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Open AccessEditorial

Immunohistochemical Expression

by Carla Loreto and Rosario Caltabiano

Appl. Sci. 2021, 11(1), 360; https://doi.org/10.3390/app11010360 - 1 Jan 2021

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Abstract

Immunohistochemistry (IHC) is an ancillary method, widely used in pathologist practice, that allows to identify diagnostic and prognostic/predictive therapeutic response protein markers on tissue samples by the use of specific monoclonal antibodies and chromogenic substances that guarantee the visualization of the antibody–antigene binding [...] Read more.

Immunohistochemistry (IHC) is an ancillary method, widely used in pathologist practice, that allows to identify diagnostic and prognostic/predictive therapeutic response protein markers on tissue samples by the use of specific monoclonal antibodies and chromogenic substances that guarantee the visualization of the antibody–antigene binding complex under the light microscope [...] Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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9 pages, 6201 KiB

Open AccessFeature PaperArticle

Fetal Megacystis: A New Morphologic, Immunohistological and Embriogenetic Approach

by Lidia Puzzo, Giuliana Giunta, Rosario Caltabiano, Antonio Cianci and Lucia Salvatorelli

Appl. Sci. 2019, 9(23), 5155; https://doi.org/10.3390/app9235155 - 28 Nov 2019

Cited by 1 | Viewed by 3539

Abstract

Congenital anomalies of the kidney and urinary tract (CAKUT) include isolated kidney malformations and urinary tract malformations. They have also been reported in Prune-Belly syndrome (PBS) and associated genetic syndromes, mainly 13, 18 and 21 trisomy. The AA focuses on bladder and urethral [...] Read more.

Congenital anomalies of the kidney and urinary tract (CAKUT) include isolated kidney malformations and urinary tract malformations. They have also been reported in Prune-Belly syndrome (PBS) and associated genetic syndromes, mainly 13, 18 and 21 trisomy. The AA focuses on bladder and urethral malformations, evaluating the structural and histological differences between two different cases of megacystis. Both bladders were examined by routine prenatal ultrasound screening and immunohistochemistry, comparing the different expression of smooth muscular actin (SMA), S100 protein and WT1c in megacystis and bladders of normal control from fetuses of XXI gestational age. Considering the relationship between the enteric nervous system and urinary tract development, the AA evaluated S100 and WT1c expression both in bladder and bowel muscular layers. Both markers were not expressed in the bladder and bowel of PBS associated with anencephaly. In conclusion, megacystis could be considered only a macroscopic definition, concerning the size of the fetal bladder rather than the embryologic origin; it may be a single or multiple malformation; the possible association with the bowel and/or encephalic malformations will decide the outcome and prognosis in fetal megacystis. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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18 pages, 3874 KiB

Open AccessArticle

Identification of Novel Markers of Prostate Cancer Progression, Potentially Modulated by Vitamin D

by Rosario Caltabiano, Paola Castrogiovanni, Ignazio Barbagallo, Silvia Ravalli, Marta Anna Szychlinska, Vincenzo Favilla, Luigi Schiavo, Rosa Imbesi, Giuseppe Musumeci and Michelino Di Rosa

Appl. Sci. 2019, 9(22), 4923; https://doi.org/10.3390/app9224923 - 15 Nov 2019

Cited by 7 | Viewed by 2766

Abstract

Prostate cancer (PCa) is one of the most common cancers in men. The main risk factors associated with the disease include older age, family history of the disease, smoking, alcohol and race. Vitamin D is a pleiotropic hormone whose low levels are associated [...] Read more.

Prostate cancer (PCa) is one of the most common cancers in men. The main risk factors associated with the disease include older age, family history of the disease, smoking, alcohol and race. Vitamin D is a pleiotropic hormone whose low levels are associated with several diseases and a risk of cancer. Here, we undertook microarray analysis in order to identify the genes involved in PCa. We analyzed three PCa microarray datasets, overlapped all genes significantly up-regulated, and subsequently intersected the common genes identified with the down-regulated genes transcriptome of LNCaP cells treated with 1α,25(OH)₂D₃, in order to identify the common genes involved in PCa and potentially modulated by Vitamin D. The analysis yielded 43 genes potentially involved in PCa and significantly modulated by Vitamin D. Noteworthy, our analysis showed that six genes (PRSS8, SOX4, SMYD2, MCCC2, CCNG2 and CD2AP) were significantly modulated. A Pearson correlation analysis showed that five genes out of six (SOX4 was independent), were statistically correlated with the gene expression levels of KLK3, and with the tumor percentage. From the outcome of our investigation, it is possible to conclude that the genes identified by our analysis are associated with the PCa and are potentially modulated by the Vitamin D. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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16 pages, 3227 KiB

