Derivation of multivariate indices of milk composition, coagulation properties, and curd yield in Manchega dairy sheep.

This study approaches the interrelation patterns between composition of milk and whey, curd yield, chromaticity, syneresis, and technological quality of Manchega sheep milk using multivariate factor analysis. In addition, the effect of the main husbandry components (flock, prolificacy, season of the year, stage of lactation, and parity) on the common latent factors that define the pattern of variation of Manchega milk was assessed. For this purpose, 1,200 individual Manchega ewe milk samples from 4 different flocks registered under the Protected Designation of Origin Queso Manchego were analyzed (50 ewes/flock). Samples were collected in 2 different seasons of the year (spring and autumn) and at 3 time points per season: early, mid-, and late lactation. The obtained results suggested that curd yield mainly depends on milk composition, and the retention of water in the curd is related to coagulation traits. Thus, composition and moisture content could be useful indicators to assess the efficiency and quality of milk intended for cheesemaking, regardless of the analysis of coagulation properties. Finally, in terms of husbandry, a direct effect of flock and stage of lactation was observed on all analyzed factors, with a lower influence of season and parity.

Using multivariate analysis to explore the relationships between color, composition, hygienic quality, and coagulation of milk from Manchega sheep.

This study explores the relationships between composition, hygienic quality, and color values of milk to evaluate whether colorimetry could be used as a valid predictor of the quality of raw milk and its coagulation. For this purpose, we performed analyses in 1,200 individual samples of Manchega sheep milk from 4 flocks from the region of Castilla-La Mancha, Spain. Based on the measured variables, we determined the differences and similarities between coagulating and noncoagulating samples using discriminant analysis techniques. The variables with a higher discriminant ability were lactose content, somatic cell score, pH, and the color values lightness (L*) and red/green value (a*). The model based on color values showed a predictive ability similar to that found in the model based on milk composition and hygienic quality. Canonical correlation analysis allowed us to explore the relationships between both sets of variables. Canonical correlations for the first and second pair of canonical values were 0.794 and 0.438, respectively. Both values were significant and represented 92.82% of the observed variability. The correlation structure showed that color values had a strong correlation with fat and protein content and with total solids, and they had a weak correlation with lactose content and somatic cell score. The 2 first combinations of standardized canonical variability could be considered a predictable measure of the composition and, to a lesser extent, the hygiene of milk. Measurement of color values could be a rapid and effective means of supplementing standard analyses when determining the coagulation ability of Manchega sheep milk.

Absence of microglial CX3CR1 impairs the synaptic integration of adult-born hippocampal granule neurons.

Microglia are immune cells that play a crucial role in maintaining brain homeostasis. Among the mechanisms of communication between microglia and neurons, the CX3CL1/CX3CR1 axis exerts a central modulatory role. Animals lacking CX3CR1 microglial receptor (CX3CR1-/- mice) exhibit marked alterations not only in microglia but also in neurons located in various regions of the brain. Here we show that microglial depletion of CX3CR1 leads to the deficient synaptic integration of adult-born granule neurons in the dentate gyrus (DG), both at the afferent and efferent level. Regarding the alterations in the former level, these cells show a reduced number of dendritic spines, which also exhibit morphological changes, namely enlargement and shortening. With respect to changes at the efferent level, these cells show a reduced area of axonal terminals. Both at the afferent and efferent level, synapses show ultrastructural enlargement, but they are depleted of synaptic vesicles, which suggests impaired functionality. We also show that selective increased microglial activation and extracellular matrix deposition in the zones in which the afferent synaptic contacts of these cells occur, namely in the molecular and the granule layer of the DG. In order to evaluate the impact of these structural alterations from a functional point of view, we performed a battery of behavioral tests related to hippocampal-dependent emotional behavior. We observed that female CX3CR1-/- mice exhibit a hyperactive, anxiolytic-like and depressive-like phenotype. These data shed light on novel aspects of the regulation of adult hippocampal neurogenesis by microglia that could be highly relevant for research into mood disorders.

