Male stimuli are necessary for female sexual behavior and uterine growth in prairie voles (Microtus ochrogaster)

In reproductively naive female prairie voles (Microtus ochrogaster) direct contact with male urine or housing in a male-soiled cage, in the absence of physical contact, resulted in increased uterine weights, but did not reliably elicit behavioral estrus (defined by lordosis). Physical contact with an unfamiliar male, for 1 hr or more, followed by 30 or 48 hr of continuous access to a male-soiled cage, induced lordosis in approximately two-thirds of the females tested. When females were physically exposed to a male for 18 hr and tested 6 hr later, 70% showed lordosis. However, when females receiving either 1 or 18 hr of male contact were removed from the presence of the male and placed in a clean cage for 24 hr, only 29-37% of the females subsequently showed lordosis. These results suggest that direct physical contact with the male or chemical stimuli from the male may be necessary to induce and maintain behavioral estrus in female prairie voles.     

Rapid preparation of fresh-frozen undecalcified bone for histological and histochemical analysis

An increasing need to be able to relate hard tissue morphology to its histochemistry, and the lack of an adequate methodology, prompted this investigation of polyvinylpyrrolidones of varying chain length as supporting films for the cryomicrotomy of fresh undecalcified cortical and trabecular bone. We developed a method that parallels histological procedures but is biochemically inert and functions at -25 degrees C. Using an LKB heavy-duty cryomicrotome, large numbers of serial sections suitable for histochemistry, immunocytochemistry, or rapid histology can be prepared with ease. Sections from mouse skull, ox vertebra, and human iliac crest illustrate the versatility of the method, and have been examined for the preservation of general morphology and of histochemical activity of those enzymes most prominent in remodeling. The procedure does not preclude the subsequent application of standard hard-tissue embedding methods to the remaining material. Comparison with plastic-embedded bone shows not only close similarities in the structural preservation of the frozen tissue, making possible histomorphometry, but also shows some interesting staining differences. These include the absence of differentiation of the calcification front with toluidine blue stain in fresh frozen specimens, and the presence in frozen osteoid tissue of specific methylene blue metachromasia missing from plastic preparations.     

Small rodents and other mammals associated with mountain meadows as reservoirs of Giardia spp. and Campylobacter spp

Sixty-five percent (469 of 722) of the fecal samples collected from small rodents in the central Washington Cascade mountains were positive for Giardia spp. Trapping studies showed that microtines of the genus Microtus were heavily infected with the parasite. Morphologically the cysts and trophozoites were of the Giardia duodenalis type. Small-rodent populations appear to maintain their infection throughout the year. Our data suggest that there is no difference in the percentage of positive animals in areas receiving a lot of human use as opposed to animals in those areas receiving very little or no human use. Giardia spp. were also found in elk and beaver fecal samples. Campylobacter spp. were recovered infrequently from the small rodents inhabiting alpine meadows. Of 551 specimens cultured, less than 1% were positive for the bacterium, and the isolates were identified as Campylobacter coli. Water voles were susceptible to a human isolate of Campylobacter jejuni and shed the bacterium for several weeks. C. jejuni was also isolated from a bear fecal sample collected from a protected watershed. Our studies indicate that microtines and possibly other small rodents inhabiting mountain meadows have a potential to act as a reservoir for both Giardia spp. and Campylobacter spp. Because these animals may carry human pathogens, they should be included in animal surveys designed to assess the health risks associated with mountain watersheds.     

Epitope and functional specificity of monoclonal antibodies to mouse interferon-gamma: the synthetic peptide approach