Open AccessArticle

Epigallocatechin-3-Gallate (EGCG), An Active Constituent of Green Tea: Implications in the Prevention of Liver Injury Induced by Diethylnitrosamine (DEN) in Rats

by Saleh A. Almatroodi, Mohammed A. Alsahli, Hanan Marzoq Alharbi, Amjad Ali Khan and Arshad Husain Rahmani

Appl. Sci. 2019, 9(22), 4821; https://doi.org/10.3390/app9224821 - 11 Nov 2019

Cited by 17 | Viewed by 3966

Abstract

Liver diseases are one of the most detrimental conditions that may cause inflammation, leading to tissue damage and perturbations in functions. Several drugs are conventionally available for the treatment of such diseases, but the emergence of resistance and drug-induced liver injury remains pervasive. [...] Read more.

Liver diseases are one of the most detrimental conditions that may cause inflammation, leading to tissue damage and perturbations in functions. Several drugs are conventionally available for the treatment of such diseases, but the emergence of resistance and drug-induced liver injury remains pervasive. Hence, alternative therapeutic strategies have to be looked upon. Epigallocatechin-3-gallate (EGCG) is a naturally occurring polyphenol in green tea that has been known for its disease-curing properties. In this study, we aimed to evaluate its anti-oxidative potential and protective role against diethylnitrosamine (DEN)-induced liver injury. Four different groups of rats were used for this study. The first group received normal saline and served as the control group. The second group received DEN (50 mg/kg body wt) alone and third group received DEN plus EGCG (40 mg/kg body wt) only. The fourth group were treated with EGCG only. The liver protective effect of EGCG against DEN toxicity through monitoring the alterations in aspartate transaminase (AST), and alanine transaminase (ALT) and alkaline phosphatase (ALP) activities, serum level of pro-inflammatory mediators and anti-oxidant enzymes, histopathological alterations, measurement of cellular apoptosis, and cell cycle analysis was examined. The rats that were given DEN only had a highly significantly elevated levels of liver enzymes and pro-inflammatory cytokines, highly decreased anti-oxidative enzymes, and histological changes. In addition, a significant elevation in the percentage of apoptotic nuclei and cell cycle arrest in the sub- G1 phase was detected. EGCG acts as a hepatoprotectant on DENs by reducing the serum levels of liver functional enzymes, increasing total anti-oxidative capacity, reducing pathological changes and apoptosis, as well as causing the movement of cells from the sub G1 to S or G2/M phase of the cell cycle. In conclusion, EGCG displayed a powerful hepatoprotective additive as it considerably mitigates the liver toxicity and apoptosis induced by DEN. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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11 pages, 9446 KiB

Open AccessFeature PaperArticle

Quantitative Immunomorphological Analysis of Heat Shock Proteins in Thyroid Follicular Adenoma and Carcinoma Tissues Reveals Their Potential for Differential Diagnosis and Points to a Role in Carcinogenesis

by Alessandro Pitruzzella, Letizia Paladino, Alessandra Maria Vitale, Stefania Martorana, Calogero Cipolla, Giuseppa Graceffa, Daniela Cabibi, Sabrina David, Alberto Fucarino, Fabio Bucchieri, Francesco Cappello, Everly Conway de Macario, Alberto JL Macario and Francesca Rappa

Appl. Sci. 2019, 9(20), 4324; https://doi.org/10.3390/app9204324 - 15 Oct 2019

Cited by 5 | Viewed by 2847

Abstract

Hsp27, Hsp60, Hsp70, and Hsp90 are chaperones that play a crucial role in cellular homeostasis and differentiation, but they may be implicated in carcinogenesis. Follicular neoplasms of the thyroid include follicular adenoma and follicular carcinoma. The former is a very frequent benign encapsulated [...] Read more.

