Effect of the structure of the prosthetic group on the catalytic properties of prostaglandin H synthetase

The effect of vinyl groups of protohemin IX on its cofactor properties with respect to prostaglandin H synthetase has been studied. It was shown that substitution of ethyl groups or a hydrogen for vinyl groups affects neither binding of the prosthetic group to the apoenzyme nor catalytic properties of holo-prostaglandin H synthetase. Replacement of vinyl groups with bulkier substituents (hydroxyethyl or acetyl groups) decreases holoenzyme stability and catalytic activity. By comparison of the cofactor properties of protoporphyrin and hematoporphyrin macrocycles with different central ions (Fe3+, Mn2+, 2H+ in the case of protoporphyrin, and Fe3+, Mg2+, Cd2+ and Cu2+ in the case of hematoporphyrin), the presence of Fe3+ ions was shown to be mandatory for prostaglandin H synthetase activity. It was demonstrated that the cofactor structure modifications do not affect the holo-prostaglandin H synthetase inactivation rate constant in a reaction.     

Exploring the effect of oxygen-containing functional groups on the water-holding capacity of lignite

Graphene oxide with different degrees of oxidation was prepared and selected as a model compound of lignite to study quantitatively, using both experiment and theoretical calculation methods, the effect on water-holding capacity of oxygen-containing functional groups. The experimental results showed that graphite can be oxidized, and forms epoxy groups most easily, followed by hydroxyl and carboxyl groups. The prepared graphene oxide forms a membrane-state as a single layer structure, with an irregular surface. The water-holding capacity of lignite increased with the content of oxygen-containing functional groups. The influence on the configuration of water molecule clusters and binding energy of water molecules of different oxygen-containing functional groups was calculated by density functional theory. The calculation results indicated that the configuration of water molecule clusters was totally changed by oxygen-containing functional groups. The order of binding energy produced by oxygen-containing functional groups and water molecules was as follows: carboxyl > edge phenol hydroxyl >epoxy group. Finally, it can be concluded that the potential to form more hydrogen bonds is the key factor influencing the interaction energy between model compounds and water molecules.     

Changes in the composition of ionogenic groups of the wall of the lily pollen grain during germination

Changes in the composition of ionogenic groups of the polymeric matrix of the cell walls of lily (Lilium longiflorum Thunb.) pollen grains were studied during its activation at the early stages of pollen germination. In the cell walls isolated from nonactivated and activated pollen grains, four types of ionogenic groups were identified: amino groups, carboxylic groups of uronic acids, phenolic OH-groups. and groups with pK_a 7–8. During the early stages of germination, ionization constants of each type groups remained unchanged, but the quantitative composition of ionogenic groups in the intine changed. In this matrix, a decrease in the content of phenolic groups and demethylated carboxylic groups of uronic acids was detected. It is supposed that, at early stages of germination, the intine loses some part of acid pectins and some phenolic compounds.     

双氯芬酸钠栓和索利那新片治疗TURP术后OAB的临床疗效观察

目的:探讨双氯芬酸钠栓和索利那新治疗TURP术后OAB的临床疗效及预防性用药的必要性。方法:采取随机区组单盲的临床试验设计,将入选患者随机分为预防用药组和观察用药组。预防性用药组分别别使用双氯芬酸钠栓、索利那新片单药处理及两种药物联合处理,观察用药组在出现症状后按照预防性用药组的相同方法给药。结果:观察用药组中,双氯芬酸钠栓治疗的完全缓解率为50%,索利那新为81.8%,联合用药为90.5%,显著高于双氯芬酸钠栓(P0.05),各组不良反应的发生情况比较无显著性差异(P0.05)。预防用药组中,索利那新、双氯芬酸钠栓及联合用药分别可使TURP术后OAB的发生率降低45.0%、50.9%、53.8%.,三组之间比较无统计学差异(P0.05),且三组不良反应的发生情况比较无显著性差异(P0.05)。结论:双氯芬酸钠栓和索利那新治疗TURP术后OAB均安全有效,联合用药效果更佳,且预防性用药可以显著降低TURP术后OAB症状的发生率。     

The Effects of the Porosity of a Support and of Attachment on the Mode of Action of Immobilized Endopolygalacturonase

Endopolygalacturonase of Aspergillus sp. was immobilized by three different methods; viz. (a) via amino groups, (b) via carboxyl groups and (c) by means of epoxy groups to a nonporous microparticular silicon dioxide (Cabosil), functionalized by 3-(amino)-propyl groups and 3-(2′,3′-epoxypropoxy)-propyl groups, respectively. The conjugates were compared in their mode of action with corresponding immobilized preparations based on microporous ceramics. The binding via amino groups and via carboxyl groups lead, by itself, to changes in the mode of action of the enzyme, consisting of a decrease in randomness of glycosidic linkage splitting. The changes were greater in microporous support conjugates due to additional size-exclusion effects. The action pattern of endopolygalacturonase bound by means of epoxy groups was modulated exclusively by the porosity of the support, whereas the binding alone did not play any role.     

