Diagnostic Efficacy of PET/CT Plus Brain MR Imaging for Detection of Extrathoracic Metastases in Patients with Lung Adenocarcinoma

We aimed to evaluate prospectively the efficacy of positron emission tomography (PET)/computed tomography (CT) plus brain magnetic resonance imaging (MRI) for detecting extrathoracic metastases in lung adenocarcinoma. Metastatic evaluations were feasible for 442 consecutive patients (M:F=238:204; mean age, 54 yr) with a lung adenocarcinoma who underwent PET/CT (CT, without IV contrast medium injection) plus contrast-enhanced brain MRI. The presence of metastases in the brain was evaluated by assessing brain MRI or PET/CT, and in other organs by PET/CT. Diagnostic efficacies for metastasis detection with PET/CT plus brain MRI and with PET/CT only were calculated on a per-patient basis and compared from each other. Of 442 patients, 88 (20%, including 50 [11.3%] with brain metastasis) had metastasis. Regarding sensitivity of overall extrathoracic metastasis detection, a significant difference was found between PET/CT and PET/CT plus brain MRI (68% vs. 84%; P=0.03). As for brain metastasis detection sensitivity, brain MRI was significantly higher than PET/CT (88% vs. 24%; P<0.001). By adding MRI to PET/CT, brain metastases were detected in additional 32 (7% of 442 patients) patients. In lung adenocarcinoma patients, significant increase in sensitivity can be achieved for detecting extrathoracic metastases by adding dedicated brain MRI to PET/CT and thus enhancing brain metastasis detection

Establishment of Functioning Human Corneal Endothelial Cell Line with High Growth Potential

Hexagonal-shaped human corneal endothelial cells (HCEC) form a monolayer by adhering tightly through their intercellular adhesion molecules. Located at the posterior corneal surface, they maintain corneal translucency by dehydrating the corneal stroma, mainly through the Na+- and K+-dependent ATPase (Na+/K+-ATPase). Because HCEC proliferative activity is low in vivo, once HCEC are damaged and their numbers decrease, the cornea begins to show opacity due to overhydration, resulting in loss of vision. HCEC cell cycle arrest occurs at the G1 phase and is partly regulated by cyclin-dependent kinase inhibitors (CKIs) in the Rb pathway (p16-CDK4/CyclinD1-pRb). In this study, we tried to activate proliferation of HCEC by inhibiting CKIs. Retroviral transduction was used to generate two new HCEC lines: transduced human corneal endothelial cell by human papillomavirus type E6/E7 (THCEC (E6/E7)) and transduced human corneal endothelial cell by Cdk4R24C/CyclinD1 (THCEH (Cyclin)). Reverse transcriptase polymerase chain reaction analysis of gene expression revealed little difference between THCEC (E6/E7), THCEH (Cyclin) and non-transduced HCEC, but cell cycle-related genes were up-regulated in THCEC (E6/E7) and THCEH (Cyclin). THCEH (Cyclin) expressed intercellular molecules including ZO-1 and N-cadherin and showed similar Na+/K+-ATPase pump function to HCEC, which was not demonstrated in THCEC (E6/E7). This study shows that HCEC cell cycle activation can be achieved by inhibiting CKIs even while maintaining critical pump function and morphology

Minimal information for studies of extracellular vesicles (MISEV2023): From basic to advanced approaches

Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its 'Minimal Information for Studies of Extracellular Vesicles', which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly

Macular Bruch's membrane defect and dome-shaped macula in high myopia.

To examine an association between macular Bruch's membrane defects (MBMD) and a dome-shaped appearance of the macula (DSM).Retrospective, observational case series study.The study included highly myopic individuals who were consecutively examined between May 2014 and December 2015. The patients underwent swept-source optical coherence tomography (OCT) for visualization of DSM and MBMDs defined as Bruch´s membrane defects located at a distance of maximal 1500 μm from the foveola.Out of 1983 highly myopic eyes (1057 patients), 166 eyes (8.4%; 95% confidence interval (CI):7.2%,9.6%)) showed a DSM and 534 eyes showed a MBMD. In multivariate binary regression analysis, higher prevalence of DSM was associated with a higher prevalence of a MBMD (P<0.001; OR: 1.96; 95%CI: 1.40, 2.75) after adjusting for longer axial length (P<0.001; odds ratio (OR): 1.27; 95%CI: 1.16, 1.38). In eyes with a DSM partially surrounded by a MBMD, the retina, retinal pigment epithelium (RPE) and choroid appeared relatively unchanged in the central region with Bruch´s membrane (BM) preserved. In the ring-like BM-free region surrounding the central prominent island of the DSM, the RPE, the outer and middle retinal layers, the choriocapillaris and the middle-sized choroidal vessel layer were absent. In association with a DSM, three MBMD types were differentiated: MBMDs in patchy chorioretinal atrophy, MBMDs in choroidal neovascularization-related macular atrophy, and MBMDs as temporally extending large parapapillary gamma zone.Presence of a DSM was significantly associated with the presence of MBMDs. The morphology of the DSM in association with MBMDs may be associated with a focal relaxation of the posterior sclera, no longer pushed outward by an expanding BM but allowed to partially bulge inward, leading to the formation of a DSM

