Polyethylene glycol fusion repair of severed rat sciatic nerves reestablishes axonal continuity and reorganizes sensory terminal fields in the spinal cord

Peripheral nerve injuries result in the rapid degeneration of distal nerve segments and immediate loss of motor and sensory functions;behavioral recovery is typically poor.We used a plasmalemmal fusogen,polyethylene glycol(PEG),to immediately fuse closely apposed open ends of severed proximal and distal axons in rat sciatic nerves.We have previously reported that sciatic nerve axons repaired by PEG-fusion do not undergo Wallerian degeneration,and PEG-fused animals exhibit rapid(within 2–6 weeks)and extensive locomotor recovery.Furthermore,our previous report showed that PEG-fusion of severed sciatic motor axons was non-specific,i.e.,spinal motoneurons in PEG-fused animals were found to project to appropriate as well as inappropriate target muscles.In this study,we examined the consequences of PEG-fusion for sensory axons of the sciatic nerve.Young adult male and female rats(Sprague–Dawley)received either a unilateral single cut or ablation injury to the sciatic nerve and subsequent repair with or without(Negative Control)the application of PEG.Compound action potentials recorded immediately after PEG-fusion repair confirmed conduction across the injury site.The success of PEG-fusion was confirmed through Sciatic Functional Index testing with PEG-fused animals showing improvement in locomotor function beginning at 35 days postoperatively.At 2–42 days postoperatively,we anterogradely labeled sensory afferents from the dorsal aspect of the hindpaw following bilateral intradermal injection of wheat germ agglutinin conjugated horseradish peroxidase.PEG-fusion repair reestablished axonal continuity.Compared to unoperated animals,labeled sensory afferents ipsilateral to the injury in PEG-fused animals were found in the appropriate area of the dorsal horn,as well as inappropriate mediolateral and rostrocaudal areas.Unexpectedly,despite having intact peripheral nerves,similar reorganizations of labeled sensory afferents were also observed contralateral to the injury and repair.This central reorganization may contribute to the improved behavioral recovery seen after PEG-fusion repair,supporting the use of this novel repair methodology over currently available treatments.

Biological Characteristics of Serum Agglutinin from Portunus trituberculatus

The agglutination characteristics of serum agglutinin from Portunus trituberculatus were studied in this paper. The results showed that the serum agglutinin had no agglutination to crucian carp, Chinese soft shell turtle, grass carp, chicken or human group A, B or O blood cells, but had strong agglutination to blood cells of mice and rabbits. The activities of serum agglutinin from Portunus trituberculatus to mice blood cells reached 210, and to rabbit blood cells reached 28. Salinity had a greater effect on agglutinin activity. When the Na Cl concentration exceeded 0.6 mol/L,the serum agglutinin from Portunus trituberculatus was basically inactivated. The optimum p H for agglutinin activity was 6.0-7.4. The serum agglutinin from Portunus trituberculatus had obvious dependence to Ca2 ＋and Mg2 ＋, and EDTA could significantly inhibit its activity. The results of sugar inhibition test showed that the activity of agglutinin from Portunus trituberculatus can be specifically inhibited by N-acetylglucosamine and N-acetylmannosamine. The serum agglutinin from Portunus trituberculatus was isolated by ammonium sulfate gradient precipitation, and its activity was highest by the 25% ammonium sulfate precipitation system. The SDS polyacrylamide gel electrophoresis（SDS-PAGE） showed that the protein bands were mainly distributed within 72-95 ku.

Anti-nutritional characteristics and mechanism of soybean agglutinin

Soybean agglutinin(SBA)is an important anti-nutritional factor in soybean.SBA can induce animal growth inhibition,cause pathological changes of intestinal tissue,and decrease in the immune system functioning.Recently,a great deal of research has been done on the effects of SBA on cell morphology,division,apoptosis,autophagy,as well as the correlated signal transduction pathway.This review mainly covers the chemical and biological characteristics of SBA,describes the multifaceted aspects of SBA anti-nutritional functions,and highlights the possible cellular and molecular mechanism of anti-nutritional effects of SBA.This review has important implications for the prevention and treatment of SBA-induced diseases,drug development,processing techniques of plant products,prevention of food-borne toxins,as well as human and animal health protection.

