Automated real-time detection of drug-resistant Mycobacterium tuberculosis on a lab-on-a-disc by Recombinase Polymerase Amplification.

With the emergence of multi- and extensive-drug (MDR/XDR) resistant Mycobacterium tuberculosis (M. tb), tuberculosis (TB) persists as one of the world's leading causes of death. Recently, isothermal DNA amplification methods received much attention due to their ease of translation onto portable point-of-care (POC) devices for TB diagnosis. In this study, we aimed to devise a simple yet robust detection method for M. tb. Amongst the numerous up-and-coming isothermal techniques, Recombinase Polymerase Amplification (RPA) was chosen for a real-time detection of TB with or without MDR. In our platform, real-time RPA (RT-RPA) was integrated on a lab-on-a-disc (LOAD) with on-board power to maintain temperature for DNA amplification. Sputa collected from healthy volunteers were spiked with respective target M. tb samples for testing. A limit of detection of 102 colony-forming unit per millilitre in 15 min was achieved, making early detection and differentiation of M. tb strains highly feasible in extreme POC settings. Our RT-RPA LOAD platform has also been successfully applied in the differentiation of MDR-TB from H37Ra, an attenuated TB strain. In summary, a quantitative RT-RPA on LOAD assay with a high level of sensitivity was developed as a foundation for further developments in medical bedside and POC diagnostics.

Association between dietary intake and 'school-valued' outcomes: a scoping review.

Approximately one in four Australian children aged 5-17 years are overweight or obese. Most of the health effects of overweight and obesity in childhood do not eventuate until into adulthood; therefore, motivation for children to have a healthy diet may be low. This scoping review examined the literature for associations between diet quality in 5-18 year olds and 'school-valued' outcomes including student attendance, academic performance, behaviour at school and mental health. A literature search for studies that assessed dietary intake and at least one 'school-valued' outcome in schoolchildren, in highly developed countries was conducted. After applying selection criteria, 35 studies were included examining academic performance (46%), behaviour (11%), mental health (31%) and 11% examining two of these outcomes each. No relevant studies addressed attendance. In general, dietary factors including consumption of fruit and vegetables, discretionary foods and/or beverages, or overall diet quality, were suggested to be correlates of the 'school-valued' outcomes. However, the evidence is not comprehensive. This review elucidates the extent and nature of available literature, and provides a basis for future research where the potential benefits of diet on 'school-valued' outcomes can be thoroughly explored.

Severity of airflow limitation, co-morbidities and management of chronic obstructive pulmonary disease patients acutely admitted to hospital.

OBJECTIVE: To assess the disease spectrum, severity of airflow limitation, admission pattern, co-morbidities, and management of patients admitted for acute exacerbations of chronic obstructive pulmonary disease. DESIGN: Case series. SETTING: An acute regional hospital in Hong Kong. PATIENTS: Adult subjects admitted during January 2010 to December 2010 with the principal discharge diagnosis of chronic obstructive pulmonary disease. RESULTS: In all, the records of 253 patients with physician-diagnosed chronic obstructive pulmonary disease were analysed. The majority were old (mean age, 78 years). The median number of admissions per patient for this condition in 2010 was two. About two thirds (64%) had had spirometry at least once. Mean forced expiratory volume in one second predicted was 55%. Almost 90% had moderate-to-very severe airflow limitation by spirometry. Overall, long-acting bronchodilators (beta agonists and/or antimuscarinics) were being prescribed for only 21% of the patients. CONCLUSION: Most of the patients admitted to hospital for acute exacerbations of chronic obstructive pulmonary disease were old, had multiple co-morbidities, and the majority had moderate-to-severe airflow limitation by spirometry. Almost half of them (around 46%) had two or more admissions in 2010. Adherence to the latest treatment guidelines seemed inadequate, there being a low prescription rate of long-acting bronchodilators. Chronic obstructive pulmonary disease patients warranting emergency admissions are at risk of future exacerbations and mortality. Management by a designated multidisciplinary team is recommended.

Aspergillus pachymeningitis mimicking nasopharyngeal carcinoma.

