Progesterone receptor mediates ovulatory transcription through RUNX transcription factor interactions and chromatin remodelling.

Progesterone receptor (PGR) plays diverse roles in reproductive tissues and thus coordinates mammalian fertility. In the ovary, rapid acute induction of PGR is the key determinant of ovulation through transcriptional control of a unique set of genes that culminates in follicle rupture. However, the molecular mechanisms for this specialized PGR function in ovulation is poorly understood. We have assembled a detailed genomic profile of PGR action through combined ATAC-seq, RNA-seq and ChIP-seq analysis in wildtype and isoform-specific PGR null mice. We demonstrate that stimulating ovulation rapidly reprograms chromatin accessibility in two-thirds of sites, correlating with altered gene expression. An ovary-specific PGR action involving interaction with RUNX transcription factors was observed with 70% of PGR-bound regions also bound by RUNX1. These transcriptional complexes direct PGR binding to proximal promoter regions. Additionally, direct PGR binding to the canonical NR3C motif enable chromatin accessibility. Together these PGR actions mediate induction of essential ovulatory genes. Our findings highlight a novel PGR transcriptional mechanism specific to ovulation, providing new targets for infertility treatments or new contraceptives that block ovulation.

Serum thrombospondin-2 level changes with liver stiffness improvement in patients with type 2 diabetes.

OBJECTIVE: Baseline circulating thrombospondin-2 (TSP2) level was identified as a potential novel hepatic fibrosis biomarker that associates with development and progression of hepatic fibrosis in patients with nonalcoholic fatty liver disease and type 2 diabetes. Here, we investigated whether circulating TSP2 levels changed with improvement in liver stiffness (LS), which reflects liver fibrosis on transient elastography. DESIGN: Serum TSP2 levels were measured in participants from a randomized, open-label intervention study, at baseline and after 24-weeks treatment of either dapagliflozin 10 mg (N = 30) or sitagliptin 100 mg daily (N = 30). Vibration-controlled transient elastography was performed to evaluate the severity of hepatic fibrosis and steatosis using LS and controlled attenuation parameter (CAP), respectively. PATIENTS AND MEASUREMENTS: Among all 60 participants with similar clinical characteristics at baseline (mean HbA1c 8.9%, CAP 289 dB/m and LS 5.8 kPa), despite similar HbA1c lowering, treatment with dapagliflozin, but not sitagliptin, led to significant improvements in body weight (BW) (p = .012), CAP (p = .015) and LS (p = .011) after 24 weeks. RESULTS: Serum TSP2 level decreased significantly from baseline in dapagliflozin-treated participants (p = .035), whereas no significant change was observed with sitagliptin. In correlation analysis, change in serum TSP2 levels only positively correlated with change in LS (r = .487, p = .006), but not with changes in BW, CAP or HbA1c after dapagliflozin treatment. CONCLUSIONS: Serum TSP2 level decreased with LS after dapagliflozin treatment, and was independent of improvements in BW, glycemic control and hepatic steatosis, further supporting the potential of serum TSP2 level as a novel hepatic fibrosis biomarker in type 2 diabetes.

Effect of type 2 diabetes on the inducible degrader of LDL receptor.

The inducible degrader of LDL receptor (IDOL) acts as a post-transcriptional degrader of the LDL receptor (LDLR). IDOL is functionally active in the liver and in peripheral tissues. We have evaluated IDOL expression in circulating monocytes in subjects with and without type 2 diabetes and determined whether changes in IDOL expression could affect macrophage function like cytokine production in vitro. One hundred forty individuals with type 2 diabetes and 110 healthy control subjects were recruited. Cellular expression of IDOL and LDLR in peripheral blood CD14+ monocytes was measured by flow cytometry. The expression of intracellular IDOL was lower in individuals with diabetes than control (21.3 ± 4.6 mean fluorescence intensity × 1,000 vs. 23.8 ± 6.2, P < 0.01), and this was accompanied by an increase in cell surface LDLR (5.2 ± 3.0 mean fluorescence intensity × 1,000 vs. 4.3 ± 1.5, P < 0.01), LDL binding, and intracellular lipid (P < 0.01). IDOL expression correlated with HbA1c (r = -0.38, P < 0.01) and serum fibroblast growth factor-21 (FGF21) (r = -0.34, P < 0.01). Multivariable regression analysis, including age, sex, BMI, smoking, HbA1c, and log(FGF21), showed that HbA1c and FGF21 were significant independent determinants of IDOL expression. IDOL knockdown human monocyte-derived macrophages produced higher concentrations of interleukin 1 beta, interleukin 6, and TNFα than control macrophages upon stimulation with lipopolysaccharide (all P < 0.01). In conclusion, the expression of IDOL in CD14+ monocytes was decreased in type 2 diabetes and was associated with glycemia and serum FGF21 concentration.

