Enumeration of leukocyte infiltration in solid tumors by confocal laser scanning microscopy

BACKGROUND: Leukocytes commonly infiltrate solid tumors, and have been implicated in the mechanism of spontaneous regression in some cancers. Conventional techniques for the quantitative estimation of leukocyte infiltrates in tumors rely on light microscopy of immunostained thin tissue sections, in which an arbitrary assessment (based on low, medium or high levels of infiltration) of antigen density is made by the pathologist. These estimates are relatively subjective and often require the opinion of a second pathologist. In addition, since thin tissue sections are cut, no data regarding the three-dimensional distribution of antigen can be obtained. RESULTS: To overcome these problems, we have designed a method to enumerate leukocyte infiltration into tumors, using confocal laser scanning microscopy of fluorescently immunostained leukocytes in thick tissue sections. Using image analysis software, a threshold was applied to eliminate unstained tissue and residual noise. The total antigen volume in the scanned tissue was calculated and divided by the mean cell volume (calculated by "seeding" ten individual cells) to obtain the cell count. Using this method, we compared the calculated leukocyte counts with those obtained manually by ten laboratory personnel. There was no significant difference (P > 0.05) between the cell counts obtained by either method. We then compared leukocyte infiltration into seven tumors and matched non-malignant tissue obtained from the periphery of the resected tissue. There was a significant increase in the infiltration of all leukocyte subsets into the tumors compared to minimal numbers in the non-malignant tissue. CONCLUSION: From these results we conclude that this method may be of considerable use for the enumeration of cells in tissues. Furthermore, since it can be performed by laboratory technical staff, less time input is required by the pathologist in assessing the degree of leukocyte infiltration into tumors

Crk and CrkL adaptor proteins: networks for physiological and pathological signaling

The Crk adaptor proteins (Crk and CrkL) constitute an integral part of a network of essential signal transduction pathways in humans and other organisms that act as major convergence points in tyrosine kinase signaling. Crk proteins integrate signals from a wide variety of sources, including growth factors, extracellular matrix molecules, bacterial pathogens, and apoptotic cells. Mounting evidence indicates that dysregulation of Crk proteins is associated with human diseases, including cancer and susceptibility to pathogen infections. Recent structural work has identified new and unusual insights into the regulation of Crk proteins, providing a rationale for how Crk can sense diverse signals and produce a myriad of biological responses

Oxidation studies of Fe10CrAl-RE alloys exposed to Pb at 550 degrees C for 10,000 h

Five experimental FeCrAl-RE alloys have been exposed up to 10,000 h in stagnant liquid Pb at 550 C. The test matrix consisted of three 10 wt.% Cr alloys, with an Al content ranging from 4 to 8 wt.% (10Cr-4Al, 10Cr-6Al and 10Cr-8Al), one alloy without additions of reactive elements (RE) (10Cr-6Al), and one reference alloy with 21 wt.% Cr and 5 wt.% Al (21Cr-5Al). The evaluation showed a clear difference in oxidation properties, and it was possible to divide the alloys into two distinct groups. A critical Al concentration in the interval of 4-6 wt.% at the given RE content was required to form a thin protective oxide. However, the absence of RE addition in one of the two 10Cr-6Al alloys resulted in a significant reduction in oxidation resistance, comparable with 10Cr-4Al. None of the alloys were severely corroded, however Pb penetrated to a relatively large extent into the porous oxide of the low performing alloys. A 100 nm thick oxide scale, partly consisting of alumina (Al2O3), was observed for the high performing 10Cr-6Al alloy. The Fe10CrAl-RE alloys showed overall very good corrosion resistance and are hence a promising new alloy category for liquid Pb applications. (C) 2013 Elsevier B.V. All rights reserved

Oxidation studies of Fe10CrAl-RE alloys exposed to Pb at 550 degrees C for 10,000 h

