The fine structure of the terminal segment of the bovine seminiferous tubule

Summary The intratesticular excurrent duct system of the bull is composed of rete testis, tubuli recti, and the terminal segment of the seminiferous tubules. Each terminal segment is surrounded by a vascular plexus and may be subdivided into a transitional region, middle portion, and terminal plug. The modified supporting cells of the middle portion and the terminal plug no longer display the typical Sertoli-Sertoli junctions seen in the transitional region and the seminiferous tubule proper. In the region of the terminal plug a distinct central lumen is generally not observed: spermatozoa and tubular fluid must pass through an intricate system of communicating clefts between the apices of the closely attached modified supporting cells. Vacuoles in the supranuclear region of the cells in the middle portion indicate strong transepithelial fluid transport. In analogy to the epithelium of rete testis and tubuli recti, the supporting cells of the terminal segment are capable of phagocytosing spermatozoa. The vascular plexus investing the terminal segment serves a dual purpose: it is a regulatory device for fluid and sperm transport, as well as an area of increased diapedesis for white blood cells.Supported by a grant from the Deutsche Forschungsgemeinschaft     

External calcium concentration and fertilization in the sea urchin, Strongylocentrotusfranciscansus (A. Agassiz) (Echinodermata)

The role of external calcium ions in the fertilization of the sea urchin, Strongylocentrotusfranciscansus (A. Agassiz), was studied. When eggs were inseminated with jelly-treated sperm in artificial sea waters containing various concentrations of calcium, the percentage of fertilization decreased with decreasing concentrations of calcium. A small amount of external calcium ions was also found to be essential for fertilization by spermatozoa with reacted acrosomes. The binding capacity of jelly-treated spermatozoa to eggs was, however, not adversely affected by a calcium deficiency. These results suggest that calcium ions are required not only for the initiation of the acrosome reaction, but also for successful levels of fertilization following sperm-binding to eggs even after the acrosome reaction has occurred.     

Transposition of the gene cluster CYC1-OSM1-RAD7 in yeast

A mutant of the yeast Saccharomyces cerevisiae contains an increased amount of iso-1-cytochrome c because two copies of a segment, denoted COR, were transposed to a new position on chromosome VII, while the original COR region was retained at the normal position on chromosome X; this COR segment encompasses the CYC1, OSM1 and RAD7 loci which determine, respectively, iso-1-cytochrome c, osmotic sensitivity and ultraviolet light sensitivity. The analysis of genomic DNA with cloned probes indicates that the length of the COR segment is approximately 12,000 base-pairs. We suggest that certain normal strains of yeast, which possibly may contain reiterated sequences, can produce extended transpositions similar to prokaryotes.     

Heparin enhances fibrinolysis in B16 mouse melanoma cells

The fibrinolytic activity of two tumorigenic B16 mouse melanoma lines was stimulated by exogenous hog mucosal or beef lung heparin. In contrast, the activity of two normal fibroblast lines was unaffected. The degradation of ¹²⁵l-fibrin was increased up to 3.6-fold by the addition of heparin. Chondroitin-4-sulfate or dextran sulfate did not change the fibrinolytic activity of three of the cell lines, but, at concentrations where enhancement by heparin was much reduced, the activity of one of the B16 melanoma lines was somewhat elevated. Antithrombin III did not alter the plasminogen activator activity of the B16 cell lines, but, in the presence of exogenous heparin, the enhancement of fibrinolysis was greatly reduced. The polymers were not cytotoxic during the assay period, and, had little affect on the plating efficiencies of the lines.     

Nickel requirement for chemolithotrophic growth in hydrogen-oxidizing bacteria

In a comparative study the requirement of several strains of autotrophic hydrogen-oxidizing bacteria for nickel was examined. Autotrophic growth was studied both in liquid media, previously freed from trace metals; and on solidified media, using a plate diffusion assay. The latter assay was based on the observation that EDTA causes complete inhibition of autotrophic growth on agar medium as a result of nickel deficiency. Nickel was shown to be required as a trace element in five strains of Alcaligenes eutrophus, in two strains of Xanthobacter autotrophicus, in Pseudomonas flava, in Arthrobacter spec. 11X and in strain 12X. In these bacteria nickel was not replaceable by cobalt, copper, manganese or zinc ions. No significant nickel requirement was detected by these methods, however, for Paracoccus denitrificans and Nocardia opaca 1b.     