Spleen cells from hamsters immunized with recombinant mouse interferon-gamma (IFN-gamma) were fused with mouse myeloma cells, resulting in the production of four anti-IFN-gamma monoclonal antibodies. Binding of 125I-IFN-gamma by these protein A-bound antibodies was specifically blocked by cold IFN-gamma. Binding by three of these antibodies was also blocked by a synthetic peptide corresponding to the N-terminal 1-39 amino acids of IFN-gamma, whereas a corresponding C-terminal (95-133) peptide had no effect on binding. The N-terminal specificity of these three antibodies was confirmed by their specific binding of 125I-N-terminal (1-39) peptide. One of the N-terminal specific monoclonal antibodies inhibited both antiviral and macrophage priming (for tumor cell killing) activities of IFN-gamma, whereas the other two had no effect on either biologic function. The selectivity of the inhibition of IFN-gamma function was not due to a differential ability of the N-terminal specific antibodies to bind IFN-gamma. Blocking experiments with cold IFN-gamma and N-terminal peptide suggest that the epitope specificities of the monoclonal antibodies could be determined by the conformational or topographic structure of IFN-gamma. An exact determination of the epitope specificity of the monoclonal antibody that inhibited IFN-gamma function could provide insight into the structural basis for the role of the N-terminal domain in the biologic function of IFN-gamma. Polyclonal antibodies to either the N-terminal or the C-terminal peptides also inhibited both the antiviral and the macrophage-priming activities of IFN-gamma. All of the antibodies that inhibited IFN-gamma function also blocked binding of IFN-gamma to membrane receptor on cells, whereas antibodies that did not block function also did not inhibit binding. The data suggest that both the N-terminal and the C-terminal domains of IFN-gamma play an important role in its antiviral and macrophage-priming functions, possibly in a cooperative manner.     

Indirect evidence of impairment of platelet desaturase enzymes in diabetes mellitus

We measured the platelet total phospholipid fatty acid profiles of 20 insulin treated (Type I) diabetics, 20 non-insulin treated (Type II) diabetics and 20 matched non-diabetic controls to determine the relationship between the omega 6 and omega 3 series of fatty acids in diabetes. A significant inverse correlation between linoleic acid and arachidonic acid occurred in the normal subjects (r = -0.61; P less than 0.001) but was not seen in the Type I diabetics (r = -0.13; P = NS) or in the Type II diabetics (r = -0.27; P = NS). No significant correlation was seen between linolenic acid and eicosapentaenoic acid in the normal controls (r = -0.34; P = NS) or in the Type I diabetics (r = 0.21; P = NS) or in the Type II diabetics (r = -0.20; P = NS). The results suggest that a functional impairment of platelet delta 5 and delta 6 desaturase may occur in diabetes which disrupts the normal equilibrium between linoleic acid and arachidonic acid. However, the level of eicosapentaenoic acid appears to be less dependent on conversion from linolenic acid. Our findings are of importance to studies designed to reduce platelet aggregation in diabetics and non-diabetics by manipulation of the levels of the precursor fatty acids of thromboxane.     

The effects of certain vasodilating prostaglandins on the coronary and hindlimb vascular beds of the conscious sheep

Vasodilating prostaglandins were injected, in bolus doses, into the lower abdominal aorta or left circumflex coronary artery (LCCA) of conscious sheep. Local blood flow, mean arterial pressure (MAP), heart rate (HR) and ECG were monitored continuously. 6-Keto PGF1 alpha had no effect on either vascular bed in doses up to 100 micrograms. PGE2 was more potent than PGI2 in dilating hindlimb vasculature and PGE2 induced a more persistent hyperaemia whereas PGD2 elicited a biphasic response (constriction-dilation). PGE1, PGE2, PGD2 and PGI2 all produced dose-dependent vasodilation, the order of potency being PGD2 greater than PGI2 greater than PGE1 greater than or equal to PGE2. The effect of PGI2 was more transient and PGE1 and PGD2 caused small but consistent decreases in MAP and HR, respectively.     

Fluorescent color factor calculation using dBASE-II

A software system utilizing dBASE-II operating on a dual-driveApple II+ computer is described. Color factors and retentiontimes for 15 amino acids and -(-glutamyl)lysine dipeptide arecalculated following high performance liquid chromatography.The software package produces a listing of acceptable limitsfor these parameters calculated as plus and minus 2 standarddeviations of the mean. The code is distributed in source form. ; accepted on March 10, 1986     

Sexual differentiation of rat liver carbonic anhydrase III

Using radioimmunoassay, the concentration of carbonic anhydrase III in the livers of adult male rats was found to be approx. 30-times greater than that observed in mature females. Castration of male rats led to a marked reduction in liver carbonic anhydrase III concentrations which could be partially restored to control levels by testosterone replacement. Administration of testosterone to ovariectomised female rats induced about a 5-fold increase in liver carbonic anhydrase III concentration. Immunoprecipitation analysis of the products of liver mRNA translation in vitro with antiserum specific for carbonic anhydrase III showed that hormonal control of the levels of carbonic anhydrase III in liver is mediated by changes in the amount of translatable carbonic anhydrase III mRNA. Marked changes in liver carbonic anhydrase III concentrations were also observed in developing and ageing male rats.     