OBJECTIVES: We report a case of otogenic fungal pachymeningitis in a diabetic patient who presented with multiple cranial nerve palsies and nasopharyngeal swelling. METHODS: We present a case report, we describe the investigations, management and clinical course of fungal pachymeningitis, and we present a review of the world literature on fungal and non-fungal pachymeningitis. RESULTS: To our knowledge, this is the first report of fungal pachymeningitis with magnetic resonance imaging features suggestive of nasopharyngeal carcinoma. It is also the first reported case with aspergillus cultured from both a dural biopsy and the ear canal. CONCLUSION: Fungal pachymeningitis is a rare condition which may present to otorhinolaryngologists. Its clinical and radiological findings can be confused with those of nasopharyngeal carcinoma; fungal pachymeningitis should thus be included in the differential diagnosis of nasopharyngeal carcinoma.

Reversal of pale-to-dark nasopharyngeal follicle ratio on narrow-band imaging.

Normal nasopharyngeal mucosa contains varying amounts of lymphoid tissue, which in adults may be minimal or absent. Nasopharyngeal mucosa with minimal lymphoid tissue has a regular follicular pattern on narrow-band imaging; pale follicles have thin, dark borders and the ratio of the pale follicle to the dark border (pale-to-dark ratio) is roughly 90%. In some patients undergoing routine nasopharyngeal endoscopy, the pale-to-dark ratio is reversed on narrow-band imaging, with dark centres surrounded by pale borders and a pale-to-dark ratio of roughly 50%. These dark follicles may represent abnormal capillary loops, as they have the same appearance as microvascular changes seen on narrow-band imaging of the oesophageal mucosa which indicate dysplasia or malignancy. While this observed change in the follicular pattern may be an early event in the evolution of nasopharyngeal carcinoma, the significance of this finding remains to be confirmed by a larger-scale study.

Are Hong Kong doctors following the Global Initiative for Asthma guidelines: a questionnaire "Survey on Asthma Management"?

OBJECTIVE: To assess the standard of asthma management by doctors in Hong Kong. DESIGN: Cross-sectional postal questionnaire survey. SETTING: Hong Kong. PARTICIPANTS: Practising doctors registered with the Medical Council of Hong Kong were sent a questionnaire between August and December 2007. MAIN OUTCOME MEASURES: Respondents' responses to questions on demographic data, parameters routinely used to assess asthma control, the pattern of asthma medication prescribing, and seven different case scenarios assessing their ability to classify asthma control and management. RESULTS. We received 410 completed questionnaires from general practitioners (55%), internists (22%), paediatricians (11%), and other specialists (12%). The majority (82%) explained the pathology of asthma to at least some of their patients and tried to identify aggravating factors of the asthma (91%). Fewer observed the inhalation technique of their patients (68%) and prescribed a written asthma management plan (33%). The main medications prescribed to adults and children with asthma were inhaled corticosteroids, inhaled short-acting beta-2 agonists, and combinations of an inhaled corticosteroid and a long-acting beta-2 agonist. In adults and children, long-acting beta-2 agonist alone (without inhaled corticosteroid) was being used to treat asthma by 45% and 36% of the doctors, respectively. Also, 94% of the respondents correctly classified the control status in four out of the seven case scenarios and 31% chose the correct medications when responding to seven of the 14 questions asked. CONCLUSIONS: Asthma management practice of Hong Kong doctors falls short of the standards recommended by international guidelines. More effort in improving their knowledge is urgently warranted.

Regulation of protein expression and function of octn2 in forskolin-induced syncytialization in BeWo Cells.