Exploring the Insecticidal Potentiality of <i>Amorphophallus paeonifolius</i>Tuber Agglutinin in Hemipteran Pest Management

Hemipteran group of sap sucking insect pests cause worldwide crop destruction. The role of mannose specific monocot lectins have recently been worked out in hemipteran pest management. The present article demonstrates the insecticidal efficacy of a new mannose specific agglutinin, isolated from tubers of Amorphophallus paeonifolius (AMTL) against a wide range of hemipteran insects. The 25 kDa dimeric protein was found to inhibit the survivability of hemipteran insects namely, Lipaphis erysimi, Aphis gossypii and Dysdercus cingulatus quite efficiently, as analysed by synthetic diet based bioassay experiments. Surface Plasmon Resonance study detected binding of insecticidal AMTL to insect gut brush border membrane vesicle (BBMV) protein, an absolute prerequisite for conferring toxicity against target insects. Further ligand blot analysis spotted a ~74 kDa glycoprotein as putative receptor of AMTL from the total BBMV protein fraction of Lipaphis erysimi. Phylogenetic analysis showed a significant relatedness of AMTL to the previously established monocot lectin Galanthus nivalis agglutinin (GNA) in terms of their conserved mannose binding domains, agglutinating ability of rabbit erythrocytes and insecticidal efficacies. These information project AMTL as a promising candidate in preventing crop loss caused due to hemipteran insect attack.

Agglutinin activity in hemolymph of Chinese shrimp and induction effect of Vibrio

Experiments showed the agglutinin activity in the hemolymph of both spring wild parents and autumncultured individuals of the shrimp Penaeus chinensis Osbeck with the body length of 12 - 16 cm as all 32. Thehemolymph of the above shrimps showed agglutinant reaction to all A-, B- and O-type human erythrocytes, with thereaction titer equal to that of rabbit blood cells. However, the agglutinin activity level was enhanced after being in-duced by Vibro, which was 128 after 24 h of induction and reached the peak value 256 after 48 h of induction, thendecreased gradually.

SCREENING OF AGGLUTININS IN MARINE ALGAE FROM FUJIAN COAST OF CHINA

Thirty three species of marine algae belonging to Rhodophyta, Phaeophyta and Chlorophyta from the Fujian coast were examined for agglutinins with different animal and human erythrocytes. Protein extracts from 26 species were active against at least one type of the erythrocytes tested. There were 3 species ( Grateloupia imbricata, Ishige foliacea and Entermorpha prolifera ) whose extracts could agglutinate all the erythrocytes used. The lowest protein concentration required to produce erythrocyte agglutination varied remarkably, from 3.1 μg/ml to 500 μg/ml . The strongest activity was found in the agglutination of rabbit erythrocytes by Gloiopeltis furcata extract. Inhibition assays performed with nine mono and bisaccharides indicated that agglutinations of rabbit erythrocytes by extracts of 7 species were inhibited by one or more types of the sugars assayed. The agglutinating activity shown by extracts of most species was not affected when the test solution was heated to 90℃, but was lost at 95℃-100℃. A few extracts lost their activity at 60℃, 65℃ and 75 ℃, respectively.

Application of an Optical Biosensor to Study the Interaction between Porphyrins and Wheat Germ Agglutinin

An optical biosensor with a stirred cuvette has been used to monitor the interaction of immobilized wheat germ agglutinin （WGA） with two water-soluble cationic porphyrins, The association constants （Ka） of the free base porphyrin and its Zn（Ⅱ） complex form were 2. 66 and 27.31 × 10^5 l,/mol at 20 ℃ respectively. The interactions of the free base porphyrin were further investigated at temperatures between 15 ℃ and 37 ℃, The thermodynamics parameters, changes in free energy, enthalpy and entropy, were -31.23, 22.92, 54.15 ld/mol respectively. The heat capacity change was -355.53 J · mol^-1·K^-1 The binding was driven by entropic contribution, and showed strong enthalpy-entropy compensation. It was governed primarily by hydrophobic forces.

