Functional parameters before and after parathyroidectomy: a prospective, randomized long-term trial on different rat strains.

For clinical controls before and after parathyroidectomy and for evaluation of the function of transplants of parathyroid tissue, it is necessary to establish standard values of relevant laboratory parameters for donor and recipient animals as well as for different types of nutrition. Since no such data are yet available, it was the purpose to define such standards. In a prospective randomized trial on 400 rats of the Dark Agouti (DA) and Lewis strain, different functional laboratory parameters such as total calcium, intact parathyroid hormone, phosphate, 1.25-dihydroxyvitamin D, and alkaline phosphatase were measured under a standard and low calcium diet over a period of 40 weeks. Two hundred of these animals underwent a parathyroidectomy four weeks after the beginning of the study and specimens were evaluated histologically. For all eight different study groups normal values could be defined within tight limits for parameters which describe the function of the parathyroid gland or elements of calcium metabolism under different conditions. The optimal conditions for a transplantation model of parathyroid glands were established. Lewis-rats were identified as the ideal donor and DA rats as the better recipient animals. These data can serve as reference values for future studies on transplantation of the parathyroid without immunosuppression.     

Evidence for vitamin D-independent active calcium absorption in newborn piglets

Summary The role of 1,25-dihydroxycholecalciferol (calcitriol) for intestinal calcium (Ca²⁺) absorption was studied in newborn (<1 week old) and weaned piglets (>6 weeks old). In both groups, normal piglets and piglets suffering from inherited pseudo vitamin D-deficiency rickets, type I (PVDRI) were used. In this inherited disorder, renal production of calcitriol is absent. Plasma samples were assayed for calcitriol and total Ca, and dissociation constants (K_d) and maximum binding capacities (B_max) of intestinal calcitriol receptors were determined under equilibrium conditions at 4°C. Unidirectional Ca²⁺-flux rates were measured across stripped duodenal mucosae in Ussing chambers in the absence of electrochemical gradients. The plasma calcitriol concentrations of neonatal (26.5±7.1 pg/ml, n=11; ± SEM) and weaned PVDRI piglets (18.8±5.7 pg/ml, n=8)were unphysiologically low and differed significantly from control animals (83.6±14.8 pg/ml, n=8, and 86.9±9.6 pg/ml, n=11, respectively). However, newborn PVDRI piglets had normal plasma Ca levels at least during the first days of life. They became hypocalcemic and developed clinical symptoms of rickets during the following weeks. In newborn PVDRI and control piglets, B_max was significantly lower (84±28 fmol/mg protein and 127±55 fmol/mg protein, n=9, respectively) than in weaned piglets (741±82 fmol/mg protein, n=9, and 778±121 fmol/mg protein, n=8, respectively). Significant net Ca²⁺-fluxes were found in both newborn PVDRI and control piglets (88.8±25.1 nmol · cm^-2 · h^-1, n=6, and 86.5±10.5 nmol · cm⁻² · h⁻¹,n=9, respectively). However, active net Ca²⁺ absorption was completely absent in weaned PVDRI piglets. These results indicate the presence of vitamin D-independent mechanisms for active intestinal Ca²⁺ absorption during early postnatal life in pigs.     

Disappearance from blood and urinary excretion of 35S-thiamin in sheep. A kinetic study.

In seven experiments with two adult sheep a single dose of 35S-thiamin was injected intravenously. Blood and urine samples were taken at short intervals after dosing and analyzed for 35S-radioactivity. The data obtained were subjected to a curve fitting computer program. The disappearance of 35S-thiamin from blood followed a three-exponential function; the average half-times of the three components were 0.4, 3.7 and 93.7 minutes. Urinary excretion of the tracer was also three-exponential and amounted to 19--32% of the injected dose over 150 minutes.     

The passage of protozoa from the reticulo-rumen through the omasum of sheep

1. Protozoa in rumen contents and omasal effluent of growing wethers were counted. The wethers were equipped with rumen and abomasal cannulas, and omasal sleeves attached to the omasal-abomasal orifice. Rumen fluid dilution rates were elevated by continuous infusions of hypertonic mineral solutions (3-4 litres/d) for 24 d. Rumen contents and omasal effluent were sampled between 9 and 21 h during the last 10 d of each experiment. 2. Protozoal concentrations in omasal effluent were only 0.2-0.3 those found in the rumen under normal conditions. The ratio of protozoal concentrations in rumen: those in omasal effluent was for small Diplodinium spp. 4.6 (SD 0.9), for Ophryoscolex spp. 4.3 (SD 1.0), for Dasytricha ruminantium 4.0 (SD 0.5), for Isotricha spp. 3.8 (SD 0.8), for Entodinium spp. 3.6 (SD 0.9) and for Polyplastron multivesiculatum 2.6 (SD 0.5). 3. Elevation of rumen fluid dilution rate by 20 and 55% respectively, increased protozoal concentrations in omasal effluents from 22 to 33% and from 31 to 47% those in rumen contents. The apparent residence times of protozoa in the rumen were decreased 50% by the infusion of a mineral-salt solution. The increase in rumen fluid dilution rate had no significant effect on concentrations of protozoa in the rumen or on the differences of the apparent residence times between different species. The apparent residence time of holotrichs remained the same before and after infusion of the mineral-salt solution. 4. Apparent residence times of individual species of protozoa in the rumen were, under normal feeding conditions, 2.55 d, and were four to six times longer than the mean residence time of CrEDTA in the rumen.     

