Wound healing activity of alcoholic extract of Kaempferia galanga in Wistar rats

The wound healing effect of alcoholic extract of Kaempferia galanga (K. galanga) and its effect in dexamethasone suppressed wound healing was studied in Wistar rats. Three wound models viz. incision, excision and dead space wounds were used in this study. The parameters studied were breaking strength in case of incision wounds, epithelialization and wound contraction in case of excision wound and granulation tissue dry weight, breaking strength and hydroxyproline content in case of dead space wound. The dexamethasone treated group showed a significant (P < 0.001) reduction in the wound breaking strength when compared to control group in incision type of wound model. Coadministration of K. galanga with dexamethasone had significantly (P < 0.001) increased the breaking strength of dexamethasone treated group. In excision wound model, the percentage of the wound contraction was significantly (P < 0.05) increased by K. galanga only on 16th day and also it reversed the dexamethasone suppressed wound contraction on the 16 day. K. galanga significantly (P < 0.001) reduced the time required for epithelialization and reversed the epithelialization delaying effect of dexamethasone significantly (P < 0.001).     

Mature CD83+ dendritic cells infected with recombinant gp 100 vaccinia virus stimulate potent antimelanoma T cells

Background Mature dendritics cells (DCs) are potent antigen-presenting cells that activate naive T lymphocytes and initiate cellular immune responses. The ability of CD83⁺ mature DCs infected with vaccinia virus encoding the gp 100 melanoma transgene (rV-gp 100) to stimulate an antimelanoma CD8⁺ T-cell response was investigated. Methods Monocyte-derived immature or CD83⁺ mature DCs were infected with rV-gp100. The activation state of the DCs and the expression of gp 100 protein were evaluated by flow cytometry. The reactivity of antimelanoma CD8⁺ T cells was confirmed by measuring specific interferon γ secretion by using enzyme-linked immunosorbent assay in a mixed-tumor lymphocyte culture. Results Both immature and CD83⁺ mature DCs expressed gp 100 protein when the DCs were infected with rV-gp 100. Calcium-signaling agents were required to induce maturation of both infected and nonifected immature DCs. Only rV-gp100-infected CD83⁺ DCs induced CD8⁺ T cells, after a single stimulation that recognized both peptide-pulsed target cells to multiple gp 100 epitopes and a melanoma cell line that endogenously expressed gp 100 antigen. Conclusions CD83⁺ DCs transduced with rV-gp 100 are capable of generating a strong CD8⁺ T-cell response against melanoma tumor cells. Expression of melanoma antigens by mature DCs offers the potential advantage of presenting multiple endogenously processed T-cells epitopes and using multiple HLA restriction elements for antimelanoma vaccine therapy.     

Novel scintillation proximity assay for measuring membrane-associated steps of peptidoglycan biosynthesis in Escherichia coli

We have developed a novel, high-throughput scintillation proximity assay to measure the membrane-associated steps (stages 2 and 3) of peptidoglycan synthesis in Escherichia coli. At least five enzymes are involved in these two stages, all of which are thought to be essential for the survival of the cell. The individual enzymes are difficult to assay since the substrates are lipidic and difficult to isolate in large quantities and analysis is done by paper chromatography. We have assayed all five enzymes in a single mixture by monitoring synthesis of cross-linked peptidoglycan, which is the final product of the pathway. E. coli membranes are incubated with the two sugar precursors, UDP-N-acetyl muramylpentapeptide and UDP-[(3)H]-N-acetylglucosamine. The radiolabel is incorporated into peptidoglycan, which is captured using wheat germ agglutinin-coated scintillation proximity assay beads. The assay monitors the activity of the translocase (MraY), the transferase (MurG), the lipid pyrophosphorylase, and the transglycosylase and transpeptidase activities of the penicillin-binding proteins. Vancomyin, tunicamycin, nisin, moenomycin, bacitracin, and penicillin inhibit the assay, and these inhibitors have been used to validate the assay. The search for new antimicrobial agents that act via the late stages of peptidoglycan biosynthesis can now be performed in high throughput in a microtiter plate.     

Acidosis plus melphalan induces nitric oxide-mediated tumor regression in an isolated limb perfusion human melanoma xenograft model

