Acta Histochemica (ACTA HISTOCHEM)

Publications in this journal

• Article: Histological analysis of cells and matrix mineralization of new bone tissue induced in rabbit femur bones by Mg-Zr based biodegradable implants

  Sravanthi Ragamouni, Jerald Mahesh Kumar, Dolly Mushahary, Harishankar Nemani, Gopal Pande
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  ABSTRACT: The biological efficacy of bone inducing implant materials in situ can be assessed effectively by performing histological analysis. We studied the peri-implant bone regeneration around two types of biodegradable magnesium-zirconium alloys, Mg-5Zr and Mg-Zr-2Sr, using histological, histochemical and immunohistochemical methods in the femur of New Zealand White strain rabbits. Our study includes three animal groups: (a) Mg-5Zr, (b) Mg-Zr-2Sr and (c) control. In each group three animals were used and in groups ‘a’ and ‘b’ the respective alloys were implanted in cavities made at the distal ends of the femur; control animals were left without implants to observe natural bone healing. Qualitative assessment of the cellularity and matrix mineralization events of the newly formed bone tissue was done at three months after implantation by histological methods in methyl methacrylate embedded tissue without decalcifying the bone. Quantitative mineral content and density of the new bone (NB) were evaluated by the statistical analysis of dual energy X-ray absorptiometry (DXA) data obtained from three animals in each experimental group. Based on our analysis we conclude that Mg-Zr-2Sr alloy showed better osseointegration of the newly formed bone with the implant surface. Our methodology of studying peri-implant osteoinduction of degradable implants using low temperature methyl methacrylate embedding resin can be useful as a general method for determining the bio-efficacy of implant materials.
  Acta Histochemica 02/2013;
• Article: Effects of anti-phospholipid antibodies on a human trophoblast cell line (HTR-8 SV/neo)

  Jovanovic M, Bozic M, Kovacevic T, Radojcic Lj, Petronijevic M, Vicovac Lj
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  ABSTRACT: Antibodies to phospholipids (aPL) have been shown to adversely affect trophoblast invasion in vivo and in vitro. HTR-8/SVneo cells derived from first trimester of pregnancy extravillous trophoblast were studied. Matrigel invasion assay, cytochemistry and cell-based enzyme-linked immunosorbant assay (ELISA) with aPL or normal IgG was used. Our data show that aPL at 100 microg/ml decrease invasiveness of HTR-8/SVneo cells to 60% of control (p<0.01), and this was also shown for primary cytotrophoblast (to 15.5% of control, p<0.001). aPL treatment caused a significant decrease in integrin alpha(1), alpha(5), and beta(1) proteins (86%, 84%, and 87%, respectively). We conclude that HTR-8/SVneo cell culture is a suitable model to study mechanisms of action of aPL on trophoblast, which in HTR-8/SVneo cells inhibit invasion by decreasing integrins alpha(5), alpha(1), and beta(1).
  Acta Histochemica 01/2010; 112:34-41.
• Article: Topical application of aminoacids improves cutaneous wound healing in aged rat

  CORSETTI G, D’ANTONA G, DIOGUARDI SF, REZZANI R
  Acta Histochemica 01/2010; 112:497-507.
• Article: Breast adenocarcinoma MCF-7 cell line induces spontaneous osteoclastogenesis via a RANK-ligand-dependent pathway.

  Vanessa Nicolin, Roberta Bortul, Renato Bareggi, Giovanna Baldini, Bruno Martinelli, Paola Narducci
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  ABSTRACT: The metastasis of breast cancer to the skeleton is a serious clinical problem resulting in hypercalcemia, bone fragility and insurmountable pain. The invasion of bony tissue by neoplastic cells usually very rapidly affects the balance between bone apposition and bone resorption. In order to elucidate a mechanism for cancer-induced osteoclastogenesis, cells from a human breast cancer line, MCF-7, were directly co-cultured with murine monocytes RAW 264.7 type CRL 2278. Compared with controls, co-culture of MCF-7 induced differentiation of multinucleated cells by membrane-bound and soluble receptor activator of NF-kB ligand (RANKL) as quantified by ELISA, Western blot analysis, transmission electron microscopy (TEM), and immunocytochemistry. The aim of this study was to determine an in vitro model system of MCF-7 human breast cancer cells grown together with monocytes to show that expression of RANKL promotes osteoclastogenesis, which may indicate a mechanism for the development of osteolytic lesions in breast cancer bone metastasis.
  Acta Histochemica 05/2008; 110(5):388-96.
• Article: Is there a relationship between PCNA expression and diabetic placental development during pregnancy?

  Nuray Acar, Emin Turkay Korgun, Sevil Cayli, Zeliha Sahin, Ramazan Demir, Ismail Ustunel
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  ABSTRACT: We aimed to investigate the distribution pattern of proliferating cell nuclear antigen (PCNA) by immunohistochemistry and Western blot in placentas of control and diabetic rats at different stages of pregnancy. It is still not clear how proliferation is coordinated and how this coordination is affected by diabetes in the placenta. Diabetes was induced by streptozocin on the first day of pregnancy. Animals were sacrificed on days 11, 13, 17 and 21 of pregnancy. In control placentas immunolabeling intensity of PCNA was the highest on days 11 and 13 of pregnancy and decreased with progression of pregnancy. In the diabetic groups immunolabeling was less intense on days 11 and 13 of pregnancy compared to controls. However, in parallel with placental weights, PCNA immunopositivity was more intense in diabetic groups than control groups on days 17 and 21 of pregnancy, and the difference was statistically significant on day 17. According to Western blot data, on days 11 and 13 of pregnancy the amount of PCNA was greater in control groups than in the diabetics, whereas it was greater in diabetic groups than the controls on days 17 and 21 of pregnancy. We conclude that PCNA may play a role in abnormal placenta formation resulting from diabetes.
  Acta Histochemica 04/2008; 110(5):408-17.
• Article: Impact of high levels of progesterone on alpha(1)-integrin distribution in the endometrium of patients with unexplained infertility.

