Perfusion method for preparing pig brain cortex for Golgi-Cox impregnation

The perfusion procedure described in this paper produces high quality impregnation of pig visual and somatosensory cortical neurons with a Golgi-Cox solution. Starting within 30 min after death, pig heads were perfused with a fixative solution composed of a mixture (v/v) of liquid phenol, 5%; formalin, 14%; ethylene glycol, 25%; methanol, 28%; and water, 28% for two periods of 4 hr each. After perfusion, the heads were chilled for at least 18 hr. The entire brain was removed from the skull and then placed in 10% buffered formalin, where it remained for at least 10 days before taking the blocks that were to be immersed in the Golgi-Cox solution. Three weeks spent in the Golgi-Cox solution typically produced uniform neuron impregnation. The tissue blocks were then embedded in celloidin and sectioned at 120 micron. This procedure avoids the following difficulties: Golgi-Cox methods that produced excellent results with rodent or primate tissue were unsuccessful with pig tissue, placing fresh tissue in Golgi-Cox solution resulted in incomplete neuron impregnation, and immersion fixation in 10% buffered formalin without perfusion resulted in excessive staining of glia.     

Studies of the seminal receptacle of the crab Portunus sanguinolentus (Portunidae: Brachyura)

The seminal receptacle or spermatheca of Portunus sanguinolentus consists of two parts--an anterior glandular and a posterior chitinous part. The chitinous part continues as the oviduct, which opens on the sternite of the sixth thoracic segment. Significant morphological and histological differences were observed between the spermatheca, as well as the oviduct, of mated and unmated crabs. In mated crabs the spermatheca is much more bulging, owing to receipt of a copious supply of seminal products, and its cells are hyperactive. Further stages of ovarian development were observed as indicators of sequential changes in the spermatheca. The secretory cells gradually disintegrate by way of holocrine secretion; this results in cellular stratification and the formation of distinct furrows in the chitinous posterior part.     

Effects of temperature and infection with Eimeria ochrogasteri on digestive organs of the prairie vole, Microtus ochrogaster

Prairie voles, Microtus ochrogaster, were infected with Eimeria ochrogasteri and exposed to 2 environmental temperatures, 5 and 22 C. Dry weights of the small and large intestines increased by 33% and 19%, respectively, in infected animals. Infected animals also exhibited a 14% decrease in cecal length compared to uninfected animals. The interaction between temperature and infection affected the length of the small intestine. Infected animals maintained at 5 C had longer small intestines than both infected animals housed at 22 C, and uninfected animals at 22 or 5 C. Furthermore, the dry weight of the small intestine was affected by a 3-way interaction (infection, temperature, and sex). Temperature affected stomach and liver dry weights, as well as lengths of the small intestine and cecum. Stomach and liver dry weights, as well as small intestine lengths, were greater in those animals held at 5 C, whereas cecum lengths decreased. Prepatency, patency, and total oocyst production were not affected by temperature; however, infected animals held at 5 C exhibited diarrhea during the patent period.     

Parallel dose-response curves in combination experiments

A possible experimental design for combination experiments is to compare the doseresponse curve of a single agent with the corresponding curve of the same agent using either a fixed amount of a second one or a fixed dose ratio. No interaction is then often defined by a parallel shift of these curves. We have performed a systematic study for various types of doseresponse relations both for the dose-additivity (Loewe additivity) and for the independence (Bliss independence) criteria for defining zero interaction. Parallelism between doseresponse curves of a single agent and those of the same agent in the presence of a fixed amount of another one is found for the Loewe-additivity criterion for linear doseresponse relations. For nonlinear relations, one has to differentiate between effect parallelism (parallel shift on the effect scale) and dose parallelism (parallel shift on the dose scale). In the case of Loewe additivity, zero-interaction dose parallelism is found for power, Weibull, median-effect and logistic doseresponse relations, given that special parameter relationships are fulfilled. The mechanistic model of competitive interaction exhibits dose parallelism but not effect parallelism for Loewe additivity. Bliss independence and Loewe additivity lead to identical results for exponential doseresponse curves. This is the only case for which dose parallelism was found for Bliss independence. Parallelism between single-agent doseresponse relations and Loewe additivity mixture relations is found for examples with a fixed doseratio design. However, this is again not a general property of the design adopted but holds only if special conditions are fulfilled. The comparison of combination doseresponse curves with single-agent relations has to be performed taking into account both potency and shape parameters. The results of this analysis lead to the conclusion that parallelism between zero interaction combination and single-agent doseresponse relations is found only for special cases and cannot be used as a general criterion for defining zero-interaction in combined-action assessment even if the correct potency shift is taken into account.     

Insulin Sensitivity of Heifers on Different Diets

The hyperinsulinemic euglycemic clamp technique was used to investigate the effect on insulin sensitivity of 2 different diets used in practical cattle feeding in calves. Ten 4 to 5-month-old heifer calves were allocated to 2 feeding groups, LO or HI, to obtain growth rates of 400 g/day or 900 g/day. The heifers were fed and housed individually for 5 weeks. Growth rates close to calculated rates were obtained with the diets used. Weekly blood samples were collected from the jugular vein for analysis of glucose, insulin, cortisol, total serum protein, urea, cholesterol and nonesterified fatty acids. During week 5, insulin sensitivity was estimated using the hyperinsulinemic euglycemic clamp technique. Insulin sensitivity did not differ between the groups, but the plasma glucose levels were higher during weeks 3 and 4 for the HI group compared to the LO group. It may be concluded that the amount of concentrate in the diet was too low to induce changes in either the basal plasma insulin levels or the insulin sensitivity in the HI group.     

Dietary restriction causes chronic elevation of corticosterone and enhances stress response in red-legged kittiwake chicks

Release of corticosterone in hungry kittiwake chicks facilitates begging and allows them to restore depleted energy reserves by increasing parental food provisioning. However, in order to avoid detrimental effects of chronic elevation of corticosterone, chicks might suppress adrenocortical activity in response to prolonged food shortages. In this study we examined temporal dynamics of corticosterone release in red-legged kittiwake (Rissa brevirostris) chicks exposed to prolonged restrictions in energy content and/or nutritional quality (low versus high lipid content) of their food. Starting at the age of 15 days, chicks were fed either high- or low-lipid fish at 40%, 65%, and 100% of ad libitum energy intake. Body mass measurements and baseline plasma samples were taken on a weekly basis after beginning of the treatment. After 3 weeks of treatment, chicks were exposed to a standardized acute handling and restraint stress protocol, where in addition to a baseline sample, three plasma samples were taken at intervals up to 50 min. We found that food-restricted chicks had lower body mass, chronically (during 2-3 weeks) elevated baseline and higher acute stress-induced levels of corticosterone compared to chicks fed ad libitum. Low lipid content of food further exacerbated these effects. An increase in baseline levels of corticosterone was observed within a week after energy requirements of food-restricted chicks exceeded their daily energy intake. A tendency for suppression of adrenocortical activity was observed in treatments fed low-lipid diets only at the end of the experiment. We suggest that nest-bound chicks, if food-stressed, might suffer deleterious effects of chronic elevation of corticosterone.