Transferrin levels are associated with malnutrition markers in hemodialysis patients in KwaZulu-Natal, South Africa.

INTRODUCTION: Malnutrition is a global phenomenon and may be contributing to the increasing size of the hemodialysis (HD) population in South Africa and is affecting morbidity and clinical outcomes. Our study assessed whether transferrin could be a possible marker for malnutrition in the HD population. METHODS: Clinical parameters (including skinfold thickness and mid-upper arm circumference [MUAC]) and laboratory markers (including transferrin and hemoglobin) were measured during a six-month period in a sample of 59 HD patients. RESULTS: Linear regression analysis showed that MUAC (p = 0.027) as well as skinfold thickness (p = 0.021) had a significant association with transferrin levels within the HD participants. There was no significant association between transferrin levels or MUAC with hemoglobin levels (p = 0.075). Furthermore, the study found that decreased transferrin levels (< 2.15 g/dL to 3.80 g/dL) were closely related to malnutrition in the malnutrition distribution groups within the study, with 97.7% of HD participants being classified in one of the malnutrition groups. CONCLUSION: Thus, transferrin levels are a valuable marker for malnutrition within the HD patient population and can be included along with clinical assessment parameters such as MUAC and skinfold thickness as primary indicators for malnutrition.

Regulatory T cell frequency in peripheral blood of women with advanced cervical Cancer including women living with HIV.

BACKGROUND: Persistent high-risk Human papillomavirus (HR-HPV) infections are the main cause of cervical cancer. Cumulative evidence implicates regulatory T cells (Tregs) as a critical factor in the failure to eliminate HPV-induced cancers leading to their persistence and progression to cancer. Also, the WHO recognised cervical cancer as 100% attributable to persistent HR-HPV infection. The province of KwaZulu-Natal (KZN) in South Africa has a high prevalence of cervical cancer and HIV infection. MATERIALS AND METHODS: We evaluated Treg frequency in dual infection of HR HPV and HIV coinfection using phenotypic markers, CD4, CD25 and intracellular Foxp3, in the peripheral blood of 51 cervical cancer and 46 non-cervical cancer participants and evaluated the effect of HIV on regulatory T cell proportion. Peripheral blood mononuclear cells were surface stained with a cocktail fluorescent labelled CD4 and CD25 and subsequently with APC anti-human FoxP3 (eBioscience). Flow cytometry was performed with FACS analysis. Statistical analysis of results was done using Instat 3 program (GraphpadR). Tregs results were expressed as median ± interquartile range (IQR). Associations of cervical cancer with demographic, clinical and laboratory variables were evaluated by univariate and multivariate logistic regression analysis using SPSS version 27 (IBM). RESULTS: Tregs frequency was significantly higher in individuals with cervical cancer (11.00 ± 19.79%) compared to controls (1.71 ± 8.91%) (p < 0.0001). HIV infection was associated with an increase in Tregs frequency. In controls a significant difference in Tregs frequency was noted between women living with HIV (6.00 ± 10.57%, n = 9) and those without HIV (1.30 ± 6.10%, n = 37), p = 0.0023. In multivariate logistic regression, Tregs frequency was significantly associated with cervical cancer after controlling for age, smoking, weight loss, presence of STI, HIV and HPV genotype. DISCUSSION/CONCLUSION: Higher Tregs frequency was significantly associated with cervical cancer highlighting the immunosuppressive role of Tregs in cervical cancer. Treg frequency was more strongly associated with cervical cancer than HIV infection. We provide baseline data for monitoring Treg frequencies in response to new preventive and therapeutic strategies in the management of cervical cancer.

The prevalence of chronic kidney disease in South Africa - limitations of studies comparing prevalence with sub-Saharan Africa, Africa, and globally.

