A comparison of sperm morphology and silver nitrate staining characteristics in the domestic ferret and the black-footed ferret

Ejaculated sperm from the domestic ferret (Mustela putorius furo) and the black-footed ferret (Mustela nigripes) were compared for differences in morphological abnormalities and argentophilic protein distribution. Thawed domestic ferret sperm was also compared to fresh sperm to determine whether there were any effects on cell morphology due to cryopreservation. There were statistically significant differences between the two species of ferret in two of the categories scored. The domestic ferret had a higher frequency of cells that were bent in the midpiece and in the principal piece, and a higher frequency of headless and tailless cells when compared to the black-footed ferret. There were no statistically significant differences in cell morphology between the fresh and cryopreserved ejaculates of the domestic ferret employing a standard egg yolk cryoextender. Silver nitrate staining distribution was different between the two species in both the head and tail region.     

The Site Specificity of Two Species of Gregarina in Tenebrio molitor Larvae

The site specificity of the apicomplexans Gregarina cuneata and Gregarina sleini , in larval Tenebrio molitor was investigated. Gregarina cuneata was found to inhabit the anteriormost region of the larval midgut, while G. steini was restricted to the posterior portion of the intestine. The site specificity of the pair was conserved in single and concurrent infections. Interspecific interactions do not seem to be presently responsible for the resource partitioning by the 2 gregarine species. Key words. Gregarina, site specificity, Tenebrio molitor.     

Intracranial self-stimulation motivates treadmill running in rats.

Most animal running models have traditionally used aversive motivators to induce exercise tasks. This study demonstrates treadmill running motivated by reinforcement of intracranial self-stimulation (ICSS), providing an alternative model with which to study physiological responses to exercise. Twenty-nine male Sprague-Dawley rats were stereotaxically implanted with bipolar electrodes aimed at the ventral tegmental area of the brain. After 7 days of operant lever-press training for ICSS, rats that pressed at least 50 presses/min were randomly divided into three conditions: exercise-reinforcing brain stimulation (Ex-St), exercise-aversive shock (Ex-Sh), and sedentary controls (C). Ex-St and Ex-Sh ran for 30 min at 25 m/min at 5% grade for 2 wk with ICSS and electric shock as the motivator, respectively, while C did not run. At the end of 2 wk, Ex-St and Ex-Sh performed an endurance run. Results show that Ex-St ran longer than Ex-Sh [63 +/- 10 vs. 42 +/- 10 (SD) min; P less than 0.05]. HR was higher in Ex-St than in C (P less than 0.05). Rectal temperature increased similarly in both exercise groups. This model provides a highly effective method to motivate treadmill running in rats and as such can be used to characterize physiological responses to exercise without the potentially confounding influence of stress associated with an aversive shock motivator.     

Cyclic Variations in Nitrogen Uptake Rate in Soybean Plants: Uptake During Reproductive Growth

Net uptake of by non-nodulated soybean plants [Glycme max(L ) Merr cv Ransom] growing in flowing hydroponicculture was measured daily during a 63 d period of reproductivedevelopment between the first florally inductive photopenodand late seed growth Removal of from a replenished solution containing 10 mol m^– was determined by ion chromatography Uptake of continued throughout reproductive development The net uptakerate of cycled between maxima and minima with a periodicity of oscillation of 3 to 7 d during the floralstage and about 6 d during the fruiting stage. Coupled withincreasing concentrations of carbon and C:N ratios in tissues,the oscillations in net uptake rates of are evidence that the demand for carbohydrate by reproductiveorgans is contingent on the availability of nitrogen in theshoot pool rather than that the demand for nitrogen followsthe flux of carbohydrate into reproductive tissues. Key words: Nitrate uptake rate, carbon-nitrogen partitioning, Glycme max (L ) Merrill     

Carbon Dioxide Effects on Carbohydrate Status and Partitioning in Rice

The atmospheric carbon dioxide (CO₂) concentration has beenrising and is predicted to reach double the present concentrationsometime during the next century. The objective of this investigationwas to determine the long-term effects of different CO₂ concentrationson carbohydrate status and partitioning in rice (Oryza sativaL cv. IR-30). Rice plants were grown season-long in outdoor,naturally sunlit, environmentally controlled growth chamberswith CO₂ concentrations of 160, 250, 330, 500, 660, and 900µmolCO₂ mol¹ air. In leaf blades, the priority between the partitioningof carbon into storage carbohydrates or into export changedwith developmental stage and CO₂ concentration. During vegetativegrowth, leaf sucrose and starch concentrations increased withincreasing CO₂ concentration but tended to level off above 500µmolmol^–1 CO₂. Similarly, photosynthesis also increased withCO2 concentrations up to 500µmol mol^–1 and thenreached a plateau at higher concentrations. The ratio of starchto sucrose concentration was positively correlated with theCO₂ concentration. At maturity, increasing CO₂ concentrationresulted in an increase in total non-structural carbohydrate(TNC) concentration in leaf blades, leaf sheaths and culms.Carbohydrates that were stored in vegetative plant parts beforeheading made a smaller contribution to grain dry weight at CO₂concentrations below 330µmol mol^–1 than for treatmentsat concentrations above ambient Increasing CO₂ concentrationhad no effect on the carbohydrate concentration in the grainat maturity Key words: CO₂ enrichment, starch, sucrose     

A Dynamic Growth Model of Vegetative Soya Bean Plants: Model Structure and Behaviour under Varying Root Temperature and Nitrogen Concentration

