Seasonal variation of xanthine oxidoreductase activity in the digestive gland cells of the mussel Mytilus galloprovincialis: a biochemical, histochemical and immunochemical study

The activity and the tissue distribution of the oxygen radical producing enzyme xanthine oxidoreductase (XOR) were measured in the digestive gland of the common marine mussel Mytilus galloprovincialis Lmk along an annual cycle. No xanthine oxidase (XOX) activity could be measured, the enzyme only displaying xanthine dehydrogenase (XDH) activity in all the cases. This is interpreted as a mechanism to avoid the harmful effects of the oxygen radicals that would be produced by XOX during periods following anoxic conditions at low tide. The highest XDH activities coincided with the late spring/early summer months, the activity maxima being recorded from May to July. Histochemically XOR activity was very pronounced in duct and stomach epithelial cells as well as in the surrounding connective tissue and hemolymph vessels, the activity increasing towards the summer months. These seasonal variations in XDH or XOR activities are possibly linked to hormonal changes governing the reproductive cycle and to changes in food availability. The localization of the protein in the connective tissue lining the hemolymph vessels was confirmed immunohistochemically using a polyclonal antibody against rat liver protein that cross-reacted specifically with a polypeptide of 150 kDa of molecular mass in homogenates of the digestive gland. This polypeptide was linked to cytosolic fractions isolated by differential centrifugation from mussel digestive glands. In paraffin sections the antibody labeled the digestive cells of digestive tubules, as well as the connective tissue surrounding the hemolymph vessels, gonadal follicles, digestive epithelia and certain protozoan parasites. Taken together our results suggest that in the digestive gland of bivalve molluscs XOR is involved in the metabolism of purines and in the scavenging of oxygen free radicals.     

油松含树脂细胞组织化学和细胞化学研究

油松茎端的皮层薄壁组织和幼茎木薄壁组织中具含树脂细胞分布,在光学显微镜下观察,硫酸奈尔兰染色的切片,发现树脂滴分布于细胞质中,在电子显微镜下观察狐染色切片,发现树脂滴分布于细胞的线粒体,高尔基体,内质网,核膜和小液泡中,同时也分布于质膜两边。从形态演化分析含树脂细胞在树脂分泌结构中属原始类型,由它演化为树脂道。     

急性肾功能衰竭对肝损害的实验研究

探讨急性肾功能衰竭 (以下简称为 ARF)时尿毒症毒素对肝的损害情况 ,为临床防治提供形态学依据。Wistar大白鼠 32只 ,随机分成正常对照组 ,14只 ;双输尿管结扎结扎组 ,18只。模型建成后 2 3- 37小时内将动物处死 ,取肝组织作NOS、 MAO、 SDH、 L DH、 Ch E、 ATPase、 ACP等项酶组织化学显色及超微结构观察。结果显示 :实验组 SDH、 Mg2 + -ATPase 的活性均由正常的强阳性 ( )下降为阳性 (+) ;MAO、 Ch E的活性均由正常的中等阳性 ( )下降为阳性(+) ;ACP、 L DH的活性均由阳性 (+)增强到中等阳性 ( ) ;NOS活性由正常的弱阳性 (± )增强为阳性 (+) ;肝细胞器结构受损。结果表明 :ARF的尿毒素的大量淤积通过影响肝脏酶的活性来干扰和抑制糖代谢、三羧酸循环、蛋白质合成及解毒等功能 ,并导致肝损害发生。 2用改良的 NADPH-黄递酶法可显示肝组织 NOS的存在 ,该法操作简便 ,经济 ,特别适用于同时需要留取活组织做其它检查者。当输尿管结扎时 ,NOS的确切作用有待于进一步探讨     

青藤与毛青藤茎中青藤碱积累的组织化学研究

青藤及毛青藤茎中青藤碱积累的组织化学研究表明,青藤碱主要分布在皮层和接近维管形成层的射线细胞中,在髓鞘及细胞壁已木质化加厚的射线细胞内也有少量青藤碱分布,且木化的细胞壁中浸透着青藤碱。     

