Nuclear transporter Importin-13 plays a key role in the oxidative stress transcriptional response.

The importin superfamily member Importin-13 is a bidirectional nuclear transporter. To delineate its functional roles, we performed transcriptomic analysis on wild-type and Importin-13-knockout mouse embryonic stem cells, revealing enrichment of differentially expressed genes involved in stress responses and apoptosis regulation. De novo promoter motif analysis on 277 Importin-13-dependent genes responsive to oxidative stress revealed an enrichment of motifs aligned to consensus sites for the transcription factors specificity protein 1, SP1, or Kruppel like factor 4, KLF4. Analysis of embryonic stem cells subjected to oxidative stress revealed that Importin-13-knockout cells were more resistant, with knockdown of SP1 or KLF4 helping protect wild-type embryonic stem cells against stress-induced death. Importin-13 was revealed to bind to SP1 and KLF4 in a cellular context, with a key role in oxidative stress-dependent nuclear export of both transcription factors. The results are integral to understanding stress biology, highlighting the importance of Importin-13 in the stress response.

Reliability of repeated arm-crank cardiopulmonary exercise tests in patients with small abdominal aortic aneurysm.

Arm-crank ergometry may be useful in patients unable to pedal, for instance due to peripheral arterial disease. Twenty participants with small abdominal aortic aneurysm undertook two serial arm-crank tests and then a pedal test, four of whom had indeterminate anaerobic thresholds, precluding analysis. The mean (SD) peak arm and leg oxygen consumptions in 16 participants were 13.71 (2.62) ml.kg-1 .min-1 and 16.82 (4.44) ml.kg-1 .min-1 , with mean (SD) individual differences of 3.11 (2.48) ml.kg-1 .min-1 , p = 0.0001. The respective values at the anaerobic thresholds were 7.83 (1.58) ml O2 .kg-1 .min-1 and 10.09 (3.15) ml O2 .kg-1 .min-1 , with mean (SD) individual differences of 2.26 (2.34) ml O2 .kg-1 .min-1 , p = 0.0001. The correlation coefficients (95%CI) for peak oxygen consumption and anaerobic threshold were 0.88 (0.62-1.0) and 0.70 (0.32-1.0). There were no significant differences in serial arm-crank tests, with intracluster correlations (95%CI) of 0.87 (0.86-0.88) and 0.65 (0.61-0.69) for peak oxygen consumption and anaerobic threshold, respectively.

Postoperative pain following primary lower limb arthroplasty and enhanced recovery pathway.

INTRODUCTION: Enhanced recovery is a concept that has become increasingly popular for arthroplasty surgery over the last ten years. This study was designed to assess the analgesia requirements, pain levels and time to discharge for patients having primary arthroplasty in the enhanced recovery pathway. METHODS: A multidisciplinary prospective cohort study was carried out between January 2012 and March 2012. Data were collected for patients undergoing primary arthroplasty in one hospital during this time. Details of anaesthesia, local infiltration, additional medications and analgesia were recorded. A visual analogue scale pain score was obtained from each patient at time of mobilisation on days 0, 1, 2 and 3 postoperatively. RESULTS: Ninety-six patients were included in the study. Of these, 34 underwent total hip arthroplasty and 62 total knee arthroplasty (TKA). Pain was the greatest contributor for delayed discharge in TKA patients. The patients who had TKA and did not receive non-steroidal anti-inflammatory drugs (NSAIDs) had significantly higher pain scores (day 0, p<0.01; day 1, p<0.001; day 2, p<0.01) and significantly increased opiate demands compared with those patients who did receive NSAIDs. CONCLUSIONS: There are unacceptably high pain scores in patients undergoing TKA without the use of NSAIDs. There should be focused intervention with this group of patients to improve their pain scores and reduce their length of stay.

Nucleocytoplasmic shuttling of the Duchenne muscular dystrophy gene product dystrophin Dp71d is dependent on the importin α/ß and CRM1 nuclear transporters and microtubule motor dynein.

