可口革囊星虫消化道的形态结构特征

采用解剖及组织学显微技术观察研究了可口革囊星虫消化道的形态及组织学结构特征。消化道始于口,经咽、食道、游离肠、螺旋肠(上行肠及下行肠)、直肠,止于肛门,各段从内向外均由粘膜层、粘膜下层、肌层及外膜组成。粘膜层表面具微绒毛及纤毛,微绒毛增加了消化道的表面积,纤毛的摆动有利于消化道内食物或消化后残渣的运送;螺旋肠及直肠的粘膜层中均有杯状细胞,在下行肠前段与中段及上行肠前段与直肠中尤其多,杯状细胞能够分泌粘液,保护粘膜上皮,润滑食物及残渣。粘膜下层为结缔组织,肌层为环肌,外膜为浆膜。肠沿纺锤肌盘绕的特点延长了食物的滞留、消化和吸收时间,弥补了缺乏消化腺的不足。肠各段除直肠肌层较厚外,其余各段肌层薄或不明显。     

可口革囊星虫体内纤溶酶的初步研究

目的从可口革囊星虫中寻找到纤溶酶,并对其进行初步研究。方法采用匀浆、抽提离心、Sephacryl S-300凝胶过滤等方法对可口革囊星虫纤溶酶初步分离,用纤维蛋白平板法和合成发色底物法检测其纤溶活性,并用该酶进行体外溶血凝块实验。结果可口革囊星虫内脏中存在纤溶酶。此酶既直接降解纤维蛋白又间接激活纤溶酶原,它对体外血凝块有明显溶解作用。结论可口革囊星虫内脏中存在着一种既具直接降解纤维蛋白作用又具激活纤溶酶原作用的纤溶酶。     

可口革囊星虫体腔中卵细胞大小组成的周年变化

在浙江温岭沿海逐月采样,显微观察了可口革囊星虫体腔中卵细胞大小组成的周年变化。3—5月,卵径30~90μm的卵细胞占大部分。6月,卵径70~130μm的卵细胞占多数。7月,大部分卵细胞卵径达100μm以上,其中130~150μm的卵细胞约占1/3。8月,卵细胞卵径均在110μm以上,其中130~150μm的卵细胞约占2/3。9月,仍多数卵细胞卵径在110μm以上,130~150μm的卵细胞约占1/3。10月,以110~140μm的卵细胞占多,40μm以下的卵细胞较少,大卵细胞开始退化。11月至翌年2月,随着大型卵细胞的逐渐退化,小型及中型卵细胞逐渐增多,2月仍可见少量未退化的大型卵细胞。分析认为,可口革囊星虫在浙江温岭的繁殖期主要在7—9月。     

可口革囊星虫消化道的形态及组织学结构

对可口革囊星虫(Phascolosoma esculenta)消化道各部分结构进行了形态学和组织学观察,表明可口革囊星虫消化道由翻吻、咽、食道、肠、直肠和肛门组成。翻吻收缩性很强;肠道极长,沿纺锤肌螺旋缠绕成肠索。消化道管壁由内向外分为粘膜层、粘膜下层、肌层和外膜。粘膜上皮主要由柱状细胞组成。除肠外,消化道上皮细胞均有发达的纤毛,肠上皮细胞主要具发达的微绒毛;粘膜下层及肌层的发达程度因消化道的部位不同而异。     

可口革囊星虫不同组织同工酶的比较

采用聚丙烯酰胺凝胶电泳(PAGE)技术,研究了可口革囊星虫5种组织(收吻肌、吻、肠、肾管及体壁)10种同工酶(ADH、GCDH、SDH、EST、LDH、ME、POD、SOD、MDH、CAT)的表达。结果表明:除CAT外,其余9种同工酶均得到了较为清晰的酶谱,除SOD和MDH在5种组织中表型相同外,其它7种同工酶的表达均具有明显的组织特异性。SDH-2、EST-1、LDH-1和POD-1为收吻肌所特有;ADH-3、ADH-4、GCDH-2、GCDH-4、GCDH-5、EST-10和LDH-6均为吻所特有;EST-4为肾管所特有。在肠中,ADH和GCDH均没有检测到活性。初步探讨了5种组织中各种同工酶的分布与活性及其与各组织生理功能的关系。     

