Post-mortem computed tomography is a useful tool for determining the pulmonary ventilation status in newborns

<jats:title>Abstract</jats:title><jats:sec> <jats:title>Introduction</jats:title> <jats:p>Lung ventilation is a standard sign of life in newborns. Post-mortem computed tomography (PMCT) is highly sensitive to the presence of gas in the body including the lungs. Current standard examinations to determine the pulmonary ventilation status in newborns are the flotation test and histology. The purpose of this study was to compare the accuracy of PMCT with the flotation test for determining the lung ventilation status with histological control as reference standard. A cut-off value as CT number in Hounsfield Units (HU) determining lung ventilation of newborns in PMCT should be established.</jats:p> </jats:sec><jats:sec> <jats:title>Methods</jats:title> <jats:p>A total of 38 infant lungs were examined of which 21 lungs were from infants deceased shortly after live birth (control group) and 17 lungs belonged to infants where live birth was in question (study group). All lungs were examined using PMCT, flotation test, and histological examination.</jats:p> </jats:sec><jats:sec> <jats:title>Results</jats:title> <jats:p>The control group showed an overall mean attenuation ± standard deviation of −219 HU ± 135; the study group of 45 ± 15 HU in histologically nonventilated lungs versus −192 ± 207 HU; (<jats:italic>p</jats:italic> < 0.001) in ventilated lungs. The best cut-off value for optimal discrimination of ventilated and nonventilated lungs in newborns was −35 mean HU.</jats:p> </jats:sec><jats:sec> <jats:title>Conclusion</jats:title> <jats:p>PMCT is equally well suited to determine lung ventilation as the flotation test. It provides additional information regarding pulmonary infiltration, degree of putrefaction, or signs of trauma (fractures, pneumothorax). Histology remains mandatory in ambiguous cases.</jats:p> </jats:sec&gt

Increased Circulating T Cell Reactivity to GM1 Ganglioside in Patients with Guillain-Barre Syndrome

This study was performed to determine whether increased ganglioside-specific T cell reactivity can be detected in the peripheral blood of patients with Guillain-Barre syndrome (GBS) and chronic inflammatory demyelinating polyradiculoneuropathy (CIDP). T cell responsiveness to the gangliosides GM1, GM3, GD1a, GD1b, GD3, GT1b, GQ1b and sulphatide was assessed in peripheral blood mononuclear cells from untreated GBS patients (57), CIDP patients (43), patients with other peripheral neuropathies (55) and healthy control subjects (74) in a standard 6-day proliferation assay. Increased T cell reactivity to GM1 occurred in GBS patients compared to healthy controls and patients with other neuropathies. There was increased reactivity to GM3 in GBS patients compared to patients with other neuropathies but not compared to healthy controls. The frequencies of increased T cell reactivity to GM1 and GM3 in CIDP patients were intermediate between those of GBS patients and controls. We suggest that T cell reactivity to gangliosides might play a contributory role in the pathogenesis of GBS and perhaps CIDP

Automated Steel Cleanliness Analysis Tool (ASCAT)

The objective of this study was to develop the Automated Steel Cleanliness Analysis Tool (ASCATTM) to permit steelmakers to evaluate the quality of the steel through the analysis of individual inclusions. By characterizing individual inclusions, determinations can be made as to the cleanliness of the steel. Understanding the complicating effects of inclusions in the steelmaking process and on the resulting properties of steel allows the steel producer to increase throughput, better control the process, reduce remelts, and improve the quality of the product. The ASCAT (Figure 1) is a steel-smart inclusion analysis tool developed around a customized next-generation computer controlled scanning electron microscopy (NG-CCSEM) hardware platform that permits acquisition of inclusion size and composition data at a rate never before possible in SEM-based instruments. With built-in customized ''intelligent'' software, the inclusion data is automatically sorted into clusters representing different inclusion types to define the characteristics of a particular heat (Figure 2). The ASCAT represents an innovative new tool for the collection of statistically meaningful data on inclusions, and provides a means of understanding the complicated effects of inclusions in the steel making process and on the resulting properties of steel. Research conducted by RJLG with AISI (American Iron and Steel Institute) and SMA (Steel Manufactures of America) members indicates that the ASCAT has application in high-grade bar, sheet, plate, tin products, pipes, SBQ, tire cord, welding rod, and specialty steels and alloys where control of inclusions, whether natural or engineered, are crucial to their specification for a given end-use. Example applications include castability of calcium treated steel; interstitial free (IF) degasser grade slag conditioning practice; tundish clogging and erosion minimization; degasser circulation and optimization; quality assessment/steel cleanliness; slab, billet or bloom disposition; and alloy development. Additional benefits of ASCAT include the identification of inclusions that tend to clog nozzles or interact with refractory materials. Several papers outlining the benefits of the ASCAT have been presented and published in the literature. The paper entitled ''Inclusion Analysis to Predict Casting Behavior'' was awarded the American Iron and Steel Institute (AISI) Medal in 2004 for special merit and importance to the steel industry. The ASCAT represents a quantum leap in inclusion analysis and will allow steel producers to evaluate the quality of steel and implement appropriate process improvements. In terms of performance, the ASCAT (1) allows for accurate classification of inclusions by chemistry and morphological parameters, (2) can characterize hundreds of inclusions within minutes, (3) is easy to use (does not require experts), (4) is robust, and (5) has excellent image quality for conventional SEM investigations (e.g., the ASCAT can be utilized as a dual use instrument). In summary, the ASCAT will significantly advance the tools of the industry and addresses an urgent and broadly recognized need of the steel industry. Commercialization of the ASCAT will focus on (1) a sales strategy that leverages our Industry Partners; (2) use of ''technical selling'' through papers and seminars; (3) leveraging RJ Lee Group's consulting services, and packaging of the product with a extensive consulting and training program; (4) partnering with established SEM distributors; (5) establishing relationships with professional organizations associated with the steel industry; and (6) an individualized plant by plant direct sales program