Placental OCTN2 is a high-affinity carnitine transporter that can interact with a number of therapeutic agents. The process of syncytialization is associated with the expression of a variety of genes. However, the association between syncytialization and OCTN2 expression is not yet clear. Given that forskolin induces BeWo cells to undergo biochemical and morphological differentiation, the purpose of the present study was to investigate whether the function and expression of OCTN2 are influenced by forskolin treatment during syncytialization. The forskolin-induced differentiation of BeWo cells was validated by secretion of beta-human chorionic gonadotropin (beta-hCG) and syncytin expression. Cellular localization of OCTN2 was analyzed by confocal microscopy. Expression of OCTN2 and the modular proteins PDZK1, PDZK2, NHERF1 and NHERF2 was analyzed by Western blotting and carnitine uptake by BeWo cells was estimated and the kinetic properties of uptake measured. The results showed that forskolin treatment increased beta-hCG secretion and syncytin expression, suggesting induction of syncytialization. Confocal images of BeWo cells showed the localization of OCTN2 in the brush-border membrane. OCTN2 protein expression was upregulated in isolated brush-border membranes by long-term forskolin treatment, but the V(m) for carnitine uptake was unchanged, although the K(m) increased. PDZK1, NHERF1 and NHERF2 protein expression in the brush-border membrane was downregulated by forskolin treatment, whereas PDZK2 levels remained unchanged. In conclusion, protein expression and function of OCTN2 in BeWo cells can be regulated by forskolin treatment. While the presence of forskolin results in an increase in OCTN2 protein expression, the increase in uptake capacity may be compensated by the decreased expression of PDZK1, NHERF1 or NHERF2.

MR imaging of nonmalignant polyps and masses of the nasopharynx and sphenoid sinus after radiotherapy for nasopharyngeal carcinoma.

BACKGROUND AND PURPOSE: The development of a new polyp or mass in the radiation field of a previously treated carcinoma is usually an ominous sign of a recurrent cancer, but rarely may it be caused instead by a nonmalignant process. The purpose of this study was to document the MR appearance of unusual nonmalignant polyps or masses (NMPMs) in the nasopharynx and sphenoid sinus arising after radiation treatment of nasopharyngeal carcinoma. MATERIALS AND METHODS: The MR imaging reports of patients undergoing imaging after radiation therapy for nasopharyngeal carcinoma were reviewed retrospectively to identify patients with unusual polyps and masses in the nasopharynx. The MR images of those patients with no evidence of malignancy on biopsy or follow-up were reviewed. RESULTS: The MR imaging reports of 1282 patients were reviewed, and 11 patients (1%) with NMPMs in the nasopharynx or sphenoid sinus were identified. Two patterns were identified: contrast enhancing nasopharyngeal polyps ranging in size from 1 to 5 cm (n = 5) and sphenoid sinus masses consisting of a nonenhancing mass filling a nonexpanded sinus (n = 4) and a heterogeneous enhancing mass expanding the sinus (n = 2). Osteoradionecrosis produced a large defect in the roof of the nasopharynx causing direct communication with the sphenoid sinus (n = 6). Histology revealed granulation tissue in all of the patients with variable amounts of fibrin and inflammatory cells. A direct infective etiology was not proved in any patient. CONCLUSION: NMPMs in the nasopharynx and sphenoid sinus are rare complications after radiation therapy to the skull base, but the radiologist needs to be aware of their appearance so that they can be considered in the differential diagnosis of suspected tumor recurrence.

The frontal bone skull post for image guided surgery of the nasopharynx.

BACKGROUND AND OBJECTIVES: To describe the use of the mastoid skull post of a surgical image guidance system for image guided surgery and intraoperative navigation of the nasopharynx. This adaptation overcomes the limitations of a headset frame or head clamp and can be applied to open and closed surgery of the nasal cavities, sinuses, orbits, midface and anterior skull base. METHODS: A sterile mastoid skull post of a surgical image guidance system was securely screwed into the outer table of the frontal bone and a passive reference frame fixed to it. The position of the reference frame relative to the patient's head remained constant during surgery. RESULTS: Between August 2002 and November 2006, 22 patients underwent a surgical nasopharyngectomy using an image guidance system and frontal bone skull post. No surgical complications or complications of the frontal bone skull post occurred. CONCLUSION: Using the skull post of a surgical image guidance system anchored to the frontal bone to carry a passive reference frame provides real-time visual information about the relationship of relevant anatomy to the surgical field on a fully mobile head in open or closed procedures of the midface and anterior skull base while accuracy and sterility are maintained.

Autoimmune polyendocrinopathy type II in a Chinese patient.

Autoimmune polyendocrinopathy type II is rarely reported in Chinese patients. A 42-year-old Chinese woman with a history of Hashimoto's thyroiditis and hypogonadotropic hypogonadism presented with pneumonia. During hospitalisation, she went into an adrenal crisis and diabetic ketoacidosis. Subsequent dynamic hormonal tests revealed primary and secondary adrenal insufficiency. She also had pernicious anaemia, possible alopecia areata, and myasthenia gravis. This constellation of multiple endocrine and non-endocrine disorders led to the diagnosis of autoimmune polyendocrinopathy type II. As the syndrome can be lethal, it is important to maintain a high index of suspicion, enabling early diagnosis and the appropriate replacement therapy, to ensure a successful outcome.

