Correlation of enzymatic,metabolic, and behavioral deficits in thiamin deficiency and its reversal

To clarify the enzymatic mechanisms of brain damage inthiamin deficiency, glucose oxidation, acetylcholine synthesis, and the activities of the three major thiamin pyrophosphate (TPP) dependent brain enzymes were compared in untreated controls, in symptomatic pyrithiamin-induced thiamin-deficient rats, and in animals in which the symptoms had been reversed by treatment with thiamin. Although brain slices from symptomatic animals produced¹⁴CO₂ and¹⁴C-acetylcholine from [U-¹⁴C]glucose at rates similar to controls under resting conditions, their K⁺-induced-increase declined by 50 and 75%, respectively. In brain homogenates from these same animals, the activities of two TPP-dependent enzymes transketolase (EC 2.2.1.1) and 2-oxoglutarate dehydrogenase complex (EC 1.2.4.2, EC 2.3.1.61, EC 1.6.4.3) decreased 60–65% and 36%, respectively. The activity of the third TPP-dependent enzyme, pyruvate dehydrogenase complex (EC 1.2.4.1, EC 2.3.1.12, EC 1.6.4.3.) did not change nor did the activity of its activator pyruvate dehydrogenase phosphate phosphatase (EC 3.1.3.43). Although treatment with thiamin for seven days reversed the neurological symptoms and restored glucose oxidation, acetylcholine synthesis and 2-oxoglutarate dehydrogenase activity to normal, transketolase activity remained 30–32% lower than controls. The activities of other TPP-independent enzymes (hexokinase, phosphofructokinase, and glutamate dehydrogenase) were normal in both deficient and reversed animals.Thus, changes in the neurological signs during pyrithiamin-induced thiamin deficiency and in recovery paralleled the reversible damage to a mitochondrial enzyme and impairment of glucose oxidation and acetylcholine synthesis. A more sustained deficit in the pentose pathway enzyme, transketolase, may relate to the anatomical abnormalities that accompany thiamin deficiency.Dedicated to Henry McIlwain.     

Field and laboratory observations of spawning periodicity and behavior of a northern population of the Atlantic silverside,Menidia menidia (Pisces: Atherinidae)

Synopsis The spawning periodicity and mating behavior of a northern population of the Atlantic silverside, Menidia menidia, was studied both in the field (Salem Harbor, Massachusetts) and in laboratory experiments. Spawning in the field coincided with new and full moons. Under conditions of unlimited food availability in artificial outdoor pools that received natural celestial illumination or in indoor aquaria receiving no evening illumination, spawning occurred every 1–3 days rather than fortnightly. These results suggest that tidal influences are a primary factor that synchronizes spawning in M. menidia. Published reports of diel time of spawning in both marine and freshwater species of Menidia indicate a generic tendency to spawn during mid-morning. This pattern coupled with high tides that cover suitable spawning substrates during mid-morning only at fortnightly intervals could account for the semilunar spawning cycle in marine populations of Menidia. Although general characteristics of the reproductive ecology of M. menidia in Massachusetts were similar to a thoroughly studied population in South Carolina, several differences were evident. The breeding season was shorter in Massachusetts (late; April–June), and occurred over a much lower range of temperatures (9–21°C). Intensity and frequency of spawning was correlated with the height of high tide. Eggs were deposited only on mats of intertidal, filamentous algae rather than on roots or stems of Spartina alterniflora or other intertidal vegetation. Promiscuous spawning occurred in small, highly male-dominated groups of fish, primarily after the tide had begun to recede, and often at the water's edge. No predation on spawning adults was observed but the mummichog, Fundulus heteroclitus, fed daily on developing embryos during high tide. Contribution No. 81, Massachusetts Cooperative Fisheries Research Unit, Contribution No. 349, Marine Sciences Research Center, State University of New York at Stony Brook.     

Biosynthesis of the yeast cell wall: Selective assays and regulation of some mannosyl transferase activities

Assays have been developed for some transfer reactions involved in the synthesis of Saccharomyces cerevisiae wall mannoproteins, both in a particulate preparation in the presence of EDTA or Triton X-100, and after lipid extraction with chloroform-methanol at -20C.The mannosyl transferase activities were also studied in cells made permeable to GDP-mannose by toluene-ethanol treatment (in situ). In these permeabilized cells, the glycosylating reactions dependent on lipid carriers (dolichol derivatives) did not function, but those independent of them were unaffected.The lipid-independent mannosyl transferase activities were partially inhibited by nucleotide diphosphates probably in a competitive manner. Increase of the nucleotide diphosphate pool in vivo might slow down the speed of the transfer reactions carried out by the mannan synthetase system.     