Studies on the toxicity of deoxynivalenol (DON), sodium metabisulfite,DON-sulfonate (DONS) and de-epoxy-DON for porcine peripheral blood mononuclear cells and the Intestinal Porcine Epithelial Cell lines IPEC-1 and IPEC-J2, and on effects of DON and DONS on piglets

The in vitro effects of deoxynivalenol (DON), de-epoxy-DON, DON-sulfonate (DONS) and sodium metabisulfite (Na₂S₂O₅, SBS) on porcine peripheral blood mononuclear cells (PBMC), and on the Intestinal Porcine Epithelial Cell lines IPEC-1 and IPEC-J2 were examined by using the MTT assay.In addition, an uncontaminated and a DON contaminated triticale were included in diets either untreated (CON, FUS) or SBS treated (CON-SBS, FUS-SBS) and fed to piglets for 28 d starting from weaning. The diet concentrations of DON and DONS amounted to 0.156, 2.312, 0.084 and 0.275 mg and to <0.05, <0.05, <0.05 and 1.841 mg/kg, respectively. PBMC of the so-exposed piglets were also subjected to the MTT assay.Neither DONS and SBS nor de-epoxy-DON affected the viability of PBMC, IPEC-1 and IPEC-J2 significantly up to concentrations of 17, 8 and 23 μM, respectively. For DON, IC₅₀ values were estimated at 1.2 ± 0.1, 1.3 ± 0.5 and 3.0 ± 0.8 μM for PBMC, IPEC-1 and IPEC-J2, respectively.PBMC from piglets fed the SBS treated diets were characterized by a significantly decreased stimulation index and an increased IgA supernatant concentration with the SBS effect being significantly more pronounced after feeding the FUS-SBS diet. Further studies should clarify the possible impact of SBS on the porcine immune system.     

Does Dietary Deoxynivalenol Modulate the Acute Phase Reaction in Endotoxaemic Pigs?—Lessons from Clinical Signs,White Blood Cell Counts,and TNF-Alpha

We studied the interaction between deoxynivalenol (DON)-feeding and a subsequent pre- and post-hepatic immune stimulus with the hypothesis that the liver differently mediates the acute phase reaction (APR) in pigs. Barrows (n = 44) were divided into a DON-(4.59 mg DON/kg feed) and a control-diet group, surgically equipped with permanent catheters pre- (V. portae hepatis) and post-hepatic (V. jugularis interna) and infused either with 0.9% NaCl or LPS (7.5 µg/kg BW). Thus, combination of diet (CON vs. DON) and infusion (CON vs. LPS, jugular vs. portal) created six groups: CON_CON_jug.-CON_por., CON_CON_jug.-LPS_por., CON_LPS_jug.-CON_por., DON_CON_jug.-CON_por., DON_CON_jug.-LPS_por., DON_LPS_jug.-CON_por.. Blood samples were taken at −30, 15, 30, 45, 60, 75, 90, 120, 150, 180 min relative to infusion and analyzed for leukocytes and TNF-alpha. Concurrently, clinical signs were scored and body temperature measured during the same period. LPS as such induced a dramatic rise in TNF-alpha (p < 0.001), hyperthermia (p < 0.01), and severe leukopenia (p < 0.001). In CON-fed pigs, an earlier return to physiological base levels was observed for the clinical complex, starting at 120 min post infusionem (p < 0.05) and persisting until 180 min. DON_LPS_jug.-CON_por. resulted in a lower temperature rise (p = 0.08) compared to CON_LPS_jug.-CON_por.. In conclusion, APR resulting from a post-hepatic immune stimulus was altered by chronic DON-feeding.     

Minimally invasive closed-chest ultrasound-guided substance delivery into the pericardial space in mice

Organ-directed gene transfer remains an attractive method for both gaining a better understanding of heart disease and for cardiac therapy. However, virally mediated transfer of gene products into cardiac cells requires prolonged exposure of the myocardium to the viral substrate. Pericardial injection of viral vectors has been proposed and used with some success to achieve myocardial transfection and may be a suitable approach for transfection of atrial myocardium. Indeed, such an organ-specific method would be particularly useful to reverse phenotypes in young and adult genetically altered murine models of cardiac disease. We therefore sought to develop a minimally invasive technique for pericardial injection of substances in mice. Pericardial access in anaesthetised, spontaneously breathing mice was achieved using continuous high-resolution ultrasound guidance. We could demonstrate adequate delivery of injected substances into the murine pericardium. Atrial epicardial and myocardial cells were transfected in approximately one third of mice injected with enhanced green fluorescent protein-expressing adenovirus. Cellular expression rates within individual murine atria were limited to a maximum of 20 %; therefore, expression efficiency needs to be further improved. Minimally invasive, ultrasound-guided injection of viral material appears a technically challenging yet feasible method for selective transfection of atrial epi- and myocardium. This pericardial injection method may be useful in the evaluation of potential genetic interventions aimed at rescuing atrial phenotypes in transgenic mouse models.