Cold agglutinin disease associated with mycoplasma infection in an individual with type 2 diabetes: An atypical case

Cold Agglutinin Disease is a hemolytic anemia associated with cold reactive autoantibodies. Although the acute form of cold agglutinin disease can be attributed to autoimmune or infectious diseases and lymphoproliferative diseases, it has, to the best of our knowledge, so far,never been reported as secondary to mycoplasma pneumonia in a type 2 diabetic individual. In this paper, we report a case of cold agglutinin disease following mycoplasma pneumonia in a 47-year-old female patient with type 2 diabetes and schizophrenia. Cold agglutinin test and direct Coombs’ test was positive. Anti-mycoplasma anti-body titre by complement fixation was high (Anti-IgG was negative and Anti-C3d was positive) and was accompanied by hemolytic anemia. Her general condition, including Cold Agglutinin Disease improved after conservative therapy with antibiotics, hypoglycemic agents and short-term use of steroids. The patient recuperated and was discharged in good health after 7 days’ stay in the hospital. She remains clinically well with no recurrence of anemia. The simultaneous occurrence of cold agglutinin disease, mycoplasma infection and diabetes mellitus is rare and accumulation of case reports is required to gain better insight of this case scenario.

Improved Recovery of Erythropoietin and Darbepoetin from Equine Plasma by the Application of a Wheat Germ Agglutinin Mediated Pre-Extraction Prior to Immunoaffinity Chromatography

We describe a two-step method that uses wheat germ agglutinin immobilized on Sepharose gel followed by immunoaffinity chromatography (IAC) to extract recombinant human erythropoietin and Darbepoetin from equine plasma. Lectin affinity chromatography was shown to be an effective approach for isolating these epoetins from plasma and in combination with IAC;this method gave superior recovery when compared to the use of the latter technique alone. Moreover, due to the ease with which it can be scaled up, it is particularly well suited for pre-concentrating larger volumes of samples prior to IAC and this provides a facile way of improving the overall sensitivity with which these foreign proteins can be detected in equine plasma.

Insight to the Mode of Action of <i>Allium sativum</i>Leaf Agglutinin (ASAL) Expressing in T₃Rice Lines on Brown Planthopper

Brown planthopper, the sap sucking hemipteran pest, is one of the major contributors to the yield loss of rice through the world. To combat the situation researchers are interested identifying genes from plant origin having potentiality to develop hemipteran pest resistance. Interestingly, it was observed that rice plants expressing ASAL, a monocot mannose binding lectin, showed significant resistance to brown planthopper and green leafhopper. Additionally, antibiotic resistant marker gene free ASAL expressing rice lines were developed to overcome the biosafety issues. However, the basis behind the resistance against planthoppers is still not clearly understood. Ligand blot assay was performed with total BBMV protein from BPH and a ~56 kDa receptor protein was detected. LC MS/MS analysis revealed that the receptor protein is NADH quinone oxidoreductase (NQO), a key player in electron transport chain, insect defense response and male/female gametogenesis. Presumably interaction of ASAL with NQO may lead to toxicity and loss of fecundity among BPH feeding on ASAL expressing transgenic rice plants. These findings provide a stable scientific basis for considering these transgenic ASAL expressing rice plants as significant product for combating BPH attack associated yield loss of rice.

Comparative Studies of the Carbohydrate of Human Gamma-Crystallins from Fetal and Adult Lenses with Agglutinins

Using gel chromatography of Sephadex G-75 superfine connectedwith Sephadex G-50 fine column,three human γ- crystallins(γ1,γ2,γ3)couldbe obtained.Seven agglutinins(LCA,SBA,DBA,PNA,BSL,RCA and UEA)were used to detect the sugar of sub-γ-crystallins,which had been transferredto nitrocellulose membrane and finally stained with ABC reagents and the sub-strate of HPR.These results suggested that γ2-and γ3-crystallin contain sugar,but γ1-crystallin has no sugar.There is a decrease of carbohydrate of γ2 and γ3as...

THE APPLICATION OF ION EXCHANGERS IN THE PREPARTION OF WHEAT AGGLUTININ

In order to refine the products of wheat germ agglutinin(WGA),several ion exchangers,such as D261,732,DEAE-cellulose-32 and CM-cellulose-32, have been used to removed proteins and pigments with different colors,such as brown,red,yellow,green and black,in the extracts of wheat germ.The WGA obtained from this procedure has higher hemagglutination activity than that available from sigma Co. The minimum hemagglutination dose of the purified WGA for an equal volume of a 2% type A red blood cells is 4μg/ml.WGA is a mixture of isolectins with different isoelectric points,5.9,6.2 and 6.8.Their molecular weight identified by SDS-polyacrylamide gel electrophoresis are 15,000 dalton,18,000 dalton and 35,000 dalton,respectively.

