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...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.展开更多
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 ...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.展开更多
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 fre...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.展开更多
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,...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.展开更多
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 ...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.展开更多
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...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.展开更多
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 lymphoprolifera...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.展开更多
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 detec...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...展开更多
<strong>Background:</strong> 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 wit...<strong>Background:</strong> 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. <strong>Methodology:</strong> 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. <strong>Results:</strong> 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. <strong>Conclusion:</strong> 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.展开更多
<strong>Background:</strong> 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 aggl...<strong>Background:</strong> 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. <strong>Methods and Materials:</strong> 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<span style="white-space:nowrap;">°</span>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<span style="white-space:nowrap;">°</span>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 (<em>p</em> < 0.01). <strong>Results:</strong> 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. <strong>Conclusions:</strong> 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.展开更多
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 speci...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.展开更多
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 a...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.展开更多
Arisaema heterophyllum Blume is one of the three medicinal plants known as traditional Chinese medicine Rhizoma Arisaematis(RA). RA has been popularly used to treat patients with convulsions, inflammation, and cancer ...Arisaema heterophyllum Blume is one of the three medicinal plants known as traditional Chinese medicine Rhizoma Arisaematis(RA). RA has been popularly used to treat patients with convulsions, inflammation, and cancer for a long time. However, the underlying mechanisms for RA effects are still unclear. The present study was designed to determine the cytotoxicity of agglutinin isolated from Arisema heterophyllum Blume(AHA) and explore the possible mechanisms in human non-small-cell lung cancer A549 cells. AHA with purity up to 95% was isolated and purified from Arisaema heterophyllum Blume using hydrophobic interaction chromatography. AHA dose-dependently inhibited the proliferation of A549 cells and induced G_1 phase cell cycle arrest. AHA induced apoptosis by up-regulating pro-apoptotic Bax, decreasing anti-apoptotic Bcl-2, and activating caspase-9 and caspase-3. In A549 cells treated with AHA, the PI3K/Akt pathway was inhibited. Furthermore, AHA induced increase in the levels of ER stress markers such as phosphorylated eukaryotic initiation factor 2α(p-eIF2α), C/EBP-homologous protein(CHOP), inositol-requiring enzyme 1α(IRE1α), and phosphorylated c-Jun NH_2-terminal kinase(p-JNK). AHA also induced autophagy in A549 cells. Staining of acidic vesicular organelles(AVOs) and increase in the levels of LC3II and ATG7 were observed in AHA-treated cells. These findings suggested that AHA might be one of the active components with anti-cancer effects in Arisaema heterophyllum Blume. In conclusion, cytotoxicity of AHA on cancer cells might be related to its effects on apoptosis and autophagy through inhibition of PI3K/Akt pathway and induction of ER stress.展开更多
In order to find new insect resistance genes, four homologous cDNAs, hta-a, hta-b, hta-c and hta-d with lengths of 775, 718, 784 and 752 bp, respectively (GenBank accession numbers AF477031-AF477034), were iso...In order to find new insect resistance genes, four homologous cDNAs, hta-a, hta-b, hta-c and hta-d with lengths of 775, 718, 784 and 752 bp, respectively (GenBank accession numbers AF477031-AF477034), were isolated from a tuber cDNA expression library of Helianthus tuberosus L. Sequence analysis revealed that all four cDNAs contain an open reading frame of 444 bp, coding a polypeptide of 147 amino acid residues, and that the sequences of the cDNAs are very similar to those of the mannose-binding agglutinin genes of the jacalin-related family. In hemagglutination reactions and hapten inhibition assays, affinity-purified HTA (Helianthus tuberosus agglutinin) from induced Escherichia coli BL21(DE3) expressing GST-HTA shows hemagglutination ability and a higher carbohydrate-binding ability for mannose than other tested sugars. Trypsin inhibitory activity was detected in the crude extracts of induced E. coli BL21(DE3) expressing HTA, and was further verified by trypsin inhibitory activity staining on native polyacrylamide gel. The mechanism of interaction between HTA and trypsin was studied by molecular modeling. We found that plenty of hydrogen bonds and electrostatic interactions can be formed between the supposed binding sites of HTA-b and the active site of trypsin, and that a stable HTA/trypsin complex can be formed. The results above imply that HTA might be a bifunctional protein with carbohydrate-binding activity and trypsin inhibitory activity. Moreover, Northern blotting analysis demonstrated that hta is predominantly expressed in tubers of H. tuberosus, very weakly expressed in stems, but not expressed at all in other tissues. Southern blotting analysis indicated that hta is encoded by a multi-gene family. The insect resistance traits have been described in another paper.展开更多