Multivariate analysis of the milk coagulation process in ovine breeds from Spain.

In Spain, ewe milk is mainly used for cheesemaking, and farming systems have traditionally been based on the use of autochthonous breeds. However, in recent years, the progressive introduction of highly productive foreign breeds in Spanish farms has led to an increasing interest in the characterization of dairy sheep breeds to evaluate whether genetic selection schemes should focus on productivity or milk technological aptitude. The purpose of this work was to explore milk composition and coagulation to classify 4 of the main dairy sheep breeds used in Spain. This study included 832 individual ewe milk samples from the breeds Manchega, Assaf, Merino de Grazalema, and Merino de Los Pedroches. Samples were analyzed for native pH, composition (fat, protein, lactose, and total solids), coagulation properties, and individual laboratory curd yield. An indicator of coagulation efficiency was also determined. Canonical discriminant analysis was performed to establish differences and similarities among breeds based on the measured variables. In addition, cluster analysis was performed to study and quantify the concrete relationships among the discriminated groups. Discriminant analysis proved to be a powerful tool to accurately draw distinctions between breeds. In all cases, discrimination among breeds was evident and the 4 breeds could be easily differentiated. Cluster analysis showed greater similarity between Merino de Grazalema and Assaf compared with the other breeds, and F-statistics indicated a higher discriminating ability for the variables related to milk composition. However, Merino de Grazalema and Manchega were difficult to separate according to milk composition, but the coagulation process differenced them clearly. Coagulation also evidenced similarities between Manchega and Merino de Los Pedroches, although the latter was revealed to be the most different breed of all 4, which could lay the ground for its differentiation as an independent breed in the Official Catalogue of Spanish Livestock Breeds.

Coagulation efficiency and its determinant factors: A case study for Manchega ewe milk in the region of Castilla-La Mancha, Spain.

Milk coagulation, especially in small ruminant species, is often hard to evaluate, as coagulation traits are normally considered individually and several factors related to udder health might distort yield calculation. Due to the lack of studies about these factors, our objective was to determine milk coagulation efficiency (CE) and its determinants using a deterministic technical efficiency approach, an ordinary least square regression model, and ANOVA. Milk from 300 Manchega ewes was collected and analyzed for composition, milk coagulation properties, and hygienic quality. The study results indicate that the estimated CE in Manchega ewes was 0.69, implying an important proportion of the animals produce poorly coagulating milk. The results of the ordinary least square regression model and ANOVA revealed that the main factor causing inefficiency was the initial pH of milk. Crude protein, casein and plasmin activity had moderate effects on CE, and, finally, other factors such as freezing point depression, somatic cell count, colony-forming units, and fat concentration had minor effects.

A systems approach identifies time-dependent associations of multimorbidities with pancreatic cancer risk.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is usually diagnosed in late adulthood; therefore, many patients suffer or have suffered from other diseases. Identifying disease patterns associated with PDAC risk may enable a better characterization of high-risk patients. METHODS: Multimorbidity patterns (MPs) were assessed from 17 self-reported conditions using hierarchical clustering, principal component, and factor analyses in 1705 PDAC cases and 1084 controls from a European population. Their association with PDAC was evaluated using adjusted logistic regression models. Time since diagnosis of morbidities to PDAC diagnosis/recruitment was stratified into recent (<3 years) and long term (≥3 years). The MPs and PDAC genetic networks were explored with DisGeNET bioinformatics-tool which focuses on gene-diseases associations available in curated databases. RESULTS: Three MPs were observed: gastric (heartburn, acid regurgitation, Helicobacter pylori infection, and ulcer), metabolic syndrome (obesity, type-2 diabetes, hypercholesterolemia, and hypertension), and atopic (nasal allergies, skin allergies, and asthma). Strong associations with PDAC were observed for ≥2 recently diagnosed gastric conditions [odds ratio (OR), 6.13; 95% confidence interval CI 3.01-12.5)] and for ≥3 recently diagnosed metabolic syndrome conditions (OR, 1.61; 95% CI 1.11-2.35). Atopic conditions were negatively associated with PDAC (high adherence score OR for tertile III, 0.45; 95% CI, 0.36-0.55). Combining type-2 diabetes with gastric MP resulted in higher PDAC risk for recent (OR, 7.89; 95% CI 3.9-16.1) and long-term diagnosed conditions (OR, 1.86; 95% CI 1.29-2.67). A common genetic basis between MPs and PDAC was observed in the bioinformatics analysis. CONCLUSIONS: Specific multimorbidities aggregate and associate with PDAC in a time-dependent manner. A better characterization of a high-risk population for PDAC may help in the early diagnosis of this cancer. The common genetic basis between MP and PDAC points to a mechanistic link between these conditions.

