Oxidative Status of Goats with Different CSN1S1 Genotypes Fed ad Libitum with Fresh and Dry Forages

Forty late-lactation Girgentana goats were used to study the effect of diets fed ad libitum and αS1-casein (CSN1S1) genotype on redox balance. The goats genotyped at CSN1S1 locus (A/A, A/F) were subjected to four feeding treatments different for percentage inclusion of dry and fresh forage: DAF100 (98% of Dry Alfalfa Forage), DAF65 (65% of Dry Alfalfa Forage), FSF100 (100% of Fresh Sulla Forage) and FSF65 (65% of Fresh Sulla Forage). Blood samples were analyzed for superoxide dismutase (SOD) and glutathione peroxidase (GPX) activity, reactive oxygen metabolites (ROMs), biological antioxidant potential (BAP) and non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHBA), albumin, glucose and cholesterol contents. The oxidative stress index (OSI) was calculated as percentage ratio of ROMs to BAP. Redox balance was improved by Sulla inclusion, as reflected in the lower OSI values found in FSF100 and FSF65 groups. DAF100 group displayed the highest GPX activity, while other groups exhibited the highest SOD activity. Fresh forage diets increased albumin concentration while no effect of tested factors was noted on glucose, NEFA, BHBA and cholesterol contents. The interaction diet × genotype was significant only for GPX activity. GPX and albumin were negatively correlated and were correlated positively and negatively with ROMs, respectively. Diet rather than genotype affects redox balance in dairy goats and a possible role of forage polyphenol compounds on oxidative status needs to be tested in future studies

Effect of diet and CSN1S1 genotype on nutritional, productive and metabolic responses of milking Girgentana goats.

From a group of milking goats genotyped using specific PRC protocols at DNA level, 12 goats, averaging 37.2±3.5 kg of live weight, were selected for having the same genotype at CSN1S2, CSN2 and CSN3 loci and differing for the CSN1S1 genotype (G): 6 goats were homozygous for a strong allele (AA) and 6 heterozygous for a weak alleles (AF). Goats of each genotype were allocated homogeneously, based on days in milking (DIM, 50 or 120 days), to 3 sub-groups and fed ad libitum in individual pen with 3 diets, in a 3 x 3 Latin square design with 3 periods (P) comprised of 14 days for adaptation and 7 days for data and samples collection. The diets (D) were sulla (Hedysarum coronarium L.) fresh forage (SFF), sulla fresh forage plus 800 g/d of barley meal (SFB), mixed hay plus 800 g/d of barley meal (MHB), with 130, 95 and 85 protein/energy ratio (P/E, g protein/Mcal net energy), respectively. During experimental period, milk production and feed intake were measured every day, and milk quality was detected three times. Blood samples were collected at the end of pre-experimental and experimental periods. Plasma content of NEFA, glucose, insulin, fT3 and fT4 was detected; also same plasma markers of oxidative stress were measured as Reactive Oxygen Metabolites (ROMs), biological antioxidant potential (BAP) and -tocopherol. Data were analysed by a mixed model with DIM, P, G, D and GxD as fixed effects, and goat as random effect. Data of pre-experimental period were used as covariates. There was no significant effect of G and interaction GxD on dry matter (DM) and nutrients intake, efficiency of dietary protein utilization for milk casein synthesis (EPU), glucose, NEFA, insulin, ROMs, BAP and tocopherol content. With regard to D effect, DM intake was lower with MHB than SFF and SFB (1655 vs. 1820, 1807 g/d; P<0.01), whereas protein intake increased passing from MHB to SFB and SFF (203 vs. 290 vs. 321 g/d, P<0.01). NE intake was higher in SFB (3.0 vs. 2.3, 2.4 Mcal/d for SFB, MHB, SFF; P<0.01), and ingested NDF was higher in SFF than in MHB and SFB (632 vs. 539, 483 g/d; P<0.01). The diet greatly affected milk yield, which increased from SFF to MHB and SFB (1353 vs. 1423 vs.1664 g/d; P<0.01). The significant effect of GxD on milk yield (P<0.01) was linked to the superiority induced by SFB diet in AA than in AF goats (1720 vs. 1608 g/d; P<0.05). Milk composition was affected by D equally in both genotypes. Barley supplement contributed to reduce fat (36 vs. 32 vs. 30 g/kg in SFF, SFB, MHB; P<0.01) and urea in milk (35 vs, 32, 31 mg/dl in SFF, SFB, MHB; P<0.01), whereas the fresh forage increased the casein content (27, 27 vs. 26 g/kg in SFF, SFB, MHB; P<0.01). The EPU was the highest in MHB group, due to lower protein intake, whereas it was favoured by energy supplement (191 vs. 164 vs. 115 g casein/kg protein intake in MHB, SFB, SFF; P<0.01). The milk of AA goats showed longer coagulation time (r: 15 vs. 14 min; P<0.05) and higher curd firmness (a30: 36 vs. 29 mm; P<0.01) than AF milk. The D affected NEFA, ROMs and BAP. SFF showed the higher NEFA than other diets (0.39 vs. 0.23, 0.21 mmol/l in SFF, MHD, SFB; P<0.01). The BAP increased in groups that utilized fresh forage compared to group fed mixed hay (7.68 vs. 8.01, 7.99 ln mol/l in MHB, SFF, SFB; P<0.05). The ROMs level was lower in SFF goats (3.90 ln U.Carr) compared to MHB and SFB. A significant effect of G (P<0.05) was detected on fT4, which was higher by 15% in the AF than in the AA (1.02 vs. 0.88 ng/dl; P<0.05). A slight increase of fT3 was detected in AA compared to AF (3.58 vs. 3.36 pg/ml; P=0.10). Interaction GxD (P<0.05) was found for fT3, being higher in AA goats than AF goats (3.92 vs. 3.03 pg/ml) but only when fed the MHB diet

