Anatomical tilt lateral wrist X-ray - Not always 22°.

INTRODUCTION: Open Reduction and Internal Fixation (ORIF) with volar locking plates are commonly used to manage distal radial fractures. The anatomical tilt lateral (ATL) wrist X-ray is often required for evaluation of intra-articular screw penetration due to the screw position. This study aims to evaluate the correlation between the tube angulation given by performing radiographers for the ATL projection and the post-examination measurement of radial inclination (RI) on the Posterior Anterior (PA) wrist image. METHODS: A retrospective review was performed for 36 patients. A standardised method developed by Kreder et al. (1996) was used to measure the RI on the PA wrist image. All ATL images sent into Picture Archiving and Communications System (PACS) have the tube angulation applied annotated on the image. Pearson's correlation was used to analyse the co-relationship between the RI and the tube angle applied for ATL projection. RESULTS: The average angle of RI measured by the four observers was 19. Normality of 0.385 was established. A positive correlation (p = 0.792) between the RI and the tube angle applied for ATL was found. CONCLUSION: Our study found a strong positive correlation between the tube angulation applied by performing radiographers for the ATL projection and the post-examination RI measured on the PA wrist image by the independent reviewers. This suggests that radiographers can use the measured RI to apply the tube angulation when performing the ATL wrist X-ray, instead of estimating the tube angulation to be applied. IMPLICATIONS FOR PRACTICE: Using the measured RI to apply the tube angulation when performing the ATL wrist X-ray will ensure a more reliable and reproducible way that could reduce the number of repeated images and, thus, unnecessary radiation dose to patients.

Latent class analysis of structured histopathology in prognosticating surgical outcomes of chronic rhinosinusitis with nasal polyps in Singapore.

BACKGROUND: Structured histopathology profiling is recommended when reporting chronic rhinosinusitis with nasal polyp (CRSwNP) tissue. The objective of this study is to identify features in structured histopathology that predict outcome after functional endoscopic sinus surgery (FESS) in a cohort of CRSwNP patients from Singapore. METHODS: Latent class analysis was performed on structured histopathology reports of 126 CRSwNP patients who had undergone FESS. Outcome measures were polyp recurrence, need for systemic corticosteroids, revision surgery or biologics, and disease control at 2 years post-FESS. RESULTS: Three classes were identified. Class 1 was characterised by mild, predominantly lymphoplasmacytic inflammation. Class 2 comprised of 100 eosinophils/HPF, hyperplastic seromucinous glands, mucosal ulceration and mucin containing eosinophil aggregates and Charcot-Leyden crystals. Classes 2 and 3 were significantly associated with uncontrolled disease at 2 years post-FESS. Class 3 was additionally associated with the need for systemic corticosteroids. CONCLUSIONS: Eosinophil count, degree of inflammation, predominant inflammatory type, hyperplastic seromucinous glands, mucosal ulceration and mucin containing eosinophil aggregates and Charcot-Leyden crystals predicted need for systemic corticosteroids and uncontrolled disease at 2 years post-FESS. The presence of >100 eosinophils/HPF should be reported, as this subset of tissue eosinophilia was associated with less favourable outcomes after FESS.

Nutritional Status and Health-Related Quality of Life among Knee and Hip Osteoarthritis Patients under Rehabilitation Care in Kuala Nerus, Terengganu, Malaysia.

INTRODUCTION: The World Health Organisation (WHO) has estimated that 80% of people with osteoarthritis (OA) have movement limitations while 25% of them cannot perform their major daily activities, thus resulting in a decline of their nutritional status and quality of life (QOL). Therefore, this study aimed to compare the nutritional status and health-related quality of life (HRQOL) of OA patients between gender and age group. MATERIAL AND METHODS: A cross-sectional study was conducted on 131 OA patients in Rehabilitation Health Organisation, Terengganu. Socio-demographic, clinical, lifestyle histories, 24-hour dietary intake and HRQOL were assessed using a structured questionnaire. RESULTS: Knee and/or hip OA patients recruited consisted of 19.1% of men and 80.9 % of women collectively with a mean age of 61.81 (9.28) years ranging from 38 to 83 years. The percentages of underweight, normal, overweight, and obese patients were 1.5%, 12.2%, 36.7%, and 49.6%, respectively. Further assessment of HRQOL showed that the highest mean score was obtained by the social functioning (SF) domain of 41.25 (27.16), while the mental domain scored the least mean score of 21.15 (20.92). In terms of gender breakdown, the males had significantly greater weight and height but lower body fat (BF) compared to their female counterparts, as well as a significantly higher energy, carbohydrate and protein intake. According to the age group, patients aged < 60 years had significantly greater weight, height, and BF than those aged ≥ 60 years. CONCLUSION: This study is an important baseline reference for proper OA management and prevention by providing crucial nutritional status and HRQOL information.

