Overexpression of Hypoxia-Inducible Factor-1α in Primary Graft Dysfunction Developing in an Orthotopic Lung Transplantation Rat Model.

BACKGROUND: Primary graft dysfunction (PGD) is the major cause of early morbidity and mortality after transplantation. A high rate of PGD is a frequent complication in orthotopic lung transplantation (OLT) models, which are currently used to investigate acute and chronic rejection pathways. Hypoxia-inducible factor (HIF)-1α is a heterodimeric αß transcription factor that mediates tissue response to hypoxia. In other solid organ transplantations, a significant correlation between HIF-1α expression and PGD was detected. To our knowledge no data are available on HIF-1α expression in PGD developing in lung transplantation. The aims of this study were to investigate HIF-1α expression (using immunohistochemistry) and correlate it to the main histological parameters related to ischemia-reperfusion (IR) injury, including terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) -positive apoptotic cells). METHODS: OLT was performed in 32 inbred rat strains and 11 of them died in the early postoperative period (from day 0-3) for IR injury. The histological and molecular evaluations were done in all lung tissues. Unimplanted donor rat lungs were used as controls. HIF-1α expression was correlated with all morphological parameters. RESULTS: Lung samples of animals with IR injury showed high scores of HIF-1α expression, edema, blood extravasation, granulocyte margination, apoptotic index, and necrosis in 91% of cases. Tissue overexpression of HIF-1α was detected in all lung samples with high scores of histological parameters and with high apoptotic indexes. CONCLUSION: Our data demonstrate that HIF-1α was overexpressed in more severe rat lung IR injury. The use of HIF-1α inhibitors could provide a translatable route into manipulating this complex system in vivo.

Pain alleviation during castration of piglets: a comparative study of different farm options.

Pain alleviation associated with castration of piglets is an important welfare issue. The present study compares the effect of different approaches and products suitable for farmer use, with the aim to alleviate pain due to castration in piglets. A randomized within-litter design, with 28 replicate litters, compared 7 treatments: handling () restraint of the piglet and manipulation of the scrotum, castration without pain relief (), 2 treatments (, ) with different concentrations of tetracaine (2 and 6%) applied topically 10 min before and immediately post-surgery, and 3 treatments with i.m. injection of different nonsteroidal anti-inflammatory drugs () 10 min prior to surgery (-meloxicam, -ketoprofen, -tolfenamic acid). Efficacy of pain relief was assessed during a 300 min period after castration by serum cortisol, behavior (walking, lying, suckling, in the nest, isolated and pain related: tremors, rubbing the rear, hunching, wagging of the tail), facial expression and scrotal skin pressure sensitivity. C pigs had greater serum cortisol concentration than all other groups at 60 min post-surgery ( < 0.001), while H pigs had lower concentrations than pigs given topical anesthesia ( < 0.001) though not injected analgesia. No treatment differences were significant at 180 min, but at 300 min cortisol concentration was greater in T2 and T6 piglets than those given NSAIDs ( = 0.03). These treatment differences were mirrored by the pressure sensitivity of the scrotum; in comparison with C piglets, those given NSAIDs showed a reduced sensitivity ( 0.003) but those given local anesthesia did not ( = 0.15). C pigs showed increased frequency of pain-related behavior in the first 30 min in comparison with all other treatments, more time isolated than H or NSAID treatments, and more time standing inactive than H or K treatments. No behavioral differences were apparent after 60 min. No differences in facial expressions were observed among treatments. In conclusion, on-farm methods for pain relief can provide some, though not complete, pain alleviation in the hours after castration. The use of topical anesthesia gave only minor benefit in comparison to NSAID agents injected prior to castration. Since the main differences in indicators of pain between positive and negative controls were observed within the first h after castration, it is important to select drugs that act quickly after administration to facilitate practical processing schedules on farm.

Two-dimensional, M-mode and Doppler-derived echocardiographic parameters in sedated healthy growing female sheep.

