Effect of trehalose- and sucrose-based extenders on equine sperm quality after vitrification: Preliminary results.

There has been a lack of research into equine sperm vitrification to date, but studies of other species suggest it may have significant potential. To evaluate the impact of various cryoprotectant agents (CPA) and vitrification on equine sperm quality, a controlled study was carried out. A total of 12 ejaculates were subjected to exposure to CPA and vitrification. Sperm was diluted in a range of CPA: fresh, control (BSA), sucrose (0.15M, 0.3M and 0.5M), trehalose (0.15M, 0.3M and 0.5M) and the combination of sucrose and trehalose (M1: 0.15M sucrose+0.5M trehalose; M2: 0.5M sucrose+0.15M trehalose). Sperm motility, viability, acrosome integrity and DNA fragmentation were assessed at the time of CPA exposure and after vitrification. The exposure of spermatozoa to various concentrations of sucrose and/or trehalose significantly reduced sperm motility, with lower concentrations resulting in higher sperm motility. Sperm viability and DNA fragmentation did not vary after exposure to CPA, but acrosome integrity fell significantly when spermatozoa were exposed to CPA with high osmolality. When spermatozoa were vitrified, motility values were significantly higher than those obtained during the exposure. Low concentrations of sucrose (0.15M and 0.3M) and trehalose (0.15M) showed the best progressive sperm motility. The vitrification-warmed procedure significantly reduced sperm viability and acrosome integrity, but DNA did not vary with any of CPA used. Equine sperm vitrification demonstrates a low capacity for preserving sperm motility, and extenders containing trehalose or sucrose at lower concentrations are associated with a better protective effect on sperm motility. After vitrification, acrosome and plasma membranes were severely impaired, while the DNA structure was maintained. Equine spermatozoa partially recover the motility after vitrification, but there is a need for further studies into the preservation of sperm membranes.

Short-term tolerance of equine spermatozoa to various abiotic factors.

The aim of this study was to determine the effects of various abiotic factors, such as light, physical stress (pipetting) and thermal shock, on the quality of fresh and cooled equine sperm. In experiment I, four sperm aliquots were subjected to different light exposures: (i) protected control samples (CTRL), (ii) exposed to UV light at 10 cm (UV10), (iii) exposed to UV light at 20 cm (UV20) and (iv) exposed to laboratory lighting (LAB). In experiment II, four semen aliquots were subjected to repeated pipetting for 0, 10, 20 and 30 times (CTRL, P10, P20 and P30, respectively). In experiment III, four semen aliquots at 15°C were subjected to thermal oscillations: (i) cooled control sperm at 15°C (CTRL), (ii) oscillations of 1.9°C/min to a temperature of 30°C (T30), (iii) oscillations of 1.4°C/min, with the temperature rapidly falling until reaching 1.3°C (T0R) and (iv) oscillations of 1.1°C/min, with the temperature slowly falling until reaching 4.2°C (T0S). The results revealed that after 30 min, UV10 and UV20 sperm samples showed significantly (p < .05) lower total and progressive motility values, sperm kinematic parameters and mitochondrial potential. After 45 min of exposure, differences were highly significant (p < .001). No significant differences (p > .05) were found for pipetting or thermal oscillations. The results suggest that, even if equine sperm samples are not handled in the laboratory under optimal conditions, fresh and cooled equine spermatozoa are able to resist the impact of various abiotic stimuli without any reduction in their quality. This study analyses the effect on normospermic samples, but future research could look at the tolerance that asthenozoospermic equine samples have to these abiotic influences.

Human decidual stromal cells secrete soluble pro-apoptotic factors during decidualization in a cAMP-dependent manner.

