Quantitative Reference Ranges for Fasting Profiles and Oral Glucose Tolerance Test for Healthy Adults in Metropolitan Region of Nairobi; Kenya

Purpose:To establish quantitative reference ranges for fasting profiles and oral glucose tolerance test for healthy adults in metropolitan region of Nairobi. Methods: A prospective study carried out on 871 healthy subjects from the metropolitan region of Kenya. Results: The fasting profile parameters investigated were fasting blood glucose (FBG); total cholesterol (TC) triglycerides (TG); high density lipoprotein cholesterol (HDLC); low density lipoprotein cholesterol (LDLC) and TC/HDLC ratio. In addition; oral glucose tolerance test (OGTT) was also investigated. Eight hundred and seventy one (871) healthy study subjects were involved in the study. Established reference ranges were as follows: FBG (venous whole blood) (2.1 - 5.7) mmol/L; TC (2.9 - 6.4) mmol/L; TG (0.44- 2.44); HDL C (1.1 - 2.1) mmol/L; LDLC (1.1 - 4.3) mmol/L; TC/HDLC ratio (1.1 - 5.4). Established reference ranges for oral glucose tolerance test (OGTT) were as follows: baseline/fasting blood glucose capillary whole blood (3.2-5.4) mmol/L; half hour (4.7-8.9) mmol/L; one hour (4.4-9.8) mmol/L; one hour and half (4-8.1) mmol/L and two hours (3.4-7.2) mmol/L. Results for gender differences for the studied parameters were as follows: FBG (p=0.124); TC (p=0.205); TG (p=0.705) HDLC (p= 0.52); LDLC (p=0.417) and TC/HDLC ratio (p=0.359). On the other hand; the gender results for timed OGTT were as follows: 0 hour (p=0.123); half hour (p=0.479); one hour (p=0.412); one hour and half (p=0.596)) and two hours (p=0.630). Hence there were no gender disparities for the parameters in the studied adult Kenyan population. Conclusion: Since the established reference ranges are a reflection of the Kenyan adult population our clinical chemistry laboratory reports interpretations will henceforth be independent of what has been quoted in literature. Likewise effective diagnosis and management of glucose and lipids pathological disorders will be achieved by the use of established adult Kenyan reference ranges

Subclinical nephrotoxicity associated with occupational silica exposure among male Kenyan industrial workers.

OBJECTIVE: To determine early signs of renal injury due to occupational silica exposure. DESIGN: Cross-sectional analytical research. SETTINGS: Kenyatta National Hospital for the referent population and Clayworks ceramics, bricks and tiles factory for the assessment of occupational silica exposure. SUBJECTS: Thirty three non-smoking silica-exposed male industrial workers and 38 non-smoking male referents participated in this study. RESULTS: Silica-exposed males excreted significantly increased levels of U.TP, U.Malb, U.ALP, U.y-GT and U.LDH compared to referent males. Among the silica-exposed males, U.Si negatively correlated significantly with age, U.TP correlated significantly to each of U.ALP and U.LDH. However, no correlation was observed between work duration and U.Si. CONCLUSION: The present study shows that there is associated glomerular and proximal tubular damage among silica exposed workers which is not duration related and is seemingly subclinical and nonprogressive and urinary silica levels appears to be similar in all groups and are not affected by exposure and work duration: the reason for which is unclear.

A subset of Cowdria ruminantium genes important for immune recognition and protection.

Cowdria ruminantium causes the tick-borne rickettsial disease of heartwater, which is devastating to livestock production in sub-Saharan Africa. Current diagnosis and control methods are inadequate. We have identified and sequenced a subset of genes encoding recombinant antigens recognized by antibody and peripheral blood mononuclear cells from immune ruminants. The identified genes include many with significant similarity to those of Rickettsia prowazekii, genes predicted to encode different outer membrane proteins and lipoproteins and a gene containing an unusual tandem repeat structure. Evidence is presented for immune protection by recombinant antigens in a mouse model of C. ruminantium infection. These data identify new recombinant antigens for evaluation in vaccines and diagnostic tests to control heartwater.