Despite the fact that sheep are a widely used animal model in cardiovascular research, reference values for transthoracic echocardiography in normal growing animals are not available. Eight healthy female lambs underwent two-dimensional, M-mode and pulsed wave Doppler echocardiographic examination at 100 days of age and every three months thereafter over a 12-month period. The study was conducted under sedation with midazolam, butorphanol and constant rate infusion of intravenous propofol. Their growth phase was completed at about one year of age. All the echocardiographic parameters considered were significantly correlated with body weight and age class except for the left ventricular systolic and diastolic diameters. Functional indices were not correlated to body weight or age except for the E-point to septal separation distance (EPSS). Doppler-derived parameters were not influenced by independent variables. Transthoracic echocardiography can be considered an applicable method for cardiovascular research using a growing lamb animal model after appropriate adjustments for age and body size.

The effects of different environmental conditions on thermoregulation and clinical and hematological variables in long-distance road-transported calves.

The aim of this study was to investigate the effects of long-distance road transport (19 h, from Poland to Italy) during 2 seasons (summer vs. winter) on clinical and hematological variables in calves. The environmental temperature range that could compromise the thermoregulation system (thermal stress) of the calves was tested. For the 7 Holstein calves in each transport, the BW and rectal temperature (RT) were measured, and blood samples were collected at the farm of origin, before loading at the transit center (T2), after unloading at the farm of destination (T3), and 1, 2, 3, and 4 d after arrival. The body temperature (BT) and heart rate (HR) were continuously monitored from T2 to T3. The data were statistically analyzed according to a mixed model that considered the fixed effects of transport (repeated measurements), season of journey, and their interaction. Within the observed temperature-humidity index (THI) range (30 to 80), effective thermoregulation allowed the calves to maintain their BT with small physiologic changes to prevent thermal stress, particularly in the summer. With no seasonal differences, the HR was greater at loading than unloading (120 vs. 115 beats per min; P = 0.012). As for the transport effect, the BW was less (P < 0.001) after unloading, and the RT was greater (P = 0.004). This effect was more marked in summer. The hematological variables indicated a moderate effect of transport on the hydration condition, reactive and muscular systems, and metabolism, although hematocrit (P = 0.004), erythrocytes, cortisol, NEFA, ß-hydroxybutyrate, lactate, creatine kinase, lactate dehydrogenase, and aspartate aminotransferase activity (P < 0.001), and total protein (P = 0.007) were greater after unloading. This was confirmed by a moderate decrease in total leukocytes (P = 0.031) and glucose concentration (P = 0.002). The changes in the clinical variables were similar for both seasons even though in the summer, hematocrit (P < 0.001), urea (P = 0.008), and total protein (P = 0.010) increased and glucose concentration (P = 0.038) decreased. In conclusion, the data did not show a pronounced effect attributable to the season of the journey. Long-distance road transport leads to notable changes in clinical and hematological variables at the end of the journey. However, these variables remained within their physiological ranges and returned to basal values within a few days after the journey.

Nerve growth factor concentrations in the synovial fluid from healthy dogs and dogs with secondary osteoarthritis.

OBJECTIVE: To measure the concentrations of nerve growth factor (NGF) in the synovial fluid from normal dogs and dogs with osteoarthritis (OA) secondary to common joint disorders. METHODS: Nerve growth factor synovial concentrations were measured by ELISA assay in 50 dogs divided into three groups: 12 healthy, 16 affected by acute lameness within seven days before enrolment, and 22 with chronic lameness persisting by more than one month before enrolment and accompanied by radiological signs of OA. Both acute and chronic lameness were secondary to orthopaedic diseases involving the shoulder, elbow and stifle joints. Nerve growth factor synovial concentrations were compared between means for healthy and acute groups and between the three groups using an F-test. Significance level was set at p <0.05. RESULTS: Nerve growth factor was detected in all canine synovial fluid samples. However, the mean synovial NGF concentration of healthy dogs (3.65 ± 2.18 pg/ml) was not significantly different from the mean value in dogs with acute lameness (6.45 ± 2.45 pg/ml) (p = 0.79). Conversely, the mean synovial NGF concentration in dogs with chronic lameness (20.19 ± 17.51 pg/ml) was found to be significantly higher than that found in healthy dogs (p <0.01). CLINICAL SIGNIFICANCE: This study demonstrates for the first time the presence of NGF in canine synovial fluid and its increased concentrations in dogs with chronic lameness compared to healthy dogs and dogs with acute lameness. The association between chronic lameness and raised synovial concentrations may suggest an involvement of NGF in OA inflammation and chronic pain.

