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Biomedical subjects

D Moodley

Publications and source records attributed to D Moodley.

At least 19 recordsLinked to original sources

Cytotoxicity of a dentine bonding agent on four different cell-lines.

UNLABELLED: The aim of this study was to compare the cytotoxicity of a recently available dentine bonding agent on four different cell-lines (three human dental pulp fibroblast cell-lines and one mouse 3T3 fibroblast cell-line). METHODOLOGY: Three human dental pulp cell-lines from 3 different donors and one established 3T3 mouse cell-line were grown and sub-cultured. Cell viability following exposure to Scothbond was then compared to a similar number of controls using the MTT assay. RESULTS: Scotchbond 1 was cytotoxic to all four cell-lines. 3T3 cells showed a survival rate of about 60% as compared to two of the human dental pulp cells which showed a significantly lower survival rate (p<0.05, Kruskal-Wallis Multiple-Comparison Test). CONCLUSION: These findings indicated that is cytotoxic to both human pulp and 3T3 cell-lines. In general, the human pulp cell-lines showed higher sensitivity than the 3T3 cell-lines. CLINICAL SIGNIFICANCE: Scotchbond 1 cannot be recommended for direct pulp capping techniques and care should be taken when using this dentine bonding agent in cavities where the remaining dentine layer is minimal.

3T3 Cells↗

Management of human immunodeficiency virus infection in pregnancy.

The HIV global epidemic is having a devastating effect on women of reproductive age; women aged 15-24 years are 2.5 times more likely to be infected than young men in the same age group. Further, mother-to-child transmission (MTCT) accounts for almost two-thirds of the new infections that occur in children world-wide, annually. MTCT of HIV-1 varies widely and is dependent on obstetric practices, mode of delivery, breastfeeding, and the level of the viral load in the mother. Antiretroviral therapy (ARV) in pregnancy is prescribed for two main reasons: (i) women who need ARV medication for their own health; (ii) women who do not need treatment, or do not have access to treatment are offered prophylaxis to prevent MTCT, using one of a number of ARV regimens known to be effective. HIV infection is also associated with significant maternal morbidity and mortality. Clinicians caring for HIV-infected women need to update their knowledge continuously to provide optimal care.

Adolescent↗

Vertical transmission of Mycobacterium tuberculosis in KwaZulu Natal: impact of HIV-1 co-infection.

BACKGROUND: Increases in perinatal TB have paralleled the exacerbation of the TB epidemic in KwaZulu Natal. The exact risks for vertical transfer of Mycobacterium tuberculosis (VTRTB) to the baby are unknown, as is the impact of HIV-1 co-infection, which frequently accompanies maternal TB disease in the region. DESIGN: Prospective case series study of 82 HIV-1-infected and 25 non-infected pregnant mothers, King Edward VIII Hospital, KwaZulu Natal, South Africa. RESULTS: Perinatal mortality in HIV-1/TB diseased mothers was 85/1000 and associated with maternal anaemia (P = 0.02); 46% of newborns were premature, 66% low birth weight and 49% intrauterine growth restricted. These were significantly higher than overall hospital rates (P < 0.01, OR 4.8, 95%CI 3.2-7.0). Sites of detection of maternal TB, distribution of bacteriologically-proven TB, obstetric comorbidity and perinatal morbidity were similar in HIV-1-infected and non-infected mothers. VTRTB was detected in 16 newborns (16%), occurring similarly in bacteriologically-proven and suspected maternal TB disease, with no difference between HIV-1-infected and non-infected mothers. Eleven newborns with VTRTB were HIV-1 exposed; 64% acquired HIV-1 and died from rapidly progressive disease by 10 months of age. HIV-1-infected mothers and their exposed newborns had significantly lower CD4 counts. No association between perinatal maternal viral load, CD4 count or VTRTB was detected. CONCLUSION: Mothers with TB disease in pregnancy are at risk for significant perinatal morbidity, mortality and VTRTB.

AIDS-Related Opportunistic Infections↗

Cytotoxicity of two concentrations of a dentine bonding agent on mouse 3T3 and human pulp fibroblast cell-lines.

