A correlative chemical and histochemical study of the O-acetylated sialic acids of human colonic epithelial glycoproteins in formalin fixed paraffin embedded tissues.

Procedures are described for the isolation and identification of epithelial glycoproteins from formalin fixed, paraffin embedded specimens of human large intestine. The side chain O-acetylation patterns of the sialic acids of these glycoproteins were surprisingly similar to those of purified glycoproteins prepared from epithelial cells obtained from the same tissue before fixation. These results were consistent with those obtained by histochemical procedures performed on representative sections taken from the same tissue blocks. The methodology described permits a direct correlation of chemical and histochemical results obtained from the study of colonic epithelial glycoproteins of both normal and diseased tissues. It eliminates some of the difficulties associated with interpretation of the results by either discipline and may provide new information which would be unavailable by either chemistry or histochemistry alone.

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Transcriptome analyses of benign and malignant prostate epithelial cells in formalin-fixed paraffin-embedded whole-mounted radical prostatectomy specimens

Prostate Cancer and Prostatic Diseases ◽

10.1038/sj.pcan.4501007 ◽

2007 ◽

Vol 11 (2) ◽

pp. 194-197 ◽

Cited By ~ 22

Author(s):

B Furusato ◽

S Shaheduzzaman ◽

G Petrovics ◽

A Dobi ◽

M Seifert ◽

...

Keyword(s):

Radical Prostatectomy ◽

Epithelial Cells ◽

Prostate Epithelial Cells ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Transcriptome Analyses ◽

Malignant Prostate ◽

Formalin Fixed

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Detection of Mycoplasma ovipneumoniae and Pasteurella haemolytica Antigens by an Immunoperoxidase Technique in Pneumonic Ovine Lungs

Veterinary Pathology ◽

10.1177/030098589603300108 ◽

1996 ◽

Vol 33 (1) ◽

pp. 74-76 ◽

Cited By ~ 9

Author(s):

R. Haziroglu ◽

K. S. Diker ◽

J. Turkarslan ◽

M. Y. Gulbahar

Keyword(s):

Epithelial Cells ◽

Pasteurella Haemolytica ◽

Histologic Examination ◽

Complex Method ◽

Immunoperoxidase Technique ◽

Tissue Sections ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Formalin Fixed ◽

Mycoplasma Ovipneumoniae

Four hundred twenty pneumonic lungs from lambs were examined for Mycoplasma ovipneumoniae and Pasteurella haemolytica by an immunoperoxidase technique using an extravidin-biotin-peroxidase complex method in formalin-fixed, paraffin-embedded sections. Histologic examination of tissue sections revealed strong positive reactions in 60.9% and 68.3% of the lungs against M. ovipneumoniae and P. haemolytica, respectively. M. ovipneumoniae and P. haemolytica antigens were observed at the surface and/or within the epithelial cells, macrophages, leucocytes, and bronchiolar exudate. The location of M. ovipneumoniae in the cytoplasm of the epithelial cells and P. haemolytica in the neutrophils was detected immunohistochemically.

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Distribution of Tight Junction Proteins in Adult Human Salivary Glands

Journal of Histochemistry & Cytochemistry ◽

10.1369/jhc.2008.951780 ◽

2008 ◽

Vol 56 (12) ◽

pp. 1093-1098 ◽

Cited By ~ 30

Author(s):

Ola M. Maria ◽

Jung-Wan Martin Kim ◽

Jonathan A. Gerstenhaber ◽

Bruce J. Baum ◽

Simon D. Tran

Keyword(s):

Endothelial Cells ◽

Salivary Gland ◽

Salivary Glands ◽

Adult Human ◽

New Information ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Molecular Components ◽

