scholarly journals Lactoferrin: nuclear localization in the human neutrophilic granulocyte?

1981 ◽  
Vol 29 (10) ◽  
pp. 1128-1136 ◽  
Author(s):  
R C Briggs ◽  
W F Glass ◽  
M M Montiel ◽  
L S Hnilica
Keyword(s):  
Human Peripheral Blood ◽  
Nuclear Materials ◽  
Major Protein ◽  
Staining Reaction ◽  
Nuclear Staining ◽  
Immunocytochemical Localization ◽  
Cytoplasmic Staining ◽  
Neutrophilic Granulocyte ◽  
Blood Smears ◽  
Blood Granulocyte

Conditions were established whereby nuclear or cytoplasmic immunocytochemical localization of lactoferrin was observed in the human peripheral blood granulocyte. A positive nuclear staining reaction was obtained when cells were either not fixed or treated with most fixatives. However, treatment of blood smears with formaldehydeacetone prior to the immunocytochemical localization gave a cytoplasmic staining reaction. A method was developed that allowed us to examine proteins obtained from granulocyte nuclei isolated from fixed cells. Only a trace amount of lactoferrin was detected in the electrophoretically separated nuclear proteins obtained from granulocytes treated for 1 min with formaldehyde-acetone. However, lactoferrin was major protein component in nuclei isolated from untreated cells, cells treated with other fixatives, or cells preincubated in buffered saline prior to formaldehyde-acetone fixation. formaldehyde-acetone treatment of nuclear materials containing lactoferrin did not extract lactoferrin or mask it from detection, thus indicating that lactoferrin found in the mature neutrophilic granulocyte nucleus is most likely acquired from other cellular organelles during tissue processing.

scholarly journals WT1 and Cyclin D1 Immunohistochemistry: A Useful Adjunct for Diagnosis of Pediatric Small Round Blue Cell Tumors on Small Biopsies

Diagnostics ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 2254
Author(s):  
Lucia Salvatorelli ◽  
Rosalba Parenti ◽  
Giuseppe Broggi ◽  
Giada Maria Vecchio ◽  
Giuseppe Angelico ◽  
...  
Keyword(s):  
Differential Diagnosis ◽  
Cyclin D1 ◽  
Wilms Tumor ◽  
Lymphoblastic Lymphoma ◽  
Routine Practice ◽  
Nuclear Staining ◽  
Cytoplasmic Staining ◽  
Blue Cell ◽  
Over Time

Pediatric small round blue cell tumors (SRBCTs) are a heterogeneous group of neoplasms with overlapping morphological appearance. Accordingly, their diagnosis is one of the most difficult in the field of surgical pathology. The most common tumors include rhabdomyosarcoma, Ewing’s sarcoma, neuroblastoma, lymphoblastic lymphoma and Wilms’ tumor (the blastemal component). Over time their diagnosis has become more difficult due to the increasing use of small biopsies. However, the advent of immunohistochemistry has improved the quality of diagnosis in most cases by the application of an adequate panel of immunomarkers. Recently, WT1 and Cyclin D1 have been shown to be useful in the differential diagnosis of SRBCTs on surgically-resected specimens, showing a diffuse cytoplasmic positivity of the former in all RMSs and a diffuse nuclear staining of the latter in both EWS and NB. The aim of the present study was to investigate the expression of WT1 and Cyclin D1 on small biopsies from a series of 105 pediatric SRBCTs to evaluate their diagnostic utility. Both immunomarkers were differentially expressed, with a diffuse and strong cytoplasmic staining for WT1 limited to all cases of RMS, and a diffuse nuclear staining for cyclin D1 restricted to all cases of EWS and NB. Notably, the expression of WT1 and cyclin D1 was also retained in those cases in which the conventional tumor markers (myogenin, desmin and MyoD1 for RMS; CD99 for EWS; NB84 for NB) were focally expressed or more rarely absent. The present study shows that WT1 and Cyclin D1 are helpful immunomarkers exploitable in the differential diagnosis of pediatric SRBCTs on small biopsies, suggesting their applicability in routine practice.

scholarly journals A substitute to xylene in deparaffinization and clearing prior to coverslipping in histopathology

