Two isozymes of dihydroxyacetone phosphate reductase in dunaliella

Two isoforms of dihydroxyacetone phosphate reductase were present in Dunaliella tertiolecta. The major form was located in the chloroplast and the minor form in the cytosol. The chloroplastic reductase eluted first from a DEAE cellulose column followed immediately by the cytosolic form. Both forms were unstable and cold labile. Addition of 5 millimolar dithiothreitol helped to stabilize the enzymes. The cytosolic isoform of DHAP reductase was detected only if the cells were in an active log phase of growth. Then its activity was 20 to 30% of the total reductase activity. When cell cultures entered late log phase of growth the activity of the cytosolic form of the enzyme disappeared, but the chloroplastic form remained. The cytosolic DHAP reductase from Dunaliella has some properties similar to the cytosolic isoform from spinach leaves. Detergents inhibited both enzymes. However, neither form of the algal dihydroxyacetone phosphate reductase was stimulated by fructose 2,6-bisphosphate. In Dunaliella the properties of the chloroplastic form were those expected for glycerol production for osmoregulation, whereas the cytosolic form, like the reductases in leaves, is more likely involved in glycerol phosphate formation for lipid synthesis.     

Cytochemical localization of calcium in renal sac nephrocytes of Helix aspersa

In renal sac nephrocytes of Helix aspersa, intracellular calcium has been localized using the oxalate-pyroantimonate (OPA) and phosphate-pyroantimonate (PPA) methods. Pyroantimonate precipitates are preferentially localized in the excretory spherule and in vesicles located in the basal and lateral regions of the nephrocyte. Such vesicles appear to release their content into the excretory vacuole. Calcium may interact with diverse types of molecules present in the excretory vacuole, thus favouring stabilization and packaging of the excretory spherule.     

The effects of estrous cow serum on the in vitro maturation and fertilization of the bovine follicular oocyte

The effect of serum obtained from a cow at the time of standing estrus (serum A), at ovulation (serum B), and at 24 h after ovulation (serum C) on the in vitro maturation and fertilization of bovine oocytes was examined. Of 144 (Group A), 159 (Group B), and 158 (Group C) oocytes, 77 (53.4%), 82 (51.6%) and 82 (51.9%) oocytes were characterized by expansion of cumulus cells, respectively. There was no significant difference in the effect of the three types of cow serum on the cumulus expansion (P < 0.05). Of 461 oocytes, 316 oocytes were cultured with sperm for fertilization, and 145 oocytes were cultured without sperm for evidence of parthenogenetic development. Of 56 (Group A), 56 (Group B), and 62 (Group C) oocytes with expanded cumulus cells, 19 (33.9%), 7 (12.5%), and 11 (17.7%) oocytes were cleaved, respectively, after exposed to the sperm for 24 h. There was a significant difference in the effect of the three types of cow serum on the fertilization rate (P < 0.05). A total of 145 oocytes was cultured in the absence of sperm and no evidence of parthenogenetic division was observed. The effect of the three types of serum obtained from the cow on the maturation of oocytes was not significant, but a significant difference did exist in the fertilization rate of oocytes. Cow serum obtained at the time of standing estrus had a beneficial effect on the fertilization rate of oocytes in vitro.     

Changes in quality of stallion spermatozoa during cryopreservation: Plasma membrane integrity and motion characteristics

Better procedures for freezing and thawing equine sperm are needed since variable fertility is obtained when cryopreserved sperm are used. To evaluate current methods of freezing equine sperm, we examined spermatozoal quality by means of two new techniques. These measured the integrity of plasma-acrosomal membranes by immunofluorescent analyses of binding of an antibody specific to the acrosome and evaluated eight parameters of spermatozoal motion using a fully automated computerized system. Five ejaculates from each of eight stallions were processed for freezing in egg yolk-lactose extender with 4% glycerol. Spermatozoal quality was assessed at four different points: at less than 15 min after collecting and before processing (Step 1); after centrifugation and just before freezing (Step 2); immediately after thawing less than 3 h after freezing (Step 3); and immediately after thawing 10 to 20 d after freezing (Step 4). Acrosome-specific monoclonal antibody detected differences (P <0.05) among steps and ejaculates within stallions. All parameters of spermatozoal motion, including the percentage of motile sperm, percentage of progressively motile sperm, curvilinear velocity, straight line velocity, linearity, amplitude of lateral head displacement, and radius of the average path for circularly swimming sperm, differed (P <0.05) among steps, and most of these parameters differed among ejaculates within a stallion and among stallions. For Steps 2 and 3, 62 and 37% of the sperm were motile, and 56 and 23% of all motile sperm had a curvilinear velocity of >100 mum/sec. Most damage to sperm occurred as a result of freezing-thawing, whereas centrifugation of sperm caused only minor damage.     