Monitoring of Cardiac Output Using a New Smartphone Application (Capstesia) vs. Vigileo FloTrac System

(1) Background: We tested Capstesia against a reference system, Vigileo FloTrac, in patients undergoing major vascular surgery procedures. (2) Methods: Twenty-two adult patients (236 data pairs) were enrolled. Cardiac output (CO), stroke volume (SV), systemic vascular resistance (SVR), and related indexed parameters from the two monitoring systems were collected and compared at eleven time points during surgery. Intraclass correlation coefficients with 95% confidence intervals (CIs) and Bland–Altman plots with percentages of error were used. (3) Results: The interclass correlation coefficients for CO, SV, and SVR were 0.527 (95%CI 0.387 to 0.634), 0.580 (95%CI 0.454 to 0.676), and 0.609 (95%CI 0.495 to 0.698), respectively. In the Bland–Altman analysis, bias (and limits of agreement) of CO was 0.33 L min−1 (−2.44; 3.10), resulting in a percentage error of 61.91% for CO. For SV, it was 5.02 mL (−36.42; 46.45), with 57.19% of error. Finally, the bias (and limit of agreement) of SVR was −75.99 dyne sec cm−5 (−870.04; 718.06), resulting in an error of 69.94%. (4) Conclusions: Although promising, cost-effective, and easy to use, the moderate level of agreement with Vigileo and the high level of error make Capstesia unsuitable for use in the intraoperative setting of vascular surgery. Critical errors in acquisition or digitalization of the snap might have a strong impact on the accuracy and performance. Further standardization of the acquisition technique and improvements in the processing algorithm are needed

Monitoring of Cardiac Output Using a New Smartphone Application (Capstesia) vs. Vigileo FloTrac System

(1) Background: We tested Capstesia against a reference system, Vigileo FloTrac, in patients undergoing major vascular surgery procedures. (2) Methods: Twenty-two adult patients (236 data pairs) were enrolled. Cardiac output (CO), stroke volume (SV), systemic vascular resistance (SVR), and related indexed parameters from the two monitoring systems were collected and compared at eleven time points during surgery. Intraclass correlation coefficients with 95% confidence intervals (CIs) and Bland–Altman plots with percentages of error were used. (3) Results: The interclass correlation coefficients for CO, SV, and SVR were 0.527 (95%CI 0.387 to 0.634), 0.580 (95%CI 0.454 to 0.676), and 0.609 (95%CI 0.495 to 0.698), respectively. In the Bland–Altman analysis, bias (and limits of agreement) of CO was 0.33 L min−1 (−2.44; 3.10), resulting in a percentage error of 61.91% for CO. For SV, it was 5.02 mL (−36.42; 46.45), with 57.19% of error. Finally, the bias (and limit of agreement) of SVR was −75.99 dyne sec cm−5 (−870.04; 718.06), resulting in an error of 69.94%. (4) Conclusions: Although promising, cost-effective, and easy to use, the moderate level of agreement with Vigileo and the high level of error make Capstesia unsuitable for use in the intraoperative setting of vascular surgery. Critical errors in acquisition or digitalization of the snap might have a strong impact on the accuracy and performance. Further standardization of the acquisition technique and improvements in the processing algorithm are needed