STUDY QUESTION: Is there a relationship between decidualization and apoptosis of decidual stromal cells (DSC)? SUMMARY ANSWER: Decidualization triggers the secretion of soluble factors that induce apoptosis in DSC. WHAT IS KNOWN ALREADY: The differentiation and apoptosis of DSC during decidualization of the receptive decidua are crucial processes for the controlled invasion of trophoblasts in normal pregnancy. Most DSC regress in a time-dependent manner, and their removal is important to provide space for the embryo to grow. However, the mechanism that controls DSC death is poorly understood. STUDY DESIGN, SIZE, DURATION: The apoptotic response of DSC was analyzed after exposure to different exogenous agents and during decidualization. The apoptotic potential of decidualized DSC supernatants and prolactin (PRL) was also evaluated. PARTICIPANTS/MATERIALS, SETTING, METHODS: DSC lines were established from samples of decidua from first trimester pregnancies. Apoptosis was assayed by flow cytometry. PRL production, as a marker of decidualization, was determined by enzyme-linked immunosorbent assay. MAIN RESULTS AND THE ROLE OF CHANCE: DSCs were resistant to a variety of apoptosis-inducing substances. Nevertheless, DSC underwent apoptosis during decidualization in culture, with cAMP being essential for both apoptosis and differentiation. In addition, culture supernatants from decidualized DSC induced apoptosis in undifferentiated DSC, although paradoxically these supernatants decreased the spontaneous apoptosis of decidual lymphocytes. Exogenously added PRL did not induce apoptosis in DSC and an antibody that neutralized the PRL receptor did not decrease the apoptosis induced by supernatants. LIMITATIONS, REASONS FOR CAUTIONS: Further studies are needed to examine the involvement of other soluble factors secreted by decidualized DSC in the induction of apoptosis. WIDER IMPLICATIONS OF THE FINDINGS: The present results indicate that apoptosis of DSC occurs in parallel to differentiation, in response to decidualization signals, with soluble factors secreted by decidualized DSC being responsible for triggering cell death. These studies are relevant in the understanding of how the regression of decidua, a crucial process for successful pregnancy, takes place. STUDY FUNDING/COMPETING INTERESTS: This work was supported by the Consejería de Economía, Innovación y Ciencia, Junta de Andalucía (Grant CTS-6183, Proyectos de Investigación de Excelencia 2010 to C.R.-R.) and the Instituto de Salud Carlos III, Ministerio de Economía y Competitividad, Spain (Grants PS09/00339 and PI12/01085 to E.G.O.). E.L.-D. was supported by fellowships from the Ministerio de Educación y Ciencia, Spain and the University of Granada. The authors have no conflict of interest.

Evaluation of testicular echotexture with Ecotext as a diagnostic method of testicular dysfunction in stallions.

This study aimed to assess if Ecotext, a new software for evaluation of testicular echotexture, is a good method for diagnosis of stallions with testicular dysfunction (TD). Relationships between Ecotext parameters and sperm motility and production, testicular volume, and testicular blood flow were also studied. Ecotext provides a total of six echotexture parameters: Ecotext 1 (black pixels), 2 (white pixels) and 3 (grey pixels), and another 3 parameters related to hypoechogenic areas: Ecotext tubular density (ETD), Ecotext tubular diameter (ETd), and Ecotext tubular area (ETA). Stallions (n = 33) were assessed using proven diagnostic techniques (spermiogram, B-mode and Pulse Doppler ultrasound), and subsequent analysis with Ecotext. Animals were classified as "control stallions" (n:21, acceptable semen quality), and "stallions with TD" (n:12, poor semen quality (TM < 60%, PM < 45% and total nº of sperm with PM < 2000 × 106 spz), that were subdivided into "induced TD group" (immunized, anti-GnRH vaccine) and "acquired TD group". The acquired TD group showed differences in all Ecotext parameters in relation to controls (Ecotext 1:0.11 ± 0.17 vs 2.82 ± 2.52, Ecotext 2:1584.0 ± 575.8 vs 388 ± 368.2, Ecotext 3:134.2 ± 9.26; ETA: 2.14 ± 0.59 vs 5.40 ± 1.90; ETd: 65.66 ± 6.27 vs 86.93 ± 10.65 and ETD: 92.35 ± 11.24 vs 132.10 ± 16.35, p ≤ 0.001). Results suggest acquired TD stallions were suffering testicular degeneration with loss of architecture and function as all Ecotext parameters were altered in relation to controls. Induced TD horses only showed a reduction in ETD (116.2 ± 8.59 vs 132.10 ± 16.35, p ≤ 0.001), despite all sperm parameters being worse. These findings suggested immunized stallions probably only experience an acute loss of testicular functionality and parenchyma architecture is likely not affected since differences in Ecotext parameters with control stallions were not detected. ETD was the best parameter to identify animals with TD (AUC: 0.84, optimal cut-off value of 124.3 seminiferous tubules/cm2). Correlations were found between ETD and Doppler indices (PI: 0.60; RI: 0.47 p ≤ 0.001), total testicular volume (r: 0.48; p ≤ 0.05) and sperm motility (TM:0.51; and PM:0.54; p ≤ 0.001) and production (r:0.51; p ≤ 0.001). In summary, Ecotext could identify changes in testicular echotexture of stallions with TD. Results open the possibility for new research focused on establishing the relationship between Ecotext parameters and histomorphometry features in stallion testes.

