Traditional sexing methods and external egg characteristics combination allow highly accurate early sex determination in an endangered native turkey breed.

Early sex determination methods are not only crucial in the worldwide massive poultry industry, but also for small-holder producers. The profitability of sexing techniques must be accounted for when aiming to boost management, nutrition, and conservation practices in endangered poultry breeds. This becomes pivotal when the local breed dealt with belongs to an understudied species, such as the turkey. So, the main objective of this study is to identify which method combination may report a higher likelihood of successful sex determination in poults across the three-pattern varieties of the Andalusian turkey breed. A total of 84 one to two days old Andalusian turkey poults (42 black, 28 black-roan, and 14 bronze-roan) were evaluated in this study. Sex determination was performed using 15 methods, which included testing external egg metrics and eggshell color, poult morphological appraisal and phaneroptics, and behavioral traits. Possible differences across plumage varieties and the interaction between sex and plumage were observed when external egg quality was measured. Sex determination through behavioral methods in black base feathered (black and black-roan) male sex individuals showed seven times higher sensitivity when compared to the rest of the studied individuals (χ2 = 7.14, df = 1, P < 0.01). In contrast, for the black-roan plumage females, the method based on the color of down feathers was approximately four times more sensitive (χ2 = 3.95, df = 1, P ≤ 0.05). For the bronze-roan pattern, none of the sexing techniques was reported to efficiently predict sex itself. However, the most proper method combination to determine sex, independent of plumage color, was physical external egg characteristics, the color of down feathers, and behavioral approaches ("English method" and "slap technique"). The specificity values were found to be 49.12, 93.33, and 100%, while the sensitivity values were observed to be 74.64, 91.03, and 100%, which translated into accuracy of 63.10, 92.26, and 100% in black, black-roan, and bronze-roan poults, respectively. Our results suggest that the method combination tested in this study could be considered a highly accurate, simple, and affordable alternative for sex determination in turkeys. This could mean a pivotal advance for small producers of turkeys, as early sex detection can help to plan timely conservational management strategies, which is of prominent importance in the context of endangered poultry breeds.

Authentication of fattening diet of Iberian pigs according to their volatile compounds profile from raw subcutaneous fat.

The composition of volatile components of subcutaneous fat from Iberian pig has been studied. Purge and trap gas chromatography-mass spectrometry has been used. The composition of the volatile fraction of subcutaneous fat has been used for authentication purposes of different types of Iberian pig fat. Three types of this product have been considered, montanera, extensive cebo and intensive cebo. With classification purposes, several pattern recognition techniques have been applied. In order to find out possible tendencies in the sample distribution as well as the discriminant power of the variables, principal component analysis was applied as visualisation technique. Linear discriminant analysis (LDA) and soft independent modelling by class analogy (SIMCA) were used to obtain suitable classification models. LDA and SIMCA allowed the differentiation of three fattening diets by using the contents in 2,2,4,6,6-pentamethyl-heptane, m-xylene, 2,4-dimethyl-heptane, 6-methyl-tridecane, 1-methoxy-2-propanol, isopropyl alcohol, o-xylene, 3-ethyl-2,2-dimethyl-oxirane, 2,6-dimethyl-undecane, 3-methyl-3-pentanol and limonene.

New models for donor-recipient matching in lung transplantations.

