Histology and histopathology Vol.32,nº11 (2017)

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Open Access
Effect of estradiol on the expression of angiogenic factors in epithelial ovarian cancer
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Valladares, Macarena; Plaza Parrochia, Francisca; Lépez, Macarena; López, Daniela; Gabler, Fernando; Gayan, Patricio; Selman, Alberto; Vega, Margarita; Romero, Carmen
Introduction: Ovarian cancer presents a high angiogenesis (formation of new blood vessels) regulated by pro-angiogenic factors, mainly vascular endothelial growth factor (VEGF) and nerve growth factor (NGF). An association between endogenous levels of estrogen and increased risk of developing ovarian cancer has been reported. Estrogen action is mediated by the binding to its specific receptors (ERα and ERβ), altered ERα/ERβ ratio may constitute a marker of ovarian carcinogenesis progression. Objective: To determine the effect of estradiol through ERα on the expression of NGF and VEGF in epithelial ovarian cancer (EOC). Methodology: Levels of phosphorylated estrogen receptor alpha (pERα) were evaluated in well, moderate and poorly differentiated EOC samples (EOC-I, EOC-II, EOC-III). Additionally, ovarian cancer explants were stimulated with NGF (0, 10 and 100 ng/ml) and ERα, ERβ and pERα levels were detected. Finally, human ovarian surface epithelial (HOSE) and epithelial ovarian cancer (A2780) cell lines were stimulated with estradiol, where NGF and VEGF protein levels were evaluated. Results: In tissues, ERs were detected being pERα levels significantly increased in EOC-III samples compared with EOC-I (p<0.05). Additionally, ovarian explants treated with NGF increased pERα levels meanwhile total ERα and ERβ levels did not change. Cell lines stimulated with estradiol revealed an increase of NGF and VEGF protein levels (p<0.05). Conclusions: Estradiol has a positive effect on pro-angiogenic factors such as NGF and VEGF expression in EOC, probably through the activation of ERα; generating a positive loop induced by NGF increasing pERα levels in epithelial ovarian cells.
Open Access
Histopathological features of endometritis eosinophilica in mares
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Grimm, Anna Lena; Schoon, Heinz Adolf; Schöniger, Sandra
Equine endometritis eosinophilica (EE) is rarely described and its diagnostic criteria are not defined. The aim of this study was to characterize histological features of EE. A data base (1995-2013) was searched for biopsies with increased eosinophils. This study included all biopsies with this diagnosis and representative biopsies without this record. The definition of equine EE was based on criteria for EE in women and the results of the determination of physiological numbers of eosinophils within the equine endometrium. EE was diagnosed in 55 mares. Biopsies of 10 mares contained eosinophils exceeding the physiological range, but no EE; the diagnosis of eosinophilic infiltrates (EI) was applied. Those of the remaining mares (n=126) displayed eosinophils within the physiological range (EWPR). An irregular glandular differentiation during the breeding season was detected in 25% of mares with EE, 33% of mares with EI and 24% of the mares with EWPR. Most mares with EE (93%), EI (90%) and EWPR (72%) showed endometrosis; it was high grade in 11% with EE and 7% with EWPR. Endometritis was diagnosed within 56% of mares with EE, 40% of mares with EI and 37% of those with EWPR. In mares with EE suppurative endometritis dominated (58%) and in those with EWPR nonsuppurative endometritis (58%). This study indicates EE as a primary fertility reducing disease. Results suggest an association between eosinophilic infiltration and the presence of neutrophils. Further, they provide the basis for future studies into the pathogenesis, prognosis and therapy of EE.
