Purification and characterization of the carbonic anhydrase enzyme from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on enzyme activity

In this study, the carbonic anhydrase (CA) enzyme was purified from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney with a specific activity of 603.77 EU/mg and a yield of 35.5% using Sepharose-4B-l-tyrosine- sulphanilamide affinity column chromatography. For determining the enzyme purity and subunit molecular mass, sodiumdodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) was performed and single band was observed. The molecular mass of subunit was found approximately 29.71 kDa. The optimum temperature, activation energy (E_a), activation enthalpy (ΔH) and Q₁₀ values were obtained from Arrhenius plot. K_m and V_max values for p-nitrophenyl acetate of the purified enzyme were calculated from Lineweaver-Burk graphs. In addition, the inhibitory effects of different heavy metal ions (Fe²⁺, Pb²⁺, Co²⁺, Ag⁺ and Cu²⁺) on Black Sea trout kidney tissue CA enzyme activities were investigated by using esterase method under in vitro conditions. The heavy metal concentrations inhibiting 50% of enzyme activity (IC₅₀) were obtained. Finally K_i values and inhibition types were calculated from Lineweaver-Burk graphs.     

Development of an IgG-Fc fusion COVID-19 subunit vaccine,AKS-452

AKS-452 is a biologically-engineered vaccine comprising an Fc fusion protein of the SARS-CoV-2 viral spike protein receptor binding domain antigen (Ag) and human IgG1 Fc (SP/RBD-Fc) in clinical development for the induction and augmentation of neutralizing IgG titers against SARS-CoV-2 viral infection to address the COVID-19 pandemic. The Fc moiety is designed to enhance immunogenicity by increasing uptake via Fc-receptors (FcγR) on Ag-presenting cells (APCs) and prolonging exposure due to neonatal Fc receptor (FcRn) recycling. AKS-452 induced approximately 20-fold greater neutralizing IgG titers in mice relative to those induced by SP/RBD without the Fc moiety and induced comparable long-term neutralizing titers with a single dose vs. two doses. To further enhance immunogenicity, AKS-452 was evaluated in formulations containing a panel of adjuvants in which the water-in-oil adjuvant, Montanide™ ISA 720, enhanced neutralizing IgG titers by approximately 7-fold after one and two doses in mice, including the neutralization of live SARS-CoV-2 virus infection of VERO-E6 cells. Furthermore, ISA 720-adjuvanted AKS-452 was immunogenic in rabbits and non-human primates (NHPs) and protected from infection and clinical symptoms with live SARS-CoV-2 virus in NHPs (USA-WA1/2020 viral strain) and the K18 human ACE2-trangenic (K18-huACE2-Tg) mouse (South African B.1.351 viral variant). These preclinical studies support the initiation of Phase I clinical studies with adjuvanted AKS-452 with the expectation that this room-temperature stable, Fc-fusion subunit vaccine can be rapidly and inexpensively manufactured to provide billions of doses per year especially in regions where the cold-chain is difficult to maintain.     

Extracellular Enzyme Activities by Basidiobolus and Conidiobolus Isolates on Solid Media./Extrazelluläre Enzymaktivitäten bei Basidiobolus- und Condiobolus-Isólaten auf festen Medien

Summary: Fifty-seven isolates of Basidiobolus from reptiles and amphibians, and 7 other obtained from the American Type Culture Collection (ATCC) in Maryland were studied for their extracellular enzyme activities on solid media. The Conidiobolus isolates studied included 4 recovered from Nigerian soil and additional 4 obtained from the ATCC All these isolates produced active extracellular lipase and protease and failed to exhibit amylase, deoxyribonuclease and ribonuclease activities. The significance of the findings is discussed.
Zusammenfassung: Fünfundsiebzig Basidiobolus-Isolate aus Reptilien und Amphibien und sieben weitere aus der American Type Culture Collection (ATCC) in Maryland wurden auf Aktivtäten extrazellulärer Enzyme auf festen Medien untersucht Die untersuchten Conidiobolus-Isolate schlossen vier aus nigerianischen Böden und vier weitere aus der ATCC ein. Alle Isolate zeigten extrazellulär Lipase- und Proteaseaktivität Amylase-, Desoxyribonuclease- und Ribonucleaseaktivität war jedoch nicht nachzuweisen. Die Bedeutung dieser Befunde wird diskutiert.     

