Ultrastructural and immunohistochemical study of early repair of alveolar sockets after the extraction of molars from alendronate‐treated rats

The aim of the present research was to analyze ultrastructural and immunohistochemical aspects of the alveolar repair after the extraction of molars of alendronate (ALN)‐treated rats. Wistar rats received 2.5mg/kg body wt/day of ALN during 14 days previously and 7, 14 and 21 days after the extraction of the second mandibular molar. Specimens were fixed in 2% glutaraldehyde + 2.5% formaldehyde under microwave irradiation, decalcified in 4.13% EDTA and paraffin embedded for TRAP histochemistry and immunohistochemistry for OPN, BSP and endoglin, or embedded in Spurr epoxy resin for TEM analysis. Additional specimens had their soft tissues removed and were processed for scanning electron microscopy. The ALN group presented latent TRAP‐positive osteoclasts and nonresorbed alveolar crests with bacterial infection. Mild bone necrosis signs were observed at all time points studied. Ultrastructurally, empty osteocyte lacunae were observed and bone trabeculae surface presented hyalinized aspect. A significant delay in alveolar repair occurred, as well as decreased angiogenesis. ALN treatment provoked mild signs of bone necrosis, despite the high dose employed. The present findings add new information about the ultrastructural aspect of the early repair of rats under ALN treatment and highlight for giving attention when oral surgeries are performed in patients using this drug. Microsc. Res. Tech. 76:633–640, 2013. © 2013 Wiley Periodicals, Inc.     

Immunohistochemical Evaluation of Adaptor Protein FAM159B Expression in Normal and Neoplastic Human Tissues

FAM159B is a so-called adaptor protein. These proteins are essential components in numerous cell signalling pathways. However, little is known regarding FAM159B expression in normal and neoplastic human tissues. The commercially available rabbit polyclonal anti-human FAM159B antibody HPA011778 was initially characterised for its specificity using Western blot analyses and immunocytochemistry and then applied to a large series of formalin-fixed, paraffin-embedded normal and neoplastic human tissue samples. Confirmation of FAM159B’s predicted size and antibody specificity was achieved in BON-1 cells, a neuroendocrine tumour cell line endogenously expressing FAM159B, using targeted siRNA. Immunocytochemical experiments additionally revealed cytoplasmic expression of the adaptor protein. Immunohistochemical staining detected FAM159B expression in neuronal and neuroendocrine tissues such as the cortex, the trigeminal ganglia, dorsal root and intestinal ganglia, the pancreatic islets and the neuroendocrine cells of the bronchopulmonary and gastrointestinal tract, but also in the syncytiotrophoblasts of the placenta. FAM159B was also expressed in many of the 28 tumour entities investigated, with high levels in medullary and anaplastic thyroid carcinomas, parathyroid adenomas, lung and ovarian carcinomas, lymphomas and neuroendocrine tumours of different origins. The antibody HPA011778 can act as a useful tool for basic research and identifying FAM159B expression in tissue samples.     

Correlation of MET-Receptor Overexpression with MET Gene Amplification and Patient Outcome in Malignant Mesothelioma

Thanks to clinically newly introduced inhibitors of the mesenchymal–epithelial transition (MET) receptor tyrosine-kinase, MET-gene copy number gain/amplification (MET-GCNG/GA) and increased expression of the MET protein are considered very promising therapeutic targets in lung cancer and other malignancies. However, to which extent these MET alterations occur in malignant mesothelioma (MM) remains unclear. Thus, we investigated by well-established immunohistochemistry and fluorescence in situ hybridization methods, the frequency of these alterations in specimens from 155 consecutive MMs of different subtypes obtained from pleural or peritoneal biopsies and pleurectomies. Thirty-three benign reactive mesothelial proliferations (RMPs) were used as controls. MET-protein upregulation was observed in 35% of all MM-cases, though restricted to predominantly epithelioid MMs. We detected low-/intermediate-level MET-GCNG/GA in 22.2% of MET-overexpressing MMs (7.8% of whole MM-cohort) and no MET-GCNG/GA in the other 77.8%, suggesting other upregulating mechanisms. In contrast, 100% of RMPs exhibited no MET-upregulation or MET-GCNG/-GA. Neither MET exon 14 skipping mutations nor MET-fusions were detected as mechanisms of MET overexpression in MM using RNA next-generation sequencing. Finally, in two cohorts of 30 MM patients with or without MET overexpression (MET-positive/-negative) that were matched for several variables and received the same standard chemotherapy, the MET-positive cases showed a significantly lower response rate, but no significant difference in progression-free or overall survival. Our results imply that MET overexpression occurs in a substantial fraction of predominantly epithelioid MMs, but correlates poorly with MET-amplification status, and may impact the likelihood of response to mesothelioma standard chemotherapy. The predictive significance of MET-IHC and -FISH for possible MET-targeted therapy of MM remains to be elucidated.     