OBJECTIVE: One of the main problems of lung transplantation is the shortage of organs as well as reduced survival rates. In the absence of an international standardized model for lung donor-recipient allocation, we set out to develop such a model based on the characteristics of past experiences with lung donors and recipients with the aim of improving the outcomes of the entire transplantation process. METHODS: This was a retrospective analysis of 404 lung transplants carried out at the Reina Sofía University Hospital (Córdoba, Spain) over 23 years. We analyzed various clinical variables obtained via our experience of clinical practice in the donation and transplantation process. These were used to create various classification models, including classical statistical methods and also incorporating newer machine-learning approaches. RESULTS: The proposed model represents a powerful tool for donor-recipient matching, which in this current work, exceeded the capacity of classical statistical methods. The variables that predicted an increase in the probability of survival were: higher pre-transplant and post-transplant functional vital capacity (FVC), lower pre-transplant carbon dioxide (PCO2) pressure, lower donor mechanical ventilation, and shorter ischemia time. The variables that negatively influenced transplant survival were low forced expiratory volume in the first second (FEV1) pre-transplant, lower arterial oxygen pressure (PaO2)/fraction of inspired oxygen (FiO2) ratio, bilobar transplant, elderly recipient and donor, donor-recipient graft disproportion requiring a surgical reduction (Tailor), type of combined transplant, need for cardiopulmonary bypass during the surgery, death of the donor due to head trauma, hospitalization status before surgery, and female and male recipient donor sex. CONCLUSIONS: These results show the difficulty of the problem which required the introduction of other variables into the analysis. The combination of classical statistical methods and machine learning can support decision-making about the compatibility between donors and recipients. This helps to facilitate reliable prediction and to optimize the grafts for transplantation, thereby improving the transplanted patient survival rate.

Second complete remission induced by cyclophosphamide plus bevacizumab in two patients with heavily pre-treated ovarian cancer.

During the last decade we have been developing new therapeutic strategies for the treatment of ovarian cancer, based on the best knowledge of the molecular biology. New drugs like bevacizumab are showing antiangiogenic antitumour activity in ovarian cancer in preclinical models and in clinical trials. Bevacizumab is a monoclonal human antibody that has showed activity in both monotherapy and in combination with other therapies. We present the outcome of two patient cases with recurring heavily pre-treated ovarian cancer that were treated with cyclophosphamide and bevacizumab in combination and showed a complete remission of disease. After interruption of the treatment with bevacizumab, both patients had a disease relapse. Despite poor prognosis of the disease, in both cases a second complete and maintained remission was achieved, again with the same regimen.

Rescue of Discarded Grafts for Liver Transplantation by Ex Vivo Subnormothermic and Normothermic Oxygenated Machine Perfusion: First Experience in Spain.

BACKGROUND: Ex vivo machine perfusion (MP) has been reported as a possibly method to rescue discarded organs. The main aim of this study was to report an initial experience in Spain using MP for the rescue of severely marginal discarded liver grafts, and to, secondarily, define markers of viability to test the potential applicability of these devices for the real increase in the organ donor pool. METHODS: The study began in January 2016. Discarded grafts were included in a research protocol that consisted of standard retrieval followed by 10 hours of cold ischemia. Next, either normothermic (NMP) or controlled subnormothermic (subNMP) rewarming was chosen randomly. Continuous measurements of portal-arterial pressure and resistance were screened. Lactate, pH, and bicarbonate were measured every 30 minutes. The perfusion period was 6 hours, after which the graft was discarded and evaluated as potentially usable, but never implanted. Biopsies of the donor and at 2, 4, and 6 hours after ex vivo MP were obtained. RESULTS: A total of 4 grafts were included in the protocol. The first 2 grafts were perfused by NMP and grafts 3 and 4 by subNMP. The second and third grafts showed a clear trend toward optimal recovery and may have been used. Lactate dropped to levels below 2.5 mmol/L with stable arterial and portal pressure and resistance. Clear biliary output started during MP. Biopsies showed an improvement of liver architecture with reduced inflammation at the end of the perfusion. CONCLUSION: This preliminary experience has demonstrated the potential of MP devices for the rescue of severely marginal liver grafts. Lactate and biliary output were useful for viability testing of the grafts. The utility of NMP or subNMP protocols requires further research.

Differentiation of certified brands of origins of Spanish white wines by HS-SPME-GC and chemometrics.

