BacMam production and crystal structure of nonglycosylated apo human furin at 1.89†Šresolution.

Furin, also called proprotein convertase subtilisin/kexin 3 (PCSK3), is a calcium-dependent serine endoprotease that processes a wide variety of proproteins involved in cell function and homeostasis. Dysregulation of furin has been implicated in numerous disease states, including cancer and fibrosis. Mammalian cell expression of the furin ectodomain typically produces a highly glycosylated, heterogeneous protein, which can make crystallographic studies difficult. Here, the expression and purification of nonglycosylated human furin using the BacMam technology and site-directed mutagenesis of the glycosylation sites is reported. Nonglycosylated furin produced using this system retains full proteolytic activity indistinguishable from that of the glycosylated protein. Importantly, the nonglycosylated furin protein reliably forms extremely durable apo crystals that diffract to high resolution. These crystals can be soaked with a wide variety of inhibitors to enable a structure-guided drug-discovery campaign.

Long-term follow-up of KTP laser turbinate reduction for the treatment of obstructive rhinopathy.

We investigated the subjective, long-term, patient-reported symptom control after endoscopic potassium titanyl phosphate (KTP) laser reduction of the inferior turbinates. Symptoms were listed preoperatively and patients were asked, via telephone interview, the status of their symptoms. Chi-square analysis was used to assess statistical significance of the surgical intervention. Thirty-six patients (male:female ratio: 5:4) aged 13 to 80 years (mean: 37.2) were available for follow-up. The mean follow-up time was 68.47 months (range: 44 to 92). The principal symptom patients experienced was nasal obstruction (94.4%); postoperative improvement in this symptom at 44 months and beyond was reported by 85.3% of patients (chi-square 16.94, p < 0.001). Other symptoms, such as rhinorrhea and sneezing, showed no significant improvement at long-term follow-up. This study adds evidence to the long-term improvements in obstructive nasal symptoms using KTP laser reduction of the turbinates.

The Weather and Ménière's Disease: A Longitudinal Analysis in the UK.

HYPOTHESIS: Changes in the weather influence symptom severity in Ménière's disease (MD). BACKGROUND: MD is an unpredictable condition that significantly impacts on quality of life. It is suggested that fluctuations in the weather, especially atmospheric pressure may influence the symptoms of MD. However, to date, limited research has investigated the impact of the weather on MD. METHODS: In a longitudinal study, a mobile phone application collected data from 397 individuals (277 females and 120 males with an average age of 50 yr) from the UK reporting consultant-diagnosed MD. Daily symptoms (vertigo, aural fullness, tinnitus, hearing loss, and attack prevalence) and GPS locations were collected; these data were linked with Met Office weather data (including atmospheric pressure, humidity, temperature, visibility, and wind speed). RESULTS: Symptom severity and attack prevalence were reduced on days when atmospheric pressure was higher. When atmospheric pressure was below 1,013 hectopascals, the risk of an attack was 1.30 (95% confidence interval: 1.10, 1.54); when the humidity was above 90%, the risk of an attack was 1.26 (95% confidence interval 1.06, 1.49). CONCLUSION: This study provides the strongest evidence to date that changes in atmospheric pressure and humidity are associated with symptom exacerbation in MD. Improving our understanding of the role of weather and other environmental triggers in Ménière's may reduce the uncertainty associated with living with this condition, significantly contributing to improved quality of life.

Mental Health and Subjective Well-being of Individuals With Ménière's: Cross-sectional Analysis in the UK Biobank.