BACKGROUND: Chronic kidney disease (CKD) is a globally significant non-communicable disorder. CKD prevalence varies between countries and within a country. We compared the prevalence rates of CKD in South Africa with sub-Saharan Africa, Africa, and globally. METHODS: We registered a systematic review with the International Prospective Register of Systematic Reviews for prevalence studies reporting CKD stages III-V from 2013 to 2021. The analysis sought to explain any significant differences in prevalence rates. The R statistical package was used for data analysis. Comparisons included measures of effect size due to the large sample sizes analysed. We also compared sex differences in prevalence rates, common aetiologies, and type of study methodologies employed. RESULTS: Eight studies were analysed, with two from each region. The matched prevalence rates of CKD between the various regions and South Africa showed significant differences, except for one comparison between South Africa and an African study [p = 0.09 (95% CI - 0.04-0.01)]. Both sub-Saharan African studies had a higher prevalence than South Africa. One study in Africa had a higher prevalence, while the other had a lower prevalence, whilst one Global study had a higher prevalence, and the other had a lower prevalence compared to South Africa. The statistical differences analysed using the Cramer's V test were substantially less than 0.1. Thus, differences in comparisons were largely due to differences in sample sizes rather than actual differences. CONCLUSION: Variable prevalence rates between regions included disparities in sample size, definitions of CKD, lack of chronicity testing and heterogeneous laboratory estimations of eGFR. Improved consistency and enhanced methods for diagnosing and comparing CKD prevalence are essential.

Association of Staphylococcus nasal colonization and HIV in end-stage renal failure patients undergoing peritoneal dialysis.

INTRODUCTION: Staphylococcal infections can cause significant morbidity in patients undergoing dialysis. This study evaluated the effects of HIV infection on nasal carriage of Staphylococcus aureus, staphylococcal peritonitis, and catheter infection rates in patients with end-stage renal failure managed with continuous ambulatory peritoneal dialysis (CAPD). METHODS: Sixty HIV-positive and 59 HIV-negative CAPD patients were enrolled and followed up for up to 18 months. S. aureus nasal carriage (detected by nasal swab culture), Staphylococcal peritonitis (diagnosed by clinical presentation, and CAPD effluent Staphylococcal culture and white blood cell count ≥100 cells/µL), and catheter infections (including exit site and tunnel infections) were assessed monthly. RESULTS: At 18 months, S. aureus nasal carriage rates were 43.3% and 30.5% (p = 0.147) and the methicillin-resistant S. aureus (MRSA) nasal carriage rates were 31.7% and 13.6% (p = 0.018) for the HIV-positive and HIV-negative cohorts, respectively. The HIV-positive cohort was associated with increased hazards for staphylococcal peritonitis, (adjusted hazard ratio [AHR] 2.85, 95% confidence interval [CI] 1.19-6.84, p = 0.019) due to increased coagulase-negative staphylococcal (CNS) peritonitis rate in the HIV-positive cohort compared with the HIV-negative cohort (0.435 vs. 0.089 episodes/person-years; AHR 7.64, CI 2.18-26.82, p = 0.001). On multivariable analysis, CD4+ cell count <200 cells/µL, diabetes, and S. aureus nasal carriage were found to be independent predictors of S. aureus peritonitis. CONCLUSIONS: These findings suggest that HIV infection may be a risk factor for MRSA nasal colonization and may increase the risks of CNS peritonitis, while a CD4+ cell count <200 cells/µL and S. aureus nasal carriage may be important predictors of S. aureus peritonitis.

Detection of human immunodeficiency virus-1 ribonucleic acid in the peritoneal effluent of renal failure patients on highly active antiretroviral therapy.

