The Histochemical Journal最新文献_第4页

Polymerase chain reaction in situ: an appraisal of an emerging technique. 原位聚合酶链反应:一项新兴技术的评价。

The Histochemical Journal Pub Date : 1995-09-01

I A Teo, S Shaunak

{"title":"Polymerase chain reaction in situ: an appraisal of an emerging technique.","authors":"I A Teo, S Shaunak","doi":"","DOIUrl":"","url":null,"abstract":"Polymerase chain reaction (PCR) in situ is a new technique which promises to enhance considerably our ability to detect a few copies of target nucleic acid sequences in fixed tissues and cells. It has an enormous potential for application in diagnostic histopathology of viral diseases and in the study of gene expression. PCR in situ is, however, technically difficult, and amplification of the target DNA is only 30-300 fold. In this article we present an overview of PCR in situ techniques used to amplify both DNA and RNA targets (RT-PCR in situ). We also identify problems which can reduce the efficiency of the technique or which can give rise to false-positive results. They include (1) the inhibitory effects of cross-linking of histones to DNA or PCR amplification, (2) abstraction of PCR reagents by tissue-bonding agents which are used to coat glass slides, (3) poor denaturation of target DNA and subsequent DNA renaturation due to extensive cross-linking of histones to DNA, or because of incorrect temperature regulation of thermal cyclers, (4) false-positive results which arise from end-labelling of DNA strand breaks by Taq polymerase, and (5) diffusion of PCR products into and out of cells leading to false-positive results. We present some of the approaches that have been used to overcome some of these difficulties and suggest new avenues for investigation to improve this technique further.","PeriodicalId":22439,"journal":{"name":"The Histochemical Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19536793","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

PCR in situ: aspects which reduce amplification and generate false-positive results. 原位PCR:减少扩增和产生假阳性结果的方面。

The Histochemical Journal Pub Date : 1995-09-01

I A Teo, S Shaunak

引用次数: 0

Expression of the CD15 antigen (Lewis x) in breast cancer. CD15抗原(Lewis x)在乳腺癌中的表达。

The Histochemical Journal Pub Date : 1995-09-01

S A Brooks, A J Leathem

引用次数: 0

The mATPase histochemical profile of rat type IIX fibres: correlation with myosin heavy chain immunolabelling. 大鼠IIX型纤维的mATPase组织化学特征:与肌球蛋白重链免疫标记的相关性。

The Histochemical Journal Pub Date : 1995-09-01

J A Santána Pereira, A de Haan, A Wessels, A F Moorman, A J Sargeant

{"title":"The mATPase histochemical profile of rat type IIX fibres: correlation with myosin heavy chain immunolabelling.","authors":"J A Santána Pereira, A de Haan, A Wessels, A F Moorman, A J Sargeant","doi":"","DOIUrl":"","url":null,"abstract":"In the present study we report a novel histochemical method which, by sequential pre-incubations in alkaline and acidic media, selectively differentiates muscle fibres expressing myosin heavy chain IIX, on the basis of a specific profile for myofibrillar actomyosin ATPase (mATPase) activity. The enzyme reactions were tested for specificity by means of anti-myosin heavy chain monoclonal antibodies, which were characterized on Western blots of muscle homogenates. Enzyme histochemical reactions with the traditional pH buffers were compared to those of the new method and, in conjunction with the immunoreactions, used to confirm the relationship between MyHC expression and the distinct profiles for mATPase. Immunohistochemical reactions demonstrated that the new method only differentiates those fibres expressing myosin heavy chain IIX. The method revealed a continuum in which the intermediate staining intensities corresponded to hybrid fibres expressing myosin heavy chain IIX in combination with either the IIA or IIB forms. Quantitative histochemistry and immunohistochemistry (by image analysis), used to examine the relationship between staining intensities for mATPase and amounts of myosin heavy chain IIX expression, revealed that the new method discriminates well between hybrid fibres expressing variable amounts of the IIX isoform (r2 = 0.93).","PeriodicalId":22439,"journal":{"name":"The Histochemical Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19538064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Histochemical study of glycoconjugates in the epididymis of the hamster (Mesocricetus auratus). 仓鼠附睾糖缀合物的组织化学研究。

The Histochemical Journal Pub Date : 1995-09-01

A Calvo, L M Pastor, R Horn, J Pallares

{"title":"Histochemical study of glycoconjugates in the epididymis of the hamster (Mesocricetus auratus).","authors":"A Calvo, L M Pastor, R Horn, J Pallares","doi":"","DOIUrl":"","url":null,"abstract":"The glycoconjugates of hamster epididymis were investigated with conventional and lectin histochemistry. A zone of the caput epididymis, with particular histochemical characteristics, has been differentiated. beta-Elimination in combination with lectins was used to establish the presence and distribution of N- and O-linked glycoconjugates. The epithelium, spermatozoa and the intertubular matrix were rich in glycoconjugates. The Golgi apparatus and stereocilia of the principal cells were intensely positive with HPA, PNA and SBA lectins. beta-Elimination indicated that these cells contained abundant O-linked glycoconjugates. Apical and clear cells presented a common lectin affinity; their reactivities towards WGA and UEA-I were very positive. These cells probably contain abundant N-glycoconjugates. The spermatozoa were stained by periodic acid-Schiff (PAS) and by all the lectins (especially in the acrosome), except by those with an affinity for alpha-L-fucosyl residues; the most intense reaction was found with HPA, WGA, PNA and SBA. Changes in the sperm lectin binding along the ductus were observed: sperm flagellum abruptly acquired WGA and PNA labelling from the posterior caput, and HPA reactivity was negative only in the zone between the caput and the corpus.","PeriodicalId":22439,"journal":{"name":"The Histochemical Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19536795","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A cytophotometric and electron-microscopical study on catalase activity in serial cryostat sections of rat liver. 大鼠肝脏连续低温切片过氧化氢酶活性的细胞光度和电镜研究。

