Basics for the potential use of saliva to evaluate stress, inflammation, immune system, and redox homeostasis in pigs.

The use of saliva as a biological sample has many advantages, being especially relevant in pigs where the blood collection is highly stressful and painful, both for the animal and the staff in charge of the sampling. Currently one of the main uses of saliva is for diagnosis and detection of infectious diseases, but the saliva can also be used to measure biomarkers that can provide information of stress, inflammation, immune response and redox homeostasis. This review will be focused on the analytes that can be used for such evaluations. Emphasis will be given in providing data of practical use about their physiological basis, how they can be measured, and their interpretation. In addition, some general rules regarding sampling and saliva storage are provided and the concept of sialochemistry will be addressed. There is still a need for more data and knowledge for most of these biomarkers to optimize their use, application, and interpretation. However, this review provides updated data to illustrate that besides the detection of pathogens in saliva, additional interesting applicative information regarding pigs´ welfare and health can be obtained from this fluid. Information that can potentially be applied to other animal species as well as to humans.

Insulin in the saliva of pigs: Validation of an automated assay and changes at different physiological conditions.

This study aimed to evaluate whether insulin could be measured in the saliva of pigs and if its concentration changes in some physiological conditions. For this purpose, a validation of an automated heterologous immunoassay for measuring insulin in the saliva of pigs was performed. In addition, the possible changes of salivary insulin concentration in sows after food intake and during gestation and lactation were studied. The evaluated immunoassay was able to detect insulin in the saliva of pigs in a precise and accurate way when species-specific calibrators were used. There was no correlation in insulin concentrations between serum and saliva. Insulin concentrations showed a significant increase in the saliva of sows after feeding. Sows at farrowing and lactation presented higher salivary insulin levels as compared with those in gestation. In conclusion, the results showed that insulin could be measured in the saliva of pigs, and changes in its concentration can be detected due to food intake and different physiological conditions.

Changes in saliva analytes during pregnancy, farrowing and lactation in sows: A sialochemistry approach.

Salivary biomarkers were studied in 17 healthy Large White sows from early gestation to the end of lactation. Saliva samples were obtained at 34 ± 3 days from insemination (G30), 24 ± 4 days before farrowing (G90), within the first 24 h after farrowing (L1) and at the end of a lactation period of 21 days (L21). The measurements in saliva included stress-related biomarkers (cortisol, chromogranin A, α-amylase, butyrylcholinesterase [BChE] and lipase [Lip]), inflammatory biomarkers (adenosine deaminase isoenzymes 1 [ADA1] and 2 [ADA2], and haptoglobin [Hp]) and oxidative stress biomarkers (cupric reducing antioxidant capacity, trolox equivalent antioxidant capacity, ferric reducing ability, uric acid, advanced oxidation protein products [AOPP] and hydrogen peroxide [H2O2]), as well as routine biochemistry analytes (aspartate aminotransferase [AST], alkaline phosphatase [ALP], γ-glutamine transferase [GGT], lactate dehydrogenase [LDH], creatine kinase [CK], urea, creatinine, triglycerides, lactate, calcium and phosphorus). The main changes were observed at farrowing, with increases in biomarkers of stress (cortisol and BChE), inflammation (ADA isoenzymes and Hp) and oxidative stress (AOPP and H2O2), as well as muscle and hepatic enzymes (CK, AST, ALP, GGT and LDH). Lactate and triglycerides increased at the end of gestation and remained at high concentrations until the end of lactation. Lip was higher in gestation than at lactation. Thus, changes in biomarkers of stress, immune function, oxidative stress, hepatic and muscle integrity, and energy mobilization occur in sow saliva during pregnancy, farrowing and lactation. These changes, caused by physiological conditions, should be taken into consideration when these biomarkers are used for the evaluation of sow health and welfare.

Measurement of cortisol, cortisone and 11ß-hydroxysteroid dehydrogenase type 2 activity in hair of sows during different phases of the reproductive cycle.

