Sexual behavior of donkey jacks: influence of ejaculatory frequency and season

The courtship of 5 jacks was evaluated weekly for 12 mo. The characteristics of sexual behavior were recorded before the collection of 2 ejaculates, at 4-h intervals, into an artificial vagina in the presence of a female in either natural or induced estrus. The maximum time given to the jack to perform the ejaculatory mount was 1 h. If the jacks did not ejaculate, another attempt was made the following week. At the time of collection, the male and female remained free in a paddock (20 m2). The data presented in this study is based on results in which the first and second ejaculates occurred successively. Statistical analysis was done by a 2 x 2 factorial design in randomized blocks. The mean +/- SEM of sexual behavior characteristics for the first and second ejaculate were, respectively: time until first mount = 0.7 +/- 0.2 and 0.9 +/- 0.2 min; time until first erection = 11.9 +/- 1.1 and 11.6 +/- 1.1 min; time until ejaculation = 15.0 +/- 1.2 and 13.6 +/- 1.2 min; frequency of partial exposure of penis = 3.4 +/- 0.4 and 3.2 +/- 0.4; frequency of total exposure of penis = 1.0 + 0.2 and 1.1 +/- 0.2; frequency of flehmen responses = 6.6 +/- 0.5 and 4.6 +/-0.5; frequency of erections = 1.3 +/- 0.1 and 1.2 +/- 0.1; frequency of retreats away from the female = 1.8 +/- 0.2 and 1.0 +/- 0.2; frequency of mounts with erection but without ejaculation = 0.3 +/- 0.1 and 0.1 +/- 0.1; frequency of mounts without erection = 2.0 +/-0.1 and 1.2 +/- 0.1 ; and frequency of pelvic copulatory movements = 4.8 +/- 0.4 and 4.4 +/- 0.4. Individual differences were observed (P<0.05) for partial and total exposure, flehmen responses, mounts without erection and pelvic copulatory movements. The variables flehmen responses, retreats away from the female and mounts without erection showed significant differences (P<0.05) between ejaculates. Seasonal effects on sexual behavior characteristics were not found. However, a monthly effect was noted for flehmen responses, partial exposure of the penis, mounts without erection, retreats away from the female and pelvic copulatory movements when the 2 ejaculates were combined.     

Stimulation of the growth and solamargine production by Solanum paludosum multiple shoot cultures using a new culture medium

Organogenic callus cultures of Solanum paludosum were obtained from root, hypocotyle and cotyledon explants of plantlets cultured in sterile conditions. These callus cultures developed multiple shoots which proliferated in Murashige and Skoog basal liquid medium. These multiple shoots produced solamargine, the main steroidal glycoalkaloid present in the unripe fruits.The optimization of the macronutrient composition of the liquid medium was performed by a method derived from the plant composition. This approach results in the establishment of an appropriate medium (SPOM medium) suitable for the improvement of both growth and solamargine production by multiple shoot cultures of S. paludosum.     

Culture of rat mesenteric arteriolar smooth muscle cells: effects of platelet-derived growth factor, angiotensin, and nitric oxide on growth

We cultured smooth muscle cells as explants from rat mesenteric arterioles (40–200m in diameter) obtained by injecting a suspension of iron oxide intraarterially and magnetically separating the arterioles after collagenase digestion of adventitial tissue. In third-passaged cells we ascertained smooth muscle purity of >98% by characteristic morphology, contraction responses, and specific immunofluorescence staining. Treatment of growth-arrested (in 0.4% fetal calf serum) cells with platelet-derived growth factor (0.3–7.5 nM) or angiotensin II (0.001–1000 nM) induced ³H-thymidine incorporation and cell proliferation in a dose-dependent manner (P<0.01). S-nitroso-N acetylpencillamine (0.05–0.5 mM), a nitric oxide-generating compound, inhibited 10% fetal calf serum-induced ³H-thymidine incorporation (P<0.05) and cell proliferation (P<0.01). The antimitogenic effect of S-nitroso-N-acetylpencillamine was significantly reduced by hemoglobin and potentiated by superoxide dismutase (P<0.01). In addition to a new technique for culturing mesenteric arteriolar smooth muscle cells, these findings provide evidence that platelet-derived growth factor, angiotensin II, and nitric oxide may be involved in their growth control.     

A protocol for the efficient screening of putatively transformed plants forbar,the selectable marker gene,using the polymerase chain reaction

Amplification of thebar gene usingTaq DNA polymerase in PCR is often not successful, possibly due tobar's high GC content. We describe a PCR protocol in which reliable amplification at a sensitivity of one gene copy per genome (in this study, barley) present in the reaction was achieved using a novel pair of primers and Expand^tm High Fidelity DNA polymerase mix (Boehringer Mannheim). This method should allow for rapid screening of plants putatively transformed withbar.     

Measures of geographic range size: the effects of sample size

A number of methods have been used for quantifying the sizes of the geographic ranges of species. The consequences of different levels of sampling (the proportion of actual spatial occurrences) are explored for eight of these, using data on the occurrences of butterfly species on a 10 × 10 km grid across Britain. For all methods, the percentage error of estimation (PEE) decreases with the number of 10 × 10 km squares which a species occupies, most rapidly for extent measures, and more rapidly for area measures than for measures of numbers of units occupied. The rate of decline in PEE itself falls as sampling effort increases. At a given sampling level, rank correlations between range sizes measured by different methods are generally high, but there is no consistent change in the magnitude of these correlations as the level of sampling increases. The composition of the set of species with the smallest range sizes changes with the level of sampling.     