Reduction of High-Titer Cold Agglutinins by Plasmapheresis to Resolve Serological Problem in a Patient

Background: Cold agglutinins are auto-antibodies that can be a nuisance in cross matching and in blood grouping. Here we report an unusual case of a high titer and wide amplitude cold agglutinin reduced by plasmapheresis. Methods and Materials: A 56-year-old man with severe anemia requested a transfusion of red blood cells. However, there was a problem in blood for blood grouping. The discrepancy of blood typing was subsequently resolved using group O absorbed plasma along with repetition of forward grouping with warm-washed red blood cells. The presence of high-thermal-amplitude and a high-titer anti-I cold agglutinin were detected in further serologic investigation. It revealed reactivity against autologous and adult O red blood cells at 37°C by the thermal amplitude screening test, and demonstrated a very high titer of 65,536 against adult O cells by titration studies at 4°C. The patient received two plasma exchange sessions of 1.5 plasma volumes each. There was a significant reduction of the titer of cold agglutinins and of the thermal amplitude by plasmapheresis as well (p < 0.01). Results: After successful cross-matching with post plasma exchanges, four units of red blood cells were infused to the patient without any hemolysis symptoms or signs. Conclusions: We now reported a patient with abnormally ascended titer of cold agglutinins and wide-thermal-amplitude, but we also successfully performed ABO typing and cross matching after 2 plasma exchange sessions of 1.5 plasma volumes each.

Cold Agglutinin Disease Prevalence and Immunoglobulin M levels in Patients with Splenomegaly from Endemic Areas for Malaria in Rwanda

Background: Inhabitants from malaria endemic zones often present with enlarged spleen, mainly due to hyper reactive malarial splenomegaly (HMS), and it is seen more commonly associated with elevated levels of Immunoglobulin M (IgM). Cold agglutinin disease is an acquired autoimmune hemolytic anemia (AIHA) that is usually due to cold-reacting IgM autoantibodies directed against red cell antigens. The study was conducted in response to the observed high frequency of transfusion dependent anemia in patients with splenomegaly from malaria endemic zones in southern province of Rwanda. The objectives of this study were to determine the prevalence of cold agglutinin disease and to assess the distribution of IgM antibodies among these patients. Methodology: This was a descriptive, cross-sectional study conducted over a period of six months from June 2016 to December 2016. The study enrolled adult population from malaria endemic areas of the southern province of Rwanda with unexplained splenomegaly. Blood samples for testing IgM levels, cold agglutinin, FBC, and markers of hemolysis were collected from peripheral health settings and analyzed at the laboratory of Butare University Teaching Hospital. Results: During the study period, we enrolled 188 participants with enlarged spleen. One hundred twenty-five (66%) were females and 34% were males. The mean (±SD) age of the study participants was 35.6 ± 15.2 years. Out of 188 participants, only 4.8% were found with significantly positive Cold Agglutinin Titer (CAT) (≥1:64) and 84% of participants were found to have elevated serum IgM level. Hemoglobin level, white blood count and platelets count decrease with severity of splenomegaly, while IgM level increases with spleen size. Conclusion: Among patients with splenomegaly from malaria endemic areas, IgM levels correlate with the stages of splenomegaly while cold agglutinin disease plays a small role in the etiology of anemia.

酵母细胞表面展示技术

酵母细胞表达体系具备较为完善的蛋白质翻译后修饰和分泌的机制。以酵母为基础的细胞表面展示技术是一项新兴的真核蛋白展示技术,已成功应用于蛋白质识别、蛋白质的固定化和定向进化研究,成为了蛋白质工程研究的重要工具。根据与酵母细胞壁的外源目的蛋白融合部位的不同,酿酒酵母表面展示系统主要分为凝集素系统和絮凝素系统2大系统。将该技术引入动物营养研究领域,有望通过诱导宿主产生原虫或甲烷菌的抗体,对瘤胃微生态进行调控,从而以减少甲烷排放;生产高活性的全细胞催化剂,以提高反刍动物和单胃动物对纤维物质的利用效率。为此,本文就该技术的原理、特点和应用领域进行了综述。

自组装嵌合纤维小体酿酒酵母菌群的乙醇发酵研究

在可共代谢葡萄糖木糖的二倍体酿酒酵母Y6细胞表面,通过组成型强启动子（PGK）控制的a凝集素表面展示系统,构建一个可共代谢纤维素半纤维素的双支架纤维小体。通过免疫荧光、流式细胞术及酶催化活性的测定,证明纤维小体结构能正确组装并以活性形式固定在细胞表面。以纤维素和半纤维素为底物进行乙醇发酵实验,60 h时最大乙醇产量达1.98 g/L,乙醇产率为0.33 g/g,相当于理论值的65%。结果证明,表面展示嵌合纤维小体的酿酒酵母可实现共降解纤维素和半纤维素生产乙醇的目标。

