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OBJECTIVE: To analyse compliance with the recommendations on the insertion-maintenance of peripheral venous catheters (PVC) and the incidence of complications according to the healthcare department that inserted the PVCs. PATIENTS AND METHODS: We conducted a prospective cohort follow-up study of PVCs, from their insertion in the emergency or internal medicine (IM) department until their withdrawal. RESULTS: We monitored 590 PVCs, 274 from the emergency department and 316 from IM. In terms of compliance with the process indicators, there was a cannulation rate in the antecubital fossa of 3.5 and 1.6 per 100 catheters-day (P<.001) in the emergency and IM departments, respectively. The sterile placement rates were 1.6 and 12.4 (P<.001), and the rate for transparent dressing was 2.1 and 11.5 (P<.001) per 100 catheters-day in the emergency and IM departments, respectively. The complications rates showed no differences between the departments. The most common complication was phlebitis (95 cases, 16.1%). CONCLUSIONS: Compliance with the insertion-maintenance recommendations for PVC showed differences between the departments; however, the incidence of complications was similar.
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In this Letter, we present the design, simulation (2D and 3D), fabrication, and experimental characterization of compact and fully etched focusing gratings for a horizontal slot waveguide based on a silicon nitride layer sandwiched between amorphous silicon and a silicon-on-insulator. The measured coupling losses are about 4 dB with a 3 dB bandwidth of 38 nm. The fully etched configuration allows the fabrication in a single lithography step.
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BACKGROUND: Identification of the best embryos to transfer is a key element for success in assisted reproduction. In the last decade, several morphological criteria of oocytes and embryos were evaluated with regard to their potential for predicting embryo viability. The introduction of polarization light microscopy systems has allowed the visualization of the meiotic spindle and the different layers of the zona pellucida in human oocytes on the basis of birefringence in a non-destructive way. Conflicting results have been reported regarding the predictive value in ICSI cycles. OBJECTIVE: To assess the predictive ability of meiotic spindle and zona pellucida of human oocytes to implant by polarized microscopy in ICSI cycles. MATERIAL AND METHODS: Prospective and observational clinical study. 903 oocytes from 94 ICSI cycles were analyzed with polarized microscopy. Meiotic spindle visualization and zona pellucida birefringence values by polarized microscopy were correlated with ICSI cycles results. RESULTS: Meiotic spindle visualization and birefringence values of zona pellucida decreased in a direct basis with increasing age. In patients aged over the 35 years, the percentage of a visible spindle and mean zona pellucida birefringence was lower than in younger patients. Fertilization rate were higher in oocytes with visible meiotic spindle (81.3% vs. 64%; p < 0.0001), as well as embryo quality (47.4% vs. 39%; p=0.01). Fertilization rate was higher in oocytes with positive values of birefringence (77.5 % vs. 68.5% p=0.005) with similar embryo quality. Conception cycles showed oocytes with higher mean value of zona birefringence and visible spindle vs. no-conception cycles (p<0.05). CONCLUSIONS: Polarized light microscopy improves oocyte selection, which significantly impacts in the development of embryos with greater implantation potential. The use of polarized light microscopy with sperm selection methods, blastocyst culture and deferred embryo transfers will contribute to transfer fewer embryos without diminishing rates of live birth and single embryo transfer will be more feasible.
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Oócitos/citologia , Injeções de Esperma Intracitoplásmicas , Adulto , Birrefringência , Feminino , Humanos , Microscopia de Polarização , Prognóstico , Estudos Prospectivos , Fuso Acromático , Zona PelúcidaRESUMO
The S100A12 protein was validated as a biomarker of health status in porcine saliva samples using a semi-quantitative approach based on Western blotting in four healthy and sixteen diseased animals, and in four animals with severe respiratory disease during three days of antibiotic therapy. Afterwards, a non-competitive sandwich immunoassay was then developed, validated, and used to quantify S100A12 in clinical porcine samples, using 14 healthy and 25 diseased pigs. Finally, the S100A12 concentrations in the saliva of ten pigs with respiratory disease were monitored during antibiotic therapy. Diseased animals showed higher concentrations of S100A12 than healthy animals, and the high concentrations of S100A12 in pigs with respiratory distress were reduced after antimicrobial therapy. The assay developed showed good precision and accuracy, as well as a low limit of detection of 3.19 ng/mL. It was possible to store saliva samples at -20 °C, or even at 4 °C, for two weeks before analysis without losing the validity of the results. The concentrations of S100A12 observed in serum and saliva samples showed a moderately positive association with a correlation coefficient of 0.48. The concentrations of the new validated biomarker S100A12 are highly associated with the novel salivary biomarker of inflammation, adenosine deaminase, and moderately to highly associated with the total oxidant status. The results reported in this study provide a new way of evaluating inflammatory diseases in pigs using saliva samples, which should be further explored for disease prevention and monitoring in the field.
