Comparison of manual and automated digital image analysis systems for quantification of cellular protein expression.

OBJECTIVE: Quantifying protein expression in immunohistochemically stained histological slides is an important tool for oncologic research. The use of computer-aided evaluation of IHC-stained slides significantly contributes to objectify measurements. Manual digital image analysis (mDIA) requires a user-dependent annotation of the region of interest (ROI). Others have built-in machine learning algorithms with automated digital image analysis (aDIA) and can detect the ROIs automatically. We aimed to investigate the agreement between the results obtained by aDIA and those derived from mDIA systems. METHODS: We quantified chromogenic intensity (CI) and calculated the positive index (PI) in cohorts of tissue microarrays (TMA) using mDIA and aDIA. To consider the different distributions of staining within cellular sub-compartments and different tumor architecture our study encompassed nuclear and cytoplasmatic stainings in adenocarcinomas and squamous cell carcinomas. RESULTS: Within all cohorts, we were able to show a high correlation between mDIA and aDIA for the CI (p<0.001) along with high agreement for the PI. Moreover, we were able to show that the cell detections of the programs were comparable as well and both proved to be reliable when compared to manual counting. CONCLUSION: mDIA and aDIA show a high correlation in acquired IHC data. Both proved to be suitable to stratify patients for evaluation with clinical data. As both produce the same level of information, aDIA might be preferable as it is time-saving, can easily be reproduced, and enables regular and efficient output in large studies in a reasonable time period.

Retrospective analysis of time-related three-dimensional iliac bone graft resorption following sinus lift and vertical augmentation in the maxilla.

The atrophic maxilla frequently requires bone grafting using an onlay graft (OG) or sinus lifting (SL) before implant rehabilitation. The resorption of bone grafts is influenced by the time until implantation, quality of donor bone, and grafting technique. The aim of this study was to investigate the impact of both grafting techniques on the time-related resorption of autologous iliac bone graft. Forty-three patients underwent either onlay grafting or a sinus lift at 73 sites in the maxilla. Graft height was measured by cone beam computed tomography after augmentation and during follow-up for up to 12 months prior to implant insertion. The effect of time and technique on graft resorption was evaluated retrospectively. The reduction in bone graft height was greater for OG than SL over the investigated time intervals (OG = 51%, SL = 28%; P = 0.002). Each technique followed a specific course of resorption, which was independent of the initial graft height and could be calculated by a non-linear regression model. Iliac bone graft undergoes rapid resorption when used as an OG prior to implant insertion. For SL, this resorption is reasonably lower. This is especially crucial to determine the optimal time for implant insertion after graft healing to improve implant survival.

Impacts of CO2 Enrichment on Productivity and Light Requirements of Eelgrass.

Seagrasses, although well adapted for submerged existence, are CO2-limited and photosynthetically inefficient in seawater. This leads to high light requirements for growth and survival and makes seagrasses vulnerable to light limitation. We explored the long-term impact of increased CO2 availability on light requirements, productivity, and C allocation in eelgrass (Zostera marina L.). Enrichment of seawater CO2 increased photosynthesis 3-fold, but had no long-term impact on respiration. By tripling the rate of light-saturated photosynthesis, CO2 enrichment reduced the daily period of irradiance-saturated photosynthesis (Hsat) that is required for the maintenance of positive whole-plant C balance from 7 to 2.7 h, allowing plants maintained under 4 h of Hsat to perform like plants growing in unenriched seawater with 12 h of Hsat. Eelgrass grown under 4 h of Hsat without added CO2 consumed internal C reserves as photosynthesis rates and chlorophyll levels dropped. Growth ceased after 30 d. Leaf photosynthesis, respiration, chlorophyll, and sucrose-phosphate synthase activity of CO2-enriched plants showed no acclimation to prolonged enrichment. Thus, the CO2-stimulated improvement in photosynthesis reduced light requirements in the long term, suggesting that globally increasing CO2 may enhance seagrass survival in eutrophic coastal waters, where populations have been devastated by algal proliferation and reduced water-column light transparency.

Carbon Partitioning in Eelgrass (Regulation by Photosynthesis and the Response to Daily Light-Dark Cycles).