Monoclonal antibody binding to a surface-exposed epitope on Cowdria ruminantium that is conserved among eight strains.

Monoclonal antibodies (MAb) binding to Cowdria ruminantium elementary bodies (EB) were identified by enzyme-linked immunosorbent assay, and surface binding of one MAb (446.15) to intact EB was determined by immunofluorescence, immunogold labeling, and transmission electron microscopy. MAb 446.15 bound an antigen of approximately 43 kDa in immunoblots of eight geographically distinct strains. The MAb did not react with Ehrlichia canis antigens or uninfected bovine endothelial cell lysate and may be useful in diagnostic assays and vaccine development.

A highly sensitive, non-radioactive assay for T cell activation in cattle: applications in screening for antigens recognised by CD4(+) and CD8(+) T cells.

We describe a highly sensitive, non-radioactive assay for T cell activation, based on the rapid induction of class II MHC expression by constitutively negative bovine endothelial cells, when cultured in the presence of supernatants derived from activated bovine T cells. We demonstrate the effectiveness of this assay in detecting rBoIFNgamma and activation of immune CD4(+) and CD8(+) T cell lines and clones in response to specific antigen and transfected COS-7 cells, respectively. We also demonstrate its utility in identifying purified pathogen fractions that activate immune CD4(+) T cell clones.

Comparative parasitological and haematological changes in two breeds of sheep infected with Fasciola gigantica.

Twelve each of Red Masai and Dorper sheep, aged between 6 and 9 months, were acquired from a Fasciola-free area of eastern Kenya. Each breed was divided into two groups of 6. The sheep in one group of each breed were experimentally infected with 400 viable metacercariae of Fasciola gigantica. The other group of 6 sheep of each breed remained as uninfected controls. The animals were monitored regularly for any evidence of disease. Blood samples taken weekly revealed a general reduction in red cell counts and packed cell volume, which was much faster in the infected Dorper sheep than in the Red Masai. This reduction started from the tenth week after infection and persisted to the end of the experiment 18 weeks post infection (PI). The absolute eosinophil counts rose in all the infected animals, but the values were higher among the Dorper than among the Red Masai. Patency occurred at weeks 12 and 13 PI in the Red Masai and Dorpers, respectively, with the latter shedding significantly more fluke eggs. The worm recovery rates were higher among the Dorpers than among the Red Masai, though not significantly so. On the basis of egg counts and clinicopathology, the Dorper sheep were considered to be more susceptible to F. gigantica infections.

A comparison of serum biochemical changes in two breeds of sheep (Red Masai and Dorper) experimentally infected with Fasciola gigantica.

Twelve Red Masai and 12 Dorper sheep aged between 6 and 9 months, were acquired from a fluke-free area and sheep of each breed divided into two equal groups of six. Each animal in one group of each breed was experimentally infected with 400 viable metacercariae of Fasciola gigantica. The other groups acted as uninfected controls. Blood samples were taken at weekly intervals for the determination of serum bilirubin, albumin, and gamma glutamyl transferase levels. Following the establishment of infection, albumin levels declined in both breeds of infected animals without any significant difference between the two breeds. However, serum bilirubin and gamma glutamyl transferase (GGT) in the infected animals were elevated significantly more in the Dorper than in the Red Masai sheep. Based on these findings, it would appear that Dorper sheep are more susceptible to the infection than Red Masai sheep.

Immunization of cattle by infection with Cowdria ruminantium elicits T lymphocytes that recognize autologous, infected endothelial cells and monocytes.