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Doenças Respiratórias , Doenças dos Suínos , Suínos , Animais , Proteína S100A12/análise , Proteína S100A12/metabolismo , Saliva/química , Biomarcadores/análise , Antibacterianos/metabolismo , Doenças Respiratórias/veterinária , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/metabolismoRESUMO
BACKGROUND: The concentration of biomarkers in saliva could be influenced by several factors not related to the specific condition under analyses, which should be considered for proper clinical interpretation. In the present study, the circadian rhythm of C-reactive protein (CRP), haptoglobin (Hp), Pig-MAP, S100A12, Cu, Zn, Adenosine deaminase (ADA), total protein (TP), total antioxidant capacity (TAC), total oxidant status (TOS), oxidative stress index (OSI), cortisol and α-amylase in saliva of 20 female and 20 male pigs was investigated. Moreover, the influence of sex and production phase (post-weaning, fattening and finishing) on the concentrations of biomarkers in a total of 414 healthy pigs was studied and the reference intervals for all salivary biomarkers were calculated accordingly. RESULTS: All parameters except Pig-MAP, OSI and α-amylase varied significantly along the daytime, and most of them peak around early afternoon (13-15 h). The cosinor analysis described the temporal dynamics of circadian rhythms for all parameters. The range values showed differences between male and female pigs in 8 out of the 13 biomarkers, with higher concentrations in females in comparison to male pigs. The influence of the production phase on the salivary concentrations was observed for all the biomarkers. The highest concentrations were observed for Pig-MAP, S100A12 and α-amylase in post-weaning animals, for TP in growing pigs and for OSI in finishing animals. Most of the sex-influenced biomarkers showed the highest concentrations at growing stages with some exceptions such as ADA or Hp that showed the peak at finishing and post-weaning stages respectively. CONCLUSIONS: It is necessary to establish the optimal daytime for routine saliva sampling to avoid circadian variations and for that end, the time interval between 10:00 a.m. to 12:00 a.m. is highly recommended. The factors sex and production phase influence the concentration of biomarkers and should be considered for proper biomarker interpretation. The reference intervals presented here for each salivary biomarker will help to correctly interpret the results of these analytes and contribute to the use of saliva as a non-invasive sample for the diagnosis and monitoring of the health status of swine farms.
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We present an optimized design for a 10G- differential-phase-shift-keyed (DPSK) receiver based on a silicon-on-insulator (SOI) unbalanced tunable Mach-Zehnder interferometer (MZI) switch in sequence with a Mach-Zehnder delay interferometer (MZDI). The proposed design eliminates the limitation in sensitivity of the device produced by the waveguide propagation losses in the delay line. A 2.3 dB increase in receiver sensitivity at a bit-error-rate (BER) of 10(-9) is experimentally measured over a standard implementation. The enhanced sensitivity is achieved with zero power consumption by tuning the operating wavelength or with less than 5 mW for a fixed wavelength using microheaters. Also the foot-print of the device is minimized to 0.11 mm(2) by the use of compact spirals.
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Interferometria/instrumentação , Processamento de Sinais Assistido por Computador/instrumentação , Telecomunicações/instrumentação , Desenho Assistido por Computador , Desenho de Equipamento , Análise de Falha de Equipamento , Sensibilidade e EspecificidadeRESUMO
We describe and demonstrate experimentally a method for photonic mixing of microwave signals by using a silicon electro-optical Mach-Zehnder modulator enhanced via slow-light propagation. Slow light with a group index of ~11, achieved in a one-dimensional periodic structure, is exploited to improve the upconversion performance of an input frequency signal from 1 to 10.25 GHz. A minimum transmission point is used to successfully demonstrate the upconversion with very low conversion losses of ~7 dB and excellent quality of the received I/Q modulated QPSK signal with an optimum EVM of ~8%.
