A simple and cost-effective HPLC-UV method for the detection of levetiracetam in plasma/serum of patients with epilepsy.

A simple, fast and cost-effective method was developed and validated for the determination of levetiracetam (LEV) in plasma/serum of patients using high performance liquid chromatography (HPLC) with ultraviolet detection. The stability of LEV plasma/serum samples over time and in different blood collection tubes was evaluated. Serum/plasma samples were deproteinized by methanol spiked with the internal standard, gabapentin. HPLC was carried out on a Venusil XBP C18 , 250 × 4.6 mm, 5 µm column, at a flow rate of 1.0 mL/min and with mobile phase consisting of 50 mm potassium dihydrogen phosphate-acetonitrile at a pH of 5.5. The UV detector was set at 205 nm and 10 µL was injected. Total runtime was 15 min. Calibration curves were linear (correlation coefficient = 0.999) over a concentration range of 1-60 µg/mL. Relative standard deviation values for both the inter-day and intra-day precision and accuracy were <5% for the concentration range. The influence of different collection tubes and the effect of time on the stability of LEV was investigated. These factors may cause inaccuracies owing to drug-protein binding and interference in the matrix. This method is simple, fast, cost-effective, reliable and accurate with minimal sample preparation for daily routine use in therapeutic drug monitoring.

Self-reported work-related musculoskeletal injuries and isometric handgrip strength.

BACKGROUND: Musculoskeletal injuries are common in car assembly workers, causing significant morbidity. There is evidence that increased muscle strength reduces musculoskeletal injury risk and that handgrip strength is a predictor of general muscle strength. Aims To investigate whether isometric handgrip strength (IHGS) is a predictor of increased musculoskeletal injury risk in automotive assembly workers. METHODS: We measured IHGS in a cross-sectional survey of final assembly line workers in an automotive manufacturing plant. Weak handgrip was defined as two standard deviations below the normative mean value for age and gender. Self-reported musculoskeletal injury/accident data and recent musculoskeletal symptoms were recorded using a modified Nordic musculoskeletal questionnaire. RESULTS: Of 445 final assembly line workers, 206 (46%) participated. Sixty participants (29%) reported a musculoskeletal injury/accident at work in the preceding 5 years. Mean IHGS in this population was lower than international normative values. Odds ratios for self-reported injury (adjusted for gender and recent musculoskeletal symptoms) were 0.62 (95% CI 0.28-1.35) for left weak handgrip and 0.57 (95% CI 0.31-1.1) for right weak handgrip. CONCLUSIONS: In this study, weak handgrip did not predict increased risk of musculoskeletal injuries/accidents.

Beliefs of grade six learners' regarding adolescent pregnancy and sex.

Escalating adolescent pregnancy and risky sexual behaviour is becoming more common amongst young adolescents and especially amongst black adolescents in South Africa. Statistics confirm that South African adolescents as young as fourteen are already sexually active and become pregnant. The decision to become sexually active with resulting adolescent pregnancy whether planned or not, are directly influenced by the teenager's beliefs. A person's beliefs consist of a person' own individual beliefs or attitude as well as what the individual subjective norm which the individual perceive as other people's beliefs regarding the same object of reason. The aim of the study was to describe the attitude of black grade six learners under the age of fourteen, towards adolescent pregnancy and sex. A quantitative descriptive research design was used. Results were clustered according to demographic variables as well as beliefs that consist of attitude and subjective norm. Findings provided insight into the beliefs of grade six learners regarding sex and adulthood, the roll of peer pressure, relationships of adolescent parents, social interaction of teenage parents, ability of adolescent parent's ability to provide in the needs of the baby, the adequacy of a child support grant to raise a baby as well as the levels of education of adolescent parents. This article provide a detailed reflection on these results and propose off a doll parenting intervention strategy as means of modification of attitude and subjective norms of grade six learners in order to alter sexual behaviour.

The mammalian metabolite, 2-methoxyestradiol, affects P53 levels and apoptosis induction in transformed cells but not in normal cells.

The endogenous metabolite, 2-methoxyestradiol (2ME), is an inhibitor of tubulin polymerization and is therefore toxic to dividing fast-growing tumor cells. Transformed cells are not equally susceptible to the effects of 2ME. In this study the effects of 1-2 microM doses of 2ME on cell cycle progression, apoptosis induction and on p53 levels were evaluated using flow cytometry in cells with different p53 status. No effect of 2ME was seen in normal human skin fibroblast strain HSF43 with wild-type (wt) p53. However, in SV40 T antigen transformed HSF43 cells (line E8T4), 2ME caused a prominent G2/M arrest, with subsequent micronuclei formation followed by apoptosis. Increased p53 levels were present in the G2/M cells. Our results suggest that 2ME, being a microtubule poison, may release the bound p53 from T antigen, and that this p53 may enhance the apoptotic effects. Two lymphoblast cell lines derived from the same donor, TK6, expressing low levels of wt p53, and WTK1, expressing high levels of mutant p53, showed similar moderate responses to 2ME at 37 degrees C. The effects included enhanced apoptosis and a modest G2/M block. No increase in p53 levels was seen. However, at the permissive temperature of 30 degrees C marked increases in apoptosis and a prominent G2/M-phase block, similar to that seen in the E8T4 cells, were present in the WTK1 cells, indicating that the high levels of mutant p53 have now become functional, enhancing the apoptotic effects initiated by 2ME.