Focal stereotactic external beam radiotherapy as a vision-sparing method for the treatment of peripapillary and perimacular retinoblastoma: preliminary results.

AIMS: Chemotherapy with aggressive focal ablative therapy is now the mainstay of retinoblastoma therapy. Our experience presents an evolution from conventional radiotherapy by treating posterior pole tumours with focal stereotactic fractionated radiotherapy (SRT). MATERIALS AND METHODS: A retrospective chart review was conducted of five patients (six eyes) treated with SRT at the Hospital for Sick Children and Princess Margaret Hospital, Toronto, Canada, between 1999 and 2004. The prescribed dose was 40 Gy delivered in 20 fractions once daily using 6 MV photons. RESULTS: Five patients (six eyes) were treated. The median age at the time of SRT was 18 months. The median follow-up was 46.5 months as of September 2004. Four patients were treated for a posterior pole focal tumour by focal SRT, and one patient was treated for vitreous seeding with whole-eye SRT. In patients treated with focal SRT, the median doses to the tumour, optic chiasm and brainstem were 41.92, 0.25 and 0.07 Gy, respectively, and to the ipsilateral optic nerve, globe and lens were 9.98, 19.11 and 3.74 Gy, respectively. The median doses to the ipsilateral and contralateral orbital bone were 6.73 Gy (range 5.99-8.29 Gy) and 2.31 Gy (range 0.88-7.08 Gy), respectively. A complete response (residual inactive scar tissue) was seen in four of the five focal tumours treated, with one tumour responding with a partial response (suspicious residual scar tissue). No acute or late side-effects occurred in patients treated with focal SRT. Only the patient treated with whole-eye SRT developed late effects of cataract and corneal ulceration. One patient suffered recurrence within the radiation field 5 months after focal SRT. Control of this recurrence was successful using chemotherapy and focal therapy. No eye has been enucleated. CONCLUSION: Vision-sparing focal SRT for localised tumour masses in critical locations can control tumours with minimal side-effects and a minimal dose to the surrounding critical normal tissue.

Prevalence study of cerebral palsy in Hong Kong children.

OBJECTIVES: To investigate the prevalence of cerebral palsy in local children aged 6 to 12 years and to evaluate service utilisation by those children who attend mainstream schools. DESIGN: Cross-sectional survey. SETTING: Mainstream primary schools and special needs schools in Hong Kong. PARTICIPANTS: Headmasters or headmistresses of special needs schools, and various organisations that provide services to children with cerebral palsy in the school year September 2003 to June 2004. MAIN OUTCOME MEASURES: Prevalence of cerebral palsy and support services used by children with cerebral palsy who attend a mainstream school. RESULTS: Of 435 572 children, 578 with cerebral palsy were identified. The overall point prevalence was 1.3 per 1000 children. The age-specific prevalence rate varied from 1.04 to 1.50 per 1000 children. Approximately 38% of children with cerebral palsy attended a mainstream school. Among those studying in special needs schools, 96% attended a school for the physically handicapped or a school for the severely mentally handicapped. Among 219 children with cerebral palsy in mainstream schools, 57 (26%) received educational support, and 134 (61%) received out-patient therapy support. Only 12% received both supporting services. No educational or therapeutic support was received by 26% of children. CONCLUSIONS: Compared with overseas data, the low prevalence of cerebral palsy detected in local children in this investigation may be due to the differences in study design or a genuinely low prevalence. Setting up a cerebral palsy registry could help monitor the local prevalence of this childhood disability more accurately, thereby providing more reliable information for planning support services for this subgroup of children.

Langmuir-Blodgett-Kuhn and self-assembled films of asymmetrically substituted poly(paraphenylene).