Two sensitive assays based on AlphaLISA technology were developed and validated for the measurement of cortisol and cortisone in hair of pigs, that also enabled estimation of 11ß-hydroxysteroid dehydrogenase type 2 activity. These assays were applied to hair samples from sows (n = 32) collected at 5 days before, and at 23 and 59 after farrowing, in reproductive cycles in two different periods: spring-summer (n = 16) and winter-spring (n = 16). The assays were precise (imprecision <12%) and accurate (recovery range, 80-115%) for cortisol and cortisone determination. Hair cortisone concentrations and the cortisone/cortisol ratio (an estimate of 11ß-hydroxysteroid dehydrogenase isoenzyme type 2 activity) increased after farrowing more than cortisol, being these changes of higher magnitude during periods of higher atmospheric temperature. The measurement of hair cortisone concentrations and estimations of the activity of the 11ß-hydroxysteroid dehydrogenase isoenzyme type 2, measured by the assays developed in this study, are complementary biomarkers to hair cortisol, and can increase at periods associated with stress, such as farrowing and lactation, especially at high atmospheric temperatures. .

Homocysteine measurement in pig saliva, assay validation and changes after acute stress and experimental inflammation models: A pilot study.

High homocysteine (Hcy) concentration in serum has been associated to stress and inflammation in humans, but this association has not been studied in saliva in any animal species. The purpose of this research was to study salivary Hcy levels in pigs under stressful and inflammatory conditions. A commercially available enzyme-linked immunosorbent assay specific for Hcy determination in pigs was adapted and validated in saliva, yielding reproducible and accurate results. Hcy was measured in paired serum-saliva samples and no correlation was observed between serum and salivary Hcy. Salivary Hcy was measured in two experimental models of stress induction in pigs: restraint with a nasal snare and isolation. Homocysteine concentration and the homocysteine to total protein (Hcy/TP) ratio significantly increased 15min after restraining and decreased after some days of isolation. Significant correlation was observed between Hcy and chromogranin A. After an experimentally induced inflammation by subcutaneous turpentine injection, salivary Hcy increased only 3h after turpentine administration; however, the Hcy/TP ratio did not show any change. No correlation was found between salivary Hcy and serum C-reactive protein. In conclusion, salivary Hcy concentration increased when pigs were restrained with a nasal snare or stressed by isolation, probably reflecting an increase in the sympathetic activity. On the other hand, Hcy increased after an experimental inflammation induced by turpentine administration but in this case probably reflects an increase in total protein production in saliva.

Understanding and simulating the material behavior during multi-particle irradiations.

A number of studies have suggested that the irradiation behavior and damage processes occurring during sequential and simultaneous particle irradiations can significantly differ. Currently, there is no definite answer as to why and when such differences are seen. Additionally, the conventional multi-particle irradiation facilities cannot correctly reproduce the complex irradiation scenarios experienced in a number of environments like space and nuclear reactors. Therefore, a better understanding of multi-particle irradiation problems and possible alternatives are needed. This study shows ionization induced thermal spike and defect recovery during sequential and simultaneous ion irradiation of amorphous silica. The simultaneous irradiation scenario is shown to be equivalent to multiple small sequential irradiation scenarios containing latent damage formation and recovery mechanisms. The results highlight the absence of any new damage mechanism and time-space correlation between various damage events during simultaneous irradiation of amorphous silica. This offers a new and convenient way to simulate and understand complex multi-particle irradiation problems.

Evaluation of a cushioned method for centrifugation and processing for freezing boar semen.

The purpose of this investigation was to evaluate the use of an iodixanol cushion during centrifugation on sperm recovery and yield after centrifugation (sperm recovery, sperm motility, viability, membrane lipid disorder, acrosome reaction and ROS generation); and to investigate how this procedure affects sperm function after freezing-thawing (sperm motility, membrane lipid disorder, acrosomal status and homologous in vitro penetration test). The sperm-rich fractions from fertile boars were centrifuged under two centrifugation régimes: 800xg for 10min (standard method) and 1000xg for 20min with an iodixanol (60% w/v) cushion at the bottom of the centrifuge tubes (Cushion method). The highest recovery was achieved using the cushion method (sperm loss for cushion method was 0.50%+/-0.18 versus 2.97%+/-0.43 for standard method, P<0.01) and sperm quality was not significantly affected by the centrifugation régime. The motion parameters (% progressive motility, % motility, VCL, VSL, VAP, ALH, BCF, P<0.05) of frozen-thawed samples showed higher values using the standard method. However, a higher number of viable spermatozoa with lower lipid disorders were found in spermatozoa processed with the cushion method. The in vitro penetration assay showed that the individual boar influenced the parameters studied but there were no differences between the two centrifugation régimes used. Our results support the hypothesis that the proportion of sperm loss in frozen-thawed semen was significantly influenced by the centrifugation régime. Therefore, the iodixanol cushion method is a suitable tool for cryopreservation of boar semen in order to reduce sperm loss without affecting sperm quality.