Frost injury and heterogeneous ice nucleation in leaves of tuber-bearing solanum species : ice nucleation activity of external source of nucleants

The heterogeneous ice nucleation characteristics and frost injury in supercooled leaves upon ice formation were studied in nonhardened and cold-hardened species and crosses of tuber-bearing Solanum. The ice nucleation activity of the leaves was low at temperatures just below 0°C and further decreased as a result of cold acclimation. In the absence of supercooling, the nonhardened and cold-hardened leaves tolerated extracellular freezing between −3.5° and −8.5°C. However, if ice initiation in the supercooled leaves occurred at any temperature below −2.6°C, the leaves were lethally injured.

To prevent supercooling in these leaves, various nucleants were tested for their ice nucleating ability. One% aqueous suspensions of fluorophlogopite and acetoacetanilide were found to be effective in ice nucleation of the Solanum leaves above −1°C. They had threshold temperatures of −0.7° and −0.8°C, respectively, for freezing in distilled H₂O. Although freezing could be initiated in the Solanum leaves above −1°C with both the nucleants, 1% aqueous fluorophlogopite suspension showed overall higher ice nucleation activity than acetoacetanilide and was nontoxic to the leaves. The cold-hardened leaves survived between −2.5° and −6.5° using 1% aqueous fluorophlogopite suspension as a nucleant. The killing temperatures in the cold-hardened leaves were similar to those determined using ice as a nucleant. However, in the nonhardened leaves, use of fluorophlogopite as a nucleant resulted in lethal injury at higher temperatures than those estimated using ice as a nucleant.

     

Biology of Aphidius rhopalosiphi, a parasitoid of cereal aphids

Summary Aphidius rhopalosiphi produced 212 offspring on average but in one case 509 deposited eggs were found. Superparasitization occurred but the frequency distribution of parasitoid larvae in aphids differed significantly from random, indicating a certain degree of host discrimination. Average life span of adults was 13.1 days and sex ratio was 1:1. It changed in time among successively produced offspring.Handling time was about 2 sec and was somewhat longer in the fourth instar than in the second. Second and third aphid instars were preferred for oviposition. Functional response was sigmoid and at an aphid density of 100 aphids per cage percentage parasitization decreased.

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Résumé Nous avons examiné quelques aspects des relations entre le parasitoïde Aphidius rhopalosiphi et son hôte Sitobion avenae. La fécondité potentielle à l'émergence (144 ufs) était inférieure à la fécondité réalisée: nombre de pucerons momifiés (212). La plupart des ufs étaient pondus dans la première semaine de la vie imaginale (fig. 1). Le super-parasi-tisme s'est produit dans les conditions expérimentales, mais la distribution des larves (fig. 2) s'écartait souvent de la distribution au hasard, ce qui indiquait une certaine discrimination parmi les hôtes.La longévité moyenne était de 13,1 jours et le maximum de 23 jours. Le taux sexuel (fig. 3) pour l'ensemble de la descendance était 1/1, mais changeait dans le temps. Après le premier jour de ponte, plus de femelles que de mâles étaient obtenues; mais après 6 jours, il n'y avait presque plus que des mâles.Le taux de rencontre des aphides et des parasitoïdes augmentait avec la taille (fig. 4a). Le temps de prospection de l'hôte était en moyenne de 2 secondes, mais dans certains cas il atteignait 10 secondes ou plus. La proportion de rencontres efficaces était plus faible au 4ème stade (fig. 4b). Il n'y avait pas de différence significative dans le nombre de larves de parasitoïdes obtenues par rencontre pendant les stades 1 à 3 des pucerons, mais elle était significativement inférieure au 4ème stade par rapport au second. Les taux globaux de succès (taux de rencontre et nombre de larves formées par rencontre) étaient plus élevés aux 2ème stades des pucerons. La réponse fonctionnelle était sigmoïde (fig. 5).