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This paper analyzes the association between stress and immune response activations in different diseases, based on the salivary analytics. Moreover, a first attempt to discriminate between diseases was performed by principal component analysis. The salivary analytics consisted of the measurement of psychosocial stress (cortisol and salivary alpha-amylase) indicators, innate (acute phase proteins: C-reactive protein and haptoglobin), and adaptive immune (adenosine deaminase, Cu and Zn) markers and oxidative stress parameters (antioxidant capacity and oxidative status). A total of 107 commercial growing pigs in the field were divided into six groups according to the signs of disease after proper veterinary clinical examination, especially, healthy pigs, pigs with rectal prolapse, tail-biting lesions, diarrhea, lameness, or dyspnea. Associations between stress and immune markers were observed with different intensities. High associations (r = 0.61) were observed between oxidative stress markers and adaptive immune markers. On the other hand, moderate associations (r = 0.31-0.48) between psychosocial stress markers with both innate and adaptive immune markers were observed. All pathological conditions showed statistically significant differences in at least 4 out of the 11 salivary markers studied, with no individual marker dysregulated in all the diseases. Moreover, each disease condition showed differences in the degree of activation of the analyzed systems which could be used to create different salivary profiles. A total of two dimensions were selected through the principal component analysis to explain the 48.3% of the variance of our data. Lameness and rectal prolapse were the two pathological conditions most distant from the healthy condition followed by dyspnea. Tail-biting lesions and diarrhea were also far from the other diseases but near to healthy animals. There is still room for improvements, but these preliminary results displayed a great potential for disease detection and characterization using salivary biomarkers profiling in the near future.
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OBJECTIVE: The present study explores the possible factors related to severe cases of pandemic flu. DESIGN: A retrospective cohort study was conducted in patients hospitalized with Influenza A/H1N1 2009 during the pandemic period. SETTING: Ramon y Cajal University Hospital (Madrid, Spain). PATIENTS: All hospitalized patients with positive RT-PCR (real-time polymerase chain reaction) for Influenza A/H1N1 2009 virus. MAIN VARIABLES: The main variables collected were: history of risk factors for severe Influenza, history of immunization, clinical presentation, laboratory tests, chest X-ray report, administration of antiviral treatment, and hospital stay. RESULTS: The median age of the 100 cases was 38 years (range 4 months to 80 years). Seventy-seven percent of the patients had at least one risk factor. Asthma was the most common factor among patients younger than 18 years, versus smoking in the older subjects. Antiviral therapy was initiated a median time of three days (range 0 to 18 days) after the onset of illness. Nineteen percent of the patients were admitted to Intensive Care, and 2% died. Metabolic disease and abnormal chest X-ray findings were factors associated to admission to the ICU. CONCLUSION: As in other studies, abnormal chest X-ray findings upon admission and metabolic disease were related to poor outcomes of 2009 pandemic Influenza A (H1N1) infection in our patients.
