Targeting of<Emphasis Type="Italic"> lacZ</Emphasis> reporter gene expression with radioiodine-labelled phenylethyl-β-<Emphasis Type="SmallCaps">d</Emphasis>-thiogalactopyranoside

There has recently been increasing interest in the development of radioprobes that specifically target proteins transcribed from expression of reporter genes of interest. The purpose of this study was to develop a radioprobe that targets one of the most widely used reporter genes, the bacterial lacZ gene. We synthesised and purified radioiodine-labelled phenylethyl--d-thiogalactopyranoside (PETG), a competitive inhibitor specific against Escherichia coli -galactosidase. We showed that [¹²⁵I]iodo-PETG specifically binds to -galactosidase as verified by column chromatography and polyacrylamide gel electrophoresis after incubation of radiotracer with the protein. We also showed through enzyme kinetic studies that iodo-PETG retains inhibitory action against -galactosidase activity. COS-7 cells infected with a recombinant adenovirus expressing the lacZ gene had viral titre-dependent enhancements in [¹²⁵I]iodo-PETG uptake (r²=0.897; P=0.001), which reached up to 642.5%±16.7% of control levels (P<0.00001). Moreover, the level of uptake was highly correlated to luminescent measurements of -galactosidase activity (r²=0.878; P<0.0001). These results confirm that radioiodine-labelled PETG specifically targets -galactosidase and that its uptake rates faithfully reflect levels of expression of the lacZ reporter gene. Further investigations were performed in nude mice bearing human neuroblastoma tumours transferred with the lacZ gene. Compared with control tumours, lacZ-expressing tumours were slightly better visualised on [¹²³I]iodo-PETG images and had a modest increase in tumour to muscle count ratio (2.6±0.2 vs 1.9±0.1, P<0.05). The present results provide proof-of-principle for the potential of radiolabelled inhibitors as promising radiotracers to monitor lacZ gene expression levels. Future modifications to improve cell permeability should enhance in vivo contrast levels and may allow the use of radiolabelled -galactosidase inhibitors for non-invasive monitoring of lacZ gene expression.This work was presented in part at the 49th Annual Meeting of the Society of Nuclear Medicine, LA, USA, June 15–19, 2002.     

Computed tomography colonography: feasibility of computer-aided polyp detection in a "first reader" paradigm

OBJECTIVE:: To determine the feasibility of a computer-aided detection (CAD) algorithm as the "first reader" in computed tomography colonography (CTC). METHODS:: In phase 1 of a 2-part blind trial, we measured the performance of 3 radiologists reading 41 CTC studies without CAD. In phase 2, readers interpreted the same cases using a CAD list of 30 potential polyps. RESULTS:: Unassisted readers detected, on average, 63% of polyps > or =10 mm in diameter. Using CAD, the sensitivity was 74% (not statistically different). Per-patient analysis showed a trend toward increased sensitivity for polyps > or =10 mm in diameter, from 73% to 90% with CAD (not significant) without decreasing specificity. Computer-aided detection significantly decreased interobserver variability (P = 0.017). Average time to detection of the first polyp decreased significantly with CAD, whereas total reading case reading time was unchanged. CONCLUSION:: Computer-aided detection as a first reader in CTC was associated with similar per-polyp and per-patient detection sensitivity to unassisted reading. Computer-aided detection decreased interobserver variability and reduced the time required to detect the first polyp.     

Mechanical lengthening of porcine small intestine with decreased forces

Introductionshort bowel syndrome is marked by inadequate intestinal surface area to absorb nutrients. Current treatments are focused on medical management and surgical reconfiguration of the dilated intestine. We propose the use of spring-mediated distraction enterogenesis as a novel intervention to increase intestinal length. Given our previous success lengthening intestinal segments using springs with spring constant ~7 N/m that exerts 0.46 N or higher, we sought to determine the minimal force needed to lengthen porcine small intestinal segments, and to explore effects on intestine over time.MethodsJuvenile Yucatan pigs underwent laparotomy with enterotomy to introduce nitinol springs intraluminally (n = 21 springs). Bowel segments (control, spring-distracted) were retrieved on post-operative day (POD) 7 and 14, and lengths measured. Thickness of cross-sectional intestinal layers were measured using H&E, and submucosal collagen fiber orientation measured using trichrome stained sections.Resultsall pigs survived to POD7 and 14. Spring constants of at least 2 N/m exerting a minimum force of 0.10 N significantly lengthened intestinal segments (p <0.0001). The stronger the spring force, the greater the induced thickness of various intestinal layers at POD7 and 14. Collagen fiber orientation was also more disordered because of stronger springs.Conclusiona spring constant of approximately 2 N/m exerting 0.10 N and greater significantly lengthens intestinal segments and stimulates intestinal structural changes at POD7 and 14. This suggests a decreased force is capable of inducing spring-mediated distraction enterogenesis.     