Open Access
Assessment of immunologic, proangiogenic and neurogenic properties of human peripheral nerve epineurium for potential clinical application
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Klimczak, Aleksandra; Siemionow, Maria; Futoma, Katarzyna; Jundzill, Arkadiusz; Patrzalek, Dariusz
The epineural sheath is a promising naturally occurring material for enhancement of peripheral nerve regeneration. Based on a literature search there is a limited number of reports on the biological and immunological properties of human epineurium. The goal of this study was to assess, using immunocytochemical methods, the immunological (HLA class I and II antigens, T lymphocytes, macrophages), proangiogenic (VEGF, CD31), and neurogenic (GFAP, S-100) properties of human epineurium isolated from ilioinguinal nerves (n=19) taken from deceased donors, and from sciatic nerves (n=12) taken from limbs amputated due to critical ischemia. Our studies confirmed reduced expression of HLA class II antigens on the infiltrating cells, a reduced number of T lymphocytes, and greater vessel density in the epineurium obtained from deceased organ donors. Macrophages were more abundant in the epineurium isolated from the amputated limbs. We found that the epineurium harvested from peripheral nerves of the deceased donors showed negligible immunogenic and increased proangiogenic properties compared to the epineurium of nerves taken from amputated limbs. These findings support the rationale to use human epineurium obtained from deceased donors as a new biological material for enhancement of peripheral nerve repair for potential clinical application in regenerative medicine.
Open Access
Neurons other than motor neurons in motor neuron disease
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Ruffoli, Riccardo; Biagioni, Francesca; Busceti, Carla L.; Gaglione, Anderson; Ryskalin, Larisa; Gambardella, Stefano; Frati, Alessandro; Fornai, Francesco
Amyotrophic lateral sclerosis (ALS) is typically defined by a loss of motor neurons in the central nervous system. Accordingly, morphological analysis for decades considered motor neurons (in the cortex, brainstem and spinal cord) as the neuronal population selectively involved in ALS. Similarly, this was considered the pathological marker to score disease severity ex vivo both in patients and experimental models. However, the concept of non-autonomous motor neuron death was used recently to indicate the need for additional cell types to produce motor neuron death in ALS. This means that motor neuron loss occurs only when they are connected with other cell types. This concept originally emphasized the need for resident glia as well as non-resident inflammatory cells. Nowadays, the additional role of neurons other than motor neurons emerged in the scenario to induce non-autonomous motor neuron death. In fact, in ALS neurons diverse from motor neurons are involved. These cells play multiple roles in ALS: (i) they participate in the chain of events to produce motor neuron loss; (ii) they may even degenerate more than and before motor neurons. In the present manuscript evidence about multineuronal involvement in ALS patients and experimental models is discussed. Specific sub-classes of neurons in the whole spinal cord are reported either to degenerate or to trigger neuronal degeneration, thus portraying ALS as a whole spinal cord disorder rather than a disease affecting motor neurons solely. This is associated with a novel concept in motor neuron disease which recruits abnormal mechanisms of cell to cell communication.
Open Access
Sustanon induces dose-independent hypertrophy and satellite cell proliferation in slow oxidative fibers of avian skeletal muscle
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Allouh, Mohammed Z.; Jarrar, Ahmad A.; Asfour, Hasan A.; Said, Raed S.; Shaqoura, Emad I.
Sustanon is a well-known anabolic drug that is used to treat hypogonadism and restore muscle mass and bone density. As research to date has been limited to its effects in glycolytic fibers, this study aimed to investigate the dose-related effects of Sustanon on the oxidative fibers of avian skeletal muscle. Adult female chickens were randomly divided into 4 groups: control (C), received a dose of 100 μl normal saline per injection; and Sustanon-1, -2, and -3 (S1, S2, and S3), that received a dose of 12.5, 25, or 50 mg/kg Sustanon per injection, respectively. Each bird received 4 injections at weekly intervals (1 injection/week). Robust histochemical and immunofluorescent techniques along with morphometric analyses were applied to determine the oxidative activity and morphological variations of the oxidative muscle fibers in all groups. Sustanontreated groups exhibited significant increases in fiber size and numbers of satellite cells and myonuclei compared to the control group. However, no significant variations were found between Sustanon-treated groups in the aforementioned indices. In conclusion, Sustanon induced oxidative fiber hypertrophy that was associated with satellite cell proliferation and myonuclear accretion in avian skeletal muscle. Furthermore, the effects of Sustanon appeared to be dose-independent.
