Tamoxifen-Independent Recombination in the RIP-CreER Mouse

The inducible Cre-lox system is a valuable tool to study gene function in a spatial and time restricted fashion in mouse models. This strategy relies on the limited background activity of the modified Cre recombinase (CreER) in the absence of its inducer, the competitive estrogen receptor ligand, tamoxifen. The RIP-CreER mouse (Tg (Ins2-cre/Esr1) 1Dam) is among the few available β-cell specific CreER mouse lines and thus it has been often used to manipulate gene expression in the insulin-producing cells of the endocrine pancreas

Characterization of the SKA1-Low prototype station Aperture Array Verification System 2

The low frequency component of the Square Kilometre Array (SKA1-Low) will be an aperture phased array located at the Murchison Radio-astronomy Observatory (MRO) site in Western Australia. It will be composed of 512 stations, each consisting of 256 log-periodic dual-polarized antennas, and will operate in the low frequency range (50 to 350 MHz) of the SKA bandwidth. The Aperture Array Verification System 2 (AAVS2), operational since late 2019, is the last full-size engineering prototype station deployed at the MRO site before the start of the SKA1-Low construction phase. The aim of this paper is to characterize the station performance through commissioning observations at six different frequencies (55, 70, 110, 160, 230, and 320 MHz) collected during its first year of activities. We describe the calibration procedure, present the resulting all-sky images and their analysis, and discuss the station calibratability and system stability. Using the difference imaging method, we also derive estimates of the SKA1-Low sensitivity for the same frequencies and compare them with those obtained through electromagnetic simulations across the entire telescope bandwidth, finding good agreement (within 13%). Moreover, our estimates exceed the SKA1-Low requirements at all considered frequencies by up to a factor of ∼2.3. Our results are very promising and allow for an initial validation of the AAVS2 prototype station performance, which is an important step toward the coming SKA1-Low telescope construction and science

Report of Two Cases of Pediatric IgG4-Related Lymphadenopathy (IgG4-LAD): IgG4-Related Disease (IgG4-RD) or a Distinct Clinical Pathological Entity?

IgG4-related disease (IgG4-RD) is a recently discovered immune-mediated fibroinflammatory condition, uncommon in the pediatric population, that could involve multiple organs and induce cancer-like lesions and organ damage. Its main features are multiple injuries in different sites, a dense lymphoplasmacytic infiltrate rich in IgG4 plasma cells, storiform fibrosis, and often high serological concentrations of IgG4. Autoimmune pancreatitis is the most common manifestation, mainly in adults. Two cases of IgG4-RD in children with lymph node localization of disease are reported. Localized or systemic lymph node involvement is common, but lymph node enlargement as the first and only manifestation of IgG4-RD is unusual, and therefore, hard to differentiate from other diseases. IgG4-related lymphadenopathy (IgG4-LAD) is most likely a distinct disease, described as isolated lymphadenopathy, related to the presence of elevated numbers of IgG4-positive plasma cells. Both disorders are likely to be misdiagnosed in children because they are characterized by rare and polymorphic features. IgG4-RD and IgG4-LAD should be considered in the differential diagnosis of disorders characterized by lymphadenopathy of uncertain etiology

β<i>-galactosidase</i> gene (<i>lacZ</i>) expression in <i>Pdx1-CreER, Rosa26-lacZ</i> mice.

<p><b>A-C.</b> LacZ staining of the pancreatic tissue from 4-month-old tamoxifen-treated <i>Pdx1-CreER, Rosa26-lacZ</i> mice (A), and <i>Rosa26-lacZ</i> (B) and untreated <i>Pdx1-CreER, Rosa26-lacZ</i> (C) mice. <b>D.</b> Quantification and statistical analysis of the lacZ-positive area relative to the total islet area in tamoxifen-treated or untreated <i>Pdx1-CreER, Rosa26-lacZ</i> mice, and tamoxifen-treated <i>Rosa26-lacZ</i> mice. Dots represent individual islets from 3 mice in each genotype group. The p value of a t-test between groups is shown.</p

β<i>-galactosidase</i> gene (<i>lacZ</i>) expression in <i>RIP-CreER, Rosa26-lacZ</i> mice.

