Electrophoretic map of boar sperm plasma membrane polypeptides and localization and fractionation of specific polypeptide subclasses

A high resolution, two-dimensional (2-D) electrophoretic map of the plasma membrane (PM) polypeptides from the ejaculated boar spermatozoon is described. 2-D silver-stained polyacrylamide gel electrophoresis (PAGE) gels revealed over 250 polypeptides; Coomassie blue staining revealed more than 100. Fifty Coomassie-staining polypeptides were catalogued and biochemically characterized, with twenty of these designated major sperm PM polypeptides. Cavitation pressures ranging from 50 PSI to 1000 PSI were used to enrich 2-D maps either in head PM (50 PSI) or in flagellar PM (1000 PSI) and provided tentative localization of certain PM polypeptides. Immunoabsorption chromatography showed that most major polypeptides seen in the 2-D map were antigenic. Major polypeptide bands from single dimensional (1-D) gels were excised, antibodies against them were produced in rabbits, and the polypeptides were localized via indirect fluorescein isothiocyanate (FITC) technique. Cross-reacting antigenic determinants in the PAGE PM polypeptides were determined by transblotting and staining the transblots by an indirect peroxidase technique. Cross-reactivity was extensive with several polypeptide groups, but specific enough with others to allow tentative localization. Lectin affinity chromatography using Con A, WGA, RCA-1, PNA, and DBA indicated the lectin specificity of PM polypeptides. These data together with periodic acid-Schiff (PAS) and carbohydrate-specific silver staining permitted identification of glycoproteins in the 2-D maps. FITC coupled to specific lectins showed the regional distribution of these lectins on the sperm surface. The 2-D polypeptide map and the catalogue of properties of major Coomassie-stained PM polypeptides provides a reference for future studies in the boar and other species.     

Fermentation of Peptides by Bacteroides ruminicola B14

The maximum growth rate of Bacteroides ruminicola B₁4 was significantly improved when either Trypticase or acetate and C₄-C₅ fatty acids were added to defined medium containing macrominerals, microminerals, vitamins, hemin, cysteine hydrochloride, and glucose. The organism was unable to grow with peptides as the sole energy source, but growth yields from glucose were significantly improved when Trypticase was added to batch cultures containing basal medium, acetate, and C₄-C₅ volatile fatty acids. During periods of rapid growth, very little peptide was deaminated to ammonia, but after growth ceased there was a linear increase in ammonia. Fifteen grams of Trypticase per liter resulted in maximum ammonia production. In glucose-limited chemostats, ammonia production from peptides was inversely proportional to the dilution rate, and 87% of the variation in ammonia production could be explained by retention time in the culture vessel. Chemostats receiving Trypticase had higher theoretical maximum growth yields and lower maintenance energy expenditures than similar cultures not receiving peptide. Cells from the Trypticase cultures contained more carbohydrate, and this difference was most evident at rapid dilution rates. When corrections were made for cell composition and the amount of peptides that were fermented, it appeared that peptide carbon skeletons could be used for maintenance energy. B. ruminicola B₁4 was unable to grow on peptides alone because it was unable to utilize peptides at a fast enough rate to meet its maintenance requirement.     

Generalized Transduction in the Phytopathogen Pseudomonas syringae

Bacteriophages isolated from culture supernatants of Pseudomonas syringae pv. syringae and from sewage transferred various chromosomal genes to P. syringae PS224. Linkage between arginine and tryptophan loci was demonstrated. The number of transductants recovered per milliliter was not altered appreciably by UV irradiation of selected phage isolates. In addition, the presence of the IncP2 plasmid R38 in a P. syringae PS224 arginine auxotroph did not increase the transduction frequency as it does in Pseudomonas aeruginosa. Increasing the multiplicity of infection of transducing phage Pssy15 from 1 to 10 resulted in up to a 10-fold increase in the number of transductants recovered, although the actual transductional frequency remained about the same. Treatment of transduction mixtures with DNase did not affect transductional frequency.     

Ribosomal genes of Neurospora crassa: constancy of gene number in the conidial and mycelial phases, and homogeneity in length and restriction enzyme cleavage sites within strains

We report the results of experiments which, while not specifically designed to study the possibility of rDNA amplification during different developmental stages in the N. crassa life cycle, clearly indicate a relative constancy in the rDNA content of conidia (asexual spores) and mycelial cells. We also report the results of restriction enzyme studies which indicate that the Neurospora rDNA repeat units are homogeneous in length and restriction site pattern within any given Neurospora strain. These results directly contradict the recent report of Dutta et al. (1983), in which the authors concluded that the rDNA of germinating conidia is amplified, relative to mycelia, and that up to 10% of the rDNA units are heterogeneous.     