Detailed characterization of MLH1 p.D41H and p.N710D variants coexisting in a Lynch syndrome family with conserved MLH1 expression tumors.

Lynch syndrome (LS) is an autosomal dominant cancer-susceptibility disease caused by inactivating germline mutations in mismatch repair (MMR) genes. Variants of unknown significance (VUS) are often detected in mutational analysis of MMR genes. Here we describe a large family fulfilling Amsterdam I criteria carrying two rare VUS in the MLH1 gene: c.121G > C (p.D41H) and c.2128A > G (p.N710D). Collection of clinico-pathological data, multifactorial analysis, in silico predictions, and functional analyses were used to elucidate the clinical significance of the identified MLH1 VUS. Only the c.121G > C variant cosegregated with LS-associated tumors in the family. Diagnosed colorectal tumors were microsatellite unstable although immunohistochemical staining revealed no loss of MMR proteins expression. Multifactorial likelihood analysis classified c.2128A > G as a non-pathogenic variant and c.121G > C as pathogenic. In vitro functional tests revealed impaired MMR activity and diminished expression of c.121G > C. Accordingly, the N710 residue is located in the unconserved MLH1 C-terminal domain, whereas D41 is highly conserved and located in the ATPase domain. The obtained results will enable adequate genetic counseling of c.121G > C and c.2128A > G variant carriers and their families. Furthermore, they exemplify how cumulative data and comprehensive analyses are mandatory to refine the classification of MMR variants.

Selenium status in a group of schoolchildren from the region of Madrid, Spain.

BACKGROUND: To assess the selenium status of a group of schoolchildren from the Region of Madrid, Spain. METHODS: Study subjects comprised 483 children (216 boys and 267 girls) aged between 8 and 13 years. Selenium intake was determined using a 3-day food record. The foods consumed were transformed into energy and nutrients, and the selenium intake was compared with that recommended. Serum selenium levels were also recorded. RESULTS: Mean (SD) selenium intake [91.0 (25.2) µg day(-1) ] was above the recommended level in 99.4% of subjects; the main dietary sources were cereals, meats, fish and milk products. The serum selenium concentration [mean (SD) 71.1 (14.4) µg L(-1) ], however, was <60 µg L(-1) in 13.9% of subjects, and <45 µg L(-1) in 5.6%. The serum selenium concentration correlated with the selenium intake (r = 0.169; P < 0.05). Children with a serum selenium concentration of <75 µg L(-1) had significantly smaller selenium intakes than those with a serum concentration of ≥75 µg L(-1) . CONCLUSIONS: Although selenium intake was generally above that recommended, the serum selenium concentration of the children could be improved. This could be achieved by increasing the relative consumption of cereals and other selenium rich foods such as fish.

Relationship between Paratuberculosis and the microelements Copper, Zinc, Iron, Selenium and Molybdenum in Beef Cattle.