Peripheral blood mononuclear cells (PBMC) from immune cattle proliferate in the presence of autologous Cowdria ruminantium-infected endothelial cells and monocytes. Endothelial cells required treatment with T-cell growth factors to induce class II major histocompatibility complex expression prior to infection and use as stimulators. Proliferative responses to both infected autologous endothelial cells and monocytes were characterized by expansion of a mixture of CD4+, CD8+, and gammadelta T cells. However, gammadelta T cells dominated following several restimulations. Reverse transcription-PCR analysis of cytokine expression by C. ruminantium-specific T-cell lines and immune PBMC revealed weak interleukin-2 (IL-2), IL-4, and gamma interferon (IFN-gamma) transcripts at 3 to 24 h after stimulation. Strong expression of IFN-gamma, tumor necrosis factor alpha (TNF-alpha), TNF-beta, and IL-2 receptor alpha-chain mRNA was detected in T-cell lines 48 h after antigen stimulation. Supernatants from these T-cell cultures contained IFN-gamma protein. Our findings suggest that in immune cattle a C. ruminantium-specific T-cell response is induced and that infected endothelial cells and monocytes may present C. ruminantium antigens to specific T lymphocytes in vivo during infection and thereby play a role in induction of protective immune responses to the pathogen.

Cellular immune responses of cattle to Cowdria ruminantium.

Peripheral blood mononuclear cells (PBM) from cattle immunised against Cowdria ruminantium infection (Heartwater), proliferated in vitro in the presence of either infected autologous endothelial cells pre-treated with T cell growth factors to induce MHC class II expression, or infected autologous monocytes. Proliferation was not observed in PBM cultured with a soluble extract of the agent, but PBM responded to two recombinant antigens of C. ruminantium, namely a 32 kDa (MAP1) and a 21 kDa antigen (MAP2). We hypothesize that infected endothelial cells and monocytes present Cowdria antigens to specific lymphocytes during infection and thereby play a role in the pathogenesis/immune response to the pathogen.

The effects of a controlled-release albendazole capsule (Proftril-Captec) on parasitism in grazing Corriedale ewes in the Nyandarua district of Kenya.

The effects of intraruminal sustained-release capsules (IRSRCs) on parasitism in grazing Corriedale ewes were investigated over a period of 119 days (4 June-30 September 1993) using 40 ewes aged approximately 2 years and randomly divided into two groups of 20 ewes each. Each of the ewes in the treatment group received an IRSRC while the controls were left untreated. The groups were placed on adjacent 2.5-acre paddocks obtained by subdividing a 5.0-acre permanent pasture which had previously been grazed by young untreated sheep, so exposing both groups of ewes to a similar challenge from a contaminated paddock. The faecal egg counts, herbage larval counts and worm burdens of the major gastrointestinal parasites of sheep were significantly reduced by the use of the IRSRC. These parasitological effects were reflected in the increased live weight gains and heavier fleeces of the IRSRC-treated ewes. The control ewes required occasional salvage treatments during the trial period and the herbage on their paddock was heavily contaminated with infected larvae, reflected in the high worm burdens in the control ewes necropsied at the end of the trial and in tracer sheep introduced into the paddocks during the initial (day 30), interim (day 58) and final (day 89) stages of the experiment. Most capsules were exhausted within 95 days of administration, leading to a rise in the count of eggs per gram in the faeces in the treated group towards the end of the study.

Trypanosoma congolense infection in sheep: cellular phenotypes in lymph and lymph nodes associated with skin reactions.

Intradermal inoculation of sheep with culture-derived metacyclic forms of Trypanosoma congolense resulted in the development of localized skin reactions (chancres) and enlargement of the draining lymph nodes 7 days after infection. Changes in the expression of surface antigens of lymphocytes in lymph leaving the affected skin reactions and in the associated lymph nodes were monitored by cannulating the afferent and efferent lymphatic ducts. Trypanosomes appeared in afferent and efferent lymph 3 to 5 days after infection and persisted even as the chancres regressed. The cellular output in both afferent and efferent lymph increased markedly after the onset of parasitosis. Sequential analysis of the phenotypes of lymphocytes by immunofluorescent staining and flow cytometry revealed that in afferent lymph draining the chancre there was an early response which was due to an increase in T cells, particularly CD4+ and CD8+ cells; however, as the chancres-regressed there was an increase in lymphoblasts and surface immunoglobulin-bearing cells. In contrast, in the efferent lymph, the increase in lymphocytes was due predominantly to a higher number of cells bearing surface immunoglobulins.