A headspace solid-phase microextraction gas-chromatographic (HS-SPME-GC) procedure was used to determine the composition of the volatile fraction of white wine samples from several Spanish certified brands of origin (CBO). The compounds present were previously identified by gas chromatography-mass spectrometry (GC-MS) and quantitative determinations were carried out by GC-FID. Four CBO, Rueda, Ribeiro, Penedés, and Condado de Huelva, were studied. Rueda wines present the highest concentrations of ethyl acetate (55.86-125.27 microg mL(-1)), isoamyl acetate (0.91-6.72 microg mL(-1)), hexyl acetate (0.09-0.81 microg g mL(-1)), and 2-phenethyl acetate (0.14-0.66 microg mL(-1)). Compounds such as ethyl hexanoate (0.88-2.15 microg mL(-1)) and ethyl decanoate (0.29-0.96 microg mL(-1)) appeared in higher concentration in Ribeiro, Rueda, and Penedés samples. According to the results obtained and by applying pattern-recognition procedures differentiation of the considered CBO was attained. Principal-component analysis (PCA), linear discriminant analysis (LDA), and multilayer perceptrons neural networks (MLP-NN) were used as chemometric tools for pattern-recognition studies.

Differentiation of green, white, black, Oolong, and Pu-erh teas according to their free amino acids content.

In this paper, the differentiation of green, black, Oolong, white, and Pu-erh teas has been carried out according to their free amino acid contents. Alanine, arginine, asparagine, aspartic acid, glutamic acid, isoleucine, histidine, leucine, phenylalanine, serine, theanine, threonine, and tyrosine have been determined by liquid chromatography with derivatization with o-phthalaldehyde and fluorescence detection. The chromatographic separation was achieved with a Hypersil ODS column and gradient elution. The amino acid contents were used as chemometric descriptors for classification purposes of different tea varieties. Principal component analysis, k-nearest neighbors, linear discriminant analysis, and artificial neural networks were applied to differentiate tea varieties. Using back-propagation multilayer perceptron artificial neural networks, 100% success in the classification was obtained. The most differentiating amino acids were glutamic acid, asparagine, serine, alanine, leucine, and isoleucine.

Fluorometric determination of mixtures of quinolones by means of partial least squares and neural networks.

Multivariate calibration methods (partial least squares calibration, back propagation multilayer perceptrons networks, radial basis functions and generalized regression neural networks) were applied to the simultaneous fluorometric quantification of levofloxacin, garenoxacin and grepafloxacin, without previous separation steps. A data matrix was obtained by registering the emission spectra of mixtures of the three quinolones in urine (with concentrations ranging over 0.00-0.40 microg mL-1 for each quinolone) with a 283 nm excitation at pH 4.0. The generalized regression neural network model proved to be the most adequate model for simultaneous quantification of the three quinolones in urine samples.

Some practical considerations for linearity assessment of calibration curves as function of concentration levels according to the fitness-for-purpose approach.

Since linear calibration is mostly preferred for analytical determinations, linearity in the calibration range is an important performance characteristic of any instrumental analytical method. Linearity can be proved by applying several graphical and numerical approaches. The principal graphical criteria are visual inspection of the calibration plot, the residuals plot, and the response factors plot, also called sensitivity or linearity plot. All of them must include confidence limits in order to visualize linearity deviations. In this work, the graphical representation of percent relative errors of back-calculated concentrations against the concentration of the calibration standards is proposed as linearity criterion. This graph considers a confidence interval based on the expected recovery related to the concentration level according to AOAC approach. To illustrate it, four calibration examples covering different analytical techniques and calibration situations have been studied. The proposed %RE graph was useful in all examples, helping to highlight problems related to non-linear behavior such as points with high leverage and deviations from linearity at the extremes of the calibration range. By this way, a numerical decision limit which takes into account the concentration of calibration standards can be easily included as linearity criterion in the form of %RETh=2·C-0.11. Accordingly, this %RE parameter is accurate for the decision-making related to linearity assessment according to the fitness-for-purpose approach.