RELA governs a network of islet-specific metabolic genes necessary for beta cell function.

AIMS/HYPOTHESIS: NF-κB activation unites metabolic and inflammatory responses in many diseases yet less is known about the role that NF-κB plays in normal metabolism. In this study we investigated how RELA impacts the beta cell transcriptional landscape and provides network control over glucoregulation. METHODS: We generated novel mouse lines harbouring beta cell-specific deletion of either the Rela gene, encoding the canonical NF-κB transcription factor p65 (ßp65KO mice), or the Ikbkg gene, encoding the NF-κB essential modulator NEMO (ßNEMOKO mice), as well as ßA20Tg mice that carry beta cell-specific and forced transgenic expression of the NF-κB-negative regulator gene Tnfaip3, which encodes the A20 protein. Mouse studies were complemented by bioinformatics analysis of human islet chromatin accessibility (assay for transposase-accessible chromatin with sequencing [ATAC-seq]), promoter capture Hi-C (pcHi-C) and p65 binding (chromatin immunoprecipitation-sequencing [ChIP-seq]) data to investigate genome-wide control of the human beta cell metabolic programme. RESULTS: Rela deficiency resulted in complete loss of stimulus-dependent inflammatory gene upregulation, consistent with its known role in governing inflammation. However, Rela deletion also rendered mice glucose intolerant because of functional loss of insulin secretion. Glucose intolerance was intrinsic to beta cells as ßp65KO islets failed to secrete insulin ex vivo in response to a glucose challenge and were unable to restore metabolic control when transplanted into secondary chemical-induced hyperglycaemic recipients. Maintenance of glucose tolerance required Rela but was independent of classical NF-κB inflammatory cascades, as blocking NF-κB signalling in vivo by beta cell knockout of Ikbkg (NEMO), or beta cell overexpression of Tnfaip3 (A20), did not cause severe glucose intolerance. Thus, basal p65 activity has an essential and islet-intrinsic role in maintaining normal glucose homeostasis. Genome-wide bioinformatic mapping revealed the presence of p65 binding sites in the promoter regions of specific metabolic genes and in the majority of islet enhancer hubs (~70% of ~1300 hubs), which are responsible for shaping beta cell type-specific gene expression programmes. Indeed, the islet-specific metabolic genes Slc2a2, Capn9 and Pfkm identified within the large network of islet enhancer hub genes showed dysregulated expression in ßp65KO islets. CONCLUSIONS/INTERPRETATION: These data demonstrate an unappreciated role for RELA as a regulator of islet-specific transcriptional programmes necessary for the maintenance of healthy glucose metabolism. These findings have clinical implications for the use of anti-inflammatories, which influence NF-κB activation and are associated with diabetes.

PCTAIRE Protein Kinase 1 (PCTK1) Suppresses Proliferation, Stemness, and Chemoresistance in Colorectal Cancer through the BMPR1B-Smad1/5/8 Signaling Pathway.

Colorectal cancer (CRC) is the third most common cancer and a leading cause of cancer-related mortality worldwide. Even with advances in therapy, CRC mortality remains high. Therefore, there is an urgent need to develop effective therapeutics for CRC. PCTAIRE protein kinase 1 (PCTK1) is an atypical member of the cyclin-dependent kinase (CDK) family, and the function of PCTK1 in CRC is poorly understood. In this study, we found that patients with elevated PCTK1 levels had a better overall survival rate in CRC based on the TCGA dataset. Functional analysis also showed that PCTK1 suppressed cancer stemness and cell proliferation by using PCTK1 knockdown (PCTK1-KD) or knockout (PCTK1-KO) and PCTK1 overexpression (PCTK1-over) CRC cell lines. Furthermore, overexpression of PCTK1 decreased xenograft tumor growth and knockout of PCTK1 significantly increased in vivo tumor growth. Moreover, knockout of PCTK1 was observed to increase the resistance of CRC cells to both irinotecan (CPT-11) alone and in combination with 5-fluorouracil (5-FU). Additionally, the fold change of the anti-apoptotic molecules (Bcl-2 and Bcl-xL) and the proapoptotic molecules (Bax, c-PARP, p53, and c-caspase3) was reflected in the chemoresistance of PCTK1-KO CRC cells. PCTK1 signaling in the regulation of cancer progression and chemoresponse was analyzed using RNA sequencing and gene set enrichment analysis (GSEA). Furthermore, PCTK1 and Bone Morphogenetic Protein Receptor Type 1B (BMPR1B) in CRC tumors were negatively correlated in CRC patients from the Timer2.0 and cBioPortal database. We also found that BMPR1B was negatively correlated with PCTK1 in CRC cells, and BMPR1B expression was upregulated in PCTK1-KO cells and xenograft tumor tissues. Finally, BMPR1B-KD partially reversed cell proliferation, cancer stemness, and chemoresistance in PCTK1-KO cells. Moreover, the nuclear translocation of Smad1/5/8, a downstream molecule of BMPR1B, was increased in PCTK1-KO cells. Pharmacological inhibition of Smad1/5/8 also suppressed the malignant progression of CRC. Taken together, our results indicated that PCTK1 suppresses proliferation and cancer stemness and increases the chemoresponse of CRC through the BMPR1B-Smad1/5/8 signaling pathway.

