Transforming growth factor type beta (TGF-beta) and adipogenesis in pigs

The present study was performed on s.c. adipose tissue of fetal pigs at 35 to 110 d of gestation to examine the distribution of TGF-beta-positive cells, to localize TGF-beta immunoreactivity at the cellular level using electron microscopy (EM), and to determine the effect of TGF-beta on primary cultures of pig adipose tissue cells. Tissues for EM were fixed and embedded in LR white resin. Sections then were incubated with a polyclonal antibody specific for TGF-beta and TGF-beta was located using 20 nm colloidal gold conjugated second antibody. Tissues were fixed and embedded in paraffin for localization of TGF-beta at the light microscope (LM) level. Tissues were incubated with anti-TGF-beta followed by localization using biotinylated second antibody. Using LM, only a few cells stained positively for TGF-beta within developing blood vessels at 35 d. By 50 d, more TGF-beta-positive cells were associated with forming capillary networks. Between 70 d and 110 d, positively stained adipocytes usually were clustered around blood vessels. Cells surrounding hair follicles stained positive for TGF-beta between 90 to 110 d. Electron microscopy revealed TGF-beta labeling within fat cells. Fibroblasts and endothelial cells did not exhibit TGF-beta immunoreactivity. The addition of TGF-beta to primary cultures of s.c. adipose tissue cells from newborn pigs prevented lipid filling in fat cells. This effect was dose-dependent, with half-maximal inhibition occurring at 3 pM maximum inhibition occurred at 40 pM. These results indicate that TGF-beta may regulate angiogenic activity and lipid filling in s.c. adipose tissue of fetal pigs. Although TGF-beta was present in adipocytes and in cells associated with developing capillary networks, the physiological role of TGF-beta during early adipose tissue development is not known.     

Adaptation of 5'-deiodination to hypothyroid conditions following surgical and/or radiothyroidectomy in chickens

The effect of thyroidectomy on serum levels of thyroid hormones (thyroxine, T4; triiodothyronine, T3) and on tissue levels of T3 as well as on 5'-deiodination (5'-DI) in liver, kidney and brain of chickens was investigated. The most effective way of thyroidectomy was a combined surgical (Tx) plus radiothyroidectomy (RTx) with low amounts of 131-I (NaI). The latter destroyed extra-thyroidal hormone production. It was found that even if after Tx and RTx the serum levels of T4 and T3 were close to the detection limit, the tissue T3 level was only half of normal in liver and kidney and remained normal in the brain. It is suggested that lowered 5'-deiodination is an important but not exclusive factor contributing to this adaptation of cells to hypothyroid conditions.     

Correlation of DNA ploidy to tumor histologic grade, clinical variables, and survival in dogs with mast cell tumors.

By using flow cytometry, a retrospective analysis of the DNA content of 40 primary canine mast cell tumors and seven lymph nodes that contained metastatic mast cell tumor from 44 dogs of various breed, sex, and age was performed on formalin-fixed, paraffin-embedded samples of the tumors and nodes. These samples were chosen according to the following criteria: samples contained sufficient well-preserved tumor tissue in the paraffin block for processing, sufficient patient history data were available, clean and homogeneous cell suspensions were obtained after processing, and interpretable DNA histograms were produced on analysis. The ploidy data obtained were compared with the histopathologic grade, the anatomical site of occurrence, the clinical stage of the tumors, and the survival of the dogs. Over 70% (29/40) of the mast cell tumors were diploid. Three metastatic mast cell tumors in lymph nodes had the same ploidy status as their corresponding primary tumors. In five dogs, mast cell tumors from multiple sites in each dog displayed similar ploidy status. Of 26 dogs evaluated for survival times, 69% (18/26) had diploid tumors and 31% (8/26) had aneuploid tumors. When numbers of diploid versus aneuploid tumors were compared, no significant difference was found between any two grades, clinical stages, or anatomic sites. A significant difference (P = 0.02) was found, however, between aneuploid and diploid tumors when comparing Stage I and non-Stage I disease. The Kaplan-Meier survival plot indicated a tendency towards an increased survival within the first year in dogs with diploid versus aneuploid tumors (P = 0.06).     

Evaluation of sodium thiosulfate as an extracellular water marker in cattle.

Two studies were conducted to determine whether sodium thiosulfate (THS) can estimate extracellular water (ECW) in beef cattle in conjunction with empty body water (EBW) estimation by urea space. Experiment 1 used 24 steers (366 kg) to determine the clearance parameters for THS and urea. Blood samples were taken over 1 h. A two-component curve, Y = A1ek1(t) + A2ek2(t), (t = hours after infusion) fit the clearance of both markers; intercepts (A1, A2) and clearance coefficients (k1, k2) were 44.8, 44.4, -25.8, and -2.24 mg/dL, respectively, for THS (r2 = .98, Sy.x = 2.72, animal effects removed and 24.4, 10.5, -21.7, and -.71 mg/dL, respectively, for urea (r2 = .98, Sy.x = 1.49). Sodium thiosulfate equilibrated with ECW 5 to 10 min after infusion. Experiment 2 consisted of 22 steers (483 kg) infused with a combination solution of 20% urea, 10% THS, and 4% sodium thiocyanate (SCN; equilibration time = 28 min); half the steers were implanted with estradiol. Empty body water increased with implantation (P less than .01). Extracellular water tended to increase in implanted steers as measured by THS (12 min, P = .14) and SCN (P = .10). The estimation of ECW at 12 min was not different (P greater than .2) from the SCN estimate at 28 min (SCN = 3.7 + .873 THS; r2 = .70; P less than .001). Sodium thiosulfate gave reasonable estimates of ECW (22 to 26% of BW) and required only 0- and 12-min blood samples.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serum amyloid A protein concentration measured by radial immunodiffusion in Abyssinian and non-Abyssinian cats