2019 ◽  
Vol 11 (02) ◽  
pp. 118-122
Author(s):  
Nasar Yousuf Alwahaibi ◽  
Sirin Hamed Aldughaishi
Keyword(s):  
Cost Benefit ◽  
Cost Effective ◽  
Nuclear Staining ◽  
Laboratory Cost ◽  
Cytoplasmic Staining ◽  
Group A ◽  
Staining Methods ◽  
Laboratory Workers ◽  
Group B ◽  
Group D

Abstract INTRODUCTION: Deparaffinization and clearing prior to coverslipping are important steps in all staining methods in histopathology. Xylene is the most commonly used agent worldwide. However, xylene is toxic. We evaluated safer alternative dewaxing and clearing agents prior to coverslipping in a histopathology laboratory. MATERIALS AND METHODS: Thirteen different fresh surgical tissues were cut into two halves. One half processed using xylene and the other half processed using UltraClear™. Five groups were designed. For each Group of A, B, C, and D, 100 slides were cut from xylene-processed blocks. For Group E, 100 slides were cut from UltraClear™-processed blocks. Group A is the standard method. Group B evaluates UltraClear™ as a dewaxing agent only. Group C evaluates UltraClear™ as a clearing agent prior to coverslipping only. Group D evaluates UltraClear™ as both dewaxing and clearing agents prior to coverslipping. Group E evaluates UltraClear™ as both dewaxing and clearing agents prior to coverslipping. Six parameters were evaluated: nuclear staining, cytoplasmic staining, cell morphology, clarity of staining, uniformity of staining, and cost. RESULTS: Groups B, C, and D showed 79% (P = 0.054), 83% (P = 0.221), and 80% (P = 0.079) adequacy when compared with Group A (89%), respectively. However, Group E showed only 76% (P = 0.016) adequacy. UltraClear™ is more expensive than xylene. CONCLUSION: UltraClear™ is a promising dewaxing agent. It is also a good clearing agent for use prior to coverslipping in histopathology laboratory. Cost-benefit balance between safety of laboratory workers, good quality staining, and cost-effective strategy needs to be further studied.

scholarly journals β-catenin activation is not involved in sporadic parathyroid carcinomas and adenomas

2010 ◽  
Vol 17 (1) ◽  
pp. 1-6 ◽  
Author(s):  
F Cetani ◽  
E Pardi ◽  
C Banti ◽  
P Collecchi ◽  
P Viacava ◽  
...  
Keyword(s):  
Cell Membrane ◽  
Signaling Pathway ◽  
Direct Sequencing ◽  
Distant Metastases ◽  
Outer Cell ◽  
Tumor Type ◽  
Nuclear Staining ◽  
Cytoplasmic Staining ◽  
Membrane Staining ◽  
Outer Cell Membrane

Aberrant accumulation of β-catenin has been found in various types of human tumors. The aim of this study was to evaluate whether Wnt/β-catenin signaling is activated in parathyroid carcinomas and adenomas. We studied 154 parathyroid tumors (18 carcinomas (13 with distant metastases), six atypical adenomas, and 130 adenomas). Three normal parathyroid tissues were used as control. Direct sequencing of exon 3 of the CTNNB1 gene showed absence of stabilizing mutations in all the tumors. Immunostaining of β-catenin was performed in all carcinomas and in 66 adenomas (including three atypical). Normal parathyroid showed a homogeneous distinct outer cell membrane staining in the majority of cells and no nuclear staining. A weak cytoplasmic staining was observed in one case. All tumors showed negative nuclear staining. With the exception of one carcinoma, which had a negative membrane staining, all other samples showed a membrane staining which was similar to that of the normal parathyroid. β-Catenin expression was heterogeneous with a range of positive cells between 5 and 80%, independently of tumor type. Our results suggest that the Wnt/β-catenin signaling pathway is not involved in the development of parathyroid carcinomas and adenomas.

scholarly journals Effects of Estradiol on Prostate Epithelial Cells in the Castrated Rat

2002 ◽  
Vol 50 (11) ◽  
pp. 1517-1523 ◽  
Author(s):  
G. Pelletier
Keyword(s):  
Epithelial Cells ◽  
Cdna Probe ◽  
Concomitant Administration ◽  
Hybridization Signal ◽  
Mrna Levels ◽  
Nuclear Staining ◽  
Cytoplasmic Staining ◽  
Prostate Epithelial Cells ◽  
Strong Hybridization ◽  
Rat Prostate