To study the deficiency of minerals and its relationship with Paratuberculosis, blood, serum, and fecal samples were obtained from 75 adult bovines without clinical symptoms of the disease and from two bovines with clinical symptoms of the disease, from two beef herds with a previous history of Paratuberculosis in the Province of Buenos Aires, Argentina. Serum samples were processed by ELISA and feces were cultured in Herrolds medium. Copper, zinc and iron in serum were quantified by spectrophotometry and selenium was measured by the activity of glutathione peroxidase. We also determined copper, zinc, iron and molybdenum concentrations in pastures and the concentration of sulfate in water. Mycobacterium avium subsp paratuberculosis (Map) was isolated from 17.3% of fecal samples of asymptomatic animals and from the fecal samples from the two animals with clinical symptoms. All the Map-positive animals were also ELISA-positive or suspect, and among them, 84.6% presented low or marginal values of selenium and 69.2% presented low or marginal values of copper. The two animals with clinical symptoms, and isolation of Map from feces and organs were selenium-deficient and had the lowest activity of glutathione peroxidase of all the animals from both herds. All the animals negative to Map in feces and negative to ELISA had normal values of Se, while 13.8% of animals with positive ELISA or suspect and culture negative presented low levels of Se. Half of the animals that were negative both for ELISA and culture in feces were deficient in copper but none of them presented low values of selenium. The content of molybdenum and iron in pasture was high, 2.5 ppm and 1.13 ppm in one herd and 2.5 ppm and 2.02 ppm in the other, respectively, whereas the copper:molybdenum ratio was 1.5 and 5.2, respectively. These results do not confirm an interaction between imbalances of the micronutrients and clinical Paratuberculosis, but show evidence of the relationship between selenium deficiencies in animals with Map infection and ELISA positive results.

Optimization of antimicrobial treatment in sepsis.

Sepsis represents a serious risk to the life of any patient, which is why it is crucial to start an effective treatment in all its extremes as soon as possible, that is, the chosen antibiotics must have activity against the pathogen that produces the condition and, in addition, they must be dosed considering the patient's situation in all its extremes. It should be considered that it will be necessary to adjust the dose when there is edema (drugs with reduced volume of distribution), hypoproteinemia (drugs bound to proteins in a high proportion), obesity, and also when they require the use of external techniques such as ECMO or any of the different types of hemodialysis and hemofiltration.

Pharmacokinetics/Pharmacodynamics and tolerability of cefiderocol in the clinical setting.

Cefiderocol is a new cephalosporin with a catechol in its chemical structure faciliting its access to the interior of bacteria through iron channels. In addition, it is broadly stable to beta-lactamases. The pharmacokinetic profile is a beta-lactam one: no oral absorption, and with a wide distribution within the vascular space and the interstitial fluid of well vascularized tissues, reaching therapeutic concentrations in the alveolar lavage fluid and within the macrophage. The binding of cefiderocol to human plasma proteins, primarily albumin, is moderate (range 40-60%). The terminal elimination half-life in healthy adult subjects was 2 to 3 hours. Cefiderocol is mainly renally eliminated, so dose adjustments are recommended in subjects with moderate / severe renal impairment, in case of dialysis, and probably in patients with external clearance. Like other beta-lactams, the PK / PD parameter that has been shown to best correlate with efficacy is the efficacy time of unbound plasma concentrations (%fT>MIC), which must be close to 100% to achieve a bactericidal effect. This is possible with 2 g in a 3-hour infusion every 8 hours. In controlled trials appears to be well tolerated, similar to comparators: meropenem or imipenem-cilastatin. Cefiderocol has no apparent clinically significant effect on ECG parameters nor on plasma iron values.

Identification of genes associated with environmental persistence in Campylobacter jejuni and Campylobacter coli isolates from processing in a broiler abattoir.

