Review: The role of mitochondria in the pathogenesis of amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by loss of upper and lower motor neurones leading to muscle weakness and paralysis. Despite recent advances in the genetics of ALS, the mechanisms underlying motor neurone degeneration are not fully understood. Mitochondria are known to be involved in the pathogenesis of ALS, principally through mitochondrial dysfunction, the generation of free radicals, and impaired calcium handling in ALS patients and models of disease. However, recent studies have highlighted the potential importance of altered mitochondrial morphology and defective axonal transport of mitochondria in ALS. Here, we review the evidence for mitochondrial involvement in ALS and discuss potential therapeutic strategies targeting mitochondria.

The effectiveness of an informal teaching programme for junior doctors identifying pleural effusions using ultrasound at the bedside.

OBJECTIVES: The use of bedside ultrasound to localise pleural effusions has gained in popularity in recent years. We investigated whether or not junior doctors could accurately identify thoracic anatomical structures and localise pleural effusions using bedside ultrasound. METHODS: Junior doctors were instructed by a consultant chest physician in the theory and practice of using ultrasound to localise pleural effusions, and then instructed in the method of inserting a Seldinger chest drain. Juniors were instructed to record a scan picture, including labelling of relevant structures. We then conducted a review of 52 images, noting indications, complications and the grade of doctor performing the scan. A consultant radiologist reviewed the scan pictures to confirm that the anatomy was correctly identified. RESULTS: Fifty out of 52 images (96%) were of diagnostic quality, with the anatomy correctly identified. The complication rate from chest drain insertion was 3.85%. CONCLUSIONS: With instruction, junior doctors can competently utilise basic chest ultrasound, obtain useful images, identify relevant thoracic anatomy and insert chest drains by the Seldinger technique with a low rate of complications. We suggest thoracic ultrasound should be more widely taught to junior doctors.

Treatment of antisocial personality disorder: Development of a practice focused framework.

There is little to no evidence of effective treatment methods for patients with an antisocial personality disorder (ASPD). One of the reasons could be the fact that they are often excluded from mental healthcare and thus from studies. A treatment framework based on 'state of the art' methods and best practices, offering guidelines on the treatment of ASP and possibilities for more systematical research, is urgently needed. This research involved a literature search and an international Delphi-study (N = 61 experts in research, management and clinical practice focused on ASPD). The results suggested important preconditions with regard to organization of care, healthcare workers and therapy. Conclusions are that there are many ways to coordinate effective treatment and management and work toward the increased availability of evidence based care for persons with ASPD.

Aberrant viruses in cells infected with murine sarcoma virus-feline leukemia virus.

This study describes an unusual type of virus seen when a feline leukemia virus (FeLV) pseudotype of murine sarcoma virus (MuSV) obtained by cocentrifugation procedures infected feline embryo cells (FEF) and two Crandell cat cell lines (CrFK1, CrFK2). When all three cell cultures were infected with MuSV-FeLV, only FEF and CrFK2 were transformed and only these showed normal and aberrant virus. The CrFK1 infected with MuSV-FeLV did not transform but did replicate normal type-C virus with a 50-A intermediate coat. The virus replicated in the two transformed lines showed three particles; a normal particle with a 50-A intermediate coat, a normal particle with a 100-A intermediate coat, and an aberrant particle with a 100-A intermediate coat.

Establishment of two canine sarcoma cell lines: productive infection with feline leukemia virus.

Two canine sarcoma cell lines (11028, 11031) were established in vitro and have been transferred 213 and 306 times, respectively, since 1970. These cell lines had a chromosome pattern consistent with their canine origin. Both cultures were infected with feline leukemia virus (FeLV), which caused a morphologic and karyotypic changes. The cells became rounded after infection with FeLV and both cultures showed the presence of a single, large, acrocentric chromosome considered to be a marker chromosome. All tumors were transplanted into newborn beagle pups, but only the 11028-FeLV formed metastatic tumors. No helper or focus-forming activity or virus particles were found in the uninfected cultures. Helper activity and virus were demonstrated in both sarcoma lines after infection with FeLV, though no focus-forming activity was noted. Helper activity of progeny virus could be assayed on either cat or dog embryo cells.

Growth of a heterologous tumor cell line in neonatal rats with graft-versus-host disease.

The ability of a hamster tumor cell line (T20) to grow and exhibit type H virus particles was significantly enhanced in neonatal DA rats with graft-versus-host disease (GVHD). Tumor growth peaked on days 4 and 7 in control littermates receiving adult syngeneic cells or no cells at birth, respectively, and then subsequently disappeared. However, tumor nodules in animals with GVHD became significantly larger than those in controls by day 7 and continued to grow until death. In addition, a marked plasma cell infiltration was noticed in such rapidly growing tumors from animals with GVHD only. In the light of previous studies, evidence is discussed for an environment within animals with GVHD conducive for tumor cell growth because of a depletion of T-cell areas within their tissues.

Infection and transformation of dog cells with a modified sarcoma virus.

Dog-embryo cell cultures were infected with a mixture of feline leukemia virus (FelLV) and a modified murine (Moloney) sarcoma virus MSV(FelLV). Morphological alteration of the cells was observed 3 days post infection. Virus progeny from the infected cultures increased 50- to 100-fold in foci of cellular alteration on dog cell cultures pretreated with diethylaminoethyl-dextran, but only twofold on cat embryo cultures similarly treated. The numbers of foci produced corresponded to a dual infection with the MSV(FelLV) and endogenous FelLV and could be increased by simultaneous infection with exogenous FelLV. After five serial passages in dog cell cultures, the virus mixture still had a superior focus-inducing capacity on cat cells as compared with dog cells. Electron microscopic examination of infected dog cultures showed typical C-type virions. FelLV alone propagated in dog cell cultures without inducing morphological alteration, and progeny virus, capable of promoting focus formation by MSV(FelLV), could be detected 48 hours post infection.