Dome-shaped macula with protrusion of inner part of sclera

Purpose: To report our findings in a patient with a two layered dome shaped macula (DSM) in which only the inner layer of the sclera protruded anteriorly. Observations: An 84-year-old woman with high myopia had a DSM in both eyes. The optical coherence tomographic (OCT) image of the left eye showed a uniform thickening of the foveal sclera, but the DSM of the right eye was split into an inner and outer layer by intrascleral blood vessels running between the two layers. OCT showed that only the inner layer of the sclera protruded anteriorly while the outer layer remained in its normal position. Conclusions and importance: The two layered DSM suggests that the etiology of DSMs may be more complex

Posterior staphylomas and scleral curvature in highly myopic children and adolescents investigated by ultra-widefield optical coherence tomography.

PurposeTo determine the early signs of posterior staphylomas in highly myopic eyes of younger subjects by swept-source ultra-widefield optical coherence tomography (WF-OCT).MethodsThis was an observational case series study. Highly myopic subjects younger than 20 years old who were examined consecutively by prototype WF-OCT were studied. High myopia was defined according to the Ministry of Health and Welfare, Japan classification. A posterior displacement of the sclera and two OCT features indicating the staphyloma edges were used as markers of a staphyloma.ResultsFifty-five eyes of 30 patients with the mean age of 12.3 years, and the mean axial length of 27.9 mm were studied. Seven of the 55 eyes (12.7%) had a posterior displacement of the sclera and were diagnosed as having a staphyloma. Among the two OCT features of the staphyloma edges, a gradual thinning of the choroid toward the staphyloma edge and gradual re-thickening of choroid from the staphyloma edge toward the posterior pole were found in these 7 eyes. However, the other feature of an inward protrusion of the sclera at the staphyloma edge, was obvious in only 2 eyes. The subfoveal choroid and choroid nasal to the optic disc were significantly thinner in eyes with a staphyloma than those without it.ConclusionsThe changes of the choroidal thickness toward the staphyloma edge with the posterior displacement of the sclera were considered an early sign which precedes an inward protrusion of sclera at the staphyloma edge

Tumorigenesis assay of cell lines in BALB/C nude mice.

<p>Tumorigenesis assay of cell lines in BALB/C nude mice.</p

Transmission electron microscopy of cell line intercellular junctions.

<p>The junctional complex was detected at the intercellular junction in THCEC (Cyclin) and HCEC. No component of the intercellular junction was found in THCEC (E6/E7), in which cells grew in multilayers without being inhibited by cellular contact (scale bar = 200 nm).</p

The pump function of cell lines.

<p>Short-circuit currents representing Na⁺/K+-ATPase activity from corneal cell monolayers on the insert well area of 4.67 cm² were calculated before and after addition of the Na⁺/K+-ATPase inhibitor ouabain. (A) Representative tracings of short-circuit current (<i>µ</i>A/well) obtained with cell monolayers of THCEC (Cyclin) (upper panel), HCEC (middle panel) and THCEC (E6/E7) (lower panel). THCEC (Cyclin) possessed equal transport activity to HCEC, whereas no pump function was detected in THCEC (E6/E7). (B) Time-course changes in the average short circuit current of cultured monolayers of cell lines at 1, 5, 10 and 20 min. Data shown are for (▴) THCEC (Cyclin) at PD8, (♦) THCEC (Cyclin) at PD 21, (•) HCEC and (▪) THCEC (E6/E7); all data are expressed as mean±SD of four replicate experiments of each cell line.</p

Evaluation of proliferative capacity.

<p>(A) Immunohistochemistry of Ki-67 in three cell lines. Positive staining of Ki-67, located in the nucleus, was obviously identified in THCEC (Cyclin) and THCEC (E6/E7), but rarely detected in HCEC. (B) Real-time PCR of downstream genes of cyclinD1 associated with proliferation. Gene expression levels of both CDC2 and PCAN were clearly higher than that of HCEC. The gene expression was much more activated in THCEC (Cyclin) in which the expression of E2F, an upstream transcriptional factor of two genes, was constitutively activated by transduced mutant Cdk4 and CyclinD1.</p