Formalin Fixed ◽

Junction Proteins

Tight junctions (TJs) are an essential structure of fluid-secreting cells, such as those in salivary glands. Three major families of integral membrane proteins have been identified as components of the TJ: claudins, occludin, and junctional adhesion molecules (JAMs), plus the cytosolic protein zonula occludens (ZO). We have been working to develop an orally implantable artificial salivary gland that would be suitable for treating patients lacking salivary parenchymal tissue. To date, little is known about the distribution of TJ proteins in adult human salivary cells and thus what key molecular components might be desirable for the cellular component of an artificial salivary gland device. Therefore, the aim of this study was to determine the distribution of TJ proteins in human salivary glands. Salivary gland samples were obtained from 10 patients. Frozen and formalin-fixed paraffin-embedded sections were stained using IHC methods. Claudin-1 was expressed in ductal, endothelial, and ∼25% of serous cells. Claudins-2, −3, and −4 and JAM-A were expressed in both ductal and acinar cells, whereas claudin-5 was expressed only in endothelial cells. Occludin and ZO-1 were expressed in acinar, ductal, and endothelial cells. These results provide new information on TJ proteins in two major human salivary glands and should serve as a reference for future studies to assess the presence of appropriate TJ proteins in a tissue-engineered human salivary gland.

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Immunohistochemical Detection of Hog Cholera Viral Glycoprotein 55 in Paraffin-Embedded Tissues

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063879700900103 ◽

1997 ◽

Vol 9 (1) ◽

pp. 10-16 ◽

Cited By ~ 25

Author(s):

Juana Martín de las Mulas ◽

Eduardo Ruiz-Villamor ◽

Sergio Donoso ◽

Manuel Quezada ◽

Claudio Lecocq ◽

...

Keyword(s):

Epithelial Cells ◽

Lymphoid Cells ◽

Esophageal Mucosa ◽

Viral Glycoprotein ◽

Diarrhea Virus ◽

Formalin Fixed Paraffin ◽

Hog Cholera ◽

Hog Cholera Virus ◽

Formalin Fixed Paraffin Embedded ◽

Formalin Fixed

Formalin-fixed, paraffin embedded tissues obtained from 40 pigs inoculated with a field isolate of hog cholera virus were examined for the presence of Gp55, a major structural protein of the virus envelope, using a monoclonal antibody-based immunohistochemical test with the avidin-biotin-peroxidase complex method. Immunoreactivity was detected in hog cholera virus-infected tissues but not in control pigs tissues, African swine fever virus-infected tissues, or bovine viral diarrhea virus-infected porcine or bovine tissues. The first positive reactions were seen in lymphatic tissues, digestive tract and skin on postinoculation day (pid) 4, respiratory and urinary tissues on pid 5, nervous tissues on pid 6, and endocrine tissues on pid 7. These staining reactions persisted until the last observation on pid 18. Hog cholera virus antigen was not detected in heart tissue at any time. The highest levels of antigen detection were found in tonsils, spleen, and pancreas, although the esophageal mucosa and skin epithelial cells were also intensely and widely stained. The cellular staining pattern of Gp55 had a ubiquitous distribution. It was found in epithelial cells, macrophages and circulating monocytes, endothelial cells, lymphoid cells, and glial cells. The results showed a high specificity and high sensitivity for detecting hog cholera Gp55 in formalin-fixed, paraffin embedded tissue samples. This method allows precise association of Gp55 with specific cells, tissues, and histologic lesions, making the technique suitable for use in routine diagnosis of hog cholera.

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Nested Polymerase Chain Reaction and In Situ Hybridization for Diagnosis of Canine Herpesvirus Infection in Puppies

Veterinary Pathology ◽

10.1177/030098589803500306 ◽

1998 ◽

Vol 35 (3) ◽

pp. 209-217 ◽

Cited By ~ 38

Author(s):

C. Schulze ◽

W. Baumgärtner

Keyword(s):

Epithelial Cells ◽

Chain Reaction ◽

Viral Dna ◽

Nested Polymerase Chain Reaction ◽

Canine Herpesvirus ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Polymerase Chain ◽