[Outcomes evaluation after the implementation of a pre-hospital thrombolysis protocol in rural areas]. / Evaluación de resultados tras la implantación de un protocolo de fibrinólisis extrahospitalaria en zonas rurales.

INTRODUCTION: The aim is to evaluate the outcomes obtained from the implementation of a pre-hospital thrombolysis protocol in 3 rural emergency care teams, as well as delays and strategies of reperfusion applied in the treatment of the ST-segment elevation myocardial infarction. MATERIAL AND METHODS: Retrospective cohort study (n=52) with historical control (n=20) of the patients assisted for ST-segment elevation myocardial infarction. Medical emergency care teams, hospital, computerized medical history and ARIAM register reports were revised, obtaining epidemiological and clinical features, off-hospital management, reperfusion, time intervals and mortality. RESULTS: The baseline features in both groups were not significantly different. There was a non-significant improvement of emergency care teams-hospital diagnostic concordance (85.3 versus 76.9%). We found a similar use of nitroglycerin, morphine and aspirin; significant increase (P<0.0001) of clopidogrel/prasugrel (55 versus 90.4%) and enoxaparin/fondaparinux (35 versus 76.9%), as well as pre-hospital thrombolysis (5 versus 30,8%, P<0.03), that was applied within the first 2h to 71.4%, with a median door-needle of 40min, whereas in-hospital thrombolysis and primary angioplasty were performed after 3h from the symptoms onset (P<0.01). Delays are associated with the patient's own lateness (P<0.02). Pharmaco-invasive strategy increases (62.5 versus 84.6%) more than primary angioplasty (15 versus 17.3%), reducing in-hospital thrombolysis (35 versus 19.2%), all of them non-significant. Complications are similar and one-year mortality is reduced (P<0.67). CONCLUSIONS: The protocol is effective, safe, and reliable. It reduces delays and improves pre-hospital attention. The pharmaco-invasive strategy is a valid option.

Short-term evolution of renal transplant with grafts from donation after cardiac death: Type III Maastricht category.

BACKGROUND: Kidney transplantation from donors after cardiac death (Type III Maastricht category) is a therapeutic option for patients with terminal renal failure. MATERIALS AND METHODS: We present a cohort of 8 patients who received a kidney transplant from donors after cardiac death (DCD). We analyzed the analytical results for the first 6 months after transplantation. RESULTS: We included 8 cases of kidney transplants with organs from DCD (Type III Maastricht category). The mean age of donors was 58.40 ± 4.39 years and 3 (60%) were male. The mean creatinine (Cr) level prior to death was 1.10 ± 0.36 mg/dL. The mean age of recipients was 59.88 ± 10.58 years and 7 (87.5%) were male. Seven patients (87.5%) were on hemodialysis, whereas only 1 (12.5%) was on peritoneal dialysis. The median time on renal replacement therapy was 18 months (range, 2-76). Mean total warm ischemia time (WIT) was 24.88 ± 6.72 minutes, whereas the mean real WIT was 20.13 ± 4.51 minutes. The mean cold ischemia time (CIT) was 6 hours and 12 minutes ± 2 hours. Preimplantation biopsy showed acute tubular necrosis (extensive 40%). Tubular atrophy was mild in 100% of cases. After transplantation, 6 patients (75%) had delayed graft function requiring dialysis sessions whereas 2 patients (25%) did not require renal replacement therapy. Mean Cr level at 1, 3, and 6 months after transplantation was 2.37, 1.75, and 1.17 mg/dL, respectively. CONCLUSION: Kidney transplantation with grafts from donors after cardiac arrest Maastricht Type III evolves favorably in the short term. According to preliminary results, controlled asystole donation could be an effective alternative to transplantation.

[Systemic capillary leak syndrome: hypoalbuminemia, hemoconcentration and shock. Presentation of a case]. / Síndrome de fuga capilar sistémica: hipoalbuminemia, hemoconcentración y shock. A propósito de un caso.

