Polymorphism of the Prolactin (PRL) Gene and Its Effect on Milk Production Traits in Romanian Cattle Breeds

In the present study, we investigated one polymorphism of the PRL gene (rs211032652 SNP) and assessed its influence on milk production and chemical composition in two Romanian cattle breeds. A total of 119 cattle from two breeds reared in Western Romania (64 Romanian Spotted and 55 Romanian Brown) were included in the research herd. A PCR-RFLP genotyping assay was used for the identification of the rs211032652 SNP variants. Shapiro’s test and Levene’s test were used to verify ANOVA assumptions and ANOVA and Tukey’s test were employed to test the associations between PRL genotypes and five milk traits. Among the studied breeds, our results showed that PRL genotypes were significantly associated (p < 0.05) with fat and protein percentage in the milk of Romanian Brown cattle. The AA genotype was associated with a higher fat percentage in milk (4.76 ± 0.28) compared to the GG genotype (4.04 ± 0.22, p = 0.048), as well as a higher protein percentage (3.96 ± 0.32% vs. 3.43 ± 0.15%, p = 0.027) in Romanian Brown cattle. Moreover, the PRL locus favored a significantly higher fat (p = 0.021) and protein (p = 0.028) percentage in the milk of Romanian Brown cattle compared to the Romanian Spotted breed, with a difference of 0.263% and 0.170%, respectively

DETERMINATION OF PLASMA PHENYLALANINE CONCENTRATION BY TWO DIMENSIONALTHIN LAYER CHROMATOGRAPHY AND HIGH PERFORMANCE LIQUID CHROMATOGRAPHY IN RELATION WITH DIAGNOSIS OF PHENYLKETONURIA

Phenylketonuria (PKU) is a genetic disorder which can lead to serious long-term complications in children, including intellectual impairment. The cerebral damage can be largely eliminated if hyperphenilalaninemia (HPA) and PKU is detected by newborn screening and the dietary treatment (started no later than 10 days after birth), brings within 15 days the blood Phe level in the ranges 2 - 6 mg/dL (or equivalently 120 - 360 µmol/L). Consequently, the measurement of plasma Phe level in children with PKU is of utmost importance. We present here in detail two methods for determination of plasma Phe concentrations in children with HPA (detected by newborn screening) or suspected to have PKU based on clinical symptoms. The first method, involving two-dimensional thin-layer chromatography (2D - TLC) on micro scale chromatograms, allows the identification of patients with PKU. However, the quantitation of plasma Phe concentration by the 2D - TLC video-densitometric method is only semiquantitative, because is accompanied by large errors. In contrast, the high performance liquid chromatography determination of the plasma Phe level is fast, very sensitive and higly accurate. Both methods are rather cheap and reliable, suitable for use in countries that cannot afford more expensive procedures for diagnosis and monitoring of PKU

Microbial cellulases immobilized in biopolymer/silica matrices used as enzyme release systems

Trichoderma viride CMGB 1 cellulases were immobilized by entrapment in silica gels (by sol-gel method), alginate biopolymers and hybrid alginate/silica materials. Tetramethoxysilane (TMOS), tetraethoxysilane (TEOS) and tetrakis (2-hydroxyethyl) orthosilicate (THEOS) were used as organoalkoxysilane precursors and ethanol or ethylene glycol as cosolvents in a two step sol-gel synthesis. Combined alginate/silica matrices resulted by mixing silica sol with sodium alginate or by coating alginate beads with a silica shell. The partial confinement of ethylene glycol in the matrix with consequences on biocatalytic activity was investigated using SEM-EDAX, thermal analysis and FT-IR spectroscopy. The efficiency of the enzyme-matrix biomaterials was tested in controlled enzyme release experiments.&#x0D; The sol-gel method developed using EG as a co-solvent allowed cellulase immobilization yields 1.5-4.5 times higher compared to classical sol-gel methods that use EtOH. The characterization of the gels by microscopic and spectrophotometric analyzes showed that there are similarities between the structure of the gels based on THEOS and those developed by us from TEOS, TMOS and EG as co-solvent. The new developed gels showed good cellulase release properties at acidic pH, comparable to those based on THEOS and alginate. The microbial cellulases immobilized in the matrices obtained and characterized in this work can operate as efficient systems for releasing enzymes, in acidic pH conditions, as feed additives.</jats:p

