PCR-SSCP analysis of GH gene in Sarda goats: a high variability and its preliminary effects on dairy performances

The growth hormone (GH) gene can be utilized as a major gene because in various domestic livestock its polymorphisms have been associated to milk traits. The aim of this research was to investigate single-strand conformation polymorphism (SSCP) in the exon 3 of gGH (goat GH) gene and to evaluate the possible association with milk traits in Sarda goat breed. Forty-four primiparous lactating goats were randomly chosen, and the productive parameters (milk yield, fat, protein, and lactose percentage) of three consecutive lactations were monitored. The exon 3 of the gGH gene was PCR amplified and the resulting products were analysed by SSCP. Six conformational patterns were detected. The sequencing of SSCP patterns revealed the occurrence of six nucleotide changes, two of which determined amino acid changes in the deduced protein sequence. A preliminary comparative analysis of the productive traits related to three lactations with the genomic profiles derived from the SSCP analysis was performed with the ANOVA statistical method. SSCP polymorphic patterns in exon 3 were associated (P<0.01) with milk yield, fat and protein percentages, and with lactose content (P<0.05). These findings may be used for marker assisted selection in Sarda goat, in order to improve dairy production, preserving genetic diversity of the population

Reproductive activity in sheep with different lambing period treated with melatonin in April

The object was to evaluate the effect of melatonin treatment on the advance in April of the reproductive resumption in Sarda breed sheep with different lambing period. For the research two farms, located in North Sardinia between 39° and 40° N, were chosen. In each farm, 120 lactating ewes were selected: 30 lambed between October 20th and November 20th (group 1); 30 lambed between December 1st and 30th (group 2); 30 lambed between January 1st and 30th; 30 lambed between February 1st and 28th (group 4). In each farm, each group of 30 animals was divided into two subgroups of 15 animals (M and C). On April 1st, in each farm, the animals of the M subgroups were treated with a implant containing 18 mg melatonin. The subgroups C were kept as control. The lambing dates and the number of newborn lambs were recorded until 220 days after ram introduction. In treated animals greatest fertility (P<0.01) and lowest distance in days from male introduction to lambing (P<0.01) were recorded. The best reproductive performances were found in the group 1 and 2 compared to the other two groups (P<0.01).The present research shows that melatonin treatment should be made 3 or 4 months after lambing, in order to obtain optimal results

Polymorphism of Caprine SLC11A1 Gene and Relationships with Hygienic Characteristics of Milk

The solute carrier family 11 member A1 (SLC11A1) gene is associated with resistance to infectious diseases. Genetic variability at the 3’ untranslated region (3’-UTR) of this gene is due to the presence of a polymorphic microsatellites that contain a (GT) n dinucleotide repeat. The microsatellite variability and relationships with milk yield and composition, somatic cell count (SCC) and total microbic count (TMC) were investigated in 260 goats of Sarda breed. Genotyping of the upstream guanine-thymine repeat (GT)n revealed twenty different genotypes and eight alleles (GT11, GT12, GT14, GT15, GT16, GT17, GT18 and GT19). The present study confirmed the high genetic variability of the Sarda goat and that the genotype of the microsatellite at 3’-UTR SLC11A1 affected many chemical and hygienic characteristics of milk as fat, protein and SCC

Polymorphism of Caprine SLC11A1 Gene and Relationships with Hygienic Characteristics of Milk

The solute carrier family 11 member A1 (SLC11A1) gene is associated with resistance to infectious diseases. Genetic variability at the 3’ untranslated region (3’-UTR) of this gene is due to the presence of a polymorphic microsatellites that contain a (GT) n dinucleotide repeat. The microsatellite variability and relationships with milk yield and composition, somatic cell count (SCC) and total microbic count (TMC) were investigated in 260 goats of Sarda breed. Genotyping of the upstream guanine-thymine repeat (GT)n revealed twenty different genotypes and eight alleles (GT11, GT12, GT14, GT15, GT16, GT17, GT18 and GT19). The present study confirmed the high genetic variability of the Sarda goat and that the genotype of the microsatellite at 3’-UTR SLC11A1 affected many chemical and hygienic characteristics of milk as fat, protein and SCC

Tumor suppressor genes are frequently methylated in lymph node metastases of breast cancers