Systemic capillary leak syndrome is a rare disorder of unknown etiology and often recurrent episodes characterized by increased capillary permeability that allows a leakage of fluid and proteins from the circulatory system to the interstitial space leading to shock and massive edema. The lack of recognition of this disease may be due to its unespecific signs and symptons of presentation, its rapid clinical progression and high mortality of the acute episodes. General physicians are usually the first to evaluate patients with this kind of disorder, either in the pre-hospital situation, hospital emergency units or even (in the milder cases) in the health centers. Its poor outcome and the improvement in the prognosis, if appropriate treatment is initiated, leads us to emphasize the importance of recognizing this pathology in order to start the appropriate intensive care and emergency treatment.

Characterization and quantification of 4-methylsterols and 4,4-dimethylsterols from Iberian pig subcutaneous fat by gas chromatography-mass spectrometry and gas chromatography-flame ionization detector and their use to authenticate the fattening systems.

4-Methylsterols and 4,4-dimethylsterols of 47 samples of subcutaneous fat from Iberian pigs reared on two different fattening systems, "Extensive" and "Intensive", have been analyzed by GC-MS and GC-FID. The lipids were extracted by melting the subcutaneous fat in a microwave oven. The unsaponifiable matter was fractionated by thin layer chromatography. Then, the analysis was performed on a capillary SPB-5 column (30 m × 0.25 mm i.d., 0.15 µm film thickness), with hydrogen as a carrier gas and using a flame ionization detector. n-eicosanol was used as internal standard for quantification of individual methylsterols. These compounds have been analyzed by GC-MS for their identification. The full scan of free and trimethyl silyl ethers was used as acquisition mode. Six compounds have been identified for the first time in this type of samples: (3ß,4α,5α)-4-methyl-cholesta-7-en-3-ol, (3ß,4α,5α)-4-methyl-cholesta-8(14)-en-3-ol, (3ß,5α)-4,4-dimethyl-cholesta-8(14),24-dien-3-ol, (3ß)-lanosta-8,24-dien-3-ol, (3ß, 5α)-4,4-dimethyl-cholesta-8,14-dien-3-ol and (3ß)-lanost-9(11),24-dien-3-ol. The samples were derivatized as trimethyl silyl ethers before their analysis by GC-FID. By using these compounds as chemical descriptors, pattern recognition techniques were applied to differentiate between extensive and intensive fattening systems of Iberian pig. Several pattern recognition techniques, such as principal component analysis, linear discriminant analysis, support vector machines, artificial neural networks, soft independent modeling of class analogy and k nearest neighbors, have been used in order to find out a suitable classification model. A multilayer perceptron artificial neural network based on the contents of the above mentioned compounds allowed the differentiation of the two fattening systems with an overall classification performance of 91.7%.

Outcomes in renal transplantation with expanded-criteria donors.

BACKGROUND: Kidney transplantation is the treatment of choice for patients with end-stage renal disease. In recent years donor criteria have changed to increase the percentage of expanded-criteria donors (ECDs). The aim of this study was to analyze transplants from ECDs obtained at our institution from. 2010 to 2012. We studied the comorbidity of ECD, preimplantation histologic study, renal function, and survival of transplanted grafts. PATIENTS AND METHODS: Eighty ECDs (160 kidneys) were analyzed. Forty-nine grafts were not implanted owing to macroscopic lesions (37 kidneys) or histologic findings on preimplantation biopsy (12 kidneys). Finally, 60 grafts from ECDs were implanted in our center. We analyzed the characteristics of the grafts (kidney function, creatinine clearance) and compared the data with a control group of allografts from standard-criteria donors (n = 14). RESULTS: The median age of the ECD group was 72 years (range 65-77). No differences were found in certain characteristics between the ECDs whose kidneys were or were not implanted (hypertension, diabetes, creatinine at the time of the donation or proteinuria). However, there were differences in donor age (75 vs 67; P = .043), increased preimplantation biopsy score (6.8 ± 1.3 vs 4.8 ± 1.1; P = .041), and a higher percentage of cardiovascular disease (62.5% vs 43%; P = .038). Comparison of ECD and non-ECD grafts showed a lower creatinine clearance at 1 year (50 ± 05 mL/min vs 69 ± 96 mL/min, respectively; P < .001) and 2 years (50 ± 07 mL/min vs 67 ± 74 mL/min; P < .001) after transplantation. There were no differences in delayed graft function or graft survival between the 2 groups at 2 years after transplantation (95% vs 100%; P = .38). CONCLUSIONS: We found no differences in graft survival from ECD compared with the control group of standard-criteria donors. The evaluation of grafts from ECD may be a strategy to increase the number of kidney transplants.