在沉积物暴露条件下2种海洋贝类对Cu、Pb 的富集动力学研究

首次通过构建海水-底泥-生物体体系,应用半静态双箱动力学模型室内模拟了沉积物暴露条件下文蛤(Meretrix meretrix)和缢蛏(Sinonovacula constricta)对Cu、Pb的生物富集实验,通过对富集与排出过程中文蛤体内重金属污染物的动态监测和对富集与排出过程监测结果的非线性拟合,得到了文蛤和缢蛏富集重金属的吸收速率常数K1、排出速率常数K2、生物富集因子BCF、富集平衡浓度CAmax、生物学半衰期B1/2等动力学参数。拟合结果得到的Cu、Pb各动力学参数分别为:文蛤K1为4.63—69.41,K2为0.05—0.08,CAmax为9.83—40.57,BCF为60.77—1444.56,B1/2为9.09—14.43;缢蛏K1为4.27—122.92。K2为0.05—0.07。CAmax为5.12—32.84。BCF为65.68—2112.02。B1/2为9.81—14.15。对模型的拟合优度检验结果显示,在沉积物暴露条件下文蛤和缢蛏对重金属Cu、Pb的生物富集数据符合双箱模型,模型的拟合优度良好。比较结果得出:吸收速率常数K1及生物富集因子BCF均随着外部水体金属暴露浓度的增大而减小;CAmax均随着外部水体金属暴露浓度的增大而增大;文蛤和缢蛏对Cu富集能力大于Pb;Cu在文蛤和缢蛏体内的生物学半衰期B1/2大于Pb;理论平衡状态下文蛤和缢蛏体内Cu、Pb的含量CAmax随着外部水体中金属暴露浓度的增大而增大,且呈显著正相关,实验结果表明在沉积物暴露条件下双箱动力学模型在一定条件下是可以应用于文蛤和缢蛏的富集动力学研究的,但仍需要进一步开展在不同条件下实验研究分析。     

氧自由基在应激性胃溃疡中的发病学意义

本工作研究了氧自由基在大鼠冷冻束缚应激性胃溃疡中的发病学意义。实验结果如下:(1)以超氧自由基清除剂超氧化物歧化酶(SOD)或羟自由基清除剂二甲亚砜和甘露醇预先处理大鼠,均可显著地减轻胃粘膜损伤;(2)应激时,胃粘膜内的脂质过氧化分解产物丙二醛的含量显著升高;(3)组织化学的研究显示,胃粘膜层含有丰富的黄嘌呤氧化酶,其活性在应激时明显升高,预先用别嘌呤醇处理大鼠以抑制黄嘌呤氧化酶的活性,可使胃粘膜损伤显著减轻。上述结果提示,氧自由基是应激性胃溃疡的重要致病因子,而黄嘌呤氧化酶活性的升高似可能为应激时氧自由基生成增加的重要原因。     

实验性脾虚证大鼠结肠和盲肠组织化学、免疫组织化学研究

成年雄性Wistar大鼠60只,分正常对照组;脾虚组;自然恢复组和四君子汤治疗组。四组动物取结肠和盲肠分别进行H-E;酶组织化学和5-羟色胺细胞PAP免疫组织化学观察。结果表明,脾虚组结肠和盲肠粘膜上皮的杯状细胞较对照组明显增多,淋巴细胞及淋巴集结也增多。固有层毛细血管充血并有炎症细胞的增多。上皮细胞和腺细胞琥珀酸脱氢酶(SDH)减弱,乳酸脱氢酶(LDH)增强;结肠上皮细胞和腺细胞碱性磷酸酶(AIP)和三磷酸腺苷酶(ATPase)增强,而盲肠上皮细胞和腺细胞AIP和ATPase减弱。结肠和盲肠5-HT细胞免疫组化反应减弱。这些结果表明肠上皮细胞酶的变化和5-HT细胞分泌活性的变化与脾虚证的发生密切相关,可能是导致脾虚证原因之一。经四君子汤治疗后以上各项指标均比自然恢复组更接近于正常对照组,说明此药对消化道粘膜上皮酶活性和5-HT细胞分泌活性恢复正常有明显效果,从而起治疗作用。     

光对水稻(Oryza sativa)幼苗乙醇酸氧化酶活性的影响

水稻黄化幼苗用白光照射3～5小时后可诱导出乙醇酸氧化酶活性。绿色水稻幼苗在黑暗中乙醇酸氧化酶活性逐渐下降以至消失,再给以照光又恢复酶活性。亚胺环己酮对光的诱导及再诱导均有抑制作用。在黑暗中真空渗入乙醇酸于黄化幼苗可诱导出乙醇酸氧化酶活性,渗入乙醇酸加FMN能使酶活性迅速增强。弱的白光以及红、绿,蓝光均有诱导作用。因此认为光的诱导作用是使黄化幼苗形成乙醇酸——作为诱导物,以诱导乙醇酸氧化酶的新合成。光也可以加速叶组织内FMN的合成,从而提高乙醇酸氧化酶的活性。     

