The transport of morphologically abnormal sperm in the female reproductive tract of mice

Mice of the PL/J strain exhibit a high percentage of morphologically abnormal sperm and provide a model for studying the function of abnormal sperm. The ability of such sperm to reach the site of fertilization within the female reproductive tract has been investigated. We have found a decrease in the percentage of structurally abnormal sperm within the population that reaches the oviduct. This observation suggests either that there is an active selection against abnormal sperm or that they are physiologically disadvantaged in reaching the site of fertilization.     

Leukotriene B4 binding to human neutrophils

[³H] Leukotriene B₄ (LTB₄) binds concentration dependency to intact human polymorophonuclear leukocytes (PMN's). The binding is saturable, reaches equilibrium in 10 min at 4°C, and is readily reversible. Mathematical modeling analysis reveals biphasic binding of [³H] LTB₄ indicating two discrete populations of binding sites. The high affinity binding sites have a dissociation constant of 0.46 × 10⁻⁹M and B_max of 1.96 × 10⁴ sites per neutrophil; the low affinity binding sites have a dissociation constant of 541 × 10⁻⁹M and a B_max of 45.6 × 10⁴ sites per neutrophil. Competitive binding experiments with structural analogues of LTB₄ demonstrate that the interaction between LTB₄ and the binding site is stereospecific, and correlates with the relative biological activity of the analogs. At 25°C[³H] LTB₄ is rapidly dissociated from the binding site and metabolized to 20-OH and 20-COOH-LTB₄. Purification of neutrophils in the presence of 5-lipoxygenase inhibitors significantly increases specific [³H] LTB₄ binding, suggesting that LTB₄ is biosynthesized during the purification procedure. These data suggest that stereospecific binding and metabolism of LTB₄ in neutrophils are tightly coupled processes.     

500 MHz H-NMR studies of bile salt-phosphatidylcholine mixed micelles and vesicles Evidence for differential motional restraint on bile salt and phosphatidylcholine resonances

¹H nuclear magnetic resonance (NMR) spectra at 500 MHz have been obtained for taurocholate/egg phosphatidylcholine mixtures of varying composition. The excellent chemical shift dispersion permits identification of most resonances for each component. This high-resolution character of the NMR spectra is retained until the phosphatidylcholine (PC) mole fraction exceeds 60–70% (the exact limit depends on ionic strength). ¹H linewidths have been monitored as a function of solute composition in order to evaluate trends in local molecular mobility of each component as the distribution of aggregate particles is varied, and to examine the effects of added NaCl in altering micellar size and shape. Although prior light scattering studies (Mazer, N.A., Benedek, G.B. and Carey, M.C. (1980) Biochemistry 19, 601–615) and our own work indicate a 6-fold increase in particle hydrodynamic radius from pure taurocholate micelles to 1 : 1 taurocholate/PC mixtures containing 150 mM NaCl, both lipid components retain substantial motional freedom and exhibit narrow NMR signals in this compositional region. As the solubilization limit for PC is approached (approx. 2:1 PC:taurocholate), differential behavior is observed for the two components: the motion of taurocholate becomes preferentially restricted, while polar portions of the PC remain mobile until large multilayers predominate.     

Characterization of a new marine methylotroph

Abstract A methanol-oxidizing bacterium from a marine environment has been isolated and characterized. The bacterium was a Gram-negative rod, capable of growth on methanol and methylamine, but not on multicarbon compounds. It showed a temperature optimum of 30°C, a salt optimum of 0.4% (w/v) and the mol % G + C of its DNA was 46%. Carbon was assimilated via the ribulose monophosphate pathway for formaldehyde fixation during growth on methanol. This bacterium superficially resembled other obligate methylotrophs requiring NaCl reported previously which were designated Methylomonas thalassica . It also appeared similar to many strains of obligate freshwater methylotrophs, except for its NaCl requirement and its lower mol % G + C.     

Cell culture of sixteen-day-old rat embryo cerebra and associated changes in ganglioside pattern

Abstract— Cortical slices from rat brain were incubated in Krebs-Ringer phosphate medium. Activity of the pyruvate dehydrogenase complex (PDH) was measured in homogenates of the incubated tissue. Increasing the extracellular KCI concentration from 5 to 75 mM caused a dose-dependent increase in activity of this rate-limiting mitochondrial enzyme. The increase in PDH activity, produced by high concentration of KCI. was associated with a decrease in the tissue content of ATP. Omission of calcium, or replacement of sodium by choline, reduced, and addition of ouabain prevented, the activation of the enzyme in the depolarized tissue.
The mechanism by which extracellular potassium can affect PDH activity is unknown. However, it is most likely that the alterations in enzyme activity are related to changes in properties of cell membranes during depolarization leading to intracellular events directly affecting the enzyme complex. These could include alterations in the concentrations of adenine nucleotides or free calcium ions in the cell.     

Isopropanol enhancement of carbon tetrachloride metabolism in vivo

We examined the effects of isopropanol (ISOP) pretreatment on the metabolism of ¹⁴CCl₄ to ¹⁴CO₂ and CHCl₃ exhaled in the breath, to ¹⁴C metabolite excreted in 24 hr urine and feces from 0 to 24 hr, and to ¹⁴C metabolite bound to liver at 24 hr. Fasted male rats were given 0.1 or 2.0 mmoles ¹⁴CCl₄/kg. ISOP pretreatment, which markedly enhanced the hepatotoxicity of CCl₄, selectively enhanced the rate and total extent of ¹⁴CO₂ and CHCl₃ metabolite exhalation. The pathways of CCl₄ metabolism leading to CO₂ and CHCl₃ metabolite formation may be more relevant to the hepatotoxicity of CCl₄ than the pathways leading to urinarym fecal or covalently bound metabolites.     

DIFFERENTIAL ENRICHMENT OF CELLS FROM EMBRYONIC RAT CEREBRA BY CENTRIFUGAL ELUTRIATION

—Centrifugal elutriation was used to obtain different populations of cells dissociated from 16-day-old rat embryo cerebra. The cell populations recovered were viable and could be maintained for several weeks in vitro. Sterile conditions were maintained throughout a preparation. Rat pups were removed by Caesarean section, the cerebra dissected and the cells dissociated by brief exposure to trypsin (0.125%, 6 min). An equivalent volume of elutriation medium (Dulbecco's medium containing 1% fetal calf serum, sodium bicarbonate, penicillin and streptomycin, EDTA, and deoxyribonuclease) was added to the trypsin-cell suspension, the dissociated cells pelleted, resuspended in elutriation medium and counted. Up to 4 x 10⁸ cells were injected into the previously sterilized elutriator. Seven fractions were usually recovered from a preparation. The first fraction contained primarily red blood cells and cell debris, which could not be maintained in vitro. Upon culture, fraction 2 consisted of predominantly non-neuronal cells, while fractions 3–6 contained neuronal and non-neuronal cells. The morphological characteristics of the neurons differed in these fractions. Fraction 7 contained cells that had reaggregated during the elutriation procedure and exhibited a variety of cell types in vitro.