Improved Productivity Using Deep Learning-assisted Reporting for Lumbar Spine MRI.

Background Lumbar spine MRI studies are widely used for back pain assessment. Interpretation involves grading lumbar spinal stenosis, which is repetitive and time consuming. Deep learning (DL) could provide faster and more consistent interpretation. Purpose To assess the speed and interobserver agreement of radiologists for reporting lumbar spinal stenosis with and without DL assistance. Materials and Methods In this retrospective study, a DL model designed to assist radiologists in the interpretation of spinal canal, lateral recess, and neural foraminal stenoses on lumbar spine MRI scans was used. Randomly selected lumbar spine MRI studies obtained in patients with back pain who were 18 years and older over a 3-year period, from September 2015 to September 2018, were included in an internal test data set. Studies with instrumentation and scoliosis were excluded. Eight radiologists, each with 2-13 years of experience in spine MRI interpretation, reviewed studies with and without DL model assistance with a 1-month washout period. Time to diagnosis (in seconds) and interobserver agreement (using Gwet κ) were assessed for stenosis grading for each radiologist with and without the DL model and compared with test data set labels provided by an external musculoskeletal radiologist (with 32 years of experience) as the reference standard. Results Overall, 444 images in 25 patients (mean age, 51 years ± 20 [SD]; 14 women) were evaluated in a test data set. DL-assisted radiologists had a reduced interpretation time per spine MRI study, from a mean of 124-274 seconds (SD, 25-88 seconds) to 47-71 seconds (SD, 24-29 seconds) (P < .001). DL-assisted radiologists had either superior or equivalent interobserver agreement for all stenosis gradings compared with unassisted radiologists. DL-assisted general and in-training radiologists improved their interobserver agreement for four-class neural foraminal stenosis, with κ values of 0.71 and 0.70 (with DL) versus 0.39 and 0.39 (without DL), respectively (both P < .001). Conclusion Radiologists who were assisted by deep learning for interpretation of lumbar spinal stenosis on MRI scans showed a marked reduction in reporting time and superior or equivalent interobserver agreement for all stenosis gradings compared with radiologists who were unassisted by deep learning. © RSNA, 2022 Online supplemental material is available for this article. See also the editorial by Hayashi in this issue.

Impact of contrast-enhanced mammography in surgical management of breast cancers for women with dense breasts: a dual-center, multi-disciplinary study in Asia.

OBJECTIVE: To evaluate the impact of pre-operative contrast-enhanced mammography (CEM) in breast cancer patients with dense breasts. METHODS: We conducted a retrospective review of 232 histologically proven breast cancers in 200 women (mean age: 53.4 years ± 10.2) who underwent pre-surgical CEM imaging across two Asian institutions (Singapore and Taiwan). Majority (95.5%) of patients had dense breast tissue (BI-RADS category C or D). Surgical decision was recorded in a simulated blinded multi-disciplinary team setting on two separate scenarios: (i) pre-CEM setting with standard imaging, and clinical and histopathological results; and (ii) post-CEM setting with new imaging and corresponding histological findings from CEM. Alterations in surgical plan (if any) because of CEM imaging were recorded. Predictors CEM of patients who benefitted from surgical plan alterations were evaluated using logistic regression. RESULTS: CEM resulted in altered surgical plans in 36 (18%) of 200 patients in this study. CEM discovered clinically significant larger tumor size or extent in 24 (12%) patients and additional tumors in 12 (6%) patients. CEM also detected additional benign/false-positive lesions in 13 (6.5%) of the 200 patients. Significant predictors of patients who benefitted from surgical alterations found on multivariate analysis were pre-CEM surgical decision for upfront breast conservation (OR, 7.7; 95% CI, 1.9-32.1; p = 0.005), architectural distortion on mammograms (OR, 7.6; 95% CI, 1.3-42.9; p = .022), and tumor size of ≥ 1.5 cm (OR, 1.5; 95% CI, 1.0-2.2; p = .034). CONCLUSION: CEM is an effective imaging technique for pre-surgical planning for Asian breast cancer patients with dense breasts. KEY POINTS: • CEM significantly altered surgical plans in 18% (nearly 1 in 5) of this Asian study cohort with dense breasts. • Significant patient and imaging predictors for surgical plan alteration include (i) patients considered for upfront breast-conserving surgery; (ii) architectural distortion lesions; and (iii) tumor size of ≥ 1.5 cm. • Additional false-positive/benign lesions detected through CEM were uncommon, affecting only 6.5% of the study cohort.