In order to enhance the resistance to pests, transgenic maize (Zea mays L.) plants from elite inbred lines containing the gene encoding snowdrop lectin (Galanthus nivalis L. agglutinin; GNA) under control of a phloem-...In order to enhance the resistance to pests, transgenic maize (Zea mays L.) plants from elite inbred lines containing the gene encoding snowdrop lectin (Galanthus nivalis L. agglutinin; GNA) under control of a phloem-specific promoter were generated through the Agrobacterium tumefaciens-mediated also studied. Thirty-six independently derived plants were subjected to molecular analyses. The level of GNA expression at 0.13%-0.28% of total soluble protein was observed in different transgenic plants. The progeny of three GNA-expressing independent transformants that were derived separately from the elite inbred lines DH4866, DH9942, and 8902, were selected for examination of resistance to ACB. These plants synthesized GNA at levels above 0.24% total soluble protein and enhanced resistance to ACB was demonstrated by exposing the plants to insects under greenhouse conditions. Semi-artificial diet bioassays also showed the toxic effect of GNA on ACB. Field evaluation of the transgenic plants supported the results from the artificial trial. In the present study, we have obtained new insect-resistant maize material for further breeding work and have found that GNA-expressing plants not only gained significant resistance to homopterans, but also showed toxicity to ACB, which is a type of Lepidoptera.展开更多
文摘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.
文摘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.
文摘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.
基金supported by the National NaturalScienceFoundationofChina(No.31802093)Fund Project of Jilin Agricultural University(2019019).
文摘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.
文摘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.
文摘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.
文摘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.
文摘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...
文摘<strong>Background:</strong> 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. <strong>Methodology:</strong> 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. <strong>Results:</strong> 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. <strong>Conclusion:</strong> 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.
文摘<strong>Background:</strong> 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. <strong>Methods and Materials:</strong> 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<span style="white-space:nowrap;">°</span>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<span style="white-space:nowrap;">°</span>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 (<em>p</em> < 0.01). <strong>Results:</strong> 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. <strong>Conclusions:</strong> 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.
文摘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.
文摘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.
基金supported by the National Nature Science Foundation of China(No.81373964)Shanghai Science&Technology Support Program(No.13431900401)+1 种基金Shanghai Science&Technology Innovation Action Program(No.15140904800)the National Science&Technology Major Project of China(No.2014 ZX09301-306-03)
文摘Arisaema heterophyllum Blume is one of the three medicinal plants known as traditional Chinese medicine Rhizoma Arisaematis(RA). RA has been popularly used to treat patients with convulsions, inflammation, and cancer for a long time. However, the underlying mechanisms for RA effects are still unclear. The present study was designed to determine the cytotoxicity of agglutinin isolated from Arisema heterophyllum Blume(AHA) and explore the possible mechanisms in human non-small-cell lung cancer A549 cells. AHA with purity up to 95% was isolated and purified from Arisaema heterophyllum Blume using hydrophobic interaction chromatography. AHA dose-dependently inhibited the proliferation of A549 cells and induced G_1 phase cell cycle arrest. AHA induced apoptosis by up-regulating pro-apoptotic Bax, decreasing anti-apoptotic Bcl-2, and activating caspase-9 and caspase-3. In A549 cells treated with AHA, the PI3K/Akt pathway was inhibited. Furthermore, AHA induced increase in the levels of ER stress markers such as phosphorylated eukaryotic initiation factor 2α(p-eIF2α), C/EBP-homologous protein(CHOP), inositol-requiring enzyme 1α(IRE1α), and phosphorylated c-Jun NH_2-terminal kinase(p-JNK). AHA also induced autophagy in A549 cells. Staining of acidic vesicular organelles(AVOs) and increase in the levels of LC3II and ATG7 were observed in AHA-treated cells. These findings suggested that AHA might be one of the active components with anti-cancer effects in Arisaema heterophyllum Blume. In conclusion, cytotoxicity of AHA on cancer cells might be related to its effects on apoptosis and autophagy through inhibition of PI3K/Akt pathway and induction of ER stress.
