Bladder tissue characterization using probe‐based Raman spectroscopy: Evaluation of tissue heterogeneity and influence on the model prediction

Existing approaches for early‐stage bladder tumor diagnosis largely depend on invasive and time‐consuming procedures, resulting in hospitalization, bleeding, bladder perforation, infection and other health risks for the patient. The reduction of current risk factors, while maintaining or even improving the diagnostic precision, is an underlying factor in clinical instrumentation research. For example, for clinic surveillance of patients with a history of noninvasive bladder tumors real‐time tumor diagnosis can enable immediate laser‐based removal of tumors using flexible cystoscopes in the outpatient clinic. Therefore, novel diagnostic modalities are required that can provide real‐time in vivo tumor diagnosis. Raman spectroscopy provides biochemical information of tissue samples ex vivo and in vivo and without the need for complicated sample preparation and staining procedures. For the past decade there has been a rise in applications to diagnose and characterize early cancer in different organs, such as in head and neck, colon and stomach, but also different pathologies, for example, inflammation and atherosclerotic plaques. Bladder pathology has also been studied but only with little attention to aspects that can influence the diagnosis, such as tissue heterogeneity, data preprocessing and model development. The present study presents a clinical investigative study on bladder biopsies to characterize the tumor grading ex vivo, using a compact fiber probe‐based imaging Raman system, as a crucial step towards in vivo Raman endoscopy. Furthermore, this study presents an evaluation of the tissue heterogeneity of highly fluorescent bladder tissues, and the multivariate statistical analysis for discrimination between nontumor tissue, and low‐ and high‐grade tumor.     

Designing cross-linked lipase aggregates for optimum performance as biocatalysts

Cross-linked enzyme aggregates (CLEAs) are prepared by precipitation of an enzyme and then chemical cross-linking the precipitate. Three CLEAs of lipase with glutaraldehyde concentrations of 10 mM (CLEA A), 40 mM (CLEA B) and 60 mM (CLEA C) were prepared. Studies show that there is a trade-off between thermal stability vs transesterification/hydrolysis rate vs enantioselectivity. The initial rates for transesterification of β-citronellol for the uncross-linked enzyme and CLEAs A, B and C were 243, 167, 102 and 40 µmol mg^?1 h^?1, respectively. Their thermal stabilities in aqueous media, as reflected by their half-life values at 55°C, were 6, 9, 13 and 16 h, respectively. The enantioselectivity, E values (for kinetic resolution of β-citronellol by transesterification) were 19, 74, 11 and 6, respectively. These results show that CLEA C was the most thermostable; the uncross-linked enzyme was best at obtaining the highest transesterification rate; and CLEA A was best suited for the enantioselective synthesis. Scanning electron microscopy (SEM) showed that the morphology of CLEA was dependent upon the extent of cross-linking.     

Delayed egg production and a possible group effect in the blowfly Calliphora vicina

Abstract .Protein-fed Calliphora vicina , F₁ offspring of wild flies in two cages with lower and higher fly densities showed variable delay in starting oocyte vitellogenesis at ambient semi-natural temperatures in warm July–August weather in 1996 and 1997 at Durham in northern England (54°45' N). The high-density flies in 1996 showed no delay, in that the thermal sum (degree-days) experienced was 133, comparable to 18°C constant, assuming the lower threshold for egg maturation to be 5°C. Low-density cages and flies in a large outdoor cage (2 m³) in both years showed delays in production of first eggs of 34 days (thermal sum 293 degree-days) in 1996 and 32 days (396 degree-days) in 1997, and longer delays for other individuals. Delays in egg production at low densities relative to high densities seem to be a group effect of unknown mechanism.     