Using comprehensive geriatric assessment for older adults undertaking a facility-based transition care program to evaluate functional outcomes: a feasibility study.

BACKGROUND: The study aimed to evaluate the feasibility of using a comprehensive geriatric assessment (CGA) in a residential transition care setting to measure older adults' functional outcomes. METHODS: A convenience sample of older adults (n = 10) and staff (n = 4) was recruited. The feasibility of using assessment tools that comprise a CGA to comprehensively measure function in physical, cognitive, social and emotional domains was evaluated pre- and post-rehabilitation. RESULTS: 10 older adults (mean ± SD age = 78.9 ± 9.1, n = 6 male) completed a CGA performed using assessments across physical, cognitive, social and emotional domains. The CGA took 55.9 ± 7.3 min to complete. Staff found CGA using the selected assessment tools to be acceptable and suitable for the transition care population. Older adults found the procedure to be timely and 60% found the assessments easy to comprehend. Participating in CGA also assisted older adults in understanding their present state of health. The older adults demonstrated improvements across all assessed domains including functional mobility (de Morton Mobility Index; baseline 41.5 ± 23.0, discharge 55.0 ± 24.0, p = 0.01) and quality of life (EQ-5D-5L; baseline 59.0 ± 21.7, discharge 78.0 ± 16.0, p < 0.01). CONCLUSIONS: Incorporating CGA to evaluate functional outcomes in transition care using a suite of assessment tools was feasible and enabled a holistic assessment.

Pediatric non-alcoholic fatty liver disease predicted by anthropometric values through transient elastography.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is the leading cause of pediatric chronic liver disease, which is strongly associated with obesity. Transient elastography, together with anthropometric values including waist-to-height ratio (WHtR) and body mass index (BMI) z-scores are a more precise diagnostic method of NAFLD than ultrasonography. Through transient elastography, we investigate the principal anthropometric values associated with pediatric NAFLD. METHODS: Healthy children between the ages of 6-18 years whose BMIs were ≥85% of normal were recruited as the overweight-and-obese group, and children whose BMIs ranged between 5%-85% were recruited as the control group. Non-alcoholic fatty liver disease was evaluated via transient elastography. BMI z-score and WHtR were measured. RESULTS: A total of 107 (58 overweight-and-obese, 49 control) children were recruited. As evaluated by transient elastography, children in the overweight-and-obese group had significantly higher controlled attenuation parameter and liver stiffness measurement values than the control group. To detect fatty liver, WHtR with a cut-off point of 0.481 and BMI z-score with cut-off point of 1.075 had the best sensitivity and specificity. To identify liver stiffness or inflammation, WHtR with cut-off point of 0.514 and BMI z-score with cut-off point of 1.62 had the best sensitivity and specificity. Controlled attenuation parameter demonstrated a fair correlation with WHtR and BMI z-scores, even in the normal range of these parameters. CONCLUSIONS: Transient elastography together with anthropometric measurements demonstrate that pediatric NAFLD may develop earlier than expected. We present principal anthropometric values associated with pediatric NAFLD.

Optimization of Blood Handling and Peripheral Blood Mononuclear Cell Cryopreservation of Low Cell Number Samples.

