Association of immune responses to mycobacterium tuberculosis peptide antigens with host genetic factors

Apolymerase chain reaction [PCR], based on insertion sequence IS6110,was developed to detect mycobacterium tuberculosis complex organisms in the blood samples of 56 tuberculosis patients and 34 healthy controls.the erly secreted antigenic target 6-KDa[ESAT-6] are used to stimulate T lymphocyte subsets from tuberculosis infected patients and the correltion of these immune responses to the genetic factoes[HLATYPE] which determined the host immune response is evaluated.ESAT-6 derived peptides:P1[1.05 +/- 0.084],P2[1.08 +/- 0.094],P3[1.02 +/- 0.086],P5[0.98 +/- 0.117] and P7[1.26 +/- 0.152] were significantly higer in the infected group than in non-infected one. Besides, 33ptients and 12 controls were tested for HL-DRB HLA DQB and HLA-DPB Only type HLA-DEB 15was significan tly associated with tuberculosis infection using the Chi-square test[X[2]=0.04311]. By using the relative risk some HL types were relatively more susceptible to be associated with tuberculosis infection. HLA-DR typing of patients showed that they covered a large spectrum of HLA-DR molecules encoded by HLA-DEB1,-DRB3,-DRB4,and DRB5genes.however,HLA-DQ typing showed that they HLA-DQB1 molecules, HLA-DP typing of patients showed that covered a large spectrum of HLA-DP molecules encoded by HLA-DPB1.

Production of antigenic recombinant cholera toxin subunit B.

Cholera is a major public health problem confronting developing countries, where outbreaks occur in a regular seasonal pattern and are particularly associated with poverty and poor sanitation. Primer set was designed to flank binding region of cholera toxin subunit B [ctxB] gene that was amplified using PCR. PCR products were purified using gel purification technique. Cholera ctxB gene was cloned into cloning vector. Stop codons contained in the insert were deleted and then the insert was subcloned into pQE expression vector. Cloned vectors were subjected to DNA sequence analysis. Small-scale culture was done for preparative rCTB production and purification. Mice were immunized with purified rCTB to test ability of rCTB to elicit antibody production. Primer set showed the ability to detect and excise ctxb gene successfully form cholera genomic DNA by PCR. After deleting the stop codons, sequence analysis revealed that the insert was in open reading frame with start codon of the vector. Small bacterial cultures revealed the presence of specific band at approximately 12-125 Kda in induced culture. rCTB was able to elicit specific antibody production after animal immunization

Antigenic potential of whole cell-recombinant cholera toxin subunit B [WC-CTB] preparation

Colonisation of the small intestine by V. cholerae, a typical non-invasive pathogen, is an important early step in the pathogenesis of cholera. In the present study, trial to make cholera whole-cell vaccine with addition of recombinant B subunit of cholera toxin [rCTB] was-done. Cholera chB gene was cloned into pQE expression vector. Large-scale culture was done for preparative rCTB production and purification. Commercial CTB from V. cholerae was used for induction of polyclonal anti-CTB antibodies in mice. These polyclonal antibodies were used to test the antigenicity and identity of rCTB. Cholera whole-cell vaccine was prepared by resuspending equal volumes of dead Inaba and Ogawa vaccine strains in PBS. Orochol E Berna was used as control vaccine. Mice were divided into 4 groups [20 mice each]; group 1 [G1]: control unimmunised, group 2 [62]: rCTB immunized, group 3 [G3]: rCTB + killed Inaba and Ogawa immunised, and group 4 [G4]: Orochol immunized. Sera and faeces from all groups were collected and used in evaluation of anti-rCTB antibody levels. The spleens of animals were aseptically removed and used in lymphoproliferative assay. Small bacterial cultures revealed the presence of specific band at approximately 12-12.5 KDa in induced culture. Anti-commercial CTB antibodies were successfully prepared and used in Western blot analysis and verified the presence and antigenicity of rCTB. Large scale production and purification of rCTB resulted in 12.9 mg protein. Both serum and secretory antibody level in mice of G2 was significantly less than in mice of both G3 and G4. G3 was significantly higher than 62, while there was no significant difference between G3 and G4. G4 was significantly higher than G2, while there was no significant difference between G4 and G3. Stimulation index of splenic cells in G 1 was not significantly different from that of G2, while it was significantly lower than GB and G4. SI of G2 was significantly lower than G3 and G4. G3 was significantly higher than G1, G2, and G4. G4 was significantly higher than 01 and 02, but it was significantly lower than G3. The results in this study showed the ability of rCT B to induce immune responses in the presence of cholera bacteria more than if used alone. The inclusion of rCTB in addition to vibrios, increased systemic, intestinal, and cellular responses. Depending on previous studies, adding any new cholera strain which are present or will appear in future is possible. In addition, other vaccines [either killed bacteria or vaccine subunit] can be added to the formula indicated in this study

Melatonin, the hormone of the pineal body: its role as an endogenous biological marker in major depression and breast cancer

The present study was designed to assess the relation of melatonin to two important pathological conditions namely; major depression [MD] and breast cancer [BC], with the intention of evaluating its role as an endogenous biological marker of both conditions and its potential clinical significance in follow-up of such cases. For this purposes 50 female patients with major depression [20 patients before treatment and 30 patients under treatment] in addition to 73 female patients with breast cancer [28 in stages I and II : early BC; 25 in stages III and IV late BC and 20 after radical mastectomy] were chosen for assessment of the serum melatonin levels. Nocturnal blood samples were collected from the MD group. whereas morning samples were collected from the BC group. Results were compared to those of an age-matched control group consisting of 20 healthy females. Nocturnal serum melatonin levels were significantly decreased in MD patients before the start of therapy as compared to the control group [P < 0.0001]. Meanwhile, the results of patients under antidepressant therapy [monoamine oxidase inhibitors and tricyclic antidepressants] showed no significant difference from the control group [P>0.05]. In cases of cancer breast, morning serum melatonin levels were significantly decreased in the early stages of the disease [P<0.0001], but became significantly elevated with progress of cancer and the occurrence of metastasis [P<0.0001]. Following radical mastectomy, the level of melatonin was insignificantly different from the control group [P >0.05]. Hence, we can conclude that decreased nocturnal melatonin could be considered an endogenous marker of major depressive illness, with such a decrease being masked by antidepressant therapy. Meanwhile, morning melatonin levels are of value in assessment and staging of breast cancer as well as post-operative follow-up of these patients. hopefully, aiming at early detection of recurrence

Effect of novalgin on chromosomes of rattus norvegicus

Novalgin; the sodium sulphonate derivative of aminopyrine, is an analgesic, antipyretic drug, widely used in Egypt. In spite of its well known hematologic complications, mutagenicity tests are deficient in the literature. The main aim of the present work was to study the cytogenetic effects of novalgin on bone marrow cells of Rattus norvegicus, using the method recommended by the Ad Hoc Committee of Mutagenicity Testing [1972]; Novalgin induced significant Chromosomal aberrations of both chromatid and chromosome type, and depression of mitosis 48 hours after treatment with the maximum tolerated dose in the acute study and after sub-acute treatment with the clinical dose, The results of the study suggest that novalgin is mutagenic and causes depression of mitosis when administered at very high doses, or after prolonged use of the clinical dose