Requirement of trypsin inhibitor for measurement of 3-hydroxy-3-methylglutaryl coenzyme A reductase in intestinal mucosa of rat

A method was developed for the determination of 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in the microsomal fraction of crypt cells and villi of rat intestinal mucosa. Addition of trypsin inhibitor to homogenizing and incubation media at a proper concentration appeared inevitable for measurement of the activity of the villi fraction. The reductase in crypt cells was also slightly enhanced by the addition of the inhibitor. Using this technique, the enzyme activity in villi was found to be as active as the crypt cell fraction. Since other types of protease inhibitors were not necessarily effective, it was suggested that specific enzyme(s) inactivates the mucosal reductase in the course of measurement.     

Central neural peptides and catecholamines in spontaneous and DOCA/salt hypertension

Hypothalamic and neurophypophyseal levels of catecholamines and peptides were measured in spontaneous and deoxycorticosterone (DOCA)/salt hypertension. Catecholamines, norepinephrine, epinephrine and dopamine were measured by electrochemical detection while the peptides, vasopressin, oxytocin, luteinizing hormone-releasing hormone (LHRH), the enkephalins and somatostatin (SRIF) were measured by radioimmunoassay. Blood pressure was significantly elevated in both groups as compared to their controls. Marked changes in central neural peptides were observed in the SHR, while no differences were seen in DOCA/salt hypertension. Hypothalamic vasopressin, oxytocin, LHRH and SRIF were significantly decreased. In the posterior pituitary, enkephalins were increased twofold in the SHR. With regard to catecholamines, there was no change in hypothalamic content. However, a dramatic decrease in neurohypophyseal dopamine was observed in SHR. Plasma levels of vasopressin were significantly elevated in both types of hypertension while oxytocin was increased only in the DOCA/salt model. These result show that (1) a wide spectrum of neuroendocrine changes are associated with genetic hypertension, (2) there are CNS differences between DOCA/salt and spontaneous hypertension, and (3) central aminergic changes may be involved in th neuroendocrine alterations seen in the SHR.     

Rat and human embryo and post-natal sera contain a potent endogenous competitor of estrogen-rat alpha-fetoprotein interactions

A highly active inhibitor of the binding of estrone and estradiol-17β to rat alpha-fetoprotein is demonstrated for the first time in embryo, immature and adult rat sera as well as in fetal and adult human sera. The competitive character and the narrow specificity of this inhibition effect is shown. The major compound responsible for this activity is isolated by successive column Sephadex LH₂₀ and thin layer chromatography : it is characterized as a nonpolar, nonphenolic, dialysable and thermostable substance, unreactive towards anti-estrone and anti-estradiol-17β anti-bodies. The possible biological role of an endogenous non-estrogen ligand of rodent fetoproteins is discussed.     

Dependence of the early steps of chlorophyll(ide) synthesis on respiration in dark-grown Euglena gracilis

Protochlorophyll(ide) disappearance and chlorophyll(ide) accumulation, in dark-grown Euglena, promoted by series of actinic light flashes, have been followed by in vivo fluorescence measurements. The data show that chlorophyll(ide) accumulation is biphasic, i.e., there is an initial rapid phase followed by a slower linear phase. The linear phase is highly dependent on flash frequency and on cell respiration whereas the initial phase is much less affected by these factors. It is concluded that dark-grown cells contain a limited pool of phototransformable protochlorophyll(ide); once this pool is exhausted, its reformation and/or the synthesis of some unknown metabolite necessary for the photoreduction appears to be dependent on respiration.     

Root-zone temperature and salinity: Interacting effects on tillering,growth and element concentration in barley

