A phase I trial of hypoxoside as an oral prodrug for cancer therapy - absence oftoxicity

No Abstract

The role of plant sterols in dietary control of high plasma cholesterol levels

[No abstract available]Editoria

Analysis of human sperm membrane antigens reacting with sera from antisperm antibody positive and negative patients by Western blotting

Immunological infertility is thought to be caused by the binding of antibodies to 'fertility-related' antigen(s) on the sperm membrane. We compared antibody profiles in sera from 20 ASA(+) and ASA(-) men, using a sperm membrane extract as an antigen. Antigens were separated by SDS-PAGE under reducing conditions. The patients were classed as ASA(+) by the MAR (>50%), d-IBT (>20%) and TAT (>1:64). The results showed that immunoreactive bands in both the ASA(+) and ASA(-) groups were heterogenous and included bands covering the whole molecular weight range. Statistical analysis showed significantly more patients in the ASA(+) group having immunoreactive bands at molecular weights of 32 Kd (P = 0.006) and 79 Kd (P = 0.02) when compared to the ASA(-) group. In the ASA(-) group significantly more patients had reactive bands at 81 Kd (P = 0.01) when compared to the ASA(+) group. The 32 Kd antigen reacted only with sera from ASA(+) patients. We conclude that differences exist between the ASA(+) and ASA(-) groups when this extraction method is used and that the isolation and purification of the 32 Kd protein may justify further investigation.Articl

TUNEL assay and SCSA determine different aspects of sperm DNA damage

Summary For the determination of sperm DNA damage, different assays are used. However, no further distinction is made and the literature generally speaks about DNA damage. Thus, this study aimed at comparing the sperm chromatin structure assay (SCSA) and the TUNEL assay. In 79 patients, sperm DNA damage was determined flow cytometrically using the SCSA and the TUNEL assay. Moreover, normal sperm morphology was evaluated according to strict criteria. A statistical comparison of the two methods was performed using standard correlations, Bland and Altman plots, Passing-Bablok regressions and concordance correlation. Results show a significant difference between P- and G-pattern morphology only for the mean channel fluorescence of the SCSA. Spearman's rank correlations between the different parameters of both assays, SCSA and TUNEL, revealed significant associations between the parameters of the assays. However, when applying Bland and Altman plots, Passing-Bablok regression and concordance correlation results showed that these methods are not comparable. These different techniques determine different aspects of sperm DNA damage, i.e. 'real' DNA damage for the TUNEL assay and 'potential' DNA damage in terms of susceptibility to DNA denaturation for the SCSA. Thus, one should clearly distinguish between the different assays, not only practically and methodologically but also linguistically. © 2010 Blackwell Verlag GmbH.Articl

Skin Permeability of Different Terbinafine Hydrochloride Formulations

The objectives of the study are to determine the in vitro permeability of different terbinafine hydrochloride formulations through human skin and to measure the respective concentrations of each formulation within the exposed skin tissue. The permeation of three commercially available 1% terbinafine hydrochloride formulations and two terbinafine hydrochloride solutions of 10 and 20 mg/ml through human skin was investigated using an in vitro continuous flow-through perfusion system. The terbinafine hydrochloride retained in the skin was extracted and analysed. The terbinafine hydrochloride from the different formulations readily diffused into the skin tissue. However, no flux values for any of the terbinafine hydrochloride formulations through the skin into the receptor fluid were found. The mean terbinafine hydrochloride concentrations in the skin after 24 h exposure to the three commercial formulations were 3.589, 1.590 and 4.219 μg/ml respectively. The mean terbinafine hydrochloride concentrations in the skin after 24 h of exposure to the terbinafine hydrochloride solutions (PBS/Methanol 1:1) of 10 and 20 mg/ml were 85.280 and 154.680 μg/ml respectively. The mean terbinafine hydrochloride concentration in the skin exposed to the 10 mg/ml PBS/Methanol solution was higher than those from the three commercial formulations. Terbinafine seems to accumulate in skin/bind to the skin, rather than to diffuse through the skin into the receptor compartment. This unique pharmacokinetic property of terbinafine hydrochloride may enhance its efficacy as topical antifungal and reduce systemic side effects

A novel approach for the selection of human sperm using annexin V-binding and flow cytometry

Objective: To develop a method whereby sperm with phosphatidylserine externalization can be separated from those without this feature. Because annexin V binds phosphatidylserine, this study is using this feature to select functional spermatozoa. In addition, the relationship between annexin V binding in human spermatozoa and normal sperm morphology according to strict criteria was to be assessed. Design: Prospective study. Setting: Department of Obstetrics and Gynaecology of Stellenbosch University at Tygerberg Academic Hospital, Tygerberg, South Africa. Patient(s): Semen from 14 healthy sperm donors. Exclusion criterion was the presence of less than 20 × 106/mL total motile spermatozoa in the original sample. Main Outcome Measure(s): Annexin V-negative sperm, annexin V-positive sperm, normal sperm morphology. Intervention(s): An aliquot of a semen sample after double density gradient centrifugation was incubated with annexin V fluorescein isothiocyanate conjugate (FITC). Cell fluorescence signals were determined using a FACScalibur flow cytometer equipped with a FACSSort fluidic sorting module. The sorting procedure delivered two sperm subpopulations: annexin V-negative and annexin V-positive. Morphology slides were made and the sperm morphology was assessed according to strict criteria. Result(s): There was a significant enrichment of annexin V-negative sperm as well as morphologically normal sperm in the annexin V-negative subgroup after separation with flow cytometry. The percentage of morphologically normal sperm increased from 8.3% in the control to 11.9% in the annexin V-negative fraction, whereas the percentage of annexin V-positive sperm decreased to 5.7%. Conclusion(s): The annexin V-negative sperm subpopulation had morphologically superior quality sperm compared to annexin V-positive sperm. It is important to select morphologically normal sperm during intracytoplasmic sperm injection (ICSI) as it may contribute to increased implantation and pregnancy rates (PR). © 2009 American Society for Reproductive Medicine.Articl