The objective of this study was to investigate the cytotoxicity and effect of concentration of a recent dentine bonding agent on a mouse fibroblast cell-line (3T3) and four different human pulp fibroblast cell-lines. A mouse fibroblast 3T3 cell-line and 4 different human pulp fibroblast cell lines were used. The cells were grown and subcultured using standard conditions. For the testing of the cytotoxicity of the bonding agent (Prime & Bond NT) it was extracted with DMEM medium and the different cell-lines were exposed to the extraction. The MTT assay was used to establish the number of viable cells. For all 5 pairs (control vs test sample) of cell-lines for 4 microl bonding agent, the mouse and only two of the four human cell-lines showed a statistical significant difference (p < 0.05; Mann-Whitney) due to the cytotoxicity of the bonding agent. However, all 5 pairs showed significant differences (p < 0.05) at the 8 microl concentration level. To investigate the relative effect of concentration (4 microl compared to 8 microl) of a chemical agent (we used a bonding agent, Prime & Bond NT) on the cytotoxicity of these 5 cell-lines, the Kruskal-Wallis Multiple-Comparison test was used on the standardised medians. Statistically significant differences (p < 0.05) were found for various cell-lines between both concentration levels. In general, the tested bonding agent, at a higher concentration level, has an increased cytotoxic effect on all 5 cell-lines. About 69% (median) of 3T3 cells stayed viable with 4 microl (0.94cm2/ml) bonding agent and 61% when 8 microl was used. For the pulp cells the percentages were between 91% and 72% for 4 microl and between 77% and 24% for 8 microl. These findings indicated that the bonding agent Prime and Bond NT was cytotoxic and that different human pulp fibroblast cell-lines should be used in the cytotoxic testing of dental materials and secondly, if only one concentration is to be used it should be in the higher range of that suggested by the ANSI/ADA document (2000, Revision) for recommended standard practices for biological evaluation of dental materials.

3T3 Cells↗

Dentine bonding agents--a review of adhesion to dentine.

Although constant innovation has improved the performance of dentine adhesives, the true mechanism of resin adhesion to dentine is still not clear and the optimal dentinal surface pretreatment has yet to be determined for adhesive integrity of resin to dentine. The integrity of the collagen fibrils left exposed upon acid etching seem to play a major role in the mechanism of adhesion and intermingling of the adhesive monomers with the filigree of collagen fibers or hybrid layer should be considered the paramount dentine bonding mechanism. Definite trends are emerging but more research is necessary before general conclusions can be made about the functions of different bonding agents. The all-in-one self-etching/self-primer bottle systems are relatively new to the market and need more research before they can be advocated as the agent of choice.

Acid Etching, Dental↗

T-helper cell responses to HIV envelope peptides in cord blood: protection against intrapartum and breast-feeding transmission.

BACKGROUND: Acquired HIV-specific cell-mediated immune responses have been observed in exposed-uninfected individuals, and it has been inferred, but not demonstrated, that these responses constitute a part of natural protective immunity to HIV. This inference was tested prospectively in the natural exposure setting of maternal-infant HIV transmission in a predominantly breast-fed population. METHODS: Cord blood from infants of HIV-seropositive women in Durban, South Africa, were tested for in vitro reactivity to a cocktail of HIV envelope peptides (Env) using a bioassay measuring interleukin-2 production in a murine cell line. Infants were followed with repeat HIV RNA tests up to 18 months of age to establish which ones acquired HIV-infection. RESULTS: T-helper cell responses to Env were detected in 33 out of 86 (38%) cord blood samples from infants of HIV-seropositive women and in none of nine samples from seronegative women (P = 0.02). Among infants of HIV-seropositive mothers, three out of 33 with T-helper responses to Env were already infected before delivery (HIV RNA positive on the day of birth), two were lost to follow-up, and none of the others (out of 28) were found to be HIV infected on subsequent tests. In comparison, six out of 53 infants unresponsive to Env were infected before delivery, and eight out of 47 (17%) of the others were found to have acquired HIV infection intrapartum or post-partum through breast-feeding (P = 0.02). CONCLUSIONS: T-helper cell responses to HIV envelope peptides were detected in more than one-third of newborns of HIV-infected women; no new infections were acquired by these infants at the time of delivery or post-natally through breast-feeding if these T-helper cell responses were detected in cord blood.

