Cardiomyocyte Differentiation from Mouse Embryonic Stem Cells by WNT Switch Method.

The differentiation of ESCs into cardiomyocytes in vitro is an excellent and reliable model system for studying normal cardiomyocyte development in mammals, modeling cardiac diseases, and for use in drug screening. Mouse ESC differentiation still provides relevant biological information about cardiac development. However, the current methods for efficiently differentiating ESCs into cardiomyocytes are limiting. Here, we describe the "WNT Switch" method to efficiently commit mouse ESCs into cardiomyocytes using the small molecule WNT signaling modulators CHIR99021 and XAV939 in vitro. This method significantly improves the yield of beating cardiomyocytes, reduces number of treatments, and is less laborious.

Dnmt3bas coordinates transcriptional induction and alternative exon inclusion to promote catalytically active Dnmt3b expression.

Embryonic expression of DNMT3B is critical for establishing de novo DNA methylation. This study uncovers the mechanism through which the promoter-associated long non-coding RNA (lncRNA) Dnmt3bas controls the induction and alternative splicing of Dnmt3b during embryonic stem cell (ESC) differentiation. Dnmt3bas recruits the PRC2 (polycomb repressive complex 2) at cis-regulatory elements of the Dnmt3b gene expressed at a basal level. Correspondingly, Dnmt3bas knockdown enhances Dnmt3b transcriptional induction, whereas overexpression of Dnmt3bas dampens it. Dnmt3b induction coincides with exon inclusion, switching the predominant isoform from the inactive Dnmt3b6 to the active Dnmt3b1. Intriguingly, overexpressing Dnmt3bas further enhances the Dnmt3b1:Dnmt3b6 ratio, attributed to its interaction with hnRNPL (heterogeneous nuclear ribonucleoprotein L), a splicing factor that promotes exon inclusion. Our data suggest that Dnmt3bas coordinates alternative splicing and transcriptional induction of Dnmt3b by facilitating the hnRNPL and RNA polymerase II (RNA Pol II) interaction at the Dnmt3b promoter. This dual mechanism precisely regulates the expression of catalytically active DNMT3B, ensuring fidelity and specificity of de novo DNA methylation.

Peri-operative complications of sphenopalatine artery ligation: a 10-year series from two secondary care centres.

BACKGROUND: There is currently limited evidence regarding the potential complications of sphenopalatine artery ligation. The post-operative outcomes at two secondary care centres over a 10-year period were reviewed. METHODS: A retrospective review was undertaken of patients undergoing emergency and elective sphenopalatine artery ligation between January 2011 and January 2021. Their demographics, peri-operative care and post-operative outcomes were recorded. The median follow-up time was 54 days (range, 0-2657 days). RESULTS: Ninety-one patients were included. Four patients (4.4 per cent) had a septal perforation at post-operative review. Nineteen patients (20.9 per cent) had post-operative bleeding that extended their in-patient stay, with five patients (5.5 per cent) requiring revision surgery. Pre-operative non-dissolvable nasal packing was used a median of 1 time (range, 0-8 times). CONCLUSION: Further research on outcomes of sphenopalatine artery ligation is needed. Pre-operative non-dissolvable nasal packing, concurrent septal surgical procedures, surgical techniques, and co-morbidities such as hypertension represent potential confounding factors that could not be further assessed in this small, retrospective study.

HIV-1 and the Mycobacterium tuberculosis granuloma: A systematic review and meta-analysis.

Infection with HIV-1 greatly increases the risk of active tuberculosis (TB). Although hypotheses suggest HIV-1 disrupts Mycobacterium tuberculosis (Mtb) granuloma function, few studies have examined this directly. The objective of this study was to determine what evidence exists about the effect HIV-1 co-infection has upon Mtb granulomas. A systematic search of PubMed, Web of Science, and Medline up to 20 March 2015 was conducted, to identify studies comparing Mtb-infected tissue from HIV-1 infected and uninfected persons, or HIV-1 infected persons with stratified peripheral CD4 T cell (pCD4) counts. We summarized findings that focused on how HIV-1 changes granuloma formation, bacterial presence, cellular composition, and cytokine production. Nineteen studies with a combined sample size of 899 persons were included. Although studies frequently were limited by variable or inadequately described definitions of outcomes and analytical methods, HIV-1 was found to be associated with increased bacillary load within Mtb-infected tissue. Reductions in pCD4 counts within co-infected persons associated with both poorer granuloma formation and higher bacterial load. The high degree of heterogeneity among studies combined with experimental limitations made it difficult to conclusively support previously published and prevalent hypotheses about HIV-1/Mtb co-infection granulomas. To elucidate the validity of these hypotheses we have described areas that can be improved in future studies in order to clarify the influence HIV-1 co-infection has upon the Mtb granuloma.