间苯二酚、水杨酸对绿豆下胚轴不定根形成的作用

２０—１００ｍｇＬ（－１）间苯二酚能明显地促进绿豆下胚轴不定根的形成,与２０ｍｇＬ（－１）ＩＢＡ混合处理具加成效应,其作用在于降低生根初期ＩＡＡ氧化酶和多酚氧化酶活性．１０—１００ｍｇＬ（－１）水杨酸抑制下胚轴不定根的形成,随处理浓度的加大,对生根数目、生根范围和根重的抑制作用增加．水杨酸处理后１－３ｄ,能提高ＩＡＡ氧化酶和多酚氧化酶的活性．     

间苯二酚、水杨酸对绿豆下胚轴不定根形成的作用

２０—１００ｍｇＬ（－１）间苯二酚能明显地促进绿豆下胚轴不定根的形成,与２０ｍｇＬ（－１）ＩＢＡ混合处理具加成效应,其作用在于降低生根初期ＩＡＡ氧化酶和多酚氧化酶活性．１０—１００ｍｇＬ（－１）水杨酸抑制下胚轴不定根的形成,随处理浓度的加大,对生根数目、生根范围和根重的抑制作用增加．水杨酸处理后１－３ｄ,能提高ＩＡＡ氧化酶和多酚氧化酶的活性．     

MULTIPLE ORIGINS OF GENDER-ASSOCIATED MITOCHONDRIAL DNA LINEAGES IN BIVALVES (MOLLUSCA: BIVALVIA)

Previous studies have shown that marine mussels (genus Mytilus) and a freshwater mussel (Pyganodon grandis) contain two distinct gender-associated mitotypes, which is a characteristic feature of the phenomenon of doubly uniparental inheritance (DUI) of mitochondrial DNA (mtDNA). Here we present evidence for the presence of distinct male (M) and female (F) mitotypes in three other bivalve species, the mytilid Geukensia demissa, and the unionid species P. fragilis and Fusconaia flava. Nucleotide sequences of a segment of the COI gene from the M and F mitotypes from each of the three mytilid species (M. edulis, M. trossulus, G. demissa) and three unionid species (P. grandis, P. fragilis, F. flava) were used for phylogenetic analysis. The analysis suggests three independent origins of M and F mitotypes for the six species examined; one for the three unionid species, one for the two Mytilus species, and one for Geukensia. The first of these F/M divergence events, while of uncertain age, predates the divergence of the two unionid genera and is likely older than either of the two F/M divergence events in the mytilid taxa. The most parsimonious explanation of multiple F/M divergence events is that they represent independent origins of DUI. Another possibility is that, in a given taxon, an F or M mitotype assumes the role of the opposite mitotype (by virtue of a mechanism that remains to be clarified) and subsequently was fixed within its new gender. The fixation of a mtDNA lineage derived from a mitotype of switched function would reset the divergence of the gender-associated lineages to zero, thereby mimicking a de novo split of F and M lineages from a preexisting mtDNA genome that was not gender specific. Further broad-scale taxonomic studies of the occurrence of distinct M and F mitotypes may allow for the evaluation of the latter hypothesis.     

草鱼不同组织胆汁酸组成分析

正胆汁酸是由胆固醇通过一系列酶促反应在肝细胞中合成的,根据侧链羟基数目的多少胆汁酸分为初级胆汁酸和次级胆汁酸。初级胆汁酸随食物到达肠道,在肠道微生物的作用下脱羟基形成次级胆汁酸~([1])。初级胆汁酸和次级胆汁酸都可与牛磺酸或者甘氨酸结合形成结合型胆汁酸。肝脏中合成的以及经由肠肝循环回到肝脏的胆汁酸都被储存在胆囊中。胆汁酸的主要生理功能是帮助脂溶性营养物质的消化吸收,高脂肪的摄入刺激胆汁酸     