Background: We evaluated the shedding of human immunodeficiency virus (HIV)-1 particles into continuous ambulatory peritoneal dialysis (CAPD) effluents of HIV-positive patients with end-stage renal disease (ESRD). Methods: A total of 58 HIV-positive patients with ESRD on highly active antiretroviral therapy (HAART) who had Tenckhoff catheters inserted between September 2012 and February 2015 were prospectively reviewed and followed for 18 months. Peritoneal dialysis (PD) effluent samples from functioning CAPD catheters and plasma samples were obtained at three points during regular clinic visits on days 45 ± 37, 200 ± 19 and 377 ± 13 after catheter insertion. All specimens were stored at -20°C, and each batch was analysed by Roche quantitative HIV-1 polymerase chain reaction assay to detect HIV-1 particles. Clustered logistic regression was used to test for independent predictors of HIV-1 detection in CAPD effluents. Results: HIV-1 RNA above 20 copies/mL assay limit was detectable in 19% (first batch), 26.3% (second batch) and 20% (third batch) of PD effluent specimens. HIV-1 RNA was detectable in PD fluid, without corresponding detection in the paired plasma in 3.4% (first batch), 5.3% (second batch) and 10% (third batch) of samples. Detection of HIV-1 in plasma sample (odds ratios 3.94; 95% confidence interval 1.14-13.55; P = 0.030), body mass index, serum albumin and HAART regimen were found to be significantly associated with HIV-1 detection in PD effluents. Conclusions: HIV particles are shed in detectable amounts into CAPD effluents even in patients with suppressed plasma viral load, raising concerns of a localized sanctuary site and potential infectivity of HIV-positive CAPD patients on a full complement of HAART.

Peritonitis outcomes in patients with HIV and end-stage renal failure on peritoneal dialysis: a prospective cohort study.

BACKGROUND: Few studies have investigated the management of human immunodeficiency virus (HIV)-associated end-stage renal failure particularly in low-resource settings with limited access to renal replacement therapy. We aimed to evaluate the effects of HIV infection on continuous ambulatory peritoneal dialysis (CAPD)-associated peritonitis outcomes and technique failure in highly active antiretroviral therapy (HAART)-treated HIV-positive CAPD populations. METHODS: We conducted a single-center prospective cohort study of consecutive incident CAPD patients recruited from two hospitals in Durban, South Africa from September 2012-February 2015. Seventy HIV-negative and 70 HIV-positive end-stage renal failure patients were followed monthly for 18 months at a central renal clinic. Primary outcomes of peritonitis and catheter failure were assessed for the first 18 months of CAPD therapy. We assessed risk factors for peritonitis and catheter failure using Cox regression survival analysis. RESULTS: The HIV-positive cohort had a significantly increased rate of peritonitis compared to the HIV-negative cohort (1.86 vs. 0.76 episodes/person-years, respectively; hazard ratio [HR], 2.41; 95% confidence interval [CI], 1.69-3.45, P < 0.001). When the baseline CD4 count was below 200 cells/µL, the peritonitis rate rose to 3.69 episodes/person-years (HR 4.54, 95% CI 2.35-8.76, P < 0.001), while a baseline CD4 count above 350 cells/µL was associated with a peritonitis rate of 1.60 episodes/person-years (HR 2.10, CI 1.39-3.15, P = 0.001). HIV was associated with increased hazards of peritonitis relapse (HR, 3.88; CI, 1.37-10.94; P = 0.010). Independent predictors associated with increased peritonitis risk were HIV (HR, 1.84; CI, 1.07-3.16; P = 0.027), diabetes (HR, 2.09; CI, 1.09-4.03; P = 0.027) and a baseline CD4 count < 200 cells/µL (HR, 3.28; CI, 1.42-7.61; P = 0.006). Catheter failure rates were 0.34 (HIV-positive cohort) and 0.24 (HIV-negative cohort) episodes/person-years (HR, 1.42; 95% CI, 0.73-2.73; P = 0.299). Peritonitis (HR, 14.47; CI, 2.79-75.00; P = 0.001), average hemoglobin concentrations (HR, 0.75; CI, 0.59-0.95; P = 0.016), and average serum C-reactive protein levels were independent predictors of catheter failure. CONCLUSIONS: HIV infection in end-stage renal disease patients managed by CAPD was associated with increased peritonitis risk; however, HIV infection did not increase the risk for CAPD catheter failure rate at 18 months.

Characteristics of South African patients presenting with kidney disease in rural KwaZulu-Natal: a cross sectional study.