BACKGROUND: Rural/remote blood collection can cause delays in processing, reducing PBMC number, viability, cell composition and function. To mitigate these impacts, blood was stored at 4 °C prior to processing. Viable cell number, viability, immune phenotype, and Interferon-γ (IFN-γ) release were measured. Furthermore, the lowest protective volume of cryopreservation media and cell concentration was investigated. METHODS: Blood from 10 individuals was stored for up to 10 days. Flow cytometry and IFN-γ ELISPOT were used to measure immune phenotype and function on thawed PBMC. Additionally, PBMC were cryopreserved in volumes ranging from 500 µL to 25 µL and concentration from 10 × 106 cells/mL to 1.67 × 106 cells/mL. RESULTS: PBMC viability and viable cell number significantly reduced over time compared with samples processed immediately, except when stored for 24 h at RT. Monocytes and NK cells significantly reduced over time regardless of storage temperature. Samples with >24 h of RT storage had an increased proportion in Low-Density Neutrophils and T cells compared with samples stored at 4 °C. IFN-γ release was reduced after 24 h of storage, however not in samples stored at 4 °C for >24 h. The lowest protective volume identified was 150 µL with the lowest density of 6.67 × 106 cells/mL. CONCLUSION: A sample delay of 24 h at RT does not impact the viability and total viable cell numbers. When long-term delays exist (>4 d) total viable cell number and cell viability losses are reduced in samples stored at 4 °C. Immune phenotype and function are slightly altered after 24 h of storage, further impacts of storage are reduced in samples stored at 4 °C.

ASTEROID stereotest v1.0: lower stereo thresholds using smaller, denser and faster dots.

PURPOSE: In 2019, we described ASTEROID, a new stereotest run on a 3D tablet computer which involves a four-alternative disparity detection task on a dynamic random-dot stereogram. Stereo thresholds measured with ASTEROID were well correlated with, but systematically higher than (by a factor of around 1.5), thresholds measured with previous laboratory stereotests or the Randot Preschool clinical stereotest. We speculated that this might be due to the relatively large, sparse dots used in ASTEROID v0.9. Here, we introduce and test the stereo thresholds and test-repeatability of the new ASTEROID v1.0, which uses precomputed images to allow stereograms made up of much smaller, denser dots. METHODS: Stereo thresholds and test/retest repeatability were tested and compared between the old and new versions of ASTEROID (n = 75) and the Randot Circles (n = 31) stereotest, in healthy young adults. RESULTS: Thresholds on ASTEROID v1.0 are lower (better) than on ASTEROID v0.9 by a factor of 1.4, and do not differ significantly from thresholds on the Randot Circles. Thresholds were roughly log-normally distributed with a mean of 1.54 log10 arcsec (35 arcsec) on ASTEROID v1.0 compared to 1.70 log10 arcsec (50 arcsec) on ASTEROID v0.9. The standard deviation between observers was the same for both versions, 0.32 log10 arcsec, corresponding to a factor of 2 above and below the mean. There was no difference between the versions in their test/retest repeatability, with 95% coefficient of repeatability = 0.46 log10 arcsec (a factor of 2.9 or 1.5 octaves) and a Pearson correlation of 0.8 (comparable to other clinical stereotests). CONCLUSION: The poorer stereo thresholds previously reported with ASTEROID v0.9 appear to have been due to the relatively large, coarse dots and low density used, rather than to some other aspect of the technology. Employing the small dots and high density used in ASTEROID v1.0, thresholds and test/retest repeatability are similar to other clinical stereotests.

Galectin-3 and risk of cardiovascular events and all-cause mortality in type 2 diabetes.

AIMS: Recent clinical studies have shown that galectin-3 is a prognostic indicator in patients with coronary heart disease and in patients with heart failure. Experimental data suggest that galectin-3 may play a role in atherogenesis. We have evaluated whether serum galectin-3 level is associated with cardiovascular outcome in type 2 diabetes. MATERIALS AND METHODS: Galectin-3 was measured in baseline samples in 1495 persons with type 2 diabetes. The primary cardiovascular outcome, incident cardiovascular events, was defined as first non-fatal myocardial infarction, non-fatal stroke, coronary revascularization, or death from cardiovascular cause. The secondary outcome was all-cause mortality. RESULTS: At baseline, 12% of the subjects had prevalent cardiovascular disease. Serum galectin-3 was increased in the group with incident cardiovascular events compared with those who remained free of events during follow up (9.03 ± 2.98 ng/mL vs 8.15 ± 2.76, P < 0.01). Serum galectin-3 was also significantly increased in those subjects with a fatal outcome. The hazard ratios (HR) for cardiovascular events and all-cause mortality for individuals in the top quartile were 2.50 (95% CI 1.87, 3.36, P < 0.001) and 3.92 (95%CI 2.55, 6.01, P < 0.001), respectively. In a multivariate Cox regression analysis including traditional risk factors, log (eGFR), baseline albuminuria, and cardiovascular disease status, the HR per standard deviation change in galectin-3 was 1.13 (95% CI 1.02, 1.26, P = 0.02) for cardiovascular events and 1.17 (95% CI 1.01, 1.35, P = 0.04) for all-cause mortality. CONCLUSIONS: Serum galectin-3 is associated with adverse cardiovascular outcomes in persons with type 2 diabetes independent of traditional risk factors.