Factors affecting surgical mortality of oral squamous cell carcinoma resection.

Survival rates for oral squamous cell carcinoma (OSCC) has remained stagnant in recent years and improving surgical mortality could be an avenue to enhance outcomes. This systematic review aims to identify the causes of mortalities, determine both the modifiable and non-modifiable factors involved and target a reduction in postoperative 30-day mortality. In May 2019, a comprehensive search of key databases including PubMed, EMBASE, Cochrane Library was conducted. Blinded selection by two researchers identified papers that included participants who received oral squamous cell carcinoma resection and suffered an in-hospital or 30-day mortality. Selection identified two relevant papers that meet the inclusion criteria. One study had one death in its population sample but only had the cause of death described. Another study had an overall surgical mortality rate of 1% in a population of 21,681. Patients with multiple factors had the highest mortality rates; 4.6% in patients >85 years old and have a T4 diagnosis, 3.9% in patients with a Comorbidity Index ≥1 and a T4 diagnosis. These studies did not determine relationships between factors and causes of death. There are significant knowledge gaps in the literature, that can be addressed through further population analysis studies.

Predictors of acute neurological complication following tetralogy of Fallot operation in Serdang Hospital, Malaysia.

BACKGROUND: The long waiting time for Tetralogy of Fallot (TOF) operation may potentially increase the risk of hypoxic insult. Therefore, the objective of this study is to determine the frequency of acute neurological complications following primary TOF repair and to identify the peri-operative risk factors and predictors for the neurological sequelae. METHODS: A retrospective review of the medical and surgical notes of 68 patients who underwent TOF repair in Hospital Serdang, from January 2013 to December 2017 was done. Univariate and multivariate analyses of demographics and perioperative clinical data were performed to determine the risk for the development of acute neurological complications (ANC) among these patients. RESULTS: ANC was reported in 13 cases (19.1%) with delirium being the most common manifestation (10/68, 14.7%), followed by seizures in 4 (5.9%) and abnormal movements in two patients (2.9%). Univariate analyses showed that the presence of right ventricular (RV) dysfunction, prolonged duration of inotropic support (≥7 days), prolonged duration of mechanical ventilation (≥7 days), longer length of ICU stays (≥7 days), and longer length of hospital stay (≥14 days), were significantly associated with the presence of ANCs (p<0.05). However, multivariate analyses did not show any significant association between these variables and the development of ANC (p>0.05). The predictors for the development of postoperative delirium were pre-operative oxygen saturation less than 75% (Odds Ratio, OR=16.90, 95% Confidence Interval, 95%CI:1.36, 209.71) and duration of ventilation of more than 7 days (OR=13.20, 95%CI: 1.20, 144.98). CONCLUSION: ANC following TOF repair were significantly higher in patients with RV dysfunction, in those who required a longer duration of inotropic support, mechanical ventilation, ICU and hospital stay. Low pre-operative oxygen saturation and prolonged mechanical ventilation requirement were predictors for delirium which was the commonest neurological complications observed in this study. Hence, routine screening for delirium using an objective assessment tool should be performed on these high-risk patients to enable accurate diagnosis and early intervention to improve the overall outcome of TOF surgery in this country.

Using a checklist to assess if a child undergoing MRI needs general anaesthesia.