BACKGROUND: Diabetes mellitus is the leading cause of end-stage renal disease (ESRD) globally. Diabetes and human immunodeficiency virus (HIV), both prevalent in South Africa, have not been reported as significant causes of ESRD. METHODS: We evaluated chronic kidney disease (CKD) and cardiovascular disease risk factors in a cross-sectional study of 302 patients (165 females/ 137 males) at a CKD clinic in rural northern KwaZulu-Natal. We included all CKD outpatient clinic attendees and excluded acute renal failure patients. Demographic, clinical and laboratory data collected were analyzed with Stata11 software. Logistic regression analysis was used to determine factors associated with advanced CKD and results expressed as the odds ratio with the 95% confidence interval [OR (95% CI)]. RESULTS: Of 302 patients analyzed, 290 (96%) were black African. Mean age ± SD was 47.1 ± 17.0 years. Approximately 86.4% of females and 54.5% of males were overweight/ obese. Dyslipidaemia was observed in 47.9% females and 29.2% males (P < 0.001). Estimated glomerular filtration rate (eGFR) was <30 ml/min/1.73 m2 in 50.6% patients. CKD risk factors observed were: hypertension (77.8%), diabetes (29.8%), HIV (28.5%), glomerulonephritis (7.0%) and tubulointerstitial diseases (5.6%). Independent factors associated with eGFR <30 ml/min/1.73 m2 at presentation were: HIV [OR = 2.4 (1.3-4.2), P = 0.004] and hypertension [OR = 2.3 (1.3-4.2), P = 0.007]. CONCLUSION: Diabetes and HIV are prevalent in CKD patients at primary/regional level healthcare in South Africa. With registry data lacking, dedicated CKD clinics at lower healthcare levels may provide valuable data on CKD epidemiology including changes in aetiology. Primary healthcare practitioners are faced with advanced CKD patients in resource-poor settings, with limited opportunity for upward referral hence the need for nephrology outreach programs.

Paradoxical Perforating Folliculitis in a Newly Transplanted Kidney Recipient: Case Study.

We report on a rare case of perforating folliculitis with a paradoxical presentation. An 18-year-old patient with end-stage kidney failure was undergoing continuous ambulatory peritoneal dialysis following 1 year of hemodialysis treatment. While being treated with continuous ambulatory peritoneal dialysis, he developed an itchy papular eruption on an erythematous base over his face and chest, which was diagnosed as chicken pox and treatedwith acyclovir.He also underwent successful deceased donor kidney transplant 1 year later. On day 10 posttransplant, he presented with a papular eruption over the chest, face, and forearms. A skin biopsy revealed a perforating folliculitis lesion. He was treated with prednisone and tacrolimus, as part of the kidney transplant treatment. The skin lesions resolved progressively. His urea, creatinine, and electrolyte levels remained normal and on an ever-improving trend at each visit. By 4 months posttransplant, the skin lesions had resolved almost completely.

Beta 2 microglobulin in kidney failure: A review and an algorithm for renal replacement therapy.

For decades, beta 2 microglobulin (B2M) has been a subject of great interest in nephrology and other fields such as multiple myeloma. B2M, a 99 amino acid protein, is associated with amyloid deposits in patients undergoing renal replacement therapy (RRT). The source of information is published articles on B2M in chronic renal failure since 1960. We have reviewed literature published since 1960 to date, highlighting the milestones of the role of B2M in chronic kidney disease (CKD) and B2M serum values in patients treated by various RRTs. B2M deposits associated with the disease include carpal tunnel syndrome, spondyloarthropathy, and arthritis of large joints such as the shoulders. The role of RRT in the removal of B2M in CKD is discussed. Recent reports include factors affecting the process of fibrillation and deposition of B2M in tissues. A comparative report of various modalities of treatment on the serum levels of B2M is provided. The presence of significant residual urine output in continuous ambulatory peritoneal dialysis patients may explain why peritoneal dialysis is a modality that is associated with the lowest level of serum B2M. Patients treated with hemodiafiltration or hemodialysis (HD) using high flux dialyzers have lower levels of B2M than those treated by HD with low flux dialyzers. Finally, based on the literature review, an algorithm for RRT using B2M level monitoring and other variables is proposed and needs evaluation in a controlled trial.

Antibodies to Erythropoietin Are Associated with Erythropoietin Resistance in Hemodialysis Patients in KwaZulu-Natal (South Africa).