酸雨对重庆几种土壤中元素释放的影响

本文通过一个严格的实验设计,研究了荒地,人工马尾松林,樟树林,公园耕作土等４种土壤的自然土柱在受到完全根据天然降水而配制的不同酸度的人工模拟酸寸在严格的人工控制下的淋洗后,淋洗液的ＰＨ值变化和各种物质元素浓度的变化规律,表明了重庆南山的主要土壤都存在一定的抗酸化能力,在这个临界值范围内,土壤溶液的ＰＨ值及其中物质的元素浓度都相对稳定;而在此范围以外,土壤溶液ＰＨ值急剧下降。Ａｌ＾３＋、Ｃａ＾２＋、     

酶标抗原火箭免疫电泳法及其在蛇毒抗原成分含量测定中的应用

为解决目前蛇毒成份检测困难以及寻找一种可用于蛇伤快速鉴别诊断的方法,本研究建立了酶标抗原火箭免疫电泳法。本法结合了酶标法灵敏度高和火箭电泳操作简单的特点,对待测的同种抗原标记辣根过氧化物酶(酶标抗原),检测时把微量的酶标记抗原掺入被测样品中,在含多价抗血清的免疫电泳板进行电泳。最后用辣根过氧化物底物3.3′-二氨基联苯胺在板上直接显色,显色后观察到的免疫沉淀线高度与被测抗原含量呈正相关(r=+0.98)。采用本方法检测眼镜蛇毒细胞毒素和限镜王蛇毒 L-氨基酶氧化酶,细胞毒素的最低检测浓度为110ng/ml,氨基酸氧化酶为60ng/ml。比单向火箭免疫电泳法灵敏度高30倍。用同一原理的酶标记抗原融合电泳法监测L-氨基酸氧化酶的层析分离时,比酶活性法检测灵敏度高20倍。全部检测只需30分钟。初步观察商品眼镜蛇毒抗血清可对蛇毒15种成分产生免疫沉淀线,眼镜蛇的细胞毒与同科异科蛇毒的细胞毒素无交叉免疫反应。因此,采用本法可以确定不同蛇种蛇毒中所含的特异性抗原,获得蛇伤鉴别诊断的依据。     

THE NORMAL AND INDUCED OCCURRENCE OF CYANOPHYCIN INCLUSION BODIES IN SEVERAL BLUE-GREEN ALGAE1

Multi-L-arginyl-poly(L-aspartic acid) [arg-poly(asp)], the polypeptide component of the cyanophycin inclusion body, is found in cells of many blue-green algae. Formation of this material can be induced by a variety of treatments including the addition of excess nitrogen-containing compounds, the addition of specific inhibitors of macromolecular synthesis, and the exclusion of sulfur or phosphorus. Knowledge of the conditions that induce the synthesis of this polypeptide has made possible an ultrastructural survey to determine the presence and cellular location of cyanophycin bodies in a variety of cyanobacteria. Data presented show that certain strains of the unicellular genus Synechococcus Nägeli do not contain arg-poly(asp) under environmental conditions that markedly increase the level of such material in other cyanobacteria. In addition many strains show spatial localization of the cyanophycin bodies under normal growth conditions, and moreover the normal pattern is retained even when massive synthesis of arg-poly(asp) is induced. Finally there is no evidence that these inclusion bodies occur in certain beggiatoan gliding bacteria.     

USE OF ULTRASONIC SPECTROSCOPY AND VISCOSIMETRY FOR THE CHARACTERIZATION OF CHICKEN SKIN COLLAGEN IN COMPARISON WITH COLLAGENS FROM OTHER ANIMAL TISSUES

Use of traditional sources of collagen such as pork, bovine, and carp has some limitations. Chicken skin can be valuable alternative. In this work collagen was isolated from chicken skin using a modified procedure. Molecular properties of chicken collagen were analyzed and compared to collagen from other animal skins. Acid-soluble collagen type I was obtained with a yield of 25% and water content around 67%. Viscosimetry and ultrasonic spectroscopy were newly used for molecular characterization. By ultrasonic attenuation measurements, a pre-aggregation phase in the interval from 20°C to 27°C was observed, which is a proof of disaggregation and liquefaction. From 40°C upward, the liquefaction process finishes and aggregation continues. In a bovine sample this phenomenon starts at 40°C, in chicken at 50°C, and continues until 70°C. By viscosimetry, the denaturation temperature was confirmed as 40°C for bovine and 50°C for chicken collagen. Chicken collagen has a two times higher lysine level than bovine, which provides molecular stability side-chain interactions. With regard to higher thermal stability and favorable amino acid composition, waste chicken skin has the potential to be an excellent alternative source of raw collagen with applications in the food industry and biomedicine.     