[Arreflexic coma and MELAS syndrome]. / Coma arrefléxico en el síndrome de MELAS.

MELAS is a progressive neurodegenerative and fatal disease characterized by mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes. It is the result of a mitochondrial DNA mutation. Although the incidence of MELAS is currently unknown, it is suspected that approximately 1 out of every 5,000 persons world-wide have some type of defect in mitochondrial DNA. Cardinal clinical features observed in more than 90% of the patients include severe headache that may be associated with stroke-like episodes, seizures and the onset of symptoms before the age of 40 years. Diagnosis is established through genetic test or by with muscle biopsies that reveal the presence of ragged-red fibers. Prognosis is poor, with death at an early age. In this article, we present the clinical case of a 31-year old women diagnosed of MELAS syndrome who was admitted to the Intensive Care Unit of our hospital with arreflexic coma.

Characterization of aniseed-flavoured spirit drinks by headspace solid-phase microextraction gas chromatography-mass spectrometry and chemometrics.

The volatile composition of aniseed-flavoured spirit drinks was studied by headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS). The influence of the time, temperature, volume of sample and ionic strength on the extraction were considered. Several aniseed-flavoured spirit drinks, such as pastis, sambuca, anis and raki were analyzed. The major compounds found were trans-anethole (41.22-98%), cis-anethole (0.77-18.65%) and estragole (0.1-17.96%). gamma-Himachalene (0-28.07%) and alpha-himachalene (0-4.8%) were also present in anis and raki beverages. The compounds determined were used as chemical descriptors to differentiate the four types of beverages considered. Principal component analysis (PCA), linear discriminant analysis (LDA) and artificial neural networks (ANN) were used as chemometric tools for characterization purposes.

Erythema figuratum in septic babesiosis.

Babesiosis is a rare worldwide-distributed protozoal zoonosis caused by a haemoprotozoan of the genus Babesia, transmitted through bites of tick of the genus Ixodes. The first demonstrated case of human babesiosis in the world was discovered in Europe, in 1957. However, most of the cases were reported later in the north-east of the United States where Babesia microti has been the cause of over 300 cases of human babesiosis since 1969. In Europe, the most severe cases are observed in asplenic patients infected by a parasite of cattle, the Babesia divergens. Only two cases of babesiosis have been reported in Spain. We present a case of erythema figuratum associated to septic babesiosis in a non-splenectomized man, which is currently the third case of babesiosis in Spain.

Classification of aniseed drinks by means of cluster, linear discriminant analysis and soft independent modelling of class analogy based on their Zn, B, Fe, Mg, Ca, Na and Si content.

Zinc, boron, iron, magnesium, calcium, sodium and silicon were determined in aniseed drinks by inductively coupled plasma-atomic emission spectrometry (ICP-AES). These elements were considered as chemical descriptors to characterise Spanish-certified aniseed drinks brands of origin. Different pattern recognition (PR) procedures were applied for these purposes. This chemometric study have included methods like principal component analysis (PCA), non-supervised PR techniques such as cluster analysis (CA), supervised PR methods of hard modelling like linear discriminant analysis (LDA) and soft independent modelling of class analogy (SIMCA).