Recombinant human erythropoietin (rHuEPO) is a glycoprotein and biological equivalent to the endogenous compound administered to treat anemia of end-stage renal disease patients. Resistance to rHuEPO has been reported, whereby patients require higher and higher doses of rHuEPO to maintain an adequate hemoglobin level. In this study, assessment of native and administered erythropoietin (EPO), antibody and hemoglobin levels was carried out on a sample of patients with renal failure on hemodialysis (HD). This is a randomized controlled trial where consecutive subjects attending HD units at Addington Hospital and King Edward Hospital, Durban (South Africa) were included until the target number was reached. Forty patients with renal failure on HD and receiving recombinant EPO Beta (Recormon) for treatment of anemia via the subcutaneous route in weekly doses of 2000 IU, 4000 IU, 6000 IU, 8000 IU, 12,000 IU, or 18,000 IU according to the severity of the anemia were included after obtaining informed consent. Also included in the study were 10 HD patients not on rHuEPO therapy and 10 healthy individuals from the Durban University of Technology, recruited as described above to form the control group. ELISA was used to measure serum levels of EPO as well as antibodies to EPO. Results were analyzed by descriptive, inferential methods and by logistic regression analysis using IBM SPSS Statistics for Windows version 22.0. Antibodies to EPO were found in almost all patients who were receiving EPO. The highest levels of antibody to EPO were found to be associated with patients receiving the highest weekly dose of EPO (18,000 IU). Logistic regression analysis also revealed that serum levels of EPO, gender or age were not associated with any significant variation of serum antibody level. High levels of serum antibodies to EPO are a risk factor for EPO resistance.

Serum albumin and mortality in patients with HIV and end-stage renal failure on peritoneal dialysis.

BACKGROUND: Peritoneal dialysis (PD) is an easily implementable dialysis modality in end-stage renal disease (ESRD). PD may improve access to renal replacement therapy in low- and middle-income countries; however, these countries have a higher prevalence of protein-energy wasting in patients and poorer socioeconomic conditions. We evaluated the effects of HIV infection on serum albumin levels in ESRD patients starting continuous ambulatory PD (CAPD) and mortality outcomes. METHODS: We conducted a single-center prospective cohort study of consecutive incident CAPD patients recruited from two hospitals in Durban, South Africa, from September 2012 to February 2015. Seventy HIV-negative and 70 HIV-positive ESRD patients were followed monthly for serum albumin levels and mortality events during the first 18 months of CAPD therapy. RESULTS: The HIV-positive cohort recorded 28 deaths (40%) among patients with a functional CAPD catheter at 18 months and 13 deaths (18.6%) in the HIV-negative cohort (p = 0.005). The mean serum albumin levels were lower in the HIV-positive cohort than in the HIV-negative cohort during the 18-month follow-up. The mean difference in serum albumin levels between the two cohorts was 4.24 g/L (95% confidence interval [CI] 2.02-6.46, p<0.001) at baseline and 3.99 g/L (95% CI 1.19-6.79, p = 0.006) at 18 months. HIV-positive status (adjusted regression coefficient -2.84, CI -5.00--0.67, p = 0.011), diabetes (adjusted coefficient -2.85; CI, -5.58--0.12; p = 0.041), and serum C-reactive protein and blood hemoglobin levels were independent predictors of serum albumin levels on multivariable linear regression. Baseline serum albumin <25 g/L (subdistribution-hazard ratio [SHR] 13.06, 95% CI 3.09-55.14, p<0.001) and CD4+ cell count <200 cells/µL (SHR 3.2, CI 1.38-7.45, p = 0.007) were independent predictors of mortality in our competing risk model. CONCLUSIONS: HIV infection can adversely affect serum albumin levels in ESRD patients managed with CAPD, while low baseline serum albumin levels and impaired immunity reliably predict mortality.

Continuous Ambulatory Peritoneal Dialysis in Patients with HIV and End-Stage Renal Failure.

