Influence of the circadian cycle, sex and production stage on the reference values of parameters related to stress and pathology in porcine saliva.

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.

The Connection Between Stress and Immune Status in Pigs: A First Salivary Analytical Panel for Disease Differentiation.

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.

Transgenesis in parasitic helminths: a brief history and prospects for the future.

Helminth infections impact the health of hundreds of millions of persons globally and also cause important economic losses in livestock farming. Methodological limitations as well as the low attention given to the study of helminths have impacted biological research and, thus, the procurement of accurate diagnosis and effective treatments. Understanding the biology of helminths using genomic and proteomic approaches could contribute to advances in understanding host-helminth interactions and lead to new vaccines, drugs and diagnostics. Despite the significant advances in genomics in the last decade, the lack of methodological adaptation of current transgenesis techniques has hampered the progression of post-genomic research in helminthology. However, the application of new techniques, such as CRISPR, to the study of trematodes and nematodes has opened new avenues for genome editing-powered functional genomics for these pathogens. This review summarises the historical advances in functional genomics in parasitic helminths and highlights pending limitations that will need to be overcome to deploy transgenesis tools.

Echinostoma caproni (Trematoda): differential in vivo mucin expression and glycosylation in high- and low-compatible hosts.

Enhanced mucus production and release appears to be a common mechanism for the clearance of intestinal helminths, and this expulsion is normally mediated by Th2-type immune responses. To investigate the factors determining the expulsion of intestinal helminths, we have analysed in vivo expression of mucin genes at the site of infection in two host species displaying different compatibility with Echinostoma caproni (Trematoda). Surprisingly, a general down-regulation on mucin mRNA expression was detected in low-compatible hosts (rats) coinciding with the development of Th2/Th17 responses and the early rejection of the worms from the intestinal lumen. This suggests the existence of a mechanism by which the parasites can modulate the mucus barrier to favour their survival. In highly compatible hosts (mice), some mucin genes were found to be up-regulated throughout the infection, probably, to protect the intestinal epithelium against the infection-induced inflammation developed in this host species. Moreover, infection-induced changes on mucin glycans were also studied by lectin histochemistry. Similar alterations were detected in the ileum of infected mice and rats, except with SNA lectin, indicating that sylated mucins might play an important role in determining the evolution of the infection in each host species.

The use of banquettes of Posidonia oceanica as a source of fiber and minerals in ruminant nutrition. An observational study.

In the search for new food sources that contribute to the optimization of livestock production this paper discusses the possibility of using waste (called banquettes) of a marine plant commonly found on the Mediterranean coasts: Posidonia oceanica. The idea stems from the use of a waste that in summertime generates large costs because it is considered bothersome on the beaches. Thus, tons and tons of residues are collected each year from the beach, being destined for incineration. However, alternative uses for these residues are suggested, such as forage that is particularly relevant for the Mediterranean coast, where the weather does not support abundant grass growth. With this purpose, samples of banquettes of P. oceanica from six different points of a touristic place located in the Murcia Region (S.E. of Spain) were collected in April 2012 on the same day directly from the beach above the water line, washed with distilled water and sun-dried for 48 h. Approximately 500 g of each sample of plant material was chopped and two subsamples of 200 g each were placed in airtight plastic containers and sent to the laboratory for mineral and chemical analysis. This report provides data on the nutritional composition of P. oceanica such as mineral contents (As, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn) and chemical composition (ash, CP, ether extract (EE), NDF and ADF, respectively) and ADL. Finally, the in sacco rumen disappearance of dry matter (DM), organic matter (OM), CP and NDF were studied. Our results showed that minerals, except for Fe, where within the range of values reported for P. oceanica in other parts of the Mediterranean region. Given the high Fe content further studies assessing the antagonic Fe-Cu interaction and its effect on animal health should be addressed. In relation to chemical composition, it is clear that this seagrass is a poor protein source and has levels of DM degradability at 24 h, similar to those obtained for cereal straw. The information summarized here shows some potential for the use of banquettes of P. oceanica as forage source in ruminant nutrition.

Th17 responses in Echinostoma caproni infections in hosts of high and low compatibility.

In order to investigate the factors determining the expulsion of intestinal helminths, we have analyzed the in vivo expression of IL-17, TGF-ß and IL-23 in several tissues of two host species displaying different compatibility with Echinostoma caproni (Trematoda). We did not observe upregulation of these cytokines in any of the tissues of the high compatible host (mice). In contrast, the responses in the host of low compatibility (rats) with the parasite were markedly different. Significant increases in the expression of IL-17 and TGF-ß were observed in the Peyer's patches and the intestine from the 2 to 8 weeks post-infection. The expression of IL-23 was upregulated from 2 to 4 weeks post-infection in the spleen, Peyer's patches and the intestine. Considering together our results with those published previously the development of chronic infections appears to be related with the development of local Th1 responses, whereas the early rejection of the worms is mediated by the development a biased Th17/Th2 phenotype. The Th17 response generated in rats may facilitate the worm expulsion via the suppression of the inflammatory Th1 responses and the increase in intestinal contractility.