日本血吸虫酚氧化酶的组化定位研究

目的观察酚氧化酶(phenol oxidase,PO)在日本血吸虫成虫的组织学分布.方法酚氧化酶组化方法:将42 d虫龄的日本血吸虫活成虫置含25 mmol/L邻苯二酚的磷酸盐缓冲液(pH 6.8)中,37℃孵育30 min后,转移虫体至载玻片上,吸去虫体表面液体,将其中一部分虫体置Olym-pus显微镜下观察酚氧化酶在虫体的组织学分布.荧光组织化学方法:经上述过程处理后,在另一部分虫体上滴加2滴含0.05%戊巴比妥钠的PBS溶液,然后置Leica荧光显微镜下观察酚氧化酶在虫体的组织学分布.结果酶组织化学方法显示,日本血吸虫酚氧化酶仅分布于雌虫卵黄腺及子宫内虫卵卵壳表面,呈现棕褐色显色反应;雌虫卵巢和雄虫均未发现酚氧化酶活性.然而,荧光组织化学方法显示,酚氧化酶除主要分布于雌虫卵黄腺及子宫内虫卵卵壳表面,呈现强荧光外,还少量分布于雄虫体壁表层,呈现弱荧光反应.结论不仅日本血吸虫雌虫含有酚氧化酶活性,而且雄虫也含酚氧化酶活性,只不过其含量少、活性低.荧光组织化学方法能更灵敏地显示日本血吸虫酚氧化酶活性,更适用于日本血吸虫酚氧化酶的组织学定位.     

110例嗜酸细胞增多性非变应性鼻炎的液氮冷冻治疗

１１０例嗜酸细胞增多性非变应性鼻炎的液氮冷冻治疗山西医学院第二附属医院（０３０００１）牛玉梅，刘学仁按Ｍｇｇｉｎｄ将常年性鼻炎分为三类：一类常年性变态反应性鼻炎；二类非变态反应性鼻炎伴鼻分泌物嗜酸细胞增多综合征也叫嗜酸细胞增多性非变态反应性鼻炎（Ｅｏｓｉｎｏｐｈｉｌｉｃｎｏｎａｌｌｅｒａｉｃｒｈｉｎｉｔｓ，ＥＮＲ）；三类自主神经性常年性鼻炎［１］。我们将ＥＮＲ中找不到致敏物，而与冷热空气有关与情绪无关的１１０例患者进行了液氮冷冻治疗。临床资料８０２例常年性鼻炎患者进行皮肤激发试验。４３７例找出了致敏物，进行特异性过敏诱因脱敏治疗或抗过敏药物治疗。３６５例试验阳性，其中有１３５例与冷热空气有关。ｌｌ０例行冷冻治疗。１ｌ０例中男５０例，女６０例；年龄最大６５岁，最小１６岁。１１０例均有间歇性的连续喷嚏发作，浆液性或粘液性鼻分泌物增多和鼻粘膜非充血性肿胀引起的堵塞，无因吸入致敏原诱发症状的病史；血清ＩｇＥ值不升高，特异性皮肤试验结果为阳性；鼻分泌物中嗜酸细胞阳性。方法：先将鼻腔内喷入１％地卡因溶液３次粘膜表面麻醉，用卷好的４厘米长、０．４～０．６厘米粗的棉棒沾上液氮在鼻镜直视下迅速插入鼻腔内。时间约１分钟左右（冷     

自身免疫性白内障的晶状体代谢与生理改变研究

目的 :探讨自身免疫性白内障 (AIC)的病理生理学机制。　方法 :采用酶组织化学方法以及晶状体生理功能检查 ,观察AIC模型动物晶状体生理功能与组织内主要酶活性的变化。　结果 :晶状体组织内Na K ATP酶活性降低与晶状体调节力下降、裂隙灯下显微镜混浊改变之间存在着相关性。　结论 :自身免疫性晶状体损伤 ,进而造成晶状体组织内相关酶代谢异常是AIC的病理基础     

Head-impulse testing in Fabry disease - vestibular function in male and female patients

Aim : To study the prevalence of peripheral vestibular deficit in male and female patients with Fabry disease and to assess the effect of enzyme replacement therapy (ERT) on peripheral vestibular function using quantitative head-impulse testing. Methods : Using dual search-coils the vestibulo-ocular reflex during rapid rotational head thrusts to both sides was recorded in 21 patients (13 male, 8 female) with Fabry disease prior to ERT initiation. ERT consisted of infusions of gene-activated human α-galactosidase A (agalsidase alfa; Replagal™) every 2 weeks at doses of 0.2mg/kg. Eight patients were tested again approximately 6 and 12 months after the initiation of ERT. Results : At baseline examination, 15 of the patients with Fabry disease (71%; 11 males, 4 females) showed reduced peripheral vestibular function. The deficit was unilateral in nine patients (3 females) and bilateral in six patients (1 female). The severity of the vestibular deficit was not significantly different between male and female patients. After 12 months of ERT, the average vestibular deficit on the weaker side tended to improve; however, the change was not significant ( p = 0.10).
Conclusion : Fabry disease affects peripheral vestibular function in both male and female patients. Females seem to be affected less frequently than males, but, on average, vestibular deficits are not different between the two groups. To confirm or reject the tendency for vestibular improvement during ERT, more patients need to be tested and longer follow-up periods are required.