Even though the Duchenne muscular dystrophy (DMD) gene product Dystrophin Dp71d is involved in various key cellular processes through its role as a scaffold for structural and signalling proteins at the plasma membrane as well as the nuclear envelope, its subcellular trafficking is poorly understood. Here we map the nuclear import and export signals of Dp71d by truncation and point mutant analysis, showing for the first time that Dp71d shuttles between the nucleus and cytoplasm mediated by the conventional nuclear transporters, importin (IMP) α/ß and the exportin CRM1. Binding was confirmed in cells using pull-downs, while in vitro binding assays showed direct, high affinity (apparent dissociation coefficient of c. 0.25nM) binding of Dp71d to IMPα/ß. Interestingly, treatment of cells with the microtubule depolymerizing reagent nocodazole or the dynein inhibitor EHNA both decreased Dp71d nuclear localization, implying that Dp71d nuclear import may be facilitated by microtubules and the motor protein dynein. The role of Dp71d in the nucleus appears to relate in part to interaction with the nuclear envelope protein emerin, and maintenance of the integrity of the nuclear architecture. The clear implication is that Dp71d's previously unrecognised nuclear transport properties likely contribute to various, important physiological roles.

Quantitative analysis of localization and nuclear aggregate formation induced by GFP-lamin A mutant proteins in living HeLa cells.

Although A-type lamins are ubiquitously expressed, their role in the tissue-specificity of human laminopathies remains enigmatic. In this study, we generate a series of transfection constructs encoding missense lamin A mutant proteins fused to green fluorescent protein and investigate their subnuclear localization using quantitative live cell imaging. The mutant constructs used included the laminopathy-inducing lamin A rod domain mutants N195K, E358K, M371K, R386K, the tail domain mutants G465D, R482L, and R527P, and the Hutchinson-Gilford progeria syndrome-causing deletion mutant, progerin (LaA delta50). All mutant derivatives induced nuclear aggregates, except for progerin, which caused a more lobulated phenotype of the nucleus. Quantitative analysis revealed that the frequency of nuclear aggregate formation was significantly higher (two to four times) for the mutants compared to the wild type, although the level of lamin fusion proteins within nuclear aggregates was not. The distribution of endogenous A-type lamins was altered by overexpression of the lamin A mutants, coexpression experiments revealing that aberrant localization of the N195K and R386K mutants had no effect on the subnuclear distribution of histones H2A or H2B, or on nuclear accumulation of H2A overexpressed as a DsRed2 fusion protein. The GFP-lamin fusion protein-expressing constructs will have important applications in the future, enabling live cell imaging of nuclear processes involving lamins and how this may relate to the pathogenesis of laminopathies.

Granisetron (Kytril) suppresses methotrexate-induced nausea and vomiting among patients with inflammatory arthritis and is superior to prochlorperazine (Stemetil).

OBJECTIVE: Methotrexate (MTX) is an increasingly popular anti-rheumatic drug with its usefulness limited by toxicity, most commonly gastrointestinal (GI). The aim of the study was to study the effectiveness of the 5-HT3 receptor antagonist granisetron (GR) in the therapy of MTX-induced nausea. METHODS: A single-blind 8 week pilot study with random allocation to either GR 1 mg or prochlorperazine (Stemetil; PCh) 10 mg was undertaken in 13 patients who were taking or had taken MTX for either rheumatoid arthritis (10) or psoriatic arthritis (3). RESULTS: One in six patients treated with PCh completed the 8 week study compared to 7/7 treated with GR. After switching of symptomatic patients, 11 completed the study on GR and median improvement was by two grades (P < 0.001) with a significantly better visual analogue scale score for patient satisfaction compared to PCh. CONCLUSION: Treatment with GR may be useful in establishing and maintaining some patients on MTX where GI toxicity would have precluded such therapy.

Minocycline induced autoimmune hepatitis and systemic lupus erythematosus-like syndrome.

