Epitope analysis of a sperm acrosomal antigen defined by HSA-5 monoclonal antibody

Among the monoclonal antibodies recommended by the WHO Sperm Antigen Workshop for immunocontraceptive vaccine development, HSA-5 showed a high degree of sperm specificity and significantly inhibited in vitro fertilization in both humans and mice. Using a Western blot assay, HSA-5 was found to recognize a sperm antigen designated as HSAg-5 (human) or MSAg-5 (mouse) which ranged in molecular weight from 18 to 100 kDa. This monoclonal antibody was used as the probe for the immunoscreening of mouse testis cDNA libraries constructed in the lambda gt-11 expression vector. One of the positive cDNA clones was shown to have a cDNA insert of approximately 1 kb and to encode a recombinant fusion protein containing 77 amino acid residues in the C-terminal region of MSAg-5. This 1 kb cDNA insert was engineered in a pGEX vector to express a recombinant glutathione S-transferase fusion protein (GST-5). Using an enzyme-linked immunosorbent assay (ELISA) and Western blot analysis, both anti-GST-5 sera and the monoclonal antibody were shown to react with GST-5. The Northern blot of a mouse testis RNA preparation revealed that the isolated cDNA probe hybridized with a 4.0 kb mRNA. Several oligopeptides were synthesized based on the predicted C-terminal hydrophilic regions of the recombinant fusion protein. Using ELISA and a dot blot assay, peptide regions containing the immunogenic epitopes recognized by HSA-5 monoclonal antibody were identified.     

Improving vineyard sampling efficiency via dynamic spatially explicit optimisation

Background and Aims: Environmental variables within vineyards are spatially correlated, impacting the economic efficiency of cultural practices and accuracy of viticultural studies that utilise random sampling. This study aimed to test the performance of non‐random sampling protocols that account for known spatial structures (‘spatially explicit protocols’) in reducing sampling requirements versus random sampling. Methods and Results: Canopy microclimate data were collected across multiple sites/seasons/training systems. Autocorrelation was found in all systems, with a periodicity generally corresponding to vine spacing. Three spatially explicit sampling models were developed to optimise the balance between minimum sample sizes and maximum fit to a known probability density function. A globally optimised explicit sampling (GOES) model, which performed multivariate optimisation to determine best‐case sampling locations for measuring fruit exposure, reduced fruit cluster sample size requirements versus random sampling by up to 60%. Two stratified sampling protocols were derived from GOES solutions. Spatially weighted template sampling (STS) reduced sampling requirements up to 24% when based on probabilistic panel weighting (PW), and up to 21% when preferentially selecting specific locations within canopy architecture (AW). Conclusions: GOES, PW STS and AW STS each reduced required sample size versus random sampling. Comparative analyses suggested that optimal sampling strategies should simultaneously account for spatial variability at multiple scales. Significance of the Study: This study demonstrates that dynamically optimised sampling can decrease sample sizes required by researchers and/or wineries.     

Composition of the glutenin macropolymer: effects of flour quality and nonamylolytic enzyme addition

The effects of the non-amylolytic enzymes pentosanase (PP), glucose oxidase (GLZ) and laccase (LAC), singly and in binary and ternary combinations, and wheat flour—a pure cultivar F0, and low- grade F1 and high-grade F2 commercial blends—on the electrophoretic pattern of the glutenin macropolymer (GMP) were investigated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE). GMP profiles of high (HMW), medium (MMW) and low (LMW) molecular weight glutenin subunits (GS) were correlated with dough and GMP characteristics and with bread quality and keepability.The total relative percentages of separated HMW, MMW and LMW-GS of GMP in control and enzyme-supplemented samples were: 33–37, 11–16 and 46–54% (F0), 47–74, 0 and 26–57% (F1) and 30–47, 15–18 and 32–38% (F2), respectively. HMW-GS/LMW-GS ratios, a quality index of glutenin, were found to range from 0.63 to 0.80 for all flour GMP. Enzyme addition induced moderate effects in pure wheat cultivar F0. Combinations of PP and GLZ (PPGLZ) allowed glutenin repolymerization after resting, leading to an increase of the bands of 104–107 and 88–90 kDa in GMP by SDS–PAGE. Analogously, PP plus LAC (PPLAC) and LAC alone increased the amount of the subunit of 81–82 kDa. Subunits of LMW increased with PPLAC and LAC, the subunit of 37–38 kDa having a higher percentage in GMP probably due to the presence of -, - and -gliadins besides the LMW-GS. Only the subunit of 81–82 kDa was positively associated with the dynamic rheological moduli G and G* and the subunit of 88–90 kDa correlated with a lower delta angle and, consequently, with a more elastic gel. After fermentation, the total percentage of subunits of HMW, MMW and LMW changed mainly in low-grade flour F1 (36–37, 17–19 and 14–19%) compared with high-grade flour F2 (32–37, 14–17 and 47–51%). The subunit of HMW with MW of 120–125 kDa (F1) and 93–95 kDa (F2) was positively correlated with the specific volume of breads and negatively with firmness of fresh and stored bread. Subunits of MMW negatively influenced the quality of fresh and stored bread of F1. The ratio HMW/LMW was positively correlated with bread firmness during staling in breads prepared with F2.     

