Guanosine triphosphate catabolism in purine nucleoside phosphorylase deficient human B lymphoblastoid cells

GTP catabolism induced by sodium azide or deoxyglucose was studied in purine nucleoside phosphorylase (PNP) deficient human B lymphoblastoid cells. In PNP deficient cells, as in control cells, guanylate was both dephosphorylated and deaminated but dephosphorylation was the major pathway. Only nucleosides were excreted during GTP catabolism by PNP deficient cells and the main product was guanosine. The level of nucleoside excretion was largely affected by intracellular orthophosphate (P_i) level. In contrast, normal cells excreted nucleosides only at low P_i level while at high P_i levels, purine bases (guanine and hypoxanthine) were exclusively excreted. PNP deficiency had no effect on the extent of GMP deamination.     

Effects of nitrogen deficiency and light of high intensity onCryptomonas rufescens (Cryptophyceae)

Summary C.rufescens excystment, experimentally induced, corresponds to a general metabolism recovery of the cell, previously in a resting phase. The cytoplasm changes without any polarity, and organelles like gullet and flagella redifferentiate. The thylakoids develop mainly from the stored lipidic compounds which then disappear. Phycoerythrin immediately fills the intrathylakoidal lumen. Pigment synthesis seems closely associated with the development of membranes. The activated cell divides and the cyst wall breaks down. The destruction of the wall begins in the median layer and is followed by a mechanical rupture of the external and internal layers. Each germinative cyst releases two or four fully differentiated cells. There is an exact symmetry between excystment and encystment, all the transformations of theC. rufescens cell being reversible.     

Influence of plant nutrient concentration on growth rate: Use of a nutrient interruption technique to determine critical concentrations of N,P and K in young plants

A method is described for determining the way in which growth rate varies with plant nutrient concentration using a simple nutrient interruption technique incorporating only 2 treatments. The method involves measuring the changes in growth and nutrient composition of otherwise well-nourished plants after the supply of one particular nutrient has been withheld. Critical concentrations are estimated from the relationship between the growth rate (expressed as a fraction of that for control plants of the same size which remained well-nourished throughout) and the concentration of the growth-limiting nutrient in the plants as deficiency developed. Trials of the method using young lettuce plants showed that shoot growth rate was directly proportional to total N (nitrate plus organic N) concentration, and linearly or near-linearly related to K and P concentration over a wide range; the corresponding relationship for nitrate was strongly curvi-linear. Critical concentrations (corresponding to a 10% reduction in growth rate) determined from these results were similar to critical values calculated from models derived from field data, but were generally higher than published estimates of critical concentration (based on reductions in shoot weight) for plants of a similar size. Reasons for these discrepancies are discussed. Nitrate, phosphate or potassium concentrations in sap from individual leaf petioles were highly sensitive to changes in shoot growth rate as deficiency developed, with the slope of the relationships varying with leaf position, due to differences both in their initial concentration and in the rates at which they were utilized in individual leaves. Each nutrient was always depleted more quickly in younger leaves than in older ones, providing earlier evidence of deficiency for diagnostic purposes. Although the plants were capable of accumulating nitrate, phosphate and potassium well in excess of that needed for optimum dry matter production during periods of adequate supply, the rate of mobilization of these reserves was insufficient to prevent reductions in growth rate as the plants became deficient. This brings into question the validity of the conventional concept that luxury consumption provides a store of nutrients which are freely available for use in times of shortage. The implications of these results for the use of plant analysis for assessing plant nutrient status are discussed.     

Developmental Changes in Glycolipid Synthesizing Enzymes in the Brain of a Myelin-Deficient Mutant Wistar Rat

Changes in the activities of UDP-galactose:ceramide galactosyltransferase (CGalT, EC 2.4.1.45), UDP-glucose:ceramide glucosyltransferase (CGlcT, EC 2.4.1.80) and 3'-phosphoadenosine-5'-phosphosulfate (PAPS): galactosylceramide 3'-sulfotransferase (EC 2.8.2.11) over the myelinating period between 12 and 25 days were studied in the brains of control and myelin-deficient rats. Although the activity of galactosyltransferase with ceramides containing hydroxy fatty acids quadrupled in normal male littermates between 14 and 20 days, hardly any increase was observed in the mutant and the activity was less than 10% of control above 20 days of age. With normal fatty acid containing ceramides as acceptors, the activity decreased from 83% of the control at 12 days to approximately 30% after 20 days. Sulfotransferase activity also did not show the normal increase during the 3rd week of life and declined from 60% to 22%. Glucosyltransferase and lysosomal hydrolases in brain and ceramide galactosyltransferase in sciatic nerves appeared to be normal. These results suggest close similarities to the jimpy mutant mouse in which myelin deficiency is also inherited as an x-linked recessive trait.     

