated as a candidate marker for breast cancer (BC). In this context, the aim of this study was to evaluate the possible influence of HER2 Ile655Val in BC susceptibility and prognostic factors in a Brazilian population.
Methods
Polymorphism genotype was assessed through RFLP‐PCR in 107 BC patients with clinicopathological data available and in 150 women with no evidence of neoplasia and with no familial history of BC as control group. Association between this polymorphism and BC susceptibility and clinical parameters was evaluated through odds ratio (OR) and chi‐squared or Fisher's exact test, respectively.
Results
A significant negative association between valine allele and BC susceptibility in dominant model was found (OR 0.5; 95% CI 0.27‐0.93, P = .036). No significant association was found in relation to BC clinicopathological features (tumor size, lymph nodes commitment, histological grade, HER2 overexpression, hormonal receptors, p53, and Ki‐67).
Conclusion
Although this polymorphism did not demonstrate potential as a prognostic marker, it may be a suitable susceptibility marker for BC.
Background
The
HER
2
(human epidermal growth factor receptor‐2) Ile655Val (rs1136201) genetic polymorphism can alter the receptor structure and its auto‐activation, which can modify the signal ...transduction and, consequently, the cell cycle regulation. For this reason, this polymorphism has been extensively investigated as a candidate marker for breast cancer (
BC
). In this context, the aim of this study was to evaluate the possible influence of
HER
2
Ile655Val in
BC
susceptibility and prognostic factors in a Brazilian population.
Methods
Polymorphism genotype was assessed through
RFLP
‐
PCR
in 107
BC
patients with clinicopathological data available and in 150 women with no evidence of neoplasia and with no familial history of
BC
as control group. Association between this polymorphism and
BC
susceptibility and clinical parameters was evaluated through odds ratio (
OR
) and chi‐squared or Fisher's exact test, respectively.
Results
A significant negative association between valine allele and
BC
susceptibility in dominant model was found (
OR
0.5; 95%
CI
0.27‐0.93,
P
= .036). No significant association was found in relation to
BC
clinicopathological features (tumor size, lymph nodes commitment, histological grade,
HER
2 overexpression, hormonal receptors, p53, and Ki‐67).
Conclusion
Although this polymorphism did not demonstrate potential as a prognostic marker, it may be a suitable susceptibility marker for
BC
.
In the present study, a microsatellite-enriched genomic library was constructed and primers for 14 microsatellite loci were designed for
Xylocopa
frontalis
. Twenty unrelated individuals were ...screened. All loci were polymorphic and the number of alleles per locus ranged from 6 to 17 (x = 10.43). Observed (H
o
) and expected (H
e
) heterozygosities ranged from 0.350 to 0.950 and 0.674 to 0.898, respectively. All loci were in Hardy–Weinberg equilibrium, except one. The microsatellite loci described in this study will contribute towards general biology studies of
X. frontalis
, intranidal genetic relationships and nest management for the pollination of passion fruit.
Diversos estudos conservacionistas têm utilizado marcadores moleculares na avaliação da diversidade e estrutura genética de populações de vários organismos. Entretanto, em relação às abelhas da ...subtribo Euglossina, importantes polinizadores neotropicais, ainda são poucos os estudos empregando tais marcadores para um maior conhecimento da diversidade genética de populações destas abelhas. O
presente trabalho investigou a diversidade genética de Euglossa fimbriata de seis fragmentos florestais do norte do estado do Paraná, por meio de marcadores moleculares RAPD e marcadores PCR-RFLP, estes para a análise da região 16S do mtDNA, desta espécie euglossina. Foram utilizados 123 machos de E. fimbriata pertencentes a seis fragmentos florestais constituindo remanescentes de Mata Atlântica, no norte do Paraná, sul do Brasil. As estimativas de proporção de locos polimórficos das seis amostras de E. fimbriata estudadas variaram de 34,44% a 47,78%, enquanto que os valores de heterozigosidade média encontrados variaram de 0,0936 a 0,1527. Com base na análise da AMOVA envolvendo o conjunto de abelhas dos seis fragmentos florestais, detectou-se que a quantidade de variação genética foi maior dentro dos grupos (95,41%) do que entre os grupos de abelhas amostrados (4,59%) Foram utilizadas duas enzimas de restrição (AseI e DraI) para o corte da região 16S do mtDNA. Foram encontrados 4 haplótipos gerados pela enzima AseI e 2 haplótipos pela enzima DraI. Combinados estes haplótipos constituíram 5 haplótipos-compostos presentes de forma variada entre os grupos de abelhas dos diferentes fragmentos de mata estudados. Um dos haplótiposcompostos encontrados mostrou-se presente entre as amostras de abelhas de todos
os fragmentos florestais estudados, ocorrendo em freqüências entre 63,64 a 95,65%. A ampla distribuição e elevada freqüência de tal haplótipo sugere sua origem ancestral em comparação aos demais haplótipos-compostos encontrados. Os resultados obtidos com ambos os tipos de marcadores moleculares (RAPD e PCR-RFLP) indicam a existência de subpopulações constituindo uma metapopulação de E. fimbriata na região estudada, bem como a importância de se preservar o conjunto de fragmentos florestais (independente do tamanho e grau de
interferência antrópica destes) para a manutenção da diversidade genética desta espécie de Euglossina no norte do Paraná.