Sinus involvement in breast cancer: case report.

OBJECTIVE: To report a rare and unique presentation of metastatic breast cancer. METHOD: Case report and review of related literature. RESULTS: A 62-year-old female with metastatic breast cancer, who had been free from recurrence for 4 years, presented with diplopia secondary to lateral rectus palsy. This was due to a sphenoid sinus metastasis, which was eroding into her cavernous sinus, resulting in VIth cranial nerve neuropathy. CONCLUSION: All paranasal sinuses and the orbit are potential sites for metastases. Spread to the paranasal sinuses from breast cancer has been documented previously. However, we believe this to be the only reported case with lateral rectus palsy as a result of metastasis to the sphenoid sinus in which this was the only evidence of disseminated disease. Otolaryngology clinicians need to consider metastatic disease to the sinuses in patients with a history of neoplasia, but also recognise that tamoxifen treatment can itself cause visual disturbance.

Prospective, comparative, cohort study comparing the rhinogram, Sino-Nasal Outcome Test-20 and Heath-Related Quality of Life questionnaire.

BACKGROUND: Many outcome measures exist for rhinosinusitis. However, few are used in the clinical setting due to their long completion times. OBJECTIVE: To assess the validity, reliability and responsiveness of the rhinogram, compared with two validated rhinosinusitis outcome measures: the Sino-Nasal Outcome Test-20 and the Heath-Related Quality of Life questionnaire. METHODS: Fifty-one patients were entered into a prospective, comparative, cohort study using all three outcome measures one week pre-operatively and three months post-operatively. Outcome scores were then correlated using non-parametric Spearman's rank correlation and chi-square testing for the diagnostic criteria of all three outcome measures. RESULTS: Statistically significant correlations were found between all three outcome measures for all symptom scores, individually as well as combined (p < 0.01 for all calculations). Comparison of the diagnostic accuracy of the rhinogram, compared with the Sino-Nasal Outcome Test-20 and the Heath-Related Quality of Life questionnaire, showed statistical significance (p < 0.05; chi-square test). CONCLUSION: The rhinogram is a reliable, valid and responsive rhinosinusitis outcome measure which can assist patient diagnosis and management in the clinical setting. Due to its quick completion time, this outcome measure could be used in rhinology out-patient clinics.

Ecology of Increasing Diseases: Population Growth and Environmental Degradation.

The World Health Organization (WHO) and other organizations report that the prevalence of human diseases during the past decade is rapidly increasing. Population growth and the pollution of water, air, and soil are contributing to the increasing number of human diseases worldwide. Currently an estimated 40% of world deaths are due to environmental degradation. The ecology of increasing diseases has complex factors of environmental degradation, population growth, and the current malnutrition of about 3.7 billion people in the world.

Surgical management of soft tissue sarcoma in patients over 80 years.

AIMS: To report outcome on patients over 80years of age with soft tissue sarcoma (STS), with respect to surgical treatment, co-morbidity, complications and survival. METHODS: From a prospective database of 3400 patients with STS presenting over a 13-year period, all patients over 80years of age were identified and reviewed, with respect to tumour characteristics morbidity, mortality and outcome. RESULTS: 128 patients over 80years were treated for STS with 63 referred for treatment of primary disease, of whom 50 underwent resectional surgery. The remaining 65 patients were treated for recurrent or incompletely excised disease. Of the 50 patients treated primarily with surgery, 56% of tumours where high grade and 56% were greater than 10cm in diameter. The overall complication rate was 34%, with a 30-day mortality of 4%. Two- and 5-year survival rates were 56% and 46%, with a local recurrence rate of 22% at a mean follow-up of 22months. CONCLUSION: This patient group presented with poor prognosis tumours that were associated with poor outcomes in the medium to long term. Age need not be considered a contra-indication to radical surgery with curative intent.

Nasal deformity and interpersonal problems.

Several studies in facial plastic surgery have suggested that changes in body image that initiate a positive psychological cycle play an important role in a successful outcome. The change in the patient's self-perception leads to a sense of well-being, rather than there being a direct impact from the changed features. One area in which this change in self-perception may be manifest is in the development and maintenance of interpersonal relationships. Due to the impact of negative body image preoperatively, we hypothesized that patients with nasal deformity may have problems with their interpersonal relationships. This theory was tested using the IIP-32: a short version of the inventory of interpersonal problems. A prospective trial was undertaken of 27 rhinoplasty patients with nasal deformity and 16 septoplasty patients without nasal deformity, the latter group used as a control. The results show that the null hypothesis that the two groups are from the same population on all dimensions of the IIP-32 cannot be rejected (total mean score 0.753). From this data, it seems that patients with nasal deformity have no more difficulties in their interpersonal relationships and, as such, this particular issue cannot be used in support of rhinoplasty surgery.

