MOLECULAR ANALYSIS OF GENE EXPRESSION RELATED TO THE EFFECTS OF DLBS3233 TREATMENT IN DIFFERENTIATION OF 3T3-L1 PRE-ADIPOCYTE

Objective: DLBS3233 is a standardized extract combination containing Lagerstroemia speciosa and Cinnamomum burmannii. The effect of DLBS3233 on adipocyte differentiation was examined in this study.Methods: 3T3-L1 pre-adipocyte was used to investigate gene expression using the real-time reverse transcription polymerase chain reaction (RT-PCR) method. Oil red-O staining for detecting lipid formation was also carried out in this experiment.Results: DLBS3233 caused cell differentiation of 3T3-L1 pre-adipocytes into adipocytes which were indicated by positive results on staining cells with oil red-O on day 6 of the differentiation process. Analysis of gene expressions associated with adipogenesis (C/EBP-α, PPAR-γ, C/EBP-δ, FASn and adiponectin) showed an increase compared to control. In this study, DLBS3233 at a concentration of 5 μg/ml exhibited better differentiation effect than DLBS3233 at a concentration of 10 μg/ml.Conclusion: DLBS3233 can stimulate differentiation of 3T3-L1 pre-adipocytes into adipocytes.Keywords: DLBS3233, Adipogenesis, Gene expression analysis, Real-time RT-PC

Removing Cystein Group On Interferon Alpha 2b at Position 2 and 99 does Not Diminish Antitumor Activity of the Protein, Even Better

Interferon alpha 2b is the only standard therapeutic protein for hepatitis virus infections. Further study demonstrated that this protein also posseses antitumor activity in several cancerous organs. One main pathway of this antitumor activity is mediated through antiproliferation as well as proapoptotic effects. Previously, we have successfully developed recombinant human interferon alpha 2b (rhIFN alpha 2b) by using a synthetic gene. In addition, two mutein forms of rhIFN alpha 2b were generated to improve the characteristics of this protein. Two point mutations showed better pharmacokinetic profiles than one point mutation as well as the native form. In the present study, this mutein form was studied for ist antitumor effect in vitro using HepG2 cells. As a comparison, the native form as well as a commercial rIFN alpha 2b were used. Several parameters were investigated including the MTT assay, cell viability test, cell cycle using flow cytometric analysis, and the genes and protein expressions involved in cell growth. The latest was observed to study the mechanism of rhIFN alpha 2b. There was no significant difference in the MTT assay and cell viability after cells were treated with both forms of rhIFN alpha 2b. However, the mutein rhIFN alpha 2b tended to show better proapoptotic activity reflected by flow cytometric data, protein expression of pSTAT1, and DNA expression of caspase 3

THE EXISTENCE OF GENE DNA POLYMERASE MUTATION FROM POSITIVE HEPATITIS B SAMPLES IN BANDUNG, INDONESIA

Objective: To complete mutation existence in reverse transcriptase domain of the viral Polymerase.Methods: The study was done by amplification step of viral polymerase gene fragment, agarose gel electrophoresis, PCR product purification with GFX column kit, sequencing and sequencing result analysis. The samples were derived from Clinical Laboratory in Bandung Indonesia.Results: The result showed mutations in DNA fragment encoding for RT domain viral polymerase in sample 6, 7 and 8. There were mutations leading to amino acid substitution L526S in sample 6, D551E in sample 7 and D552E in sample 8.Conclusion: D551E and D552E substitution occurred in YMDD motif RT DNA polymerase that produced YMDE mutant. L526S, D551E and D552E were estimated as antivirus-resistance mutants that have never been reported before.  Â

Kloning gen lumbricin-1 dari lumbricus rubellus isolat Aceh serta overproduksi, purifikasi dan uji aktivitas antimikroba

viii.;43 hlm.;ilus.;tab.;29 cm

Evaluation of alum-based adjuvant on the immunogenicity of salmonella enterica serovar typhi conjugates vaccines

The function of adjuvant in maintaining the long-term immune response to Typhoid conjugate vaccine (TCV) was evaluated in. Two TCV products, Vi-DT and Vi-TT, were formulated in either aluminum phosphate (AlPO4) or aluminum hydroxide (AlOH) as adjuvants and TCV formulated in phosphate buffer saline were used as controls. In each case, a group of Balb/c mice was injected intramuscularly with two doses of the formulated vaccine at two-week intervals. The anti-Vi IgG responses were monitored by Enzyme-Linked Immunosorbent Assay and the levels of CD4+ T-cells expressing cytokine were characterized using intracellular cytokine staining. All mice immunized by TCV formulated in adjuvant elicited anti-Vi response to a higher level than the group receiving TCV formulated in PBS. The extent of adsorption of TCV in AlOH was greater than that in AlPO4, and this finding correlated well with the observation that the mice immunized with two doses of Vi-DT(AlOH) elicited anti-Vi IgG to a level higher than that seen with Vi-DT(AlPO4). The mice primed with Vi-TT(AlOH) produced lower anti-Vi IgG (25.901 GM) compared to those receiving Vi-TT(AlPO4) (49.219 GM). However, after the second injection, the former raised the antibody level significantly to 137.008 GM while the latter provided a value of only 104.966 GM. The groups of mice vaccinated by TCV formulated in AlOH expressed IL4 at higher levels than the other groups, which correlated positively with the high Anti-Vi IgG in these animals. In conclusion, AlOH could be recommended as an effective adjuvant for TCV to provide a long-term immune response

Development of Chimeric Hepatitis B (HBV) – Norovirus (NoV) P particle as candidate vaccine against Hepatitis B and norovirus infection

Introduction: Hepatitis B remains a global problem with no effective treatment. Here, a mucosal vaccine candidate was developed with HBsAg and HBcAg, to provide both prophylactic and therapeutic protection against hepatitis B. The antigens were presented using the P particle of human norovirus (HuNov). As a result, the chimeric HBV – HuNoV P particle can act as a dual vaccine for hepatitis B and HuNoV. Methods: The vaccine candidate was expressed and purified from Escherichia coli BL21 (DE3) cells. HBV-HuNoV chimeric P particles were successfully expressed and isolated, with sizes of approximately 25.64 nm. Then, the HBV-HuNoV chimeric P particles were evaluated for safety and immunogenicity in mice and gnotobiotic (Gn) pigs. After three doses (5 µg/dose in mice and 200 µg/dose in Gn pigs) of intranasal immunization, humoral and cellular immune responses, as well as toxicity, were evaluated. Results: The vaccine candidate induced strong HBV-HuNoV specific IFN-γ producing T-cell responses in the ileum, spleen, and blood of Gn pigs. Serum IgG and IgA antibodies against HBV-HuNoV chimeric P particles also increased significantly in Gn pigs. Increased HBsAg- and HuNoV-specific serum IgG responses were observed in mice and Gn pigs, although not statistically significant. The vaccine candidate did not show any toxicity in mice. Conclusions: In summary, the chimeric HBV-HuNoV P particle vaccine given intranasally was safe and induced strong cellular and humoral immune responses in Gn pig. Modifications to the vaccine structure and dosage need to be evaluated in future studies to further enhance immunogenicity and induce more balanced humoral and cellular responses