Computer-assisted image analysis of the distributions of peptidergic terminals in the central nucleus of the amygdala: A preliminary study

When viewed under dark-field illumination, peptidergic terminals in sections stained by the Sternberger PAP immunocytochemical method are seen as individual points of light. Under high magnification, the degree of brightness of various areas of immunoreactive terminals is seen to be a function of the density of terminals in these areas. By analyzying the relative brightness of the immunostained central nucleus of the amygdala (CNA) with an EyeCom II PDP-$\text{11}\text{34}$ image analysis system, we have obtained a relative evaluation of the density distribution of neurotensin (NT)-, substance P (SP), VIP-, angiotensin II (AII), m-enkephalin (m-ENK) and somatostatin (SS)-immunoreactive terminals in terms of normal morphology and following a brain lesion. The EyeCom II system divides the presented image into 307200 picture elements (pixels) and assigns one of 256 grey values to the average brightness with each pixel. We have aggregated the grey level frequencies into 5 levels where level 1 corresponds to the highest terminal density and level 5 to the lowest density. At level 1, only NT- and VIP-immunoreactive terminals occupy a significant percentage of the cross-sectional area of the CNA (20%). About 15% of the area of the CNA has VIP terminals with level 5 density. The distributions of the top 20% of the terminal density range of NT, SP, AII and VIP support a classical medial/lateral division of the nucleus. The distribution of the same range of SS- and ENK terminals suggests a dorsoventral division of the CNA. A preliminary study indicates that comparison of grey level frequency histograms generated by image analysis from homologous lesioned and unlesioned sections of the CNA can yield useful information regarding post-lesion changes in the distribution of immunoreactive terminals.     

Quantified 123I-Thyroid Scan based classification of hyperthyroidism

The original thyroid scan (TS) was widely used to identify typical imaging patterns, suggesting the widely accepted main following clinical diagnoses: Grave's disease, Toxic adenoma, [hetero]-nodular goiters and thyroiditis. With the diffusion of sensitive TSH assays, considerable advances in the comprehension of the molecular mechanisms of hormonosynthesis, and new quantification possibilities especially using ¹²³I, the TS is a textbook of molecular imaging. The image can be finely quantified with, not only as regards the Uptake (¹²³IUp) and related parameters but also, the quantification of the spatial targeting leading to a Spatial Target Index (STI). Using this new molecular ¹²³I-TS, TSH values, and when required, correlation to Multiparametric Ultrasounds (MPUS), we generated a basic classification system of hyperthyroidism, with well-defined indexed criteria (C11-1 to C17-3), that allows reporting 24 distinct etiologies. Selected criteria involve TS and contrast patterns, precocious ¹²³IUp (p¹²³IUp), maximal TSH-dependent physiological Uptake, lobar concentration, Uptake and concentration ratios, STI, ^99mTc-MIBI TS and correlative MPUS. This approach allows to identify 4 subtypes of Graves’ disease, including hyperplastic, nodular and common GD variants entangled with Hashimoto's struma, 4 subtypes of Thyroid Functional Autonomy, including Disseminated Functional Autonomy, that cannot be diagnosed with other conventional procedures. Criteria C14-1 to C17-3 report on hyperthyroidism and iodine overload, factitia, main thyroiditis presentations and rare central or tumoral etiologies of hyperthyroidism. This classification, based on ¹²³I-TS molecular imaging, leads to unprecedented diagnostic finesse and paves the way for a personalized theranostic approach in thyroid pathology. Further development towards artificial intelligence networks is under study.     

A new insight into the impact of different proteases on SILAC quantitative proteome of the mouse liver

In this study, we examined the use of multiple proteases (trypsin, LysC, tandem LysC/trypsin) on both protein identification and quantification in the Lys‐labeled SILAC mouse liver. Our results show that trypsin and tandem LysC/trypsin digestion are superior to LysC in peptides and protein identification while LysC shows advantages in quantification of Lys‐labeled proteins. Combination of experimental results from different proteases (LysC and trypsin) enabled a significant increase in the number of identified protein and protein can be quantified. Thus, taking advantage of the complementation of different protease should be a good strategy to improve both qualitative and quantitative proteomics research.     

基于电镜和免疫组化的线粒体数目标志物评估肝癌的初步研究

目的:研究线粒体数目与肝癌生长的相关性。方法:利用电子显微镜技术定量肝癌组织中线粒体数目,利用免疫组织化学技术评估肝癌组织中的线粒体标志物COX Ⅳ及HSP60表达水平,分析电镜肝癌线粒体定量结果与COX Ⅳ及HSP60表达量之间的相关性,并比较肝癌中线粒体数目、COX Ⅳ及HSP60表达量与肝癌直径之间的关系。结果:与癌旁相比,肝癌组织中线粒体数目(P0.001)、COX Ⅳ及HSP60的表达量显著降低(P=0.0417,P=0.0290)。COX Ⅳ及HSP60表达水平与线粒体数目无显著相关(r~2=0.009,P=0.5468;r~2=0.056,P=0.1396)。线粒体数目与肿瘤直径显著负相关(r~2=0.1086,P=0.0434),COX Ⅳ及HSP60表达量与肿瘤直径无显著相关(r~2=0.0251,P=0.3287;r~2=0.0461,P=0.1830)。结论:线粒体数目是潜在的肝癌生长标志物。但常用的线粒体定量分子HSP60与COX Ⅳ并不能准确定量肝癌线粒体。     

Substantial Influence of ERAP2 on the HLA-B*40:02 Peptidome: Implications for HLA-B*27-Negative Ankylosing Spondylitis

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Highlights

• •HLA-B*40:02 and ERAP2 are risk factors for ankylosing spondylitis.
• •The effects of ERAP2 on the B*40:02 peptidome are defined.
• •ERAP2 has a major influence mainly due to alterations of N-terminal residues.
• •These effects provide a basis for the association of ERAP2 with disease.

     

Bayesian Confidence Intervals for Multiplexed Proteomics Integrate Ion-statistics with Peptide Quantification Concordance,

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Highlights

• •Bayesian Beta-Binomial model integrates ion statistics with peptide ratio agreement.
• •Model appropriately interprets information from low signal peptides.
• •Confidence can be assigned even without replicates.
• •Model adds sensitivity to detection of small changes.

     

Insulin Induces Microtubule Stabilization and Regulates the Microtubule Plus-end Tracking Protein Network in Adipocytes

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Highlights

• •Insulin Affects the Phosphorylation of G2L1, MARK2, CLIP2, EB1, AGAP3, and CKAP5.
• •Insulin Increases CLASP2 +TIP Density and Decreases CLASP2 +TIP Velocity.
• •Insulin Stimulates CLASP2 and G2L1 Trailing Along Microtubules.
• •Insulin Stimulates α-Tubulin Acetylation at Lysine 40 and Microtubule Stabilization.