Monocycline is the most widely prescribed systemic antibiotic for acne largely because it needs to be given only once or twice a day and seems not to induce resistance. Up to April 1994 11 cases of minocycline induced systemic lupus erythematosus and 16 cases of hepatitis had been reported to the Committee on Safety of Medicines. An analysis of these cases together with seven other cases shows the severity of some of these reactions. Two patients died while taking the drug for acne and a further patient needed a liver transplant. Acne itself can induce arthritis and is often seen in association with autoimmine liver disease, but the clinical and biochemical resolution seen after withdrawal of the drug, despite deterioration of the acne, suggests a drug reaction. In five cases re-exposure led to recurrence. Because reactions may be severe early recognition is important to aid recovery and also to avoid invasive investigations and treatments such as corticosteroids and immunosuppresants. Safer alternatives should be considered for treating acne.

The use of topical 5% lignocaine ointment for the relief of pain associated with post-operative nasal packing.

Packing of the nasal cavity remains a common routine precautionary measure following septal surgery. The nasal pack and its removal 24 h later are often cited by patients as the most painful aspects of septal surgery. We present the results of a randomized, prospective controlled trial of the use of topical 5% lignocaine ointment as a method of pain relief following post-operative nasal packing. Post-operative pain as measured using a visual analogue scale at 3 h post-operatively was halved in patients receiving a lignocaine impregnated nasal pack compared with those having a standard vaseline gauze pack (P < 0.05). Pain scores at 6 h post-operatively and at pack removal were also reduced, but these failed to reach significance. No patients suffered reactionary haemorrhage. The use of topical lignocaine ointment is safe and may have a place in the relief of pain due to post-operative nasal packing.

Delayed linear growth in children with enterocystoplasties.

A series of 60 children whose urine was stored in pouches formed in whole or in part from bowel were reviewed to establish the effect on growth in height and weight. The mean age at operation was 7.8 years (range 0.9-14.1). Patients were included only if there were at least 2 measurements of both parameters pre-operatively and 2 post-operatively over a minimum follow-up of 2.5 years. Delayed growth in height alone occurred in 12 children (20%), with a mean loss of 25 percentile points (range 10-50). Delayed growth in height and weight occurred in 4. Accelerated growth in height alone, weight alone and in both occurred in 2, 7 and 7 respectively. There was no difference in a wide range of laboratory data between any of these groups. Although changes in growth rate (for better or worse) may occur for a variety of reasons after major surgery, delayed linear growth in isolation is an unusual and worrying finding that suggests a major alteration in bone development.

Blood and urine analysis in patients with intestinal bladders.

A group of 183 patients whose urine was stored in pouches formed either wholly or partly from bowel was reviewed to establish abnormal biochemical and haematological factors and to assess the risk of infection. Minimum follow-up was 1 year (range 1-28.5, mean 3 years 8 months) and the mean age of the patients at follow-up was 16.5 years (range 4-43). Pouches were augmentation and substitution cystoplasties and complete reconstructions with bowel. Bowel segments were ileum (55 patients), colon (100) and mixed (28). Anaemia was found in 15 patients, folate deficiency in 2 and folate excess in 2. Definite hyperchloraemic acidosis was found in 25 patients and borderline results in 40. Most of these patients had colon in the pouch but 5 definite and 13 borderline cases had ileum only. Troublesome urinary infections occurred in 27.9% of patients; 3 developed renal stones and 22 developed pouch/bladder stones. Stones were usually associated with infection and/or staples. Delayed linear growth occurred in 20% of children in spite of a normal increase in weight; there was no difference in any of the measurements between those who grew and those who did not.

Western equine encephalitis surveillance in Utah.

The history of WEE surveillance in Utah is reviewed, beginning with the 1933 outbreak involving 3,958 horses. The step by step formation of the Utah Mosquito Abatement Associations surveillance program from 1957 to the present is discussed. Results of an enlarged sentinel chicken flock surveillance program in Utah during 1983 (3 sero-conversions in September), 1984 and 1985 (no sero-conversion) show the lack of WEE activity in the surveillance area.