可口革囊星虫的精子发生及精子结构

为了探究可口革囊星虫(Phascolosoma esculenta)精子发生过程及结构上的特殊性,用显微及亚显微技术研究了可口革囊星虫的精子发生和精子结构。可口革囊星虫的精巢位于收吻肌基部,为一曲折的带状组织。成熟精巢内可观察到精原细胞、精母细胞以及精细胞等各阶段的生精细胞。在精子形成早期,很多精细胞脱离精巢,以精细胞团的形式掉落到体腔中。精细胞团内的精细胞同步发育为精子后,脱离精子团进入肾管。成熟精子由头部和尾部组成。头部由钟形顶体与鼓形细胞核构成。顶体后段下包于精核的前端。顶体分内、中、外三层,外层有横隔;顶体下腔内有颗粒状物质不均匀分布,中央有一束丝状纤维组成的顶体棒。核物质电子密度高,核内含空泡。无核前窝,具浅的核后窝。尾部分中段和末段,中段由6个(偶见5个或7个)线粒体围绕近、远端中心粒构成;末段细长鞭状,由轴丝及包绕轴丝的质膜组成,轴丝为典型的"9 2"结构。分析认为:可口革囊星虫精子发生过程以及超微结构上存在特殊的结构与机制:①精细胞团保证了精子形成的同步性;②顶体后段下包于精核的前端使精子头部小而灵巧,利于快速运动;③顶体的横隔使精子顶体的牢固性增强,确保受精时顶体反应的正常进行;④中段较多的线粒体使精子具有更强的环境适应性,有利于有效的受精。     

可口革囊星虫胚胎与幼体的发育

解剖经降温8-10℃刺激30-40h后的可口革囊星虫(Phascolosomaesculenta)亲体取得精、卵,用人工授精法获得的受精卵进行胚胎和幼体发育的研究。结果表明:可口革囊星虫的卵为圆形或椭圆形,卵的大小为120μm×142μm-125μm×148μm。精子由头部、中段和尾部组成,全长约为32-40μm。在海水盐度23.8ppt、pH8.1、温度15℃的条件下,精、卵不能受精;在海水盐度23.8ppt、pH8.1、温度20℃的条件下,精卵能受精,但胚胎发育到原肠胚期停止;在海水盐度23.8ppt、pH8.1条件下,25℃、30℃和35℃的不同水温,精卵均能受精,受精卵发育到初期海球幼虫分别需要72h0min、47h08min和42h53min。在海水盐度23.8-25.5ppt,pH8.1-8.2,温度28-30℃条件下,从初期海球幼虫发育到稚虫需要15-20d。     

可口革囊星虫受精过程及早期卵裂的细胞学变化

为探究可口革囊星虫(Phascolosoma esculenta)受精过程中精子入卵、极体排放、雌雄原核的形成与结合以及早期卵裂的特点,为胚胎发育机制研究奠定基础及指导人工育苗,显微观察了可口革囊星虫卵母细胞的形态;用荧光染色剂HOECHST33258对已固定的成熟卵及受精卵进行染色的方法,在荧光显微镜下观察了可口革囊星虫受精过程及第一与第二次卵裂的细胞学变化。成熟卵呈椭圆形,卵膜较厚,核区偏位,染色体排列整齐。在水温30℃-31℃、盐度23条件下,授精后5min-10min,精子完成入卵;授精后10min-20min,完成第一次减数分裂、释放第一极体;授精后20min-30min,完成第二次减数分裂、放出第二极体,部分受精卵的第一极体分裂为二;授精后30min-40min,雌、雄原核形成,并相互靠近向卵中央迁移;授精后40min-50min,雌、雄原核在卵中央结合成合子核;授精后50min-70min及70min-90min分别完成第一及第二次卵裂。可口革囊星虫的受精过程及早期卵裂的细胞学特点为:1)成熟卵是处在第一次减数分裂中期的初级卵母细胞,具有受精能力;2)精子入卵位点是随机的,存在多精入卵现象;3)雌、雄原核以融合方式结合成合子核;4)第一及第二次卵裂均为经裂、不等裂。     

可口革囊星虫肾管肌组织的结构特征

采用显微及亚显微技术观察了可1：7革囊星虫肾管肌组织的结构特征。肾管肌组织位于柱状上皮层下,由纵肌及环肌组成。肌细胞（肌纤维）呈长梭形,核位于细胞边缘并明显突向细胞外基质中,核周围有较多线粒体及少量内质网。肌纤维表面有许多囊状或指状突起的肌质囊,内含肌浆、光面内质网、线粒体及糖原颗粒。肌质囊之间的肌膜内面具膜相关电子致密斑。肌纤维内含粗、细两种肌丝,细肌丝围绕在粗肌丝周围,在肌丝之间分布有糖原颗粒、线粒体及胞质致密体。线粒体及糖原为肌纤维的代谢提供能量,肌组织的收缩对促进肾管的过滤排泄及繁殖时配子进入肾管可能起重要作用。     