Digital storage phosphor chest radiography: an ROC study of the effect of 2K versus 4K matrix size on observer performance.

PURPOSE: To compare observer performance in the detection of abnormalities on 1,760 x 2,140 matrix (2K) and 3,520 x 4,280 matrix (4K) digital storage phosphor chest radiographs. MATERIALS AND METHODS: One hundred sixty patients who underwent dedicated computed tomography (CT) of the thorax were prospectively recruited into the study. Posteroanterior and lateral computed radiographs of the chest were acquired in each patient and printed in 2K and 4K formats. Six radiologists independently analyzed the hard-copy images and scored the presence of parenchymal (opacities 2 cm, and subtle interstitial), mediastinal, and pleural abnormalities on a five-point confidence scale. With CT as the reference standard, observer performance tests were carried out by using receiver operating characteristic (ROC) analysis. RESULTS: Analysis of averaged observer performance showed 2K and 4K images were equally effective in detection of all three groups of abnormalities. In the detection of the three subtypes of parenchymal abnormalities, there were no significant differences in averaged performance between the 2K and 4K formats (area below ROC curve [A(z)] values: opacities 2 cm, 0.86 +/-.025 and 0.85 +/- 0.030; subtle interstitial abnormalities, 0.73 +/- 0.041 and 0.72 +/- 0.041). Averaged performance in detection of mediastinal and pleural abnormalities was equivalent (A(z) values: mediastinal, 0.70 +/- 0.046 and 0.73 +/- 0.033; pleural, 0.85 +/- 0.032 and 0.86 +/- 0.033). CONCLUSION: Observer performance in detection of parenchymal, mediastinal, and pleural abnormalities was not significantly different on 2K and 4K storage phosphor chest radiographs.

Primary bronchogenic carcinoma after heart or lung transplantation: radiologic and clinical findings.

Chronic immunosuppression in organ transplant recipients predisposes to the development of malignant disease. The authors describe their 29-year institutional experience of bronchogenic carcinoma developing after heart and lung transplantation. Seven cases of bronchogenic carcinoma were diagnosed in 1,119 heart and lung transplant recipients. Computed tomography scans and radiographs at time of diagnosis, as well as prior radiographs available in six patients were retrospectively analyzed by two radiologists in consensus. The seven cases involved six heart and one lung transplant recipients. Six patients were smokers with a mean smoking history of 66 pack-years. Mean time interval from transplantation to cancer detection was 25 months. Radiologic findings consisted of a solitary pulmonary nodule (n = 3), mass with satellite nodules (n = 1), and obstructive pneumonitis (n = 1). In the sixth patient, the cancer was not radiographically visible because of obscuration by adjacent fibrosis. On review, radiographic abnormalities were present a mean of 12 months prior to diagnosis in 66% of patients. In the heart or lung transplant population, bronchogenic carcinoma develops in recipients with extensive smoking histories. It presents radiographically as a nodule, mass, or obstructive pneumonitis, and is usually visible on radiographs before the time of diagnosis.

[Effect of a highly hypocaloric diet on the control of morbid obesity in the short and the long term]. / Efecto de una dieta altamente hipocalórica sobre el control de la obesidad mórbida a corto y medio plazo.