♦ BACKGROUND: This study aimed to evaluate the differences in continuous ambulatory peritoneal dialysis (CAPD)-related outcomes according to human immunodeficiency virus (HIV) status of end-stage renal failure patients. ♦ METHODS: This prospective cohort study included 70 HIV-negative and 70 HIV-positive consecutive patients with renal failure who underwent dialysis with newly inserted Tenckhoff catheters between September 2012 and February 2015. Patients were followed up monthly at a central renal clinic for 1 year or until the primary endpoints of technique failure or death. ♦ RESULTS: Technique failure rates were similar (HIV-negative: 0.270 episodes/person-year; HIV-positive: 0.298 episodes/person-year; hazard ratio [HR], 1.09; 95% confidence interval [CI], 0.51 - 2.32; p = 0.822). However, there were fewer HIV-positive patients with complete 1-year follow-up with a patent catheter (42.9% vs 58.6% in the HIV-negative cohort; p = 0.063) owing to their higher all-cause mortality rate (0.55 vs 0.25 deaths/person-year, respectively; HR, 2.11; CI, 1.07 - 4.14; p = 0.031). Cluster of differentiation 4 count (CD4) < 200/µL (HR, 5.39; CI, 2.20 - 13.21; p < 0.001) and unsuppressed viral load (HR, 3.63; CI 1.72 - 7.67; p = 0.001) were associated with increased mortality hazards. Rates of first peritonitis were 0.616 (HIV-negative) and 1.668 (HIV-positive) episodes/person-year (HR, 2.38; CI, 1.46 - 3.89; p = 0.001). All-cause admission rates were 1.52 (HIV-negative) and 2.97 (HIV-positive) hospital admissions/person-year (HR, 1.66; CI, 1.12 - 2.48; p = 0.013). ♦ CONCLUSION: Although HIV-seropositive status of patients on CAPD did not adversely influence technique failure rates or patency at 1 year, uncontrolled HIV infection may be associated with increased relative risk of mortality and morbidity.

Association of Kidney Function and Waist Circumference with Uric Acid Levels in South Africans.

BACKGROUND: Recent evidence that hyperuricemia is associated with incident chronic kidney disease (CKD) provides a potential therapeutic target for CKD that has not been explored in Africans. With hyperuricemia and gout increasing globally, we sought to determine their prevalence in South Africans with varying kidney function levels. METHODS: This was a cross-sectional study of ambulatory adult patients presenting at a General Internal Medicine Outpatients Clinic between September 2012 and March 2014. Demographic, clinical, and laboratory data collected were analyzed using STATA11. Odds ratios (ORs) and 95% confidence intervals were determined using multivariable logistic regression with bootstrapping. RESULTS: There were 225/261 (86.2%) black/Africans, 31/261 (11.9%) Indian South Africans, 3/261 (1.1%) Caucasians, and 2/261 (<1%) mixed ancestry South Africans. Mean age was 51.3 ± 14.5 years. Median (interquartile range) estimated glomerular filtration rate (eGFR) was 71 (38) mL/min/1.73 m2 and 39.8% (104/261) of patients had CKD. Hyperuricemia prevalence was 43.7% (114/261) and increased from 16.7% in patients with eGFR ≥90 mL/min/1.73 m2 to 74.2% with eGFR <30 mL/min/1.73 m2 (P < 0.001). Gout prevalence was 5.4% (14/261), with equal distribution across eGFR categories (0.814). Factors independently associated with hyperuricemia were eGFR <90 [ORs 3.24 (1.15-9.14), 7.28 (2.26-23.49), and 7.88 (1.95-31.82) for eGFR 60-89.9, 30-60, and <30, respectively], albuminuria [2.32 (1.11-4.85)], and waist circumference [1.04 (1.01-1.06) per 1 cm increase]. In univariate and multivariable analysis, gout was positively associated with male gender and cardiovascular disease, while it was negatively associated with African ancestry, but none of these factors remained significant after bootstrapping; ORs 6.65 (0.64-69.24), 4.14 (0.61-28.07), and 0.18 (0.01-2.21), respectively. CONCLUSION: Hyperuricemia prevalence was high, with CKD and waist circumference being the strongest predictors. Gout was uncommon in black Africans. With population data lacking, screening high-risk individuals may provide insight into the burden of hyperuricemia and gout in South Africa.

Memory lymphocytes of young and old C57BL/6 mice express high levels of class I major histocompatibility complex (H-2 Kb) protein.