Open Access
Are non-muscle actin isoforms functionally equivalent?
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Simiczyjew, Aleksandra; Pietraszek Gremplewicz, Katarzyna; Mazur, Antonina Joanna; Nowak, Dorota
Actin is highly conserved and it is the most widespread protein in eukaryotic cells. One of the most important features of actin, which allows it to have many different functions, is its ability to polymerize and interact with many other proteins. Actins are the major constituent of the actin cytoskeleton, which is an important system that is involved in various aspects of cell function, including cell motility, structure, integrity, regulation of signal transduction and transcription. Six mammal actin isoforms are highly conserved and share common functions. Two of them, β and γ non-muscle actin isoforms, which differ only by four amino acids located at the N-terminus of the polypeptide chain, are required for survival and proper cell functioning. We also summarized data about actbl2, which is suggested to be a newly discovered isoactin. Here, we review the current knowledge about tissue-specific expression of the non-muscle actin isoforms and possible functional differences between them. We also discuss molecular tools, which in recent years have allowed for a better understanding of the role of these proteins in cell functioning.
Open Access
Evolutionary trade-offs in kidney injury and repair
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Lei, Yutian; Anders, Hans Joachim
Evolutionary medicine has proven helpful to understand the origin of human disease, e.g. in identifying causal roles of recent environmental changes impacting on human physiology (environmentphenotype mismatch). In contrast, diseases affecting only a limited number of members of a species often originate from evolutionary trade-offs for usually physiologic adaptations assuring reproductive success in the context of extrinsic threats. For example, the G1 and G2 variants of the APOL1 gene supporting control of Trypanosoma infection come with the trade-off that they promote the progression of kidney disease. In this review we extend the concept of evolutionary nephrology by discussing how the physiologic adaptations (danger responses) to tissue injury create evolutionary trade-offs that drive histopathological changes underlying acute and chronic kidney diseases. The evolution of multicellular organisms positively selected a number of danger response programs for their overwhelming benefits in assuring survival such as clotting, inflammation, epithelial healing and mesenchymal healing, i.e. fibrosis and sclerosis. Upon kidney injury these danger programs often present as pathomechanisms driving persistent nephron loss and renal failure. We explore how classic kidney disease entities involve insufficient or overshooting activation of these danger response programs for which the underlying genetic basis remains largely to be defined. Dissecting the causative and hierarchical relationships between danger programs should help to identify molecular targets to control kidney injury and to improve disease outcomes.
Open Access
Sialic acid expression in human fetal skeletal muscle during limb early myogenesis
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Marini, Mirca; Sarchielli, Erica; Zappoli Thyrion, Giorgia Donata; Ambrosini, Stefano; Sgambati, Eleonora
Investigations on animal models demonstrated that changes of sialic acid (SA) expression, particularly the polymeric form, in the skeletal muscle during embryonic and post-natal development seem to be related to muscle differentiation and functionality onset. The aim of this study was to evaluate the monomeric and polymeric SA expression in human skeletal muscle during early stages of fetal development, when important morphofunctional events occur. Specimens of fetal skeletal muscle from limb, between 9 and 12 weeks of gestation (wg), were obtained from 19 pregnant women. To investigate some morphofunctional features occurring during this development period, haematoxylin-eosin staining, tunel assay and immunohistochemistry for connexin-43 (Cx43) and parvalbumin were performed. SA expression and characterization was evaluated using lectin histochemistry (MAA, SNA, PNA, SBA, DBA), associated with enzymatic and chemical treatments. Polysialic acid (PSA) expression was also evaluated using immunohistochemistry. The results showed apoptotic myotubes between 9 and 10.5 wg, disappearing from 11 wg; Cx43 was more abundant in myotubes/myoblasts between 9 and 9.5 wg, decreasing and/or disappearing from 10 wg and parvalbumin was present in myotubes between 10 and 10.5 wg. PSA was revealed in myotubes/myoblasts from 9 to 10.5 wg; from 11 wg it was reduced or disappeared. Monomeric SA appeared in myotubes/myoblasts from 10 wg, increasing successively; acetylated SA was present from 11 wg. These findings demonstrated that changes in expression of various types of SA, occurring in human fetal skeletal muscle during early development, seem to be related to some morphofunctional aspects distinctive of this organogenesis crucial period.
