Correction of SCID-X1 using an enhancerless Vav promoter.

The efficacy of gene therapy for the treatment of inherited immunodeficiency has been highlighted in recent clinical trials, although in some cases complicated by insertional mutagenesis and silencing of vector genomes through methylation. To minimize these effects, we have evaluated the use of regulatory elements that confer reliability of gene expression, but also lack potent indiscriminate enhancer activity. The Vav1 proximal promoter is particularly attractive in this regard and may be useful in situations where high-level or complex regulation of gene expression is not necessary. X-linked severe combined immunodeficiency (SCID-X1) is a good candidate for such an approach, particularly as there may be additional disease-related intrinsic risks of leukemogenesis, and where safety is therefore a paramount concern. We have tested whether lentiviral vectors expressing the common cytokine receptor gamma chain under the control of the proximal Vav1 gene promoter are effective for correction of signaling defects and the disease phenotype. Despite low-level gene expression, we observed near-complete restoration of cytokine-mediated STAT5 phosphorylation in a model cell line. Furthermore, at low vector copy number, highly effective T- and B-lymphocyte reconstitution was achieved in vivo in a murine model of SCID-X1, in both primary and secondary graft recipients. This vector configuration deserves further evaluation and consideration for future clinical trials.

A new PG13-based packaging cell line for stable production of clinical-grade self-inactivating gamma-retroviral vectors using targeted integration.

The clinical application of self-inactivating (SIN) retroviral vectors has been hampered by the lack of reliable and efficient vector production technologies. To enable production of SIN gamma-retroviral vectors from stable producer clones, a new PG13-based packaging cell, known as PG368, was developed. Viral vector expression constructs can be reliably inserted at a predefined genomic locus of PG368 packaging cells by an Flp-recombinase-mediated targeted cassette exchange (RMCE) reaction. A new, carefully designed vector-targeting construct, pEMTAR-1, eliminated the co-packaging of the selectable marker gene used for the identification of successful recombination at the predefined genomic locus and thus, improved the safety of the production system. Selected clones produced vector supernatants at consistent titers. The targeted insertion of therapeutically relevant SIN vectors for chronic granulomatous disease and X-linked severe combined immunodeficiency into PG368 cells results in stable titers within the range necessary for clinical application. The production of retroviral SIN vectors from stable clinical-grade producer cells is feasible and will contribute to the safe production and application of SIN gamma-retroviral vectors for clinical trials.

Long term neuropsychological outcome after head injury: relation to APOE genotype.

BACKGROUND: Existing evidence suggests that some patients who sustain a head injury suffer cognitive decline many years later, and that head injury and possession of the APOE epsilon 4 allele are each risk factors for Alzheimer's disease. OBJECTIVE: To determine whether late cognitive decline after head injury is more prevalent among carriers of APOE epsilon 4. METHODS: A database of head injured patients was used. Initial assessment was at the time of their injury, between 1968 and 1985, and outcome data at six months were available. Their ages at the time of injury ranged between 2 and 70 years. A cohort of 396 subjects was reassessed at a mean of 18 years later, with determination of APOE genotype and detailed neuropsychological testing. RESULTS: Judging by the Glasgow outcome scale, twice as many patients had deteriorated as improved between six months after injury and the late assessment; 22.2% of APOE epsilon 4 carriers had a good late outcome compared with 30.5% of non-carriers (95% confidence interval for the difference, -0.7% to 17.2%; p = 0.084). There were no clear differences between epsilon 4 carriers and non-carriers in detailed neuropsychological assessments. CONCLUSIONS: Although this study provides additional evidence that a late decline may occur after head injury, there was no clear relation to APOE genotype. Despite the follow up interval of 15 to 25 years, the cohort is still too young (mean age 42.1 years) to assess the risk of Alzheimer's disease.

The head injured adult--who cares?

A prospective observational survey of 2957 adults admitted to acute general hospitals in Glasgow after head injury, enabled the features of patients managed in the general hospital 48 hours after head injury to be compared with those transferred to the regional neurosurgical unit and those discharged from hospital.

Natural treatment of perennial allergic rhinitis.