Integrative and sealing-over recoveries from schizophrenia: distinguishing case studies

Fast-2, a membrane mutant of Paramecium aurelia, is due to a single-gene mutation and has behavioral abnormalities. Intracellular recordings through changes of external solutions were made. The mutant membrane hyperpolarized when it encountered solutions with low K+ concentration. This hyperpolarization and other associated activities were best observed in Ca- or Na-solutions devoid of K+. Membrane potential was plotted against the concentration of K+ (0.5 to 16 mM) in solutions of fixed Na+ or Ca++ concentration. The slopes of the curves for the mutant membrane were steeper than those for the wild type at the lower concentrations of K+. Inclusion of 2 mM tetraethylammonium chloride (TEA-Cl) counteracted the mutational effects. Spontaneous action potentials in Ba-solution and the electrically evoked action potentials in various solutions are normal in this mutant. We conclude that the resting permeability to K+ relative to the permeabilities to Na+ and Ca++ has been increased by the mutation.     

An Anglo-American comparison of the scaling of life events

Exact placement is an essential prerequisite for long-term use of a central venous catheter. Reported data show an extremely wide range of catheteral misplacements: from less than 1% to more than 60%. Some approaches appear to be less advantageous than others, but the highest rates of misplacement occur in the cubital, external jugular and saphenous veins. A series is presented of 378 radiographically controlled central venous catheters analysed for aberrant placement and loop formation. The total occurrence of faulty positioning and coiling reached 5.3%, while the respective incidences were 30% for the external jugular vein, 5.7% for the internal jugular vein, 5.5% for the infraclavicular technique of subclavian venepuncture, 5.3% for the innominate vein and 1.4% for the supraclavicular approach of subclavian venepuncture. The total frequency for pure loop formation was 2.9%. The authors discuss numerous reported data on catheter malpositioning, according to the specific techniques used, and compare them with thier own results. The relatively low incidence in the present series is possibly due to the high proportion of cases where the supraclavicular subclavian approach was used, the omission of the sphrenous/femoral and cubital techniques, and to pre-determining the length of the inserted catheteral segments.     

Response of the binding capacity of plasma testosterone-estradiol-binding globulin to norethindrone, 2 mg., and mestranol, 0.1 mg., in polycystic ovarian disease

Two cases of duodenal hematoma secondary to abdominal trauma in children are reported. The latent periods between trauma and the onset of symptoms were two and fifteen days. X-ray studies were conclusive for diagnosis. Pancreatic signs were mild in one case. Both cases were treated surgically, draining the fluid collection extramucosally. The authors advise surgical treatment whenever associated lesions need to be ruled out and when obstruction persists after the first days.     

Biodegradation of phthalic acid esters in river water and activated sludge

Infants born in New England with congenital heart defects were examined for association with time of year or population density. Moderate seasonal peaks in births were noted for complex ventricular septal defect, malposition defects, and transposition of the great arteries. Positive associations with population density were found for pulmonary atresia with ventricular septal defect, ventricular septal defect with secondary anomalies, and tricuspid atresia.     

Another hamster paradox: more males carry pups and fewer kill and cannibalize young than do females

The response of virgin male and female golden hamsters to young was studied. In contrast to most species, males are more likely to carry pups than are females. All males carried pups, but approximately 50% of females cannibalized the young. The females that did not cannibalize the pups carried them with less hesitation and after shorter latencies than did the males. The response of females to young was not correlated with the aggressiveness displayed toward adult males during separate tests. Tests with gonadectomized females indicated that the maintenance of pup-killing behavior is not dependent on concurrent gonadal hormones. Progesterone injections did not significantly increase pup killing in males that had previously carried young. Speculations on the adaptive significance of the male and female hamster's response to pups are presented.     

Examination of the ocular fundus with monochromatic light

In the process of insulin release by exocytosis, a fusion between the membranes of secretory vesicles and the plasma membrane occurs which is followed by the rupture of these membranes and the formation of an exocytotic stoma. As revealed by freeze-fracturing, the membrane structure is altered in the region of membrane interaction. An accumulation of membrane-associated particles can be observed in membranes of secretory vesicles contacting the plasma membrane. At the exocytotic stoma the boundary between granule limiting membrane and plasma membrane is occasionally outlined by a ring-like aggregation of membrane-associated particles. The effects of hypertonic solutions on tissue structure indicate that the membrane junctions during exocytosis are permeable to ions and small molecules, forming a communication between the interior of the secretory vesicle and the extracellular space.