Hsp27, Hsp60, Hsp70, and Hsp90 are chaperones that play a crucial role in cellular homeostasis and differentiation, but they may be implicated in carcinogenesis. Follicular neoplasms of the thyroid include follicular adenoma and follicular carcinoma. The former is a very frequent benign encapsulated nodule, whereas the other is a nodule that infiltrates the capsule, blood vessels and the adjacent parenchyma, with a tendency to metastasize. The main objective was to assess the potential of the Hsps in differential diagnosis and carcinogenesis. We quantified by immunohistochemistry Hsp27, Hsp60, Hsp70, and Hsp90 on thin sections of human thyroid tissue with follicular adenoma or follicular carcinoma, comparing the tumor with the adjacent peritumoral tissue. Hsp60, Hsp70, and Hsp90 were increased in follicular carcinoma compared to follicular adenoma, while Hsp27 showed no difference. Histochemical quantification of Hsp60, Hsp70, and Hsp90 allows diagnostic distinction between follicular adenoma and carcinoma, and between tumor and adjacent non-tumoral tissue. The quantitative variations of these chaperones in follicular carcinoma suggest their involvement in tumorigenesis, for instance in processes such as invasion of thyroid parenchyma and metastasization. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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8 pages, 1268 KiB

Open AccessArticle

Clinical and Functional Characterization of a Novel URAT1 Dysfunctional Variant in a Pediatric Patient with Renal Hypouricemia

by Blanka Stiburkova, Jana Bohata, Iveta Minarikova, Andrea Mancikova, Jiri Vavra, Vladimír Krylov and Zdenek Doležel

Appl. Sci. 2019, 9(17), 3479; https://doi.org/10.3390/app9173479 - 23 Aug 2019

Cited by 2 | Viewed by 2517

Abstract

Renal hypouricemia (RHUC) is caused by an inherited defect in the main (reabsorptive) renal urate transporters, URAT1 and GLUT9. RHUC is characterized by decreased concentrations of serum uric acid and an increase in its excretion fraction. Patients suffer from hypouricemia, hyperuricosuria, urolithiasis, and [...] Read more.

Renal hypouricemia (RHUC) is caused by an inherited defect in the main (reabsorptive) renal urate transporters, URAT1 and GLUT9. RHUC is characterized by decreased concentrations of serum uric acid and an increase in its excretion fraction. Patients suffer from hypouricemia, hyperuricosuria, urolithiasis, and even acute kidney injury. We report the clinical, biochemical, and genetic findings of a pediatric patient with hypouricemia. Sequencing analysis of the coding region of SLC22A12 and SLC2A9 and a functional study of a novel RHUC1 variant in the Xenopus expression system were performed. The proband showed persistent hypouricemia (67–70 µmol/L; ref. range 120–360 µmol/L) and hyperuricosuria (24–34%; ref. range 7.3 ± 1.3%). The sequencing analysis identified common non-synonymous allelic variants c.73G > A, c.844G > A, c.1049C > T in the SLC2A9 gene and rare variants c.973C > T, c.1300C > T in the SLC22A12 gene. Functional characterization of the novel RHUC associated c.973C > T (p. R325W) variant showed significantly decreased urate uptake, an irregular URAT1 signal on the plasma membrane, and reduced cytoplasmic staining. RHUC is an underdiagnosed disorder and unexplained hypouricemia warrants detailed metabolic and genetic investigations. A greater awareness of URAT1 and GLUT9 deficiency by primary care physicians, nephrologists, and urologists is crucial for identifying the disorder. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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10 pages, 16007 KiB

Open AccessFeature PaperArticle

Immunoexpression of Macroh2a in Uveal Melanoma

by Lucia Salvatorelli, Lidia Puzzo, Giovanni Bartoloni, Stefano Palmucci, Antonio Longo, Andrea Russo, Michele Reibaldi, Manlio Vinciguerra, Giovanni Li Volti and Rosario Caltabiano

Appl. Sci. 2019, 9(16), 3244; https://doi.org/10.3390/app9163244 - 8 Aug 2019

Cited by 7 | Viewed by 2558

Abstract

MacroH2A is a histone variant whose expression has been studied in several neoplasms, including cutaneous melanomas (CMs). In the literature, it has been demonstrated that macroH2A.1 levels gradually decrease during CM progression, and a high expression of macroH2A.1 in CM cells relates to [...] Read more.