A differential equation model of vegetative growth of the soyabean plant (Glycine max (L.) Merrill cv. ‘Ransom’)was developed to account for plant growth in a phytotron systemunder variation of root temperature and nitrogen concentrationin nutrient solution. The model was tested by comparing modeloutputs with data from four different experiments. Model predictionsagreed fairly well with measured plant performance over a widerange of root temperatures and over a range of nitrogen concentrationsin nutrient solution between 0.5 and 10.0 mmol in the phytotron environment. Sensitivity analyses revealedthat the model was most sensitive to changes in parameters relatingto carbohydrate concentration in the plant and nitrogen uptakerate. Key words: Glycine max (L.) Merrill, dry matter, nitrogen uptake, partitioning, photosynthesis, respiration, sensitivity analysis     

Ribulose 1, 5-bisphosphate Carboxylase/ Oxygenase Activities in Leaves of Greenhouse Roses

An enzyme assay was developed to measure the initial and Mg²⁺–CO₂activated forms of Ribulose 1,5-bisphosphate Carboxylase/Oxygenase(Rubisco) in rose leaves. The assay was verified by co-extractionof the leaflets with partially purified spinach Rubisco andthrough correlation with net photosynthetic rates of individualleaflets (r²=0.7324). Changes in activities were measured asa function of depth of leaves in the canopy for two cultivarsof greenhouse hybrid tea roses. Initial Rubisco activity declinedwith increasing canopy depth for both cultivars. The activatedform of the enzyme, however, remained constant with canopy depthfor cv. Red Success; but increased with canopy depth, then declinedafter mid-canopy in the cv. Royalty. Rubisco activities werealso measured in the cv. Red Success grown in CO₂ enriched environments(100 mm³ dm^–3) at three humidity levels. The activitieswere not significantly affected by humidity treatment. However,there was a trend for plants grown at lower humidity to havehigher activated activities. Key words: Humidity, Rubisco, Rosa ? hybrida, Royalty, Red Success     

Cyanide and methylisocyanide binding to the isolated iron-molybdenum cofactor of nitrogenase

19F NMR and x-ray absorption experiments have been performed with both the isolated FeMo cofactor and the MoFe protein of nitrogenase in search of direct evidence for substrate or inhibitor binding. Using 19F NMR as a probe and p-CF3C6H4S- as the receptor ligand, the data show that the nitrogenase inhibitors CN- and CH3NC bind to the isolated FeMo cofactor-RFS- complex in N-methylformamide with a finite formation constant. Their binding increases the electronic relaxation time of the complex and increases the life-time of the FeMo cofactor-p-CF3C6H4S- bond, Parallel molybdenum K edge and extended x-ray absorption fine structure experiments show that CH3NC does not bind to molybdenum. Although CO and N3- both relieve CN- and CH3NC inhibition of electron flow through nitrogenase, unlike the latter, they do not appear to bind to isolated FeMo cofactor. In experiments with the dithionite-reduced MoFe protein, we did not detect any changes in the molybdenum K edge or extended x-ray absorption fine structure spectra upon addition of CO, N2, C2H2, NaCN, CH3NC, or azide demonstrating that either these substrates and inhibitors do not bind to molybdenum or that the FeMo cofactor site of nitrogenase is inaccessible to substrate binding except under turnover conditions.     

Free sphingosine formation from endogenous substrates by a liver plasma membrane system with a divalent cation dependence and a neutral pH optimum

Long-chain (sphingoid) bases may serve as another category of "lipid second messenger" because they inhibit protein kinase C and affect multiple cellular functions. Free sphingosine has been found in rat liver (Merrill, A. H., Jr., Wang, E., Mullins, R. E., Jamison, W. C. L., Nimkar, S., and Liotta, D. C. (1988) Anal. Biochem. 171, 373-381); hence, this study determined if liver plasma membranes contain free long-chain bases and have the ability to form them from endogenous enzymes and substrates. Isolated plasma membranes contained 0.45 nmol of sphingosine/mg of protein which, based on the recovery of the membranes, was equivalent to 3.5 +/- 1.2 nmol/g of liver and at least half of the total free sphingosine in liver. When the membranes were incubated at 37 degrees C, the amount increased at an initial rate of 5-25 pmol/min/mg, resulting in a 2-3-fold increase over an hour. Sphingosine formation required divalent cations, was optimal at neutral to alkaline pH, and was temperature-dependent. Activities with these characteristics were not identified in microsomes or lysosomes (lysosomal activities with acidic pH optima were detected, however); hence, they appear to reflect a separate plasma membrane system. Sphingosine formation was stimulated by ceramides either added exogenously or formed endogenously by treating the membranes with sphingomyelinase (but not endoglycoceramidase). Sphingomyelin hydrolysis to ceramide was also observed during incubation of the plasma membranes alone. Some of the properties of this system resembled the neutral sphingomyelinase and ceramidase activities of liver. While the physiological significance of this endogenous sphingosine is not known, this system has the appropriate subcellular location to provide sphingosine as a participant in signal transduction.     

A putative ATP binding protein influences the fidelity of branchpoint recognition in yeast splicing

We previously described a dominant suppressor of the splicing defect conferred by an A----C intron branchpoint mutation in S. cerevisiae. Suppression occurs by increasing the frequency with which the mutant branchpoint is utilized. We have now cloned the genomic region encoding the prp16-1 suppressor function and have demonstrated that PRP16 is essential for viability. A 1071 amino acid open reading frame contains sequence motifs characteristic of an NTP binding fold and further similarities to a superfamily of proteins that includes members with demonstrated RNA-dependent ATPase activity. A single nucleotide change necessary to confer the prp16-1 suppressor phenotype results in a Tyr----Asp substitution near the "A site" consensus for NTP binding proteins. We propose that PRP16 is an excellent candidate for mediating one of the many ATP-requiring steps of spliceosome assembly and that accuracy of branchpoint recognition may be coupled to ATP binding and/or hydrolysis.