Asymmetrically substituted poly(paraphenylene) (PhPPP) with hydrophilic and hydrophobic side chains was investigated. The polymer behavior at the air-water interface was studied on the basis of surface pressure-area (pi-A) isotherms and compression/expansion hysteresis measurements. PhPPP can form stable monolayers with an area per repeat unit of A=0.20+/-0.02 nm2 and a collapse pressure in the range of pi=25 mN/m. Then, Langmuir-Blodgett-Kuhn (LBK) films of PhPPP were prepared by horizontally and vertically transferring the Langmuir monolayers onto hydrophilic solid substrates at pi=12 mN/m. Cross-section analysis of the AFM tapping-mode topography images of a single transferred monolayer reveals a thickness of d0=0.9+/-0.1 nm. Taking into account the obtained monolayer thickness, curve-fitting calculations of angular scan data of LB monolayers measured using surface plasmon resonance (SPR) spectroscopy lead to a value for the refractive index of n=1.78+/-0.02 at lambda=632.8 nm. Next, the spontaneous formation of a PhPPP monolayer by adsorption from solution was studied ex situ by atomic force microscopy and UV-vis spectroscopy and in situ by using SPR spectroscopy. Stable self-assembled monolayers of PhPPP can be formed on hydrophilic surfaces with a thickness similar to that of the monolayer obtained using the LB method. The characterization results confirmed the amphiphilic character and the self-assembly properties of PhPPP, as well as the possibility of preparing homogeneous monolayer and multilayer films.

Neuroimpairment, activity limitation, and participation restriction among children with cerebral palsy in Hong Kong.

OBJECTIVES: To study children with cerebral palsy in Hong Kong, their neuroimpairment, activity limitation, and participation restriction in society. Parents' opinion on current medical and rehabilitation services was also sought. DESIGN: Systematic survey using questionnaires. SETTING: Four associations in Hong Kong: Child Assessment Service, Hong Kong Association for Parents of Children with Physical Disabilities, Association of Parents of the Severely Mentally Handicapped, and Hong Kong Physically Handicapped and Able-Bodied Association. PARTICIPANTS: Parents of children with cerebral palsy. MAIN OUTCOME MEASURES: Neuroimpairment, activity limitation, and participation restriction. RESULTS: Information from 181 children with cerebral palsy was analysed. Among them, 56% were boys. The mean age was 7 years 6 months (standard deviation, 3 years 11 months). The most common diagnostic type was spastic cerebral palsy. Co-morbidities in children with cerebral palsy were common. Limitation in daily activities including mobility and self-care tasks was considerable and this posed great stress to parents when taking care of their children. Children's participation in both social and leisure activities was regarded as a low priority. A high percentage (70%) of parents reported difficulty in travelling. The reasons involved problems in transportation, building access (entry and exit), and attitudes of the general public. These environmental factors restricted the social participation of the children and their families. Over 75% of parents were satisfied with the current medical and rehabilitation services. CONCLUSIONS: Children with cerebral palsy have multiple and complex needs. The findings of this study may serve as a reference for parents, service providers, and policy makers to work in partnership to achieve a more comprehensive health-care service for children with cerebral palsy and to facilitate better integration into the community.

Concurrent chemoradiotherapy followed by surgery in locally advanced squamous cell carcinoma of the oesophagus: a single centre experience.

INTRODUCTION: Data on combined modality treatment for locally advanced squamous cell carcinoma of the oesophagus involving Asian patients are limited. MATERIALS AND METHODS: A retrospective study of 56 consecutive patients with this condition treated with concurrent chemoradiotherapy followed by surgery in a single tertiary institution in Singapore was performed. RESULTS: The median overall survival of the entire cohort was 14.1 months [95% confidence interval (CI); range, 8.6 to 19.6 months]. In patients who underwent successful oesophagectomy after chemoradiotherapy (n = 17), the median survival was 27.8 months compared to 9.8 months for those who did not have surgery (n = 39) (P = 0.046, log-rank test). The median time to first relapse for the entire cohort was 16.1 months (95% CI, 7.7 to 24.5 months). The time to first relapse was 23.9 months in the subgroup of patients with successful surgery and 12.1 months in the group which did not (P = 0.147, log-rank test). The high proportion of patients who were medically unfit for surgery or declined surgery may have conferred a selection bias. CONCLUSION: Concurrent chemoradiotherapy followed by surgery is feasible in selected patients. The benefit of adding of surgery to chemoradiotherapy is still controversial and we await the results of randomised controlled trials comparing chemoradiotherapy with surgery versus chemoradiotherapy alone.