Portoenterostomy diversion. Experimental study in antireflux technique.

Ascending cholangitis was induced in dogs by performing a biliodigestive Roux-Y anastomosis. Then two valvular antireflux mechanisms were performed on separate groups of these dogs with the aim of preventing the onset of ascending cholangitis. One was performed by a laterolateral plicature at the intestinal anastomosis and the other by invagination of the mucosa in the nonworking loop that had been anastomosed to the bile duct. All the dogs underwent analytic tests over a period of 3 months and histopathologic tests at the end of the study period. Results showed cholangitis and pericholangitis in the liver biopsy specimens of the group with no antireflux valve, to a lesser degree in the group with laterolateral plicature, and almost none in the animals with the invaginated valve.

Transperitoneal absorption of intralipid in rats: total serum fatty acids and triglyceride after absorption.

Intralipid 20% was injected percutaneously into the peritoneum of 58 female rats. The rats were divided into seven groups (with an additional control group of 35 rats). To measure transperitoneal absorption, we determined the serum fatty acid and triglyceride concentrations at 3-hr intervals for 24 hr, and found a considerable increase in all of the levels measured, with a maximum at about 6 hr. The serum triglyceride levels never rose above a mean value of 200 mg/100 ml. A second and smaller rise was seen after 15 hr, declining again to the initial values. The relative proportions of the different fatty acids changed, but not drastically.

Early age thermal conditioning immediately reduces body temperature of broiler chicks in a tropical environment.

Early age thermal conditioning (TC) durably improves resistance of broilers to heat stress and reduces body temperature (Tb). Three experiments on broiler chicks were conducted to evaluate the effects of TC at 5 d of age on Tb variation measured by thermometer between 4 and 7 d of age, under a tropical environment. Because manipulation of chickens to measure Tb with a thermometer may increase Tb, a preliminary experiment on 13 3-to-4-wk-old male broilers compared Tb measured by telemetry to Tb measured in the terminal colon during three successive periods at 22, 33, and 22 degrees C. During heat exposure, Tb rapidly increased by 0.9 degrees C and plateaued over 24 h. During the last period, seven of the broilers rapidly reduced Tb to a plateau lower than the initial Tb, although six broilers exhibited more variable Tb. Measurement by thermometer underestimated on average core Tb by 0.28 degrees C at 22 degrees C and by 0.57 degrees C at 33 degrees C, whereas Tb recorded by telemetry was not affected by manipulation of the chickens. TC reduced Tb 24 h later in the three experiments. Compared to unexposed control chicks (N), 12 h of TC at 40 degrees C did not significantly reduce Tb at 7 d of age, although 24 h did. TC at 38 and 40 degrees C over 24 h significantly reduced Tb variation from 4 to 7 d of age compared to N chicks, whereas 36 degrees C did not. Withdrawing feed from the chicks for 2 h prior to measurement did not significantly reduce Tb at 4 and 7 d of age, but Tb reduction due to TC was greater in fed chicks (0.28 degrees C) than in chicks without feed (0.05 degrees C). Early age thermal conditioning at 38 to 40 degrees C at 5 d of age for 24 h reduced body temperature of 7-d-old male broilers.