Cellular transmission of canine lymphoma and leukemia in beagles.

Thirty-four neonatal beagles were inoculated with cells obtained originally from 2 adult beagles, 1 with lymphocytic leukemia, the other with generalized malignant lymphoma. Most of the puppies were irradiated before inoculation; a few were inoculated in utero and were not irradiated. Of the 11 puppies inoculated with buffy coat cells of the blood from the leukemic donor, 2 developed leukemia. Of the 23 pups inoculated with cells of lymph nodes from the spontaneous lymphoma or positive passages, 13 developed lymphoma. Time of measurable onset of leukemia or lymphoma in the recipients varied from 12-60 days after inoculation. One passage was achieved with the leukemia and three serial passages with the lymphoma. A female karyotype was obtained from a neoplastic lymph node in a male recipient of the lymphoma. Since the donor was female, this suggests that the passages of the lymphoma were transplants.

Transformation of human embryo cells with the use of cell-free extracts of a human rhabdomyosarcoma cell line (HUS-2): brief communication.

Cell-free extracts of the human rhabdomyosarcoma cell line HUS-2 caused the transformation of human embryo fibroblasts. This transformation included morphologic alteration, karyotypic change, and an increase in culture longevity. With the use of sex markers, multiple karyotypes confirmed that the human embryo fibroblasts were transformed, and the use of cell-free material further suggested the presence of a transforming virus. RNA-dependent DNA polymerase activity in a particle with a specific gravity of 1.16 g/cm3 indicated the presence of an RNA type C virus. Evidence also suggested that the known mammalian type C viruses, routine cytopathic effect-inducing viruses, or mycoplasma were not the agents responsible for the transformation. That both the donor (HUS-2) and converted (HUE-T) cell lines cross-reacted with antisera prepared against HUE-T indicated a common antigen arising in the process of conversion of HUS-2 cells to HUE-T cells.

Epstein-Barr virus-specific cytotoxic T lymphocyte responses in the blood and tumor site of Hodgkin's disease patients: implications for a T-cell-based therapy.

Approximately 40% of Hodgkin's disease (HD) cases carry EBV in the malignant Hodgkin-Reed Sternberg (H-RS) cells, with expression of viral latent membrane proteins (LMPs) 1 and 2. These viral proteins are targets for CTLs in healthy EBV carriers, and their expression in EBV-associated HD raises the possibility of targeting them for a CTL-based immunotherapy. Here we characterize the CTL response to EBV latent antigens in both the blood and tumor-infiltrating lymphocytes of HD patients using two approaches: (a) in vitro reactivation of CTLs by stimulation with the autologous EBV-transformed lymphoblastoid cell line; and (b) an enzyme-linked immunospot assay to quantify frequencies of CTLs specific for known LMP1/2 epitopes. We detected EBV-specific CTLs in blood and biopsy samples from both EBV-negative and EBV-positive HD patients. However, as in healthy EBV carriers, LMP-specific CTL precursors occurred only at low frequency in the blood of HD patients, and with the exception of one EBV-negative HD case, were undetectable in the tumor. These data give rise to two considerations: (a) they may explain why EBV-positive tumor cells persist in the presence of an existing EBV-specific immune response; and (b) they provide a rationale for selectively boosting/eliciting LMP-specific CTL responses as a therapy for EBV-positive HD.

Qualitative research in healthcare: an introduction to grounded theory using thematic analysis.

In today's NHS, qualitative research is increasingly important as a method of assessing and improving quality of care. Grounded theory has developed as an analytical approach to qualitative data over the last 40 years. It is primarily an inductive process whereby theoretical insights are generated from data, in contrast to deductive research where theoretical hypotheses are tested via data collection. Grounded theory has been one of the main contributors to the acceptance of qualitative methods in a wide range of applied social sciences. The influence of grounded theory as an approach is, in part, based on its provision of an explicit framework for analysis and theory generation. Furthermore the stress upon grounding research in the reality of participants has also given it credence in healthcare research. As with all analytical approaches, grounded theory has drawbacks and limitations. It is important to have an understanding of these in order to assess the applicability of this approach to healthcare research. In this review we outline the principles of grounded theory, and focus on thematic analysis as the analytical approach used most frequently in grounded theory studies, with the aim of providing clinicians with the skills to critically review studies using this methodology.

Studies of a human mesothelioma.

Abdominal fluid from a patient with mesothelioma was cultured in vitro, and a cell line designated JMN was established. This cell line has undergone more than 100 passages. Electron microscopic, karyotypic, and heterotransplantation studies as well as the capability of collagen synthesis by this cell line indicated that it is of malignant mesothelial origin. The cell line has proved very useful for the isolation of human pathogenic viruses.

Mucolipidosis I.

A case of mucolipidosis I had clinical and histopathologic features that were a combination of changes found in both mucopolysaccharidoses and sphingolipidoses. Corneal clouding, spokelike cataracts, tortuous conjunctival and retinal vessels, and strabismus were clinical findings. Histopathologically, there were inclusion vacuoles similar to those seen in mucopolysaccharidoses; lamellar bodies typical of sphingolipidoses were rare. These vacuoles were seen in conjunctival and corneal epithelium and fibrocytes, conjunctival and retinal vessel endothelium, and all retinal cell layers.