Expression of Abelson Interactor 1 (Abi1) Correlates with Inflammation, KRAS Mutation and Adenomatous Change during Colonic Carcinogenesis

<div><h3>Background</h3><p>Abelson interactor 1 (Abi1) is an important regulator of actin dynamics during cytoskeletal reorganization. In this study, our aim was to investigate the expression of Abi1 in colonic mucosa with and without inflammation, colonic polyps, colorectal carcinomas (CRC) and metastases as well as in CRC cell lines with respect to BRAF/KRAS mutation status and to find out whether introduction of KRAS mutation or stimulation with TNFalpha enhances Abi1 protein expression in CRC cells.</p> <h3>Methodology/Principal Findings</h3><p>We immunohistochemically analyzed Abi1 protein expression in 126 tissue specimens from 95 patients and in 5 colorectal carcinoma cell lines with different mutation status by western immunoblotting. We found that Abi1 expression correlated positively with KRAS, but not BRAF mutation status in the examined tissue samples. Furthermore, Abi1 is overexpressed in inflammatory mucosa, sessile serrated polyps and adenomas, tubular adenomas, invasive CRC and CRC metastasis when compared to healthy mucosa and BRAF-mutated as well as KRAS wild-type hyperplastic polyps. Abi1 expression in carcinoma was independent of microsatellite stability of the tumor. Abi1 protein expression correlated with KRAS mutation in the analyzed CRC cell lines, and upregulation of Abi1 could be induced by TNFalpha treatment as well as transfection of wild-type CRC cells with mutant KRAS. The overexpression of Abi1 could be abolished by treatment with the PI3K-inhibitor Wortmannin after KRAS transfection.</p> <h3>Conclusions/Significance</h3><p>Our results support a role for Abi1 as a downstream target of inflammatory response and adenomatous change as well as oncogenic KRAS mutation via PI3K, but not BRAF activation. Furthermore, they highlight a possible role for Abi1 as a marker for early KRAS mutation in hyperplastic polyps. Since the protein is a key player in actin dynamics, our data encourages further studies concerning the exact role of Abi1 in actin reorganization upon enhanced KRAS/PI3K signalling during colonic tumorigenesis.</p> </div

Abi1 expression in analyzed samples.

<p>All values shown as mean ± SD; abbreviations: HPP: hyperplastic polyp; SSA/P: sessile serrated polyp/adenoma; TSA: traditional serrated adenoma; TbA: tubular adenoma; Ca: invasive colorectal carcinoma; Met: Metastasis; BRAF c600: B1 Rapidly accelerated fibrosarcoma codon 600 mutation; KRAS c12/13: Kirsten rat sarcoma codon 12/13 mutationMSI: microsatellite instable tumors; n: number of examined samples; n.a.: not applicable due to low sample number.</p

Clinico-pathologic characteristics of analyzed samples.