Zinc deficiency in wetland rice along a toposequence of hydromorphic soils in the Philippines

Summary Zinc deficiency in rice induced by prolonged water saturation of the soil is widespread in the Philippines. The poor drainage associated with the disorder is mostly due to upwelling of shallow artesian water in footslope areas and to ponding of water from continually flowing artesian wells common in such areas. One toposequence where no Zn deficiency occurred in the highest field and progressively severe Zn deficiency in successive lower fields, was studied in detail. With lower elevation soils showed an increase in organic matter and free carbonate, associated with increased upwelling and ponding of artesian water very high in dissolved Mg and Ca bicarbonate.     

X-linked dominant inheritance of partial phosphorylase kinase deficiency in mice

A new mouse strain, the V strain, with a partial deficiency of phosphorylase kinase has been established. The deficiency is caused by an X-linked dominant gene (Phk ^c ). Muscle extracts of homozygous and heterozygous females and hemizygous males have about 25% of the activity found in extracts of normal (C3H/HeHan) mice. This dominant phosphorylase kinase deficiency of the new V strain is different from that of the I-strain mice with the X-linked recessive deficiency of skeletal muscle phosphorylase kinase. The muscle extracts of V-strain and normal mice contain the same phosphorylase phosphatase activity of about 1 U/mg. Heart and liver extracts from V mice contained about 50% and 66%, respectively, of the phosphorylase kinase activity compared to that found in the same organs from the normal mice. The glycogen content of the skeletal muscle of the V strain was normal, i.e., 0.9 mg/g. Phosphorylase kinase was purified from the skeletal muscle of the V strain by (a) hydrophobic chromatography on methylamine Sepharose, (b) ammonium sulfate precipitation, and (c) gel filtration of Sepharose 4B. The enzyme has a similar structure to the normal murine and rabbit skeletal muscle enzyme, except that the proportion of the subunits differs. The molar ratio of the subunits of the V strain mice is (+)::=0.54:1:1.169, in comparison with that of the rabbit (+)::=1.1:1.0:1.0 and that of normal murine enzyme 0.9:1.0:0.7.This work was supported by the Minister für Wissenschaft und Forschung des Landes Nordrhein-Westfalen, West Germany and of the Fonds der Chemie, West Germany, and forms part of the md thesis of A. Vrbica.     

Evidence for two genetic complementation groups in pyruvate carboxylase-deficient human fibroblast cell lines

We have examined genetic complementation in pyruvate carboxylase deficiency by comparing the enzyme activity in polyethylene glycol-induced heterokaryons with that in unfused mixtures of fibroblasts from three affected children. Complementation, manifested as a three- to sevenfold increase in pyruvate carboxylase activity, was observed in fusions between a biotin-responsive multiple carboxylase (pyruvate carboxylase, propionyl CoA carboxylase, and -methylcrotonyl CoA carboxylase) deficient fibroblast line and two other lines deficient only in pyruvate carboxylase activity. Kinetic analysis of complementing pyruvate carboxylase deficient lines, measured by the rate of restoration of enzyme activity as a function of time, revealed that maximum restoration was achieved within 10–24 hr after fusion. This profile is similar to those observed for fusions between the multiple carboxylase deficient line and two lines deficient in propionyl CoA carboxylase activity that are known to represent different gene mutations. Although the patients with pyruvate carboxylase deficiency had similar clinical findings, our studies indicate that pyruvate carboxylase deficiency is genetically heterogeneous, with at least two distinct, probably intergenic, complementation groups.This work was supported by an NIH research grant (AM 25675) and an A. D. Williams research grant (6-48360). B. Wolf is the recipient of an NIH Research Career Development Award (AM 00677) and is aided by a Basil O'Connor Starter Research Grant from The National Foundation-March of Dimes (5-263). G. Feldman is the recipient of an NIH predoctoral training grant (GM 07492). This article is No. 100 from the Department of Human Genetics at the Medical College of Virginia.     