BACKGROUND: The ponderal evolution and biochemical parameters during the in patient administration of a highly hypocaloric and high protein diet (VLCD) were evaluated. The efficacy at medium term of this treatment was determined and the possible determining factors of the ponderal evolution were analysed. METHODS: A VLCD was administered to 20 women with morbid obesity during 21 days. The modifications of the biochemical parameters were determined during admission. The ponderal evolution was studied during hospitalization and during 9 months of out patient follow up. The prognostic factors of ponderal evolution analyzed were: age, attempts to lose weight, familiar history of obesity, maximum weight attained, weight and index of body mass (IBM) at study initiation and admission, loss of weight during admission and the number of out patient controls. RESULTS: During hospitalization, a mean loss of 7.0-1.9 kg, a significant decrease in glycemia, cholesterol, triglycerides, lymphocytes and blood pressure were observed. It was found that ponderal loss at 3 and 6 months may be predicted by IBM upon initiation of the study. At 9 months weight was lower than at initiation of hospitalization but not than at discharge (p < 0.001). Weight upon initiation of the study and the maximum reached explain the weight at 9 months in 82.5% of the cases. CONCLUSIONS: Highly hypocaloric and high protein diets lead to a significant weight loss which is maintained at medium term and an improvement in the factors of cardiovascular risk are observed. Of the factors evaluated, only the grade of obesity significantly influences ponderal evolution.

Clinical and histological observations on the application of intermingled auto- and porcine-skin heterografts in third degree burns.

Escharectomy and skin grafting with both homograft and porcine skin has become an effective method in treating massive third degree burns. Seventeen patients and 21 operations of intermingled transplantation of auto- and fresh porcine skin heterografts after escharectomy of the severe burn wounds have been carried out since March 1973. Clinical and histological data are summarized, among which we observed the 'fusing phenomena' of auto- and porcine skin heterografts in 6 patients. Vascularization, 'turning red', viability and rejection as well as ways to improve the results of the grafting method are discussed. No vascular communication between fresh porcine skin and the host wound has been observed during the early postoperative period. The cause of 'turning red' is a reddish transudation between the graft and the host wound seen through the thin porcine skin. Based on histological observations, porcine skin is viable after transplantation. With nutritional support apparently coming from the underlying plasma and tissue fluid. Eventually the process of rejection is similar to that of homograft Better results are found with porcine skin grafts 0.4-0.5 mm in thickness placed 0.5-0.75 cm apart. In order to avoid large sloughing wound surfaces less than 20 per cent area of porcine skin coverage is advisable.

Complications of laparoscopic sterilization. Comparison of 2 methods.

A retrospective study of 5346 cases of laparoscopic sterilization with special reference to early and late complications was undertaken. In the 6-year period from 1972 to 1978, 846 patients were sterilized by the elasticized silicone-ring technique and 4500 patients were sterilized by the electrocautery method. The electrocautery method was complicated by electrical burns in 13 cases (0.29%); 3 of these patients required bowel resections. Mechanical complications occurred in 1.6% of the Silastic-ring cases, but none of the patients required additional surgery. The Silastic-ring patients had greater postoperative abdominal pain than the electrocautery patients, but it usually abated within 48 hours. There were 15 cases of postoperative pelvic infection in the electrocautery group, and none in the Silastic-ring group. Bleeding from the mesosalpinx occurred in both groups and occurred more often with coincident suction D&C. Technical failure to perform the sterilization procedure was principally related to previous abdominal surgery. A substantial number of patients complained of menstrual irregularity and/or dysmenorrhea following these sterilization procedures. The majority of women reported unchanged or improved sexual relations. Improvement in sexual relations was reported by significantly more patients in the Silastic-ring group than in the electrocautery group.

PIP: 5346 cases of laparoscopic sterilization, using both electrocautery and Silastic-ring method, were studied from 1972-1978. Of the 4500 electrocautery method patients there were 13 cases of electrical burns, 3 of which required bowel resection. Of the 846 Silastic-ring method patients there were 14 cases of inadvertent ejection of a ring into the peritoneal cavity. In addition, 235 Silastic ring patients had postoperative abdominal pain severe enough to require analgesia stronger than aspirin and codeine compared to 399 of the electrocautery patients. 15 women in the electrocautery group experienced pelvic infections; there were no infections in the Silastic-ring group. Mesosalphinx bleeding occurred in both groups and most frequently with coincident suction D and C. There were cardiac complications curing the sterilizations; 6 bradycardia cases, 10 arrhythmia cases secondary to over-distention of the abdomen with carbon dioxide. Technical failure was related to previous abdominal surgery due to the presence of adhesions. 37.4% of 366 patients noted increased flow and/or duration of menses and 15% complained of menstral cramping. The majority of women reported unchanged or improved sexual relations; improved sexual relations were more significant among Silastic-ring patients.