AIM: To assess if a child-assessment checklist covering tasks children are expected to perform during magnetic resonance imaging (MRI) can determine whether the child requires general anaesthesia (GA) during MRI. MATERIALS AND METHODS: In this institute review board approved study, children who underwent MRI from September 2016 to June 2017 at KK Women's and Children's Hospital were assessed using a checklist by a research assistant before their examination. During this project, the checklist had no influence on whether the MRI was performed under GA or not. The checklist consisted of five items rated on a binary scale assessing the child's behaviour. Binary logistic regression was performed separately on the overall sample and for a subset of younger children to identify variables associated with the requirement for GA. RESULTS: The mean age of the overall sample (798 children) and the subset of children <8 years (124 children) were 11.7±3.7 and 5.5±1.3 years, respectively. In both groups, children who required GA were significantly younger than those who did not (p<0.001). No gender differences were observed. Children who required GA scored higher on the checklist compared to those who did not in both groups (p<0.001). The diagnostic accuracy of the checklist was found to be good (area under the curve [AUC]=0.97 for both groups), with a suggested cut-off score of 4. Intraclass correlation coefficient of the ratings by two independent individuals was 0.78. CONCLUSION: The child assessment checklist was useful in identifying GA requirement in children undergoing MRI and can be administered by non-medical staff with good inter-rater reliability.

Deubiquitinase USP10 regulates Notch signaling in the endothelium.

Notch signaling is a core patterning module for vascular morphogenesis that codetermines the sprouting behavior of endothelial cells (ECs). Tight quantitative and temporal control of Notch activity is essential for vascular development, yet the details of Notch regulation in ECs are incompletely understood. We found that ubiquitin-specific peptidase 10 (USP10) interacted with the NOTCH1 intracellular domain (NICD1) to slow the ubiquitin-dependent turnover of this short-lived form of the activated NOTCH1 receptor. Accordingly, inactivation of USP10 reduced NICD1 abundance and stability and diminished Notch-induced target gene expression in ECs. In mice, the loss of endothelial Usp10 increased vessel sprouting and partially restored the patterning defects caused by ectopic expression of NICD1. Thus, USP10 functions as an NICD1 deubiquitinase that fine-tunes endothelial Notch responses during angiogenic sprouting.

Adverse outcomes after planned surgery with anticipated intensive care admission in out-of-office-hours time periods: a multicentre cohort study.

BACKGROUND: Increasing mortality for patients admitted to hospitals during the weekend is a contentious but well described phenomenon. However, it remains uncertain whether adverse outcomes, including prolonged hospital length-of-stay (LOS), may also occur after patients undergoing major planned surgery are admitted to an intensive care unit (ICU) out-of-office-hours, either during weeknights (after 18:00) or on weekends. METHODS: All planned surgical admissions requiring admission to one of 183 ICUs across Australia and New Zealand between 2006 and 2016 were included in this retrospective population-based cohort study. Primary outcomes were hospital LOS and hospital mortality. RESULTS: Of the total 504 713 planned postoperative ICU admissions, 33.6% occurred during out-of-office-hours. After adjusting for available risk factors, out-of-office-hours ICU admissions were associated with a significant increase in hospital LOS [+2.6 days, 95% confidence interval (CI) 2.5-2.6], mortality [odd ratio (OR) 1.5, 95%CI 1.4-1.6], and a reduced chance of being directly discharged home (OR 0.8, 95%CI 0.8-0.8). The strongest association for adverse outcomes occurred with weekend ICU admissions (hospital LOS: +3.0 days, 95%CI 3.2-3.6; hospital mortality: OR 1.7, 95%CI 1.6-1.8). Clustering of adverse outcomes by hospitals was not observed in the generalised estimating equation analyses. CONCLUSIONS: Despite a greater clinical staff availability and higher monitoring levels, planned surgery requiring anticipated out-of-office-hours ICU admission was associated with a prolonged hospital LOS, reduced discharge directly home, and increased mortality compared with in-office-hours admissions. Our findings have potential clinical, economic and health policy implications on how complex planned surgery should be planned and managed.

Role of IL-13Rα2 in modulating IL-13-induced MUC5AC and ciliary changes in healthy and CRSwNP mucosa.