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Vírus da Influenza A Subtipo H1N1 , Influenza Humana/epidemiologia , Unidades de Terapia Intensiva , Pandemias , Admissão do Paciente/estatística & dados numéricos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Criança , Pré-Escolar , Estudos de Coortes , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores de Risco , Espanha/epidemiologia , Adulto JovemRESUMO
This study evaluates if biomarkers of porcine health status in saliva samples is a mere reflection of serum to detect disease in pigs under field conditions. Four farms from the same commercial company were included to obtain samples from animals with different pathological conditions. A total of 10 healthy animals and 10-15 animals from each farm with clinical symptoms of the disease were sampled for paired saliva and blood during a veterinary clinical visit. The biomarker panel included acute-phase proteins (APPs), C-reactive protein (CRP), haptoglobin (Hp), an inflammatory marker, adenosine deaminase (ADA), the total antioxidant capacity (TAC), the levels of essential trace elements, copper (Cu) and zinc (Zn), and the measurement of the total protein content (TP). After detailed statistical analysis, the results showed that saliva could replace serum for APP measurements since a good agreement has been observed between the concentrations of APPs in both body fluids. For any other biomarker, no agreement between the concentrations quantified in serum and saliva samples was observed visually. However, salivary ADA and TP concentrations were statistically significantly higher in the diseased, whereas the statistical tests with serum concentrations were inconclusive. Furthermore, greater differentiation between healthy and diseased animals could be observed when the distribution of biomarkers was analysed in saliva than in other serum samples. The diagnostic power to discriminate between healthy and diseased pigs is similar in saliva and in serum samples. Preliminary regression models may offer an optimal combination of biomarkers for disease detection in saliva (Hp, CRP, and TAC) and serum (Hp, CRP, and Cu), which demands less labour, sample, and financial cost for saliva determinations. The contradictory results observed for TAC, Cu, and Zn levels between body fluids indicate a need for further studies. To sum up, saliva-based biomarkers instead of serum-based biomarkers could contribute to more efficient detection of diseased animals.
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Proteínas de Fase Aguda , Saliva , Animais , Biomarcadores , Proteína C-Reativa , Haptoglobinas , SuínosRESUMO
OBJECTIVE: To analyse compliance with the recommendations on the insertion-maintenance of peripheral venous catheter (PVC) and the incidence of complications according to the healthcare department that inserted the PVCs. PATIENTS AND METHODS: We conducted a prospective cohort follow-up study of PVCs, from their insertion in the emergency or internal medicine (IM) department until their withdrawal. RESULTS: We monitored 590 PVCs, 274 from the emergency department and 316 from IM. In terms of compliance with the process indicators, there was a cannulation rate in the antecubital fossa of 3.5 and 1.6 per 100 catheters-day (pâ¯<â¯.001) in the emergency and IM departments, respectively. The sterile placement rates were 1.6 and 12.4 (pâ¯<â¯.001), and the rate for transparent dressing was 2.1 and 11.5 (pâ¯<â¯.001) per 100 catheters-day in the emergency and IM departments, respectively. The complications rates showed no differences between the departments. The most common complication was phlebitis (95 cases, 16.1%). CONCLUSIONS: Compliance with the insertion-maintenance recommendations for PVC showed differences between the departments; however, the incidence of complications was similar.
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Cateterismo Periférico , Cateterismo Periférico/efeitos adversos , Catéteres , Seguimentos , Humanos , Medicina Interna , Estudos ProspectivosRESUMO
The aim of this investigation was to study the connection between body size, fatty acid composition and sensitivity to lipid peroxidation of heart mitochondria and microsomes isolated from different size bird species: manon (Lonchura striata), quail (Coturnix coturnix var japonica), pigeon (Columba livia), duck (Cairina moschata) and goose (Anser anser), representing a 372-fold range of body mass. Fatty acids of total lipids were determined using gas chromatography and lipid peroxidation was evaluated with a chemiluminescence assay. The fatty acids present in heart organelles of the different bird species analyzed showed a small number of significant allometric trends. In mitochondria, from the individual fatty acid data, palmitoleic acid (C16:1 n7) increased allometrically (r=0.878), while stearic acid (C18:0) was negatively related to body mass (r=-0.903). Interestingly, none of the calculated fatty acid variables, the average fatty acid saturated, monounsaturated, polyunsaturated (PUFA) and the unsaturation index (UI) was established to show significant body size-related variations. In heart microsomes, the content of C18:0 was significantly smaller (r=-0.970) in the birds of greater size. A significant allometric increase in linoleic acid (C18:2 n6) (r=0.986), polyunsaturated (r=0.990) and UI (r=0.904) was observed in the larger birds. The total n6 fatty acids of heart mitochondria did not show significant differences when it was correlated to body mass of the birds. Moreover, positive allometric relationships were shown for microsomes. The total n3 fatty acids of heart mitochondria and microsomes indicated no significant correlations to body mass of birds. The C16:1 n7, C18:0 in mitochondria and C18:0, C18:2 n6, PUFA, UI and PUFA n6 in microsomes showed significant differences when they were correlated to maximum life span (MLSP) of birds. As light emission=chemiluminescence originated from heart organelles was not statistically significant, a lack of correlation between the sensitivity to lipid peroxidation and body size or maximum life span was obtained. These results indicate that the high resistance of bird hearts to the attack by free radicals is body size-independent and would be related to the preservation of cardiac function.