There is evidence that estrogens can modulate the activity of prostate epithelial cells. To determine whether estradiol can have a direct influence on rat prostate, this study examined the effects of estradiol-17β (E2) administered alone or in combination with dihydrotestosterone (DHT) to castrated rats for 3 weeks on prostate binding protein (PBP) C1 mRNA expression and androgen receptor (AR) localization. PBP C1 mRNA levels were measured by semi-quantitative in situ hybridization using a 35S-labeled cDNA probe. In intact animals, strong hybridization signal could be observed in prostate sections after 12 hr of exposure to Kodak X-Omat films. In castrated rats, no PBP C1 mRNA could be detected even with longer exposure times, an effect that was prevented by administration of DHT. E2 administered alone induced a detectable hybridization signal, and the concomitant administration of E2 and DHT induced an increase in PBP C1 mRNA that significantly exceeded that obtained in animals that received only DHT. In prostate epithelial cells of intact animals, AR immunostaining was restricted to the nucleus. In castrated animals the alveoli were decreased in size and the epithelial cells were atrophied. AR staining was weak and was detected in both cytoplasm and nucleus. DHT administration completely obviated the effect of castration on epithelial cell histology and on AR immunostaining distribution and intensity. Interestingly, E2 administration alone induced moderate hypertrophy of epithelial cells compared to the histological appearance of cells in untreated castrated rats. Moreover, in E2-treated animals the nuclear staining was much stronger than that detected in untreated castrated rats, whereas the cytoplasmic staining was not modified by the treatment. In animals that received both DHT and E2, the staining was similar to that seen in DHT-treated rats. These results suggest that E2 can influence the activity of rat prostate epithelial cells by mechanisms that remain to be fully clarified.

scholarly journals Mechanistic Study of Triazole Based Aminodiol Derivatives in Leukemic Cells—Crosstalk between Mitochondrial Stress-Involved Apoptosis and Autophagy

2020 ◽  
Vol 21 (7) ◽  
pp. 2470
Author(s):  
She-Hung Chan ◽  
Wohn-Jenn Leu ◽  
Sharada Prasanna Swain ◽  
Jui-Ling Hsu ◽  
Duen-Ren Hou ◽  
...  
Keyword(s):  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Cholesterol Biosynthesis ◽  
Leukemic Cell ◽  
Leukemic Cells ◽  
Mechanistic Study ◽  
Nuclear Staining ◽  
Peripheral Blood Mononuclear ◽  
Mitochondrial Stress ◽  
Stat3 Phosphorylation

Various derivatives that mimic ceramide structures by introducing a triazole to connect the aminodiol moiety and long alkyl chain have been synthesized and screened for their anti-leukemia activity. SPS8 stood out among the derivatives, showing cytotoxic selectivity between leukemic cell lines and human peripheral blood mononuclear cells (about ten times). DAPI nuclear staining and H&E staining revealed DNA fragmentation under the action of SPS8. SPS8 induced an increase in intracellular Ca2+ levels and mitochondrial stress in HL-60 cells identified by the loss of mitochondrial membrane potential, transmission electron microscopy (TEM) examination, and altered expressions of Bcl-2 family proteins. SPS8 also induced autophagy through the detection of Atg5, beclin-1, and LC3 II protein expression, as well as TEM examination. Chloroquine, an autophagy inhibitor, promoted SPS8-induced apoptosis, suggesting the cytoprotective role of autophagy in hindering SPS8 from apoptosis. Furthermore, SPS8 was shown to alter the expressions of a variety of genes using a microarray analysis and volcano plot filtering. A further cellular signaling pathways analysis suggested that SPS8 induced several cellular processes in HL-60, including the sterol biosynthesis process and cholesterol biosynthesis process, and inhibited some cellular pathways, in which STAT3 was the most critical nuclear factor. Further identification revealed that SPS8 inhibited the phosphorylation of STAT3, representing the loss of cytoprotective activity. In conclusion, the data suggest that SPS8 induces both apoptosis and autophagy in leukemic cells, in which autophagy plays a cytoprotective role in impeding apoptosis. Moreover, the inhibition of STAT3 phosphorylation may support SPS8-induced anti-leukemic activity.

