Regulatory role of T Helper 17 (TH17) cells in autoimmune diabetes

Thl7 cells are thought to be involved in various autoimmune diseases such as multiple sclerosis, rheumatoid arthritis and experimental autoimmune encephalomyelitis. The role of Thl7 cells in the pathogenesis of autoimmune or type I diabetes (TID) was thus investigated using the non-obese diabetic (NOD) and the non-obese diabetic resistant (NOR) control mouse models. The expression of various cytokines in the Thl7 cells was assessed using flow cytometry following incubation with the peroxisome proliferator activated receptor gamma (PPARy) agonists, ciglitazone and 15-prostagalndin J2, and the PPARy antagonist, GW9662 to further delineate the influence of this immunoregulatory nuclear receptor on the Th 17 cell function. Basically, the constitutive level of the signature IL17A expression by Thl7 cells in NOD mice was more than that of NOR mice and that ciglitazone enhanced the expression of this cytokine in NOD mice whereas 15-PGJ2 and GW9662 had no effect. Interestingly, despite eliminating IFNy-expressing cells (Thl) at the start of culture, Th17 cells of NOD mice expressed high levels of this cytokine upon culture with ciglitazone suggesting that Thl7 may convert to Thl-like cells in TID in the presence of this PPARy agonist and may contribute to the pathogenesis of this autoimmune diseas

Gene quantitation by a competitive quantitative PCR technique using a homologous standard

The project aimed to develop a quantitative PCR technique to accurately quantity gene expression. We tested the accuracy of the method by determining the expression levels of the transcription factor, PPARy. This was done firstly in a mock system (utilizing cloned PPARy in a plasmid with known concentrations) and later on activated monocytes. The method utilized a homologous internal standard in a competitive PCR. This method has the advantage over using housekeeping genes as control since the latter requires different primers and amplification conditions. The current method also has advantage over using PCR MIMIC which may have different amplification efficiency due to the different gene fragment used. The homologous standard was constructed from the same target gene with a slightly shorter length to allow for visualization and analysis following competition with the target gene. The mock experiment showed that the method worked successfully. Using cDNA prepared from activated monocytes we were also able to show that it is useful for determining changes in the levels of PPARy expression in cells expressing the transcription factor

Lymphocyte activation status in HIV infected intravenous drug users (IVDU) in Kelantan

Infection with HIV results in profound immunologic changes in the infected host. The host immune system becomes dysregulated and profoundly suppressed culminating in a condition known as AIDS. Although this concept is well accepted, the actual immunopathology of the disease is far from established.Accumulated evidence have shown that the activation or misactivation of the immune system itself may contribute to disease progression (Pantaleo and Fauci, 1994).Thus, higher levels of activated CDS+T cells have been shown to correlate with poorer clinical status and lower CD4+T cell count (Giorgi et al., 1993). Although measurement of the CD4+ T cell level is the current "gold standard" used as a prognostic marker of HIV-disease, it is highly variable among different individuals or even within the same individual.In fact,we have recently demonstrated that the percentage of C04+T cell, a marker suggested to be more stable and less variable than the CD4+T cell absolute count (Taylor et al., 1989) may not necessarily be suitable to be used as a HIV-disease marker for our local stituation (Norazmi and Suam,1994). These data point to the need for separate studies in populations and risk groups other than white homosexual men to identify and clarify prognostic indicators for the development of AIDS

The use of assembly PCR for cloning of a malarial epitope into mycobacterium smegmatis

Mycobacterium bovis bacille Calmette Geurin (BCG) has been suggested to be an attractive vehicle for the delivery of various vaccine candidates,including those for malaria. However,despite possessing strong immunoadjuvant properties, recombinant BCG containing these malarial epitopes fail to elicit significant specific immunogenicity in mice.One possibility for this may be the codon bias between these organisms: mycobacteria is G:C rich while plasmodia is A:T rich. To test this hypothesis, we cloned the candidate malarial epitope, the C-terminus of Plasmodium falciparum merozoite surface protein-1(MSP-lC) using assembly PCR into Mycobacterium smegmatis. This technique involved the generation of the full 300bp fragment from 18, overlapping 40bp oligonucleotides designed in favour of mycobacteria codon usage. The fragment was cloned into the M.leprae l8kDa gene driven by the mycobacterial hsp60 promoter in a plasmid designated pUS937. The whole hsp60/18kDa/MSP-1C cassette was then cloned into a mycobacteriai replicative plasmid, pUS972, to produce pUS1754. A clone (pUS1758) containing the 'native' homologue of MSP-lC generated by PCR of genomic P. falciparum DNA was also constructed. Western blot analyses using an antibody against the 18kDa epitope showed that recombinants containing pUS 1754 had approximately 100-fold higher expression levels than those containing pUS1758. Two additional clones containing the 'native' MSP-lC also showed low level expression as compared to the clone containing pUS1754. Thus overcoming codon bias for selected vaccine candidates may be important in increasing the level of their expression and hence could improve the immunogenecity of such epitopes cloned into mycobacteria