[Carcinomatous lymphangitis of the lung. Report of 21 cases (author's transl)]. / Linfangitis carcinomatosa pulmonar. Estudio de 21 casos.

Twenty-one cases of carcinomatous lymphangitis of the lung are reported. In diagnosing them clinical, roentgenologic, and histopathologic criteria were used and the clinical, roentgenologic and evolutive findins were considered in order to emphasize the origin of the primitive neoplasm. From this point of view the principal ones turned out to be carcinomas of the breast, stomach, and lung. The absence of carcinoma of the thyroid gland as a source of pulmonary lymphangitis carcinomatosa must be pointed out. The higher incidence of this condition in older people and the clear predominance in the female sex are stressed. From a clinical standpoint the most important data were the existence of cough and dyspnea, which are often associated with the commom presence of enlarged left supraclavicular or axillary lymph nodes. There was a predominant pure interstitial pattern (the most common feature was the presence of Kerley's A or B lines), sometimes coexisting with alveolar impairment and very often with accompanying pleural effusion. The mean survival rate was 2 months, with a maximum of 7 months and a minimum of 3 days. The results of the present series were compared with those of the literature and the most significant and constant fact in both instances was that two thirds of the cases of pulmonary lymphangitis carcinomatosa had their origin in neoplasms of the breast, stomach, and lung.

Use of serial ultrasound measures in the study of growth- and breed-related changes of ultrasonic measurements and relationship with carcass measurements in lean cattle breeds.

The growth and breed-related changes of rib and rump characteristics in lean beef cattle and the ability of ultrasound to predict carcass traits were investigated. Three hundred bulls from three breeds were scanned monthly (6-7 times) using real-time ultrasound with final scans taken <7 days prior to harvest. The rib and rump ultrasound measurements, except intramuscular fat content, increased (P<0.05) with live weight. Breed affected most of traits. The 12th rib ultrasound measurements showed a weak to high positive correlation (0.131 to 0.976, P>0.05 to P<0.001) with 12th rib measurements in the carcass. Regression equations developed with the ultrasound measurements, explained 97% of the variation in longissimus muscle area, 88% of the variation in fat thickness and 57% of the variation in intramuscular fat content. When last ultrasound scan measurements were excluded from prediction equations, the R(2) significantly decreased. Ultrasound measures "in vivo" are viable options for assessing carcass attributes of lean cattle.

Functional anatomy of the medial collateral ligament of the ankle joint.

The calcaneotibial and posterior talotibial ligaments slacken and the naviculotibial ligament tightens as the ankle plantarflexes; the reverse occurs in ankle extension. The naviculotibial ligament increases its length and the posterior talotibial ligament relaxes in abduction. The cutting of the posterior talotibial ligament repercutes on other parts of the medial collateral ligament. The cutting of other parts of the medial collateral ligament produces very little change on the posterior talotibial ligament. When the whole medial collateral ligament is severed, there is a lateral displacement of the talus. This may be important in the diagnosis and surgical treatment of a torn medial collateral ligament.

Effect of exogenous serotonin on intragastric pH and its influence on serum gastrin levels in rats.

The effect of various doses of serotonin on the serum gastrin levels and intragastric pH in rats, was studied. After serotonin administration of 10 mg/kg i.p., a significant increase in serum gastrin levels was noted, as well as a strong increase in the intragastric pH. It was also observed that a lower dose, 5 mg/kg i.p., significantly increased serum gastrin levels, while intragastric pH was not affected, remaining at baseline values throughout the study. These results suggest that the increase observed in serum gastrin levels after administration of exogenous serotonin is not mediated by increase in intragastric pH.