scholarly journals THE DISTRIBUTION OF HISTOCHEMICALLY DEMONSTRABLE GLYCOGEN IN HUMAN BLOOD AND BONE MARROW CELLS

Blood ◽  
1949 ◽  
Vol 4 (1) ◽  
pp. 54-59 ◽  
Author(s):  
MAX WACHSTEIN
Keyword(s):  
Bone Marrow ◽  
Human Blood ◽  
Acid Treatment ◽  
Bone Marrow Cells ◽  
Periodic Acid ◽  
Myeloid Cells ◽  
Staining Reaction ◽  
Cytoplasmic Staining ◽  
Chemical Association ◽  
Marrow Cells

Abstract By applying Schiff’s reagent after periodic acid treatment to blood and bone marrow films, a cytoplasmic staining reaction is seen in some cells of the myeloid series, as well as in megakaryocytes and platelets. The intensity of the staining reaction in the myeloid cells increases with their maturation. The staining reaction can be prevented altogether in alcohol-fixed films by salivary digestion, but only incompletely in air-dried films. The staining reaction is due to the presence of glycogen in some chemical association, possibly with protein.

Association of DNA repair factors with overall survival in advanced urothelial carcinoma treated with platinum-based chemotherapy.

2012 ◽  
Vol 30 (5_suppl) ◽  
pp. 291-291
Author(s):  
Elizabeth Ann Guancial ◽  
Lillian Werner ◽  
Edward Stack ◽  
Rosina Lis ◽  
Sabina Signoretti ◽  
...  
Keyword(s):  
Dna Damage ◽  
Overall Survival ◽  
Dna Repair ◽  
Urothelial Carcinoma ◽  
Metastatic Disease ◽  
Tumor Tissue ◽  
Cytotoxic Agents ◽  
Nuclear Staining ◽  
Prognostic Variables ◽  
Cytoplasmic Staining

291 Background: DNA repair factors are hypothesized to mediate chemosensitivity to cytotoxic agents that produce DNA damage and may be predictive for response to platinum-based chemotherapeutic regimens. Primary urothelial carcinoma (UC) tumors of patients who developed metastatic disease were evaluated for expression of a panel of DNA repair factors by immunohistochemistry (IHC). Methods: A cohort of 132 clinically annotated, uniformly-treated (platinum-based combination chemotherapy) UC patients who subsequently developed distant metastases with FFPE primary tumor tissue available was identified. Tumor bearing areas were evaluated by a single urologic pathologist. Tissue arrays were constructed for IHC analysis of the following DNA repair factors: ERCC1, Rad 51, BRCA1/2, PAR and PARP1. Tumor was deparaffinized and specific antigen retrieval determined for individual antibodies. Pathologist supervised IHC analysis of nuclear versus cytoplasmic expression was performed using spectral imaging analysis. Overall survival (OS) was defined from start of chemotherapy for metastatic disease to death (N=67) or censored on the last known alive date. Percent of positive nuclear staining was categorized as quartiles or previously identified cut-points. Cox regression evaluated the associations of percent positive nuclear staining levels and OS in multivariable analysis (HRs and 95% CIs included) that controlled for known prognostic variables (performance status and visceral metastases). Results: Higher percentage of nuclear staining of ERCC1 [HR=2.7 (1.5, 4.9), p=0.0007]; Rad51 [HR=5.6 (1.7, 18.3), p=0.005]; and PAR [HR=2.2 (1.1, 4.4), p=0.026] in tumor tissue were associated with poor outcome; each was independent of the other two repair factors and known prognostic variables. Neither nuclear nor cytoplasmic staining for BRCA1/2 or PARP1 reached statistical significance for an association with OS. Conclusions: DNA damage repair factor levels measured by IHC impact outcomes in advanced UC. External validation is ongoing and will be presented. Further studies are required to determine whether these biomarkers are prognostic or predictive in this setting.