BACKGROUND: The IL-13 receptor α2 (IL-13Rα2) is a receptor for IL-13 which has conflicting roles in mediating IL-13 responses in the lower airway, with little known about its impact on upper airway diseases. We sought to investigate the expression of IL-13 receptors, IL-13Rα1 and IL-13Rα2, in chronically inflamed nasal epithelium, and explore IL-13-induced signaling pathways in an in vitro model of human nasal epithelial cells (hNECs). METHODS: The protein and mRNA expression levels of IL-13 and its receptors in nasal biopsies of patients with nasal polyps (NP) and healthy controls were evaluated. We investigated goblet cell stimulation with mucus hypersecretion induced by IL-13 (10 ng/mL, 72 hours) treatment in hNECs using a pseudostratified epithelium in air-liquid interface (ALI) culture. RESULTS: There were significant increases in IL-13, IL-13Rα1, and IL-13Rα2 mRNA and protein levels in NP epithelium with healthy controls as baseline. MUC5AC mRNA positively correlated with IL-13Rα2 (r = .5886, P = .002) but not with IL-13Rα1 in primary hNECs. IL-13 treatment resulted in a significant increase in mRNA and protein levels of IL-13Rα2 only in hNECs. IL-13 treatment induced an activation of extracellular signal-regulated kinases (ERK)1/2 and an upregulation of C-JUN, where the IL-13-induced effects on hNECs could be attenuated by ERK1/2 inhibitor (50 µmol/L) or dexamethasone (10-4 -10-7  mol/L) treatment. CONCLUSIONS: IL-13Rα2 has a potential role in IL-13-induced MUC5AC and ciliary changes through ERK1/2 signal pathway in the nasal epithelium. IL-13Rα2 may contribute to airway inflammation and aberrant remodeling which are the main pathological features of CRSwNP.

Glutamic acid decarboxylase and islet antigen 2 antibody profiles in people with adult-onset diabetes mellitus: a comparison between mixed ethnic populations in Singapore and Germany.

AIM: To gain insight into the presence of islet cell autoimmunity in an ethnic Asian compared with a white European population. METHODS: For this cross-sectional study we recruited people with adult-onset diabetes (age of diagnosis 20-60 years), at tertiary referral centres in Germany (n=1020) and Singapore (n=1088). Glutamic acid decarboxylase and islet antigen 2 antibodies were measured according to Islet Autoantibody Standardization Program protocols. RESULTS: The prevalence of glutamic acid decarboxylase antibody positivity was 13.9% (95% CI 12.1-16.0; P<0.001) in the white European cohort compared with 6.8% (95% CI 5.5-8.4; P<0.001) in the Asian cohort. Glutamic acid decarboxylase antibody positivity was 11.4% (95% CI 7.7-16.6) in Indian, 6.0% (95% CI 3.6-9.9) in Malay and 5.8% (95% CI 4.3-7.7; P<0.001) in Chinese participants. In the white European participants, the prevalence of islet antigen 2 antibody positivity was 7.8% (95% CI 6.4-9.4) compared with 14.8% (95% CI 12.8-17.0; P<0.001) in the Asian cohort as a whole, and among the three ethnicities in the Asian cohort it was 12.4% (95% CI 8.6-17.7) in Indian, 16.8% (95% CI 12.6-22.2) in Malay and 15.7% (95% CI 13.2-18.6) in Chinese participants. Double antibody positivity was seen in 5.7% (95% CI 4.5-7.1) of white European participants compared with 1.6% (95% CI 1.0-2.5; P<0.01) of Asian participants. In the white European cohort, those who were glutamic acid decarboxylase autoantibody-positive had a lower BMI than those who were autoantibody-negative, but this trend was absent in the Asian cohort. CONCLUSIONS: A marked prevalence of islet cell autoimmunity was observed in people with adult-onset diabetes. While glutamic acid decarboxylase antibodies were more frequent in the European cohort, islet antigen 2 antibody positivity was highest in the three ethnic groups in Singapore, suggesting ethnic-specific differences in antibody profiles.

Profile differences between overdose and non-overdose suicide attempts in a multi-ethnic Asian society.