The Effects of the Porosity of a Support and of Attachment on the Mode of Action of Immobilized Endopolygalacturonase

Endopolygalacturonase of Aspergillus sp. was immobilized by three different methods; viz. (a) via amino groups, (b) via carboxyl groups and (c) by means of epoxy groups to a nonporous microparticular silicon dioxide (Cabosil), functionalized by 3-(amino)-propyl groups and 3-(2',3'-epoxypropoxy)-propyl groups, respectively. The conjugates were compared in their mode of action with corresponding immobilized preparations based on microporous ceramics. The binding via amino groups and via carboxyl groups lead, by itself, to changes in the mode of action of the enzyme, consisting of a decrease in randomness of glycosidic linkage splitting. The changes were greater in microporous support conjugates due to additional size-exclusion effects. The action pattern of endopolygalacturonase bound by means of epoxy groups was modulated exclusively by the porosity of the support, whereas the binding alone did not play any role.     

Ultrastructural observations on the cell surface of the intestinal epithelium of the nematode,Ascaris suum

Summary The intestinal epithelium of Ascaris suum consists of a single layer of tall columnar epithelial cells that rest on a thick basal membrane in contact with the pseudocoelomic cavity. Experiments were conducted on glutaraldehyde-fixed tissue to ascertain the nature of the electronegative charges associated with both the apical microvillar surface and basal membrane.A strong electronegative charge was demonstrated on the microvillar surface and basal membrane with ruthenium red and cationic ferritin staining. The ionic nature of ferritin binding was demonstrated with poly-L-lysine, a polycation that interacts with anionic groups on the membrane and thus blocks the subsequent binding of ferritin. Tissue thus treated was devoid of reaction product. Methylation with diazomethane completely abolished staining. Since the stronger acidic groups of sulfates or phosphates would not be protonated under the conditions employed in this study, and therefore susceptible to methylation, staining by ferritin is thought to be due to its interaction with carboxyl groups. Prior enzymatic treatment of tissue with neuraminidase or phospholipase C had no effect on subsequent ferritin binding. Tissue exposed to colloidal iron at various pH values showed maximal reactivity at a pH of 2.5 or above. Above pH 2.5, the dissociation of protons from free carboxyl groups of protein-bound amino-acid residues with pK's of 3.8 and 4.2 would be maximal, and the ionized carboxyl groups are then available to interact with iron micelles. These results suggest the presence of weaker acidic groups, such as the carboxyl groups of acidic amino acids or uronic acid residues. The stronger acidic groups of sialic acid and the esterified sulfate groups, if present, contribute only minimally to overall staining. These results demonstrate that a high electronegative charge density exists, despite the apparent lack of sialic acid. Staining is believed to be due to carboxyl groups of acidic amino acids and/or carboxyl groups or uronic acid residues.Part of this work was conducted at the Department of Zoology, Louisiana State University, Baton Rouge, Louisiana     

Localization of the carbohydrate residue of the S-layer glycoprotein from Clostridium thermohydrosulfuricum L111-69

The location of the covalently attached carbohydrate residue of the S-layer glycoprotein from Clostridium thermohydrosulfuricum L111-69 was determined by electron microscopical procedures after converting the hydroxyl groups of the carbohydrate chains into carboxyl groups by succinylation. The introduction of carboxyl groups was examined by labelling with polycationized ferritin (PCF; a net positively charged topographical marker for electron microscopy). Cyanogen bromide was used for activating vicinal hydroxyl groups of the carbohydrate chains which could then react with amino groups of amino carbonic acids or ferritin. The amount of covalently bound ferritin was determined by freeze-etching and UV-measurement. Both, succinylation experiments and covalent attachment of ferritin confirmed that at least a considerable portion of the carbohydrate residue must be located on the S-layer surface.     