Open Access
Adrenal cyst with both Müllerian and mesothelial differentiation - a clinicopathological and immunohistochemical study with implications for histogenesis
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Kopersk, Łukasz i; Szczepankiewicz, Benedykt; Pihowicz, Paweł; Fus, Łukasz; Wolińska, Ewa; Górnicka, Barbara
True epithelial-lined cysts are rare forms of adrenal cystic lesions, the pathogenesis of which is still not fully understood. In this report we present a case of an adrenal cyst diagnosed incidentally on imaging in a 31-year-old, previously healthy, obese woman. Due to non-specific hormonal disorders and enlargement of the lesion, a right-sided laparoscopic adrenalectomy was performed. The cyst was lined predominantly by ciliated cuboidal-to-columnar, Müllerian-type epithelium, and focally by flat-to-cuboidal, mesothelium-like lining. Immunohistochemistry demonstrated a strong positive reaction in the cells of both types of lining for CKAE1/AE3, CK19, CK7 and WT1, and both had a negative reaction for CK20, CD34, Melan-A, SF1, TTF1, SMA and CDX2. The cells of the ciliated cuboidal-to-columnar epithelium were strongly positive for PAX8, ER, Ep-CAM and EMA, focally positive for PR, and were negative for calretinin, whereas the cells of the flat-to-cuboidal lining were positive for calretinin and podoplanin and showed only a weak positive response in individual cells for PAX8, EMA and EpCAM, but were negative for ER and PR. This is the first reported case of an adrenal ciliated epithelial cyst with Müllerian differentiation (confirmed immunohistochemically) in the English literature. The differences in morphology and immunophenotype of the two types of lining (epithelial Müllerian phenotype versus mesothelial phenotype), suggest that some adrenal epithelial cysts probably form due to metaplasia of mesothelium-derived lining. A similar mechanism may also be involved in the pathogenesis of at least some of the so-called Müllerian cysts (or inclusions) in other locations
Open Access
Expression of DNA methylation-related proteins in invasive lobular carcinoma of breast: comparison to invasive ductal carcinoma
(Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Cha, Yoon Jin; Kim, Hye Min; Koo, Ja Seung
Purpose: We aimed to compare the expression of DNA methylation-related proteins in invasive lobular carcinoma (ILC) of breast with those of invasive ductal carcinoma (IDC) of breast and to assess its potential clinical application. Methods: Immunohistochemical staining of DNA methylation-related proteins (5-meC, DNMT1, DNMT3B and ISL-1) was applied to tissue microarrays generated from 108 ILCs and 203 IDCs. Protein expression and its correlation with clinicopatholgic variables were statistically analyzed. Results: ISL-1 and DNMT3B were highly expressed in ILC (p<0.001) and tumoral 5-meC was highly expressed in IDC (p=0.006). DNMT1 (p<0.001) showed higher expression rate in luminal A type ILC. ISL-1 and DNMT3B showed higher expression rate in both luminal A type and luminal B type of ILC (p<0.05). In IDC, tumoral 5-meC commonly showed high positivity (p=0.039). On univariate analysis, shorter disease-free survival of ILC was associated with DNMT1 high positivity (p=0.001) and ISL-1 positivity (p=0.018). Conclusion: DNA methylation-related proteins are differentially expressed in ILC and IDC, and DNMT1, DNMT3B and ISL-1 show high expression rate in ILC.

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