BACKGROUND: This study explores differences in characteristics of overdose (OD) and non-overdose (NOD) suicide attempts in Singapore. METHODS: Four hundred eighty-five medical records of people who attempted suicide were extracted from a local general hospital patient database and classified into OD and NOD groups. Differences in socio-demographic factors, suicide characteristics and hospital admission types between both groups were examined. RESULTS: Indians were more likely than the Chinese and Malays to employ OD method in their attempts. More suicide attempts in the OD group than NOD group were self-reported. The most likely place for suicide attempts for both groups was at home, though more NOD suicide attempts were in public areas as compared to the OD group. Analgesics were the most used substance in the OD group. Those who attempted suicide using OD had a higher number of psychiatric ward admissions than the NOD group. Risk and protective factors varied between both groups. CONCLUSION: Differences in socio-demographics, suicide characteristics and admission characteristics between OD and NOD groups were observed. Recommendations for suicide prevention in the community are discussed. Further studies on the mediators and moderators of these trends and characteristics of suicide attempts are necessary to ensure maximal efficacy of prevention and management.

The mouse endometrium contains epithelial, endothelial and leucocyte populations expressing the stem cell marker telomerase reverse transcriptase.

STUDY HYPOTHESIS: The mouse endometrium harbours stem/progenitor cells that express the stem cell marker mouse telomerase reverse transcriptase (mTert). STUDY FINDING: We used a mouse carrying a transgenic reporter for mTert promoter activity to identify rare endometrial populations of epithelial and endothelial cells that express mTert. WHAT IS KNOWN ALREADY: Stem/progenitor cells are hypothesized to be responsible for the remarkable regenerative capacity of the endometrium, but the lack of convenient endometrial stem/progenitor markers in the mouse has hampered investigations into the identity of these cells. STUDY DESIGN, SAMPLES/MATERIALS, METHODS: A mouse containing a green fluorescent protein (GFP) reporter under the control of the telomerase reverse transcriptase promoter (mTert-GFP) was used to identify potential stem/progenitor cells in the endometrium. mTert promoter activity was determined using fluorescence microscopy and flow cytometry to identify GFP(+) cells. GFP(+) cells were examined for epithelial, stromal, endothelial, leucocyte and proliferation markers and bromodeoxyuridine retention to determine their identity. The endometrium of ovariectomized mice was compared to that of intact cycling mice to establish the role of ovarian hormones in maintaining mTert-expressing cells. MAIN RESULTS AND THE ROLE OF CHANCE: We found that mTert-GFP is expressed by rare luminal and glandular epithelial cells (0.3% of epithelial cells by flow cytometry), rare CD45(-) cells in the stromal compartment (0.028 ± 0.010% of stromal cells by microscopy) and many CD45(+) leucocytes. Ovariectomy resulted in significant decrease of mTert-GFP(+) epithelial cells (P = 0.029 for luminal epithelium; P = 0.034 for glandular epithelium) and a decrease in the percentage of mTert-GFP(+) CD45(+) leucocytes in the stromal compartment (P = 0.015). However, CD45(-) mTert-GFP(+) cells in the stromal compartment were maintained in ovariectomized mice. This population is enriched for cells bearing the endothelial marker CD31 (10.3% of CD90(-) CD45(-) and 97.8% CD90(+) CD45(-) by flow cytometry). CD45(-) mTert-GFP(+) cells also immunostained for the endothelial marker von Willebrand factor. These results suggest that the endometrial epithelium and vasculature are foci of stem/progenitor activity and provide a system to investigate molecular mechanisms involved in endometrial regeneration and repair. LIMITATIONS, REASONS FOR CAUTION: The stem/progenitor activity of endometrial mTert-GFP(+) cells needs to be experimentally verified. WIDER IMPLICATIONS OF THE FINDINGS: The identification and characterization of mTert-expressing progenitor cells in the mouse will facilitate the identification of equivalent populations in the human endometrium that are likely to be involved in endometrial function, fertility and disease. LARGE-SCALE DATA: Not applicable. STUDY FUNDING AND COMPETING INTERESTS: This study was funded by National Health and Medical Research Council (NHMRC) of Australia grants (1085435, C.E.G., J.A.D.), 1021127 (C.E.G.), NHMRC Senior Research Fellowship (1042298, C.E.G.), the Victorian Infrastructure Support Program, U.S. National Institutes of Health grant R01 DK084056 (D.T.B.) and the Harvard Stem Cell Institute (D.T.B.). The authors have no conflicts of interest to declare.