Effects of different genotypes at the CSN3 and LGB loci on milk and cheese-making characteristics of the bovine Cinisara breed

The Cinisara is an autochthonous Sicilian bovine breed reared mainly for production of Caciocavallo Palermitano, a typical stretched-curd cheese. The distribution of A and B alleles at both the b-lactoglobulin (LGB) and k-casein (CSN3) loci in Cinisara cows and their influence on milk traits and cheese yield and composition were analysed. The LGB alleles are associated with significantly different effects on whey protein level (lower for BB genotype, P<0.01), and casein index (higher value for BB genotype, P<0.001), while CSN3 alleles were associated with significantly different effects on milk yield and coagulation properties; the BB genotype showed higher values for milk yield (P<0.01) and curd firmness (a30 P<0.01 and a2r P<0.01) and lower values for coagulation and curd firming time (P<0.01) than the AA genotype. Cheese made with LGB BB milk showed higher percentage protein recovery (P<0.01); cheese made with CSN3 BB milk showed higher percentage fat recovery (P<0.05)

Influence of fresh forage-based diets and αS1-casein (CSN1S1) genotype on nutrient intake and productive, metabolic, and hormonal responses in milking goats

Polymorphism at the αS1-casein locus (CSN1S1) in goats influences several milk production traits. Milk from goats carrying strong alleles, which are associated with high αS1-casein (αS1-CN) synthesis, has higher fat and casein contents, longer coagulation time and higher curd firmness than milk from goats with weak alleles linked to low αS1-CN content. Nutrition also affects these milk properties; therefore, it is important to better understand the interaction between dietary characteristics and the CSN1S1 genotype in goats. This study aimed to investigate the effect of fresh forage based diet or energy supplement on feeding behavior, milk production, and metabolic and hormonal parameters of Girgentana goats with different genotypes at CSN1S1 loci. From a group of goats genotyped by PCR at the DNA level, 12 were selected because they had the same genotype for αS2-CN, β-CN, and κ-CN but a different genotype for αS1-CN: 6 were homozygous for strong alleles at the CSN1S1 loci (AA) and 6 were heterozygous for a weak allele (AF). Goats of each genotype were allocated to 3 subgroups and fed 3 diets ad libitum in a 3×3 Latin square design. The diets were sulla (Hedysarum coronarium L.) fresh forage, sulla fresh forage plus 800g/d of barley meal (SFB), and mixed hay plus 800g/d of barley meal (MHB). Diet had a stronger effect than CSN1S1 genotype. The SFB diet led to the highest energy intake, dry matter (DM) digestibility, and milk yield. The fresh forage diets (SFF and SFB) increased DM and crude protein (CP) intake, CP digestibility, and milk CN compared with the MHB diet. The diets supplemented with energy (SFB, MHB) reduced milk fat and urea, improved CP utilization for casein synthesis, and limited body fat mobilization, in accordance with a lower level of nonesterified fatty acids and higher levels of glucose and IGF-1. With regard to CSN1S1 genotype, AA goats showed higher CP digestibility and lower free thyroxine hormone and cholesterol levels than AF goats. Significant diet × genotype interactions indicated how AA goats, compared with AF goats, showed higher DM digestibility and milk yield when fed the SFB diet, which had more energy. A reduction in free triiodothyronine hormone occurred in AF goats fed the MHB diet, whereas no differences were observed in AA goats. These results demonstrate how goats with a higher capacity for αS1-CN synthesis exhibit more efficient energy and protein utilization, evident at the digestive level, and better productive responses to high-nutrition diets