Memory lymphocytes play a central role in the secondary immune response. The concentration of memory lymphocytes increases with age. A high level of cell surface CD44 is a marker of memory lymphocytes compared to naïve lymphocytes which express a low level of CD44. Major histocompatibility complex (MHC) class I protein expression also increases with age. To explore a possible correlation between the expression of CD44 and MHC class I protein (Kb), peripheral blood lymphocytes from 27 C57BL/6 mice ranging in age from 3 months to 33 months were isolated by Ficoll-Hypaque gradient centrifugation. Single and double indirect immunofluorescence assays were then performed with rat IgG anti-CD44 and/or mouse IgG anti-Kb as first antibodies, and phycoerythrin (PE) labeled goat anti-rat IgG and/or fluorescein (FITC) labeled goat anti-mouse IgG as second antibodies. Cells were then analyzed by using a FACScan flow cytometer. As expected, the percentage of lymphocytes expressing high levels of CD44 (memory cells) increased significantly with age and the expression of Kb increased significantly with age. Interestingly, the expression of Kb in lymphocytes expressing high levels of CD44 (memory cells) was 72% more than in cells expressing low levels of CD44 (naive cells) regardless of age.

Effects of Sutherlandia frutescens extracts on normal T-lymphocytes in vitro.

Sutherlandia frutescens (SF), a popular traditional medicinal plant found in various parts of southern Africa, is used for treatment or management of HIV/AIDS and other diseases including cancer. However, its toxicity profile has not been fully established. The aims of this study were to examine the effects of 70% ethanol (SFE) and deionised water (SFW) extracts on normal isolated human T cells. An experimental study on normal human lymphocytes treated with doses SF extract doses ranging from 0.25 to 2.5 mg/ml. Untreated, vehicle-treated (Ethanol) and camptothecin (CPT) treated normal T cells were used as controls. Induction of cell death, changes in intracellular ATP, caspase-3/-7 activity and nuclear changes were analysed using flow cytometry, luminometry and nuclear staining (Hoechst) respectively. The highest concentration (2.5 mg/ml) of SFE extract induced significant necrosis (95%), depletion of ATP (76%), and inhibition of caspase-3/-7 activity (11%) following a 24 hour incubation period (p< 0.001). The 2.5 mg/ml concentration of SFW showed the same trend but were less effective (necrosis- 26%, ATP- 91%, & caspase-3/-7- 15%). These effects showed a time-dependence over 48 hours of incubation, with high doses of SFE extracts eliminating viable cells by necrosis, depleting ATP levels and decreasing caspase-3/-7 activity (p< 0.001). The activity of SFE extract was independent of ethanol. The SFW extract dilutions were less toxic than the SFE extracts. Significant DNA fragmentation as demonstrated by Hoechst staining was also seen over 48-hour incubation for high doses of both types of SF extracts. These results showed that although high concentrations of SF extracts can be toxic to normal T cells in vitro, SFW fractions were relatively safe for use.

The immunomodulatory effects of Sutherlandia frutescens extracts in human normal peripheral blood mononuclear cells.

Sutherlandia frutescens (SF) is one of the medicinal plants used as an immune booster in the treatment of chronic ailments such as HIV/AIDS and cancer. Limited data suggest that its efficacy is based on its regulatory effect on cytokines, the critical components of the immune response. In this study, we investigated the in vitro immunomodulatory effects of SF extracts on normal human peripheral blood mononuclear cells (PBMCs). An ELISA-based assay was used to assess the levels of expression of 12 cytokines in treated cells. An adenosine triphosphate (ATP) assay was used to assess cell viability in relation to cytokine secretion. SF ethanol extracts induced changes in cytokine secretion relative to the dose of the extract. Generally cytokine expression and secretion was low in concentration because were not stimulated with any endotoxin. The high SFE dose (2.5 mg/ml) significantly (p<0.001) decreased some cytokines including TNF-α and IL 1ß. Low doses of this extract (0.5 mg/ml) did not change TNF-α and IL 1ß secretion from the baseline (untreated cells). Changes in cytokine secretion of SFE treated cells tracked changes in ATP levels (cell viability). The SFW extract-induced changes in cytokine secretion were independent of cell viability. TNF-α was decreased (p<0.001) by the high dose of SFW extract while IL 1ß and IFNγ were increased (p<0.01) by the same dose. High doses decreased cell viability which was reflected in cytokine secretion. It is evident, from these results, that SF extracts can modulate cytokine secretion in unstimulated normal PBMCs in vitro. Further studies in animal models are recommended to advance understanding of this immunomodulatory activity.