The Histochemical Journal Pub Date : 1995-09-01

W M Frederiks, M Ankum, K S Bosch, H Vreeling-Sindelárová, J P Schellens, C J Van Noorden

{"title":"A cytophotometric and electron-microscopical study on catalase activity in serial cryostat sections of rat liver.","authors":"W M Frederiks, M Ankum, K S Bosch, H Vreeling-Sindelárová, J P Schellens, C J Van Noorden","doi":"","DOIUrl":"","url":null,"abstract":"The validity of the histochemical procedure for demonstrating catalase activity in cryostat sections of rat liver at the light- and electron-microscopical level was studied cytophotometrically. Incubations in the presence of 5 mM diaminobenzidine, 44 mM hydrogen peroxide and 2% polyvinyl alcohol performed on fixed cryostat sections resulted in the highest amounts of final reaction product precipitated in a fine granular form which was specific for catalase activity. Serial sections processed for electron microscopy indicated that the osmiophilic final reaction product was exclusively localized in the matrix and core of peroxisomes. The relationship between incubation time and the amounts of final reaction product generated by catalase activity as measured at 460 nm in mid-zonal areas of liver lobules showed non-linearity for the test-minus-control reaction because first-order inactivation of the enzyme occurred during incubation. Linearity of the test-minus-control reaction and section thickness was observed up to 8 microns. Catalase in rat liver showed a Km value of 2.0 mM for its substrate hydrogen peroxide when the diaminobenzidine concentration was 5 mM. It is concluded that the procedure for demonstrating catalase activity in serial cryostat sections of rat liver at the light- and electron-microscopical level is specific and can be applied to quantitative purposes. This approach may be useful in pathology, when only small biopsies are available, when the tissue is heterogeneous, and when other histochemical markers also need to be studied in the same material.","PeriodicalId":22439,"journal":{"name":"The Histochemical Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19536796","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Immunohistochemical distribution of S-100 protein and type IV collagen in human embryonic and fetal sympathetic neuroblasts. S-100蛋白和IV型胶原在人胚胎和胎儿交感神经母细胞中的免疫组织化学分布。

The Histochemical Journal Pub Date : 1995-09-01

G Magro, S Grasso, C Emmanuele

{"title":"Immunohistochemical distribution of S-100 protein and type IV collagen in human embryonic and fetal sympathetic neuroblasts.","authors":"G Magro, S Grasso, C Emmanuele","doi":"","DOIUrl":"","url":null,"abstract":"The expression and distribution of S-100 protein and type IV collagen was studied immunohistochemically in sympathetic neuroblasts from the paravertebral region to the adrenal glands in human embryos and fetuses ranging from 7 to 12 weeks gestational age. From 7 weeks gestational age, S-100 protein was detected in round or oval cells mingling with sympathetic neuroblasts, and in spindle-shaped cells forming a continuous layer around them. The latter S-100 protein-positive cells were found in contact with the Schwann cells of nerve fibres entering the groups of sympathetic neuroblasts. Staining for type IV collagen showed that all groups of sympathetic neuroblasts were surrounded by a continuous basement membrane. By examining serial sections stained for type IV collagen and S-100 protein, a continuous basement membrane was found along the distribution pattern of the peripheral S-100 protein-positive spindle cells. The morphology of these cells, and their relationships with Schwann cells and with the basement membrane of the sympathetic neuroblasts, indicated that they were Schwann-like cells probably capable of synthesizing a continuous basement membrane separating the neuroblasts from the adjacent tissues. In contrast, the round or oval S-100 protein-positive cells, in contact with the sympathetic neuroblasts and not associated with nerve fibres, were considered as sustentacular or sustentacular precursor cells. At week 7 gestational age, the peri-adrenal sympathetic neuroblasts and their sustentacular and Schwann-like cells started to invade the adrenal glands and mingled with the adrenal cortical cells. These findings suggest the extra-adrenal origin of the sustentacular cells in embryonic and fetal adrenal glands.","PeriodicalId":22439,"journal":{"name":"The Histochemical Journal","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"1995-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19536798","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

In situ localization with digoxigenin-labelled probes of tau-related mRNAs in the rat pancreas. 用地高辛标记探针原位定位大鼠胰腺中tau相关mrna。

The Histochemical Journal Pub Date : 1995-08-01

P Neuville, M T Vanier, L Michalik, J F Launay

引用次数: 0

Effects of fixation on the preservation of peroxisomal structures for immunofluorescence studies using HepG2 cells as a model system. 用HepG2细胞作为模型系统进行免疫荧光研究，固定对过氧化物酶体结构保存的影响。

The Histochemical Journal Pub Date : 1995-08-01

M Schrader, E Baumgart, H D Fahimi

引用次数: 0

Carbonic anhydrase is present in human oesophageal epithelium and submucosal glands. 碳酸酐酶存在于人食管上皮和粘膜下腺。

The Histochemical Journal Pub Date : 1995-08-01

K N Christie, C Thomson, S Morley, J Anderson, D Hopwood

引用次数: 0