Major ear surgery in a paediatric day care unit.

Abstract Traditionally major ear surgery in children has been regarded as an in-patient procedure. Evidence from the USA for adults, however, concludes that it is both safe and effective to perform many major ear procedures as day cases. We have been carrying out major ear operations on children as day cases routinely for six years in a dedicated children's day unit and examined our data to find out whether it was both safe and feasible to perform major ear surgery in children on a day-case basis. As our main outcome measure we used the rate of unplanned admissions. We found that the unplanned admission rate for surgery, excluding mastoid surgery, was 6.7 per cent and that procedures such as myringoplasty, ossiculoplasty, bilateral pinnaplasty, meatoplasty and tympanotomy with excision of cholesteatoma, were eminently suitable for day surgery.

Assessing psychological morbidity in patients with nasal deformity using the CORE questionnaire.

The aesthetic facial surgeon's evaluation of the patient seeking cosmetic surgery must be thorough. Those with emotional disturbances and overt psychiatric disorders are considered to be poor candidates for elective cosmetic surgery. In patients with nasal deformity, the prevalence of subclinical psychological morbidity and the role of cosmetic surgery in these circumstances are uncertain. This study investigates the use of the CORE questionnaire in assessing the psychological status of patients awaiting rhinoplasty or septorhinoplasty and compares this with the psychological status of patients awaiting septoplasty. A prospective trial of 27 rhinoplasty patients and 16 septoplasty patients revealed that in terms of life functioning, patients with nasal deformity have significantly higher psychological morbidity compared with patients without nasal deformity. However, there was no significant difference in global psychological status. In addition, female patients with nasal deformity are more distressed in terms of general wellbeing than male patients with nasal deformity and there is a tendency for higher global psychological impairment in women. These results may have implications in terms of health care rationing in patients with nasal deformity and preoperative psychological screening.

Topical bupivacaine in paediatric day-case tonsillectomy: a prospective randomized controlled trial.

Post-operative pain and delayed oral intake are common reasons for failure of discharge in day-case tonsillectomy. A double blind prospective randomized study was devised to investigate the effectiveness of topical bupivacaine in reducing post-operative pain in paediatric day-case tonsillectomy. Ninety-nine patients aged between three and 16 years were recruited into the study. One group received bupivacaine soaked swabs tightly packed in their tonsillar fossae while the control group received saline-soaked swabs. The bupivacaine group was found to drink (p < 0.001) and eat (p = 0.006) earlier than the control group. The pain scores at one (p < 0.001), three (p < 0.001) and six (p < 0.001) hours post-operatively were also found to be lower in the bupivacaine group than the control group. We conclude that topical bupivacaine has a role in facilitating recovery in day-case tonsillectomy in children.

Characterisation of [(125)I]-apamin binding sites in rat brain membranes with HE293 cells transfected with SK channel subtypes.

The pharmacology of [(125)I]-apamin binding sites was examined in rat cortical and hippocampal tissue and compared with membranes prepared from human embryonic kidney (HEK293) cells transfected with SK channel subtypes hSK1, rSK2 and rSK3. The K(D) of [(125)I]-apamin in rat cortex and hippocampus was similar to the apamin-sensitive subtypes, rSK2 and rSK3 (K(D) (pM): 6.4, 7.08, 6.56 and 8.94, respectively). In addition, [(125)I]-apamin had a K(D)=270.4pM for the putatively 'apamin-insensitive' hSK1. Apamin had about a three-fold higher affinity than [(125)I]-apamin in brain tissue and in the cells expressing the different SK channel subtypes. Pancuronium, bicuculline and d-tubocurarine displayed micromolar affinity for all five-membrane preparations, whereas dequalinium and gallamine appear to show some subtype selectivity. Tetraethylammonium (TEA) and 4-aminopyridine (4-AP) had millimolar affinity and linopirdine had no effect. In conclusion, the pharmacology of [(125)I]-apamin binding in the cortex and hippocampus was similar to that in the apamin-sensitive clones, rSK2 and rSK3. In addition, we demonstrated low affinity [(125)I]-apamin binding for hSK1 and identified compounds that show subtype selectivity. These data cast further doubt on the identification of SK1 as encoding for the K(+) channel responsible for the apamin-insensitive sAHP.

Functional laryngeal dyskinesia: an important cause of stridor.

Functional laryngeal dyskinesia is a disorder that presents with stridor. Failure to recognize its features can result in inappropriate investigation and treatment for a condition that has a psychogenic origin. The key feature in diagnosis is paradoxical vocal fold adduction on inspiration, as seen on fibre-optic nasendoscopy. This phenomenon together with the associated stridor may disappear after distraction techniques or when the patient is asleep. We present five such cases which presented to hospitals in south west London over a 12 month period.