Molecular markers are widely used in conservation studies to assess the genetic diversity and population structure of different organisms. However, studies using these markers to assess the genetic diversity of euglossine bees, which are important neotropical pollinators, are still scarce. Thus, the aim of the current study was to evaluate the genetic variation and genetic structure of Euglossa fimbriata, an euglossine bee species which shows a wide distribution throughout Brazilian forest remnants, by mean of RAPD and PCR-RFLP markers. A total of 123 males of E. fimbriata from six fragments of Atlantic Forest, located at Northern Paraná State, Southern Brazil, were used in the analysis. The proportion of polymorphic loci in each sample, assayed by RAPD markers, varied from 34.44% to 47.78%, while the
estimates of average heterozygosity ranged from 0.0936 to 0.1527. AMOVA analysis revealed that genetic variation was higher within samples (95.41%) than among samples (4.59%). Two restriction enzymes (AseI e DraI) were used for the digestion of 16S mtDNA region. Respectively, 4 and 2 haplotypes were detected by endonucleases AseI and DraI. The association of both restriction enzymes resulted
in 5 composite-haplotypes distributed irregularly among the groups of bees from different fragments. One composite-haplotype was found among bees from all forest remnants, occurring in frequencies that varied from 63.64 to 95.65%. These patherns of variation suggest an older origin of this haplotype when compared to others composite-haplotypes found. The results obtained with RAPD and PCR-RFLP
markers suggest the occurrence of subpopulations of E. fimbriata, constituting a metapopulation structure of this euglossine species throughout the region studied and reinforce the need of better protecting these forest remnants (regardless of their reduced size and degree of anthropogenic interference) to maintain the genetic diversity of this euglossine species in Northern Paraná State.
Euglossa constitui o gênero mais diverso em número de espécies da tribo Euglossini, com mais de 110 espécies reconhecidas. Porém, algumas questões têm sido levantadas em relação às espécies de ...Euglossa. O presente trabalho fez uso de diferentes técnicas moleculares para tentar responder algumas questões genéticas envolvendo as abelhas Euglossini, em destaque: questões taxonômicas e filogenéticas, especialmente as relacionadas a várias espécies de Euglossa com ocorrência reconhecida para a Mata Atlântica; e, aspectos ligados à origem de indivíduos ginandromorfos e intersexos. O presente trabalho utilizou o sequenciamento de parte das regiões COI, CytB e 16S rDNA do DNA mitocondrial e região 28S rDNA do DNA nuclear a fim de tentar elucidar o real status taxonômico de Euglossa iopoecila e Euglossa roubiki. Foi realizada a análise de distância genética segundo o modelo K2P, para a região barcode de Euglossa iopoecila e Euglossa roubiki bem como análises filogenéticas (máxima parcimônia (MP), máxima verossimilhança (ML) e análise bayesiana). Os dados da região barcode, do gene CytB e dos genes concatenados foram utilizados para a construção de redes de haplótipos. Os dados concatenados de todos os genes resultaram em 3125 caracteres utilizados nas análises. Os valores baixos (<0,3%) de distância genética (modelo K2P) encontrados entre os indivíduos de E iopoecila e E. roubiki e as análises de máxima parcimônia, máxima verossimilhança e bayesiana, bem como as redes de haplótipos indicam ser estas uma mesma espécie. Igualmente, foi utilizado o sequenciamento de parte das regiões COI, 16S e 28S, para delinear a filogenia de espécies do gênero Euglossa da Mata Atlântica e, realizar as análises filogenéticas (MP, ML e análise bayesiana). Os dados concatenados de todos os genes resultaram em 2311 caracteres utilizados nas análises. As árvores filogenéticas encontradas no presente trabalho sustentam os resultados de outros que apontam Euglossella como grupo irmão dos outros subgêneros de Euglossa; ainda, alguns