The Canadian Neuromuscular Disease Registry 2010-2019: A Decade of Facilitating Clinical Research Througha Nationwide, Pan-NeuromuscularDisease Registry.

We report the recruitment activities and outcomes of a multi-disease neuromuscular patient registry in Canada. The Canadian Neuromuscular Disease Registry (CNDR) registers individuals across Canada with a confirmed diagnosis of a neuromuscular disease. Diagnosis and contact information are collected across all diseases and detailed prospective data is collected for 5 specific diseases: Amyotrophic Lateral Sclerosis (ALS), Duchenne Muscular Dystrophy (DMD), Myotonic Dystrophy (DM), Limb Girdle Muscular Dystrophy (LGMD), and Spinal Muscular Atrophy (SMA). Since 2010, the CNDR has registered 4306 patients (1154 pediatric and 3148 adult) with 91 different neuromuscular diagnoses and has facilitated 125 projects (73 academic, 3 not-for-profit, 3 government, and 46 commercial) using registry data. In conclusion, the CNDR is an effective and productive pan-neuromuscular registry that has successfully facilitated a substantial number of studies over the past 10 years.

The Cancer, Lifestyle and Evaluation of Risk Study (CLEAR): Rationale and design of an unmatched "case-spouse control" study of over 10,000 participants in New South Wales, Australia.

INTRODUCTION: The New South Wales (NSW) Cancer, Lifestyle and Evaluation of Risk Study (CLEAR) is an open epidemiological bioresource, using an all cancer unmatched case-spouse control design. Participant characteristics and selected confirmed associations are compared to published estimates: current smoking and lung cancer; country of birth and melanoma; body mass index (BMI) and bowel cancer; and paternal history of prostate cancer and prostate cancer, to illustrate the validity of this design. MATERIAL AND METHODS: Cases are NSW residents, ≥18 years, with an incident cancer of any type. Controls are cancer-free spouses of cases. Participants complete a consent form, a questionnaire, and provide an optional blood sample. For analyses, odds ratios for males and females are calculated for cancers and exposures of interest, by sex-matching controls to cases. RESULTS: 10,816 participants (8569 cases, 2247 controls, 54% female) recruited to-date, median age: 61.6 y cases, 61.3 y controls. The top five cancer types are female breast (n=1691), prostate (n=1102), bowel (n=888), melanoma (n=608), and lung (n=265). Adjusted odds ratios (OR) were: 20.65 (95% CI: 13.25-32.19) for lung cancer in current versus never smokers; 1.16 (1.05-1.28) for bowel cancer per 5 kg/m(2) increment in BMI; 1.41 (1.01-1.96) for melanoma in Australian-born compared to those born in UK/Ireland; and 2.47 (1.82-3.37) for prostate cancer in men with versus without a paternal history of prostate cancer. DISCUSSION: This study design, where controls are the spouses of cases diagnosed with a variety of cancers and which are analysed unmatched, avoids potential biases due to overmatching, considered problematic in standard case-spouse control studies, and illustrates that risk estimates analysed are consistent with the published literature. CLEAR methodology provides a practical design to advance local knowledge on the causes of various leading and emerging cancers.

Responsiveness of the EQ-5D health-related quality-of-life instrument in assessing low back pain.

OBJECTIVE: To compare the responsiveness of the EuroQol five-dimensional questionnaire (EQ-5D) generic quality-of-life instrument with that of specific instruments-the Brief Pain Inventory (BPI) and the Oswestry Disability Index (ODI)-in assessing low back pain. METHODS: Data were obtained from a group of patients receiving epidural steroid injections. We assessed responsiveness by using correlation, by estimating standardized response means, by receiver operating characteristic curve analysis, and by comparing the minimum clinically important differences peculiar to each of the instruments. RESULTS: ODI, BPI, and EQ-5D index scores, and changes in scores, were found to be correlated. Estimated standardized response means and receiver operating characteristic curve analysis suggested lower responsiveness for the EQ-5D index score. Clinically significant categories of mild, moderate, and severe BPI pain intensity translated into progressively and significantly lower mean EQ-5D index scores. An increase or a decrease in severity level reported on any of the five EQ-5D dimensions was associated with significant changes (with appropriate signs) in the condition-specific scores. No change in severity in any EQ-5D dimension was associated with no change in the specific scores. Significant changes in the EQ-5D index scores were associated with clinically important changes in the ODI and BPI scores. Correlation between index scores and responses on EQ-5D's visual analogue scale was only moderate. CONCLUSIONS: The EQ-5D index is less responsive than instruments specific to pain measurement, although it is capable of indicating clinically important changes. The lower responsiveness arises from EQ-5D's more limited gradation of severity and its multidimensionality.

A pilot study of the dose-response of caudal methylprednisolone with levobupivacaine in chronic lower back pain.

The question as to what constitutes the ideal epidural steroid injection remains unresolved. We performed a prospective, randomised, double-blind, AB/BA 2 × 2 crossover study of caudal 40 vs 80 mg methylprednisolone acetate (in 20 ml levobupivacaine 0.125%) in outpatients with chronic low back pain. Data from 33 participants were analysed. The Oswestry Disability Index improved in both dose groups over time following injection. However, a statistically significant improvement was only observed in the 40 mg methylprednisolone acetate group (40 mg: p < 0.001; 80 mg: p = 0.33). There was no statistically significant difference between the dose groups in change in the Oswestry Disability Index with respect to time. Methylprednisolone acetate 40 mg appears to be as effective as 80 mg in improving disability associated with chronic low back pain, and should be considered in preference to the 80 mg dose for outpatients with chronic low back pain attending for repeat caudal steroid injection.