Mechanisms of 8‐aminoquinoline induced haemolytic toxicity in a G6PDd humanized mouse model

Primaquine (PQ) and Tafenoquine (TQ) are clinically important 8‐aminoquinolines (8‐AQ) used for radical cure treatment of P. vivax infection, known to target hepatic hypnozoites. 8‐AQs can trigger haemolytic anaemia in individuals with glucose‐6‐phosphate dehydrogenase deficiency (G6PDd), yet the mechanisms of haemolytic toxicity remain unknown. To address this issue, we used a humanized mouse model known to predict haemolytic toxicity responses in G6PDd human red blood cells (huRBCs). To evaluate the markers of eryptosis, huRBCs were isolated from mice 24–48 h post‐treatment and analysed for effects on phosphatidylserine (PS), intracellular reactive oxygen species (ROS) and autofluorescence. Urinalysis was performed to evaluate the occurrence of intravascular and extravascular haemolysis. Spleen and liver tissue harvested at 24 h and 5–7 days post‐treatment were stained for the presence of CD169+ macrophages, F4/80+ macrophages, Ter119+ mouse RBCs, glycophorin A+ huRBCs and murine reticulocytes (muRetics). G6PDd‐huRBCs from PQ/TQ treated mice showed increased markers for eryptosis as early as 24 h post‐treatment. This coincided with an early rise in levels of muRetics. Urinalysis revealed concurrent intravascular and extravascular haemolysis in response to PQ/TQ. Splenic CD169+ macrophages, present in all groups at day 1 post‐dosing were eliminated by days 5–7 in PQ/TQ treated mice only, while liver F4/80 macrophages and iron deposits increased. Collectively, our data suggest 8‐AQ treated G6PDd‐huRBCs have early physiological responses to treatment, including increased markers for eryptosis indicative of oxidative stress, resulting in extramedullary haematopoiesis and loss of splenic CD169+ macrophages, prompting the liver to act as the primary site of clearance.     

Reversible bacteriophage resistance by shedding the bacterial cell wall

Phages are highly abundant in the environment and pose a major threat for bacteria. Therefore, bacteria have evolved sophisticated defence systems to withstand phage attacks. Here, we describe a previously unknown mechanism by which mono- and diderm bacteria survive infection with diverse lytic phages. Phage exposure leads to a rapid and near-complete conversion of walled cells to a cell-wall-deficient state, which remains viable in osmoprotective conditions and can revert to the walled state. While shedding the cell wall dramatically reduces the number of progeny phages produced by the host, it does not always preclude phage infection. Altogether, these results show that the formation of cell-wall-deficient cells prevents complete eradication of the bacterial population and suggest that cell wall deficiency may potentially limit the efficacy of phage therapy, especially in highly osmotic environments or when used together with antibiotics that target the cell wall.     

Cd、Fe及其复合污染对烟草叶片几项生理指标的影响

本文通过盆栽和大田模拟试验,研究了Cd,Fe,Cd+Fe,Fe+Cd处理下烟草红花大金元(Nicotiana tabacurm L.)叶片的几项生理指标的变化情况。试验表明:烟草叶片叶绿素a、叶绿素b、叶绿素a+b含量均随Cd处理浓度的增加而下降,随Fe处理浓度的增加而上升。总烟碱含量随Cd处理浓度的增加(0—100ppm)而下降,Cd 150ppm时略有上升,随Fe处理浓度的增加而上升。蛋白质含量随Cd处理浓度的增加而上升,随Fe处理浓度的增加而下降。烟草的3个生理指标表明,Cd抑制Fe对生理指标的作用,Fe抑制Cd对它们的作用,Cd与Fe相互拮抗。Cd处理的烟草叶片过氧化物酶活性和细胞膜透性的测定结果表明,过氧化物酶活性和细胞膜透性均随Cd处理浓度的增加而上升。