Proteomic analysis of Strongyloides stercoralis L3 larvae.

Strongyloidiasis can be perpetuated by autoinfection with the filariform larvae L3, causing asymptomatic chronic infections and creating a population of carriers, affecting not only developing countries. So far, very little is known about the proteins that interact with the human host, and few proteins from the infective Strongyloides stercoralis L3 have been characterized. Here, we report results obtained from a proteomic analysis of the proteins from S. stercoralis L3 larvae obtained from patients. Since the genome of S. stercoralis is not yet available, we used proteomic analysis to identify 26 different proteins, 13 of them released by short digestion with trypsin, which could represent surface-associated proteins. The present work extends our knowledge of host-parasite interactions by identifying proteins that could be of interest in the development of diagnostic tools, vaccines, or treatments for a neglected disease like strongyloidiasis.

Atrial flutter and myocardial infarction-like ECG changes as manifestations of left ventricle involvement from lung carcinoma

Lung cancer involvement of the heart is not unusual, but in most cases is silent. Arrhythmia and electrocardiographic findings suggesting an acute myocardial infarction could be the first manifestation of myocardial infiltration by the tumour. Echocardiography could be a valuable tool to define the diagnosis in patients with lung cancer and newly diagnosed arrhythmia or ST-T wave alterations. When echocardiographics findings are not conclusive, magnetic resonance imaging (MRI) allows differentiation between tumour and myocardium (AU)

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Identification of antigenic proteins from Echinostoma caproni (Trematoda) recognized by mouse immunoglobulins M, A and G using an immunoproteomic approach.

Antigenic proteins of Echinostoma caproni (Trematoda) against mouse IgM, IgA, IgG, IgG1 and IgG2a were investigated by immunoproteomics. Excretory/secretory products (ESP) of E. caproni separated by two-dimensional (2D) gel electrophoresis were transferred to nitrocellulose membranes and probed with the different mouse immunoglobulin classes. A total of four proteins (enolase, 70 kDa heat-shock protein (HSP-70), actin and aldolase) were accurately identified. Enolase was recognized in eight different spots of which seven of them were detected in the expected molecular weight and were recognized by IgA, IgG or IgG and IgG1. Another spot identified as enolase at 72 kDa was only recognized by IgM. Digestion with N-glycosidase F of the 72 kDa band rendered a polypeptide with an apparent molecular weight similar to that expected for enolase recognized by Western immunoblotting using anti-enolase antibodies. This suggests that glycosylated forms of enolase may be involved in the early thymus-independent responses against E. caproni. Early IgM responses were also generated by actin and the HSP-70 which suggests that these proteins are exposed early to the host and may be of importance in the parasite establishment. The IgA responses also appear to be mediated by the HSP-70 and aldolase which could be related with the close contact of these proteins with the host mucosal surface after secretion.

Disposition kinetics and pharmacokinetics--pharmacodynamic integration of difloxacin against Staphylococcus aureus isolates from rabbits.

The pharmacokinetics of difloxacin were studied following intravenous (IV), subcutaneous (SC) and oral administration of 5mg/kg to healthy white New Zealand rabbits (n = 6). Difloxacin concentrations were determined by HPLC assay with fluorescence detection. Minimal inhibitory concentrations (MICs) assay of difloxacin against different strains of S. aureus from different european countries was performed in order to compute the main pharmacodynamic surrogate markers. The plasma difloxacin clearance (Cl) for the IV route was (mean +/- SD) 0.41 +/- 0.05 L/h kg. The steady-state volume of distribution (V(ss)) was 1.95 +/- 0.17 L/kg. The terminal half-life [Formula: see text] was (mean+/-SD) 4.19+/-0.34 h, 7.53 +/- 1.32 h and 8.00 +/- 0.45 h after IV, IM and oral, respectively. From this data, it seems that a 5 mg/kg dose difloxacin would be effective by SC and oral routes in rabbits against bacterial isolates with MIC0.1 microg/mL.

Leucine aminopeptidase is an immunodominant antigen of Fasciola hepatica excretory and secretory products in human infections.

The liver fluke Fasciola hepatica parasitizes humans and ruminant livestock worldwide, and it is now being considered a reemerging zoonotic disease, especially in areas in which it is endemic, such as South America. This study investigates the immune response to excretory and secretory products produced by F. hepatica in a group of patients from the Peruvian Altiplano, where the disease is highly endemic. Using a proteomic approach and immunoblotting techniques, we have identified the enzymes leucine aminopeptidase (LAP) and phosphoenolpyruvate carboxykinase as immunodominant antigens recognized by sera from fasciolosis patients. An indirect enzyme-linked immunosorbent assay using recombinant LAP as the antigen was developed to check sera from individuals of this region. Our results demonstrate that LAP produces a specific and strong reaction, suggesting its potential use in the serologic diagnosis of F. hepatica infections in humans.