The aim of this study was to determine the prevalence of the htrA, htrB and ppk1 genes -all of which are related to environmental persistence- in C. jejuni and C. coli isolates obtained from abattoir samples at the arrival of broilers (initial stage) and in meat products after processing (final stage). A total of 119 DNA extracts (55 C. jejuni and 64 C. coli) were included in the study. Identification of genes was performed by conventional PCR (one for each gene). The overall prevalence was 40.3%, 93.3% and 68.9% for the htrA, htrB and ppk1 genes, respectively. Statistically significant differences were found (p < 0.05) between prevalence of C. jejuni and C. coli for all three genes. In C. coli the prevalence was significantly higher for the htrA (p = 0.007) and htrB (p = 0.015) genes, while ppk1 gene prevalence was significantly higher in C. jejuni (p < 0.001). In addition, statistically significant increase in the frequency of htrA (p = 0.007) and htrB (p = 0.013) genes in the final product compared to broilers on arrival at the abattoir was observed in C. jejuni, but not in C. coli. These results suggest that htrA and htrB genes are involved in environmental persistence of Campylobacter jejuni.

Quantification of lentiviral vector copy numbers in individual hematopoietic colony-forming cells shows vector dose-dependent effects on the frequency and level of transduction.

Lentiviral vectors are effective tools for gene transfer and integrate variable numbers of proviral DNA copies in variable proportions of cells. The levels of transduction of a cellular population may therefore depend upon experimental parameters affecting the frequency and/or the distribution of vector integration events in this population. Such analysis would require measuring vector copy numbers (VCN) in individual cells. To evaluate the transduction of hematopoietic progenitor cells at the single-cell level, we measured VCN in individual colony-forming cell (CFC) units, using an adapted quantitative PCR (Q-PCR) method. The feasibility, reproducibility and sensitivity of this approach were tested with characterized cell lines carrying known numbers of vector integration. The method was validated by correlating data in CFC with gene expression or with calculated values, and was found to slightly underestimate VCN. In spite of this, such Q-PCR on CFC was useful to compare transduction levels with different infection protocols and different vectors. Increasing the vector concentration and re-iterating the infection were two different strategies that improved transduction by increasing the frequency of transduced progenitor cells. Repeated infection also augmented the number of integrated copies and the magnitude of this effect seemed to depend on the vector preparation. Thus, the distribution of VCN in hematopoietic colonies may depend upon experimental conditions including features of vectors. This should be carefully evaluated in the context of ex vivo hematopoietic gene therapy studies.

Coupling-barrier and non-parabolicity effects on the conduction electron cyclotron effective mass and Landé [Formula: see text] factor in GaAs double quantum wells.

We have performed a theoretical study of the effects of the non-parabolicity and coupling barrier in between GaAs quantum wells on the conduction electron cyclotron effective mass and Landé [Formula: see text] factor under the action of a growth-direction applied magnetic field. Numerical calculations are performed within the effective mass approximation and taking into account the non-parabolicity effects for the conduction-band electrons, by means of the Ogg-McCombe effective Hamiltonian. The system consists of two GaAs quantum wells connected by a Ga(1 - x)Al(x)As barrier and surrounded by Ga(1 - y)Al(y)As material. We have found that both the [Formula: see text] factor and the cyclotron effective mass are sensitive to the coupling strength, that is the height and width of the barrier in between the GaAs quantum wells. This behavior is similar for every Landé [Formula: see text] factor and the cyclotron effective mass calculated for different Landau levels. It is noticeable that the splitting between the [Formula: see text] and [Formula: see text] cyclotron effective mass increases with the central barrier width and the growth-direction applied magnetic field. As in a single quantum well, we found that the electron Landé [Formula: see text] factor increases with the growth-direction applied magnetic field, comparing quite well with the experimental reports, and that the magnetic field plays an important role in decoupling the quantum wells of the system. Additionally, we have studied the electron cyclotron effective mass and Landé g factor as functions of the Landau levels, depending on the non-parabolicity. From this result one can infer that their population must be taken into account for a complete study of the band parameters as has been proposed in previous works. The present theoretical results are in very good agreement with previous experimental reports in the limiting geometry of a single quantum well.