Gene frequencies of D16S539, D7S820 and D13S317 STRs alleles in random malay population

DNA analysis of biological materials for individualization purposes has become the norm In crime investigation. A number of different DNA markers exist for individualization of biological stains. However STR based DNA profiling(microsatellite polymorphism) is the most widely used technique in personal identification. Personal identification tests involve mathematical probabilities of genetic markers to ascertain the likelihood of probability for a conclusive answer. In this study distribution of allele frequencies of three STR polymorphic genetic markers-viz .,D13S317,D7S820 and 0168539 were studied in random malay population of Malaysia. The observed genotypic distribution showed no significant deviation from Hardy-weinberg equilibrium . The data in the present study have been compared with recently published data as reported in the liuterature for various population groups. The data on the three validated STRs will be useful in parentage testing and personal identification in criminal and immigration cases

Immunofenotype mononucleus cell peripheral malaria patients

The absolute number and percentage of various T cell subsets in patients with acute malaria infection was investigate with special reference to the expression of the T activation markers, CD25,HLA-DR and CD38. The absolute number but not the percentage of CD4+(p<0.001) and CD8 (p<0.02) T cell subsets as well as total B cells (CD19+) (p<0.02) was reduced in these patients. In contrast, the percentage of T cells bearing the gamma-delta (gd) T-cell receptor (TCR) was significantly increased (p<0.02). Although the expression of CD25 on any of the T cell subsets with the normal range, increases in the percentage of CD8+CD38 + (p<0.005) gd+HLA-DR+ (p<0.001) and gd+CD38+ (p<0.005) T cell subsets may be suggestive inter-relationship with the disease the significant of which remains to of their be ascertained

Antibodi monoklon terhadap sel mononukleus darah periferi manusia

Dalam kajian ini, teknik hibridoma talah dilakukan untuk menghasilkan antibodi monoklon (mAb) mencit terhadap sel mononukleus darah periferi manusia( SMDP). Selepas hibridisasi sel limpa mencit yang diimunisasi dengan sel mieloma dilakukan, sel hibridorna yang diperolehi didapati stabil dan mengeluarkan antibodi(Ab)yang berterusan.Dua hibridoma telah berjaya dihasilkan daripada pengklonan satu kultur awal berdasarkan analisis blot imunodot. Kedua-dua hibridoma ini menghasilkan Ab yang masing-masingnya dikenali sebagai PPSP1 dan PPSP2. Analisis selanjutnya melalui kaedah flow sitometri menunjukkan bahawa kedua-dua hibridoma ini menghasilkan Ab yang bertindak balas dengan antigen permukaan SMDP. Walau bagaimanapun, intensiti pewarnaan PPSP1 lebih tinggi daripada PPSP2. Lebih daripada 98% SMDP label positif dengan PPSP1 dan PPSP2 iaitu setanding dengan keputusan apabila menggunakan mAb kontrol positif komersial CD45.Kedua-dua AB ini besar kemungkinan adalah mAb. Bagaimanapun, pencirian selanjutnya perlu mengenalpasti kespesifikan PPSP1 dan PPSP2

Special elements and values needed in leadership for special education

Educational leadership is very important in determining the success of an educational organization, especially in schools. Effective leadership is more needed when it involves more challenging and diverse situations such as in special education. Many competent school leaders have succeeded in making the school organizations they lead as successful. Nevertheless, the same leaders are seen to be less effective on leadership in special education. Therefore, this study was conducted to explore the elements and added value that a leader must have to lead special education effectively. This qualitative study was conducted by involving an interview session with 20 special education teachers in the state of Johor, Malaysia. Their perceptions and opinions are analyzed to obtain the relevant elements and added value. As a result of the thematic analysis conducted, there are five additional elements that need to be practiced by special education leaders in schools in each situation, namely loving, caring, sensitive, friendly and cooperative network. These five elements were agreed upon by the respondents involved and should be given attention in every decision made by leaders in special education. It is hoped that the findings of this study can help special education leaders in schools to practice effective leadership and in turn lead the continued success of special education

Immune deficit in recurrent pyogenic cholangitis

As the name suggests, patients with recurrent pyogenic cholangitis (RPC) are prone to repeated episodes of cholangitis with systemic sepsis. The mechanism and pathogenesis of sepsis in these patients is not well understood, although some animal studies have suggest a functional defect in the immune system. To determine whether an immunological deficit may account for RPC, we conducted a pilot study to assess the immune status of these patients and compared it to a control group. Using two-colour flow cytometry, we found that the absolute number of various lymphocytes subpopulations, namely total T cells (CD3+), CD4+ T cells, total B cells (CD19t) and natural killer (NK) cells (CD16+56+) seemed to be depressed in patients with RPC as compared to the control group. In addition we observed that these patients had higher levels of CDS+ cells expressing the CD38 activation marker. Although based on a small sample size, these findings suggest that immune depression may be an important feature of RPC