<p>Abstract</p> <p>Introduction</p> <p>Metastasis represents a major adverse step in the progression of breast carcinoma. Lymph node invasion is the most relevant prognostic factor; however little is known on the molecular events associated with lymph node metastasis process. This study is to investigate the status and role of methylation in lymph node metastatic tumors.</p> <p>Materials and methods</p> <p>Bisulfite pyrosequencing is used to screen 6 putative tumor suppressor genes (<it>HIN-1, RASSF1A, RIL, CDH13, RARβ2 </it>and E-cadherin) in 38 pairs of primary breast tumors and lymph node metastases.</p> <p>Results</p> <p>We found that <it>HIN-1, CDH13, RIL, RASSF1A </it>and <it>RARβ2 </it>were frequently methylated both in primary and metastatic tissues (range: 55.3%~89.5%). E-cadherin was not frequently methylated in either setting (range: 18.4%~23.7%). The methylation status of <it>HIN-1, CDH13, RIL</it>, and <it>RARβ2 </it>in lymph nodes metastasis were correlated with that in primary tumors. The Pearson correlation values ranged from 0.624 to 0.472 (<it>p </it>values < 0.01 to 0.001). Interestingly, we observed an association between <it>HIN-1 </it>methylation and hormone status in metastatic lymph nodes. Hypermethylation of <it>HIN-1 </it>in metastasis lymph nodes was significantly associated with expression of ER (odds ratio, 1.070; P = 0.024) and with PR (odds ratio, 1.046; P = 0.026).</p> <p>Conclusions</p> <p>This study suggests that hypermethylation of tumor suppressor genes is extended from primary to metastatic tumors during tumor progression.</p

Genomic signatures for paclitaxel and gemcitabine resistance in breast cancer derived by machine learning.

Increasingly, the effectiveness of adjuvant chemotherapy agents for breast cancer has been related to changes in the genomic profile of tumors. We investigated correspondence between growth inhibitory concentrations of paclitaxel and gemcitabine (GI50) and gene copy number, mutation, and expression first in breast cancer cell lines and then in patients. Genes encoding direct targets of these drugs, metabolizing enzymes, transporters, and those previously associated with chemoresistance to paclitaxel (n = 31 genes) or gemcitabine (n = 18) were analyzed. A multi-factorial, principal component analysis (MFA) indicated expression was the strongest indicator of sensitivity for paclitaxel, and copy number and expression were informative for gemcitabine. The factors were combined using support vector machines (SVM). Expression of 15 genes (ABCC10, BCL2, BCL2L1, BIRC5, BMF, FGF2, FN1, MAP4, MAPT, NFKB2, SLCO1B3, TLR6, TMEM243, TWIST1, and CSAG2) predicted cell line sensitivity to paclitaxel with 82% accuracy. Copy number profiles of 3 genes (ABCC10, NT5C, TYMS) together with expression of 7 genes (ABCB1, ABCC10, CMPK1, DCTD, NME1, RRM1, RRM2B), predicted gemcitabine response with 85% accuracy. Expression and copy number studies of two independent sets of patients with known responses were then analyzed with these models. These included tumor blocks from 21 patients that were treated with both paclitaxel and gemcitabine, and 319 patients on paclitaxel and anthracycline therapy. A new paclitaxel SVM was derived from an 11-gene subset since data for 4 of the original genes was unavailable. The accuracy of this SVM was similar in cell lines and tumor blocks (70-71%). The gemcitabine SVM exhibited 62% prediction accuracy for the tumor blocks due to the presence of samples with poor nucleic acid integrity. Nevertheless, the paclitaxel SVM predicted sensitivity in 84% of patients with no or minimal residual disease

Characterization of the SREBP-1 Gene Polymorphisms and Milk Traits in Dairy Sheep

