The Immunohistochemical Method

The objective: to еxplore the relationship between the activity of endometrial proliferation and the state of the local immune response in the uterus in the conditions berprestasi process. Patients and methods. Examined 228 women of reproductive and perimenopausal age with endometrial pathology using ultrasound and then performing hysteroresectoscopy. Determination of the concentrations of the cytokines IL-1, IL-2, IL-6 and TNF was performed by solid phase ELISA. Results. Found a trend that confirms the loss of sensitivity to hormones at the stage of malignancy of the endometrium and can be used as diagnostic determinants in determining the nature of intrauterine pathology and criterion of the effectiveness of conservative therapy. Conclusion. Improving etiopatogenetice approach to the therapy of hyperplastic proce.sses of endometrium with determination of receptor phenotype of the endometrium is a research direction in modern gynecology, which will help to improve the results of treatment and prevention of intrauterine pathology. Key words: endometrial hyperplasia,the receptors for progesterone and estrogen, immunohistochemical method.

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C-fos, ERK1/2, MAP2, NOTCH1 Proteins Expression Patterns in Human Cerebral Cortex Neurons after Ischemic Stroke

Annals of the Russian academy of medical sciences ◽

10.15690/vramn1295 ◽

2020 ◽

Vol 75 (3) ◽

pp. 226-233

Author(s):

Svetlana P. Sergeeva ◽

Aleksey V. Lyundup ◽

Valery V. Beregovykh ◽

Petr F. Litvitskiy ◽

Aleksey A. Savin ◽

...

Keyword(s):

Cerebral Cortex ◽

Ischemic Stroke ◽

Expression Patterns ◽

Immunohistochemical Method ◽

Left Middle Cerebral Artery ◽

Contralateral Hemisphere ◽

Human Cerebral Cortex ◽

Fos Protein ◽

Urgent Task ◽

Indirect Immunoperoxidase

Background. The search for protein (these include c-fos, ERK1/2, MAP2, NOTCH1) expression that provide neuroplasticity mechanisms of the cerebral cortex after ischemic stroke (IS) patterns is an urgent task. Aims to reveal c-fos, ERK1/2, MAP2, NOTCH1 proteins expression patterns in human cerebral cortex neurons after IS. Materials and methods. We studied 9 left middle cerebral artery (LMCA) IS patients cerebral cortex samples from 3 zones: 1 the zone adjacent to the necrotic tissue focus; 2 zone remote from the previous one by 47 cm; 3 zone of the contralateral hemisphere, symmetric to the IS focus. Control samples were obtained from 3 accident died people. Identification of targeted proteins NSE, c-fos, ERK1/2, MAP2, NOTCH1 was performed by indirect immunoperoxidase immunohistochemical method. Results. Moving away from the ischemic focus, there is an increase in the density of neurons and a decrease in the damaged neurons proportion, the largest share of c-fos protein positive neurons in zone 2, NOTCH1 positive neurons in zone 1, smaller fractions of ERK1/2 and MAP2 positive neurons compared to the control only in samples of zone 1. Conclusions. With the IS development, the contralateral hemisphere is intact tissue increased activation zone, while the zones 1 and 2 have pathological activation signs. In zone 1 of the range, the adaptive response of the tissue decreases, and in zone 2 it expands. Therefore, a key target for therapeutic intervention is zone 2.

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The Telocytes in the Subepicardial Niche

Applied Sciences ◽

10.3390/app9081615 ◽

2019 ◽

Vol 9 (8) ◽

pp. 1615 ◽

Cited By ~ 5

Author(s):

Cristian Bogdan Iancu ◽

Mugurel Constantin Rusu ◽

Laurenţiu Mogoantă ◽

Sorin Hostiuc ◽

Oana Daniela Toader

Keyword(s):

