Immunohistochemical Analyses on Albumin and IgG in Acute Hypertensive Mouse Kidneys

Background: Cadmium (Cd) impairs gametogenesis and damages the blood-testis barrier. Objective: As the primary mechanism of Cd-induced damage is oxidative stress, the effects of two natural antioxidants, myo-inositol (MI) and seleno-L-methionine (Se), were evaluated in mice testes. Methods: Eighty-four male C57 BL/6J mice were divided into twelve groups: 0.9% NaCl (vehicle; 1 ml/kg/day i.p.); Se (0.2 mg/kg/day per os); Se (0.4 mg/kg/day per os); MI (360 mg/kg/day per os); MI plus Se (0.2 mg/kg/day); MI plus Se (0.4 mg/kg/day); CdCl2 (2 mg/kg/day i.p.) plus vehicle; CdCl2 plus MI; CdCl2 plus Se (0.2 mg/kg/day); CdCl2 plus Se (0.4 mg/kg/day); CdCl2 plus MI plus Se (0.2 mg/kg/day); and CdCl2 plus MI plus Se (0.4 mg/kg/day). After 14 days, testes were processed for biochemical, structural and immunohistochemical analyses. Results: CdCl2 increased iNOS and TNF-α expression and Malondialdehyde (MDA) levels, lowered glutathione (GSH) and testosterone, induced testicular lesions, and almost eliminated claudin-11 immunoreactivity. Se administration at 0.2 or 0.4 mg/kg significantly reduced iNOS and TNF-α expression, maintained GSH, MDA and testosterone levels, structural changes and low claudin-11 immunoreactivity. MI alone or associated with Se at 0.2 or 0.4 mg/kg significantly reduced iNOS and TNF-α expression and MDA levels, increased GSH and testosterone levels, ameliorated structural organization and increased claudin-11 patches number. Conclusion: We demonstrated a protective effect of MI, a minor role of Se and an evident positive role of the association between MI and Se on Cd-induced damages of the testis. MI alone or associated with Se might protect testes in subjects exposed to toxicants, at least to those with behavior similar to Cd.

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Gene expression and immunohistochemical analyses identify SOX2 as major risk factor for overall survival and relapse in Ewing sarcoma patients

EBioMedicine ◽

10.1016/j.ebiom.2019.08.002 ◽

2019 ◽

Vol 47 ◽

pp. 156-162 ◽

Cited By ~ 5

Author(s):

Giuseppina Sannino ◽

Aruna Marchetto ◽

Andreas Ranft ◽

Susanne Jabar ◽

Constanze Zacherl ◽

...

Keyword(s):

Gene Expression ◽

Overall Survival ◽

Risk Factor ◽

Ewing Sarcoma ◽

Major Risk Factor ◽

Immunohistochemical Analyses

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Transglutaminases Are Active in Perivascular Adipose Tissue

International Journal of Molecular Sciences ◽

10.3390/ijms22052649 ◽

2021 ◽

Vol 22 (5) ◽

pp. 2649

Author(s):

Alexis N. Orr ◽

Janice M. Thompson ◽

Janae M. Lyttle ◽

Stephanie W. Watts

Keyword(s):

Adipose Tissue ◽

Vascular Remodeling ◽

Coagulation Factor ◽

Sprague Dawley ◽

Rt Pcr ◽

Ideal Model ◽

Perivascular Adipose Tissue ◽

Tissue Sections ◽

Insight Into ◽

Immunohistochemical Analyses

Transglutaminases (TGs) are crosslinking enzymes best known for their vascular remodeling in hypertension. They require calcium to form an isopeptide bond, connecting a glutamine to a protein bound lysine residue or a free amine donor such as norepinephrine (NE) or serotonin (5-HT). We discovered that perivascular adipose tissue (PVAT) contains significant amounts of these amines, making PVAT an ideal model to test interactions of amines and TGs. We hypothesized that transglutaminases are active in PVAT. Real time RT-PCR determined that Sprague Dawley rat aortic, superior mesenteric artery (SMA), and mesenteric resistance vessel (MR) PVATs express TG2 and blood coagulation Factor-XIII (FXIII) mRNA. Consistent with this, immunohistochemical analyses support that these PVATs all express TG2 and FXIII protein. The activity of TG2 and FXIII was investigated in tissue sections using substrate peptides that label active TGs when in a catalyzing calcium solution. Both TG2 and FXIII were active in rat aortic PVAT, SMAPVAT, and MRPVAT. Western blot analysis determined that the known TG inhibitor cystamine reduced incorporation of experimentally added amine donor 5-(biotinamido)pentylamine (BAP) into MRPVAT. Finally, experimentally added NE competitively inhibited incorporation of BAP into MRPVAT adipocytes. Further studies to determine the identity of amidated proteins will give insight into how these enzymes contribute to functions of PVAT and, ultimately, blood pressure.