The 5'' end of yeast 5.8S rRNA is generated by exonucleases from an upstream cleavage site.

We have developed techniques for the detailed analysis of cis-acting sequences in the pre-rRNA of Saccharomyces cerevisiae and used these to study the processing of internal transcribed spacer 1 (ITS1) leading to the synthesis of 5.8S rRNA. As is the case for many eukaryotes, the 5' end of yeast 5.8S rRNA is heterogeneous; we designate the major, short form 5.8S(S), and the minor form (which is seven or eight nucleotides longer) 5.8S(L). These RNAs do not have a precursor/product relationship, but result from the use of alternative processing pathways. In the major pathway, a previously unidentified processing site in ITS1, designated A3, is cleaved. A 10 nucleotide deletion at site A3 strongly inhibits processing of A3 and the synthesis of 5.8S(S); processing is predominantly transferred to the alternative 5.8S(L) pathway. Site A3 lies 76 nucleotides 5' to the end of 5.8S(S), and acts as an entry site for 5'-->3' exonuclease digestion which generates the 5' end of 5.8S(S). This pathway is inhibited in strains mutant for XRN1p and RAT1p. Both of these proteins have been reported to have 5'-->3' exonuclease activity in vitro. Formation of 5.8S(L) is increased by mutations at A3 in cis or in RAT1p and XRN1p in trans, and is kinetically faster than 5.8S(S) synthesis.     

Immunochemical and immunohistological expression of Lewis histo-blood group antigens in small intestine including individuals of the Le(a+b+) and Le(a−b−) nonsecretor phenotypes

Histological samples and total non-acid glycosphingolipids were prepared from small intestine of human cadavers with the Le(a+b+) and Le(a–b–) nonsecretor phenotypes and contrasted with the more common Lewis phenotypes. Glycolipid fractions were analysed by thin-layer chromatography and tested for Lewis activity with monoclonal antibodies reactive to Lewis epitopes. Paraffin-embedded small intestine sections were also fluorescently immunostained with anti-Lewis antibodies. Unlike the common Lewis positive phenotypes, we were immunochemically able to demonstrate the copresence of large amounts of Le^a and Le^b glycolipids in the Le(a+b+) sample. In addition we demonstrated increased formation of extended Lewis structures in this phenotype. By immunohistochemistry Le^a, Le^b and type 1 precursor chain epitopes could be demonstrated in the brush border. These results show that the expression of the Le(a+b+) phenotype at the erythrocyte phenotyping level parallels the small intestinal expression of this phenotype, and the patterns of Lewis antigen expressions are unique to this phenotype. By immunohistochemistry and immunochemistry we also demonstrated the presence of trace amounts of Lewis active glycoconjugates in the small intestine of the Le(a–b–) nonsecretor and Le(a+b–) samples. In the Le(a–b–) nonsecretor Le^a and Le^b activity was absent and type 1 precursor was present in brush border, while Le^b activity was immunohistologically demonstrated in the Golgi apparatus of the deep glands. Trace amounts of both Le^a and Le^b glycolipids were identified in this sample. In parallel trace Le^b activity could also be detected in the glycolipids of the Le(a+b–) sample and could be immunohistologically demonstrated to be fully expressed in occasional cells in the deep glands of the small intestine, a pattern quite dissimilar to that of the Le(a–b–) nonsecretor. The results in this paper show that the expression of Lewis glycoconjugates in the small intestine parallel the expression of Lewis erythrocyte phenotypes. However, inappropriate Lewis activity is also seen in individuals of other phenotypes and the mechanisms by which these Lewis antigens are made appears to be different for different phenotypes.Abbreviations FITC fluorescein isothiocyanate - HPLC high-performance liquid chromatography - NeuAc N-acetyl-d-neuraminic acid - RBC red blood cell - TLC thin-layer chromatography - TRITC tetramethyl rhodamine isothiocyanate     

Expression of Lewis histo-blood group glycolipids in the plasma of individuals of Le(a+b+) and partial secretor phenotypes

Red cell Lewis antigens are carried by glycosphingolipids passively absorbed from plasma. Plasma was collected from a spectrum of individuals with normal and unusual Lewis/secretor phenotypes in order to investigate the glycolipid basis for the unusual phenotypes. Samples were obtained from: a Le(a+b–) ABH nonsecretor who secreted Lewis substances; a Le(a+b–) partial secretor; Le(a+b+) partial secretors; Le(a+b+) secretors; and a full range of normal Lewis/secretor phenotypes as controls. The Le(a+b+) samples represented Polynesian, Asian and Réunion Island ethnic backgrounds. Nonacid glycolipids were prepared, separated by thin-layer chromatography, and then immunostained with potent monoclonal antibodies of known specificity. Despite different serological profiles of the Le(a+b–) and Le(a+b+) Polynesian samples, their plasma glycolipid expressions were very similar, with both Le^a and Le^b co-expressed. The copresence of Le^a and Le^b in Le(a+b+) samples is in marked contrast to Caucasians with normal Lewis phenotypes, who have predominantly either Le^a or Le^b. These results suggest that there is a range of the secretor transferases in different individuals, possibly due to different penetrance or to several weak variants. We also show that Lewis epitopes on longer and/or more complex core chains appear to be predominant in the Polynesian Le(a+b+) samples. The formation of these extended glycolipids is compatible with the concept that in the presence of reduced secretor fucosyltransferase activity, increased elongation of the precursor chain occurs, which supports the postulate that fucosylation of the precursor prevents or at least markedly reduces chain elongation.Abbreviations CBA chromatogram binding assay - TLC thin-layer chromatography