Enhancement of cellulose accessibility and enzymatic hydrolysis by simultaneous wet milling

It has been shown that the rate of enzymatic saccharification of cellulosic materials including “pure” cellulose (Whatman CF?11 cellulose), newsprint, lignocellulose (prehydrolyzed to remove hemicelluloses), and wood can be substantially increased by simultaneous wet milling. An enhanced hydrolysis rate was sustained above that observed for ball milling: providing a more extensive saccharification. The cellulosic substrates were wet milled with a variety of grinding elements, such as sand, glass beads, and stainless-steel beads, agitated in a shaker bath. Simultaneous hydrolysis was achieved with a 2% substrate slurry in a 0.1M acetate buffer at 45°C and pH 5. The effectiveness of this process was dependent upon the lignified matrix of the cellulose microfibrils, the grinding elements, and the oscillation frequency of the shaker bath. Wet milling “pure” cellulose for 48 hr, with 3.5 mm glass beads and 200 oscillations/min (opm), yielded 1031 mg reducing sugar/g substrates (93% saccharification) as compared to 483 mg (44%) for the ball-milled sample and 253 mg (23%) for the unmilled material. With the lignified substrates stainless-steel beads (3.5 mm) were more effective than glass. For lignocellulose 529 mg sugar/g substrate (93% saccharification) could be obtained by wet milling with cellulase for 24 hr. This was about three times greater than that of the ball milled (169 mg, 30%) and 10 times greater than that of the unmilled (52 mg, 9%) substrates. The method was also effective for wood particles (60 mesh) giving 143 mg sugar/g wood (approximately 38% saccharification) in 48 hr, whereas the ball-milled sample gave only 79 mg (21%) and the unmlilled substrate 38 mg (10%). These observations can be explained on the basis of the current crystalline theory for the morphology of the cellulosic microfibrils. The advantage of wet milling and simultaneous hydrolysis apparently depends on a continuous generation of accessible sites and sustained rapid hydrolysis rate as the saccharification proceeds, where in the pretreated substrates the hydrolysis rate slow down as the active sites are reduced.     

A technique for quantitating enzyme histochemistry in adjuvant arthritic joints. II. Acid phosphatase

Synopsis This paper reports on the incorporation of acid phosphatase histochemistry with a quantitative technique designed to measure the percentage of histochemically-localized enzyme-reactive cells found in adjuvant arthritic articular cartilage, synovial membrane and bone marrow. With the data presented and the comparisons made, this incorporation is validated and adjuvant arthritic acid phosphatase lesions established. In addition to discussing the importance of the histochemical data and its relationship with intra-articular lysosomes, the applicability of the reactive cell values to drug inhibition studies is discussed.     

The influence of coal ash and thermal discharges upon the distribution and bioaccumulation of aquatic invertebrates

The distribution, density and uptake of twenty elements by aquatic invertebrates inhabiting a drainage system, that received excessive coal ash effluent (275 JTU of turbidity) at one end and thermal loading (44.5°C) at the other end, was studied for 15 months. The ash settling basin filled during the first eight months of sampling which resulted in the release of ash effluent directly into the receiving system. Density of invertebrates was lowest in the 300 m stream between the ash basin and swamp and highest 1200 m beyond the stream-swamp confluence where ash influence was minimal. Invertebrate density was lowest in the stations where turbidity from ash effluent was greatest. The most tolerant invertebrates to coal ash stress were odonates (Libellula sp. and Enallagma sp.), crayfish (Procambarus sp.), amphipods (Gammarus sp.) and gastropods (Physa. sp.), and midges (Chironomidae) when the basin was filling. During the period of ash overflow, all groups were either reduced in numbers or absent. In the thermally stressed station, Libellula sp. was the predominant invertebrate sampled when water temperature ranged from 25.5–45.5°C (257-1=28.7°C) all aquatic invertebrates were limited in numbers and density when temperature exceeded the lower and upper ranges of 10.0–38.0°C.This research was supported by AEC Contract AT (38-1-824)This research was supported by AEC Contract AT (38-1-824)