Amylase level after pancreaticoduodenectomy in predicting postoperative pancreatic fistula

BackgroundIt is new clinical interest higher serum amylase level with pancreatitis after pancreaticoduodenectomy (PD) correlates with postoperative pancreatic fistula (POPF). Nevertheless, its evidence and study were scarce. We aimed to investigate correlation of serum amylase level immediate after PD and POPF occurrence.MethodsOf 163 patients who underwent PD at between January 2009 and December 2019, retrospective analysis was conducted to identify risk factors including serum amylase level immediate after PD for POPF occurrence.ResultsOverall incidence of POPF (25/163) was 15.3%. The patients occurred a POPF had significantly higher level of serum amylase on POD0 compared to in whom without a POPF (414 vs 253, p < 0.001). In univariate analysis, ASA classification, post pancreatectomy acute pancreatitis (POAP, serum amylase on POD0 >285IU/L) and Fistula Risk Grade were correlated with POPF occurrence. In multivariable analysis, Fistula risk grade and POAP were significantly associated with developing POPF.ConclusionIn patients with higher serum amylase (>285IU/L) on POD0 with higher fistula risk grade, comprehensive management to achieve mitigation of POPF is important.     

Computer-aided detection (CAD) of lung nodules in CT scans: radiologist performance and reading time with incremental CAD assistance

Objective

The diagnostic performance of radiologists using incremental CAD assistance for lung nodule detection on CT and their temporal variation in performance during CAD evaluation was assessed.

Methods

CAD was applied to 20 chest multidetector-row computed tomography (MDCT) scans containing 190 non-calcified ≥3-mm nodules. After free search, three radiologists independently evaluated a maximum of up to 50 CAD detections/patient. Multiple free-response ROC curves were generated for free search and successive CAD evaluation, by incrementally adding CAD detections one at a time to the radiologists’ performance.

Results

The sensitivity for free search was 53% (range, 44%–59%) at 1.15 false positives (FP)/patient and increased with CAD to 69% (range, 59–82%) at 1.45 FP/patient. CAD evaluation initially resulted in a sharp rise in sensitivity of 14% with a minimal increase in FP over a time period of 100 s, followed by flattening of the sensitivity increase to only 2%. This transition resulted from a greater prevalence of true positive (TP) versus FP detections at early CAD evaluation and not by a temporal change in readers’ performance. The time spent for TP (9.5 s?±?4.5 s) and false negative (FN) (8.4 s?±?6.7 s) detections was similar; FP decisions took two- to three-times longer (14.4 s?±?8.7 s) than true negative (TN) decisions (4.7 s?±?1.3 s).

Conclusions

When CAD output is ordered by CAD score, an initial period of rapid performance improvement slows significantly over time because of non-uniformity in the distribution of TP CAD output and not to a changing reader performance over time.     

Dose-responsiveness of electrophysiologic change in a new model of acute carpal tunnel syndrome

This study was done to determine the dose-responsiveness during the development of acute pressure-induced median neuropathy in rabbits and to develop a new animal model of acute carpal tunnel syndrome. Twenty-three rabbits were used. Carpal tunnel syndrome was induced by infusing a controlled saline solution into the carpal tunnel of rabbits using a 21-gauge needle under general anesthesia to elevate the carpal tunnel pressure. The changes in the amplitude and latency of the compound muscle action potential obtained by abductor pollicis recording were observed after a complete conduction block, and after the release of pressure at various pressure levels. Pressures greater than 30 mm Hg applied to the carpal tunnel caused a remarkable electrophysiologic change. A higher pressure level resulted in a shorter time required for a complete conduction block and also for recovery after the release of pressure. Complete conduction block occurred between 40 and 50 minutes at a pressure of 100 mm Hg. The degree of recovery after a conduction block was related inversely to the pressure-time integrals. Our animal model reflects the pathophysiology of acute carpal tunnel syndrome and shows a regular dose-responsiveness during the development of acute pressure-induced neuropathy.     