Relationship between milk fatty acid composition and dietary roughage source in dairy cows.

According to a 2 x 2 crossover design, 14 Holstein dairy cows were fed two isoenergetic diets based on either grass hay (GH) or maize silage (MS). Milk samples were collected during the third week of each period, and fatty acid (FA) profiles were analyzed using gas chromatography. The data obtained were subjected to ANOVA. Dietary treatment had no effect on either dry matter intake or milk yield. Milk from animals fed the GH-diet contained lower concentrations of saturated FAs (61.9 vs. 63.4% of total FAs; P < 0.05) and higher levels of polyunsaturated FAs (PUFAs) (6.1 vs. 5.8; P < 0.01). Feeding additional hay also increased conjugated linoleic acid and n-3 FA levels and decreased C16:0 levels. Increases in both PUFAs and n-3 FAs resulted in lower (P < 0.01) atherogenic and thrombogenic indices in milk from animals fed the GH diet compared with those fed the MS diet. A complete substitution of GH for MS appeared to improve milk FA profiles, even using different types of concentrates to provide a balanced diet.

Serum biochemistry and electrophoretic patterns in the Eurasian Buzzard (Buteo buteo): reference values.

In avian medicine, hematologic and biochemical laboratory investigations are still in their infancy, because of the difficulty involved in collecting data. This has led to a lack of reference values and a nonstandardized approach to specimens obtained in critical conditions. The Eurasian Buzzard (Buteo buteo) is one of the most common raptors in Italy, yet little is known about the physiologic blood parameters of this species. Serum biochemistry and electrophoretic investigations were performed in 40 healthy Eurasian Buzzards in different Italian wildlife rescue centers waiting to be released after recovering from trauma injuries. Mean values for biochemistry parameters were pancreatic amylase 626.9 IU/l, uric acid 7.5 mg/dl, aspartate aminotransferase 330.9 IU/l, glucose 375.1 mg/dl, lipase 26.3 IU/ l, total protein 38.4 g/l, total bilirubin 0.04 mg/ dl, lactate dehydrogenase 2,008.4 IU/l, creatinine kinase 1,604.1 IU/l, alanine aminotransferase 40.4 IU/l, alkaline phosphatase 89.8 IU/l, magnesium 2.3 mg/dl, calcium 10.2 mg/dl, phosphorus 2.02 mg/dl, cholesterol 192.2 mg/ dl, triglyceride 116.4 mg/dl, albumin 14.5 g/l, creatinine 0.1 mg/dl. Mean electrophoretic values were prealbumin 1.4 g/l, albumin 14.2 g/l, alpha 1 globulin 5.9 g/l, alpha 2 globulin 4.7 g/l, beta globulin 7.5 g/l, gamma globulin 3.6 g/l, albumin/globulins ratio 0.8 g/l.

¿Son modificables las actitudes hacia laspersonas mayores en estudiantes de medicina?Experiencia en la Universidad de Salamanca / Are the medical students’ attitudes towards older people modifiable? The University of Salamanca’s experience

Introducción. Los médicos y los estudiantes de medicina tienden a tener actitudes negativas hacia las personas mayores. En la Unión Europea, sólo el 50% de los estudiantes de medicina reciben formación en medicina geriátrica. Sujetos y métodos. Se evaluaron las actitudes hacia las personas mayores en la cohorte de estudiantes de medicina de 5.º del curso 2006-07 inscritos en la asignatura de geriatría, al comienzo y al fin de la misma. Se utilizó un cuestionario basado en la escala diferencial semántica de Rosencranz et al, donde las puntuaciones bajas indican actitudes más positivas. Resultados. Se obtuvieron datos de 93 estudiantes, 74% fueron mujeres y 26% varones. El promedio de edad fue 22,4 años. Se apreció una mejoría en la mayoría de las variables evaluadas, con diferencias significativas en 21 de las 32 variables analizadas. Sólo el15% declaró haber recibido un entrenamiento previo en el área del envejecimiento. Conclusiones. La exposición a conocimientos y prácticas basadas en la atención y cuidado de las personas mayores modificarían positivamente las actitudes de los estudiantes de medicina (AU)