DNA hyperreplication in the nuclei of Amoeba borokensis in the cell cycle]

The relative nuclear DNA contents were determined cytofluorometrically for several groups of synchronized Amoeba borokensis at the early and late interphase. In some groups of these amoebae the nuclear DNA content by the end of the interphase exceeded more than twice that measured 1 h after division, when DNA in amoebic nuclei already being synthesized. This means that the extra DNA was synthesized in the nuclei of amoebae of these particular groups. In other amoebic groups the nuclear DNA content checked at the end of the interphase did not exceed the doubled 1 h level. Thus, in these amoebae the quantity of the synthesized extra DNA was less than that in the former groups, or the extra DNA was not synthesized at all.     

A comparison of the toxicity of certain dyestuffs to the conidia of Fusarium culmorum

The toxicity of a number of dyestuffs to the spores of Fusarium culmorum and Cercosporella herpotrichoides was determined by the slide-germination technique. No attempt was made to distinguish between fungistatic and fungicidal activity.
The toxicity of basic dyestuffs was unaffected by the acid radicle associated with the dye base.
The high toxicity to Fusarium culmorum of malachite green dye base was reduced weight for weight and mole for mole by substitution of ethyl, propyl or butyl groups for methyl groups.
The reduction of malachite green to malachite green leuco base removed toxicity.
The substitution of amino groups and alkylated amino groups in benzene nuclei of triphenyl methane increased toxicity, whereas acid groups reduced toxicity. Sulphonation and carboxylation reduced toxicity to vanishing point.
Alkylation of amino groups increased, but alkylation of benzene nuclei did not affect toxicity appreciably.
When the central carbon atom of the triphenyl methane dyestuffs was replaced by nitrogen (e.g. Bindschedler's green) the diphenyl ammonium compounds were less toxic than the corresponding triphenyl-methane compounds.
The prevention of rotation of the aminated benzene rings by bridging, in the o -position to central atom, with O or N, and so obtaining a planar molecule only slightly affected toxicity.
Certain acid dyes stimulated fungal growth.
The toxicity of the basic dyestuffs seems to depend not on one specific part but on the molecule as a whole, and within certain limits the structure may be varied without pronounced changes in toxicity.     

The reactivity of the thiol groups of the adenosine triphosphatase of sarcoplasmic reticulum and their location on tryptic fragments of the molecule.

The ATPase (adenosine triphosphatase) from sarcoplasmic reticulum contains 20 thiol groups/115000 daltons, measured by using either N-ethyl[(14)C]maleimide or 5,5'-dithiobis-(2-nitrobenzoate) in sodium dodecyl sulphate. After reduction there were 26 thiol groups, in good agreement with 26.5 residues of cysteic acid found by amino acid analysis. The difference between this and the 20 residues measured before reduction implies the presence of three disulphide residues. The same number of disulphide residues was found by direct measurement. Three to six fewer thiol groups were found in preparations made in the absence of dithiothreitol. The missing residues were accounted for as cysteic acid. The distribution of disulphide bonds and of exposed and buried thiol groups among the tryptic fragments of the molecule was measured after labelling with N-ethyl[(14)C]-maleimide. The disulphides were confined to fragment B (mol.wt. 55000), whereas several thiol groups were present on each of the fragments (A, B, A(1) and A(2)). The kinetics of the reaction of the ATPase with 5,5'-dithiobis-(2-nitrobenzoate) showed that four or five of the thiol groups were unreactive in the absence of detergent and that 13 of the remainder reacted with a single first-order rate constant. In the presence of ATP and Ca(2+) the reaction rate of all but two groups of this class was uniformly decreased. In the presence or absence of ATP and Ca(2+) the rate constant for inactivation was close to the rate constant for this class, but was not identical with it. No selective protection of a specific active-site-thiol group was observed. Parallel experiments with sarcoplasmic reticulum gave similar results, except that the reaction rates were a little lower and there were two more buried groups. Solution of ATPase of sarcoplasmic reticulum in detergent greatly increased the reactivity of all thiol groups. The effects of low concentrations of deoxycholate were reversible. EGTA or low concentrations (0.02mm) of Ca(2+) of Mg(2+) had very little effect on the reactivity.     

