Combined skin and muscle vaccination differentially impact the quality of effector T cell functions: the CUTHIVAC-001 randomized trial.

Targeting of different tissues via transcutaneous (TC), intradermal (ID) and intramuscular (IM) injection has the potential to tailor the immune response to DNA vaccination. In this Phase I randomised controlled clinical trial in HIV-1 negative volunteers we investigate whether the site and mode of DNA vaccination influences the quality of the cellular immune responses. We adopted a strategy of concurrent immunization combining IM injection with either ID or TC administration. As a third arm we assessed the response to IM injection administered with electroporation (EP). The DNA plasmid encoded a MultiHIV B clade fusion protein designed to induce cellular immunity. The vaccine and regimens were well tolerated. We observed differential shaping of vaccine induced virus-specific CD4 + and CD8 + cell-mediated immune responses. DNA given by IM + EP promoted strong IFN-γ responses and potent viral inhibition. ID + IM without EP resulted in a similar pattern of response but of lower magnitude. By contrast TC + IM (without EP) shifted responses towards a more Th-17 dominated phenotype, associated with mucosal and epidermal protection. Whilst preliminary, these results offer new perspectives for differential shaping of desired cellular immunity required to fight the wide range of complex and diverse infectious diseases and cancers.

What factors are important in smoking cessation and relapse in women from deprived communities? A qualitative study in Southeast England.

OBJECTIVES: Women are relatively more susceptible to smoking-related diseases and find it more difficult to quit; however, little research exists on factors associated with smoking cessation and relapse in women. We examined attitudes towards and perceptions of factors associated with smoking cessation and relapse in women from deprived communities. STUDY DESIGN: Qualitative interview study. METHODS: Participants included eleven women, smokers and ex-smokers, from disadvantaged communities in East Sussex, England, who had used the National Health Service (NHS) stop smoking service. Data were collected through a focus group and semi-structured interviews, and subjected to thematic analysis. RESULTS: Participants opined that it is more difficult for women to quit smoking than men. Women felt that postcessation weight gain was inevitable and acted as a barrier to quitting. Hormonal fluctuations during the menstrual cycle and greater levels of stress were perceived as obstacles to quitting and reasons for relapse. Conversely, the women cited effects of smoking on physical appearance, oral hygiene and guilt about exposing children to passive smoke as powerful motivators to quit; and highlighted the impact of public health campaigns that focused on these factors. Views diverged on whether quitting with someone close to you is a help or hindrance. Other themes including alcohol intake, daily routine and being in the presence of smokers emerged as situational triggers of relapse. CONCLUSIONS: Interventions that address women's concerns related to postcessation weight gain, hormonal fluctuations during the menstrual cycle and stress may aid with smoking cessation and reduce relapse. Public health campaigns should consider the impact of smoking on physical appearance and the effect of passive smoke on children.

Correlation between the clonogenic initial slope and the response of polykaryon-forming units: the behavior of strains defective in XRCC5 and ATM and the heritability of small variations in radioresponse.

The polykaryon-forming unit (PFU) assay measures the survival of multiple cycles of DNA synthesis after exposure to ionizing radiation, and it is known that there is a strong correlation between the slope of the PFU dose-response curve and the clonogenic initial slope. This suggests that DNA lesions expressed in clonogens are also important in PFU. Cells having a mutation in XRCC5 (also known as Ku80; strain xrs-6) and ATM (strain AT5BIVA) were hypersensitive in the PFU assay and in clonogens, while a strain of xrs-6 cells transfected with hamster wild-type XRCC5 cDNA displayed wild-type resistance in both assays. These data suggest that the DNA double-strand break (DSB) is an important lesion in PFU, although the relative radioresistance of PFU compared to clonogens indicates differential DSB toxicity. We propose that this results from the absence of cytokinesis-related loss of DNA fragments. Small variations in the radioresponse of PFU were observed between CHO K1 cell substrains, such that the xrs parental substrain RR-CHOK1 (carrying wild-type XRCC5) was more sensitive than an independent K1 substrain (E-CHOK1). Somatic hybridization showed that this variation is heritable and that the resistant E phenotype is dominant. In RR-CHOK1 cells there was a biphasic PFU radioresponse, which suggests that there may be transient expression at a locus selectively affecting PFU sensitivity.

Determination of radiosensitivity in established and primary squamous cell carcinoma cultures using the micronucleus assay.

In this study, the cytokinesis-block micronucleus assay (CBMN) was used to measure radiosensitivity in three established cell lines (SCC-61, V175 and V134) and 10 primary cell cultures of squamous cell carcinoma (SCC) of the head and neck. Assessment involved optimisation of the assay to determine cytochalasin-B (CB) concentration and sampling time postirradiation. A much closer correlation between dose-response data measured in the clonogenic and micronucleus assays was found when the micronucleus assay was performed under standardised conditions for each cell line (2 micrograms/ml CB: 48 h postirradiation) instead of predetermined optimised assay conditions. This indicates that, for these SCC cell lines, the CBMN assay may be able to predict in vitro radiosensitivity. To be of clinical use in predicting radiosensitivity, the CBMN assay also needs to be evaluated with primary cell cultures. In this study, no relationship between micronucleus frequency at 2 or 6 Gy and patient clinical outcome 12 months following surgery and radiotherapy was seen. Similarly, no association between patient outcome and tumour stage, nodal stage and histology was observed. These CBMN assay data from the primary cell cultures are presently inconclusive as a measure of patient tumour radiosensitivity.

Venereology as a specialty in Australia.