<p><b>A-C.</b> LacZ staining of the pancreatic tissue from 10-week-old tamoxifen-treated <i>RIP-CreER, Rosa26-lacZ</i> mice (A), and <i>Rosa26-lacZ</i> (B) and untreated <i>RIP-CreER, Rosa26-lacZ</i> (C) mice. <b>D.</b> Quantification and statistical analysis of the lacZ-positive area relative to the total islet area in tamoxifen-treated or untreated <i>RIP-CreER, Rosa26-lacZ</i> mice, and tamoxifen-treated <i>Rosa26-lacZ</i> mice. Dots represent individual islets from 3 mice in each genotype group. The p value of a t-test between groups is shown.</p

Tamoxifen-independent knockout of <i>PTBP1</i> in adult β-cells of <i>RIP-CreER, PTBP1^loxP/loxP</i> mice.

<p><b>A-L</b>. Confocal microscopy images of pancreatic cryosections from 8-week-old tamoxifen-treated <i>RIP-CreER, PTBP1^loxP/loxP</i> (A-D) and <i>PTBP1^loxP/loxP</i> (E-H) and untreated <i>RIP-CreER, PTBP1^loxP/loxP</i> (I-L) mice stained with DAPI (A, E and I), as well as with anti-PTBP1 (B, F and J) and anti-insulin antibodies (C, G and K). Triple stainings (merge) are shown in D, H and L. Scale bars in D, H and L equal 20 µm. <b>M.</b> Quantification and statistical analysis of PTBP1-positive cells relative to total insulin-positive cells in tamoxifen-treated and untreated <i>RIP-CreER</i>, <i>PTBP1^loxP/loxP</i> mice, and tamoxifen-treated <i>PTBP1^loxP/loxP</i> mice. Dots represent individual islets from 4 mice in each genotype group. The p value of a t-test between groups is shown.</p

Tamoxifen-independent expression of <i>HA3-ICA512-CCF</i> in β-cells of <i>RIP-CreER, R26-Stop-HA3-ICA512-CCF</i> mice.

<p><b>A</b>. Strategy for the generation of the inducible <i>R26-Stop-HA3-ICA512-CCF</i> knock-in mouse line. <b>B</b>. Western blotting of pancreatic islet extracts from 3-month-old tamoxifen-treated or untreated <i>RIP-CreER, R26-Stop-HA3-ICA512-CCF</i> mice or tamoxifen-treated <i>R26-Stop-HA3-ICA512-CCF</i> mice with anti-HA and anti-γ-tubulin antibodies (similar results were obtained from 3 independent experiments). <b>C-N</b>. Confocal microscopy images of pancreatic cryosections from 4-month-old tamoxifen-treated <i>RIP-CreER, R26-Stop-HA3-ICA512-CCF</i> (C-F) and <i>R26-Stop-HA3-ICA512-CCF</i> (G-J) and untreated <i>RIP-CreER, R26-Stop-HA3-ICA512-CCF</i> (K-N) mice stained with DAPI (C, G and K), as well as with anti-insulin (D, H and L) and anti-HA antibodies (E, I and M). Triple stainings (merge) are shown in F, J and N. Scale bars in F, J and N equal 10 µm. <b>O.</b> Quantification and statistical analysis of HA3-ICA512-CCF-positive cells relative to total insulin-positive cells in tamoxifen-treated or untreated <i>RIP-CreER, R26-Stop-HA3-ICA512-CCF</i> mice, and tamoxifen-treated <i>R26-Stop-HA3-ICA512-CCF</i> mice. Dots represent individual islets from 3 mice in each genotype group. The p value of a t-test between groups is shown.</p

The percutaneous treatment of Patent Foramen Ovale, an effective and safe therapeutic choice

Introduction: The aim of our study is to evaluate the feasibility, safety and efficacy of the percutaneous closure of PFO (abnormal communication between the right and left atrium). Methods: Between July 2009 and October 2012 percutaneous closure was performed in 37 patients. The presence of PFO was diagnosed through the use of ultrasound techniques: transcranial doppler with contrast (cTCD), transthoracic echocardiography(TTE) and transesophageal echocardiography (TEE). Follow-up was composed consisted of a Holter ECG 7 days after the closure with a 24 hour heart rhythm monitoring, to evaluate eventual arrhythmia cases and programmed controls which included a TTE at 1-3 months, TTE+ cTCD at 6-12 months, to evaluate the right positioning of the device and the complete closure of the defect. Results: We obtained 100% of procedural success (correct and stable implantation of the device in a perfect position on the interatrial septum).No complications were recorded during the procedure and no new onset atrial fibrillation was detected in any patients after the PFO closure. The follow up with cTCD and TEE reported a closing rate of 86.7%. No new clinical cerebrovascular events occurred in treated patients until now. Conclusion: Our experience describes the percutaneous PFO procedure as feasible, safe and effective with a high rate of procedural success, with an absence of significant adverse events and a high rate of complete closure. © 2013 Versita Warsaw and Springer-Verlag Berlin Heidelberg