Introduction. Both physicians and medical students show negative attitudes towards older people. In the European Union, only about 50% of the medical students receive geriatric medicine training. Subjects and methods. Attitudes towards elder people were evaluated in medical students in the fifth year of their studies during 2006-07 academical year, before and after receiving specific training in geriatric medicine. To measure attitudes we used the instrument developed by Rosencranz et al, which incorporates the Aged Semantic Differential; low scores show positives attitudes. Results. 93questionnaires were filled in correctly; 74% of the participants were women and 26% men. Average age was 22.4 years. Our results shows lower values in the majority of the 32 variables studied, with statistically significant differences in 21 of them. Only 15% of the participants declared to have received geriatric medicine training before our study. Conclusions. Medical students’ exposure to knowledge and practice in elderly care could positively modify their attitudes (AU)

Identificación de mayores con trastornos mentales y su relación con autopercepción de salud en medio comunitario rural. Estudio de corte transversal / Identification of elderly subjects with mental illness and its relationship with self-perception of health in a rural community setting. A cross-sectional study

INTRODUCCIÓN. El incremento de la carga de los trastornos mentales en la población de mayores y los pocos datos de prevalencia en medio rural comunitario son la razón de esta comunicación. Nuestro objetivo fue identificar adultos mayores con trastornos neuropsiquiátricos (ansiedad, depresión, distimia) en medio comunitario rural y su relación con la autopercepción del estado de salud. MATERIAL Y MÉTODOS. Estudiamos una cohorte de mayores de 65 años (n = 165) de Garcibuey (C1) y Villanueva del Conde (C2), dos pueblos de la provincia de Salamanca. Utilizamos un cuestionario realizado en base al de la Organización Panamericana de la Salud/Organización Mundial de la Salud (OPS/OMS) para el estudio SABE que revela aspectos en 4 dominios diferentes: datos demográficos, estado de salud, estado nutricional y aspectos sociales de los adultos mayores. Los datos se obtuvieron entre noviembre de 2006 y febrero de 2007 por entrevista directa y revisión sistemática de historias clínicas. Utilizamos para el tratamiento estadístico el paquete SPSS 12.0. RESULTADOS Y CONCLUSIONES. Cuarenta y una (25,2%) de las personas han sufrido o sufren un trastorno neuropsiquiátrico, 121 (74,2%) no. Nueve están bajo tratamiento médico farmacológico. El 63,4% vive solo o con su pareja; 22 (53,6%) perciben su estado de salud como malo o regular. El 65,8% consume psicofármacos. Las mujeres (28 [68,2%]) fueron las más afectadas (AU)

INTRODUCTION. This paper aims to deal with the increaseof the burden of mental disorders in the elderly populationand scarce prevalence data in the rural communitysetting and the poor evidence in rural settings. Our aim wasto identify elderly adults with neuropsychiatric morbidity(anxiety, depression and dysthymia) in rural communityarea, and their relationship to self-perception of one’s healthcondition.MATERIAL AND METHODS. A cohort made up of subjectsover 65 years (n = 165) from Garcibuey (C1) and Villanuevadel Conde (C2), Salamanca, was studied. We used aquestionnaire based on the SABE study from Pan-AmericanHealth Organization/World Health Organization (PAHO/WHO) to evaluate personal data, health and nutritionalstatus and social issues in elderly subjects. Data was collectedbetween November 06 - February 07 by direct personalinterview and systematic review of medical record.RESULTS AND CONCLUSIONS. Forty-one (25.2%) ofthe cohort have suffered or are suffering a neuropsychiatricdisorder, 9 of whom are currently under medical treatment.A total of 63.4% live alone or with a partner; 22 (53.6%) perceivetheir health status as fair or bad. A total of 65.8% consumepsychodrugs. Neuropsychiatric disorders are more frequentin women (28 [68.2%]) (AU)

A saturated N-acylethanolamine other than N-palmitoyl ethanolamine with anti-inflammatory properties: a neglected story...