Effect of CSN1S1 G allele on some characteristics of milk produced by Cinisara cows

In bovine milk about 37-40% of total casein is represented by s1-casein. The CSN1S1 G allele is characterised by a less efficient protein synthesis (about 40%) compared with the other known alleles because of the insertion, in the 19th exon, of a Long Interspersed Nuclear Element relict. This allele is responsible for a reduced mRNA stability. The lower proportion of s1-casein in milk is associated with faster formation, higher firmness and greater resistance to compression of the curd. The purpose of this study was to analyse the G allele distribution at the CSN1S1 locus in Cinisara breed and its influence on some milk traits. The Cinisara is a small Sicilian breed for which, recently, recovery and safeguard actions were accomplished. Milk obtained from this breed is mainly used to produce the Caciocavallo Palermitano, a typical stretched-curd (pasta filata) cheese. This product represents an example of link between breed, breeding environment, and traditional dairy technology, factors highly appreciated by consumers today. We typed 192 lactating cows, by means of PCR-RFLP analyses, in order to identify CSN1S1 G carriers. Individual milk samples of 14 cows (low as1-casein) and 23 control cows (normal s1-casein) were analysed for total nitrogen (TN) and non-casein nitrogen (NCN) according to FIL-IDF standard procedures, and for fat by an infrared method (Combi-foss 6000, Foss Electric, Hillerød, Denmark). Coagulation properties were measured using the Formagraph. Control cows were chosen in the same herds of G carrier cows and had comparable lactation stage and number. Data were analysed using ANOVA procedure in which the fixed factor of CSN1S1 was evaluated. The frequency of the CSN1S1 G allele in the 192 analyzed cows was 0.039 (1 homozygous and 13 heterozygous cows). The analyses of milk traits and clotting parameters show that also in Cinisara the CSN1S1 G allele is associated with significantly lower protein (p<.049), casein (p<.046), and casein index (p<.039). Moreover the milk of cows with CSN1S1 G allele show a tendency to a lower percentage of fat (p<.072). As a consequence, the CSN1S1 G allele is associated with significantly lower coagulation time (r) (p<.019), and tendentially lower curd firming time (k20) (p<.057), and higher curd firmness (a30) (p<.063). These results confirm that CSN1S1 G allele considerably affects milk traits. Further analyses are necessary to evaluate the influence of this allele on yield and composition of Caciocavallo Palermitano cheese

Effect of β-lactoglobulin (LGB) and k-casein (CSN3) alleles on some characteristics of milk produced by Cinisara cows

It is well known that β-lactoglobulin (LGB) and k-casein (CSN3) alleles affect milk cheese making properties. The purpose of this study was to analyse the A and B allele distribution at the LGB and CSN3 loci in Cinisara breed and their influence on milk and cheese making production traits. The Cinisara is a Sicilian breed which, according to DAD-IS database (FAO), consists of about 5,000 individuals and is reared mainly for milk production which is usually transformed into Caciocavallo Palermitano, a typical stretched-curd cheese. We typed 314 lactating cows at the LGB and CSN3 loci by means of RFLP-PCR analyses. The frequencies of LGB A and B alleles were 0.22 and 0.78, respectively; the frequencies of CSN3 A and B alleles were 0.46 and 0.54, respectively. Individual milk samples were analysed for total nitrogen (TN) and non-casein nitrogen (NCN) according to FIL-IDF standard procedure. The coagulation properties were measured using the Formagraph. Data were analysed using ANOVA procedure in which the fixed effect of CSN3 genotype or the fixed effect of LGB genotype was evaluate. The results of this study show that also in Cinisara the LGB A and B alleles are associated with significantly different effects on NCN (lower for BB genotype, P<0.001), and casein index (higher value for BB genotype, P<0.0001). As a consequence, the LGB alleles are associated also with significantly different effects on coagulation properties such as coagulation time (r) and curd firming time (k20) (lower value for BB genotype, P<0.011, P<0.050, respectively); while they are not associated with different effects on curd firmness (a30 and a2r). CSN3 A and B alleles are not associated with different effects on NCN, and casein index. On the contrary, they are associated with significantly different effects on coagulation properties: BB genotype shows lower value for coagulation time (r) (P<0.0001) and curd firming time (k20) (P<0.0001); higher value for curd firmness (a30) (P<0.0001) and curd firmness (a2r) (P<0.0001) than AA genotype. Our results confirm again that LGB and CSN3 genotypes affect cheese making properties and show that Cinisara cows, due to higher frequencies of alleles with positive effect on cheese making properties, produce a high quality milk for cheese processing