Stromal Cells ◽

Lymphatic Vessels ◽

Growth Factor Receptor ◽

Immunohistochemical Method ◽

Lymphatic Endothelial Cells ◽

Cell Type ◽

Ki 67 ◽

Epicardial Cells ◽

Immunohistochemical Evidence ◽

Unique Cell

A great interest has developed over the last several years in research on interstitial Cajal-like cells (ICLCs), later renamed to telocytes (TCs). Such studies are restricted by diverse limitations. We aimed to critically review (sub)epicardial ICLCs/TCs and to bring forward supplemental immunohistochemical evidence on (sub)epicardial stromal niche inhabitants. We tested the epicardial expressions of CD117/c-kit, CD34, Cytokeratin 7 (CK7), Ki67, Platelet-Derived Growth Factor Receptor (PDGFR)-α and D2-40 in adult human cardiac samples. The mesothelial epicardial cells expressed D2-40, CK7, CD117/c-kit and PDGFR-α. Subepicardial D2-40-positive lymphatic vessels and isolated D2-40-positive and CK7-positive subepicardial cells were also found. Immediate submesothelial spindle-shaped cells expressed Ki-67. Submesothelial stromal cells and endothelial tubes were PDGFR-α-positive and CD34-positive. The expression of CD34 was pan-stromal, so a particular stromal cell type could not be distinguished. The stromal expression of CD117/c-kit was also noted. It seems that epicardial TCs could not be regarded as belonging to a unique cell type until (pre)lymphatic endothelial cells are inadequately excluded. Markers such as CD117/c-kit or CD34 seem to be improper for identifying TCs as a distinctive cell type. Care should be taken when using the immunohistochemical method and histological interpretations, as they may not produce accurate results.

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Distribution of substance P in the spinal cord of patients with syringomyelia

Journal of Neurosurgery ◽

10.3171/jns.1996.84.6.0992 ◽

1996 ◽

Vol 84 (6) ◽

pp. 992-998 ◽

Cited By ~ 10

Author(s):

Thomas H. Milhorat ◽

Harrison T. M. Mu ◽

Carole C. LaMotte ◽

Ade T. Milhorat

Keyword(s):

Spinal Cord ◽

Substance P ◽

Dorsal Horn ◽

Ventral Horn ◽

Immunohistochemical Method ◽

Positive Staining ◽

Content Type ◽

Normal Individuals ◽

Neurologically Normal ◽

Intermediolateral Nucleus

✓ The distribution of substance P, a putative neurotransmitter and pain-related peptide, was studied using the peroxidase—antiperoxidase immunohistochemical method in the spinal cords obtained from autopsy of 10 patients with syringomyelia and 10 age- and sex-matched, neurologically normal individuals. Substance P immunoreactivity was present in axons and in terminal-like processes in close apposition to neurons in the first, second, and third laminae of the dorsal horn. Smaller amounts of peroxidase-positive staining were found in the fifth lamina of the dorsal horn, the intermediolateral nucleus, the intermediomedial nucleus, and the ventral horn. In nine of 10 patients with syringomyelia, there was a substantial increase in substance P immunoreactivity in the first, second, third, and fifth laminae below the level of the lesion. A marked reduction or absence of staining was present in segments of the spinal cord occupied by the syrinx. Central cavities produced bilateral abnormalities, whereas eccentric cavities produced changes that were ipsilateral to the lesion. No alterations in staining were found in the spinal cord of an asymptomatic patient with a small central syrinx. The authors conclude that syringomyelia can be associated with abnormalities in spinal cord levels of substance P, which may affect the modulation and perception of pain.

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Immunohistochemical localization of pituitary gonadotrophins and gonadal steroids confirms the 'two-cell, two-gonadotrophin' hypothesis of steroidogenesis in the human ovary

Journal of Endocrinology ◽

10.1677/joe.0.1260483 ◽

1990 ◽

Vol 126 (3) ◽

pp. 483-NP ◽

Cited By ~ 35

Author(s):

M. Kobayashi ◽

R. Nakano ◽

A. Ooshima

Keyword(s):

Luteal Phase ◽

Immunohistochemical Analysis ◽

Immunohistochemical Localization ◽

Follicular Phase ◽

Gonadal Steroids ◽

Immunohistochemical Method ◽

Human Ovary ◽

Luteal Cells ◽

Menstrual Cycles

ABSTRACT Ovaries from 37 women with normal menstrual cycles were analysed for localization of pituitary gonadotrophins and gonadal steroids using an immunohistochemical method. In the follicular phase, FSH and oestradiol-17β localized in the granulosa layer, and LH, progesterone and testosterone localized in the internal thecal layer. In the luteal phase, gonadotrophins and steroids localized in luteal cells. Particularly in the early luteal phase, FSH and oestradiol-17β localized in large luteal cells, and LH, progesterone and testosterone localized in small luteal cells. The results of the present immunohistochemical analysis confirm the two-cell, two-gonadotrophin hypothesis of steroidogenesis in the human ovary. Journal of Endocrinology (1990) 126, 483–488

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The Role of EGFR/PI3K/Akt/cyclinD1 Signaling Pathway in Acquired Middle Ear Cholesteatoma

Mediators of Inflammation ◽

10.1155/2013/651207 ◽

2013 ◽

Vol 2013 ◽

pp. 1-9 ◽

Cited By ~ 16

Author(s):

Wei Liu ◽

Hongmiao Ren ◽

Jihao Ren ◽

Tuanfang Yin ◽

Bing Hu ◽

...