Animals↗

Growth in early childhood in a cohort of children born to HIV-1-infected women from Durban, South Africa.

This study describes growth in a cohort of black South African children born to HIV-1-infected women in Durban. Children born to HIV-1-seropositive women were followed up from birth to early childhood. At birth and at each visit, growth parameters were measured. Mean Z-scores were calculated for weight-for-length, weight-for-age and length-for-age and, if they were low, the children were regarded as wasted, malnourished or stunted, respectively. At the end of the study, there were 48 infected and 93 uninfected children. There were no significant differences between the two groups at birth. Thereafter, the infected group was found to have early and sustained low mean Z-scores for length-for-age and weight-for-age but not for weight-for-length. The means reached significance at ages 3, 6 and 12 months for length and at 3, 6 and 9 months for weight. Infected children who died early had more severe stunting, wasting and malnutrition than infected children who survived. Infected children born to HIV-positive women have early and sustained stunting and are malnourished but not wasted. Children with rapidly progressive disease have both stunting and wasting and are more severely affected. Early nutritional intervention might help prevent early progression or death in HIV-infected children, particularly in developing countries without access to anti-retroviral therapy in state hospitals.

Anthropometry↗

Severe, rapidly progressive human immunodeficiency virus type 1 disease in newborns with coinfections.

AIM: To describe a severe form of rapidly progressive HIV-1 infection manifesting in the neonatal period. METHOD: Prospective cohort study, King Edward VIII Hospital, Durban, South Africa. HIV-1-exposed neonates with hepatosplenomegaly, lymphadenopathy or persistent pneumonia within the first 28 days of life were investigated for perinatal infections. Confirmation of neonatal HIV-1 infection, HIV-1 subtype and clinical outcomes were studied. RESULTS: Twenty-three (72%) of 32 symptomatic HIV-1-exposed neonates recruited at a mean of 15.2 days were HIV-1-infected. HIV-1 infection was detected in 5 patients who were tested within 48 h of birth, confirming congenital infection. Congenital infection was not excluded in any case. Median neonatal viral load at recruitment was 471,932 copies/ml and median CD4 was 777 cells/mm3. The predominant clinical presentation was growth retardation and prematurity. Perinatal infections detected included: tuberculosis (8), syphilis (6) and cytomegalovirus (10). All of the neonates with perinatal tuberculosis were HIV-1-coinfected. Maternal and neonatal viral load and CD4 at recruitment were not statistically different between the groups with tuberculosis vs. other coinfections. Gag gene sequence analysis confirmed closely aligned HIV-1 subtype C in mothers and neonates. Nineteen (83%) died by 9 months, with a mean age at death of 3.5 months. CONCLUSIONS: A distinct group of HIV-1-infected babies may clinically manifest in the neonatal period with perinatal coinfections, subsequent rapidly progressive HIV-1 and early death.

AIDS-Related Opportunistic Infections↗

An evaluation of on-site testing for syphilis.

One of the major obstacles to the eradication of perinatal transmission of syphilis is the delay in obtaining the results of syphilis serological tests. The availability of on-site syphilis testing lead to this study which attempted to evaluate on-site syphilis testing performed by nursing staff. The seroprevalence of syphilis by laboratory rapid plasma reagin (RPR) was 8.2% (n=42). Twenty-one of the 42 women were correctly identified by the on-site test. The overall sensitivity of on-site testing was 50% [95% confidence interval (CI)=34.4-65.6]; specificity of 90.9% (95% CI=87.8-93.2). The on-site test correctly identified as uninfected 429 of the 471 women reported as RPR negative by the laboratory, giving a specificity of 91.1% (95% CI=88.1-93.4). The results of the study show that on-site RPR test had a sensitivity of 75% in respect of the clinically important titres of > or = 1:8. The specificity of the on-site test was 91.1% and on-site testing only failed to detect syphilis in those patients with titres of 1:1 and 1:2. On-site testing is a practical and cost-effective option to prevent congenital syphilis, in settings of a high prevalence of syphilis and using skilled testers.

Adolescent↗

Pharmacokinetics of zidovudine and lamivudine in neonates following coadministration of oral doses every 12 hours.

