This study aimed to characterize the morphological changes in the ovary of the female crab Neptunus pelagicus and to identify specific fractions of vitelloginin and vitelline molecules during primary and secondary vit...This study aimed to characterize the morphological changes in the ovary of the female crab Neptunus pelagicus and to identify specific fractions of vitelloginin and vitelline molecules during primary and secondary vitellogenesis.Samples of the blue crab were collected from the Mediterranean Sea of Alexandria monthly during 2017.Ovaries and oocytes in primary and secondary vitellogenesis were detached and treated for histological test.Native polyacrylamide gel electrophoresis(PAGE)Bis-Tris Gels was applied to identify vitelloginin(VN)and vitelline(VL)molecules.Protein Analyses were done by PAGE-SDS.The initial degenerate primers were built regarding the conserved amino acid domains of the yolk proteins.Primary and secondary vitellogeneses consisted of 8 phases.Lipoprotein fraction with molecular weight 550 kDa was identified in the hemolymph in secondary vitellogenesis.Two protein fractions(VLI&VLII)were identified in secondary vitellogenic oocytes.The electrophoresis performed with extract of stage I oocyte showed two protein fractions with molecular weights 550 kDa and 460 kDa.In stage II and III oocyte,4 subunits were presented of 180,195,140 and 120 kDa in VLI and 2 subunits with molecular weight of 110 kDa and 95 kDa in VLII.Another two fractions in stage V oocyte presented with molecular weights of 380 kDa and 360 kDa.Western blot analysis proved that both fractions were of four major polypeptide subunits with molecular weight of 180,125,90 and 85 kDa in each of the two VLs.The hybridization signal obtained by the Northern blot was detected in the hepatopancreas during ovarian cycle and in the ovary during secondary vitellogenesis.The result of the reverse transcription-polymerase chain reaction(RT-PCR)analysis showed that the mRNA that encodes the C-terminal region of the VN cDNA was found in the ovary in secondary vitellogenesis and in the hepatopancreas.展开更多
Objective To comprehend the reiation of tobacco glycoprotein (TGP) to Buerger,s disease.Metbods TGP was isolated from crude tobacco leaves by basic immunologic techniques. Serum anti- TGPantibodies were tested by West...Objective To comprehend the reiation of tobacco glycoprotein (TGP) to Buerger,s disease.Metbods TGP was isolated from crude tobacco leaves by basic immunologic techniques. Serum anti- TGPantibodies were tested by Western blot analysis in 11 patients with Buerger,s disease, 15 healthy male smokers and11 nonsmoking healthy male subjects. Results 1. TGP is a dark brown protein of molecular weight 14000. It maybe a subunit of some high molecular weight protein, and exists in crude tobacco leaves. 2. Western blot analysisshowed that 81.81% of patients with Buerger’s disease (9/11), 33.33% of healthy smokers (5/15) and 27.27% ofhealthy nonsmokers (3/11) had serum anti- TGP antibodies. There was significant dtherence between patientswith Buerger,s disease and two control groups (P<0.05), and no signilicant dtherence between both control groups(P>0.05). Conclusion TGP does play an important role in the pathogenesis of Buerger’s disease. As anti - TGPantibodies are also found in some control subjects, it is speculated that other etiologic factors might coordinatelycontribute to the specifc vascular response to TGP in susceptible subjects.展开更多
文摘This study aimed to characterize the morphological changes in the ovary of the female crab Neptunus pelagicus and to identify specific fractions of vitelloginin and vitelline molecules during primary and secondary vitellogenesis.Samples of the blue crab were collected from the Mediterranean Sea of Alexandria monthly during 2017.Ovaries and oocytes in primary and secondary vitellogenesis were detached and treated for histological test.Native polyacrylamide gel electrophoresis(PAGE)Bis-Tris Gels was applied to identify vitelloginin(VN)and vitelline(VL)molecules.Protein Analyses were done by PAGE-SDS.The initial degenerate primers were built regarding the conserved amino acid domains of the yolk proteins.Primary and secondary vitellogeneses consisted of 8 phases.Lipoprotein fraction with molecular weight 550 kDa was identified in the hemolymph in secondary vitellogenesis.Two protein fractions(VLI&VLII)were identified in secondary vitellogenic oocytes.The electrophoresis performed with extract of stage I oocyte showed two protein fractions with molecular weights 550 kDa and 460 kDa.In stage II and III oocyte,4 subunits were presented of 180,195,140 and 120 kDa in VLI and 2 subunits with molecular weight of 110 kDa and 95 kDa in VLII.Another two fractions in stage V oocyte presented with molecular weights of 380 kDa and 360 kDa.Western blot analysis proved that both fractions were of four major polypeptide subunits with molecular weight of 180,125,90 and 85 kDa in each of the two VLs.The hybridization signal obtained by the Northern blot was detected in the hepatopancreas during ovarian cycle and in the ovary during secondary vitellogenesis.The result of the reverse transcription-polymerase chain reaction(RT-PCR)analysis showed that the mRNA that encodes the C-terminal region of the VN cDNA was found in the ovary in secondary vitellogenesis and in the hepatopancreas.
文摘Objective To comprehend the reiation of tobacco glycoprotein (TGP) to Buerger,s disease.Metbods TGP was isolated from crude tobacco leaves by basic immunologic techniques. Serum anti- TGPantibodies were tested by Western blot analysis in 11 patients with Buerger,s disease, 15 healthy male smokers and11 nonsmoking healthy male subjects. Results 1. TGP is a dark brown protein of molecular weight 14000. It maybe a subunit of some high molecular weight protein, and exists in crude tobacco leaves. 2. Western blot analysisshowed that 81.81% of patients with Buerger’s disease (9/11), 33.33% of healthy smokers (5/15) and 27.27% ofhealthy nonsmokers (3/11) had serum anti- TGP antibodies. There was significant dtherence between patientswith Buerger,s disease and two control groups (P<0.05), and no signilicant dtherence between both control groups(P>0.05). Conclusion TGP does play an important role in the pathogenesis of Buerger’s disease. As anti - TGPantibodies are also found in some control subjects, it is speculated that other etiologic factors might coordinatelycontribute to the specifc vascular response to TGP in susceptible subjects.