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Simultaneous Expression of Th1- and Treg-Associated Chemokine Genes and CD4+, CD8+, and Foxp3+ Cells in the Premalignant Lesions of 4NQO-Induced Mouse Tongue Tumorigenesis

Cancers ◽

10.3390/cancers13081835 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1835

Author(s):

Hana Yamaguchi ◽

Miki Hiroi ◽

Kazumasa Mori ◽

Ryosuke Ushio ◽

Ari Matsumoto ◽

...

Keyword(s):

Immune Cell ◽

Tongue Cancer ◽

Immunohistochemical Analysis ◽

Premalignant Lesions ◽

Cytokine Gene Expression ◽

Cell Infiltration ◽

Mrna Levels ◽

Cell Recruitment ◽

T Cell Infiltration ◽

Immunohistochemical Analyses

Chemokines and cytokines in the tumor microenvironment influence immune cell infiltration and activation. To elucidate their role in immune cell recruitment during oral cancer development, we generated a mouse tongue cancer model using the carcinogen 4-nitroquinoline 1-oxide (4NQO) and investigated the carcinogenetic process and chemokine/cytokine gene expression kinetics in the mouse tongue. C57/BL6 mice were administered 4NQO in drinking water, after which tongues were dissected at 16 and 28 weeks and subjected to analysis using the RT2 Profiler PCR Array, qRT-PCR, and pathologic and immunohistochemical analyses. We found that Th1-associated chemokine/cytokine (Cxcl9, Cxcl10, Ccl5, and Ifng) and Treg-associated chemokine/cytokine (Ccl17, Ccl22, and Il10) mRNA levels were simultaneously increased in premalignant lesions of 4NQO-treated mice at 16 weeks. Additionally, although levels of Gata3, a Th2 marker, were not upregulated, those of Cxcr3, Ccr4, and Foxp3 were upregulated in the tongue tissue. Furthermore, immunohistochemical analysis confirmed the infiltration of CD4+, CD8+, and Foxp3+ cells in the tongue tissue of 4NQO-treated mice, as well as significant correlations between Th1- or Treg-associated chemokine/cytokine mRNA expression and T cell infiltration. These results indicate that CD4+, CD8+, and Foxp3+ cells were simultaneously recruited through the expression of Th1- and Treg-associated chemokines in premalignant lesions of 4NQO-induced mouse tongue tissue.

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Alteration of Tissue Marking Dyes Depends on Used Chromogen during Immunohistochemistry

Cells ◽

10.3390/cells10040835 ◽

2021 ◽

Vol 10 (4) ◽

pp. 835

Author(s):

Selina Kiefer ◽

Julia Huber ◽

Hannah Füllgraf ◽

Kristin Sörensen ◽

Agnes Csanadi ◽

...

Keyword(s):

Colour Change ◽

Residual Tumour ◽

Blue Colour ◽

Close Margin ◽

Correct Orientation ◽

Therapeutic Regime ◽

Colour Changes ◽

Tissue Specimens ◽

Immunohistochemical Analyses ◽

Colour Signal

Pathological biopsy protocols require tissue marking dye (TMD) for orientation. In some cases (e.g., close margin), additional immunohistochemical analyses can be necessary. Therefore, the correlation between the applied TMD during macroscopy and the examined TMD during microscopy is crucial for the correct orientation, the residual tumour status and the subsequent therapeutic regime. In this context, our group observed colour changes during routine immunohistochemistry. Tissue specimens were marked with various TMD and processed by two different methods. TMD (blue, red, black, yellow and green) obtained from three different providers (A, B and C, and Whiteout/Tipp-Ex®) were used. Immunohistochemistry was performed manually via stepwise omission of reagents to identify the colour changing mechanism. Blue colour from provider A changed during immunohistochemistry into black, when 3,3′-Diaminobenzidine-tetrahydrochloride-dihydrate (DAB) and H2O2 was applied as an immunoperoxidase-based terminal colour signal. No other applied reagents, nor tissue texture or processing showed any influence on the colour. The remaining colours from provider A and the other colours did not show any changes during immunohistochemistry. Our results demonstrate an interesting and important pitfall in routine immunohistochemistry-based diagnostics that pathologists should be aware of. Furthermore, the chemical rationale behind the observed misleading colour change is discussed.

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Esophageal squamous cell carcinoma with lymphoid stroma: Report of 3 cases with immunohistochemical analyses

Gastrointestinal Endoscopy ◽

10.1067/mge.2001.117154 ◽

2001 ◽

Vol 54 (4) ◽

pp. 513-517 ◽

Cited By ~ 7

Author(s):

Osamu Chino ◽

Hiroshi Kijima ◽

Hideo Shimada ◽

Kyoichi Mizutani ◽

Takayuki Nishi ◽

...