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Aves/fisiologia , Peroxidação de Lipídeos , Microssomos/metabolismo , Mitocôndrias Cardíacas/metabolismo , Animais , Tamanho Corporal , Columbidae/fisiologia , Coturnix/fisiologia , Patos/fisiologia , Ácidos Graxos/metabolismo , Feminino , Gansos/fisiologia , Coração/anatomia & histologia , Longevidade , Masculino , Miocárdio/metabolismoRESUMO
Study objectives were to investigate whether C-reactive protein (CRP) in pig saliva could be quantified using an adapted, time-resolved immunofluorometry assay (TR-IFMA), and to determine whether the assay could distinguish healthy from diseased animals. The test method had intra- and inter-assay coefficients of variation of 5.75% and 9.73%, respectively, the limit of detection was 0.47ng/mL and the coefficient of determination was 0.98. Analysis of CRP concentrations in paired serum and saliva samples from 50 pigs gave a positive correlation (r=0.702, P<0.01) and the salivary CRP concentration was able to distinguish healthy from diseased animals in 62 samples from pigs with naturally occurring or experimentally-induced inflammation. The results suggest that this minimally invasive, straightforward and sensitive assay may be useful in pig health and welfare monitoring.
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Proteína C-Reativa/análise , Saliva/química , Doenças dos Suínos/diagnóstico , Animais , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Suínos , Fatores de TempoRESUMO
Hydatidosis is a zoonosis produced by the metacestode Echinococcus spp. The aims of this research are: to contribute to the knowledge of pediatric hydatidosis in the south-east of Buenos Aires province, to study its evolution from 1993 to 2002 at the Regional Maternity and Pediatric Hospital "Dr. Victorio Tetamanti", to determine the strains involved and to discuss the importance of the disease. The clinical records of diagnosed and/or operated patients were reviewed with regard to the hydatid disease. The strain was determined by using PCRs with Eg1 121a/122a primers. Forty-four cases were analyzed. Fifty nine point one per cent of the patients were boys. The mean age was 8 SD=3.8 years. Sixty one point four per cent had urban residence. Ultrasonography was used in 61% of the cases. The hepatic location was most frequently seen and the liver/lung ratio was 1.25. Ninety point nine per cent of patients received surgical treatment. Albendazole was used in 52% of cases. The average hospitalization time was 11 days. The G1/G2 strain group was determined. This report is the first one of its kind in the studied region. The permanence of hydatidosis in the region depends on the natural transmission of the parasite in the absence of control and prevention measures. The health authorities should implement strategies of prevention and control in the study area.
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Equinococose/epidemiologia , Adolescente , Animais , Anti-Helmínticos/uso terapêutico , Argentina/epidemiologia , Criança , Pré-Escolar , Terapia Combinada , Equinococose/tratamento farmacológico , Equinococose/parasitologia , Equinococose/cirurgia , Echinococcus granulosus/isolamento & purificação , Doenças Endêmicas , Feminino , Maternidades/estatística & dados numéricos , Hospitais Pediátricos/estatística & dados numéricos , Humanos , Lactente , Tempo de Internação/estatística & dados numéricos , Masculino , Estudos Retrospectivos , População Rural/estatística & dados numéricos , População Urbana/estatística & dados numéricosRESUMO
A time-resolved immunofluorometric assay was evaluated for measurement of C-reactive protein in meat juice from diaphragmatic muscle collected from slaughtered pigs. Analytical and clinical validation of the method was performed by using meat juice samples, obtained by freezing and thawing muscle pieces. The intra- and inter-assay coefficients of variation ranged from 2.2-5.8% to 7.9-14.3%, respectively. The limit of detection was 0.00038 microg/ml. The method measured the CRP concentrations in a linear manner with a good accuracy (r=0.99). CRP concentrations in serum were highly correlated with those in diaphragmatic meat juice (r=0.90; p<0.001). CRP concentrations were significantly higher in clinically affected pigs compared to non-diseased pigs. The assay described here provides a sensitive method for measuring CRP concentrations in meat juice, which can represent a suitable alternative to serum or blood samples and simplifies the process of sampling collection at slaughter.