蓝桉果免疫调节作用的实验研究

目的探讨蓝按果对小鼠巨噬细胞吞噬功能、免疲器官厦抗体生成的影响．方法采用腹腔巨噬细胞吞噬测定方法、免疫器官重量法及凝集素测定方法，观察吞噬百分率、吞噬指教、胸腺指教和脾脏指数、血清凝集素的生成．结果蓝按果溶液具有增强小鼠腹腔巨噬细胞吞噬功能的作用，能明显增加小鼠免疫器官的重量和明显提高小鼠血清凝集紊水平的作用，结论蓝桉果在调动机体免疫系统机能方面有一定的促进作用．

凝集素化昔萘酸沙美特罗纳米粒-微粒的药效和药动学研究

目的研究小鼠体内干粉吸入麦胚凝集素(WGA)化修饰的昔萘酸沙美特罗(SalX)纳米粒-微粒(NiMS)的药效学及药动学。方法通过皮下和腹腔注射卵清蛋白(OVA)建立小鼠肺哮喘病理模型,经气道干粉吸入给予WGA-SalXNiMS粉末,采用对照实验法,检测给药后的肺组织及气道炎症情况,并对药物在小鼠血浆和肺组织中的药代动力学行为进行研究。数据用Sigma State统计软件包处理。结果与模型组相比,SalX-NiMS组和WGA-SalX-NiMS组小鼠支气管肺泡灌洗液(BALF)中白细胞总数、嗜酸粒细胞、淋巴细胞及巨噬细胞数目呈下降趋势。WGA-SalX-NiMS组小鼠BALF中淋巴细胞及巨噬细胞数与SalX-NiMS组相比减少(P<0.05)。在小鼠血浆中,SalX-NiMS组的tmax为1.500h,Cmax为57.366mg/L,t1/2β为69.315h,AUC0-∞为2 427.205mg·L-1·h,MRT0-∞为55.294h;WGA-SalX-NiMS组的tmax为1.000h,Cmax为62.581mg/L,t1/2β为69.315h,AUC0-∞为4 071.838mg·L-1·h,MRT0-∞为75.094h。在小鼠肺组织中,SalXNiMS组的tmax为0.083h,Cmax为0.497μg/mg,t1/2β为11.231h,AUC0-∞为3.936μg·mg-1·h,MRT0-∞为13.854h;WGASalX-NiMS组的tmax为0.083h,Cmax为0.796μg/mg,t1/2β为27.294h,AUC0-∞为5.578μg·mg-1·h,MRT0-∞为26.330h。WGA-SalX-NiMS组的血浆及肺组织药物浓度均高于SalX-NiMS组(P<0.05)。结论 WGA修饰后的SalX-NiMS在小鼠体内易于释药,血浆和肺组织中药物浓度较高,有利于哮喘发作时炎症的控制与改善。

多学科甲胎蛋白异质体临床应用专家共识

甲胎蛋白（Alpha-fetoprotein,AFP）是目前临床应用最广泛的肝癌辅助诊断血清标志物。AFP诊断肝癌的敏感性约为60%~70%。但在肝脏良性疾病,特别是肝癌高危人群,如慢性肝炎、肝硬化中也有部分患者会出现AFP升高。妇女孕期及某些生殖系统疾病AFP也会升高。

唾液酸受体在家禽肾和生殖系统组织中的表达

为了观察流感病毒唾液酸α2,3-半乳糖和唾液酸α2,6-半乳糖两种受体在鸡、鸭两种家禽肾脏和生殖系统组织中分布和表达的特征,本研究利用凝集素免疫荧光组织染色方法对各器官组织进行了系统的定性分析。结果显示,唾液酸α2,3-半乳糖和唾液酸α2,6-半乳糖这两种受体在鸡、鸭肾脏和生殖系统中皆有分布,其中唾液酸α2,3-半乳糖的分布更为广泛。这一研究结果提示,鸡、鸭肾脏和生殖系统组织细胞对禽流感病毒和人流感病毒都存在易感的基础,且更容易感染禽源的流感病毒。