Intracellular Localization of HBV Capsid in Hepatocyte Line After Transfected by The Entire HBV Genome = Lokalisasi Intraseluler Kapsid HBV Pada Sel Line Hepatosit Setelah Ditransfeksi Dengan Genom Utuh HBV.

HBV replicates within the nucleus of hepatocyte using cellular transport machinery for the import of their genomes into the nucleus. Genome of HBV has to transported through the cytoplasm towards the nuclear pore complex (NPC) followed by subsequent passage through the pore. HBV capsid is involved in a number of important functions in the replication cycle of HBV. It can be detected in the nucleus, cytoplasm or both within infected hepatocytes. Nuclear localization of HBV capsid protein, which is karyophilic, depends on the cell cycle. The objective of the present study was to analyzes the intracellular localization of HBV capsid protein after transfected by entire HBV genome into hepatocyte cell lines (HuH-7) and to determine the predominantly localization of the capsid into cell compartment. In this work we analysed the intracellular localization of the HBV capsid in human hepatocyte cell lines liuH-7 by transfection using entire HBV genome and transient expression. The transfected cells were fixed and an indirect immune staining against the HBV capsid was performed to detect the capsid. To verify the location within the cell, an additional co-staining against the nuclear pore complexes was performed. The Intracellular localization of the HBV capsid and NPC were analyzed by a confocal laser scan microscope. The observed of localizations into the transfected cells were classified to be predominantly as nuclear localization, cytoplasmic localization or distributed within both of these compartments. Result of this study indicated that Staining of HBV capsid found predominantly within the nucleus (71%). Less frequently, the HBV capsid localized within the cytoplasm (26%). Only in a minority of cases, the capsids were localized within cytoplasm and nucleus (3%). This low frequency indicate that the capsids were not diffusing within the cells being in accordance to the in vivo situation in which the nuclear membrane was impermeable for the capsid. Key words: HBV capsid, hepatocyte, NPC, HuH-7, confocal laser scan microscope Replikasi HBV terjadi di dalam nukleus hepatosit dengan mempergunakan mekanisme transport seluler untuk mengirimkan genomnya ke dalam nukleus. Genom HBV hams ditransportasikan ke dalam nukleus melalui nuclear pore complex (NPC). Kapsid HBV terlibat dalam sejumlah fungsi panting dalam siklus replikasi HBV. Protein kapsid HBV dapat dideteksi didalam nukleus, sitoplasm ataupun di kedua kompartemen tersebut di dalam sel hepatosit yang terinfeksi HBV. Lokalisasi protein kapsid HBV yang bersifat karyophilik tergantung pada siklus sel. Adapun tujuan dari penelitian ini adalah untuk menganalisis lokalisasi intraseluler protein kapsid HBV pada sel line hepatosit HuH-7 pasca ditransfeksi dengan genom HBV utuh serta untuk menentukan lokalisasi kapsid HBV yang dominan dalam kompartemen sel. Pada penelitian telah dilakukan analisis lokalisasi intraseluler kapsid HBV dengan mentransfeksikan genome HBV utuh dan mengekspresikannya di dalam sel line HuH-7. Sel yang sudah ditransfeksi selanjumya difiksasi dan dilakukan penwarnaan secara tidak langsung terhadap kapsid HBV. Untuk menentukan lokalisasi dalam sel, pewarnaan lanjutan dilakukan terhadap NPC. Selanjutnya, dengan menggunakan mikroskop scan laser konfokal dianalisis lokalisasi intraseluler kapsid HBV dan NPC. Hasil pengamatan menunjukan bahwa distribusi HBV di dalam nukleus lebih dominan dibandingkan dengan distribusi di dalam sitoplasma dan distribusi pada kedua kompartemen tersebut. Secara kuantitatif menunjukkan bahwa distribusi protein kapsid HBV dalam nukleus adalah dominan dengan 71%, sedangkan dalam sitoplasma adalah 26%, sangat minoritas dijurnpai distribusi protein kapsid HBV pada kedua kompartemen sel, yaitu hanya 3%. Hal ini menunjukkan bahwa masuknya kapsid HBV kedalam nukleus tidak terdifusi secara pasif, namun secara aktif melalui mekanisme protein transport melalui NPC. Membran nukleus bersifat permeable terhadap protein HBV kapsid. Kata kunci: kapsid HBV, hepatosit, NPC, HuH-7, mikroskop scan laser konfoca

Effect of Staurosporine on the Intracellular Localization of Hepatitis B Virus Core Protein

protein is also including in the HBV genome targeting into the nucleus through modulating carboxyl residues byphosphorylation. Nuclear localication Signal (NLS) in HBV core protein is inside the virion structure and it must beunmasked in order to function, perhaps by phosphorylation. Phosphorylation of of HBV core protein in turn couldbegin to alter capsid conformation. Staurosporine is a natural product originally isolated from bacteriumStreptomyces staurosporeus. Staurosporine was discovered to have biological activities ranging from anti-fungal toanti-hypertensive. The interest in these activities resulted in a large investigative effort in chemistry and biology andthe discovery of the potential for anti-cancer treatment. The main biological activity of Staurosporine is the inhibitionof protein kinases through the prevention of ATP binding to the kinase. In the present study, we have studied theintracellular localization of EGFP-Core fusion protein with triple HBV core and SV-40 nuclear localization signal atits carboxyl terminal in presence and absence of Staurosporine. We also to study the effect of Staurosporine treatmenton the intracellular localization of EGFP-Core fusion protein in the hepatocyte cells line of HepG2 cell. Resultsshowed that effect of Staurosporine is prevent the nuclear localization of EGFP-Core fusion protein into nucleusthrough an inhibition of the phosphorylation of core protein. Stauroporine also prevents cell division so that passivetrapping of core protein is inhibited