A phase I, repeat-dose, open-label study was conducted to determine the pharmacokinetics and safety of zidovudine and lamivudine, coadministered orally every 12 hours, in 16 neonates whose mothers were infected with human immunodeficiency virus type 1 (HIV-1). The prospective mothers had been stabilized on a zidovudine/lamivudine regimen since week 36 of pregnancy to prevent mother-to-child transmission of HIV. During 1 week postpartum, the mothers received zidovudine 300 mg plus lamivudine 150 mg every 12 hours and breastfed. Neonatal treatment was initiated 12 hours following birth with 4 mg/kg of zidovudine suspension plus 2 mg/kg of lamivudine solution every 12 hours; this regimen was continued for 1 week. Between days 1 and 7 of neonatal treatment, the neonatal oral clearance (CL/F) of zidovudine and lamivudine increased by 2-fold (p < 0.001) and 1.6-fold (p = 0.004), respectively, possibly reflecting maturation of intestinal hepatic and renal function occurring during the first week of life. Day 7/day 1 ratios for exposure (area under the serum concentration-time curve [AUC]) and maximum observed serum concentration (Cmax) were 0.48 and 0.63, respectively, for zidovudine and 0.64 and 0.73, respectively, for lamivudine. At the time of delivery, the geometric mean cord/maternal concentration ratio was 1.24 for zidovudine and 1.12 for lamivudine, indicating free passage of each drug across the placenta. The maternal and neonatal treatment regimens were well tolerated. The results of this study confirm that in the neonate, a convenient regimen combining zidovudine 4 mg/kg and lamivudine 2 mg/kg, administered orally every 12 hours, provides zidovudine serum exposure very similar to that reported with the standard neonatal zidovudine regimen of 2 mg/kg every 6 hours, as well as lamivudine serum exposure within the range reported in adults receiving lamivudine 150 mg twice a day and children receiving 4 mg/kg twice a day.

Adult↗

Interferon-gamma and interleukin-10 production among HIV-1-infected and uninfected infants of HIV-1-infected mothers.

Immunologic consequences of exposure to HIV-1 in utero are still poorly understood. This study investigates relationships between type-1 [interferon-gamma (IFN-gamma)] and type-2 (IL-10) cytokine production and maternal-infant HIV-1 transmission. Cord blood leukocytes from deliveries of 71 HIV-1-infected and 11 uninfected mothers were tested for in vitro IFN-gamma and IL-10 production after phytohemagglutinin (PHA) stimulation. The infants of these HIV-1-infected mothers were followed prospectively after birth to determine HIV vertical transmission, and IFN-gamma and IL-10 production was measured again at 6 mo. Median PHA-stimulated IFN-gamma production was 210 pg/mL in cord blood cells from infected and 73 pg/mL from uninfected mothers (p = 0.12), and median PHA-stimulated IL-10 production was 491 pg/mL in cord blood cells from infected and 161 pg/mL from uninfected mothers (p = 0.004). PHA-stimulated IFN-gamma and IL-10 production alone were not significantly associated with transmission, but relationships between the two cytokines differed among infected and uninfected infants of HIV-1-infected mothers. PHA-stimulated IFN-gamma and IL-10 production was positively correlated among infected (r = 0.7, p = 0.12 in cord blood and r = 0.66, p = 0.03 at 6 mo) but not uninfected infants, and stronger relative production of IFN-gamma to IL-10 was observed among exposed uninfected than among infected infants (p = 0.04). Exposure in utero to HIV-1 may augment production of IL-10 detectable in fetal cord blood. Stronger relative production of IFN-gamma to IL-10 in cord blood cells from infants of HIV-1-infected mothers may be associated with protection against perinatal HIV infection.

Female↗

In vitro evaluation of two adhesive systems used with compomer filling materials.