Improving the Reprocessing Quality of Flexible Thermolabile Endoscopes: How to Learn from Mistakes

Background: Failure in the reprocessing of thermolabile flexible endoscopes has been reported as one of the most important threats to patient health. Method: A case report and observational study was conducted, from August 2014 to December 2019, in the Digestive Endoscopy Unit of a University Hospital in Italy, where two cases of Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae infections in patients undergoing endoscopic retrograde cholangio-pancreatography were observed. Following the risk/safety management practices, an epidemiological investigation was started, duodenoscopes were removed from use and the reprocessing practices reviewed. Moreover, microbiological surveillance of endoscopes was carried out according to the CDC guidelines. Results: In the first phase of sampling, 10/10 (100%) endoscopes were found to be non-compliant, of which 7 showed results for high-concern organisms (HCOs), such as KPC-K. pneumoniae, P. aeruginosa and E. coli. After implementing corrective actions, 12 out of 17 endoscopes were found to be non-compliant (70.5%), of which 8 showed results for HCOs, such as KPC-K. oxytoca and P. aeruginosa. During the last year of regular microbiological surveillance, only 23% of endoscopes (35/152) were found to be non-compliant, of which 7 showed results for HCOs, such as NDM-K. pneumoniae, P. aeruginosa and A. baumannii. The crucial issues were related to samples collected from the internal channels of duodenoscopes. Conclusion: Managing the risk associated with the reprocessing of digestive endoscopes, through risk assessment at every stage of the process, is important for the prevention of infections associated with the use of these device

Pre-Transplant Alpha-Fetoprotein &gt; 25.5 and Its Dynamic on Waitlist Are Predictors of HCC Recurrence after Liver Transplantation for Patients Meeting Milan Criteria

Background and Aim: Hepatocellular carcinoma (HCC) recurrence rates after liver transplantation (LT) range between 8 and 20%. Alpha-fetoprotein (AFP) levels at transplant can predict HCC recurrence, however a defined cut-off value is needed to better stratify patients. The aim of this study was to evaluate the rate of HCC recurrence at our centre and to identify predictors, focusing on AFP. Methods: We retrospectively analysed 236 consecutive patients that were waitlisted for HCC who all met the Milan criteria from January 2001 to December 2017 at our liver transplant centre. A total of twenty-nine patients dropped out while they were waitlisted, and 207 patients were included in the final analysis. All survival analyses included the competing-risk model. Results: The mean age was 56.8 &plusmn; 6.8 years. A total of 14% were female (n = 29/207). The median MELD (model for end-stage liver disease) at LT was 12 (9&ndash;16). The median time on the waitlist was 92 (41&ndash;170) days. The HCC recurrence rate was 16.4% (n = 34/208). The mean time to recurrence was 3.3 &plusmn; 2.8 years. The median AFP levels at transplant were higher in patients with HCC recurrence (p &lt; 0.001). At multivariate analysis, the AFP value at transplant that was greater than 25.5 ng/mL (AUC 0.69) was a strong predictor of HCC recurrence after LT [sHR 3.3 (1.6&ndash;6.81); p = 0.001]. The HCC cumulative incidence function (CIF) of recurrence at 10 years from LT was significantly higher in patients with AFP &gt; 25.5 ng/mL [34.3% vs. 11.5% (p = 0.001)]. Moreover, an increase in AFP &gt; 20.8%, was significantly associated with HCC recurrence (p = 0.034). Conclusions: In conclusion, in our retrospective study, the AFP level at transplant &gt; 25.5 ng/mL and its increase greater than 20.8% on the waitlist were strong predictors of HCC recurrence after LT in a cohort of patients that were waitlisted within the Milan criteria. However further studies are needed to validate these data