A sclerocornea-associated RAD21 variant induces corneal stroma disorganization.

Sclerocornea is a cornea opacification disorder. Disorganized corneal stroma fibrils are observed in patients' cornea. Previously we identified a RAD21C1348T variant that is associated with a peripheral sclerocornea pedigree. To explore whether this RAD21 variant can induce sclerocornea-related phenotype, and to investigate the possible mechanisms of such phenotype, the orthologous rad21 wild-type and variant mRNAs were injected into Xenopus laevis embryos and the developed eyes were subjected for histological examination. Transmission electron microscopy was applied for corneal stroma organization check. rad21 is highly expressed in the eye region during X. laevis development. Disrupted eye development was observed in the rad21 variant injected embryos. Disorganized corneal stroma and decreased diameters of collagen fibrils were observed in the rad21 variant injected X. laevis eyes. These eye defects can be rescued by overexpression of the wild-type rad21. Histological examination found stroma attracting center, a key structure in X. laevis corneal development, was impaired in rad21 variant injected embryos. Our results suggest a key role of RAD21 during corneal development. Our data indicates the RAD21R450C variant contributes to peripheral sclerocornea by disturbing collagen fibril organization in the corneal stroma.

Does oral Nonsteroidal Anti-inflammatory Drugs (NSAIDs) premedication in patients with irreversible pulpitis increase the success rate of inferior alveolar nerve block?

Data sources PubMed, EBSCOhost and Scopus database up till 9 September 2017 in English language. Clinical trial registry and reference lists of published systematic reviews, textbooks and selected articles were also searched.Study selection Population-based randomised clinical trials comparing effects of any single NSAID versus placebo as an oral premedication on the efficacy of IANB in achieving anaesthesia in patients ranging from age 14-68 with irreversible pulpitis who were undergoing nonsurgical root canal therapy in mandibular posterior teeth.Data extraction and synthesis Studies were selected, reviewed and extracted by two independent reviewers using a standardised extraction form. They assessed risk of bias using the revised Cochrane Risk of Bias Tool for Randomised Trials (ROB 2.0). Meta-analysis was performed using a random effects model. I2 was used to evaluate heterogeneity. A subgroup analysis was conducted to investigate the dose-response effect of ibuprofen. Sensitivity analyses were performed by restricting studies with a low risk of bias and by using a fixed effects model (using STATA 14.1 software). Publication bias was assessed using funnel plot asymmetry and Egger regression tests. Trial Sequential Analysis (TSA) was conducted to assess the risk of random errors and to determine the required sample size, which helped to evaluate if evidence is conclusive. Quality of evidence was rated by GRADE pro GDT software.Results Thirteen randomised clinical studies that investigated the efficacy of oral premedication (30-60 minutes prior) with a single NSAID in promoting the anaesthetic success of IANBs involving 1,174 patients between the age of 14 to 68 were included. Of these, 1,034 were evaluated as a comparison to placebo while the remainder were comparisons to various medications.Among the 13 RCTs, eight showed a low risk of bias, four trials showed a high risk and the remaining one showed unclear risk. For all studies, intervention provision and outcome measurement were performed at the same visit with no follow-up periods. Of the 1,034 participants, 493 (47.6%) reported successful anaesthetic outcomes. Quantitative pooling of the results showed that the use of any NSAID significantly increased the anaesthetic success of IANB compared with placebo (RR=1.96; 95% CI, 1.63-2.35; I2= 6.8%).Subgroup analysis of nine trials showed that ibuprofen, diclofenac 50mg and ketorolac 10mg had a statistically significantly effect in increasing the anaesthetic success of IANBs compared with placebo (RR= 1.83, 95% CI, 1.43-2.35, I2= 20.8%; RR= 2.56, 95% CI, 1.46-4.50, I2= 44.8%; and RR= 2.07, 95% CI, 1.47-2.90, I2=0% respectively). While Ibuprofen >400mg was significantly more effective than placebo (RR= 1.85; 95% CI, 1.39-2.45; I2=26.7%), ibuprofen <400mg showed no statistical association (RR=1.78; 95% CI, 0.90-3.55; I2=38.70%). Funnel plot showed low publication bias. GRADE evaluation showed that the accumulated evidence for premedication with NSAIDs demonstrated high quality. I2 results showed a low heterogeneity among trials. TSA concluded that the results of the meta-analyses showing premedication with NSAIDs increased the success rate of IANB anaesthesia were valid.Conclusions High quality evidence demonstrated that the oral premedication with NSAIDs increases the success rate of IANB in patients with irreversible pulpitis.