Five experimental FeCrAl-RE alloys have been exposed up to 10,000 h in stagnant liquid Pb at 550 C. The test matrix consisted of three 10 wt.% Cr alloys, with an Al content ranging from 4 to 8 wt.% (10Cr-4Al, 10Cr-6Al and 10Cr-8Al), one alloy without additions of reactive elements (RE) (10Cr-6Al), and one reference alloy with 21 wt.% Cr and 5 wt.% Al (21Cr-5Al). The evaluation showed a clear difference in oxidation properties, and it was possible to divide the alloys into two distinct groups. A critical Al concentration in the interval of 4-6 wt.% at the given RE content was required to form a thin protective oxide. However, the absence of RE addition in one of the two 10Cr-6Al alloys resulted in a significant reduction in oxidation resistance, comparable with 10Cr-4Al. None of the alloys were severely corroded, however Pb penetrated to a relatively large extent into the porous oxide of the low performing alloys. A 100 nm thick oxide scale, partly consisting of alumina (Al2O3), was observed for the high performing 10Cr-6Al alloy. The Fe10CrAl-RE alloys showed overall very good corrosion resistance and are hence a promising new alloy category for liquid Pb applications. (C) 2013 Elsevier B.V. All rights reserved

Effects of alloying concepts on ferrite morphology and toughness of lean duplex stainless steel weld metals

Alloying concepts for lean duplex weld metals have been explored. Focus was on compositions with 21.5-24 % Cr with different levels of Ni and Mn in combination with N producing a minimum PREN value of 26. Microstructures and properties of weld metals produced with experimental covered electrodes or metal-cored wires were evaluated. The electron backscattered diffraction (EBSD) technique combined with scanning electron microscopy (SEM) was used to explore relations between weld metal morphology and crystallographic orientation relationships between ferrite and austenite. A duplex microstructure with Widmanstatten type ferrite characteristic of a ferritic solidification was found throughout most weld metals. Regions with a morphology suggesting a mixed ferritic-austenitic solidification occurred particularly for higher Ni- and N-contents. EBSD studies showed that most of the austenite formed following a ferritic solidification was near either the Nishiyama-Wasserman (NW) or Kurjdumov-Sachs (KS) relationships with adjacent ferrite. More phase boundaries in the mixed mode solidification regions had a random orientation relationship. Strength, ductility and PREN requirements were readily matched but the combined Mn, Ni and N alloying levels and the ferrite morphology affected toughness significantly. Both a fully ferritic solidification and a sufficient Ni-content were necessary to produce acceptable and reproducible impact toughness at room and sub-zero temperatures. It was concluded that a composition of 23-24 % Cr, 7-8 % Ni and 0.12-0.16 % N was well suited to fulfil requirements

The Influence of Heat Treatment on the Microstructure and Machinability of a Prehardened Mold Steel

© 2015, The Minerals, Metals & Materials Society and ASM International. The machinability performance of a modified AISI P20 steel, heat treated to have the same hardness but three different microstructures, lower bainite, tempered martensite, and primary spheroidized carbides in a tempered martensite matrix, was studied. The microstructures were characterized using light optical and scanning electron microscopy and X-ray diffraction, and mechanical properties were compared by means of tensile and Charpy V-notch impact tests. The influence of microstructure and the resultant mechanical properties on machinability was studied in the context of single tooth end milling operation. The results showed that the material containing primary spheroidized carbides exhibited a superior machinability at the expense of a marginal loss of tensile strength and impact toughness, with comparable yield strength to that of the material containing tempered martensite. By contrast, the material with bainitic microstructure showed the lowest yield strength and the poorest machinability performance while having the highest uniform elongation.Peer Reviewe

A Novel Fluorescence Imaging Technique Combining Deconvolution Microscopy And Spectral Analysis For Quantitative Detection Of Opportunistic Pathogens

A novel fluorescence imaging technique based on deconvolution microscopy and spectral analysis is presented here as an alternative to confocal laser scanning microscopy. It allowed rapid, specific and simultaneous identification of five major opportunistic pathogens, relevant for public health, in suspension and provided quantitative results. © 2006 Elsevier B.V. All rights reserved