Comparison of HER2 and Phospho-HER2 Expression between Biopsy and Resected Breast Cancer Specimens Using a Quantitative Assessment Method

Background

HER2/Neu (ErbB-2) overexpression, which occurs in 15–20% of breast cancer cases, is associated with better response to treatment with the drug trastuzumab. PhosphoHER2 (pHER2) has been evaluated for prediction of response to trastuzumab. Both markers are heterogeneously detected and are potentially subject to loss as a consequence of delayed time to fixation. Here, we quantitatively assess both markers in core needle biopsies (CNBs) and matched tumor resections to assess concordance between the core and the resection and between HER2 and pHER2.

Methods

A selected retrospective collection of archival breast cancer cases yielded 67 cases with both core and resection specimens. Both HER2 and pTyr¹²⁴⁸HER2 were analyzed by the AQUA® method of quantitative immunofluorescence on each specimen pair.

Results

Both HER2 immunoreactivity (P<0.0001) and pTyr¹²⁴⁸HER2 immunoreactivity (P<0.0001) were lower in resections relative to CNB specimens. However, clinical implications of this change may not be evident since no case changed from 3+ (CNB) to negative (resection). Assessment of pTyr¹²⁴⁸HER2 showed no direct correlation with HER2 in either CNB or resection specimens.

Conclusions

The data suggest that measurement of both HER2 and phospho- Tyr¹²⁴⁸HER2, in formalin-fixed tissue by immunological methods is significantly affected by pre-analytic variables. The current study warrants the adequate handling of resected specimens for the reproducible evaluation of HER2 and pHER2. The level of pTyr¹²⁴⁸HER2, was not correlated to total HER2 protein. Further studies are required to determine the significance of these observations with respect to response to HER2 directed therapies.     

Oral and Craniofacial Manifestations and Two Novel Missense Mutations of the NTRK1 Gene Identified in the Patient with Congenital Insensitivity to Pain with Anhidrosis

Congenital insensitivity to pain with anhidrosis (CIPA) is a rare inherited disorder of the peripheral nervous system resulting from mutations in neurotrophic tyrosine kinase receptor 1 gene (NTRK1), which encodes the high-affinity nerve growth factor receptor TRKA. Here, we investigated the oral and craniofacial manifestations of a Chinese patient affected by autosomal-recessive CIPA and identified compound heterozygosity in the NTRK1 gene. The affected boy has multisystemic disorder with lack of reaction to pain stimuli accompanied by self-mutilation behavior, the inability to sweat leading to defective thermoregulation, and mental retardation. Oral and craniofacial manifestations included a large number of missing teeth, nasal malformation, submucous cleft palate, severe soft tissue injuries, dental caries and malocclusion. Histopathological evaluation of the skin sample revealed severe peripheral nerve fiber loss as well as mild loss and absent innervation of sweat glands. Ultrastructural and morphometric studies of a shed tooth revealed dental abnormalities, including hypomineralization, dentin hypoplasia, cementogenesis defects and a dysplastic periodontal ligament. Genetic analysis revealed a compound heterozygosity- c.1561T>C and c.2057G>A in the NTRK1 gene. This report extends the spectrum of NTRK1 mutations observed in patients diagnosed with CIPA and provides additional insight for clinical and molecular diagnosis.     

乏情和发情初产母猪下丘脑–垂体–卵巢轴中lincRNAs表达谱比较分析

初产母猪断奶后能否正常发情对养猪生产影响重大,也是初产母猪被淘汰的主要原因。本研究以乏情和发情初产母猪为研究对象,首次利用RNA-seq技术对其下丘脑-垂体-卵巢轴中的基因间长链非编码RNAs(long intergenic noncoding RNAs,lincRNAs)进行筛选比较,得到lincRNAs的表达图谱,并对其特征和功能进行了初步分析。结果显示,在乏情和发情初产母猪下丘脑–垂体–卵巢轴中鉴定得到3519个lincRNAs,以发情组为对照共有17个lincRNAs存在差异表达,其中12个表达上调,5个表达下调(FC≥2,P<0.05)。选择4个差异表达的lincRNAs经qRT-PCR验证,其表达水平与测序结果基本一致。对这17个差异表达的lincRNAs进行GO分析、KEGG通路分析及lincRNA-mRNA共表达网络分析,发现这些lincRNAs主要与猪卵母细胞减数分裂成熟、卵巢细胞分化及颗粒细胞凋亡等生殖活动相关。本研究结果丰富了猪lincRNAs数据资源,为进一步深入研究初产母猪的生殖机能提供了理论依据。     