Galectin-3 is independently associated with progression of nephropathy in type 2 diabetes mellitus.

AIMS/HYPOTHESIS: Galectin-3 has been implicated in cardiac and renal fibrosis and serves as a prognostic clinical indicator in heart failure. The aim of the present study was to evaluate whether serum galectin-3 level is associated with progressive kidney disease in type 2 diabetes. METHODS: Galectin-3 was measured in baseline samples by ELISA in 1320 participants with type 2 diabetes with eGFR ≥30 ml min-1 1.73 m-2. The primary outcome was defined as doubling of serum creatinine and/or initiation of renal replacement therapy during follow-up. The secondary outcome was progression to macroalbuminuria in individuals with normo- or microalbuminuria at baseline. RESULTS: Serum galectin-3 levels were significantly increased in a random subgroup of 270 type 2 diabetic individuals with eGFR >60 ml min-1 1.73 m-2 compared with an age- and sex-matched non-diabetic control group (7.58 ± 2.29 ng/ml vs 6.10 ± 1.91 ng/ml, respectively, p < 0.01). In the whole diabetic cohort, after a mean follow-up of 9 years, galectin-3 was independently associated with doubling of serum creatinine (HR 1.19; 95% CI 1.14, 1.24, p < 0.001) and incident macroalbuminuria (HR 1.20; 95% CI 1.12, 1.30, p < 0.001), even after adjusting for traditional risk factors, baseline eGFR and albuminuria status. Individuals with galectin-3 levels in the highest quartile had a fourfold risk of renal function loss and threefold risk of incident macroalbuminuria. CONCLUSIONS/INTERPRETATION: Serum galectin-3 was independently associated with progressive renal disease in type 2 diabetes. Further mechanistic studies are warranted to determine whether galectin-3 is simply a disease biomarker or is also a mediator of the development and progression of diabetic nephropathy.

Improving the quality of electroretinogram recordings using active electrodes.

The aim of this study was to compare the quality of electroretinogram (ERG) recordings using a custom built active electrode with attached amplifier versus a standard (passive) ERG electrode. Scotopic and photopic ERG responses were recorded from five adult albino rabbits using a custom built active electrode on one eye and a passive electrode on the other. For the active electrode, the ERG-jet electrode (Universo S.A., La Chaux-De-Fonds, Switzerland) was used as the transducer with the cable cut short and soldered directly to the input of a customized amplifier. The passive electrode was a standard ERG jet electrode. The signal to noise ratio and reproducibility of ERGs were compared. The noise was significantly lower in the active electrode compared to the passive electrode (p = 0.009) resulting in signals being recorded at lower stimulation strengths with the active electrode. The scotopic a-wave was significantly larger in the active electrode at all supra-threshold stimulation intensities (p < 0.05) and the scotopic b-wave amplitudes were also higher in the active electrode at all supra-threshold stimulation intensities but was only statistically significant between -3.25 and -1 log cd.s.m-2 (p < 0.05). The photopic a- and b-wave amplitudes were also higher in the active electrode and statistically significant between -0.75 and 0.48 log cd.s.m-2 for the a-wave and -1.25 to -1 log cd.s.m-2 for the b-wave (p < 0.05). The intra-observer repeatability, inter-sessions reproducibility and reliability of the signals were better in the active electrode as evidenced by lower coefficient of variation (CV) and coefficient of repeatability (CR) with high intra-class correlation coefficient (ICC) of the a- and b-wave parameters of the active electrode. These findings suggest that the custom built active ERG electrode produces less noise than the passive electrode, allowing responses to be recorded at lower stimulation strengths. It produces greater signal amplitudes and improved reproducibility and is therefore a better device for investigating retinal function.

How cognitive engagement fluctuates during a team-based learning session and how it predicts academic achievement.