可口革囊星虫体液细胞的显微和亚显微结构

本文利用光学显微镜和透射电镜研究了可口革囊星虫体液细胞的显微和亚显微结构。研究发现该星虫体液中含有红细胞、颗粒细胞、桑椹胚样细胞和卵母细胞。可口革囊星虫的红细胞数量较多,红细胞呈盘状,从一边或两边略凹陷,最典型的是从三边发育的二裂片内陷,直径为24μm-32μm。圆形红细胞,核位于中央,细胞内很少有细胞器,细胞质内充满嗜锇的血红蛋白。体腔液内含有两种类型的颗粒细胞。Ⅰ型细胞含有大量的小颗粒,是典型的变形细胞,细胞质里含有大量的线粒体、最典型的高尔基体、内质网和各种形状不一的小池。Ⅱ型细胞含有大量的嗜锇细胞质。在可口革囊星虫体液中含有不同发育时期的卵母细胞。     

HPLC preparation of fish waste hydrolysate fractions. Effect on guinea pig ileum and ACE activity

The effect of RP-HPLC-purified fractions of fish waste hydrolysates issued from three fish industries was tested on guinea pig ileum in order to examine the presence of opioid molecules. The evaluation of anti-hypertensive activities of whole hydrolysates and fractions were also tested, monitoring the ability of the fraction to inhibit the activity of angiotensin I-converting enzyme involved in hypertension regulation. Sardine autolysate and cod head hydrolysate powder (50 microg) were able to inhibit near 30% of ACE activity, whereas 50 microg of shrimp hydrolysate allows the inhibition of 57% of ACE activity. HPLC fractionation of cod head hydrolysate and sardine autolysate was necessary to evidence biological activity, whereas HPLC separation of shrimp hydrolysate exhibited low biological activity fractions. Further studies are necessary to characterise bioactive molecules from cod head alcalase hydrolysate and from sardine autolysate.     

利用制霉菌素浓缩处理高频率地检出黑曲霉营养缺陷型突变株

以亚硝酸诱变处理黑曲霉Ｎ３４３和黑曲霉ＵＶ－１１的分生孢子,然后将它们培养在完全培养基平板上至第二代分生孢子长出,收集这些孢子以制霉菌素浓缩处理,结果以３．８～０．３％的高频率得到１９株稳定的营养缺陷型突变株：从黑曲霉Ｎ３４３获得３株维生素类的缺陷型,它们分别为双缺或叁缺;从黑曲霉ＵＶ－１１获得１６株特殊的脯氨酸营养缺陷型,进一步分析表明这是由于从谷氨酸合成脯氨酸或精氨酸的代谢阻断所致。所有以上突变株形态与各自原养型相似,但在产酶活力方面均为负突变     

Improvement of ACE inhibitory activity of chitooligosaccharides (COS) by carboxyl modification

In the present research, chitooligosaccharides (COS) were carboxylated with -COCH(2)CH(2)COO(-) groups to obtain specific structural features similar to Captopril. Angiotensin I converting enzyme (ACE) inhibitory activity of carboxylated COS was studied and observed to enhance its activity with increased substitution degree. Further, Lineweaver-Burk plot analysis revealed that inhibition was competitive via obligatory binding site of the enzyme. This was accompanied with substitution of positively charged quarternized amino groups to COS with different substitution degrees, in which negative impact on ACE inhibition was observed.     

Inhibition of angiotensin converting enzyme (ACE) activity by flavan-3-ols and procyanidins

It was determined that flavan-3-ols and procyanidins have an inhibitory effect on angiotensin I converting enzyme (ACE) activity, and the effect was dependent on the number of epicatechin units forming the procyanidin. The inhibition by flavan-3-ols and procyanidins was competitive with the two substrates assayed: N-hippuryl-L-histidyl-L-leucine (HHL) and N-[3-(2-furyl)acryloyl]-L-phenylalanylglycylglycine (FAPGG). Tetramer and hexamer fractions were the more potent inhibitors, showing Ki of 5.6 and 4.7 microM, respectively. As ACE is a membrane protein, the interaction of flavanols and procyanidins with the enzyme could be related to the number of hydroxyl groups on the procyanidins, which determine their capacity to be adsorbed on the membrane surface.     

Determining the phenolic components by using HPLC and biological activity of Centaurea triumfetti

Abstract

Centaurea species belonging to Asteraceae family are traditionally used herbs in treating many diseases in Anatolia. In the present study, I aimed to determine the biological activities and phenolic contents of the extracts of the aerial parts of Centaurea triumfetti collected from areas with extreme climatic conditions. The determination of antioxidant activity was performed using in vitro assays such as ABTS (2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) and DPPH (1,1-diphenyl-2-picrylhydrazyl). Disc diffusion and microdilution methods were used for determine of antimicrobial activity. Besides that, the diabetes-related inhibition activities of enzymes such as alpha-glucosidase and alpha-amylase were determined using colorimetric methods. The phenolic components of the extracts were analysed by using HPLC. The biological activity of ethyl acetate extracts of the stem was found to be higher those than that of other extracts. This extract was also found to have high antimicrobial and enzyme inhibition activity. Moreover, the major component of the extract was found to be chlorogenic acid. In conclusion, these results suggest that C. triumfetti has the potential for pharmaceutical usage.