Diel variations in rates of C export, sucrose-phosphate synthase (SPS) and sucrose synthase (SS) activity, and C reserves were investigated in Zostera marina L. (eelgrass) to elucidate the environmental regulation of sucrose formation and partitioning in this ecologically important species. Rates of C flux and SPS activity increased with leaf age, consistent with the ontogenic transition from sink to source status. Rates of C export and photosynthesis were low but quantitatively consistent with those of many terrestrial plant species. The Vmax activity of SPS approached that of maize, but substrate-limited rates were 20 to 25% of Vmax, indicating a large pool of inactive SPS. SPS was unresponsive to the day/night transition or to a 3-fold increase in photosynthesis generated by high [CO2] and showed little sensitivity to inorganic phosphate. Consequently, regulation of eelgrass SPS appeared similar to starch- rather than to sugar-accumulating species even though eelgrass accumulates sucrose. Leaf [sucrose] was constant and high throughout the diel cycle, which may contribute to the down-regulation of SPS. Root sucrose synthase activity was high but showed no response to nocturnal anoxia. Root [sucrose] also showed no diel cycle. The temporal stability of [sucrose] confers an ability for eelgrass to buffer the effects of prolonged light limitation that may be key to its survival and ecological success in environments subject to periods of extreme light limitation and chaotic daily variation in light availability.

Quality control of storage phosphor digital radiography systems.

Quality control (QC) of storage phosphor devices is important in assuring that the image information entered into an Image management and communication (IMAC) system is sufficient for diagnosis. QC of storage phosphor digital radiography systems is complex because of the self-corrective nature of the image-processing software used in these machines. Currently, one must produce hard copy to perform adequate QC. Inspection of images with reject analysis and inspection of cassettes and imaging plates has helped us in our QC program. For those QC tests using control limits, the appropriate settings for these limits are unknown. Starting approximations are given. Recommended tests are described.

Digital radiography of the musculoskeletal system: the optimal image.

Digital radiography (DR) is replacing screen-film (SF) radiography for musculoskeletal examinations in our institution. After an iterative process of image quality improvement, our DR images are now preferred to SF images, and we have detailed our current optimized settings for the Fuji 9000 (Fuji Medical Systems, Tokyo, Japan). DR offers the advantages of improved contrast resolution, adjustable image contrast, the ability to reprocess the image, and the ease of transferring the image to an image management and communication system (IMAC).

Detection of aortic dissection by transoesophageal echocardiography.

The diagnostic value of a combination of transoesophageal and transthoracic echocardiography was evaluated in 21 patients with dissection of the aorta. The results were compared with those of computed tomography, aortography, and with findings at operation or necropsy or both. Transthoracic echocardiography identified three of the four patients with type I dissection, two of the five patients with type II dissection, and one of the 12 patients with type III dissection. When transoesophageal echocardiography was used as well the degree of aortic dissection was identified correctly in all 21 patients. In one patient with type I and in eight patients with type III dissection spontaneous echocardiographic contrast with a mural thrombus within the false lumen could be detected. Computed tomography was unable to demonstrate an intimal flap in one of two patients studied with type I dissection, in two of three patients with type II dissection, and in one of nine patients with type III dissection. Aortography was negative in one of two patients studied with type I dissection, two of four patients with type II dissection, and in one of eight patients with type II dissection. The whole thoracic aorta can be imaged by a combination of transthoracic and transoesophageal echocardiography. The addition of transoesophageal echocardiography to transthoracic echocardiography improves the recognition of aortic dissection. Furthermore, this examination can be performed at the bedside and the findings can be used as a basis for treatment.

Aortic dissection detected by transoesophageal echocardiography.

The diagnostic value of transoesophageal echocardiography was evaluated in 24 patients with aortic dissection and compared to transthoracic two-dimensional echocardiography, computer tomography, aortography, surgery and autopsy. Using transoesophageal echocardiography we found in 5 patients a type I dissection, in 5 patients a type II and in 14 patients a type III dissection. Transthoracic two-dimensional echocardiography was positive in 3/5 type I, 2/5 in type II and 2/14 in type III dissections. Computer tomography was unable to demonstrate an intimal flap in 1/2 patients with type I, 2/3 type II and 1/11 type III dissections. Aortography was negative in 1/4 type I, 3/5 type II and 3/12 patients with type III dissection. Additional information concerning thrombus formation, localisation of the entry tear, differentiation between true and false lumen, flow dynamics within the true and false lumen as well as accompanying aortic regurgitation may be obtained by transoesophageal echocardiography.