The objective of the paper is to report findings of two studies that attempted to find answers to the following questions: (1) What are the levels of cognitive engagement in TBL? (2) Are there differences between students who were more exposed to TBL than students who were less exposed to TBL? (3) To which extent does cognitive engagement fluctuate as a function of the different activities involved in TBL? And (4) How do cognitive engagement scores collected over time correlate with each other and with academic achievement? The studies were conducted with Year-1 and -2 medical students enrolled in a TBL curriculum (N = 175, 62 female). In both studies, six measurements of cognitive engagement were taken during the distinct TBL activities (preparation phase, individual/team readiness assurance test, burning questions, and application exercises). Data were analysed by means of one-way repeated-measures ANOVAs and path modelling. The results of the repeated-measures ANOVA revealed that cognitive engagement systematically fluctuated as a function of the distinct TBL activities. In addition, Year-1 students reported significantly higher levels of cognitive engagement compared to Year-2 students. Finally, cognitive engagement was a significant predictor of performance (ß = .35). The studies presented in this paper are a first attempt to relate the different activities undertaken in TBL with the extent to which they arouse cognitive engagement with the task at hand. Implications of these findings for TBL are discussed.

Caries removal using lasers.

Data sourcesCochrane Oral Health Trials Register, Cochrane Central Register of Controlled Trials (CENTRAL), Medline Ovid, Embase Ovid, ProQuest Dissertations and Theses, Zetoc, ISI Web of Knowledge, US National Institutes of Health Ongoing Trials Register ClinicalTrials.gov, and the WHO International Clinical Trials Registry Platform for ongoing trials up to June 2016.Study selectionRandomised controlled trials, split-mouth trials and cluster-randomised trials comparing laser ablation to drill removal of caries with no restriction in language or participants' age.Data extraction and synthesisStudies were selected and reviewed independently by two reviewers and standard data items extracted. The reviewers assessed the risk of bias of all studies using Cochrane's 'Risk of bias' tool. For all dichotomous outcomes, risk ratios (RR) with 95% of confidence intervals (CI) were calculated, and mean difference (MD) was calculated for continuous data. Primary outcomes were effectiveness in caries excavation and reports of pain. Data were combined using random-effects models. Main results for caries removal were summarised and presented using GRADE proGDT software. Meta-analyses were performed on studies that reported quantitative data.ResultsSix split-mouth randomised controlled trials and three parallel-group randomised trials involving 1,498 primary and permanent teeth from 662 participants ranging from 3.5 to 84 years old were included. Seven hundred and seventy-seven teeth were treated with laser only, 732 with mechanical drills only, and 12 teeth were treated with both techniques in the same tooth on separate caries.Only four studies evaluated caries removal. Of those four, only two reported quantitative data. After meta-analysis, the results showed no significant difference in effectiveness of caries removal between the two treatment methods ((RR) 1.00, 95% (CI) 0.99 to 1.01). Of the five studies that assessed pain, three studies using five-point or six-point pain scale reported less pain experienced using laser. Two studies did not provide complete data for analysis. As for the secondary outcomes of marginal integrity of restorations (three studies), durability (four studies), recurrent caries (two studies), pulpal inflammation or necrosis (four studies), overall results showed no evidence of a difference. When considering the need for anaesthesia (four studies), and participant discomfort (five studies), the overall results showed that the need for anaesthesia and participant discomfort was lower with the laser treatment. Interestingly, the one study that reported operator preference showed that dentists preferred conventional preparation method over laser (P<0.001).ConclusionsThere was insufficient evidence to show that laser removal of caries was more or less efficient than traditional mechanical technique. However, there was some low quality evidence in favour of laser therapy for pain control, need for anaesthesia and patient comfort.

Escherichia coli Topoisomerase IV E Subunit and an Inhibitor Binding Mode Revealed by NMR Spectroscopy.

Bacterial topoisomerases are attractive antibacterial drug targets because of their importance in bacterial growth and low homology with other human topoisomerases. Structure-based drug design has been a proven approach of efficiently developing new antibiotics against these targets. Past studies have focused on developing lead compounds against the ATP binding pockets of both DNA gyrase and topoisomerase IV. A detailed understanding of the interactions between ligand and target in a solution state will provide valuable information for further developing drugs against topoisomerase IV targets. Here we describe a detailed characterization of a known potent inhibitor containing a 9H-pyrimido[4,5-b]indole scaffold against the N-terminal domain of the topoisomerase IV E subunit from Escherichia coli (eParE). Using a series of biophysical and biochemical experiments, it has been demonstrated that this inhibitor forms a tight complex with eParE. NMR studies revealed the exact protein residues responsible for inhibitor binding. Through comparative studies of two inhibitors of markedly varied potencies, it is hypothesized that gaining molecular interactions with residues in the α4 and residues close to the loop of ß1-α2 and residues in the loop of ß3-ß4 might improve the inhibitor potency.