Serum amyloid A (SAA) protein concentration was determined by use of radial immunodiffusion (RID) in 4 groups of cats: Abyssinian cats with amyloidosis, healthy Abyssinian cats without clinical evidence of amyloidosis, hospitalized non-Abyssinian cats, and clinically normal non-Abyssinian cats. Mean SAA concentration in Abyssinian cats with amyloidosis was significantly (P = 0.05) higher than mean SAA concentration in healthy Abyssinian cats without clinical evidence of amyloidosis and in hospitalized non-Abyssinian cats. Mean SAA concentration in clinically normal non-Abyssinian cats was significantly (P = 0.05) lower than mean SAA concentration in healthy Abyssinian cats without clinical evidence of amyloidosis and in hospitalized non-Abyssinian cats. Affected and healthy Abyssinian cats, however, could not reliably be distinguished on the basis of SAA concentration, because of the wide range of SAA values in these 2 groups of cats.     

The contribution from hyphae, roots and organic carbon constituents to the aggregation of a sandy loam under long-term clover-based and grass pastures

Water-stable macro-aggregate size fractions (>2.0 mm, 1.0–2.0 mm, 0.5–1.0 mm and 0.25–0.5 mm) and non-aggregated soil from a sandy loam under long-term clover-based pasture and from grass pasture were analysed to determine the role of acid- and water-extractable carbohydrate C, total hyphal length, microbial biomass, organic C and total and mycorrhizal root length in stabilization of the aggregates. Aggregates were examined by scanning electron microscopy (SEM) and the particle-size distribution of the size fractions was also determined. Macro-aggregation increased under grass, relative to clover-based pasture; however, the properties of the aggregate fractions measured did not reflect this difference. Microbial-biomass C, extractable-carbohydrate C, hyphal length, total and mycorrhizal root length and organic C content of the soils were poorly correlated with macro-aggregation. Within the aggregates, the proportion of 250–1000-km sand was smaller and clay, silt and fine sand (20–250 μm) were greater relative to non-aggregated soil, suggesting that the >250-μm sand in the non-aggregated soil limited the stabilization of macro-aggregates. Under SEM, no enmeshment of aggregates by hyphae and roots was apparent. Although 50–160 m hyphae g^?1 soil was found within the aggregates, calculations showed that on average only 5 to 13 lengths of hyphae were associated with each 250-μm cube of soil within the aggregates, and suggested little potential to stabilize the aggregates by enmeshing. On average, all >2.0-mm aggregates contained less than 3.6 mm of roots and less than 50% by weight of <2.0-mm aggregates contained a single length of root. The findings cast doubt about the role of hyphae and fine roots in the stabilization of macro-aggregates through an enmeshing mechanism in sandy soils.     

Theileria parva: the immune status of calves born of dams immunised against East Coast fever

Eight Friesian cross cows three months pregnant to a single Friesian bull were immunised against East Coast fever by infection with Theileria parva (Muguga) sporozoite stabilate and treatment with pyrrolidino-methyl tetracycline. They were challenged with the homologous stock four times before calving and a fifth time after calving, and resisted all five challenges which killed all of the five groups of five susceptible controls. Calves born to these hyperimmunised dams were fully susceptible on challenge with the same stabilate, as were susceptible cows from the same farm and their calves. In both instances the calves died three to seven days earlier than the cows which were approximately 10 times heavier. These results show that one- to two-month-old taurine calves from artificially immunised dams are not protected from experimental T parva sporozoite challenge and that there is no inherent calfhood resistance to East Coast fever.     

Laryngeal paralysis in immature and mature dogs as one sign of a more diffuse polyneuropathy

Six dogs with laryngeal paralysis had clinical, electrophysiologic, and pathologic evidence of a more generalized polyneuropathy. Three of the dogs were young Dalmatians, one was a young Bouvier des Flandres, and two were older, large-breed dogs. The results of this study suggest that laryngeal paralysis in dogs may frequently be one clinical sign of an underlying, more generalized polyneuropathy. Two forms of this generalized polyneuropathy may exist: an early form, as seen in young dogs with congenital or hereditary disease, and a delayed-onset form that is usually found in older dogs with so-called idiopathic laryngeal paralysis, some of which may have hypothyroidism.     

Valvular pulmonic stenosis in bullmastiffs

Six bullmastiffs with congenital heart disease were evaluated over a three-year period. In all the cases the problem was shown to be dysplasia of the pulmonary valve. Five cases were male, one was female. All dogs available to follow-up developed signs of cardiac decompensation. Bullmastiffs must be added to the list of breeds in which valvular pulmonic stenosis is encountered frequently.     

The effect of exploratory laparotomy on the serum and peritoneal haptoglobin concentrations of the pony.

Serum haptoglobin concentration was used as an indicator of the acute phase response in ponies undergoing exploratory laparotomy. Preoperative, 1 h intraoperative, 3 h, 6 h, 12 h and 24 h postoperative blood samples and 48 h postoperative peritoneal fluid samples were obtained for haptoglobin analysis. A spectrophotometric assay based on cyanmethemoglobin binding capacity (CyanBC) was used to determine haptoglobin concentrations. The preoperative reference range for serum haptoglobin concentrations in these ponies was 25-60 mg CyanBC/dL. Intraoperative and 3 h postoperative blood samples had decreased haptoglobin concentrations when compared to preoperative values. Serum haptoglobin concentrations began to rise by the 6 h postoperative sample and were generally elevated above preoperative values by the 24 h postoperative sample. Two of the ten ponies had mild signs of postoperative colic which were associated with twofold elevations in serum haptoglobin concentrations and fivefold elevations in peritoneal fluid haptoglobin concentrations.