Microbicides for HIV/AIDS. 3. Observation of apparent dynamic protonation and deprotonization in CD4+ T-cell model systems.

New measurements of the electrophoretic mobility of T-cell model systems have been carried out and analyzed to obtain the dynamic variation in mobility in small titration increments during separate upscale and downscale sweeps in pH. We demonstrate that a plot of plambda vs p[NaCl] has been found essential in evaluating the consistency of electrophoretic mobility measurements at different (1:1) electrolyte concentrations and show, for the first time, that electrophoretic mobility measurements as a function of pH can reflect different rates of the respective ionization and association that occur in the surface functional groups as a consequence of the different changes in the hydration-dehydration reactions involved. Differences found between the upscale and downscale sweeps suggest that it is easier to protonate a protein cell surface than to deprotonate it. The effect is most pronounced at the highest salt concentration (similar to that which exists for the cells in their native state) and becomes less pronounced as the salt concentration is lowered. The effect is interpreted as a result of the different changes in the state of hydration as a proton moves from the bulk through the double layer to a surface group and the reverse. The effect occurs with both replicating and activated T-cells. This latter result may be of biological significance and particularly relevant to HIV-1 infection, since during male-to-female transmission, the environment where most infections occur supports this protonation effect.

Safety and availability of dapivirine (TMC120) delivered from an intravaginal ring.

Vaginal delivery of 200 mg or 25 mg dapivirine from intravaginal rings (IVRs) was evaluated over a 7-day period in two phase 1 safety trials (IPM001 and IPM008, respectively) in a total of 25 healthy women 19 to 46 years of age. The IVR was generally safe and well tolerated with similar adverse events observed in the placebo and dapivirine groups. Across both studies, dapivirine concentrations in vaginal fluids measured at the introitus, cervix, and ring area were within the mean range of 0.7-7.1 microg/ml. Mean dapivirine concentrations in vaginal and cervical tissues on day 7 were 0.3-0.7 microg/g in IPM001 and 1.5-3.5 microg/g in IPM008. Mean plasma concentrations of dapivirine were <50 pg/ml. Dapivirine from both IVRs was successfully distributed throughout the lower genital tract at concentrations >1000x the EC(50) against wild-type HIV-1 (LAI) in MT4 cells suggesting that IVR delivery of microbicides is a viable option meriting further study.

Microbicides for HIV/AIDS. 2. Electrophoretic fingerprinting of CD4+ T-cell model systems.

New measurements of the dependence of the surface charge on the pH and electrolyte concentration for three living human white blood cell lines that are the principal targets of the HIV-1 virus are reported. Comparison of the electrophoretic fingerprint (EF) pattern, especially the line of zero mobility, with that of reference colloids establishes the separate individual identities and shows that all three exhibit a zwitterionic surface. With the EF results as a guide, preliminary biological infectivity measurements showed that small polyvalent cations modulate the negative charge on the T-cell surface in a way that strongly affects the infection kinetics. H9 cells were exposed to an infectious virus (X4), and the data showed that HIV interaction with target cells is enhanced by physiological fluids. The nondestructive methodology described is generally applicable to characterization of the surface charge and determination of the colloidal stability of any aqueous charged colloidal system without reference to any model of the double layer.

Microbicides for HIV/AIDS. 1. Electrophoretic fingerprinting the H9 cell model system.

An electrophoretic fingerprint of a CD4+ T-cell (H9) has been produced for the first time. Samples were taken from three separate cultures prepared at different times to obtain a general characterization of the cells. The availability of commercial instrumentation equipped with an auto-titrator has made possible the application of both the 2-dimensional and 3-dimensional representation of electrophoretic fingerprinting. The 2-dimensional treatment has been used to assess the reliability of the data and has detected hysteresis as a possible second-order effect. The 3-dimensional representation has been used to explore the data needed for a reliable overall pattern that characterizes the conditions of pH and conductivity required for an effective microbicide. The dome negative maximum in the electrophoretic fingerprint at high pH, along with the line of zero mobility (LZM) and a dome positive maximum at low pH, are interpreted as evidence for surface carboxyl groups prominent in the alkaline regime and surface amino groups prominent in the acid regime, suggesting that the H9 cell surface is zwitterionic. This has important implications as to the choice and design of microbicide actives.