Molecular basis of colorrectal cancer: towards an individualized management?

Colorectal cancer (CRC) has become a highly relevant condition nowadays. In this respect, advances in the understanding of its molecular basis are key for an adequate management. From the time when the adenoma-carcinoma sequence was formulated as a carcinogenesis model to this day, when -among other things- three major carcinogenic pathways have been identified, the CRC concept has evolved from that of a single disease to the notion that each CRC is a differentiated condition in itself. The suppressor or chromosome instability pathway, the mutator or microsatellite instability pathway, and the methylator or CpG island methylation pathway allow various phenotypes to be identified within CRC. Similarly, the presence of different changes in certain genes confers several behaviors on CRC from both the prognostic and responsive standpoints to specific therapies. However, this apparent complexity does help develop the clinical management of this disease through the identification of novel, more specific therapy targets, and also markers for various behaviors within the condition, which will most likely lead us to an individualized management for these patients.

Gallbladder disease and pancreatic cancer risk: a multicentric case-control European study.

BACKGROUND AND AIMS: The overall evidence on the association between gallbladder conditions (GBC: gallstones and cholecystectomy) and pancreatic cancer (PC) is inconsistent. To our knowledge, no previous investigations considered the role of tumour characteristics on this association. Thus, we aimed to assess the association between self-reported GBC and PC risk, by focussing on timing to PC diagnosis and tumour features (stage, location, and resection). METHODS: Data derived from a European case-control study conducted between 2009 and 2014 including 1431 PC cases and 1090 controls. We used unconditional logistic regression models to estimate odds ratios (ORs) and corresponding 95% confidence intervals (CIs) adjusted for recognized confounders. RESULTS: Overall, 298 (20.8%) cases and 127 (11.6%) controls reported to have had GBC, corresponding to an OR of 1.70 (95% CI 1.33-2.16). The ORs were 4.84 (95% CI 2.96-7.89) for GBC diagnosed <3 years before PC and 1.06 (95% CI 0.79-1.41) for ≥3 years. The risk was slightly higher for stage I/II (OR = 1.71, 95% CI 1.15-2.55) vs. stage III/IV tumours (OR = 1.23, 95% CI 0.87-1.76); for tumours sited in the head of the pancreas (OR = 1.59, 95% CI 1.13-2.24) vs. tumours located at the body/tail (OR = 1.02, 95% CI 0.62-1.68); and for tumours surgically resected (OR = 1.69, 95% CI 1.14-2.51) vs. non-resected tumours (OR = 1.25, 95% CI 0.88-1.78). The corresponding ORs for GBC diagnosed ≥3 years prior PC were close to unity. CONCLUSION: Our study supports the association between GBC and PC. Given the time-risk pattern observed, however, this relationship may be non-causal and, partly or largely, due to diagnostic attention and/or reverse causation.

Different responses to doxorubicin-induced chromosome aberrations in Brazilian deer species.

The tendency toward chromosome fragility is one of the theories that may explain chromosome variation in brocket deer species (genus Mazama). We tested doxorubicin as an inducer of chromosome aberrations in lymphocytes of three brocket deer species, Mazama gouazoubira, M. americana and M. nana, compared to the marsh deer, Blastocerus dichotomus. Doxorubicin, at a concentration of 0.25 microg/mL, induced chromosome aberrations and fragile sites in all four species; the highest frequencies were seen in M. gouazoubira; they were lowest in B. dichotomus and intermediate in M. americana and M. nana. These results were expected based on previous karyotypic studies, but they failed to explain the higher sensitivity seen in M. gouazoubira. This may be because not all the aberrations and fragile sites are related to chromosome evolution in brocket deer; other factors, such as environmental influences, may be involved in chromosome fragility.