MacroH2A is a histone variant whose expression has been studied in several neoplasms, including cutaneous melanomas (CMs). In the literature, it has been demonstrated that macroH2A.1 levels gradually decrease during CM progression, and a high expression of macroH2A.1 in CM cells relates to a better prognosis. Although both uveal and cutaneous melanomas arise from melanocytes, uveal melanoma (UM) is biologically and genetically distinct from the more common cutaneous melanoma. Metastasis to the liver is a frequent occurrence in UM, and about 40%–50% of patients die of metastatic disease, even with early diagnosis, proper treatment, and close follow-up. We wanted to investigate macroH2A.1 immunohistochemical expression in UM. Our results demonstrated that mH2A.1 expression was higher in metastatic UM (21/23, 91.4%), while only 18/32 (56.3%). UMs without metastases showed mH2A.1 staining. These data could suggest a possible prognostic role for mH2A.1 and could form a basis for developing new pharmacological strategies for UM treatment. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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12 pages, 2693 KiB

Open AccessArticle

Immunohistochemical Expression of ABCB5 as a Potential Prognostic Factor in Uveal Melanoma

by Giuseppe Broggi, Giuseppe Musumeci, Lidia Puzzo, Andrea Russo, Michele Reibaldi, Marco Ragusa, Antonio Longo and Rosario Caltabiano

Appl. Sci. 2019, 9(7), 1316; https://doi.org/10.3390/app9071316 - 29 Mar 2019

Cited by 26 | Viewed by 3396

Abstract

Uveal melanoma represents the most common primary intraocular malignancy in adults; it may arise in any part of the uveal tract, with choroid and ciliary bodies being the most frequent sites of disease. In the present paper we studied ABCB5 expression levels in [...] Read more.

Uveal melanoma represents the most common primary intraocular malignancy in adults; it may arise in any part of the uveal tract, with choroid and ciliary bodies being the most frequent sites of disease. In the present paper we studied ABCB5 expression levels in patients affected by uveal melanoma, both with and without metastasis, in order to evaluate if ABCB5 is associated with a higher risk of metastatic disease and can be used as a poor prognostic factor in uveal melanoma. The target population consisted of 23 patients affected by uveal melanoma with metastasis and 32 without metastatic disease. A high expression of ABCB5 was seen in patients with metastasis (14/23, 60.9%), compared to that observed in patients without metastasis (13/32, 40.6%). In conclusion, we found that ABCB5 expression levels were correlated with faster metastatic progression and poorer prognosis, indicating their role as a prognostic factor in uveal melanoma. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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13 pages, 5285 KiB

Open AccessFeature PaperArticle

Adapted Moderate Training Exercise Decreases the Expression of Ngal in the Rat Kidney: An Immunohistochemical Study

by Michelino Di Rosa, Paola Castrogiovanni, Francesca Maria Trovato, Lorenzo Malatino, Silvia Ravalli, Rosa Imbesi, Marta Anna Szychlinska and Giuseppe Musumeci

Appl. Sci. 2019, 9(6), 1041; https://doi.org/10.3390/app9061041 - 13 Mar 2019

Cited by 4 | Viewed by 3474

Abstract

Neutrophil gelatinase-associated lipocalin (NGAL) is a biomarker of several injuries and is upregulated in inflammatory conditions. Vitamin D was shown to have anti-inflammatory effects and to increase after physical activity. This work aimed to assess, through immunohistochemistry, the effects of an adapted moderate [...] Read more.

Neutrophil gelatinase-associated lipocalin (NGAL) is a biomarker of several injuries and is upregulated in inflammatory conditions. Vitamin D was shown to have anti-inflammatory effects and to increase after physical activity. This work aimed to assess, through immunohistochemistry, the effects of an adapted moderate training exercise (AMTE) on the expression of NGAL and vitamin D receptor (VDR) in the kidney and heart of rats. Sixteen rats were distributed into two groups: the sedentary control group and the experimental group, subjected to AMTE on the treadmill for 12 weeks. The results showed the basal expression of NGAL and VDR in both the heart and the kidney in sedentary rats; no differences in the expression of both NGAL and VDR in the heart; and a decreased NGAL and an increased VDR expression in the kidney of rats subjected to AMTE. These results suggest a possible protective role of AMTE on NGAL-associated injuries in the kidney, probably through the vitamin D signaling pathway. Our results represent an interesting preliminary data that may open new horizons in the management of NGAL-associated kidney injuries. However, further studies are needed to confirm these results and to comprehend the specific interaction between NGAL and VDR pathways in the kidney. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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27 pages, 10276 KiB