Relationship of seminal vesicle size and measures of libido in yearling beef bulls

Ninety-five yearling beef bulls were given routine Breeding Soundness Examinations, two libido tests and palpated rectally for internal genital disease and measurement of seminal vesicle size. Finger tips were premeasured, and then used as "glandometers". The bulls were examined at the end of a 140-day performance test. Sixteen lines of breeding were examined including 13 Hereford, 2 Angus and 1 Red Angus. Average age was 384 days and average weight was 961 lbs. Line and breed differences (P<.05) were observed for scrotal circumference, scrotal circumference score and second libido test. Similar differences were observed for SV length (P<.01), depth and volume both (P<.05). There were no significant correlations between seminal vesicle (SV) size and libido scores. There were, however, significant correlations between SV size and scrotal circumference (P<.05), BSE score (P<.01), body weight (P<.01), and semen morphology score (P<.05). Proximal droplets and midpiece abnormalities, respectively, were the most common spermatozoal abnormalities observed in semen from this group of bulls.     

Effects of deficiencies in the engrailed region of Drosophila melanogaster

The engrailed gene of Drosophila melanogaster is believed to be involved in control of determination and differentiation of posterior compartments. en1/en1 causes a partial transformation of the posterior compartment of wing and first leg to mirror-image anterior, which prompted the hypothesis that engrailed + is a "selector gene" required for the posterior pathway decision. The incomplete transformation was thought due to residual en+ activity in en1; a deletion of engrailed (en28) was constructed to determine if a complete transformation can occur. en28 is homozygous lethal and cell lethal. en28/en1 survives to adult stage, but causes a weaker transformation than en1/en1, indicating that en1 is not a simple hypomorph. A more distal deletion, en30, survives over en-lethal alleles. Both en30/en1 and en28/en30 survive to adult stage, but do not cause a stronger posterior to anterior transformation than en1/en1; thus this effect may be allele specific. New abnormalities included (1) transformation of the posterior wing blade to haltere, an effect dependent on the bx+ (but not pbx+) pseudoallele of the bithorax complex; (2) abnormal bristle pattern, tarsal fusion, and degenerate posterior claws of all legs. Although these abnormalities are posterior compartment specific, they are not expected of a "selector gene." Thus the function of engrailed may be more complex than originally believed.     

Effect of Glutaraldehyde on the Outer Layers of Escherichia coli

S ummary : Sodium lauryl sulphate (SLS) at pH 3 and 8 lysed cell walls of Escherichia coli. Pretreatment with glutaraldehyde at pH 3 and at pH 8 prevented this lysis. SLS induced maximum lysis of E. coli cells at 40°; pretreatment of cells with glutaraldehyde prevented this lysis also. Electrophoretic studies indicated that glutaraldehyde accumu lated on the surface of E. coli cells more rapidly in acid than in alkaline conditions, and that it blocked amino groups on the surface layer of Bacillus subtilis spores. The relationship of these findings to the bactericidal efficiency of glutaraldehyde in acid and alkaline solution is discussed.     

Quantitative Nasal Culture: a Tool in Antibiotic Research

The use of the quantitative nasal culture was investigated as a means of evaluation of new antimicrobial drugs in man. Cyclacillin was somewhat more active in vitro than penicillin G against penicillin G-resistant organisms. Cyclacillin was highly effective in suppressing staphylococci susceptible to penicillin G in nasal carriers but did not suppress staphylococci resistant to penicillin G. Although in previous studies by others cyclacillin was effective in treating mice infected with penicillin G-resistant staphylococci, in the present studies cyclacillin was not effective in suppressing nasal penicillin G-resistant staphylococci in man at doses which markedly suppressed penicillin G-sensitive organisms.     

Immunological and Biophysical Separation of Dengue-2 Antigens

Antigenic compositions of slowly sedimenting dengue-2 hemagglutinin (SHA) and soluble complement-fixing antigen (SCF) were compared with the virion (rapidly sedimenting hemagglutinin, RHA) by radioimmune precipitation (RIP), RIP inhibition, kinetic neutralization, and neutralization blocking tests with the use of hyperimmune mouse ascitic fluids. RHA and SHA were unable to inhibit completely the RIP of each other by anti-RHA, and neutralization by anti-RHA was not blocked by SHA. This indicated that SHA is serologically related, but not identical, to RHA. SHA differed from RHA in that SHA lacked the “core” polypeptide but contained the two envelope polypeptides. In addition, SHA contained a polypeptide with a molecular weight of 16,500 daltons and a suggestion of several other proteins. These data, when considered with other evidence, suggest that SHA is a special form of “incomplete virus.” SCF was unable to inhibit the RIP of SHA or RHA or to block neutralizing antibodies. Further, anti-SCF did not neutralize RHA or precipitate significant levels of SHA or RHA. Polyacrylamide gel electrophoresis separated SCF from structural polypeptides by molecular size. This evidence suggests that SCF is a nonstructural antigen.