基金Supported by the State Key Basic Research and Development Plan of China(G2000016205)the Hi-Tech Research and Development(863) Program of China(2001AA212041)+2 种基金the National Special Program for Research and Industrialization of Transgenic Plants(J99-B-004)the Rockefeller Foundation,the International Program on Rice Biotechnology(RF 97042 #14)the Special Supporting Program of the Chinese Academy of Sciences(STZ98-3-08)
文摘In order to find new insect resistance genes, four homologous cDNAs, hta-a, hta-b, hta-c and hta-d with lengths of 775, 718, 784 and 752 bp, respectively (GenBank accession numbers AF477031-AF477034), were isolated from a tuber cDNA expression library of Helianthus tuberosus L. Sequence analysis revealed that all four cDNAs contain an open reading frame of 444 bp, coding a polypeptide of 147 amino acid residues, and that the sequences of the cDNAs are very similar to those of the mannose-binding agglutinin genes of the jacalin-related family. In hemagglutination reactions and hapten inhibition assays, affinity-purified HTA (Helianthus tuberosus agglutinin) from induced Escherichia coli BL21(DE3) expressing GST-HTA shows hemagglutination ability and a higher carbohydrate-binding ability for mannose than other tested sugars. Trypsin inhibitory activity was detected in the crude extracts of induced E. coli BL21(DE3) expressing HTA, and was further verified by trypsin inhibitory activity staining on native polyacrylamide gel. The mechanism of interaction between HTA and trypsin was studied by molecular modeling. We found that plenty of hydrogen bonds and electrostatic interactions can be formed between the supposed binding sites of HTA-b and the active site of trypsin, and that a stable HTA/trypsin complex can be formed. The results above imply that HTA might be a bifunctional protein with carbohydrate-binding activity and trypsin inhibitory activity. Moreover, Northern blotting analysis demonstrated that hta is predominantly expressed in tubers of H. tuberosus, very weakly expressed in stems, but not expressed at all in other tissues. Southern blotting analysis indicated that hta is encoded by a multi-gene family. The insect resistance traits have been described in another paper.
文摘In order to enhance the resistance to pests, transgenic maize (Zea mays L.) plants from elite inbred lines containing the gene encoding snowdrop lectin (Galanthus nivalis L. agglutinin; GNA) under control of a phloem-specific promoter were generated through the Agrobacterium tumefaciens-mediated also studied. Thirty-six independently derived plants were subjected to molecular analyses. The level of GNA expression at 0.13%-0.28% of total soluble protein was observed in different transgenic plants. The progeny of three GNA-expressing independent transformants that were derived separately from the elite inbred lines DH4866, DH9942, and 8902, were selected for examination of resistance to ACB. These plants synthesized GNA at levels above 0.24% total soluble protein and enhanced resistance to ACB was demonstrated by exposing the plants to insects under greenhouse conditions. Semi-artificial diet bioassays also showed the toxic effect of GNA on ACB. Field evaluation of the transgenic plants supported the results from the artificial trial. In the present study, we have obtained new insect-resistant maize material for further breeding work and have found that GNA-expressing plants not only gained significant resistance to homopterans, but also showed toxicity to ACB, which is a type of Lepidoptera.