兔坐骨神经火器伤模型的建立与选择

目的　建立并评价兔坐骨神经火器伤模型。方法　实验用大耳白兔 2 6只根据致伤钢珠质量和装药量随机分为 5组 ,其中A、B、C组用 0 3 8g钢珠 ,装药量分别为 0 3 5g、0 65g和 1 2 0g ;D、E组用 1 0 3g钢珠 ,装药量分别为 0 3 5g和 0 65g。致伤靶点为兔右后肢外侧坐骨神经体表投影线中点。观察指标有致伤钢珠入出口速度、传递能量、伤道入出口大小、骨折与血管伤发生情况、坐骨神经干损伤长度、坐骨神经病理变化。结果　A组 ( 62 1 6m s)和D组 ( 5 94 6m s)的致伤速度偏低 ,C组的 ( 12 99 8m s)偏高 ,B组 ( 93 9 8m s)和E组 ( 816 1m s)的较接近。A组 ( 18 7J)和D组 ( 4 4 2J)的传递能量较小 ,未合并骨折和血管伤 ,伤情较轻。C组 ( 183 2J)和E组 ( 165 2J)的传递能量较大 ,合并骨折和血管伤机率高 ,伤情较重。B组的传递能量适中 ( 80 0J) ,合并骨折和血管伤机率小。B、C、E组均可见典型坐骨神经火器伤病理改变 ,A、D组坐骨神经损伤程度较B、C、E组的轻。结论　在B组致伤条件下 ,既易造成典型的兔坐骨神经火器伤 ,也适于周围神经火器伤需长期观察的实验要求。     

An anatomic atlas of the medulla oblongata of the adult goat.

An anatomic atlas of the goat brain stem was developed for use in studies that analyze medullary neuronal groups, and factors that influence variability in the location of neuronal groups were determined. The medullas of 31 adult goats (weight, 17-88 kg) were fixed, harvested, frozen, serially sectioned, stained with 0.5% neutral red, and examined with a light microscope. Obex, the point at which the central canal opens into the fourth ventricle, was taken as the zero reference point from which the rostrocaudal and mediolateral coordinates of medullary neuronal groups were determined, whereas dorsoventral coordinates were calculated from the medullary surface. Histological variations with goat body weight were quantified, and linear regression analysis provided adjustment factors for weight in all three dimensions. Similar analysis of percentage of shrinkage on fixation and processing provided adjustment factors for precise coordinates of medullary neuronal groups. For accurate location of neuronal groups, body weight and histological procedure should be taken into account. The present study provided adjustment factors for body weight and standard histological processing to locate most major medullary neuronal groups in the adult goat.     

Regulatory mechanisms of cellular respiration; the role of soluble sulfhydryl groups as shown by the effect of sulfhydryl reagents on the respiration of sea urchin sperm

Oxidizing agents of sulfhydryl groups such as iodosobenzoate, alkylating agents such as iodoacetamide, and mercaptide-forming agents such as cadmium chloride, mercuric chloride, p-chloromercuribenzoate, sodium arsenite, and p-carboxyphenylarsine oxide, added in small concentrations to a suspension of sea urchin sperm produced an increase in respiration. When the concentration was increased there was an inhibition. These effects are explained by postulating the presence in the cells of two kinds of sulfhydryl groups: soluble sulfhydryl groups, which regulate cellular respiration, and fixed sulfhydryl groups, present in the protein moiety of enzymes. Small concentrations of sulfhydryl reagents combine only with the first, thus producing an increase in respiration; when the concentration is increased, the fixed sulfhydryl groups are also attacked and inhibition of respiration is the consequence. Other inhibitors of cell respiration, such as cyanide and urethanes, which do not combine with -SH groups, did not stimulate respiration in small concentration.     

Increase of the molecular rigidity of the protein conformation in the intestinal brush-border membranes by lipid peroxidation

The effect of lipid peroxidation on the protein conformation of the porcine intestinal brush-border membranes was studied using a fluorogenic thiol reagent, N-[7-dimethylamino-4-methylcoumarinyl]maleimide (DACM). By a kinetic analysis of the reaction of the membranes with DACM, it was shown that the reaction rate of the SH groups (SHf) of the membrane proteins, whose reaction with the dye is very fast, decreases in proportion to the extent of thiobarbituric acid-reactive substance formation. The difference in the rate of the reaction of the SHf groups for DACM between the control and peroxidized membranes completely disappeared after denaturation of the proteins by treatment with guanidine hydrochloride. The reaction of DACM with the SHf groups of the control membranes accelerated when the temperature was increased with an apparent transition temperature between 25 degrees C and 30 degrees C. On the other hand, no transition was observed in the peroxidized membranes over the temperature range 20-43 degrees C. These results suggest that the conformation around the SHf groups of the proteins in the peroxidized membranes is apparently different from that in the control membranes. A modification of the conformation around the SH groups in the membrane proteins associated with lipid peroxidation was further demonstrated by finding that the quenching efficiency of the fluorescence of the DACM-labeled membranes by Tl+ was markedly decreased after lipid peroxidation. Based on these results, changes in the protein conformation of the porcine intestinal brush-border membranes by lipid peroxidation are discussed.     