ramos se mostraram parafiléticos corroborando os resultados de outros trabalhos, que apontam um agrupamento entre Glossura, Glossurella e Glossuropoda. Os resultados obtidos reforçam a necessidade de uma revisão dos subgêneros de Euglossa. Para o estudo de um indivíduo ginandromorfo de Euglossa melanotricha foram utilizados nove marcadores microssatélites, que indicaram ser este um organismo haploide ou homozigoto para os locos analisados. Em relação ao indivíduo de E. melanotricha, os resultados indicam se tratar de um organismo intersexo, pois este possui partes femininas e masculinas (tecidos) e é geneticamente uniforme.
Euglossa is the most diverse genus in number of species of the tribe Euglossini, with more than 110 species described. However, some questions have been raised regarding Euglossa species. Therefore, this study describes different molecular techniques adressed to answer some genetic questions about Euglossini bees, including: taxonomic and phylogenetic aspects involving different Euglossa species with recognized occurrence to Atlantic Forest domain; and, aspects potentially related to the genetic of gynandromorph and intersex individuals. We used sequencing of regions of COI and 16S rDNA CytB mitochondrial DNA and 28S rDNA regions of nuclear DNA to elucidate the taxonomic status of Euglossa iopoecila and Euglossa. roubiki. We performed the analysis of genetic distance according to K2P model for the barcode region of E. iopoecila and E. roubiki and phylogenetic analyzes (maximum parsimony (MP), maximum likelihood (ML) and Bayesian analysis). Data from the barcode, CytB gene and genes concatenated were used to construct haplotype networks. The concatenated data of all genes resulted in 3,125 characters used in the analysis. Low values (<0.3%) of genetic distance were identified between individuals of E. iopoecila and E. roubiki while MP, ML, Bayesian analyses and haplotype networks suggest that they belong to the same species.Also, we used the sequencing of the regions COI, 16S and 28S to perform phylogenetic analysis (MP, ML and Bayesian analysis) of Euglossa, including several species from Atlantic Forest biome. The concatenated data of all genes resulted in 2,311 characters used in the analysis. The phylogenetic analyses support the results of other studies that consider Euglossella as sister group of the other subgenera of Euglossa; also the identification of some paraphyletic branches corroborate the results of other studies, which indicate a group between Glossura, Glossurella and Glossuropoda. The results suggest the need for a more detailed revision of the subgenera of Euglossa. Nine microsatellites were used to analyze an anomalous individual of E. melanotricha, which revealed to be a homozygous or a hemizygous for all loci analyzed. Our findings indicate that the organism of E. melanotricha analyzed is an intersex, since both female and male body parts (tissue) of this individual were genetically uniform.
CCN2 is best known as a promoter of chondrocyte differentiation among the CCN family members, and Ccn2 null mutant mice display skeletal dysmorphisms. However, little is known concerning the roles of ...CCN2 during bone formation. We herein present a comparative analysis of wild-type and Ccn2 null mice to investigate the roles of CCN2 in bone development. Multiple histochemical methods were employed to analyze the effects of CCN2 deletion in vivo, and effects of CCN2 on the osteogenic response were evaluated with the isolated and cultured osteoblasts. As a result, we found a drastic reduction of the osteoblastic phenotype in Ccn2 null mutants. Importantly, addition of exogenous CCN2 promoted every step of osteoblast differentiation and rescued the attenuated activities of the Ccn2 null osteoblasts. These results suggest that CCN2 is required not only for the regulation of cartilage and subsequent events, but also for the normal intramembranous bone development.
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