Prevalence of Helicobacter pylori in gastric cancer in a South-East Asian population by 14C-urea breath test.

BACKGROUND: Helicobacter pylori is believed to play an important role in the aetiology of gastric cancer. There is a great variability in seropositivity and histological frequency of H. pylori in gastric cancer. The present prospective study investigates the prevalence of H. pylori infection in gastric cancer patients using 14C-urea breath testing. METHODS: Patients with endoscopic biopsy-proven gastric cancer were fasted for 6 h prior to ingesting 18.5 x 104 Bq of 14C-urea cocktail orally. Breath samples were collected after 20 min by asking them to blow into a hyamine solution and measurements were read in a scintillation counter. RESULTS: Fifty out of 51 patients (98%) with gastric cancer were positive on the 14C-urea breath test compared to 29 patients (61%) who were positive on histology. There was no association between sex, age or tumour site, stage, differentiation, Lauren type and H. pylori status. The test was negative in one patient with cardial tumour in which histology of the resected specimen was also negative for the bacteria. CONCLUSIONS: Active H. pylori infection is highly prevalent in gastric cancer in a South-East Asian population. The 14C-urea breath test is a highly sensitive method for detecting the presence of H. pylori even in gastric adenocarcinoma irrespective of the stage.

Conformational propagation with prion-like characteristics in a simple model of protein folding.

Protein refolding/misfolding to an alternative form plays an aetiologic role in many diseases in humans, including Alzheimer's disease, the systemic amyloidoses, and the prion diseases. Here we have discovered that such refolding can occur readily for a simple lattice model of proteins in a propagatable manner without designing for any particular alternative native state. The model uses a simple contact energy function for interactions between residues and does not consider the peculiarities of polypeptide geometry. In this model, under conditions where the normal (N) native state is marginally stable or unstable, two chains refold from the N native state to an alternative multimeric energetic minimum comprising a single refolded conformation that can then propagate itself to other protein chains. The only requirement for efficient propagation is that a two-faced mode of packing must be in the ground state as a dimer (a higher-energy state for this packing leads to less efficient propagation). For random sequences, these ground-state dimeric configurations tend to have more beta-sheet-like extended structure than almost any other sort of dimeric ground-state assembly. This implies that propagating states (such as for prions) are beta-sheet rich because the only likely propagating forms are beta-sheet rich. We examine the details of our simulations to see to what extent the observed properties of prion propagation can be predicted by a simple protein folding model. The formation of the alternative state in the present model shows several distinct features of amyloidogenesis and of prion propagation. For example, an analog of the phenomenon of conformationally distinct strains in prions is observed. We find a parallel between 'glassy' behavior in liquids and the formation of a propagatable state in proteins. This is the first report of simulation of conformational propagation using any heteropolymer model. The results imply that some (but not most) small protein sequences must maintain a sequence signal that resists refolding to propagatable alternative native states and that the ability to form such states is not limited to polypeptides (or reliant on regular hydrogen bonding per se) but can occur for other protein-like heteropolymers.

Typical and atypical Carney's triad presenting with malignant hypertension and papilledema.

This report encourages clinicians to consider a diagnosis of Carney's triad in patients with multifocal gastric stromal sarcoma, extraadrenal paraganglioma (predominantly mediastinal), or pulmonary chondroma. The authors conducted a retrospective 20-year survey at the Hospital for Sick Children and identified two children with Carney's triad. One child, presenting atypically with papilledema and fundal hemorrhages from malignant hypertension and benign intracranial hypertension from chronic iron-deficiency anemia, is the second patient ever to date be described with the complete Carney's triad of neoplasms at diagnosis. Another child presented more typically with gastric stromal sarcoma and pulmonary chondroma without paraganglioma. Carney's triad is a rare differential diagnosis for "idiopathic" hypertension or iron-deficiency anemia from chronic gastrointestinal bleeding. If missed, patients with Carney's triad may have the debilitating physical and mental consequences of chronic iron deficiency and may die of untreated prolonged hypertension and metastatic leiomyosarcoma.