Keyword(s):

Cell Cycle ◽

Cell Proliferation ◽

Signaling Pathways ◽

Signaling Pathway ◽

External Auditory Canal ◽

Cell Cycle Progression ◽

Critical Role ◽

Immunohistochemical Method ◽

Cycle Progression ◽

Middle Ear Cholesteatoma

Cholesteatoma is a benign keratinizing and hyper proliferative squamous epithelial lesion of the temporal bone. Epidermal growth factor (EGF) is one of the most important cytokines which has been shown to play a critical role in cholesteatoma. In this investigation, we studied the effects of EGF on the proliferation of keratinocytes and EGF-mediated signaling pathways underlying the pathogenesis of cholesteatoma. We examined the expressions of phosphorylated EGF receptor (p-EGFR), phosphorylated Akt (p-Akt), cyclinD1, and proliferating cell nuclear antigen (PCNA) in 40 cholesteatoma samples and 20 samples of normal external auditory canal (EAC) epithelium by immunohistochemical method. Furthermore,in vitrostudies were performed to investigate EGF-induced downstream signaling pathways in primary external auditory canal keratinocytes (EACKs). The expressions of p-EGFR, p-Akt, cyclinD1, and PCNA in cholesteatoma epithelium were significantly increased when compared with those of control subjects. We also demonstrated that EGF led to the activation of the EGFR/PI3K/Akt/cyclinD1 signaling pathway, which played a critical role in EGF-induced cell proliferation and cell cycle progression of EACKs. Both EGFR inhibitor AG1478 and PI3K inhibitor wortmannin inhibited the EGF-induced EGFR/PI3K/Akt/cyclinD1 signaling pathway concomitantly with inhibition of cell proliferation and cell cycle progression of EACKs. Taken together, our data suggest that the EGFR/PI3K/Akt/cyclinD1 signaling pathway is active in cholesteatoma and may play a crucial role in cholesteatoma epithelial hyper-proliferation. This study will facilitate the development of potential therapeutic targets for intratympanic drug therapy for cholesteatoma.

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COMPARISON OF CLINICOPATHOLOGICAL PARAMETERS WITH HISTOLOGICAL AND MOLECULAR CLASSIFICATION OF BREAST CANCERS

10.36106/gjra/4912531 ◽

2021 ◽

pp. 144-149

Author(s):

K. Indumathi ◽

E. Theranirajan ◽

G. Bhavani

Keyword(s):

Retrospective Study ◽

Targeted Therapy ◽

Tertiary Care ◽

Radical Mastectomy ◽

Immunohistochemical Method ◽

Proliferation Index ◽

Clinicopathological Parameters ◽

P Value ◽

Luminal B ◽

Classi Cation

BACKGROUND: This is a retrospective study of 60 cases, to detect the expression of ER, PR, HER2neu, CK5/6 and Ki67 proliferation index in breast carcinomas by immunohistochemical method and to determine the newer molecular classication. Few patients have recurrence inspite of being diagnosed under the category of low risk and few do well in the high risk group which can be attributed to the molecular level differentiation. AIM: The aim of this study is to categorize the patients under molecular classication, and to compare the clinicopathological parameters with it and to denote the signicance of targeted therapy. MATERIALS AND METHODS: A retrospective study of detecting the expression of the above said markers in modied radical mastectomy specimens received at a tertiary care centre during the period from January 2015 to June 2018. A total of 60 cases which included 30 of IDC NST and 30 cases of special variants were selected for immunohistochemical analysis. RESULTS: Out of the 60 cases studied, the most common was found to be the luminal A type comprising 37% and the least common was the luminal B and hybrid types each comprising 8%. The most common grade for HER2 was Grade III (50%). The association of histological grade with the molecular classication was statistically signicant with the p value of 0.01. Basal type (56%) had the highest incidence of N3 stage. ER, PR, HER2 neu, CK5/6 expression and proliferation index with Ki67 had a statistically signicant association with the molecular classication. High proliferation index (>14%) with Ki67 was noted in Luminal B, Basal and Hybrid types. 78% of the total 60 cases were alive and healthy. One death was reported in HER2, Hybrid and Basal types. The negative kappa value obtained while studying the agreement between the histopathological and molecular classication, indicates that the agreement is worse than chance and hence the importance of molecular classication is substantiated for the targeted therapy.