Prefrontal deep projection neurons enable cognitive flexibility via persistent feedback monitoring

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An X-ray spectrum estimation method from transmission measurement combined with scatter correction

PurposeConventional x-ray spectrum estimation methods from transmission measurement often lead to inaccurate results when extensive x-ray scatter is present in the measured projection. This study aims to apply the weighted L1-norm scatter correction algorithm in spectrum estimation for reducing residual differences between the estimated and true spectrum.MethodThe scatter correction algorithm is based on a simple radiographic scattering model where the intensity of scattered x-ray is directly estimated from a transmission measurement. Then, the scatter-corrected measurement is used for the spectrum estimation method that consists of deciding the weights of predefined spectra and representing the spectrum as a linear combination of the predefined spectra with the weights. The performances of the estimation method combined with scatter correction are evaluated on both simulated and experimental data.ResultsThe results show that the estimated spectra using the scatter-corrected projection nearly match the true spectra. The normalized-root-mean-square-error and the mean energy difference between the estimated spectra and corresponding true spectra are reduced from 5.8% and 1.33 keV without the scatter correction to 3.2% and 0.73 keV with the scatter correction for both simulation and experimental data, respectively.ConclusionsThe proposed method is more accurate for the acquisition of x-ray spectrum than the estimation method without scatter correction and the spectrum can be successfully estimated even the materials of the filters and their thicknesses are unknown. The proposed method has the potential to be used in several diagnostic x-ray imaging applications.     

Investigation of the magnetic susceptibility properties of fresh and fixed mouse heart,liver, skeletal muscle and brain tissue

PurposeSeveral magnetic resonance imaging (MRI) techniques exploit the difference in magnetic susceptibilities between tissues, but systematic measurements of tissue susceptibility are lacking. Furthermore, there is the question as to whether chemical fixation that is used for ex vivo MRI studies, affects the magnetic properties of the tissue. Here, we determined the magnetic susceptibility and water content of fresh and chemically fixed mouse tissue.MethodsMass susceptibility of brain, heart, liver and skeletal muscle samples were determined on a vibrating sample magnetometer at room temperature. Measurements at 50, 125, 200 and 295 K were performed to assess the temperature dependence of susceptibility. Moreover, we measured water content of fresh and fixed samples.ResultsAll samples show mass susceptibilities between −0.068 and −1.929 × 10⁻⁸ m³/kg, compared to −9.338 × 10⁻⁹ m³/kg of double distilled water. Heart tissue has a more diamagnetic susceptibility than the other tissues. Compared to fresh tissue, fixed tissue has a less diamagnetic susceptibility. Fixed tissue was not different in water content to fresh tissue and showed no consistent dependence of susceptibility with temperature, whereas fresh tissue shows a decrease to at least 125 K, indicative of a paramagnetic component.ConclusionsBiological tissues are diamagnetic in comparison to water, where the heart is more diamagnetic than the other tissues, with paramagnetic contributions. Fixation rendered tissue less diamagnetic compared to fresh tissue. Our measurements revealed differences in tissue susceptibility between VSM and QSM, inviting more research to compare susceptibility-based MRI methods with physical measurements of tissue susceptibility.     

An Ex vivo Culture System to Study Thyroid Development

The thyroid is a bilobated endocrine gland localized at the base of the neck, producing the thyroid hormones T3, T4, and calcitonin. T3 and T4 are produced by differentiated thyrocytes, organized in closed spheres called follicles, while calcitonin is synthesized by C-cells, interspersed in between the follicles and a dense network of blood capillaries. Although adult thyroid architecture and functions have been extensively described and studied, the formation of the “angio-follicular” units, the distribution of C-cells in the parenchyma and the paracrine communications between epithelial and endothelial cells is far from being understood.This method describes the sequential steps of mouse embryonic thyroid anlagen dissection and its culture on semiporous filters or on microscopy plastic slides. Within a period of four days, this culture system faithfully recapitulates in vivo thyroid development. Indeed, (i) bilobation of the organ occurs (for e12.5 explants), (ii) thyrocytes precursors organize into follicles and polarize, (iii) thyrocytes and C-cells differentiate, and (iv) endothelial cells present in the microdissected tissue proliferate, migrate into the thyroid lobes, and closely associate with the epithelial cells, as they do in vivo.Thyroid tissues can be obtained from wild type, knockout or fluorescent transgenic embryos. Moreover, explants culture can be manipulated by addition of inhibitors, blocking antibodies, growth factors, or even cells or conditioned medium. Ex vivo development can be analyzed in real-time, or at any time of the culture by immunostaining and RT-qPCR.In conclusion, thyroid explant culture combined with downstream whole-mount or on sections imaging and gene expression profiling provides a powerful system for manipulating and studying morphogenetic and differentiation events of thyroid organogenesis.