N-acylethanolamines, which include the endocannabinoid anandamide and the cannabinoid receptor-inactive saturated compounds N-palmitoyl ethanolamine and N-stearoyl ethanolamine, are ethanolamines of long-chain fatty acids degraded by fatty acid amide hydrolase (FAAH) known to accumulate in degenerating tissues and cells. Whilst much evidence supports a protective anti-inflammatory role of both anandamide and N-palmitoyl ethanolamine, very little information is available with regard to the bioactivity of N-stearoyl ethanolamine. Employing a murine model of passive IgE-induced cutaneous anaphylaxis, we have found that N-stearoyl ethanolamine is endowed with marked anti-inflammatory properties in vivo, supporting the hypothesis that endogenous N-stearoyl ethanolamine is, in analogy to N-palmitoyl ethanolamine, a bioactive signalling lipid capable of downregulating allergic inflammation in the skin. This effect, although mimicked by synthetic, non-selective, CB(1)/CB(2) receptor agonists, such as WIN55, 212-2, was not sensitive to CB(1) or CB(2) receptor antagonists, but rather was fully reversed by capsazepine, a competitive antagonist of the TRPV1 receptor. Moreover, CB(1) receptor antagonists, although effective in antagonising the WIN55,212-2-induced hypothermia, did not reduce the anti-inflammatory effect of WIN55,212-2, whilst CB(2) receptor antagonists, per se inactive, potentiated the WIN55,212-2 effect, suggesting an involvement of non-CB(1)/CB(2) receptors in the anti-inflammatory action of WIN55,212-2. All this, together with demonstration of FAAH as a major regulator of the in vivo concentrations of saturated N-stearoyl ethanolamine, in addition to N-palmitoyl ethanolamine, raise the speculation that pharmacological treatments with saturated N-acylethanolamines such as N-stearoyl ethanolamine, or alternatively FAAH inhibitors able to increase their local concentration, rather than selective CB receptor agonists, might be of promising therapeutic benefit in reducing allergic inflammation in the skin.

Validation of an automated spectrophotometric assay for the determination of cholinesterase activity in canine serum.

The determination of enzymatic activity of cholinesterase is a useful diagnostic method to detect exposure to anticholinesterase compounds in human and in veterinary medicine. We validated a modification of the Ellman method in canine serum and applied it to the diagnosis of dogs poisoned with anticholinesterase substances. The method used butyrylthiocholine as substrate and potassium hexacyanoferrate as chromophore. The reference range calculated on 60 clinically healthy dogs was set between 3405 and 6561 U/L (chi-square test for normal distribution, p > 0.05). The overall mean intra-assay and inter-assay coefficients of variation were 0.53% and 3.83%, respectively. The assay was linear when using two sera with 12,538 U/L and 6604 U/L serum cholinesterase activity (r(2) = 0.997) and 0.999, respectively). The mean recovery values of pooled sera with a mean pseudocholinesterase (PChE) activity of 12,081 U/L and pooled sera with a mean PChE activity of 3415 U/L were 103.5% and 102.8%, respectively. Six dogs with a diagnosis of anticholinesterase compound intoxication showed a decrease in cholinesterase activity of at least 50% of normal activity with a mean +/- SD of 487 +/- 291 U/L ranging from 169 to 847 U/L. This technique conforms to the current standard for precision, linearity and accuracy and is a useful method for the complementary diagnosis of organophosphate or carbamate insecticide intoxication in dogs.

Vertebral mineral density measured by dual-energy X-ray absorptiometry (DEXA) in a group of healthy Italian boxer dogs.

We studied the feasibility of using dual-energy X-ray absorptiometry (DEXA) to obtain reference bone density values in relation to age, gender and body weight in growing and young adult Italian boxer dogs. The study was performed on eight animals (three males and five females) at 7, 12 and 18 months of age. Animals were carefully examined and blood samples were collected from each dog to detect any sign of metabolic and/or endocrine disease. Each subject underwent radiographs to evaluate growth of the spine and hip. One female was not considered in the statistical model because of the development of grade 4 spondylosis deformans during the study period. All animals were serially scanned using DEXA; the region of interest was the whole spine T12-L2, while the subregions of interest were the four vertebrae (T12-T13-L1-L2) within the scanned spine. Statistical analysis was performed separately for each region of interest. Age had the strongest relationship with bone density (P < 0.001). Gender effect on spinal mineral density was not significant while vertebral site effect was highly significant. Average bone mineral density (BMD) +/- SD for the whole spine trait was 0.862 +/- 0.108 g/cm2 while average BMD +/- SD for subregions of interest ranged from 0.836 +/- 0.141 g/cm2 for T12 to 0.928 +/- 0.119 g/cm2 for L2. Estimated reference BMD values at 7, 12 and 18 months of age for each vertebral site in males and females are provided.