Microfine zinc oxide is a superior sunscreen ingredient to microfine titanium dioxide.

BACKGROUND: Microfine zinc oxide and microfine titanium dioxide are particulate sunscreen ingredients that absorb broad-spectrum ultraviolet (UV) irradiation. OBJECTIVE: We compare microfine zinc oxide and microfine titanium dioxide for their abilities to attenuate UVA radiation and their relative whiteness in cosmetic formulations. METHODS: UVA attenuation was measured by diffuse reflectance spectroscopy on normal human skin in vivo. Whiteness was determined by reflectance density of dried coatings on a black background of the two particulates at varying concentrations. RESULTS: Microfine zinc oxide demonstrates superior protection compared to microfine titanium dioxide in the UV spectrum between 340 and 380 nm. Microfine zinc oxide is less white than titanium dioxide at all concentrations. CONCLUSION: Microfine zinc oxide is superior to microfine titanium dioxide as a sunscreen ingredient. It is more protective against long-wave UVA and is less white at a given concentration.

Microfine zinc oxide (Z-cote) as a photostable UVA/UVB sunblock agent.

BACKGROUND: Microfine zinc oxide (Z-Cote) is used as a transparent broad-spectrum sunblock to attenuate UV radiation (UVR), including UVA I (340-400 nm). OBJECTIVE: Our purpose was to assess the suitability of microfine zinc oxide as a broad-spectrum photoprotective agent by examining those properties generally considered important in sunscreens: attenuation spectrum, sun protection factor (SPF) contribution, photostability, and photoreactivity. METHODS: Attenuation spectrum was assessed by means of standard spectrophotometric methods. SPF contribution was evaluated according to Food and Drug Administration standards. Photostability was measured in vitro by assessing SPF before and after various doses of UVR. Photoreactivity was evaluated by subjecting a microfine zinc oxide/organic sunscreen formulation to escalating doses of UVR and determining the percentage of organic sunscreen remaining. RESULTS: Microfine zinc oxide attenuates throughout the UVR spectrum, including UVA I. It is photostable and does not react with organic sunscreens under irradiation. CONCLUSION: Microfine zinc oxide is an effective and safe sunblock that provides broad-spectrum UV protection, including protection from long-wavelength UVA.

A review of sunscreen safety and efficacy.

The use of sunscreen products has been advocated by many health care practitioners as a means to reduce skin damage produced by ultraviolet radiation (UVR) from sunlight. There is a need to better understand the efficacy and safety of sunscreen products given this ongoing campaign encouraging their use. The approach used to establish sunscreen efficacy, sun protection factor (SPF), is a useful assessment of primarily UVB (290-320 nm) filters. The SPF test, however, does not adequately assess the complete photoprotective profile of sunscreens specifically against long wavelength UVAI (340-400 nm). Moreover, to date, there is no singular, agreed upon method for evaluating UVA efficacy despite the immediate and seemingly urgent consumer need to develop sunscreen products that provide broad-spectrum UVB and UVA photoprotection. With regard to the safety of UVB and UVA filters, the current list of commonly used organic and inorganic sunscreens has favorable toxicological profiles based on acute, subchronic and chronic animal or human studies. Further, in most studies, sunscreens have been shown to prevent the damaging effects of UVR exposure. Thus, based on this review of currently available data, it is concluded that sunscreen ingredients or products do not pose a human health concern. Further, the regular use of appropriate broad-spectrum sunscreen products could have a significant and favorable impact on public health as part of an overall strategy to reduce UVR exposure.