Trypanosoma congolense infection in sheep: ultrastructural changes in the skin prior to development of local skin reactions.

Events occurring in the skin of sheep prior to development of Trypanosoma congolense-induced local skin reactions (chancres) were studied using electron microscopy. Three days after infection, few trypanosomes were present in the dermal collagen. However, these parasites were more abundant 5 days after infection, and were also found in dermal lymphatics and in the connective tissue matrix between collagen bundles. Mast cells in the skin obtained 5 days after infection showed evidence of degranulation. These events may play a role during the induction phase of trypanosomal chancres.

Immunohistology of lymph nodes draining local skin reactions (chancres) in sheep infected with Trypanosoma congolense.

Marked enlargement of lymph nodes draining local skin reactions (chancres) occurred in sheep following intradermal inoculation of cultured metacyclic forms of Trypanosoma congolense. Histologically, these lymph nodes were characterized by follicular hypertrophy and hyperplasia, compression and relative reduction of the paracortical areas and expansion of the medullary regions. Immunohistochemical staining with monoclonal antibodies to ovine lymphocyte subsets and Fc receptor (FcR) bearing macrophages, revealed increased expression of B cells (CD45R+), major histocompatibility complex (MHC) Class II, FcR+ macrophages, and CD1+ cells in the cortical and paracortical areas. The paracortical areas were found to be sparsely populated by CD5+, CD4+ and CD8+ cells, while the medullary areas contained numerous CD8+ cells and FcR+ macrophages. FcR+ macrophages were also present in cortical trabecular and subcapsular sinuses. As the chancre regressed, lymph node reactivity also subsided and fewer B cell follicles were observed and there was decreased expression of CD45R+ and MHC Class II+ cells.

Cellular phenotypes in Trypanosoma congolense infected sheep: the local skin reaction.

Mononuclear cell subpopulations in local skin reactions (chancres) in sheep infected with metacyclic forms of Trypanosoma congolense were studied by indirect immunoperoxidase staining using a panel of monoclonal antibodies (MoAbs) specific for ovine leucocyte subsets. Morphometric analysis revealed significant increases in numbers of cells expressing CD5, CD4, CD8, CD45R (mainly B cells), major histocompatibility complex (MHC) class II antigens, Fc receptors (FcR) on macrophages (VPM32) and FcR on B cells and macrophages (VPM33) from five days post-infection. B cells which also expressed MHC class II were found mainly in dense aggregates. The CD4/CD8 ratios were raised over pre-infection levels at 5-7 days post-infection. In sheep which had been infected, treated with trypanocidal drugs and then challenged with an heterologous serodeme of T. congolense, changes in cellular phenotype kinetics were similar to those seen in the skin in primary infections. Sheep superinfected with either an homologous or an heterologous, T. congolense serodeme showed only mild cellular infiltration and slight increases in various cellular phenotypes at the sites of inoculation.

Immunosuppression in caprine trypanosomiasis: effects of acute Trypanosoma congolense infection on antibody response to anthrax spore vaccine.

Trypanosoma congolense infected goats were vaccinated with Bacillus anthracis spore vaccine to determine the effect of such infection on the humoral immune response to the vaccine. The anti-anthrax antibody levels were severely depressed in infected goats. When trypanocidal therapy was administered to T. congolense infected goats 14 days after infection they developed antibody levels against Bacillus anthracis similar to uninfected controls.