Dose-finding study of weekly 24-h continuous infusion of 5-fluorouracil associated with alternating oxaliplatin or irinotecan in advanced colorectal cancer patients.

PURPOSE: To determine maximum tolerated dose, safety and efficacy of weekly 24 h infusional 5-fluorouracil (5-FU) combined alternately with oxaliplatin and irinotecan. PATIENTS AND METHODS: Advanced colorectal carcinoma patients in first- or second-line chemotherapy received increasing doses of 5-FU (weekly 24 h continuous intravenous infusion without leucovorin) on days 1, 8, 15 and 22, irinotecan days 1 and 15; and oxaliplatin days 8 and 22, every 35 days. RESULTS: Thirty-four patients received 175 cycles. The median age was 64 years (range 47-78). Eighteen per cent of patients had the primary tumor in the rectum, with a median of one disease site (range one to three), and liver involvement in 88% and lung in 38%. Six (18%) patients had chemotherapy for prior advanced disease. The most frequent grade 3-4 toxicity was neutropenia (41% of patients), but the regimen was well tolerated clinically, with febrile neutropenia in two patients and grade 4 neutropenia lasting >7 days in one; grade 3-4 diarrhea, nausea and vomiting in 6% of patients; grade 3-4 peripheral neuropathy in 9% of patients. Seventeen patients had a partial response (50%; 95% confidence interval 33%-67%), 13 had stable disease and one had progressive disease. Five patients underwent metastatic surgical resection after tumor shrinkage. Median response duration was 14 months (range 4.7-29.2+) and median time to progression was 11.3 months (range 1.1+-30.7+). CONCLUSIONS: This combination three-drug regimen is feasible and well tolerated without toxicity overlap. Preliminary antitumor activity compares well with standard double combinations, with an unusually long median time to progression. The recommended dose is 5-FU 3000 mg/m(2), weekly for 4 weeks, irinotecan 100 mg/m(2) days 1 and 15, oxaliplatin 80 mg/m(2) days 8 and 22. Further assessment of antitumor activity and safety is warranted.

Neurogenic inflammation in primary headaches.

The headache in migraine is thought to result from neuronal nociceptive activity in the trigeminovascular system, that is, the meninges. In addition, trigeminal axons projecting to the meninges contain vasoactive neuropeptides, such as substance P, calcitonin gene-related peptide and neurokinin A, that may promote, when released, plasma protein leakage and vasodilation within dura mater, characteristic of neurogenic inflammation. Thus, it has been hypothesized that a sterile neurogenic inflammation in the meninges may be involved in generating or sustaining, via occurrence of a vicious cycle, the pain accompanying the migraine attacks. We here review the evidence in support of this hypothesis as well as its potential significance in better tailoring therapies in migraine or other types of primary headaches.

[Heterotopic gastric mucosa in the upper esophagus causing dysphagia. A case report]. / L'eteroplasia gastrica come causa di stenosi dell'esofago cervicale. A proposito di un caso.

The Authors report a 10-years-old boy who was diagnosed as having upper esophageal stenosis. Biopsies of the stricture showed heterotopic gastric mucosa; endoscopic treatment was safe and effective.

A radiographic technique to assess the longitudinal balance in front hooves.

The aim of this study was to validate a radiographic technique to measure objectively the longitudinal hoof balance in the horse. Ten left third phalanges (P3) obtained from front hooves and 19 left front hooves were used in order to assess the reliability of the radiographic technique and to identify any effect of rotation or alignment of specimens or distance between the X-ray machine and the radiographic cassette, in measuring P3s structures and balance related parameters. A rotational support was made to hold specimens in order to standardise some of the conditions of X-raying. The main axis of the frog was used as a marker for the sagittal plane of P3. Results showed that single radiographic measurements could have wide limits of agreement with actual measurements of the hoof (gold standard values). In order to limit errors we suggest the use of a combination instead of single parameters to assess balance. Rotation of the hoof is also to be controlled when a measurement of angles is needed. The radiographic technique used in this study can be used to objecticely measure balance parameters considered important in order to assess hoof balance.