Formalin Fixed

The usefulness of two nucleic acid detection systems in suspected cases of spontaneous canine herpesvirus (CHV) infection in puppies was evaluated. Formalin-fixed, paraffin-embedded tissues from seven 1–3-week-old naturally infected puppies with lesions characteristic of CHV infection were investigated in a retrospective study. Nested polymerase chain reaction (PCR) and nonradioactive in situ hybridization (ISH) were used to detect nucleotide sequences of the CHV thymidine kinase (TK) gene. According to the original necropsy reports, CHV was isolated in four of the seven puppies using primary canine lung and/or kidney cells. In all seven puppies, gross and histologic lesions consisted of disseminated focal necroses and hemorrhages predominantly in kidneys, lung, liver, and spleen. In addition, few small amphophilic intranuclear inclusion bodies were detected by light microscopy mainly in epithelial cells of kidney, lung, and liver. ISH was performed with a 111-base-pair (bp) digoxigenin-labeled double-stranded DNA probe. Viral DNA was detected in the nuclei of cells near and within lesions. Various cell types, including bronchiolar and alveolar epithelial cells, hepatocytes, renal tubular epithelial cells, neurons, fibrocytes, cardiac myocytes, and endothelial cells, were positive for viral DNA. PCR amplification products of the expected length of 168 bp containing the expected cleavage site for the restriction enzyme EcoRI, derived from paraffin blocks containing lung, kidney, and liver tissues, were detected in all seven puppies. The specificity of the obtained amplicon was further confirmed by Southern blot analysis. ISH and PCR are both useful methods for diagnosing CHV infection in formalin-fixed, paraffin-embedded tissues and are highly specific and sensitive methods for further investigations of the pathogenesis of CHV-induced lesions.

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218.FGF9 stimulates ovarian progesterone production

Reproduction Fertility and Development ◽

10.1071/srb04abs218 ◽

2004 ◽

Vol 16 (9) ◽

pp. 218

Author(s):

A. E. Drummond ◽

M. Dyson ◽

J. K. Findlay

Keyword(s):

Granulosa Cells ◽

Side Chain ◽

Immature Rat ◽

Progesterone Production ◽

Chain Cleavage ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Rat Ovary ◽

The Impact ◽

Formalin Fixed

FGF9, a member of the fibroblast growth factor family (FGF), is known to be a male sex-determining factor involved in testicular cord formation (1). FGF9 knockout males are sex-reversed (2). However, nothing is known about FGF9's role in folliculogenesis because these mice die at birth (2). We previously reported the presence of FGF9 mRNA and protein in the immature rat ovary (3). In these studies we investigated: (1) the presence of FGF9 receptors (FGFR3) on granulosa cells (GC); and (2) the impact of FGF9 on GC progesterone production. GC isolated from 21 day old diethylstilboestrol (DES)-treated rats were cultured for either 2 hours (RNA) or 2 days (progesterone) in McCoys 5C with FGF9 (0.1-50ng/ml) � FSH (100ng/ml). Progesterone was measured in conditioned media by radioimmunoassay. RNA was extracted from the granulosa cells and reverse-transcribed for PCR. Specific primers for P450 side chain cleavage (SCC) amplified a 329�bp cDNA fragment. GAPDH was used for data normalisation. The FGF9 receptor FGFR3, was immunolocalised on formalin-fixed, paraffin-embedded sections of immature rat ovary. FGFR3 protein was localised only to GC of the ovary. Progesterone production by cultured GC was significantly elevated by FGF9, consistent with the presence of FGFR3. Relative to a maximally stimulating dose of FSH, FGF9 increased progesterone production 10- fold. In preliminary studies, FGF9 increased the expression of P450 SCC mRNA by cultured GC revealing a mechanism by which FGF9 increases progesterone production. These data suggest a role for FGF9 not just in testicular formation, but in the regulation of ovarian steroidogenesis. Supported by the NH&MRC of Australia (Regkey 241000 & 198705). (1) Cotinot et al. (2002) Semin. Reprod. Med. 20, 157. (2) Colvin et al. (2001) Cell 104, 875. (3) Drummond et al. (2003) SRB Abstract 90.

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