渗透胁迫对小麦幼苗光合作用中氧和二氧化碳交换的影响

小麦（ＴｒｉｔｉｃｕｍａｅｓｔｉｖｕｍＬ．ｃｖ．１１６６）幼苗在不同渗透势的聚乙二醇溶液胁迫下,叶片的相对含水量和水势随渗透势下降递降,膜的相对透性增加,叶绿素和可溶性蛋白含量减少,核酮糖１,５－双磷酸浚化酶／加氧酶活性下降,而乙醇酸氧化酶活性则上升了。渗透胁迫对植物光合作用造成的不良影响,还表现为损害了光合放氧和二氧化碳的交换与代谢过程,其中二氧化碳受到的影响比氧敏感。     

HISTOPATHOLOGICAL AND HISTOCHEMICAL CHANGES IN HONEYBEE LARVAE (APIS MELLIFERA L.) AFTER INFECTION WITH BACILLUS LARVAE,THE CAUSATIVE AGENT OF AMERICAN FOULBROOD DISEASE

Morphological, histochemical and cytochemical changes were examined in honeybee larvae after infection with the bacterium Bacillus larvae. The results indicate cell necrosis in the midgut epithelium accompanied by increasing cell vacuolization and nuclear pyknosis following per os inoculation with B. larvae. Many autolysosomes were positive for acid phosphatase. Non-vacuolar acid phosphatase activity was also found in lysed cell compartments. No such activity was found in regenerative epithelial cells. Degradation of haemocytes, salivary glands and other tissues was also observed. Histochemical analyses after per cutaneous inoculation with B. larvae of three- and five-day-old honeybee larvae show intense non-vacuolar acid phosphatase activity followed by disintegration of infected salivary glands, epithelial cell cytoplasm and haemocytes.     

The role of xanthine oxidoreductase and uric acid in metabolic syndrome

Xanthine oxidoreductase (XOR) could contribute to the pathogenesis of metabolic syndrome through the oxidative stress and the inflammatory response induced by XOR-derived reactive oxygen species and uric acid. Hyperuricemia is strongly linked to hypertension, insulin resistance, obesity and hypertriglyceridemia. The serum level of XOR is correlated to triglyceride/high density lipoprotein cholesterol ratio, fasting glycemia, fasting insulinemia and insulin resistance index. Increased activity of endothelium-linked XOR may promote hypertension. In addition, XOR is implicated in pre-adipocyte differentiation and adipogenesis. XOR and uric acid play a role in cell transformation and proliferation as well as in the progression and metastatic process. Collected evidences confirm the contribution of XOR and uric acid in metabolic syndrome. However, in some circumstances XOR and uric acid may have anti-oxidant protective outcomes. The dual-face role of both XOR and uric acid explains the contradictory results obtained with XOR inhibitors and suggests caution in their therapeutic use.     

Oxidases and peroxidases in cardiovascular and lung disease: new concepts in reactive oxygen species signaling

Reactive oxygen species (ROS) are involved in numerous physiological and pathophysiological responses. Increasing evidence implicates ROS as signaling molecules involved in the propagation of cellular pathways. The NADPH oxidase (Nox) family of enzymes is a major source of ROS in the cell and has been related to the progression of many diseases and even environmental toxicity. The complexity of this family's effects on cellular processes stems from the fact that there are seven members, each with unique tissue distribution, cellular localization, and expression. Nox proteins also differ in activation mechanisms and the major ROS detected as their product. To add to this complexity, mounting evidence suggests that other cellular oxidases or their products may be involved in Nox regulation. The overall redox and metabolic status of the cell, specifically the mitochondria, also has implications on ROS signaling. Signaling of such molecules as electrophilic fatty acids has an impact on many redox-sensitive pathologies and thus, as anti-inflammatory molecules, contributes to the complexity of ROS regulation. This review is based on the proceedings of a recent international Oxidase Signaling Symposium at the University of Pittsburgh's Vascular Medicine Institute and Department of Pharmacology and Chemical Biology and encompasses further interaction and discussion among the presenters.