In vitro maintenance of Leishmania amastigotes directly from lesions: advantages and limitations.

Leishmania amazonensis (MHOM/BR/77/LTB0016) amastigotes were obtained from mouse cutaneous lesions and maintained in vitro for 48 hr at pH 4.6, 33 C. These organisms were reproducing, capable of transformation to promastigotes, and did not display the promastigote-specific antigen, GP46. In contrast, 97% of the organisms maintained for 24 hr at 31 C, pH 7.3, were positive for GP46. Thus, short-term cultivation of this L. amazonensis strain under appropriate conditions can provide a high yield of amastigotes for various in vivo and in vitro studies. However, the possible interference of host immunoglobin on the surface of these amastigotes needs to be considered because fluorescent-labeled anti-mouse immunoglobulin was detected on 16% of lesion-derived amastigotes even after 114 hr of cultivation.

Leishmania amazonensis: cultivation and characterization of axenic amastigote-like organisms.

Extracellular amastigote-like forms of Leishmania amazonensis can be maintained in axenic culture at 32 degrees C, pH 4.6, with a generation time of approximately 17 hr. This species of Leishmania is of particular interest since it has been associated with cutaneous, diffuse cutaneous, and mucocutaneous forms of the disease. Immunofluorescence, Western and Northern blot analyses, and immunoprecipitation have been used to estimate the expression levels of amastigote or promastigote antigens in axenically cultured amastigotes. In these analyses, monoclonal antibodies (mAbs) specific for either the amastigote (A-1, A-2, P-2, P-4, P-5, P-8) or promastigote (M-2, P-9, and F-4) and a DNA probe that was specific for the amastigote gene encoding the protein reactive with mAb P-4 have been employed. The amastigote-like organisms were infective for peritoneal and J774.G8 macrophages and BALB/c mice. While amastigote-like forms maintained at pH 4.6, 32 degrees C transformed to promastigotes when transferred to pH 7.3, 24 degrees C, transformation of promastigotes to amastigote-like organisms required a period of growth at pH 4.6 24 degrees C prior to transfer to 32 degrees C. This is the first report of the axenic cultivation of L. amazonensis amastigote-like organisms. This species grows in continual culture at a lower pH than any other species characterized to date. These organisms will prove useful in further studies of the biochemistry, immunology, developmental biology, and molecular biology of this parasite.

Developmental life cycle of Leishmania--cultivation and characterization of cultured extracellular amastigotes.

The biochemistry and immunology of Leishmania promastigotes has been extensively studied; this is due primarily to the facility with which this stage, in contrast to the amastigotes stage, can be maintained in axenic culture. Several attempts to axenically culture lines of Leishmania amastigotes have been reported in the literature. This paper summarizes methods of adaptation (low pH, elevated temperature and culture medium) and characterization of several axenic lines of Leishmania amastigotes. Based on morphological, biological, immunological and biochemical evidence, these organisms appear to resemble amastigotes from infected macrophages or tissue. The axenically cultured amastigotes appear to be distinct from shocked (heat, serum deprivation, stressed) Leishmania promastigotes in the plethora of proteins synthesized, growth (multiplication) in culture, and developmental regulation observed. These data suggest that Leishmania organisms have a significant developmental response to certain signals (pH, temperature) mimicking their in vivo macrophage milieu. The response to other environmental parameters characteristic of the host-macrophage remain to be determined. These axenically cultured amastigotes should be of interest for further immunological, biochemical and developmental investigations of the disease-maintaining stage of this parasite.

Leishmania enriettii amastigotes: demonstration of fibrillar surface.

An extensive fibrillar surface was identified in situ by transmission electron microscopy on the amastigote stage of the macrophage intracellular parasite, Leishmania enriettii. Attachment to the amastigote surface of 20-160-nm fibrils was clearly evident on organisms that had been isolated from guinea pigs. Surface fibrils were not detected after in vitro transformation of the amastigote to the promastigote. Incubation of amastigotes in heat-inactivated guinea pig serum containing a high titer of antibodies to L. enriettii followed by exposure to ferritin-conjugated antibody to guinea pig IgG resulted in labeling of surface fibrils, a result indicating that these fibrils are of parasite origin. Shedding of labeled fibrils was also evident.

In vivo assay of viability of amastigotes of Leishmania donovani.

Following intravenous (i.v.) injection of formalin-killed or ultraviolet-killed or ultraviolet-irradiated amastigotes of Leishmania donovani, the parasites could be identified in mouse liver at 5 min, but most amastigotes were digested by 3 hr or between 24 and 72 hr, respectively. An in vivo assay of viability, based on these observations, suggests differences in the viability of hamster amastigote populations. The identification of dead and/or dying amastigotes in the liver 24 hr after i.v. injection suggests that enumeration of amastigotes from Giemsa-stained slide preparations may include varying numbers of nonviable organisms.