Use of specific monoclonal antibodies to secretory IgA for the detection of spermatozoal antibodies in serum and seminal plasma by enzyme-linked immunosorbent assay

The role of anti-sperm secretory IgA has recently received attention since some workers feel it plays an important role in the prognosis of the immunologically infertile couple. Current methods used in our laboratory cannot separately detect anti-sperm secretory IgA. An enzyme-linked immunosorbent assay (ELISA) utilizing specific monoclonal antibodies to secretory IgA was used to detect anti-sperm secretory IgA as well as anti-sperm monomeric IgA and IgG in serum and seminal plasma of a sperm-antibody-positive (ASA+) and sperm-antibody-negative (ASA-) group of men. Results showed significantly raised serum levels in the ASA+ group when compared to the ASA- group for anti-sperm secretory IgA (P < .001), anti-sperm monomeric IgA (P < .001), and anti-sperm IgG (P < .01). Seminal plasma levels were also raised in the ASA+ group, but only significantly so for monomeric IgA (P < .02). The performed ELISA has definite potential in research, especially with the use of monoclonal antibodies for the detection of anti-sperm secretory IgA, but cannot as yet be used as a prognostic predictor of fertility in the individual antibody-positive patient. Infertility specific antigens will have to be identified and isolated and subsequently used in the ELISA.Articl

The influence of solubilized porcine zona pellucida protein on the binding capacity of human spermatozoa

The acrosome reaction, sperm-zona pellucida binding, sperm-oolemma binding/fusion and subsequent fertilization are known to be influenced by homologous as well as heterologous follicular fluid and zona pellucida protein. In this study, the effect was investigated of different concentrations of solubilized porcine zona pellucida protein on the zona binding potential of human spermatozoa under hemizona assay conditions. Human spermatozoa incubated with 617 and 142 μg/ml porcine zona pellucida protein showed a statistically significant increase in zona binding when compared with control spermatozoa (106.5 ± 18.0 versus 60.9 ± 29.0, P 0.05). Control zona binding values for different experiments did not differ significantly (60.9 ± 29.0; 63.0 ± 25.5; and 66.7 ± 25.4, P > 0.6). In conclusion, it seems likely that a factor(s) present in the porcine zona pellucida might play a beneficial role during human spermo-ocyte binding. The results of the study might be used in future investigations to manipulate gamete interaction to such an extent that improved fertilization rates can be accomplished.Articl

The hemizona assay (HZA) as an experimental model to evaluate the inhibition of sperm binding to the murine zona pellucida by isolated zona pellucida protein

Compelling evidence has demonstrated that zona binding represents gamete recognition by sperm binding with high affinity and specificity to complex glycoproteins of the zona pellucida. In the present study we evaluated the hemizona assay (HZA) in the investigation of the interaction of mouse spermatozoa with unfertilized murine oocytes and hemizonae after exposure to solubilized murine zonae pellucidae proteins. The zonae pellucidae were isolated from ovarian tissue following described mincing techniques. The sperm binding characteristics of murine spermatozoa were studied by using SDS-PAGE isolated ZP2 (± 120 Kd) and ZP3 (± 83 Kd) components of the zona pellucida. Sperm receptor activity was examined in a competitive gamete binding fashion using the HZA as an indicator of sperm/zona interaction. The results illustrated that isolated, solubilized ZP2 and ZP3 glycoprotein moieties of the zona pellucida inhibited sperm binding to hemizonae and oocytes when compared to controls, and that the HZA can be utilized as an internally controlled homologous bioassay to evaluate the effects of zona pellucida proteins on tight binding of spermatozoa to the zona pellucida.Articl

A randomised placebo-controlled trial of the efficacy of beta-sitosterol and its glucoside as adjuvants in the treatment of pulmonary tuberculosis

OBJECTIVE: To evaluate the adjuvant effect of betasitosterol and its glucoside in the treatment of culture proven pulmonary tuberculosis (PTB). DESIGN: A blinded randomised placebo-controlled trial in culture proven drug sensitive PTB. Patients were hospitalised for the duration of treatment and evaluated at monthly intervals with regard to sputum culture positivity, chest radiography, weight gain, Mantoux test response, routine haematology and liver functions. STATISTICAL EVALUATION: General linear models for repeated measures (SAS GLM package) compared the interaction effects, group effects and time effects of findings in 19 patients receiving sitosterols with those in 18 patients receiving a placebo (talcum powder). Absolute values and change from baseline values were evaluated, although only the latter are reported. RESULTS: Weight gain was significantly greater in the sitosterol group (mean weight gain 8.9 kg) than the placebo group (mean gain 6.1 kg) (P = 0.0023 group effects; P = 0.0001 for time effects). Speed of achieving culture negativity, radiological improvement and induration on Mantoux testing was similar in the two groups. Change in lymphocyte counts from baseline was significantly higher in the sitosterol group (P = 0.0001 and P = 0.0001 for group and time effects) as was the increase in eosinophil counts (P = 0.0001 and P = 0.0137 for group and time effects). CONCLUSION: The study has shown significantly improved weight gain and higher lymphocyte and eosinophil counts in PTB patients receiving sitosterols in addition to an efficacious antituberculosis regimen. Sitosterols and their possible mode of action should now be evaluated in larger numbers of tuberculosis patients and in diseases with a similar immunopathogenesis.Articl