Perennial allergic rhinitis is an IgE-mediated inflammatory disorder of the nasal mucosa characterized by paroxysms of sneezing, nasal congestion, pruritus, and rhinorrhea. The condition may be caused by certain environmental agents, food sensitivities, structural abnormalities, metabolic conditions, or synthetic drugs. Recent health impairment outcome studies on allergic rhinitis sufferers reveal a measurable decline in physical and mental health status and the inability to perform daily activities. Antihistamines, decongestants, anticholinergic agents, and corticosteroid drug therapy, alone or in combination, are typically used in the treatment of allergic rhinitis. Reported adverse side effects include sedation, impaired learning/memory, and cardiac arrhythmias. Therapeutic strategies should seek to decrease the morbidity already associated with this condition. Urtica dioica, bromelain, quercetin, N-acetylcysteine, and vitamin C are safe, natural therapies that may be used as primary therapy or in conjunction with conventional methods.

Disability in young people and adults one year after head injury: prospective cohort study.

OBJECTIVE: To determine the frequency of disability in young people and adults admitted to hospital with a head injury and to estimate the annual incidence in the community. DESIGN: Prospective, hospital based cohort study, with one year follow up of sample stratified by coma score. SETTING: Five acute hospitals in Glasgow. SUBJECTS: 2962 patients (aged 14 years or more) with head injury; 549 (71%) of the 769 patients selected for follow up participated. MAIN OUTCOME MEASURES: Glasgow outcome scale and problem orientated questionnaire. RESULTS: Survival with moderate or severe disability was common after mild head injury (47%, 95% confidence interval 42% to 52%) and similar to that after moderate (45%, 35% to 56%) or severe injury (48%, 36% to 60%). By extrapolation from the population identified (90% of whom had mild injuries), it was estimated that annually in Glasgow (population 909 498) 1400 young people and adults are still disabled one year after head injury. CONCLUSION: The incidence of disability in young people and adults admitted with a head injury is higher than expected. This reflects the high rate of sequelae previously unrecognised in the large number of patients admitted to hospital with an apparently mild head injury.

Clinical process learning to improve critical thinking.

A six-step process-learning strategy model serves as a framework for nursing students to critically analyze situations encountered during their clinical practice experience. Stephen Brookfield's four components of critical thinking and culturalization themes relate well to how nurses learn and experience critical thinking and serves as the model's organizing framework. This learning strategy has implications for all nurse educators because it incorporates the realities of nursing practice, merges nursing education with practice, involves students in affective, cognitive, and psychomotor domains of learning, and prepares graduates to function in dynamic and complex health care systems.

Evidence for a carbocation intermediate during conversion of bipinnatin-A and -C into irreversible inhibitors of nicotinic acetylcholine receptors.

The lophotoxins are naturally occurring antagonists of nicotinic acetylcholine receptors. These toxins are small diterpenes that irreversibly inhibit nicotinic receptors by specific covalent modification of Tyr190 in the alpha-subunits of the receptor. The naturally occurring lophotoxin analogs, bipinnatin-A and -C, are inactive protoxins. Activation of these toxins occurs spontaneously in buffer and involves replacement of the C2 acetate ester with a hydroxyl group. The mechanism involved in conversion of the inactive bipinnatins into their biologically active solvolysis products was investigated in this study. Solvolysis of bipinnatin-A in buffer containing [18O]water demonstrated that the C2 hydroxyl of the biologically active solvolysis product originated from the solvent. The rates of solvolysis of bipinnatins-A and -C were not affected by sodium azide. However, in the presence of azide, solvent products decreased and new azide-containing products appeared. Thus azide acted as a nucleophile after a rate-limiting step, such as the formation of a carbocation intermediate. The kaz/ks values for bipinnatin-A (2900 M-1) and bipinnatin-C (1450 M-1) suggest that the carbocation intermediates are relatively stable. Compounds capable of spontaneously generating carbocations may represent a novel new class of active-site-directed affinity reagents that can be applied to other receptors and enzymes.

Hospital clinical career advancement programs: comparing perceptions of nurse participants and nonparticipants.

The study reported in this article describes nurse participants and nonparticipants in hospital clinical career advancement programs (clinical ladders) and compares perceptions in relation to program intrinsic outcomes, needs, and criteria for level advancement. A stratified random sample consisted of 120 participants and 480 nonparticipants drawn from 1,769 clinical nurses employed full time at five regional general hospitals. The findings suggest that nurses' demographic characteristics do not influence program participation status and that clinical advancement programs have limited impact upon job enrichment designed to recruit and retain clinical nurses providing direct patient care. Program participants (106; 88.3 percent) differed significantly at the .05 level from nonparticipants (385; 80.2 percent) on each factor area measuring perceptions of clinical advancement programs.