Cultures of Langerhans cells and co-culture with lymphoid cells: Relevance to toxicology and pharmacology

Langerhans cells are an essential component of cutaneous immunological defence mechanisms. They are mononuclear dendritic cells derived from bone marrow, which are present in the squamous epithelia, particularly skin. Langerhans cells show marked changes in phenotype and function when epidermal cell suspensions are prepared according to a trypsinization protocol and placed in culture for 1–3 days. They seem to mature and acquire most of the features of lymphoid dendritic cells, especially the interdigitating dendritic cells present in lymph nodes. This observation supports the assertion that resident Langerhans cells take up foreign antigens and then migrate through dermal lymphatics to local lymph nodes where they function as antigen-presenting cells. Thus, Langerhans cells play a pivotal role in the primary immune response and in the pathogenesis of skin diseases. In the past, most therapeutic treatment of skin diseases was achieved by administration of pharmacological agents by epicutaneous application. More recently, the determination of the toxicity of drugs and their pharmacological mechanisms has required the development of in vitro models to explore their potential effects at the cellular level. The effects of different agents (physical and chemical) on the density and function of fresh or cultured Langerhans cells are reviewed.     

Understanding cognitive development: automaticity and the early years child

In recent years a growing body of evidence has implicated deficits in the automaticity of fundamental facts such as word and number recognition in a range of disorders: including attention deficit hyperactivity disorder, dyslexia, apraxia and autism. Variously described as habits, fluency, chunking and over learning, automatic processes are best understood in terms of their distinctive properties. While typically identified as fast, parallel, attention-free processes, a commonly agreed definition of automaticity continues to elude theorists investigating this concept. Most theorists would, however, agree that since attentional resources are finite, automaticity of basic facts serves to free sufficient mental resources for a learner to focus their attention on the novel or more complex aspects of a task. Yet despite the importance of automaticity to the learner, the term remains largely unfamiliar to most educationalists and early years practitioners. In order to address this issue, the present paper seeks to review several influential theories of automaticity, to describe the problems associated with defining a process as automatic and to draw from relevant research to demonstrate how the early years environment can be organised to promote automaticity in the young learner.     

Effects of trypsin on the in situ identification of epidermal cell membrane antigens

In this work the role of trypsin in revealing epidermal cell surface antigens, with the use of immunological markers, was investigated. Two monoclonal antibodies (MCA) were used, the first: D47, belongs to the first cluster of differentiation and recognizes a membrane antigen of human thymocytes; the second HLA-ABC-m3, is an anti-HLA-B27 MCA. Preliminary treatment with various concentrations of trypsin was performed on frozen skin sections and followed by indirect immunofluorescence. D47 reacted with epidermal dendritic cells only after trypsin pretreatment of skin sections. In addition a mild preliminary trypsinization was shown to increase in situ immunoreactivity of MCA HLA-ABC-m3 with epidermal cells. Best results were obtained when trypsin concentrations ranging from 2.5 to 5 micrograms/ml were applied for 10 min at 37 degrees C. Preliminary trypsinization may be of interest for a better exposure of some surface antigens to immunohistochemical markers.     

Health status,work limitations,and return-to-work trajectories in injured workers with musculoskeletal disorders

BACKGROUND: The purpose of this study was to describe the health status and work limitations in injured workers with musculoskeletal disorders at 1 month post-injury, stratified by return-to-work status, and to document their return-to-work trajectories 6 months post-injury. METHODS: A sample of 632 workers with a back or upper extremity musculoskeletal disorder, who filed a Workplace Safety and Insurance Board lost-time claim injury, participated in this prospective study. Participants were assessed at baseline (1 month post-injury) and at 6 months follow-up. RESULTS: One month post-injury, poor physical health, high levels of depressive symptoms and high work limitations are prevalent in workers, including in those with a sustained first return to work. Workers with a sustained first return to work report a better health status and fewer work limitations than those who experienced a recurrence of work absence or who never returned to work. Six months post-injury, the rate of recurrence of work absence in the trajectories of injured workers who have made at least one return to work attempt is high (38%), including the rate for workers with an initial sustained first return to work (27%). CONCLUSIONS: There are return-to-work status specific health outcomes in injured workers. A sustained first return to work is not equivalent to a complete recovery from musculoskeletal disorders.     

High aggregate burden of somatic mtDNA point mutations in aging and Alzheimer's disease brain.

The mitochondrial theory of aging proposes that mitochondrial DNA (mtDNA) accumulates mutations with age, and that these mutations contribute to physiological decline in aging and degenerative diseases. Although a great deal of indirect evidence supports this hypothesis, the aggregate burden of mtDNA mutations, particularly point mutations, has not been systematically quantified in aging or neurodegenerative disorders. Therefore, we directly assessed the aggregate burden of brain mtDNA point mutations in 17 subjects with Alzheimer's disease (AD), 10 elderly control subjects and 14 younger control subjects, using a PCR-cloning-sequencing strategy. We found that brain mtDNA from elderly subjects had a higher aggregate burden of mutations than brain mtDNA from younger subjects. The average aggregate mutational burden in elderly subjects was 2 x 10(-4) mutations/bp. The bulk of these mutations were individually rare point mutations, 60% of which changed an amino acid. Control experiments ensure that these results were not due to artifacts arising from PCR error, mistaken identification of nuclear pseudogenes or ex vivo oxidation. Cytochrome oxidase activity correlated negatively with increasing mutational burden. These findings significantly bolster the mitochondrial theory of aging.     

Serological study of an allergic agranulocytosis due to noramidopyrine

An allergic agranulocytosis induced by amidopyrine and triggered by noramidopyrine was studied. Leucoagglutinating and leucocytotoxic antibody, active only in the presence of the drug, was demonstrated. The antibody was stable, giving a titre from 1·34 to 1·62 and was present in the IgM (19S globulin) and in the IgG (7S globulin) serum fractions. The site of drug fixation was studied by use of iodoantipyrine labelled with ¹³¹I; a stable fixation was demonstrated on to the IgM and IgG globulins. Special emphasis is given to cross-reaction with compounds related to amidopyrine.     

Somatic mitochondrial DNA mutations in cortex and substantia nigra in aging and Parkinson's disease

Oxidative damage to mitochondrial DNA (mtDNA) increases with age in the brain and can induce G:C to T:A and T:A to G:C point mutations. Though rare at any particular site, multiple somatic mtDNA mutations induced by oxidative damage or by other mechanisms may accumulate with age in the brain and thus could play a role in aging and neurodegenerative diseases. However, no prior study has quantified the total burden of mtDNA point mutation subtypes in the brain. Using a highly sensitive cloning and sequencing strategy, we find that the aggregate levels of G:C to T:A and T:A to G:C transversions and of all point mutations increase with age in the frontal cortex (FCtx). In the substantia nigra (SN), the aggregate levels of point mutations in young controls are similar to the levels in the SN or FCtx of elderly subjects. Extrapolation from our data suggests an average of 2.7 (FCtx) to 3.2 (SN) somatic point mutations per mitochondrial genome in elderly subjects. There were no significant differences between Parkinson's disease (PD) patients and age-matched controls in somatic mutation levels. These results indicate that individually rare mtDNA point mutations reach a high aggregate burden in FCtx and SN of elderly subjects.