Open AccessReview

Immunohistochemical Expression of Wilms’ Tumor 1 Protein in Human Tissues: From Ontogenesis to Neoplastic Tissues

by Lucia Salvatorelli, Giovanna Calabrese, Rosalba Parenti, Giada Maria Vecchio, Lidia Puzzo, Rosario Caltabiano, Giuseppe Musumeci and Gaetano Magro

Appl. Sci. 2020, 10(1), 40; https://doi.org/10.3390/app10010040 - 19 Dec 2019

Cited by 9 | Viewed by 8201

Abstract

The human Wilms’ tumor gene (WT1) was originally isolated in a Wilms’ tumor of the kidney as a tumor suppressor gene. Numerous isoforms of WT1, by combination of alternative translational start sites, alternative RNA splicing and RNA editing, have been well documented. During [...] Read more.

The human Wilms’ tumor gene (WT1) was originally isolated in a Wilms’ tumor of the kidney as a tumor suppressor gene. Numerous isoforms of WT1, by combination of alternative translational start sites, alternative RNA splicing and RNA editing, have been well documented. During human ontogenesis, according to the antibodies used, anti-C or N-terminus WT1 protein, nuclear expression can be frequently obtained in numerous tissues, including metanephric and mesonephric glomeruli, and mesothelial and sub-mesothelial cells, while cytoplasmic staining is usually found in developing smooth and skeletal cells, myocardium, glial cells, neuroblasts, adrenal cortical cells and the endothelial cells of blood vessels. WT1 has been originally described as a tumor suppressor gene in renal Wilms’ tumor, but more recent studies emphasized its potential oncogenic role in several neoplasia with a variable immunostaining pattern that can be exclusively nuclear, cytoplasmic or both, according to the antibodies used (anti-C or N-terminus WT1 protein). With the present review we focus on the immunohistochemical expression of WT1 in some tumors, emphasizing its potential diagnostic role and usefulness in differential diagnosis. In addition, we analyze the WT1 protein expression profile in human embryonal/fetal tissues in order to suggest a possible role in the development of organs and tissues and to establish whether expression in some tumors replicates that observed during the development of tissues from which these tumors arise. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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9 pages, 204 KiB

Open AccessFeature PaperReview

Matrix Metalloproteinases and Temporomandibular Joint Disorder: A Review of the Literature

by Logan Herm, Ardit Haxhia, Flavio de Alcantara Camejo, Lobat Tayebi and Luis Eduardo Almeida

Appl. Sci. 2019, 9(21), 4508; https://doi.org/10.3390/app9214508 - 24 Oct 2019

Cited by 2 | Viewed by 2583

Abstract

Temporomandibular disorders (TMD) are progressive degenerative disorders that affect the components of the temporomandibular joint (TMJ), characterized by pain and limitations in function. Matrix metalloproteinases (MMP) are enzymes involved in physiological breakdown of tissue that can have a pathological effect from an increase [...] Read more.

Temporomandibular disorders (TMD) are progressive degenerative disorders that affect the components of the temporomandibular joint (TMJ), characterized by pain and limitations in function. Matrix metalloproteinases (MMP) are enzymes involved in physiological breakdown of tissue that can have a pathological effect from an increase in activity during inflammation. A PubMed search of the current literature (within the past 10 years) was conducted to identify human studies involving matrix metalloproteinases activity in TMJ components of patients with TMD. Two separate searches results in 34 studies, six of which met inclusion criteria. Immunohistochemistry and gene analysis were used to evaluate MMP expression in the study groups. This review showed the strongest evidence for involvement of MMP-1, MMP-2, and MMP-9 in TMD; however, limitations included low sample sizes and a lack of recent clinical studies. Future research with more definitive conclusions could allow for additional pharmaceutical targets in MMP when treating patients with temporomandibular disorders. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

15 pages, 3851 KiB

Open AccessReview

The Telocytes in the Subepicardial Niche

by Cristian Bogdan Iancu, Mugurel Constantin Rusu, Laurenţiu Mogoantă, Sorin Hostiuc and Oana Daniela Toader

Appl. Sci. 2019, 9(8), 1615; https://doi.org/10.3390/app9081615 - 18 Apr 2019

Cited by 11 | Viewed by 3421

Abstract

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 [...] Read more.

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. Full article

(This article belongs to the Special Issue Immunohistochemical Expression)

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