Effects of Simulated Gastric Acid Exposure on Surface Topography, Mechanical and Optical Features of Commercial CAD/CAM Ceramic Blocks

Gastric acid exposure produces tooth structure demineralization and dental ceramic degradation. The most affected patients are those who suffer from gastroesophageal reflux disease, bulimia nervosa, and pregnant women with prolonged severe nausea. In order to protect this kind of patient, the purpose of this study was to determine whether simulated gastric acid exposure leads to microstructural changes in surface topography, hardness, color changes, and translucency of some ceramic materials, which are already on the market. Forty disks (Triluxe Forte, Cerasmart, Enamic and Empress CAD) were analyzed before and after immersion in simulated gastric acid juice, in terms of microhardness, surface roughness, translucency, and surface morphology using scanning electron microscopy. Color change was assessed by using a spectrophotometer based on CIELab parameters and the results showed that, after exposure, ∆Eab remained under the threshold of acceptability and perceptibility. In terms of microhardness, Cerasmart is the only material that did not undergo changes after immersion. SEM images illustrated observable changes surface topography after acid exposure for all the tested materials. In conclusion, Triluxe Forte suffered the most important changes after simulated gastric acid juice exposure, and Cerasmart proved to be the least affected material.</jats:p

Effects of Simulated Gastric Acid Exposure on Surface Topography, Mechanical and Optical Features of Commercial CAD/CAM Ceramic Blocks

Gastric acid exposure produces tooth structure demineralization and dental ceramic degradation. The most affected patients are those who suffer from gastroesophageal reflux disease, bulimia nervosa, and pregnant women with prolonged severe nausea. In order to protect this kind of patient, the purpose of this study was to determine whether simulated gastric acid exposure leads to microstructural changes in surface topography, hardness, color changes, and translucency of some ceramic materials, which are already on the market. Forty disks (Triluxe Forte, Cerasmart, Enamic and Empress CAD) were analyzed before and after immersion in simulated gastric acid juice, in terms of microhardness, surface roughness, translucency, and surface morphology using scanning electron microscopy. Color change was assessed by using a spectrophotometer based on CIELab parameters and the results showed that, after exposure, ∆Eab remained under the threshold of acceptability and perceptibility. In terms of microhardness, Cerasmart is the only material that did not undergo changes after immersion. SEM images illustrated observable changes surface topography after acid exposure for all the tested materials. In conclusion, Triluxe Forte suffered the most important changes after simulated gastric acid juice exposure, and Cerasmart proved to be the least affected material

Comparative Analysis of Romanian and Swiss Bovine Populations Using Whole Genome Sequencing and SNP Microarrays

The use of SNP microarrays has gained distinguished attention in recent years and the identification of numerous SNPs has proven valuable for genetic evaluation and selection in farm animals. In the current study we compared several bovine populations from Romania and Switzerland at the level of SNPs. Romanian Brown (N=39) and Romanian Spotted (N=245) cattle were genotyped using the Axiom Bovine BovMDv3 SNP microarray. For the Swiss population, we acquired sequencing data from the NCBI SRA database for 80 individuals from three breeds: Brown Swiss (N=20), Original Braunvieh (N=20) and Simmental (N=40). Sequencing data were processed using the Bcbio-nextgen data analysis pipeline and variants were called based on the UMD3.1 reference genome. Common SNPs found from both microarray and sequencing data were retained and genotypes from all the Romanian and Swiss animals were pooled together, resulting in a combined dataset of 48,291 SNPs for 364 individuals. Pairwise comparisons were assessed on the five subpopulations according to Weir and Cockerham’s FST index. Small genetic differences (FST < 0.05) were found between the Romanian Brown and Swiss Brown Swiss subpopulations and between Romanian Spotted and Swiss Simmental subpopulations. For all the other pairwise comparisons, FST values were between 0.05 and 0.1, indicating a moderate level of genetic difference among the corresponding subpopulations. The results of fastSTRUCTURE indicated that the most likely number (K) of subpopulations from the pooled dataset was between 8 and 12. Bar plots for K = 5, 8 and 12 confirmed that Romanian Brown and Swiss Brown Swiss subpopulations were genetically similar. However, they also revealed a surprisingly high level of heterogeneity among the Romanian Spotted individuals. As such, future research is required to zoom in on the genetic make-up and explain the most likely sources of heterogeneity for the Romanian Spotted breed. Current results will facilitate a better understanding of genomic selection and its application for improved breeding programs in Romanian cattle breeds