Plasma malonaldehyde (MDA) and total antioxidant status (TAS) during lactation in dairy cows.

Metabolic profiles are widely used to monitor health, reproductive status and nutritional status. In the last few years, the evaluation of oxidative stress has contributed increasingly to our knowledge of the fundamental mechanisms involved in metabolic disorders, especially important in dairy cows, in which lactation imposes great physiological demands on the body's homeostatic mechanisms. The aim of the present study was to evaluate oxidative status in healthy cows during lactation (from lactation onset to peak lactation) using two parameters: (i) plasma levels of malondialdehyde (MDA) and (ii) total antioxidant status (TAS). Our results confirm that nutrition can influence the characteristic metabolic changes occurring between lactation onset and peak lactation. In addition, the combination of MDA and TAS can provide complementary information about the metabolic status of the cow. Thus, the proper metabolic adaptation to the onset of lactation, showed by metabolic profiles, contrasts with the high levels of free radicals which cause lipid peroxidation and high MDA values (68.99+/-33.64 microm/L) which is maintained only for a short period of time. In this moment, the antioxidant system can cope efficiently with lipoperoxide production. The most remarkable fact was the great inter-individual variations observed in MDA that might be studied in further investigations. When the animal reaches peak lactation, metabolic status is stabilized, and this is reflected by antioxidant status with mean values of 28.87+/-5.33 microm/L for MDA and 0.154+/-0.002 mmol/L for TAS values.

[Right ventricular thrombosis and pulmonary thrombosis associated with venous-peritoneal diversion (Le Veen). Diagnosis with transesophageal echocardiography]. / Trombosis ventricular derecha y embolismo pulmonar asociados a derivación peritoneo-venosa (Le Veen). Diagnóstico por ecocardiografía transesofágica.

Central venous thrombosis (CVT) and pulmonary embolism (PE) are complications that have been reported in association with the use of venous-peritoneal shunts (Le Veen). CVT usually develops around the proximal end of the catheter; the clinical course is varied and usually requires venous imaging to confirm the diagnosis. We present a case of CVT associated with PE, in which the thrombus was located in the right ventricular cavity (distal to the catheter tip). Two-dimensional transesophageal echocardiography was used for diagnosis and follow-up.

[Clinical evaluation of the changes produced by nitrates in patients with chronic heart insufficiency]. / Valoración clínica de los cambios producidos por los nitratos en pacientes con insuficiencia cardíaca crónica.

The aim for this study was to evaluate the hemodynamic changes that appear in patients with myocardiopathy at functional stage II (NYHA) subjected to standard treatment (digital+diuretics) plus nitrates. We used isosorbide-5-mononitrate (IS-5-MN). We studied 20 patients for 100 days in different phases. The placebo-controlled, crossover, double-blind stage random distribution was from day 2 to 5. From day 6 to 20 all the patients took 40 mg/12 hours of IS-5-MN, taking 40 mg/8 hours of active ingredient from day 21 to 100. We performed basal cycloergometrics 3 and 6 hours after administration on days 2, 5, 20, 34 and 100 of the treatment, as well as echocardiograms on these same days. We compared the results obtained versus placebo (p < 0.05). There were no significant differences in heart rate at rest or systolic arterial tension at rest or maximum. Maximum heart rate increased significantly in the tests at days 34 (basal: 143 +/- 9; 3 hours: 139 +/- 8; 6 hours: 140 +/- 7) and 100 (basal: 145 +/- 7; 3 hours: 141 +/- 8; 6 hours: 142 +/- 6) versus the tests carried out with placebo (basal: 136 +/- 8; 3 hours: 135 +/- 7; 6 hours: 136 +/- 7). There were no differences in the tests after acute dose or at treatment day 20.(ABSTRACT TRUNCATED AT 250 WORDS)

[Cardiac rhabdomyosarcoma. Its echographic diagnosis]. / Rabdomiosarcoma cardíaco. Diagnóstico ecocardiográfico.

We report a case of cardiac rhabdomyosarcoma whose initial clinical features were fever and palpitations due to documented ventricular tachycardia. Sequential two-dimensional echocardiographic studies pointed out the presumed diagnosis, showing intracavitary masses at multiple sites appearing within a short period of time. The postmortem examination confirmed the diagnosis of rhabdomyosarcoma.

[Doppler echocardiography assessment of cardiac abnormalities in parenteral drug addicts]. / Valoración de las alteraciones cardíacas por ecocardiografía-Doppler de los pacientes adictos a drogas por vía parenteral.