AIM: To evaluate and compare the in-vitro shear bond strength, microleakage and dentine-restorative interface of a self-etching/self-priming dentine bonding agent with a three-component dentine-bonding agent. METHOD: For shear bond strength (SBS) testing 30 non-carious human molars were used of which 15 were tested with Non-Rinse Conditioner (NRC)/Prime&Bond NT (PBNT) and Dyract AP and 15 were tested with Scotchbond Multi-Purpose Plus (SBMP) and F2000. For the microleakage evaluation cavity preparations were made on the facial surfaces of 30 non-carious human premolars of which 15 were restored with NRC and PBNT with Dyract AP and 15 were restored with SBMP and F2000. The dentine-restorative interface was examined through a confocal scanning laser microscope (CSLM). RESULTS: The mean SBS of PBNT and SBMP were 12.8 and 18.1 MPa, respectively. The microleakage scores showed NRC/PBNT leaked on the dentine side in 13 of the 15 specimens examined. On the enamel side two of the 15 specimens showed microleakage. With SBMP no microleakage was observed on either enamel or dentine sides. The CSLM images show clear resin tag and hybrid layer formation for both the materials examined although SBMP showed better and deeper penetration into the dentine with longer resin tags. SBMP showed resin tags measuring about 150 mm while the hybrid layer measured about 5 mm. The length of the resin tags as well as the thickness of the hybrid layer for PBNT were 20 mm Pounds and 2 mm Pounds, respectively. CONCLUSIONS: The acid-etch technique of SBMP produced higher bond strength and no microleakage when compared with the self-etching/self-priming 'non-rinse technique' of NRC with PBNT. Thus it can only be speculated that SBMP should be the superior in the clinical situation.

Compomers↗

Tissue kallikrein and kinins in renal disease.

The renal kallikrein-kinin system is involved in sodium and water homeostasis, blood pressure regulation and inflammation. Tissue kallikrein and kinin levels were measured in the urine of patients with renal disease and in the urine of living related kidney donors prior to uninephrectomy who served as controls. Tissue kallikrein and kinin B1 and B2 receptors were immunolocalised by confocal microscopy in renal biopsy material from patients with renal disease and controls (fresh autopsy material and normal kidney tissue from nephrectomies for malignancy). Urinary tissue kallikrein excretion was significantly decreased in patients with mild renal disease (16.6 +/- 6.7 ng tissue kallikrein (TK)/ng protein; p < 0.05) and more markedly so (1.8 +/- 0.7 ng TK/microg protein; p < 0.01) in patients with severe renal failure requiring dialysis compared to normal controls (78.9 +/- 31.7 ng TK/microg protein). Basal kinin values were unchanged in patients with renal disease (14 +/- 0.8 ng/ml) compared to controls (13.3 +/- 0.56 ng/ml). In control kidney tissue kallikrein was immunolocalised in the distal connecting tubules and collecting ducts whereas decreased immunolabelling was observed with renal disease. Kinin B2 receptor labelling was present in the entire nephron in the normal control kidney but was reduced with renal disease. While kinin B1 receptor immunolabelling was not observed in the control kidneys, labelling of distal tubules and collecting ducts was noted in renal disease, suggesting an upregulation of B1 receptors in renal parenchymal disease.

Adolescent↗

HIV type 1 V3 domain serotyping and genotyping in Gauteng, Mpumalanga, KwaZulu-Natal, and Western Cape Provinces of South Africa.

More than 20.8 million people are living with HIV/AIDS in sub-Saharan Africa, with southern Africa the worst affected area and accounting for one of the fastest growing AIDS epidemics worldwide. Samples from 81 patients, including 25 from KwaZulu-Natal, 26 from Gauteng, 5 from Mpumalanga, and 25 from Western Cape Province, were serotyped using a competitive V3 peptide enzyme immunoassay (cPEIA). Viral RNA was also isolated from serum and the V3 region amplified by reverse transcriptase polymerase chain reaction (RT-PCR) to obtain a 240-bp product for direct sequencing of 29 samples. CLUSTAL W was used to make multiple sequence alignments. Distance calculation, tree construction methods, and bootstrap analysis were done using TREECON. Subtype C-like V3 loop sequences predominate in all provinces tested in South Africa. Discordant sero- and genotype results were observed in one patient only. The correlation between sero- and genotyping was 96% (24 of 25) in KwaZulu-Natal and 100% in Gauteng and Mpumalanga. In Western Cape Province 18% of patients were identified as sero/genotype B and 82% as sero/genotype C. Our data show that results of the second-generation V3 cPEIA correlated well with V3 sequencing and would be a rapid and affordable screening test to monitor the explosive southern African HIV-1 epidemic.

Adolescent↗