Conversion of cyclosporine to sirolimus before 12 months is associated with marked improvement in renal function and low proteinuria in a South African renal transplant population.

OBJECTIVES: Avoidance of calcineurin inhibitor-associated nephrotoxicity has recently gained focus. To assess the impact of the conversion to sirolimus, we performed a retrospective audit on renal transplant patients switched to sirolimus at the Inkosi Albert Luthuli Central Hospital (South Africa) from 2003 until June 2007. MATERIALS AND METHODS: Medical records of transplant recipients were analyzed. Twenty-four-hour urine protein excretion and estimated glomerular filtration rates before initiation of sirolimus (baseline), and at their last clinic visit, were compared. Patients were then subcategorized according to their specific indications for switching to sirolimus. RESULTS: Thirty patients were included. Average follow-up was 25 months. Indications for use of sirolimus were group 1 (cyclosporine-induced biochemical toxicity, n=6); group 2 (chronic allograft nephropathy, n=6); group 3 (severe gum hypertrophy, n=9); group 4 (posttransplant diabetes, n=4); group 5 (calcineurin-inhibitor-induced histologic nephrotoxicity, n=2); and group 6 (calcineurin inhibitor-associated malignancy, n=3). Average urine protein excretion rate and estimated glomerular filtration rate before starting sirolimus were 0.44 -/+ 0.08 g/24 h and 50.1 -/+ 3.1 mL/min respectively, compared to 0.94 -/+ 0.2 g/24 h and 52.1 -/+ 4.8 mL/min, at an average follow-up of 25 months. On subgroup analysis, estimated glomerular filtration rate was increased/unchanged in groups 1 (47.3 vs 51.16 mL/min) and 4 (60.0 vs 60.0 mL/min) when compared to baseline, but decreased in groups 2 (47 vs 27.6 mL/min), 3 (51.3 vs 42.2 mL/min), 5 (54.0 vs 29.5 mL/min), and 6 (60.0 vs 56.5 mL/min). Combining the latter 2 groups, most patients (80%) received sirolimus within 1 year of transplant, whereas only 2 patients in the former groups (10%) received the drug within 1 year of transplant. CONCLUSIONS: Overall, sirolimus therapy was associated with improved estimated glomerular filtration rate, and also an increase in urine protein excretion rates. Maximum benefit was achieved when patients were switched to sirolimus within the first transplant year.

Transcription of major histocompatibility complex class I (Kb) and transporter associated with antigen processing 1 and 2 genes is up-regulated with age.

The transporter associated with antigen processing 1 and 2 (TAP1 and TAP2) genes belong to the ATP-binding cassette family of transporter genes. They provide peptides necessary for the assembly of major histocompatibility complex (MHC) class I molecules by transporting these peptides into the endoplasmic reticulum. As MHC class I protein expression increases with age, we have explored the effect of age on the transcription of MHC class I genes (Kb) and TAP1 and TAP2 genes in C57BL/6 mice. Blood and spleen lymphocytes were isolated from mice aged from 3 months to over 24 months. RNA was extracted and mRNA for Kb, TAP1, TAP2 was quantified using slot-blot hybridization followed by densitometry. There was a parallel age-related increase (1.5-fold) in blood lymphocyte mRNA of these genes from 3 months to 21 months. In mice over 24 months old there was a decrease in Kb and TAP1 mRNA, but an increase in TAP2 mRNA. In spleen lymphocytes an age-related increase in all three mRNA species occurred throughout life. While MHC class I and Tap genes underwent very similar age-related changes, MHC class I mRNA was about 50 times more abundant than either TAP1 or TAP2 mRNA.