Peptides in frog brain areas processing visual information

Vision is the most important sensory modality to anurans and a great deal of work in terms of hodological, physiological, and behavioral studies has been devoted to the visual system. The aim of this account is to survey data about the distribution of peptides in primary (lateral geniculate complex, pretectum, tectum) and secondary (striatum, anterodorsal and anteroventral tegmental nuclei, isthmic nucleus) visual relay centers. The emphasis is on general traits but interspecies variations are also noted. The smallest amount of peptide-containing neuronal elements was found in the lateral geniculate complex, where primarily nerve fibers showed immunostaining. All peptides found in the lateral geniculate complex, except two, occurred in the pretectum together with four other peptides. A large number of neurons showing intense neuropeptide thyrosine-like immunoreactivity was characteristic here. The mesencephalic tectum was the richest in peptide-like immunoreactive neuronal elements. Almost all peptides investigated were present mainly in fibers, but 9 peptides were found also in cells. The immunoreactive fibers show a complicated overlapping laminar arrangement. Cholecystokinin octapeptide, enkephalins, neuropeptide tyrosine, and substance P (not discussed here) gave the most prominent immunoreactivity. Several peptides also occur in the tectum of fishes, reptiles, birds, and mammals. Peptides in various combinations were found in the striatum, the anterodorsal- and anteroventral tegmental nucleus, and the isthmic nucleus that receive projections from the primary visual centers. The functional significance of peptides in visual information processing is not known. The only exception is neuropeptide tyrosine, which was found to be inhibitory on retinotectal synapses.     

胃肠道间质瘤的电镜和免疫组织化学形态特征

应用光学显微镜、电子显微镜和免疫组织化学方法,观察分析16例胃肠道间质瘤（GIST）的肿瘤分化形态特征和免疫表达。16例GIST电镜观察可查见各种分化的特征性超微结构：SF（skeinoid fibers）神经内分泌颗粒、Luse小体（长间跨胶原）、微管、突触样结构及平滑肌特征性结构,显示平滑肌瘤、神经鞘瘤和神经源性的特征性结构。免疫组织化学染色显示16例GIST中S－100蛋白、SMA和CD34的阳性率分别为81.3%、87.5%和87.5%;SMA、S－100和CD34三项均阳性9例,SMA和S－100阳性2例,SMA和CD34阳性3例,S－100、SMA阳性2例。超微结构和免疫组织化学观察结果表明,GIST呈现多向分化和混合表达,提示GIST起源于源始间叶细胞,而小肠间质瘤则大多数为神经源性肿瘤。电镜结合光学显微镜、免疫组织化学检查,对GIST的诊断和判断预后有辅助作用。     

Comparative study of DNA synthesis and nucleolar organizer regions of sinusoid littoral cells in mouse regenerating liver.

Variations in DNA synthesis (DNAs) and Nucleolar Organizer Regions (NORs) were studied in the littoral cell population from regenerating liver of C3HS inbred mice standardized for periodicity analysis. Immunohistochemical detection of Bromodeoxyuridine (BrdU) with a monoclonal antibody and silver staining of NORs (AgNORs) were assessed by means of a digital image analysis system in histological sections. Tissue samples were obtained every four hours from the 30th to the 54th hours after a partial hepatectomy. The results showed, in both parameters, a gradual increment of the values during the period studied, with highest values (DNAs 107.1 +/- 16.1 SE; AgNORs 77.3 +/- 3.4 SE) located at 16:00/54 Time of Day / Hours Post-Hepatectomy (TD/HPH), which were significantly different (p <0.001) from the values of the first sample (DNAs 38.1 +/- 9.5 SE; AgNORs 27.3 +/- 1.0 SE) taken at 16:00/30 TD/HPH. The results of our experiment demonstrate the existence of a strong correlation of DNA synthesis measured by BrdU immunohistochemistry and AgNORs numbers in sinusoid littoral cells from mouse regenerating liver.     

30例乳腺导管内癌诊断方法的探讨

目的：探讨彩色多普勒超声、X 线钼靶、术中冰冻检查、常规病理检查及免疫组织化学检测对乳腺导管内癌（ductal carcinoma in situ,DCIS）患者术前、术中、术后的诊断价值。方法对30例 DCIS 患者的临床资料,X 线钼靶、彩色多普勒超声、术中冰冻检查结果及 CK5／6、CK8／18、34βE12、Ki67等免疫组织化学检测结果进行回顾性分析。结果30例 DCIS 患者中症状为乳腺肿块或乳腺局限性增厚23例、单独或伴发乳头溢液5例、不随月经周期而改变的乳腺疼痛1例、没有任何症状1例。彩色多普勒超声检测阳性符合率为66．7％,X 线钼靶检测阳性符合率为76．7％,2种检测方法比较差异无统计学意义（χ2＝0．82,P ＞0．05）;彩色多普勒超声联合 X 线钼靶检测阳性符合率明显高于彩色多普勒超声单独检测阳性符合率（90．0％比66．7％,χ2＝4．812,P ＜0．05）。术中冰冻 DCIS检出率为66．7％。CK5／6在 DCIS 及乳脉导管不典型增生（ADH）区域几乎为阴性,仅1例增生导管上皮 CK5／6及 CK8／18呈阳性;4例34βE12在 ADH 和 DCIS 增生细胞中均有表达;15例 CK8／18在常规组织学病理中明确的DCIS 区域及 ADH 区域中呈阳性表达;Ki67在 DCIS 区域中的表达较 ADH 区域略有增高趋势。结论彩色多普勒超声、X 线钼靶、术中冰冻检查、常规病理检查及免疫组织化学检测对 DCIS 患者术前、术中、术后诊断各有优点及局限性,临床医生应综合考虑有利于 DCIS 的诊断,减少漏诊。