Prevention of graft rejection following bone marrow transplantation

Bone marrow transplantation from an HLA-identical sibling is increasingly used in the treatment of severe aplastic anemia. One major problem with this approach is graft rejection that occurs in 25%-60% of patients conditioned for transplantation with cyclophosphamide. At most transplant centers it has been difficult to accurately identify patients at high risk for graft rejection. We studied a conditioning regimen of cyclophosphamide (200 mg per kg) and low-dose total body irradiation (3 Gy; equivalent to 300 rad) in 23 consecutive unselected patients with aplastic anemia followed for a minimum of 6 mo. There was only one episode of graft rejection. Graft-versus-host disease and interstitial pneumonitis were not increased by the more intensive conditioning regimen. Actuarial survival was 61% at 1 yr and 49% at 2.5 yr. Cyclophosphamide and low-dose total body irradiation is an effective conditioning regimen in patients with aplastic anemia. It may be particularly useful when accurate predictive tests of graft rejection are not available as is the case in most transplant centers.     

COVID-19 and Risk Factors of Anxiety and Depression in South Korea

ObjectiveThe aims of this study were to explore the prevalence of and identify predictors of anxiety and depression related to coronavirus disease of 2019 (COVID-19) in South Korea. MethodsThe analysis is based on a quota survey design and a sampling frame that permitted recruitment of a national sample of 1,014 individuals between March 17–31, 2020. Several standardized measurements were used, including GAD-7, PHQ-9, COVID-19 related fear, restrictions in deaily life, as well as sociodemographic information and physical and psychosocial needs during the pandemic. Multiple logistic regression was conducted to analyze the influence of sociodemographic factors, fear, and physical/psychosocial needs on anxiety and depression. ResultsSignificant numbers of the respondents were identifiable anxiety (19.0%) and depression group (17.5%), respectively. This indicates that the depression and anxiety prevalence rate after the COVID-19 is substantially high compared to the depression rate of 2.6% in 2020 and 2.8% in 2018 both reported in the Korea Community Health Survey and the anxiety rate of 5.7% reported in 2016 Survey of Mental Disorders in Korea. Multiple logistic regression results showed age, COVID-19 related fear, and the level of restrictions in daily as significant factors in understanding and predicting the anxiety group. Likfewise, the COVID-19 related fear, restrictions in daily life, and need for economic support were important predictors in predicting the depression group. ConclusionFindings on predictors for greater vulnerability to anxiety and depression has important implications for public mental health in the context of the COVID-19 pandemic.     

The Impact of Emotional Exhaustion on Psychological Factors in Workers with Secondary Traumatic Experiences: A Multi-Group Path Analysis

Objective The objective of the present study was to explore causal pathways to understand how second traumatic experiences could affect the development of emotional exhaustion and psychiatric problems. Methods A total of 582 workers who had jobs vulnerable to secondary traumatic experiences were enrolled for this study. Emotional exhaustion, secondary trauma, resilience, perceived stress, depression, anxiety, and sleep problems were evaluated. A model with pathways from secondary traumatic experience score to depression and anxiety was proposed. The participants were divided into three groups according to the resilience: the low, middle and high resilience group. Results Resilience was a meaningful moderator between secondary traumatic experiences and psychiatric problems. In the path model, the secondary trauma and perceived stress directly and indirectly predicted perceived stress, emotional exhaustion, depression, anxiety, and sleep problems in all three groups. Direct effects of perceived stress on depression and anxiety were the largest in the low resilience group. However, direct effects of secondary trauma on perceived stress and emotional exhaustion were the largest in the high resilience group. Conclusion Understanding the needs of focusing for distinct psychological factors offers a valuable direction for the development of intervention programs to prevent emotional exhaustion among workers with secondary traumatic experiences.