<p>Abbreviations: HPP: hyperplastic polyp; SSA/P: sessile serrated polyp/adenoma; TSA: traditional serrated adenoma; TbA: tubular adenoma; Ca: invasive colorectal carcinoma; Met: Metastasis; BRAF c600: B1 Rapidly accelerated fibrosarcoma codon 600 mutation; KRAS c12/13: Kirsten rat sarcoma codon 12 or 13 mutation.; n.t.: not tested; -: not applicable.</p

Abi1 expression analysis in specimens and cell lysates. A

<p>, Distribution of Abi1 expression in healthy and inflamed mucosa, hyperplastic polyps (HPP), sessile serrated polyps/adenomas (SSA/P), traditional serrated adenomas (TSA), tubular adenomas (TbA), invasive colorectal carcinoma (Ca) and metastases (Met). All values except BRAF-mutated TbA and carcinoma (each n = 1) are shown in box and whisker plot. Green squares represent maximum outliers, red squares represent minimum outliers. For inflamed mucosa, median, 1^st and 3^rd quartile are equal (score = 4). <b>B</b>, Statistical differences in Abi1 expression among all examined tissue specimens with respect to mutation status and, where applicable, microsatellite stability of each lesion. The lane for KRAS-mutated HPP is highlighted with a yellow background, the lane for KRAS-mutated invasive carcinoma is highlighted with a red background. The undermost line shows the number of examined samples in each group. <i>M: healthy mucosa; IM: inflamed mucosa; HP wt, HP K, HP B: wild-type, KRAS-mutated and BRAF-mutated hyperplastic polyps; SP wt, SP K, SP B: wild-type, KRAS-mutated and BRAF-mutated sessile serrated polyps/adenomas; TA wt, TA K: wild-type and KRAS-mutated traditional serrated adenomas; TbA wt, TbA K: wild-type and KRAS-mutated tubular adenomas; CA wt, CA K, CA MI: wild-type, KRAS-mutated and microsatellite-instable carcinomas; Met wt, Met K: wild-type and KRAS-mutated metastases; n.s.: not significant; *p<0.1;** p<0.05;*** p<0.01.</i></p

Proposed model for the regulation of actin dynamics via KRAS, PI3K and Abi1.

<p>Ligand-binding to membrane-associated receptor tyrosine kinase (RTK) leads to an activation of RAS, which is constitutively activated in mutant KRAS (*). Activated KRAS activates - among others - both the B1 Rapidly accelerated fibrosarcoma (BRAF)- and Phosphatidylinositol-3-kinase (PI3K)-pathway, only the latter leading to activation of the Abi1/Sos1/Eps8 complex. Via activation of the small GTPase Rac, this leads to reorganization of the actin cytoskeleton and to a change in cellular shape. <i>RTK: receptor tyrosine kinase; KRAS: Kirsten rat sarcoma; BRAF: B1 Rapidly accelerated fibrosarcoma</i>; <i>PI3K: Phosphatidylinositol-3-kinase; Abi1: Abelson interactor 1; Eps8: Epidermal growth factor receptor kinase substrate</i>; <i>Sos1: Son of sevenless homolog 1</i>; <i>Rac: Ras-related C3 botulinum toxin substrate.</i></p

Abi1 in colorectal cancer cell lines. A

<p>, Abi1 immunoblotting of colorectal carcinoma whole cell line lysates with different KRAS/BRAF mutation status shows upregulation of Abi1 in KRAS-mutated SW620 and SW1116, but only a faint signal in BRAF-mutated Colo205 cells. <b>B</b>, Immunoblotting of CHD-1 and HDC-9 cell lysates show overexpression of Abi1 in CHD1 cells, while both cell lines express comparable amounts of PI3K (I). There is slightly stronger Akt phosphorylation in CHD1 compared to HDC9 cells. Application of 50 nM Wortmannin (WO) almost completely repressed the pAkt and Abi1 signals. Immunofluorescence microscopy shows strong cytoplasmatic and nuclear Abi1 staining in CHD-1 cells, but only a faint cytoplasmatic signal in HDC-9 cells (II). <b>C,</b> KRAS/BRAF mutation testing reveals an activating KRAS G13D mutation in CHD-1 (left lane), while HDC-9 cells are KRAS wild-type (central lane). Transfection of HDC-9 cells with a KRAS G12D-construct leads to appearance of a band indicating a KRAS G12D-mutation (right lane). Both cell lines are BRAF wild-type. <b>D,</b> Immunoblotting of HDC-9 after transfection, TNFalpha and Wortmannin treatment shows an increase in pErk1/2 and pAkt and overexpression of Abi1 upon transfection with constitutively active KRAS G12D (2^nd lane) compared to both control (3^rd lane) and to transfection with wild-type KRAS (1^st lane). Stimulation with TNFalpha also enhances phosphorylation of signaling proteins and leads to upregulation of Abi1 (4^th lane). The overexpression of Abi1 could be reversed by application of 50 nM Wortmannin (5^th lane).</p