We studied by Echocardiographic-Doppler 114 consecutive intravenous drugs addicts (IVDA); 91 were positive human immunodeficiency virus (HIV+) and 23 negatives. We classified them in five groups; beginning the negative HIV as group 0, and groups I to IV stratified according the Central Disease Control (CDC) classification. We compared the cardiac abnormalities founded between themselves and a control group presumed healthy persons of similar age. The cardiac cavities dimensions showed a statistic significant increased left ventricular end-systolic and diastolic diameters, right ventricular diameter, posterior wall and interventricular septum thickness and aortic root diameter compared with the control group; but all were in the normal range for age. The left ventricular fractional shortening was statistically different from control group related the other groups, and the group IV related other. The existence and severity of pericardial effusions were directly related to the illness stage. We founded moderate pericardial effusions in 25% patients in the 0 to III groups, increasing until 50% in the group IV. The presence of valvular vegetations, nearly 30% in our series, ought to the IVDA. We did not found relationship between the severity of valvular incompetence and the illness stage. We recorded a excellent correlation between the ratio T4/T8 lymphocytes with the progress of illness and the existence and severity of cardiac abnormalities.

[Cor triatriatum in the adult. Its diagnosis by Doppler echocardiography]. / Cor triatriatum en el adulto. Diagnóstico por ecocardiografía-Doppler.

We present an adult with echocardiographic diagnosis of cor triatriatum. Continuous wave Doppler echocardiography was able to assess correctly the severity of the left ventricular inflow obstruction caused by the intra-atrial membrane. Even though the diagnosis was confirmed by catheterization, the surgical decision was based in noninvasive data. Postoperative Doppler echocardiography proved the disappearance of the intra-atrial obstruction. The anatomical information obtained by echocardiography as well as the assessment of the intra-atrial obstruction by cardiac Doppler seem to be sufficient to make surgical decisions in patients suffering from this disease. Both are also suitable to test the result of the surgical intervention.

Acute myocardial infarction after blunt chest trauma in young people: need for prompt intervention.

We describe four cases of acute myocardial infarction in young patients, secondary to blunt chest trauma. One case was treated with intracoronary thrombolysis and angioplasty, two cases received systemic thrombolysis, and the last one did not have any reperfusion therapy. The coronary angiograms of the left anterior descending artery showed thrombosis in two cases, coronary dissection in one case, and no morphological lesions in the other. We encourage the early performance of angiographic studies in these patients, adjusting the therapy to their pathophysiologic mechanism.

Two-dimensional echocardiographic features of echinococcosis of the heart and great blood vessels. Clinical and surgical implications.

Echocardiographic findings in cardiac echinococcosis have been previously described in isolated cases, but no detailed account of the various echocardiographic aspects and their clinical and surgical implications have ever been reported. We present a retrospective analysis of the two-dimensional echocardiograms of 15 patients with hydatid cysts affecting the heart or great blood vessels. Two-dimensional echocardiogram features were compared with the main clinical syndromes. Surgical confirmation of the echocardiogram findings was available in 12 patients. The cyst sizes ranged from 0.5 to 12 cm in diameter. Eleven patients had single cysts, three patients had two cysts, and one patient had multiple cysts. Cysts were located in the intramyocardial region in nine patients, the pericardial in three, and the paracardial in another three. All intramyocardial cysts protruded into the adjacent cardiac chamber, but in only two patients was there significant tricuspid valvular dysfunction. Pericardial and mediastinal cysts showed compression of cardiac chambers or great blood vessels, and two cysts had ruptured into the descendent thoracic aorta or inferior vena cava. In most patients, two-dimensional echocardiographic images of hydatid cysts were those of a cystic mass having well-defined edges and internal trabeculations corresponding to daughter membranes. However, in four patients, two-dimensional echocardiographic images showed a "solid" mass instead of a cystic mass, and in one patient with the multivesicular variety of echinococcosis, the images showed a large mass with poorly defined edges having a honeycombed appearance causing lysis of the anterior arch of the second and third left ribs. The two-dimensional echocardiographic finding showed good correlation with main clinical syndromes, but anaphylactic reactions occurred in every cyst location. In one patient, postoperative two-dimensional echocardiography showed two small intramyocardial cysts that had not been noticed during preoperative two-dimensional echocardiography or during surgical examination. Pathological examination in the four patients with a solid mass showed replacement of the hydatid liquid by necrotic matter containing membrane residues with a foreign-body inflammatory reaction of a granulomatous type. In conclusion, two-dimensional echocardiography is a very useful tool for diagnosis and management of patients with cardiac echinococcosis, but the great diversity of findings regarding number, size, location, and appearance of cysts must be borne in mind to interpret correctly the two-dimensional echocardiograms.