Keyword(s):

Squamous Cell Carcinoma ◽

Esophageal Squamous Cell Carcinoma ◽

Cell Carcinoma ◽

Squamous Cell ◽

Lymphoid Stroma ◽

Immunohistochemical Analyses

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Identification of renal transporters involved in sulfate excretion in marine teleost fish

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00228.2009 ◽

2009 ◽

Vol 297 (6) ◽

pp. R1647-R1659 ◽

Cited By ~ 24

Author(s):

Akira Kato ◽

Min-Hwang Chang ◽

Yukihiro Kurita ◽

Tsutomu Nakada ◽

Maho Ogoshi ◽

...

Keyword(s):

Xenopus Oocytes ◽

Proximal Tubules ◽

Border Region ◽

Marine Teleost ◽

Renal Transporters ◽

Electrode Voltage ◽

Cloned Cdna ◽

Cdna Fragments ◽

Immunohistochemical Analyses ◽

Marine Teleost Fish

Sulfate (SO42−) is the second most abundant anion in seawater (SW), and excretion of excess SO42− from ingested SW is essential for marine fish to survive. Marine teleosts excrete SO42− via the urine produced in the kidney. The SO42− transporter that secretes and concentrates SO42− in the urine has not previously been identified. Here, we have identified and characterized candidates for the long-sought transporters. Using sequences from the fugu database, we have cloned cDNA fragments of all transporters belonging to the Slc13 and Slc26 families from mefugu ( Takifugu obscurus ). We compared Slc13 and Slc26 mRNA expression in the kidney between freshwater (FW) and SW mefugu. Among 14 clones examined, the expression of a Slc26a6 paralog (mfSlc26a6A) was the most upregulated (30-fold) in the kidney of SW mefugu. Electrophysiological analyses of Xenopus oocytes expressing mfSlc26a6A, mfSlc26a6B, and mouse Slc26a6 (mSlc26a6) demonstrated that all transporters mediate electrogenic Cl−/SO42−, Cl−/oxalate2−, and Cl−/ nHCO3− exchanges and electroneutral Cl−/formate− exchange. Two-electrode voltage-clamp experiments demonstrated that the SO42−-elicited currents of mfSlc26a6A is quite large (∼35 μA at +60 mV) and 50- to 200-fold higher than those of mfSlc26a6B and mSlc26a6. Conversely, the currents elicited by oxalate and HCO3− are almost identical among mfSlc26a6A, mfSlc26a6B, and mSlc26a6. Kinetic analysis revealed that mfSlc26a6A has the highest SO42− affinity as well as capacity. Immunohistochemical analyses demonstrated that mfSlc26a6A localizes to the apical (brush-border) region of the proximal tubules. Together, these findings suggest that mfSlc26a6A is the most likely candidate for the major apical SO42− transporter that mediates SO42− secretion in the kidney of marine teleosts.

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Comparison of three intracellular markers for combined electrophysiological, morphological and immunohistochemical analyses

Journal of Neuroscience Methods ◽

10.1016/0165-0270(91)90163-t ◽

1991 ◽

Vol 38 (2-3) ◽

pp. 129-143 ◽

Cited By ~ 18

Author(s):

Jeffrey G. Tasker ◽

Neil W. Hoffman ◽

F.Edward Dudek

Keyword(s):

Immunohistochemical Analyses

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Considerations for Whole-Slide Analysis of Murine Xenografts Experiments

Journal of Histochemistry & Cytochemistry ◽

10.1369/00221554211046994 ◽

2021 ◽

Vol 69 (10) ◽

pp. 627-631

Author(s):

Abigail R. Bland ◽

John C. Ashton

Keyword(s):

Cell Death ◽

Image Analysis ◽

Cell Counting ◽

Analysis Techniques ◽

The Core ◽

Necrotic Region ◽

Areas Of Interest ◽

Image Analysis Techniques ◽

Immunohistochemical Analyses ◽

Animal Models Of Cancer

Histochemistry of tumor sections is a widely employed technique utilized to examine cell death in preclinical xenograft animal models of cancer. However, this is under the assumption that tumors are homogeneous, leading to practices such as automatic cell counting across the entire section. We have noted that in our experiments the core of the tumor is largely or partially necrotic, and lacks evidence of vascularization (in contrast to the outer areas of the tumor). We note that this can bias and confound immunohistochemical analyses that do not take care to sample areas of interest in a way to take this into account. Design-based stereology with image analysis techniques is an alternative process that could be used to measure the volume of the necrotic region compared to the volume of the whole tumor.

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