Dietary adaptations and palaeoecology of Lophialetidae (Mammalia,Tapiroidea) from the Eocene of the Erlian Basin,China: combined evidence from mesowear and stable isotope analyses

Lophialetidae is an extinct group of endemic Asiatic tapiroids that are widely distributed in the Eocene sediments of Asia. Schlosseria magister and Lophialetes expeditus are the most abundant species in this family. However, their dietary and ecological characteristics are largely unknown. For the first time, we reconstruct the palaeodiet and habitat of these two lophialetids using a combination of mesowear and stable carbon isotope analysis of fossil teeth excavated from the Erlian Basin, China. Mesowear analysis (n = 141) suggests that the dietary structure of S. magister and L. expeditus shifted from less to more abrasive diets from ~52 to ~42 Ma. Stable carbon isotope analysis (n = 137) suggests that the habitats of S. magister and L. expeditus became drier and/or more open through time. The dietary shifts of the two lophialetids are consistent with evident changes in habitat. The changes in the diet and habitat were probably related to global climate change during that time period. The gradual drop in global temperatures during the early–middle Eocene led to a drier and more open terrestrial ecosystem in the Erlian Basin, probably resulting in changes in floral composition of the environment inhabited by S. magister and L. expeditus. Hence, herbivores highly susceptible to vegetation modification had to develop new resource exploitation strategies to adapt to these changes. Schlosseria magister, considered to be the sister-group of L. expeditus and with a low level of ecological flexibility, was unable to adapt to the habitat changes finally becoming extinct at ~45 Ma.     

Functional properties and the oligomeric state of alkyl hydroperoxide reductase subunit F (AhpF) in Pseudomonas aeruginosa

Protoplasma - Alkyl hydroperoxide reductase subunit F (AhpF) is a well-known flavoprotein that transfers electrons from pyridine nucleotides to the peroxidase protein AhpC via redox-active...     

Correction to: The Effects of 50 nm Unmodified Nano-ZnO on Lipid Metabolism and Semen Quality in Male Mice

Biological Trace Element Research - The original version of this article unfortunately contained a mistake. The correct title should be “The Effects of 50 nm Unmodified Nano-ZnO on Lipid...     

Integrated Bioinformatics Analysis for the Identification of Key Molecules and Pathways in the Hippocampus of Rats After Traumatic Brain Injury

Neurochemical Research - High-throughput and bioinformatics technology have been broadly applied to demonstrate the key molecules involved in traumatic brain injury (TBI), while no study has...     

High-efficiency protein delivery into transfection-recalcitrant cell types

Intracellular delivery of functional proteins is of great interest for basic biological research as well as for clinical applications. Transfection is the most commonly used method, however, it is not applicable to large-scale manipulation and inefficient in important cell types implicated in biomedical applications, such as epithelial, immune and pluripotent stem cells. In this study, we explored a bacterial type III secretion system (Bac-T3SS)-mediated proteofection method to overcome these limitations. An attenuated Pseudomonas aeruginosa vector was constructed, which has features of low toxicity, high T3SS activity, and self-limiting growth. Compared to the method of transfection, the Bac-T3SS showed significantly higher efficiencies of Cre recombinase translocation and target site recombination for hard-to-transfect human cell lines. Furthermore, through the delivery of β-lactamase in live animals, we demonstrated the feasibility and biosafety of in vivo application of the Bac-T3SS. This study provided an efficient and low-cost proteofection strategy for laboratory use as well as for application in large-scale cell manipulations.