HYPOTHESIS: Ménière's disease significantly impacts on an individual's mental health and subjective well-being (SWB). BACKGROUND: Ménière's disease is an unpredictable illness that impacts on mental health. The symptom triad of vertigo, tinnitus, and hearing loss all contribute to the disabling nature of the condition. To date, limited research has investigated the mental health and SWB impact of Ménière's disease. METHODS: We investigated the mental health and SWB of 1,376 Ménière's sufferers in the UK Biobank and compared this to over 500,000 controls. Participants in the Biobank were asked 38 questions pertaining to mental health and SWB. We utilized crude and adjusted linear and logistic regression to investigate the association between Ménière's and mental health and SWB. We also investigated how mental health and SWB might be related to length of diagnosis to see whether people might adapt to Ménière's over time. RESULTS: Ménière's was associated with increased frequency of depression, tiredness, tenseness, and unenthusiasm in the 2 weeks before recruitment. Ménière's was associated with longer periods of depression than controls (10.2 wk [95% CI: 5.2-15.2 wk]). Reduced health satisfaction was associated with Ménière's, but in other aspects of life (general happiness, work, family, friends, financial), individuals with Ménière's were as happy as controls. Mental health and SWB in individuals diagnosed for longer was better than in those who were recently diagnosed suggesting at least partial adaptation. DISCUSSION: This is the largest population study investigating the mental health impact of Ménière's. Our findings suggest that Ménière's adversely impacts on mental health, an individual's emotional state, and their life satisfaction. However, our findings raise the importance of supporting social relations for people with Ménière's and that although a cure is not currently available, we can still learn much about the adaptation strategies developed by long-term sufferers to help individuals with new diagnoses.

Patient satisfaction with postaural incision site.

Introduction. Controversy exists over the optimum incision placement when performing ear surgery via the postauricular approach. Little is known about the impact of incision placement on future comfort in wearing audio or visual aids or the effect on the minor auricular muscles cut in the approach. Objective. (1) To establish patient satisfaction with their postauricular surgical incision, and to establish the impact on comfort wearing hearing or visual aids. (2) To establish whether patients' voluntary ear movements were affected by surgery. Materials and Methods. In January 2014, questionnaires were sent to 81 patients who underwent mastoid surgery requiring a postauricular incision between January 2004 and December 2012. The incision placement was broadly the same for all patients as they were operated on by the same surgeon (or under his supervision). The incision is sited far posteriorly at the hairline. Results. 42 (52%) of the patients contacted responded. 80% of patients wearing glasses reported no discomfort or problems associated with their incision. 82% of patients who wear hearing aids were comfortable. Only 1 of the 5 patients who could move their ears preoperatively noticed a change afterwards. Conclusion. A hairline incision is well tolerated by most of the patients.

Endoscopically assisted branchial fistula excision.

We describe an endoscopically assisted dissection technique for cervical branchial fistulae that prevents the need for additional 'ladder' incisions in the neck. Two titanium clips are placed at the internal opening of the tract under endoscopic guidance through the initial skin incision. This technique is a useful adjunct to the usual technique if difficulties are experienced obtaining access higher in the neck. It enables an excellent view of surrounding structures.

Discovery of an inhibitor of insulin-like growth factor 1 receptor activation: implications for cellular potency and selectivity over insulin receptor.

Insulin-like growth factor 1 receptor (IGF-1R) is an attractive target for anti-cancer therapy due to its anti-apoptotic effect on tumor cells, but inhibition of insulin receptor (IR) may have undesired metabolic consequences. The primary sequences of the ATP substrate-binding sites of these receptors are identical and the crystal structures of the activated kinase domains are correspondingly similar. Thus, most small-molecule inhibitors described to date are equally potent against the activated kinase domains of IGF-1R and IR. In contrast, the non-phosphorylated kinase domains of these receptors have several structural features that may accommodate differences in binding affinity for kinase inhibitors. We used a cell-based assay measuring IGF-1R autophosphorylation as an inhibitor screen, and identified a potent purine derivative that is selective compared to IR. Surprisingly, the compound is a weak inhibitor of the activated IGF-1R tyrosine kinase domain. Biochemical and structural studies are presented that indicate the compound preferentially binds to the ATP site of non-phosphorylated IGF-1R compared to phosphorylated IGF-1R. The potential selectivity and potency advantages of this binding mode are discussed.