Possible mechanism of stimulation of gastrin secretion by exogenous serotonin in rats.

The effect of 6-hydroxydopamine, propranolol, phentolamine, alpha-methyl-tyrosine and alpha-methyl-tyrosine plus propranolol on serotonin-stimulated gastrin secretion in rats has been examined. Gastrin secretion in response to administration of serotonin alone (10 mg/kg i.p.) was significantly reduced in rats pretreated with 6-hydroxydopamine or with propranolol. These results suggest that the effect of exogenous serotonin on gastrin secretion can be described as sympathomimetic and indirect. The serotonin-stimulated gastrin secretion was significantly enhanced by previous administration of phentolamine. Pretreatment with alpha-methyl-tyrosine also elevated serotonin-stimulated gastrin secretion, indicating that in the presence of diminished concentrations of the catecholamines, the influence of exogenous serotonin on secretion by G cells is increased. This enhancement in the serum gastrin levels was also reduced to a significant extent by simultaneous administration of propranolol, which suggested the activation of G-cell beta-adrenergic receptors after serotonin administration.

[Biochemical complications of experimental vesico-sigmoidostomy]. / Complicaciones bioquímicas de la vesicosigmoidostomía experimental.

Three experimental models of vesico-sigmoidostomy are studied (model-1, end to side V-S plus urethral ligation, model-2, end to end V-S, in "Y of Rous" plus urethral ligation and model-3, vesico-sigmoidoplasty), with aim of reproducing chemical imbalance observed in human subjects with ureterosigmoidostomy. Authors have evaluated clinical biochemical (serum acido-base balance, Cl, Na+, K+, BUN, creatinine, ammonia and albumin), and histologic variables in the first, third and fifth month after operation in 225 rats. Animal of model-1 presented more frequently than model-2 and model-3, alterations (hyperchloraemic acidosis, uraemia, hyperammonemia and hypoalbumin) as well as affectation of upper urinary system for acute or chronic pyelonephritis.

Hemangioendotelioma epitelioide pulmonar: causa inhabitual de supuestas metástasis pulmonares múltiples de primario desconocido / No disponible

El hemangioendotelioma epitelioide pulmonar es un raro tumor de estirpe vascular endotelial, de bajo-intermedio grado de malignidad y con comportamiento clínico impredecible. Afecta principalmente a mujeres en edad media de la vida, con escasos síntomas en el momento del diagnóstico que suele ser incidental al realizar una exploración radiológica por otro motivo. Suele cursar con nódulos pulmonares múltiples bilaterales, sin afectación ganglionar mediastínica ni pleural, que obliga al diagnóstico diferencial principalmente con enfermedad metastásica e inflamatoria granulomatosa. Aunque la tomografía por emisión de positrones (PET) puede ayudar a discernir el carácter tumoral de las lesiones, es necesario el estudio histológico tras toracotomía, en el que las técnicas de inmunohistoquímica definen el carácter vascular. El pronóstico es incierto y el tratamiento no está estandarizado. Se contempla la exéresis quirúrgica en las formas limitadas principalmente, con escasos resultados en las formas diseminadas al tratamiento quimioterápico y experiencias escasas con antiangiogénicos (AU)

Pulmonary epithelioid hemangioendothelioma is a rare vascular tumor of endothelial lineage, low-intermediate grade malignancy and unpredictable clinical behavior. It mainly affects middle-aged women of life, with few symptoms at the time of diagnosis is usually incidental when making a radiological for another reason. Usually present with multiple bilateral pulmonary nodules without mediastinal lymph node or pleural, forcing the differential diagnosis with metastatic disease primarily granulomatous inflammatory. Although positron emission tomography (PET) may help to discern the nature of the injuries tumor, histologic examination is necessary after thoracotomy, in which immunohistochemistry vascular define the character. The prognosis is uncertain and treatment is not standardized. Surgical excision is contemplated in the limited ways mainly to poor results in disseminated forms to chemotherapy and antiangiogenic drug scarce experiences (AU)