Effect of CSN1S1 G allele on some characteristics of milk produced by Cinisara cows

In bovine milk about 37-40% of total casein is represented by s1-casein. The CSN1S1 G allele is characterised by a less efficient protein synthesis (about 40%) compared with the other known alleles because of the insertion, in the 19th exon, of a Long Interspersed Nuclear Element relict. This allele is responsible for a reduced mRNA stability. The lower proportion of s1-casein in milk is associated with faster formation, higher firmness and greater resistance to compression of the curd. The purpose of this study was to analyse the G allele distribution at the CSN1S1 locus in Cinisara breed and its influence on some milk traits. The Cinisara is a small Sicilian breed for which, recently, recovery and safeguard actions were accomplished. Milk obtained from this breed is mainly used to produce the Caciocavallo Palermitano, a typical stretched-curd (pasta filata) cheese. This product represents an example of link between breed, breeding environment, and traditional dairy technology, factors highly appreciated by consumers today. We typed 192 lactating cows, by means of PCR-RFLP analyses, in order to identify CSN1S1 G carriers. Individual milk samples of 14 cows (low as1-casein) and 23 control cows (normal s1-casein) were analysed for total nitrogen (TN) and non-casein nitrogen (NCN) according to FIL-IDF standard procedures, and for fat by an infrared method (Combi-foss 6000, Foss Electric, Hillerød, Denmark). Coagulation properties were measured using the Formagraph. Control cows were chosen in the same herds of G carrier cows and had comparable lactation stage and number. Data were analysed using ANOVA procedure in which the fixed factor of CSN1S1 was evaluated. The frequency of the CSN1S1 G allele in the 192 analyzed cows was 0.039 (1 homozygous and 13 heterozygous cows). The analyses of milk traits and clotting parameters show that also in Cinisara the CSN1S1 G allele is associated with significantly lower protein (p<.049), casein (p<.046), and casein index (p<.039). Moreover the milk of cows with CSN1S1 G allele show a tendency to a lower percentage of fat (p<.072). As a consequence, the CSN1S1 G allele is associated with significantly lower coagulation time (r) (p<.019), and tendentially lower curd firming time (k20) (p<.057), and higher curd firmness (a30) (p<.063). These results confirm that CSN1S1 G allele considerably affects milk traits. Further analyses are necessary to evaluate the influence of this allele on yield and composition of Caciocavallo Palermitano cheese

Early upregulation of 18-kDa translocator protein in response to acute neurodegenerative damage in TREM2-deficient mice

Mutations in the TREM2 gene confer risk for Alzheimer's disease and susceptibility for Parkinson's disease (PD). We evaluated the effect of TREM2 deletion in a 1-methyl 4-phenyl 1,2,3,6-tetrahydropyridine (MPTP)–induced PD mouse model, measuring neurodegeneration and microglia activation using a combined in vivo imaging and postmortem molecular approach. In wild-type mice, MPTP administration induced a progressive decrease of [11C]FECIT uptake, culminating at day 7. Neuronal loss was accompanied by an increase of TREM2, IL-1β, and translocator protein (TSPO) transcript levels, [11C]PK11195 binding and GFAP staining (from day 2), and an early and transient increase of TNF-α, Galectin-3, and Iba-1 (from day 1). In TREM2 null (TREM2−/−) mice, MPTP similarly affected neuron viability and microglial cells, as shown by the lower level of Iba-1 staining in basal condition, and reduced increment of Iba-1, TNF-α, and IL-1β in response to MPTP. Likely to compensate for TREM2 absence, TREM2−/− mice showed an earlier increment of [11C]PK11195 binding and a significant increase of IL-4. Taken together, our data demonstrate a central role of TREM2 in the regulation of microglia response to acute neurotoxic insults and suggest a potential modulatory role of TSPO in response to immune system deficit