Influence of the Dietary Protein Deficiency on the Activities of Ribosomes and Polysome Patterns in Muscle and Liver of Rats

A group of rats weighing about 120 g were killed at the beginning of the experiment and after 10 days on the 20% casein diet (C-0 and C-10 groups), and another group of rats were killed after 1, 2 and 10 days on the protein-free diet (PF-1, PF-2 and PF-10 groups). From muscle and the liver of each group ribosomes were prepared, and the protein synthesis activity and the polysome patterns were investigated. The activity of polysome fractionated into each size was also measured.

Muscle ribosome activity in PF-1, PF-2 and PF-10 groups decreased to about 60%, 40% and 40% of that in C groups, respectively, and this decrease was due to a fall in activity of polysome itself rather than disaggregation of polysome. Liver ribosome activity in PF-1, PF-2 and PF-10 groups were reduced to about 95%, 90% and 65% of that in C groups, respectively. These alterations in PF-1 and PF-2 groups seemed to be in part related to changes in polysome pattern, whereas ribosome activity in PF-10 group was reduced without changes in polysome pattern.     

A one-step liquid-chromatographic technique for the estimation of the deoxythymidine-5'-triphosphatase in human serum

Deoxythymidine-5'-triphosphatase (dTTPase) from human serum was separated by DEAE-cellulose chromatography from unspecific hydrolases in a single step. The method was adapted to a microscale and the enzymatic activity was determined for five different groups of patients including epithelial carcinoma, leukemia and three control groups. The result is that the purified dTTPase preparations of these groups reflect the situation obtained in the whole serum.     

不同地区斑点叉尾鮰种内遗传多样性分析

采用RAPD方法对来源于美国三个地区的30条斑点叉尾鮰进行遗传多样性的分析。对60个随机引物进行筛选,选出能够区分地区内以及地区间种群遗传多态性的引物。通过统计学的方法计算分析得出三个地区各自种群内遗传相似率,德克萨斯州为82.52%,阿纳巴马州为80.46%,密西西比州为83.71%。对地区之间种群的遗传分化情况进行t检测分析得出三个地区之间斑点叉尾鮰种群的遗传特性各不相同,其中差异最大的是阿纳巴马州和密西西比州,差异最小的是德克萨斯州和密西西比州。     

Effect of sulfhydryl compounds on the automatism of the pacemakers]

Experiments were performed on the isolated rabbit hearts and also on the hearts with complete atrioventricular block; a study was made of the effect of an excess or deficiency of the sulfhydryl groups on the automatism of the cardiac pace makers. Unithiol and cysteine in concentrations of 1-10(-6)-1-10(-4) g/ml were used as donors of sulfhydryl groups; deficiency of these groups was induced by the alloxan administration in concentrations of 1-10(-5)-5-10(-5) g/ml. Changes in the sulfhydryl group content produced no marked effect on the automatism of the synoatrial node. An excess of sulfhydryl groups promoted poststimulation depression of the automatism of the potential pace makers of the cardiac ventricles and could lead to the origination of Luciani's periods. On the contrary, in case of a deficiency of the sulfhydryl groups there was a sharp elevation of the automatism of the ventricular pace makers, the atrioventricular conduction became disturbed, and the poststimulation depression of the automatism became considerably diminished. Disturbances of the cardiac activity caused by the sulfhydryl group deficiency were completely eliminated by unithiol or cysteine.     

A method for the estimation of thiol esters.

1. A method is described for the estimation of thiol ester groups. The thiol ester is converted into the corresponding thiol by reaction with ammonia; the thiol is then titrated amperometrically with mercuric chloride. 2. The method may be used in the presence of SH and S.S groups. The SH groups are titrated at pH3 in the presence of excess of chloride; under these conditions thiol esters do not react with mercuric chloride. Thiol ester plus thiol is then estimated by titration after reaction with ammonia. Finally, titration after reaction with ammonia and sulphite gives the thiol ester plus thiol plus disulphide. 3. The procedure has been applied to glyceraldehyde phosphate dehydrogenase. The enzyme was found to contain 15-16 SH groups/mol. and no S.S groups. After reaction with acetyl phosphate 1.8-3.5 thiol ester groups were detected, the number depending on the conditions of acetylation. In the absence of bound NAD, the number of thiol ester groups formed was 1.8/mol., although a value of 2.9 labile acetyl groups/mol. was given by the method of Lipmann & Tuttle (1945). The presence of thiol ester groups in the S-(d-3-phosphoglyceryl)-enzyme was also demonstrated.