Effect of Nuclear Export Inhibitor Leptomycin Bon the Intracellular Localization of HBV Core Protein into Hepatocytes Cell Line Huh-7 and HepG2 Cells

Leptomycin B (LMB) was originally discovered as a potent anti-fungal antibiotic from Streptomyces species. The cellular target of LMBhas been identified as the nuclear export receptor CRM-1 or exportin-1, which is involved in nuclear trafficking of cellular RNAs or proteins containing the nuclear export sequence (NES). CRM-1 is the main mediator of nuclear export in many cell types including hepatocyte cell lines. The ability of LMB to inhibit nuclear export has made it a useful tool in the study of the intracellular localization of many regulatory proteins. In this study, we evaluated the effect of nuclear export inhibitor LMB treatment on the intracellular localization of HBV core protein into the hepatocyte cell lines, Huh-7 and HepG2 cells. We also reported the quantification of the distribution of EGFP-Core fusion protein with redundant core NLS as well as SV-40 NLS into cell compartments. Results shown that in Huh-7 cells treatment of LMBcaused retention of EGFP-Core fusion protein into the nucleus, so increased the nuclear localization of EGFP-Core and all variants. In HepG2 cells, although not significantly, treatment of LMB increased a number of nuclear localization in all EGFP-Core constructions, even the nuclear localization in HepG2 cells is not so high as in Huh-7 cells

Factorization at subleading power in deep inelastic scattering in the x→1x\rightarrow 1 limit

We examine the endpoint region of inclusive deep inelastic scattering at next-to-leading power (NLP). Using a soft-collinear effective theory approach with no explicit soft or collinear modes, we discuss the factorization of the cross section at NLP and show that the overlap subtraction procedure introduced to eliminate double counting of degrees of freedom at leading power ensures that spurious endpoint divergences in the rate cancel at NLP at one loop. For this cancellation to occur at all renormalization scales a nontrivial relation between the anomalous dimensions of the leading and subleading operators is required, which is demonstrated to hold at one loop.Comment: 9 pages, 1 figure; typos corrected, words added, references adde

Effect of Nuclear Export Inhibitor Leptomycin B on the Intracellular Localization of HBV Core Protein into Hepatocytes Cell Line Huh-7 and HepG2 Cells

Leptomycin B (LMB) was originally discovered as a potent anti-fungal antibiotic from Streptomyces species. The cellular target of LMB has been identified as the nuclear export receptor CRM-1 or exportin-1, which is involved in nuclear trafficking of cellular RNAs or proteins containing the nuclear export sequence (NES). CRM-1 is the main mediator of nuclear export in many cell types including hepatocyte cell lines. The ability of LMB to inhibit nuclear export has made it a useful tool in the study of the intracellular localization of manyregulatory proteins. In this study, we evaluated the effect of nuclear export inhibitor LMB treatment on the intracellular localization of HBV core protein into the hepatocyte cell lines, Huh-7 and HepG2 cells. We also reported the quantification of the distribution of EGFP-Core fusion protein with redundant core NLS as well as SV-40 NLS into cell compartments. Results shown that in Huh-7 cells treatment of LMB caused retention of EGFP-Core fusion protein into the nucleus, so increased the nuclear localization of EGFP-Core and all variants.In HepG2 cells, although not significantly, treatment of LMB increased a number of nuclear localization in all EGFP-Core constructions, even the nuclear localization in HepG2 cells is not so high as in Huh-7 cells. Keywords: Leptomycin B, HBV, core protein, intracellular localization, NLS, Huh-7, HepG2 cel

INKULTURASI PASKAH DI RANTEPAO, EKSPRESI IMAN DALAM BUDAYA TORAJA

Humans are homo symbolicum, the creatures who use sign/symbol to express a certain purpose. Almost human’s actions are expressed in symbols, including in religious life. One of the religious expressions that are interesting to study which showed by the Catholics in Rantepao is Easter Celebration. Therefore, this research especially studies the Catholic Inculturation and the local culture of Toraja that is actualized in the Easter Celebration in Rantepao. Rantepao Parish has tried to bring another Easter Celebration feel frame in Toraja nuances. Various local symbols were appointed to further enliven the Easter Celebration. The great celebration was expressed in the treasure of the thought of the Toraja people

Immunoglobulin Structure Exhibits Control over CDR Motion

Motions of the IgG structure are evaluated using normal mode analysis of an elastic network model to detect hinges, the dominance of low frequency modes, and the most important internal motions. One question we seek to answer is whether or not IgG hinge motions facilitate antigen binding. We also evaluate the protein crystal and packing effects on the experimental temperature factors and disorder predictions. We find that the effects of the protein environment on the crystallographic temperature factors may be misleading for evaluating specific functional motions of IgG. The extent of motion of the antigen binding domains is computed to show their large spatial sampling. We conclude that the IgG structure is specifically designed to facilitate large excursions of the antigen binding domains. Normal modes are shown as capable of com- putationally evaluating the hinge motions and the spatial sampling by the structure. The antigen binding loops and the major hinge appear to behave similarly to the rest of the structure when we consider the dominance of the low frequency modes and the extent of internal motion. The full IgG structure has a lower spectral dimension than individual Fab domains, pointing to more efficient information transfer through the antibody than through each domain. This supports the claim that the IgG structure is specifically constructed to facilitate antigen binding by coupling motion of the antigen binding loops with the large scale hinge motions