The effects of root-zone salinity (0, 30, and 60 mmol L^–1 of NaCl) and root-zone temperature (10, 15, 20, and 25°C) and their interactions on the number of tillers, total dry matter production, and the concentration of nutrients in the roots and tops of barley (Hordeum vulgare L.) were studied. Experiments were conducted in growth chambers (day/night photoperiod of 16/8 h and constant air temperature of 20°C) and under water-culture conditions. Salinity and root temperature affected all the parameters tested. Interactions between salinity and temperature were significant (p<0.05) for the number of tillers, growth of tops and roots, and the concentration of Na, K, P in the tops and the concentration of P in the roots. Maximum number of tillers and the highest dry matter were produced when the root temperature was at the intermediate levels of 15 to 20°C. Effect of salinity on most parameters tested strongly depended on the prevailing root temperature. For example, at root temperature of 10°C addition of 30 mmol L^–1 NaCl to the nutrient solution stimulated the growth of barley roots; at root temperature of 25°C, however, the same NaCl concentration inhibited the root growth. At 60 mmol L^–1, root and shoot growth were maximum when root temperature was kept at the intermediate level of 15°C; most inhibition of salinity occurred at both low (10°C) and high (25°C) root temperatures. As the root temperature was raised from 10 to 25°C, the concentration of Na generally decreased in the tops and increased in the roots. At a given Na concentration in the tops or in the roots, respective growth of tops or roots was much less inhibited if the roots were grown at 15–20°C. It is concluded that the tolerance of barley plant to NaCl salinity of the rooting media appears to be altered by the root temperature and is highest if the root temperature is kept at 15 to 20°C.     

Spatio-temporal tumour model for analysis and mechanism of action of intracellular drug accumulation

We have developed a one-dimensional tumour simulator to describe the biodistribution of chemotherapeutic drugs to a tumoral lesion and the tumour cell’s response to therapy. A three-compartment model is used for drug dynamics within the tumour. The first compartment represents the extracellular space in which cells move, the second corresponds to the intracellular fluid space (including cell membrane) which is in direct equilibrium with the extracellular space, and the third is a non-exchangeable compartment that represents sequestered drug which is trapped in the nucleus to damage the cellular DNA, directly triggering cell death. Analytical and numerical techniques (Finite Element Method) are used to describe the tumour’s response to therapy and the effect of parameter variation on the drug concentration profiles in the three compartments.     

Identification of salt-stress responsive genes in rice (Oryza sativa L.) by cDNA array

To identify salt stress-responsive genes, we constructed a cDNA library with the salttolerant rice cultivar, Lansheng. About 15000 plasmids were extracted and dotted on filters with Biomeck 2000 HDRT system or by hand. Thirty genes were identified to display altered expression levels responding to 150 mmol/L NaCl. Among them eighteen genes were up-regulated and the remainders downregulated. Twenty-seven genes have their homologous genes in GenBank Databases. The expression of twelve genes was studied by Northern analysis. Based on the functions, these genes can be classified into five categories, including photosynthesis-related gene, transportrelated gene, metabolismrelated gene, stress-or resistancerelated gene and the others with various functions. The results showed that salt stress influenced many aspects of rice growth. Some of these genes may play important roles in plant salt tolerance.     

Nitrogen-molybdenum-manganese co-fertilization reduces nitrate accumulation and enhances spinach (Spinacia oleracea L.) yield and its quality

Spinach (Spinacia oleracea L.) is considered a nitrogen (N) intensive plant with high nitrate (NO₃^?) accumulation in its leaves. The current study via a two-year field trial introduced an approach by combining N fertilization from different sources (e.g., ammonium nitrate; 33.5 % N, and urea; 48 % N) at different rates (180, and 360 kg N ha^?1) with the foliar spraying of molybdenum (Mo) as sodium molybdate, and/or manganese (Mn) as manganese sulphate at rates of 50 and 100 mgL^?1 of each or with a mixture of Mo and Mn at rates of 50 and 50 mg L^?1, respectively on growth, chemical constituents, and NO₃^? accumulation in spinach leaves. Our findings revealed that the highest rate of N fertilization (360 kg N ha^?1) significantly increased most of the measured parameters e.g., plant length, fresh and dry weight plant^?1, number of leaves plant^?1, leaf area plant^?1, leaf pigments (chlorophyll a, b and carotenoids), nutrients (N, P, K, Fe, Mn, Zn), total soluble carbohydrates, protein content, net assimilation rate, and NO₃^? accumulation, but decreased leaf area ratio and relative growth rate. Moreover, plants received urea-N fertilizer gave the highest values of all previous attributes when compared with ammonium nitrate –N fertilizers, and the lowest values of NO₃^? accumulation. The co-fertilization of N-Mo-Mn gave the highest values in all studied attributes and the lowest NO₃^? accumulation. The best treatment was recorded under the treatment of 360 kg N-urea ha^?1 in parallel with the combined foliar application of Mo and Mn (50 + 50 mg L^?1). Our findings proposed that the co-fertilization of N-Mo-Mn could enhance spinach yield and its quality, while reducing NO₃^? accumulation in leaves, resulting agronomical, environmental and economic benefits.