Effects of gamma-linolenic acid and arachidonic acid on cell cycle progression and apoptosis induction in normal and transformed cells.

The effects of arachidonic acid (AA) and gamma-linolenic acid (GLA) on cell cycle progression and apoptosis induction, using flow cytometry, were compared on normal human skin fibroblasts, strain HSF43 with wild type (wt) p53, large T antigen transformed HSF43 cells (line E8T4) with non functional p53, and on two lymphoblast cell lines, TK6 with wt p53 and WTK1 with mutant p53. AA and GLA caused similar dose (50, 75 and 100 microg/ml AA and GLA) and time dependent (24 and 48 h) induction of apoptosis in each cell line. The degrees of the response of the four cell lines were, however, different. The normal HSF43 cells were most resistant against apoptosis induction and the WTK1 cells most susceptible. The apoptosis induction appeared to be independent of functional p53. Cell cycle progression was also similarly affected by AA and GLA in the two cell types. In the fibroblast type cells (HSF43 and E8T4) S- and G2/M-phase arrests were evident after 48 h exposure to AA and GLA, and in the lymphoblast cell lines (TK6 and WTK1) the cells were arrested in the G1-phase.

17beta-Estradiol metabolites affect some regulators of the MCF-7 cell cycle.

The activity of p34(cdc2) plays a key role in the regulation of the eukaryotic cell cycle. Another cell cycle associated molecule is PCNA. We investigated the effects of 2-hydroxy-17beta-estradiol, a cell proliferator, and 2-methoxy-17beta-estradiol, a potent inhibitor of cell growth, on the levels and activity of p34(cdc2) and on the levels of PCNA, as well as on protein phosphorylation in MCF-7 cells. 2-Hydroxyestradiol increased p34(cdc2) activity at G1/S and elevated PCNA levels during S-phase. 2-Methoxyestradiol caused unscheduled activation of p34(cdc2) in S-phase and decreased levels of p34(cdc2) and PCNA during G2/M. We conclude that 2-hydroxy- and 2-methoxyestradiol have definite, though different regulatory functions during the cell cycle.

The induction of apoptosis in human cervical carcinoma (HeLa) cells by gamma-linolenic acid.

A high concentration (50 micrograms/ml) of gamma-linolenic acid (GLA) induced morphological lesions typical of apoptosis, as well as DNA fragmentation, in HeLa cells. A lower concentration of GLA (20 micrograms/ml), caused an increased proliferating cell nuclear antigen (PCNA) labelling, with 92.7% cells positive, compared to 27.7% at a concentration of 50 micrograms/ml GLA. In correlation with these results, the number of cells with degraded DNA below the G0/G1 peak increased significantly in the 50 micrograms/ml GLA-treated cells, but increased only slightly in cells exposed to the lower level of GLA. The high levels of PCNA induced by 20 micrograms/ml GLA, in both G1 and S phases, may indicate a state of DNA repair synthesis, whilst at the higher concentration of GLA, most of the cells became apoptotic. Since apoptosis is associated with the deregulation of c-Myc expression, and as the Raf-1-MAP kinase cascade activates the expression of c-Myc and c-Jun, we investigated the effects of 20 and 50 micrograms/ml GLA on the Raf-1, c-Myc and c-Jun levels, and on the activity of MAP kinase. The results showed that 50 micrograms/ml GLA lowered the activity of MAP kinase. As expected with the decreased MAP kinase activity in the cells exposed to the higher level GLA, the c-Jun levels were also lowered. The levels of c-Myc, however, were increased. It is therefore possible that the deregulated expression of c-Myc in the HeLa cells exposed to the high level of GLA (50 micrograms/ml) may contribute to the induction of apoptosis in HeLa cells.

Maternal deaths at Pelonomi Hospital, Bloemfontein, 1980-1985. A survey of 81 consecutive cases.

From 1 January 1980 to 31 December 1985, 81 maternal deaths occurred at Pelonomi Hospital, Bloemfontein; these were classified as direct obstetric-related (74 cases), indirect obstetric-related (6) and non-obstetric (1). The overall maternal mortality rate was 2.87 per 1000 deliveries, including deaths related to abortion and ectopic pregnancy, but excluding the non-obstetric death. Among booked patients the maternal mortality rate was 0.32 and among unbooked patients 11.13 per 1000 deliveries. The maternal mortality rate for patients from the Bloemfontein area was 0.72 per 1000. Puerperal sepsis and postabortal sepsis accounted for 45.7% of the deaths. Avoidable factors were considered to be present in 65 cases (80.2%).