Comparisons of eight commercial on-farm milk progesterone tests

To evaluate eight commercial on-farm milk progesterone kits, milk samples (50 ml each of foremilk and postmilk strippings) were collected during the estrous cycle from 10 cycling Holstein cows for 24 consecutive days. Relative concentrations of progesterone were classified as low or high by comparison with standard progesterone samples supplied with each kit. The concentration of progesterone in each milk sample was determined by radioimmunoassay (RIA). Accuracy of classification into low or high levels by commercial tests was determined by the percentage of similarity with RIA values using discriminant analysis. Accuracy of the eight tests ranged from 89.0 to 98.9% for low progesterone, 74.8 to 85.6% for high progesterone, and 80.3 to 87.3% for all samples (n = 238). The percentage of fat in milk or an interaction of the percentage of milkfat by day of estrous cycle influenced commercial test results for all tests except Accufirm and Calfcheck. Progesterone levels, estimated by the test-kits, were low from 1.5 +/- 0.5 to 2.8 +/- 0.9 days before estrus (X +/- SEM) and until 4.0 +/- 0.6 to 5.9 +/- 1.3 days after estrus. These data support the principle that a single low progesterone sample cannot be used to determine proper timing of insemination. All eight commercial kits can be used to determine accurately the relative concentrations of progesterone in milk samples.     

Ultrastructure of the domestic tom cat (Felis domestica ) and tiger (Panthera tigris altaica ) spermatozoa

The ultrastructure of spermatozoa from the domestic tom cat and the Siberian tiger was studied. Semen was collected from anesthetized tom cats and Siberian tigers by electroejaculation. Spermatozoa were fixed and processed for examination by transmission electron microscopy. The principle differences between the spermatozoa from the two species were the head shape, mitochondrial organization in the neck area and structure of the fibrous sheath. Tom cat spermatozoa had an elongated oval-shaped head, while tiger spermatozoa had a more rounded head shape. Circularly oriented mitochondria in the neck area, near the proximal centriole, were frequently observed in tiger cells but rarely observed in tom cat cells. The semicircular ribs of tom cat spermatozoa were larger than the ribs of tiger spermatozoa. Also, the dense fibers (Numbers 3 and 8) of the corresponding microtubule doublets were fused or connected to the longitudinal columns in tiger spermatozoa but showed only occasional attachment in tom cat spermatozoa. These differences could influence results when the tom cat is used as a model for studying tiger semen.     

Infectious disease and the Unesco basic education initiative

As part of a Basic Education for All campaign, Unesco is exploring ways to improve health and nutrition in school-age children in order to help them take advantage of the only schooling they may ever receive. At the first international Unesco working meeting on health, nutrition and school performance, intestinal helminths were identified as more appropriate immediate targets than malaria and other infectious diseases. Prevalence and intensity of infection in school-age children, the schools as vehicles of intervention, and the availability of interventions contributed to this choice. However, there are still many gaps in our knowledge of the effects of infectious diseases in schoolchildren. Through the improvement of understanding in this area, the parasitology community may be able to contribute to the Unesco project.     

Supply-side ecology and benthic marine assemblages

Many marine invertebrates have a planktonic stage of their life history during which widespread dispersal and much mortality occur. The numbers surviving to recruit into habitats occupied by adults are therefore very variable in time and space. Models for the structure and dynamics of benthic assemblages tend to focus on processes causing death - often assuming consistent arrivals of recruits. Supply-side ecology is a newly fashionable term to describe recent interest in the long-realized consequences of variations in recruitment. Such variations have important influences on theory and empirical research in these assemblages.     

Reactive extraction of penicillin G from mycel-containing broth in a countercurrent extraction decanter

Penicillin was recovered from mycel-containing fermentation broth by direct reactive extraction into a counter-current extraction decanter, Type CA 226-290 of the Westfalia Separator Co., at room temperature via steady state operation. Penicillin concentrations in the feed varied from 3 to 41 g L(-1), Amberlite LA-2 carrier concentrations from 7 to 20 g L(-1)and/or DITDA carrier concentrations from 7.2 to 84 g L(-1), the LA-2-to-penicillin mole concentration ratio from 4 to 6.4, and/or the DITDA-to-penicillin mole concentration ratio was maintained at 2. The throughputs of the fermentation broth (520 to 880 L h(-1)) of the solvent phase (200 to 860 L h(-1)) and the over all throughput (800 to 1750 L h(-1)) were high. Extraction degrees of 72 to 96% were achieved between pH 4.6 and 5.1. Without carriers in the same pH range, extraction degrees of only 17 to 19% were attained. By reducing the pH to 2.3 and in the absence of carriers, the degree of extraction was increased to 61%. However, during the extraction, 6.5% of the penicillin decomposed. At these high throughputs, the steady state was attained within 1 to 4 min. Through the mechanical stress, the length of the hyphae was reduced and the protein content of the broth was increased by 50 to 100%. However, this protein content had no appreciable influence on the phase separation.