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Endoscopic Biopsy Pathology of Helicobacter pylori Gastritis

Archives of Pathology & Laboratory Medicine ◽

10.5858/1999-123-0778-ebpohp ◽

1999 ◽

Vol 123 (9) ◽

pp. 778-781 ◽

Cited By ~ 2

Author(s):

Maher Toulaymat ◽

Sharon Marconi ◽

Jane Garb ◽

Christopher Otis ◽

Shirin Nash

Keyword(s):

Helicobacter Pylori ◽

Specific Antibody ◽

Cost Effective ◽

Endoscopic Biopsy ◽

Immunohistochemical Method ◽

Labor Costs ◽

Chronic Active Gastritis ◽

Cost Effective Method ◽

H Pylori ◽

Helicobacter Pylori Gastritis

Abstract Objectives.—To describe the endoscopic biopsy pathology of Helicobacter pylori gastritis, compare bacterial detection by immunohistochemistry using a specific antibody with the Genta stain, and to compare the relative costs of the 2 techniques. Design.—One hundred cases of gastritis identified as positive for H pylori by Genta stain and 100 cases considered negative by the same technique were stained using an anti-H pylori–specific polyclonal antibody. Laboratory reagent and labor costs for the 2 methods were compared. Results.—Chronic active gastritis with lymphoid follicles was significantly associated with H pylori infection (P < .0001). The immunohistochemical method had a sensitivity of 97% and a specificity of 98% compared with the Genta stain, with strong agreement for grading density of organisms (κ = 0.85; P < .001). Reagent costs were similar for both methods, but immunohistochemistry using an autoimmunostainer required less dedicated technical time and hence was less expensive than the Genta stain. Conclusions.—Immunohistochemistry using a specific antibody is an accurate and cost-effective method for H pylori detection in gastric biopsies.

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Esophageal Small Cell Carcinomas

Archives of Pathology & Laboratory Medicine ◽

10.5858/2000-124-0228-escc ◽

2000 ◽

Vol 124 (2) ◽

pp. 228-233 ◽

Cited By ~ 3

Author(s):

King Y. Lam ◽

Simon Law ◽

Peter H. M. Tung ◽

John Wong

Keyword(s):

Cell Carcinoma ◽

Small Cell Carcinoma ◽

Survival Data ◽

Malignant Tumors ◽

P53 Protein ◽

Small Cell ◽

Immunohistochemical Method ◽

Clinicopathologic Features ◽

P53 Expression ◽

Mib 1

Abstract Objective.—To evaluate the clinicopathologic features and the roles of p53 and MIB-1 in esophageal small cell carcinoma. Method.—Twenty patients (14 men and 6 women) with esophageal small cell carcinoma treated in our hospital from 1982 through 1996 were studied. The clinicopathologic features, treatment received, and survival data of these patients were documented. Representative tissue was collected from each tumor, and immunohistochemical preparations for p53 protein and MIB-1 were made. Results.—Small cell carcinoma accounted for 1.3% of all esophageal malignant tumors. The median age of patients at presentation was 60 years. On gross examination, the tumors were large ulcerative lesions (median length, 7.5 cm). In 17 patients in whom p53 immunohistochemical study was performed, p53 protein was detected in 65% (9 of 17). All stage IV tumors were negative for p53 expression. The median tumor cell MIB-1 score was high at 855 (range, 810–964) positive cells per 1000. Overall median survival was 3.4 months. In patients who underwent chemotherapy, there was significant response. Conclusions.—Esophageal small cell carcinoma is an aggressive tumor. Overexpression of p53 is associated with early stages of carcinogenesis. The high proliferative index, as defined by the MIB-1 immunohistochemical method, may be related to aggressive behavior and high sensitivity to chemotherapy and radiotherapy.

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THE CAPILLARY PATTERN IN HUMAN MASSETER MUSCLE DURING AGEING

Image Analysis & Stereology ◽

10.5566/ias.v32.p135-144 ◽

2013 ◽

Vol 32 (3) ◽

pp. 135 ◽

Cited By ~ 1

Author(s):

Erika Cvetko ◽

Jiří Janáček ◽

Lucie Kubínová ◽

Ida Eržen

Keyword(s):

Muscle Fibre ◽

Masseter Muscle ◽

3D Visualization ◽

Fibre Length ◽

Fibre Surface ◽

Fibre Volume ◽

Immunohistochemical Method ◽

Muscle Fibres ◽

Age Related ◽

Novel Approach

The effect of ageing on the capillary network in skeletal muscles has produced conflicting results in both, human and animals studies. Some of the inconsistencies are due to non-comparable and biased methods that were applied on thin transversal sections, especially in muscles with complicated morphological structures, such as in human masseter muscle. We present a new immunohistochemical method for staining capillaries and muscle fibres in 100 µm thick sections as well as novel approach to 3D visualization of capillaries and muscle fibres. Applying confocal microscopy and virtual 3D stereological grids, or tracing capillaries in virtual reality, length of capillaries within a muscle volume or length of capillaries adjacent to muscle fibre per fibre length, fibre surface or fibre volume were evaluated in masseter muscle of young and old subjects by an unbiased approach. Our findings show that anatomic capillarity is well maintained in masseter muscle in old subjects; however, vascular remodelling occurs with age, which could be a response to changed muscle function and age-related muscle fibre type transformations.

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