BACKGROUND: Isolated limb perfusion (ILP) with melphalan is an accepted treatment for intransit melanoma of the extremities. Using an ILP human melanoma xenograft model, we tested the hypothesis that acidosis augments the antitumor effect of melphalan and that nitric oxide (NO) induction mediates tumor regression. METHODS: NIH1286 human melanoma tumor bearing athymic nude rats underwent a 10-minute ILP. Group C was perfused at physiologic pH without acid or melphalan, group M received melphalan at physiologic pH (7.2), group A received 0.2 N of HCl at pH 6.8, and group A/M received melphalan and HCl at pH 6.8. Groups 1400W + A and 1400W + A/M were injected with 1400W, a specific inhibitor of inducible NO synthase, 1 hour pre-ILP. Tumor response was followed for up to 60 days in all survival experiments. In 4 to 6 animals from groups C, M, A, and A/M, tumor NO was measured pre- and post-ILP, and tumor and thigh muscle from 2 additional animals in each group were collected at 20 minutes and 24 hours post-ILP and processed for terminal deoxynucleotidyl transferase dUTP nick end labeling staining. RESULTS: Maximum mean reduction in tumor size after ILP in the different groups was as follows: C = 0%, M = 55%, A = 99.6% (3 of 4 complete responses), A/M = 100% (all complete responses), 1400W + A = 0%, and 1400W + A/M = 25%. Median tumor NO was 0.87 +/- 0.74 (SD) micromol/L before ILP and increased significantly (Mann-Whitney rank sum test, P <.001) after ILP (C = +6.9%, n = 4; M = +7.5%, n = 5; A = +66.0%, n = 6; A/M = +35.9%, n = 6). Also, minimal apoptotic cell death was seen in C and M, whereas A and A/M showed evidence of widespread apoptosis. CONCLUSIONS: Acidosis enhances the antitumor effect of melphalan. NO induction appears to play a role in tumor regression.     

Virus-mediated oncolysis of thyroid cancer by a replication-selective adenovirus driven by a thyroglobulin promoter-enhancer region

CONTEXT: Currently, there is no effective treatment for iodine-resistant thyroid cancers. OBJECTIVE: As a new approach to treatment, the efficacy of replication-selective, human thyroglobulin (TG) enhancer and promoter-driven, adenovirus (AdhTGEP)-mediated oncolysis was investigated using two well-differentiated thyroid cancer cell lines, XTC (TG positive) and FTC-133 (TG negative), and other control tumor and nontumor cell lines (all TG negative). DESIGN: A cohort study design was used. SETTING: The study setting was laboratory bench-top experiments. SUBJECTS/PARTICIPANTS: In vitro TG-expressing and nonexpressing thyroid cell culture lines, nonthyroid tumor cell lines, as well as preclinical thyroid tumor-bearing mice were studied. INTERVENTION: Adenoviral infection of cell lines was determined by immunohistochemistry, selective replication by one-step growth assays, and cytotoxicity by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulphophenyl)-2H-tetrozolium (MTS) assay. In vivo tumor growth inhibition was determined by a single intratumoral injection of 1 x 10(9) plaque-forming units AdhTGEP, AdLacZ (control virus), or PBS to 50- to 75-mm(3) tumors. XTC cells showed intense immunohistochemical staining, whereas FTC-133 and all other control cell lines showed minimal staining for viral infection with AdhTGEP. MAIN OUTCOME MEASURES: Cell survival and tumor growth inhibition after adenoviral infection were the main outcome measures. RESULTS: One-step growth assays showed at least a more than 60-fold titer of AdhTGEP in XTC than in FTC-133 cells. Cytotoxicity assays showed approximately 68% cell kill in XTC and minimal cell kill in FTC-133 and all other control cell lines at a multiplicity of infection of 250. There was significant in vivo growth inhibition of AdhTGEP-treated XTC tumors (67 +/- 49 mm(3)) compared with AdLacZ-treated XTC (228 +/- 45 mm(3); P < 0.01), PBS-treated XTC (372 +/- 70 mm(3); P < 0.001), or AdhTGEP-treated FTC-133 tumors (598 +/- 168 mm(3)). CONCLUSION: Replication-selective virus-mediated oncolysis is a potential therapy for recurrent, well-differentiated, TG-secreting thyroid cancer that is unresponsive to standard treatment.     

A programme to eliminate lymphatic filariasis in Tamil Nadu state, India: compliance with annual single-dose DEC mass treatment and some related operational aspects

This paper reports on DEC distribution and compliance with treatment in a large-scale annual single-dose mass treatment programme to eliminate lymphatic filariasis in the south Indian state of Tamil Nadu. 76.9% of households (82.5% in rural areas and 58.0% in urban areas) were aware of drug distribution for control of filariasis. DEC was given to 70% (= distribution rate) (range 0-92%) of the population and 53.5% (range 12-89%) complied with treatment. The distribution rate was more than 75% in 74% of the villages and compliance was in the range of 51-75% in 76% of the villages. About 5% of the treated population reported side-effects. Distribution and compliance were higher in rural than urban areas and similar between males and females. Qualitative data showed that some socio-economic factors, logistic and drug-related problems and people's poor knowledge and perceived benefits of treatment played a role in a proportion of the population not receiving or taking the drug. The Tamil Nadu programme showed that large-scale repeated annual DEC mass treatment is feasible and that existing health services are capable of delivering the drug to all communities. While even poor to moderate compliance rates can reduce the vector transmission of infection to some extent, improved drug distribution and compliance with treatment are necessary to consolidate the gains of earlier rounds of treatment and achieve the goal of filariasis elimination within a reasonable time frame.