A phase I study of adenovirus-mediated wild-type p53 gene transfer in patients with advanced non-small cell lung cancer

Mutations of the tumor suppressor gene p53 are the most common genetic alterations observed in human cancer. Loss of wild-type p53 function impairs cell cycle arrest as well as repair mechanisms involved in response to DNA damage. Further, apoptotic pathways as induced by radio- or chemotherapy are also abrogated. Gene transfer of wild-type p53 was shown to reverse these deficiencies and to induce apoptosis in vitro and in preclinical in vivo tumor models. A phase I dose escalation study of a single intratumoral injection of a replication-defective adenoviral expression vector encoding wild-type p53 was carried out in patients with incurable non-small cell lung cancer. All patients enrolled had p53 protein overexpression as a marker of mutant p53 status in pretreatment tumor biopsies. Treatment was performed either by bronchoscopic intratumoral injection or by CT-guided percutaneous intratumoral injection of the vector solution. Fifteen patients were enrolled in two centers, and were treated at four different dose levels ranging from 10(7) to 10(10) PFU (7.5 x 10(9) to 7.5 x 10(12) particles). No clinically significant toxicity was observed. Successful transfer of wild-type p53 was achieved only with higher vector doses. Vector-specific wild-type p53 RNA sequences could be demonstrated in posttreatment biopsies of six patients. Transient local disease control by a single intratumoral injection of the vector solution was observed in four of those six successfully transduced patients. There was no evidence of clinical responses at untreated tumor sites. Wild-type p53 gene therapy by intratumoral injection of a replication-defective adenoviral expression vector is safe, feasible, and biologically effective in patients with advanced non-small cell lung cancer.     

Immunization status and sociodemographic characteristics: the mediating role of beliefs, attitudes, and perceived control

OBJECTIVES: This study examined how immunization-related beliefs, attitudes, and perceived control mediate up-to-date immunization among various sociodemographic groups. METHODS: Statewide estimates of immunization rates among children up to the age of 2 years were obtained via a multistage cluster sample. In-person interviews were conducted with 4832 parents. Information about immunization was obtained from official records or from health care providers. RESULTS: Differences in immunization among sociodemographic groups were mediated by beliefs about objective barriers to immunization, protection, medical contraindication, safety concerns, distrust, and natural immunity. Protection beliefs contributed to positive attitudes toward immunization; beliefs in natural immunity and safety concerns contributed to negative attitudes. Beliefs about objective barriers, distrust, safety concerns, and medical contraindications influenced perceived control over immunization. Positive attitudes and a strong sense of control contributed to higher immunization rates. CONCLUSION: These findings provide a basis for efficient educational campaigns by specifying which beliefs should be bolstered (because they facilitate proper immunization) and which should be targeted for change (because they hinder proper immunization) in various sociodemographic groups.     

Expansion of transplanted hepatocytes during liver regeneration

A method for volume selective proton spectroscopy is presented based on a multiecho sequence with short refocusing interval tcp. It is demonstrated, that by appropriate choice of tcp on the order of 4-6 ms, signals from overlapping multiplets like the glutamine and glutamate (Glu/Gln) resonances in spectra of the human brain are considerably increased compared with a conventional PRESS volume selection scheme. Thus proton spectra from J-coupled multiplet signals can be acquired with TE on the order of 20-30 ms avoiding the baseline problems arising at shorter echo times due to broad resonances. This allows to selectively acquire spectra from substances with longer T2 without the confounding effects from J-coupling occurring in conventional volume selection techniques.     

Risk factors for asthma in young adults. Spanish Group of the European Community Respiratory Health Survey

We assessed the association of asthma prevalence in young adults with susceptibility factors and environmental exposures, taking into account the age at asthma onset. A random sample of the general population, aged 20-44 yrs, in five areas of Spain (Albacete, Barcelona, Galdakao, Huelva, and Oviedo) was selected in the frame of the European Community Respiratory Health Survey (ECRHS). Overall, 2,646 subjects (response rate = 60.9%) responded to a respiratory questionnaire and 1,797 (41.4%) finalized a bronchial challenge. Atopy to perennial (odds ratio (OR) = 10.2, 95% confidence interval 4.2-25) and seasonal allergens (11.5, 4.6-28), parental asthma (4.5, 2.5-8.4), and birth order (OR for no older siblings in comparison to having more than two = 3.2, 1.2-9.1) were associated with current asthma whatever the age of asthma onset. Past asthma was associated to a lesser extent with atopy (OR around 3.5 to both perennial and seasonal allergens). Lower respiratory tract infections before the age of 5 yrs (LRTI), having had a pet in childhood, and being born in a younger cohort were associated with asthma starting before the age of 15 yrs, but not after. Male gender was more frequent in childhood asthma and female gender in adulthood. In addition to the known risk factors of asthma (atopy to perennial allergens, parental asthma) we provide evidence for an association of asthma (whatever the age of onset) with sensitization to seasonal allergens, and having less than three older siblings; and for an association of childhood asthma with lower respiratory tract infections.     