Knowledge, attitude and practice on standard precautions for prevention of HIV infection among clinical year medical students.

BACKGROUND: Human Immunodeficiency Virus (HIV) can be transmitted through blood, vaginal secretion, infected semen, breast milk as well as blood containing saliva, vomitus and urine. Health care workers (HCWs) are at risk of HIV infection; and standard precautions is a guideline to be followed by HCWs to prevent it. OBJECTIVE: This study was aimed to evaluate the level of knowledge, attitude and practice on standard precautions for prevention of HIV infection; and its associated factors. MATERIALS AND METHODS: This cross-sectional study was conducted among 200 clinical year medical students from a public university in Malaysia. The clinical year medical students were arranged into strata according to year of study and were randomly selected via stratified random sampling. Each respondent were provided a selfadministered questionnaire. There were four sections in the questionnaire to obtain information on socio-demographic characteristics, knowledge, attitude and practice on standard precautions. Both descriptive and analytical analyses such as Chi-squared test were performed. RESULTS: A total of 162 respondents participated in this study, contributed to the response rate of 81%. The study demonstrated that there was no significant association between level of practice with socio-demographic characteristics such as gender, ethnicity, age, religion, year of study and total family income. However, there was a significant association between level of practice with level of knowledge and attitude (p<0.05). For every one year increase in age, the respondents were 1.7 times (p=0.001) and 1.5 times (p=0.012) more likely to have knowledge score between 50th and 75th percentile and above 75th percentile compared to below 50th percentile, respectively. CONCLUSION: It could be interpreted from the findings, that there is a need for further improvement in the knowledge and attitude level among the respondents; which will eventually improve their practice.

Sequence analysis of E/NS1 gene junction of Dengue Type-1 viruses isolated in Klang Valley 2010 to 2012.

Dengue is a mosquito-borne viral disease caused by four serotypes of dengue virus, affecting the human population for decades in many tropical and subtropical regions of the world. In Malaysia, all four dengue serotypes co-circulates in a dengue season even though any one of the serotypes can predominate. In this study, serum samples were collected from dengue fever and severe dengue fever patients within Klang Valley from 2010-2012 to determine the prevailing dengue serotypes. In addition, sequencing of the envelope/nonstructural 1 (E/NS1) gene junction of the virus isolated was performed to identify the presence of any mutations that are suggestive of increased virulence in the virus. The results showed that Dengue-1 (DEN-1) was the predominant circulating serotype. The E/NS1 gene sequences of the isolates were analysed to trace the evolutionary knowledge of the strains. All sequences of the isolates were compared with DEN-1 prototype Hawaii strain as the reference sequence. The E/NS1 sequences of other dengue strains from neighbouring regions as well as other parts of the world obtained from the GenBank database were also included in the phylogenetic tree analysis. Analyses showed that there was 97% to 100% similarity among the ten isolates at the nucleotide level. Similarly, the amino acid analogue also showed 98% to 100% homology. However, all five non-severe dengue isolates showed variation at position 780, resulting in an amino acid change from valine to alanine as compared to severe dengue isolates. A rooted phylogenetic tree was performed using neighbour-joining method with DEN-2 and DEN-3 as the outgroups. Results showed that all ten isolates were classified as genotype I. In addition, the five isolates from severe dengue patients were found to be clustered together with JN697057 and JN697058, Malaysian DEN-1 strains from the 2005 outbreak.

Isolation and characterisation of mesenchymal stem/stromal cells in the ovine endometrium.