Nitrogen cycling in a15N-fertilized bean (Phaseolus vulgaris L.) crop

To increase our understanding of the fate of applied nitrogen inPhaseolus vulgaris crops grown under tropical conditions,¹⁵N-labelled urea was applied to bean crops and followed for three consecutive cropping periods. Each crop received 100 kg urea-N ha^?1 and 41 kg KCl?K ha^?1. At the end of each period we estimated each crop's recovery of the added nitrogen, the residual effects of nitrogen from the previous cropping period, the distribution of nitrogen in the soil profile, and leaching losses of nitrogen. In addition, to evaluate potential effects of added phosphorus on nitrogen cycling in this crop, beans were treated at planting with either 35 kg rock-phosphate-P, 35 kg superphosphate-P, or 0 kg P ha^?1. Results showed that 31.2% of the nitrogen in the first crop was derived from the applied urea, which represents a nitrogen utilization efficiency of 38.5%. 6.2% of the nitrogen in the second crop was derived from fertilizer applied to the first crop, and 1.4% of the nitrogen in the third crop. Nitrogen utilization efficiencies for these two crops, with respect to the nitrogen applied to the first crop, were 4.6 and 1.2%, respectively. In total, the three crops recovered 44.3% of the nitrogen applied to the first crop. The remainder of the nitrogen was either still in the soil profile or had been lost by leaching, volatilization or denitrification.¹⁵N enrichment of mineral-N(NO₃+NH₄) suggests that at the end of the second crop, the pulse of fertilizer applied to the first crop had probably passed the 120 cm depth.¹⁵N enrichment of organic-N suggests that root activity of beans and weeds transported nitrogen to 90–120 cm (or deeper). We could account for 109 kg fertilizer-N ha^?1 in harvested biomass, crop residue, and soil at the end of the first cropping period. This indicates an experimental error of about 10% if no nitrogen was lost by volatilization, denitrification, or leaching below 120 cm. At the end of the second and third crops, 76 and 80 kg N ha^?1, respectively, could be accounted for, suggesting that 20 to 25% of the applied-N was lost from the system over a 2-crop period. The two types of added phosphorus did not significantly differ in their effects on bean yields.     

Chemical enthalpy-entropy compensation effect in the hydrolysis of p-nitrophenyl carboxylates catalyzed by alkaline mesentericopeptidase

The temperature dependence of the hydrolysis of p-nitrophenyl carboxylates with general formula H(CH₂)_nCOOC₆H₄NO₂ catalyzed by alkaline mesentericopeptidase has been studied (n varying from 1 to 7, temperature range 2–30°C, pH 8.80, 5 vol% dimethylsulfoxide). The activation parameters of the deacylation step depend on the length of the hydrophobic side chain of the substrate molecule ( , , and decrease by 2.0 kcal/mol, 4.9 kcal/mol, and 10 eu, respectively, as the length of the acyl carbon chain increases from n = 1 to n = 4). The following criteria were applied to establish a chemical enthalpy-entropy compensation effect: (a) Exner's plot of log vs : (b) Petersen's plot of log, k/T vs 1/T; (c) Exner's statistical treatment in coordinates log k vs 1/T; (d) according to Krug et al. (ΔH^≠ vs ΔG_Thm^≠). By use of all the above-mentioned criteria the existence of a chemical enthalpy-entropy compensation effect was proved with an isokinetic temperature β of about 470°K, which is significantly higher than the average experimental temperature.     

Sympathetic control on salivary secretion by the parotid gland in rabbit. I. Effects on the unstimulated gland (author's transl)]

The sympathetic influences on the rabbit unstimulated parotid gland were studied. The experiments were carried out in anaesthetized rabbits with the Stenon aduct cannulated. Direct stimulation of the superior cervical ganglion elicits variable salivary flows. The high amylase content in the saliva points to a sympathetic secretory action upon acinar cells. The administration of alpha-adrenergic blocking agents (dihydroergotamine, phentolamine and phenoxybenzamine) clearly reduces and even abolishes the effect of the sympathetic stimulation upon flow. The administration of a beta-adrenergic blocking agent (propranolol) slightly reduces the sympathetic action. However the amylase activity is greatly reduced. All this suggests that the secretory effects on the fluid fraction should predominantly be alpha-adrenergic while on the secretion of enzymes the beta-receptors should play an important role.     

Studies on the adrenal cortex of hypophysectomized rats: A model for abnormal cellular atrophy and death

Summary Biochemical and ultrastructural studies indicate that the atrophy of adrenal cortex in hypoyhysectomized rats involves the following changes: (1) One to two days after hypophysectomy, there is loss of template activity resulting from cumulative DNA-damage and heterochromatinization.In vivo ACTH-administration led to recuperation of these cells, indicating damage during hypophysectomized state to be reversible. (2) If the duration of hypophysectomy is prolonged, some of the cells become irreversibly damaged and can no longer recuperate afterin vivo ACTH administration. (3) The period of most rapid cell death is from the third to seventh day after hypophysectomy. The cause of cell death is probably due to membrane damage in the absence of protein synthesis, leading to lysis of the cells. Lysozomes and macrophages are apparently not involved.Supported by U.S.P.H.S. grants AM-5384 and AM-13724 and taken in part from dissertations submitted by Chan and by Mostafapour to Wayne State University in partial fulfillment towards the Ph.D. degree.An invited article.     

Measurement of serum gentamycin levels by rapid tests

Gentamicin is a very useful antimicrobial agent for the treatment of serious infections caused by gram-negative bacteria. However, it's low therapeutic index and potential ototoxic and nephrotoxic side effects necessitate frequent determinations of serum concentration to assist in maintaining therapeutic levels and avoiding toxic levels. Two bioassays and a latex agglutination inhibition card (LAIC) test were evaluated to determine gentamicin levels in nearly 100 patient sera. Results were compared with a radioimmunoassay (RIA). Two bioassays, the Bio-Monitor and the GentaSak, gave correlation coefficients of 0.987 and 0.982, respectively. The correlation coefficient for the LAIC test was 0.987. All three tests compared well with RIA in accurately detecting gentamicin levels in patient as well as simulated sera. The LAIC test, however, was more rapid, giving results within half an hour whereas bioassays required 6–8 hours for completion. The LAIC test was also found to be more economical. It provides a suitable alternative to RIA procedures in small laboratories and for performing stat tests since batching is not necessary.