Etiology of genital ulcers and prevalence of human immunodeficiency virus coinfection in 10 US cities. The Genital Ulcer Disease Surveillance Group

To determine the etiology of genital ulcers and to assess the prevalence of human immunodeficiency virus (HIV) infection in ulcer patients in 10 US cities, ulcer and serum specimens were collected from approximately 50 ulcer patients at a sexually transmitted disease clinic in each city. Ulcer specimens were tested using a multiplex polymerase chain reaction assay to detect Haemophilus ducreyi, Treponema pallidum, and herpes simplex virus (HSV); sera were tested for antibody to HIV. H. ducreyi was detected in ulcer specimens from patients in Memphis (20% of specimens) and Chicago (12%). T. pallidum was detected in ulcer specimens from every city except Los Angeles (median, 9% of specimens; range, 0%-46%). HSV was detected in >/=50% of specimens from all cities except Memphis (42%). HIV seroprevalence in ulcer patients was 6% (range by city, 0%-18%). These data suggest that chancroid is prevalent in some US cities and that persons with genital ulcers should be a focus of HIV prevention activities.     

Cognitions, emotions, and behavior of patients with migraine when taking medication during an attack

Fifty-three patients with migraine, recruited from the Dutch Society of Migraine Patients and a general practice, were investigated regarding pain, moods, thoughts, and functioning during their most recent migraine attack, using a semistructured interview. Salient findings were: the high pain intensity the patients endured before they took analgesic medication, concerns about medication damaging their health, overoptimism regarding the effect of analgesic medication, and the relatively large proportion of patients (43%) who took medication primarily to be able to continue their activities. We recommend that future clinical trials on the effects of medication on migraine should not only include the measurement of pain during the attack, but also emotions, concerns about potential side effects and the ability to continue or resume work. Furthermore, it is important to provide patients with information about the side effects of medication and to apply cognitive-behavioral techniques for improvement of their mood during the attack.     

Biologically active Rep proteins of adeno-associated virus type 2 produced as fusion proteins in Escherichia coli

Four Rep proteins are encoded by the human parvovirus adeno-associated virus type 2 (AAV). The two largest proteins, Rep68 and Rep78, have been shown in vitro to perform several activities related to AAV DNA replication. The Rep78 and Rep68 proteins are likely to be involved in the targeted integration of the AAV DNA into human chromosome 19, and the full characterization of these proteins is important for exploiting this phenomenon for the use of AAV as a vector for gene therapy. To obtain sufficient quantities for facilitating the characterization of the biochemical properties of the Rep proteins, the AAV rep open reading frame was cloned and expressed in Escherichia coli as a fusion protein with maltose-binding protein (MBP). Recombinant MBP-Rep68 and MBP-Rep78 proteins displayed the following activities reported for wild-type Rep proteins when assayed in vitro: (i) binding to the AAV inverted terminal repeat (ITR), (ii) helicase activity, (iii) site-specific (terminal resolution site) endonuclease activity, (iv) binding to a sequence within the integration locus for AAV DNA on human chromosome 19, and (v) stimulation of radiolabeling of DNA containing the AAV ITR in a cell extract. These five activities have been described for wild-type Rep produced from mammalian cell extracts. Furthermore, we recharacterized the sequence requirements for Rep binding to the ITR and found that only the A and A' regions are necessary, not the hairpin form of the ITR.     

Amphetamines in hair by enzyme-linked immunosorbent assay

Human hair was collected from the occipital crown region of the head from several subjects; these hair samples were presumptively positive for amphetamines by a previously evaluated immunoassay. Hair was washed briefly with methanol to remove external contamination, then extracted with hot methanol for 2 h to recover the drugs. The extracts were evaporated to dryness, reconstituted in buffer, and analyzed using a new enzyme-linked immunosorbent assay (ELISA) technique adapted for the detection of amphetamines in hair. Gas chromatography-mass spectrometry was used as the reference technique. Cross-reactivity of several related compounds was evaluated by equating the inverse of the ligand concentration at 50% antibody binding to the affinity constant for each compound. The ratio of a compound's affinity constant to that for d-methamphetamine was used to derive percent crossreactivity. These experiments yielded values of 30.8% for d-amphetamine, 7.4% for I-methamphetamine, 4.3% for phentermine, 2.9% for I-amphetamine, and <1% for ephedrine, methylenedioxyamphetamine, and methylenedioxymethamphetamine. Cross-reactivity of unrelated compounds was found to be non-existent. The optimum cutoff concentration was determined by receiver operating characteristic curve analysis to be 300 pg/mg and the observed limit of detection was 60 pg/mg. Intra-assay precision at 300 pg/mg was 3.3% (coefficient of variation, CV), and the interassay CV was 10.5%. The sensitivity and specificity of the method were 83% and 92%, respectively.