The SREBP genes (Sterol Regulatory Element-Binding Proteins) are involved in the milk fat synthesis. In dairy cows some polymorphisms at the SREBP-1 gene sequence have been related with milk fat content. The aim of this study was to characterize the entire coding regions of the SREBP-1 gene in Sarda sheep breed, in order to highlight any polymorphisms and their association with milk traits. Four-hundred adult and lactating Sarda ewes were selected. Individual milk yield was recorded monthly from Day 30 to Day 150 of lactation, and fat and protein concentration were analysed. A blood sample from each ewe was taken for DNA extraction; thus, all the 19 coding exons of the SREBP-1 gene were amplified by polymerase chain reaction (PCR). Single-strand conformation polymorphism analysis (SSCP) and sequencing were used to scan mutations. Results provide, for the first time, the entire coding DNA sequence (CDS) of the SREBP-1 gene in sheep, and by sequences analysis 8 polymorphisms have been detected. The statistical analysis exhibited no relationship between polymorphisms and milk traits. The low SREBP-1 gene diversity that emerged from the present study, may be linked to the important role of this gene in the mechanism of milk fat synthesis or to the severe genetic selection performed in the Sarda sheep. However, it would be necessary to extend the study, including other breeds and other genes, in order to expanding the knowledge about the process of milk fat synthesis in dairy sheep

Different biological and prognostic breast cancer populations identified by FDG-PET in sentinel node-positive patients: Results and clinical implications after eight-years follow-up

Abstract Background Sentinel node (SN) biopsy is the standard method to evaluate axillary node involvement in breast cancer (BC). Positron emission tomography with 2-(fluorine-18)-fluoro-2-deoxy-D-glucose (FDG-PET) provides a non-invasive tool to evaluate regional nodes in BC in a metabolic-dependent, biomolecular-related way. In 1999, we initiated a prospective non-randomized study to compare these two methods and to test the hypothesis that FDG-PET results reflect biomolecular characteristics of the primary tumor, thereby yielding valuable prognostic information. Patients and methods A total of 145 cT1N0 BC patients, aged 24–70 years, underwent FDG-PET and lymphoscintigraphy before surgery. SN biopsy was followed in all cases by complete axillary dissection. Pathologic evaluation in tissue sections for involvement of the SN and other non-SN nodes served as the basis of the comparison between FDG-PET imaging and SN biopsy. Results FDG-PET and SN biopsy sensitivity was 72.6% and 88.7%, respectively, and negative predictive values were 80.5% and 92.2%, respectively. A subgroup of more aggressive tumors (ER-GIII, Her2+) was found mainly in the FDG-PET true-positive (FDG-PET+) patients, whereas LuminalA, Mib1 low-rate BCs were significantly undetected ( p = 0.009) in FDG-PET false-negative (FDG-PET−) patients. Kaplan–Meier survival estimates after a median follow-up of more than 8 years showed significantly worse overall survival for FDG-PET+ patients in node-positive (N+) patients ( p = 0.035) as compared to N+/FDG-PET− patients, which overlapped with survival curves of N− and FDG-PET+ or − patients. Conclusions Our findings suggest that FDG-PET results reflect intrinsic biologic features of primary BC tumors and have prognostic value with respect to nodal metastases. FDG-PET false negative cases appear to identify less aggressive indolent metastases. The possibility to identify a subgroup of N+ BC patients with an outcome comparable with N− BC patients could reduce the surgical and adjuvant therapeutic intervention

Whole-transcriptome analysis links trastuzumab sensitivity of breast tumors to both HER2 dependence and immune cell infiltration.

While results thus far demonstrate the clinical benefit of trastuzumab, some patients do not respond to this therapy. To identify a molecular predictor of trastuzumab benefit, we conducted whole-transcriptome analysis of primary HER2+ breast carcinomas obtained from patients treated with trastuzumab-containing therapies and correlated the molecular portrait with treatment benefit. The estimated association between gene expression and relapse-free survival allowed development of a trastuzumab risk model (TRAR), with ERBB2 and ESR1 expression as core elements, able to identify patients with high and low risk of relapse. Application of the TRAR model to 24 HER2+ core biopsies from patients treated with neo-adjuvant trastuzumab indicated that it is predictive of trastuzumab response. Examination of TRAR in available whole-transcriptome datasets indicated that this model stratifies patients according to response to trastuzumab-based neo-adjuvant treatment but not to chemotherapy alone. Pathway analysis revealed that TRAR-low tumors expressed genes of the immune response, with higher numbers of CD8-positive cells detected immunohistochemically compared to TRAR-high tumors. The TRAR model identifies tumors that benefit from trastuzumab-based treatment as those most enriched in CD8-positive immune infiltrating cells and with high ERBB2 and low ESR1 mRNA levels, indicating the requirement for both features in achieving trastuzumab response