FBI-1 expression in relation to clinicopathological variables in rectal cancers with a Swedish clinical trial of preoperative radiotherapy.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e15102-e15102
Author(s):  
Chao-Jie Wang ◽  
Jian-Wei Zhou ◽  
Yun Zhou ◽  
Xiao-Feng Sun
Keyword(s):  
Clinical Trial ◽  
Rectal Cancer ◽  
Preoperative Radiotherapy ◽  
Disease Free Survival ◽  
Normal Mucosa ◽  
Tnm Stage ◽  
Nuclear Staining ◽  
Cytoplasmic Staining ◽  
Rectal Cancers ◽  
Pattern Differentiation

e15102 Background:FBI-1 is a recently characterized proto-oncoprotein of the POZ domain Krüppel-like (POK) family of transcription factors. Although several studies provide the evidence that FBI-1 is an important gene regulator in CRC, no analysis of any correlation between FBI-1 expression and preoperative radiotherapy (RT) has been studied in rectal cancers. Methods: This study included the patients with rectal cancer that participated in a Swedish clinical trial of preoperative RT between 1987 and 1990. Patients were divided into preoperative RT (62) and non-RT (77) groups. Applying immunohistochemstry, we detected FBI-1 expression in 118 normal mucosa, 139 primary rectal cancers, and 45 lymph node metastases, and analyzed its relationship with clinicopathological features and RT response. Results: FBI-1 was detected both in the cytoplasm and nucleus, and the cytoplasmic staining was up-regulated compared with normal mucosa both in non-RT and RT groups (74.0% vs. 17.7%, p < 0.001; 69.4% vs. 41.1%, p = 0.002), however, the nuclear staining was down-regulated compared with normal mucosa both in non-RT and RT groups (22.1% vs. 75.8%, p < 0.001; 35.5% vs. 83.9% p < 0.001). Both cytoplasmic and nuclear staining were no difference between the non-RT and RT groups (74.0% vs. 69.4%, p = 0.542; 22.1% vs. 35.5%, p = 0.080, respectively). So we combined the non-RT and RT group together for further analysis. Nuclear staining of FBI-1 was positively with TNM stage and distance recurrence, it showed higher expression in III+ IV stage than that in I+II stage (41.0% vs. 17.9%, p = 0.003). The patients with distance recurrence showed higher FBI-1 expression than that with no distance recurrence (39.7% vs. 19.8%, p = 0.010). In stage I, II and III patients, higher nuclear FBI-1 in primary cancer showed worse disease free survival (HR: 1.934; 95%CI: 1.055-3.579, p = 0.033) and overall survival (HR: 2.174; 95%CI: 1.102-4.290; p = 0.025) independent of gender, age, growth pattern, differentiation and RT. Conclusions: Higher nuclear FBI-1 is related with later TNM stage, distance recurrence, and worse prognosis, it can be used as a potential diagnostic and prognostic biomarker in rectal cancer.

Expression of antiapoptotic protein survivin in malignant melanoma

Biologia ◽  
2009 ◽  
Vol 64 (4) ◽  
Author(s):  
Marian Adamkov ◽  
L’udovít Lauko ◽  
Július Rajčáni ◽  
Soňa Bálentová ◽  
Silvia Rybárová ◽  
...  
Keyword(s):  
Malignant Melanoma ◽  
Tumor Cells ◽  
Immunohistochemical Staining ◽  
Diagnostic Marker ◽  
Nuclear Staining ◽  
Cytoplasmic Localization ◽  
Cytoplasmic Staining ◽  
Antiapoptotic Protein ◽  
Histological Features ◽  
Degree Of Malignancy

AbstractWe examined the expression of potential tumor marker survivin by immunohistochemical staining using antisurvivin antibody (DAKO, Clone 12C4) in a panel of 25 malignant melanomas. In each section, we assessed the percentage of positively stained tumor cells, the intensity of staining and its subcellular localization. Survivin was present in 23 out of 25 cases (92%). Nuclear staining was found in 2 of these 23 cases (8.7%) only, while cytoplasmic staining only was seen in 3 of them (13%). The combined nuclear as well as cytoplasmic localization of survivin was demonstrated in 18 out of 23 cases (78.3%). In 2 cases revealing nuclear staining only, the worse histological features were more pronounced than in 3 cases with cytoplasmic staining only. Our results suggest that nuclear positivity of survivin may correlate with the degree of malignancy. In addition, we conclude that overexpression of survivin involved in the pathogenesis of melanoma represents an important diagnostic marker.

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