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Proteína C-Reativa/química , Carne/análise , Animais , Líquidos Corporais/química , Proteína C-Reativa/metabolismo , Diafragma/química , Inflamação/metabolismo , Músculo Esquelético/química , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Suínos , Doenças dos Suínos/metabolismoRESUMO
The objective of this investigation was to examine the relationship between body size, fatty acid composition and sensitivity to lipid peroxidation of mitochondria and microsomes isolated from the brain of different size bird species: manon, quail, pigeon, duck and goose, representing a 372-fold range of body mass. Fatty acids of total lipids were determined using gas chromatography and lipid peroxidation was evaluated using a chemiluminescence assay. The allometric study of the fatty acids present in brain mitochondria and microsomes of the different bird species showed a small number of significant allometric trends. In mitochondria the percentage of monounsaturated fatty acids, was significantly lower in the larger birds (r=-0.965; P<0.008). The significant allometric increase in 18:2 n-6; linoleic acid (r=0.986; P<0.0143), polyunsaturated (r=0.993; P<0.007) and total unsaturated (r=0.966; P<0.034) in brain microsomes but not in mitochondria may indicate a preferential incorporation of this fatty acid in the brain endoplasmic reticulum of the larger bird species. The brain of all birds studied had a high content of docosahexaenoic acid. However brain mitochondria but not microsomes isolated from all the birds analyzed showed a significant decrease of arachidonic and docosahexaenoic acids during lipid peroxidation. The allometric analyses of chemiluminescence were not statistically significant. In conclusion our results show absence of correlation between the sensitivity to lipid peroxidation of brain mitochondria and microsomes with body size and maximum life span.
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Aves/metabolismo , Encéfalo/metabolismo , Ácidos Graxos/metabolismo , Peroxidação de Lipídeos , Microssomos/metabolismo , Mitocôndrias/metabolismo , Animais , Tamanho Corporal , Ácidos Graxos/química , Ácidos Graxos/isolamento & purificação , Luminescência , Microssomos/química , Mitocôndrias/química , Tamanho do Órgão , Especificidade da EspécieRESUMO
This study is a comparison of the efficiency of three technologies used for Y chromosome capture and the next-generation sequencing (NGS) technologies applied for determining its whole sequence. Our main findings disclose that streptavidin-biotin magnetic particle-based capture methodology offers better and a deeper sequence coverage for Y chromosome capture, compared to chromosome sorting and microdissection procedures. Moreover, this methodology is less time consuming and the most selective for capturing only Y chromosomal material, in contrast with other methodologies that result in considerable background material from other, non-targeted chromosomes. NGS results compared between two platforms, NextSeq 500 and SOLID 5500xl, produce the same coverage results. This is the first study to explore a methodological comparison of Y chromosome capture and genetic analysis. Our results indicate an improved strategy for Y chromosome research with applications in several scientific fields where this chromosome plays an important role, such as forensics, medical sciences, molecular anthropology and cancer sciences.
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Cromossomos Humanos Y/genética , Citometria de Fluxo/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Microdissecção e Captura a Laser/métodos , Análise de Sequência de DNA/métodos , Células Cultivadas , Cromossomos Humanos Y/química , Humanos , MasculinoRESUMO
Salivary glycoprotein profiles, obtained after boronic acid enrichment, were studied for the first time in pigs in order to search for specific overall alterations related to acute inflammatory condition. Five healthy pigs and five pigs suffering from rectal prolapse were used, and the levels of acute phase proteins were measured to determine the degree of inflammation of the animals. The enriched glycoprotein profiles, achieved by two-dimensional gel electrophoresis (2DE) were statistically evaluated and spots that appeared differentially regulated between states were subjected to MS analysis for protein identification. Spots from three unique proteins were identified: carbonic anhydrase VI (CA VI), α-1-antichymotrypsin and haptoglobin (Hp). CA VI appeared as two adjacent horizontal spot trains in the glycoprotein profile of healthy animals in its regular isoelectric points (pI). One spot of α-1-antichymotrypsin was found in saliva from pigs with rectal prolapse in an unusual basic pI, and was considered as a breakdown product. Hp was identified as several spot trains in saliva from pigs with rectal prolapse in an unusual alkaline pI and was consequently further investigated. SDS-PAGE and 2DE of paired serum and saliva samples combined with Western blot analysis showed that the unusual Hp position observed in saliva samples was absent in serum. Furthermore, N-glycans from serum and saliva Hp glycopatterns were evaluated from SDS-PAGE Hp bands and showed that the serum N-glycan distribution in Hp ß-chain was comparable in quantity and quality in both groups of animals. In saliva, no Hp ß-chain derived N-glycans could unambiguously be identified from this sample set, thus needing further detailed investigations in the future.
