[Comparison of low voltage amplitude electrochemotherapy with 1 Hz and 5 kHz frequency in volume reduction of mouse mammary tumor in Balb/c Mice]

Electrochemotherapy [ECT] is an efficient technique that high intensity electric pulses in combination with chemotherapeutic drugs are applied to tumor cells. The most important unpleasant sensation of electrochemotherapy is muscle contraction. To resolve this problem, there are two solutions: first, increasing the repetition frequency of electric pulses above the tetanic frequency; and, second, reducing the voltage amplitude. ECT using 1 Hz or 5 kHz frequency at high amplitude examined and no difference response of the tumor treatment was observed. But the role of frequency in low amplitude ECT not examined. Therefore, the present study compared the anti-tumor effectiveness of electrochemotherapyusing electric pulses with frequencies of 1 Hz and 5 kHz at 70 v/cm amplitude. ECT of spontaneous mouse mammary tumor [SMMT] transplanted in Balb/c was performed with intratumoral injection of bleomycine and applied four different electric pulse protocols: 1- a train of 8 pulses with duration 50 ms, 1 Hz frequency and 70 v/cm amplitude, 2- a train of 4000 pulses with duration 100 micros, 5k Hz frequency and 70 v/cm amplitude,3- a train of 8 pulses with duration 100 micro s, 1 Hz frequency and 1000 v/cm amplitude,4- a train of 8 pulses with duration 100 micros, 5 kHz frequency and 1000 v/cm amplitude. Our data demonstrate significant differences in tumor volumes between mice treated by 70 V/cm and 5 kHz frequency compared to1 Hz frequency but inhibited tumor growth in high amplitude with 1 Hz and 5 kHz is comparable. Based on these results, the pulses with 70 v/cm and 5 kHz frequency are most effective. On the basis of these results the frequency effect of electric pulses is important in low amplitude ECT.

[Evaluation of cell proliferation and interleukine-4 and interferon gamma level measurement in Balb/c with experimental fibrosarcoma tumor by combination of Gp96-tumor peptide and naloxan]

In spite of the increasing progress in tumor treatment by current methods like surgery, chemotherapy and etc, medical sciences are unable to treat tumors. In this respect, immunology has opened a new window for tumor treatment; nowadays tumor immunotherapy is an accepted strategy for treatment of some tumors at least in some animal models. The goal of this study is the evaluation of immunotherapy using gp96- tumor peptide complex and its combination with naloxon as an opioid receptor antagonist to achieve of cellular immunity against tumors. In this study firstly, gp96 - tumor peptide complexes were purified from WEHI164 cells line using srivastava method. In the next stage, the mice, made tumoric before by the injection of tumor cells, then were divided in to four groups. Control group were injected by PBS, test group1 were injected by naloxon, test group2 were injected by gp96 - tumor peptide complex and test group3 were injected by combination of naloxon and gp96 - tumor peptide complex. To evaluation the efficacy of vaccination, after several days, tumor volume was recorded; then the mice were killed and the spleanic cells were extracted in sterile condition. MTT test was done for cells proliferation study. Supernatant of cultured cells were collected and assayed by ELISA kits for measuring IL-4 and IFN- gamma. Result of protein purification had showen, purified gp96 Isoform has Molecular Weight of 66 kilo dalton.Results of tumor volume had shown that, there is no significant difference between test and control groups. Results of MTT test had shown that, there is no significant difference between test and control groups. IL-4 assay study had showed that, there is no significant difference between test group1, group2 and control group but test group3 has significantly decreased in IL-4 amount when compared with control group. Results of IFN-gamma assay showed that, there is no significant difference between test group1 and control group, but test group2 and group3 has significantly increased in IFN- gamma amount when compared with control group. It can be concluded from this study is that, prophylactic immunotherapy of tumor by combination of gp96-tumor peptide complex and naloxon, can increase IFN- gamma, and, probably in a higher dosage, it may stimulate immune system more to become more potent to even decrease tumor volume

[Cloning and sequencing of L. major leishmania homologue of receptors for activated C kinase [LACK] gene]

Leishmaniasis is caused by parasitic protozoa of the genus Leishmania which in the infected host is obligate intracellular parasite. LACK gene is conserved among related Leishmania species. LACK is the immuno-dominant antigen of L.major which is considered as the most promising molecule for a recombinant or DNA vaccine against leishmaniasis. In this study the genomic DNA of an Iranian standard strain of Leishmania major [MRHO/IR/75/ER] was ertracted and the LACK gene was amplilified by PCR. Then the PCR product was cloned into pTZ57R/T cloning vector, The PT-LACK recombinant plasmid was extracted from transformed E.coli bacteria [TG1 strain] and sequenced. The LACK gene [Accession no LmjF28.2740] of MRHO/IR/75/ER and L. major was amplified using PCR method. LACK gene was cloned into pTZ57R/T coloning vector. Sequence analysis of the cloned LACK gene showed high homology 89% with LmjF28.2740 [LACK gene]. The LACK gene of L.major was cloned in pTZ57R/T vector successfully. Recombinant plasmid was confirmed and could be used for the production of recomiomort antigens is DNA vaccines, for further studies