The development of venereology as a medical specialty began in 1979-1981 with the formation of multidisciplinary venereology societies in each state, followed by annual convention of an advocacy body, the National Venereology Council of Australia, which also included governmental representatives. In 1988 the Australian College of Venereologists was incorporated as a professional training body, and in 1992, the first Chair in Sexual Medicine was established by the Universities of Sydney and New South Wales. In parallel, the role of the nursing profession as active participants evolved dramatically: nurses work within the context of the health care team, with clinical, teaching, and outreach responsibilities, and by collaborating or initiating research. Sexual and Reproductive Health nursing is recognised as a specialist area, and the Australian Sexual Health Nurses Association (ASHNA) was inaugurated in 1991. Sexual Health Counselors come from a range of disciplines which represent the shift in focus from disease control to education and prevention, and which encompass sexual dysfunction, gender identity issues, sexual assault, and the empowerment of clients. Within the repertoire of many health care workers in sexually transmissable disease services are skills in the ¿new¿ public health (particularly health promotion), and an understanding of cultural influences on sexuality. ¿Sexual Health¿ has become the preferred name for such services.

The micronucleus assay: an evaluation of its use in determining radiosensitivity in vitro.

The cytokinesis-block micronucleus assay was used to measure radiosensitivity in vitro in a panel of seven cell lines. Six of these cell lines were used to study the major parameters of this assay. We observed varying sensitivities following cytochalasin-B exposure. Treatment with 1 microgram/ml cytochalasin-B for 24 h reduced cell survival in four of the six cell lines by > 60%. Cytochalasin-B concentration and post-irradiation culture time were both found to influence cell-response. In three cell lines (V39, V134 and HX142), a decrease in cytochalasin-B concentration (2-0.5 microgram/ml) resulted in an increase in the frequency of radiation-induced micronuclei per binucleate cell. In other cell lines, either the opposite (V7M, CHO-K1) or no effect (WiDr) was seen. A linear dose-response was observed between induced damage expressed as the frequency of micronuclei and radiation dose in all but one melanoma (V39) cell line. Evidence for radiation-induced division-delay, with the maximum frequency of binucleation in irradiated cultures occurring 24-48 h after that of controls, was only seen in two cell lines. Of particular note, and in contrast to some other published reports, was the lack of a general correlation between cell-response measured in the clonogenic and the cytokinesis-block micronucleus assays. Consideration of lethal lesions, determined from the clonogenic dose-response curve, with respect to micronucleus frequency showed a complex relationship, with one micronucleus per binucleate cell corresponding to a wide range of lethal lesions depending on the cell line.(ABSTRACT TRUNCATED AT 250 WORDS)

Exacerbation of bronchial asthma following treatment with amiodarone. Demonstration of an antiadrenergic effect in vitro.

We describe a patient with bronchial asthma whose respiratory symptoms worsened during treatment and rechallenge with amiodarone. Laboratory studies on cultured pulmonary cells demonstrated an anti-beta-adrenergic effect of amiodarone not due to inhibition of beta-adrenergic receptor binding. Amiodarone should be used with caution in patients with obstructive pulmonary disease.

Propionate assimilation in the flagellate Polytomella caeca. An inducible mitochondrial enzyme system.

1. The assimilation of propionate by Polytomella caeca involves the beta-oxidation of this fatty acid. 2. Propionate-grown cells immediately oxidize propionate, beta-hydroxypropionate, malonic semialdehyde and acetate; acetate-grown cells oxidize propionate rapidly only after a lag of 2hr., and this adaptation of resting cells to propionate involves the formation of the enzymes of beta-oxidation. 3. The beta-hydroxypropionate dehydrogenase and malonic semialdehyde dehydrogenase activities of both propionate-grown and propionate-adapted cells are partly located in mitochondrial fractions. 4. Mitochondria isolated from propionate-grown cells, and also those from acetate-grown cells fully adapted to propionate, oxidize succinate, alpha-oxoglutarate, beta-hydroxypropionate and malonic semialdehyde; oxidation of these substrates is tightly coupled to the phosphorylation of ADP. 5. Mitochondria from acetate-grown cells exhibit ADP-dependent oxidation of succinate and alpha-oxoglutarate, but do not oxidize beta-hydroxypropionate or malonic semialdehyde. Mitochondria isolated from acetate-grown cells adapted to propionate for 5hr. slowly oxidize beta-hydroxypropionate and malonic semialdehyde, but no tightly coupled phosphorylation is detectable. 6. Two of the inducible enzymes of propionate oxidation are located within the NAD-impermeable barrier and appear to be membrane-bound. 7. The formation of the inducible enzymes is inhibited by cycloheximide and actinomycin D, but not by chloramphenicol.

The regulation of propionate oxidation in Prototheca zopfii.

1. Whole cell suspensions of Prototheca zopfii grown on propionate oxidize propionate, acrylate, malonic semialdehyde and acetate immediately, whereas acetate-grown cells only oxidize acrylate or propionate rapidly after a lag of 20-30min. This adaptation to propionate is slowed down by 8-azaguanine or p-fluorophenylalanine, and is not influenced by adding an ammonium salt or an amino acid mixture. 2. The adaptation involves induction of the enzymes of beta-oxidation of propionate. 3. A small proportion (5-8%) of the activities of propionyl-CoA dehydrogenase, beta-hydroxypropionate dehydrogenase and malonic semialdehyde dehydrogenase are consistently associated with mitochondria isolated from propionate-grown cells. 4. Such mitochondria will oxidize propionyl-CoA, beta-hydroxypropionate and malonic semialdehyde, and the respiration rates with these substrates in the presence of inorganic phosphate are ADP-dependent. 5. Mitochondria from acetate-grown cells do not contain detectable activities of the enzymes of propionate oxidation.