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Ácidos Borônicos/química , Haptoglobinas/metabolismo , Prolapso Retal/veterinária , Proteínas e Peptídeos Salivares/metabolismo , Doenças dos Suínos/diagnóstico , Animais , Western Blotting/veterinária , Eletroforese em Gel Bidimensional/veterinária , Eletroforese em Gel de Poliacrilamida/veterinária , Masculino , Espectrometria de Massas/veterinária , Prolapso Retal/diagnóstico , Prolapso Retal/etiologia , Saliva/química , Suínos , Doenças dos Suínos/etiologiaRESUMO
Polymerase chain reaction of a pentanucleotide microsatellite in the U1 snRNA gene complex generated a multiple band pattern due to the priming of paralogous sequences. Denaturation and slow renaturation of polymerase chain reaction products allow the formation of heteroduplex DNA that can be detected by its differential mobility in polyacrylamide gel electrophoresis. Heteroduplex analysis was used to determine if the U1 snRNA microsatellite could be a useful genetic marker in Echinococcus granulosus. A U1 snRNA microsatellite fragment from E. granulosus was isolated and characterized by Southern blot and sequencing. Four E. granulosus strains were analyzed: sheep, Tasmanian sheep, cattle, and camel strains. The former two showed polymorphism and shared three of the six patterns found for sheep strain. The cattle strain displayed two patterns, and the camel strain was monomorphic. The electrophoretic profiles were used for statistical analysis in order to determine genetic distance and the relationship among strains. Heteroduplex analysis can be helpful in genotyping E. granulosus strains and is useful in detecting polymorphism within strains.
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Echinococcus granulosus/genética , Repetições de Microssatélites/genética , Polimorfismo Genético/genética , RNA Nuclear Pequeno/genética , Animais , Sequência de Bases , Southern Blotting , Camelus , Bovinos , Eletroforese em Gel de Poliacrilamida , Marcadores Genéticos , Análise Heteroduplex , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , OvinosRESUMO
The aim of this work was to evaluate whether acute phase protein (APP) determinations could assist Official Veterinarians carrying out work in slaughterhouses. To test this hypothesis, the diagnostic accuracy of APP determinations in meat juice of pigs was analysed to differentiate between healthy and diseased pigs. One hundred and one pigs of two different origins were classified into two groups according to their health status (healthy and diseased pigs), which was determined by a veterinary clinical examination on the farm. To assess the pigs' immune status, against the main porcine diseases, serological analyses were monitored. A general idea of the degree of disease coverage was analysed by examining organ lesions postmortem. Haptoglobin (Hp) and C reactive protein (CRP) were measured in meat juice samples. 72.13 per cent of pigs appeared to be seropositive for the porcine respiratory and reproductive syndrome virus, and almost 86.2 per cent of them had concomitant infections with other pathogens, such as Porcine circovirus type 2 or Swine influenza virus. Median Hp and CRP concentrations were significantly higher in diseased animals at different stages of the production chain, when compared with levels found in healthy finishing pigs (P<0.0001). Receiver operating characteristic analysis showed the highest sensitivity-specificity pairs, nearly 80-90 per cent, at cut-off levels of 83 and 10 µg/ml for Hp and CRP determinations, respectively, with high AUCs 0.9. This cut-off could be useful for veterinary inspections at the time of slaughter, to differentiate between the carcase of a healthy animal and the carcase of an animal suffering from a systemic disease, which should be completely condemned.