OBJECTIVE: Mesenchymal stem/stromal cells (MSC) were recently discovered in the human endometrium. These cells possess key stem cell properties and show promising results in small animal models when used for preclinical tissue engineering studies. A small number of surface markers have been identified that enrich for MSC from bone marrow and human endometrium, including the Sushi Domain-containing 2 (SUSD2; W5C5) and CD271 markers. In preparation for developing a large animal preclinical model for urological and gynecological tissue engineering applications we aimed to identify and characterise MSC in ovine endometrium and determine surface markers to enable their prospective isolation. MATERIALS AND METHODS: Ovine endometrium was obtained from hysterectomised ewes following progesterone synchronisation, dissociated into single cell suspensions and tested for MSC surface markers and key stem cell properties. Purified stromal cells were obtained by flow cytometry sorting with CD49f and CD45 to remove epithelial cells and leukocytes respectively, and MSC properties investigated. RESULTS: There was a small population CD271+ stromal cells (4.5 ± 2.3%) in the ovine endometrium. Double labelling with CD271 and CD49f showed that the sorted CD271+CD49f- stromal cell population possessed significantly higher cloning efficiency, serial cloning capacity and a qualitative increased ability to differentiate into 4 mesodermal lineages (adipocytic, smooth muscle, chondrocytic and osteoblastic) than CD271-CD49f- cells. Immunolabelling studies identified an adventitial perivascular location for ovine endometrial CD271+ cells. CONCLUSION: This is the first study to characterise MSC in the ovine endometrium and identify a surface marker profile identifying their location and enabling their prospective isolation. This knowledge will allow future preclinical studies with a large animal model that is well established for pelvic organ prolapse research.

Restriction fragment mass polymorphism (RFMP) analysis based on MALDI-TOF mass spectrometry for detecting antiretroviral resistance in HIV-1 infected patients.

Viral genotype assessment is important for effective clinical management of HIV-1 infected patients, especially when access and/or adherence to antiretroviral treatment is reduced. In this study, we describe development of a matrix-assisted laser desorption/ionization-time of flight mass spectrometry-based viral genotyping assay, termed restriction fragment mass polymorphism (RFMP). This assay is suitable for sensitive, specific and high-throughput detection of multiple drug-resistant HIV-1 variants. One hundred serum samples from 60 HIV-1-infected patients previously exposed to nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs) and protease inhibitors (PIs) were analysed for the presence of drug-resistant viruses using the RFMP and direct sequencing assays. Probit analysis predicted a detection limit of 223.02 copies/mL for the RFMP assay and 1268.11 copies/mL for the direct sequencing assays using HIV-1 RNA Positive Quality Control Series. The concordance rates between the RFMP and direct sequencing assays for the examined codons were 97% (K65R), 97% (T69Ins/D), 97% (L74VI), 97% (K103N), 96% (V106AM), 97% (Q151M), 97% (Y181C), 97% (M184VI) and 94% (T215YF) in the reverse transcriptase coding region, and 100% (D30N), 100% (M46I), 100% (G48V), 100% (I50V), 100% (I54LS), 99% (V82A), 99% (I84V) and 100% (L90M) in the protease coding region. Defined mixtures were consistently and accurately identified by RFMP at 5% relative concentration of mutant to wild-type virus while at 20% or greater by direct sequencing. The RFMP assay based on mass spectrometry proved to be sensitive, accurate and reliable for monitoring the emergence and early detection of HIV-1 genotypic variants that lead to drug resistance.

Long-term performance evaluation of EBPR process in tropical climate: start-up, process stability, and the effect of operational pH and influent C:P ratio.

To date, little information is known about the operation of the enhanced biological phosphorus removal (EBPR) process in tropical climates. Along with the global concerns on nutrient pollution and the increasing array of local regulatory requirements, the applicability and compliance accountability of the EBPR process for sewage treatment in tropical climates is being evaluated. A sequencing batch reactor (SBR) inoculated with seed sludge from a conventional activated sludge (CAS) process was successfully acclimatized to EBPR conditions at 28 °C after 13 days' operation. Enrichment of Candidatus Accumulibacter phosphatis in the SBR was confirmed through fluorescence in situ hybridization (FISH). The effects of operational pH and influent C:P ratio on EBPR were then investigated. At pH 7 or pH 8, phosphorus removal rates of the EBPR processes were relatively higher when operated at C:P ratio of 3 than C:P ratio of 10, with 0.019-0.020 and 0.011-0.012 g-P/g-MLVSS•day respectively. One-year operation of the 28 °C EBPR process at C:P ratio of 3 and pH 8 demonstrated stable phosphorus removal rate of 0.020 ± 0.003 g-P/g-MLVSS•day, corresponding to effluent with phosphorus concentration <0.5 mg/L. This study provides the first evidence on good EBPR activity at relatively high temperature, indicating its applicability in a tropical climate.