  Sevtap Kilic, Tahir Hatipoglu, Deniz Erdogan, Cigdem Elmas, Beril Yuksel, Nicel Tasdemir, Nafiye Yilmaz, Sertac Batioglu
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  ABSTRACT: Irregular or low expression of integrins, which are cell adhesion molecules, may be associated with infertility. We conducted a prospective controlled study evaluating the effects of supraphysiological levels of estrogen and progesterone created by human menopausal gonadotropins (HMG) and progesterone support on alpha(1)-integrin immunolocalisation in the endometrium. Three groups were enrolled in the study. The first group of patients (group 1) had unexplained infertility and had been treated with HMG and progesterone (n=27). The second group of patients (group 2) was an untreated fertile group (n=24). The third group (group 3) consisted of patients who had unexplained infertility and had received no treatment (n=11). Endometrial biopsy specimens were taken from individuals from each group during the ovulation induction period. alpha(1)-integrin immunohistochemistry was performed. Serum estradiol and progesterone levels were also measured in parallel with histological dating of endometrial biopsies. Group 1 showed no statistical difference from group 2 in alpha(1)-integrin or histological dating. Group 3 showed less alpha(1)-integrin in the glandular epithelium in the secretory phase. We observed that alpha(1)-integrin was specific to the secretory phase. Its localization was denser in group 2 when compared with group 3, which supports the conclusion that alpha(1)-integrin may be a useful marker for luteal phase quality. Moreover, the supraphysiological estrogen and progesterone levels created by HMG and progesterone support may affect the alpha(1)-integrin in the endometrium in the secretory phase in the case of unexplained infertile patients.
  Acta Histochemica 04/2008; 110(5):363-70.
• Article: Epidermal nerve fiber quantification in the assessment of diabetic neuropathy.

  Kristina K Beiswenger, Nigel A Calcutt, Andrew P Mizisin
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  ABSTRACT: Assessment of cutaneous innervation in skin biopsies is emerging as a valuable means of both diagnosing and staging diabetic neuropathy. Immunolabeling, using antibodies to neuronal proteins such as protein gene product 9.5, allows for the visualization and quantification of intraepidermal nerve fibers. Multiple studies have shown reductions in intraepidermal nerve fiber density in skin biopsies from patients with both type 1 and type 2 diabetes. More recent studies have focused on correlating these changes with other measures of diabetic neuropathy. A loss of epidermal innervation similar to that observed in diabetic patients has been observed in rodent models of both type 1 and type 2 diabetes and several therapeutics have been reported to prevent reductions in intraepidermal nerve fiber density in these models. This review discusses the current literature describing diabetes-induced changes in cutaneous innervation in both human and animal models of diabetic neuropathy.
  Acta Histochemica 04/2008; 110(5):351-62.
• Article: Late expression of FosB transcription factor in 4-aminopyridine-induced seizures in the rat cerebral cortex.

  Andrea Bagosi, Mónika Bakos, Beáta Krisztin-Péva, András Mihály
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  ABSTRACT: In this study, the immunolocalization of FosB transcription factor was investigated in acute and chronic experimental models of seizures induced by 4-aminopyridine. Wistar rats were injected intraperitoneally daily with 5mg/kg 4-aminopyridine for 1, 4, 8 and 12 days and sacrificed 24h after the last injection. Corresponding control groups received the solvent of 4-aminopyridine. Immunohistochemistry revealed an increase in FosB immunolabelling in the frontal cortex in 4-aminopyridine-treated animals compared to controls, both in acute and chronic time course groups. The dentate gyrus displayed elevated FosB immunopositivity only after repeatedly applied convulsant (4-aminopyridine), i.e. following 4, 8 and 12 days of treatment, but no significant immunolocalization was observed in the hippocampus proper. The neuronal localization of FosB after 12 days of 4-aminopyridine-induced convulsions was analysed by means of FosB-parvalbumin double immunolabelling. The increased number of double-labelled cells was significant in the frontal cortex, hilum of the dentate fascia and region CA1 of the hippocampus. We conclude that the studied neocortical and allocortical areas showed a different pattern of FosB immunolocalization, which suggests a relative deficiency of transcriptional regulation in the Ammon's horn and may be responsible for distinct response to seizure-induced cellular insult.
  Acta Histochemica 04/2008; 110(5):418-26.
• Article: Arrangement of collagen fibers in human placental stem villi.

  Leyla Sati, Ayse Yasemin Demir, Levent Sarikcioglu, Ramazan Demir
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  ABSTRACT: The aim of the study was to investigate the arrangements and related localization patterns of different collagen types in the stroma of placental stem villi by immunohistochemistry and electron microscopy. A total of 14 normal human term placental tissue samples were studied. Immunohistochemistry was performed in order to localize collagen types I, III, IV, V and cytokeratin 7 on tissue sections. Parallel tissue samples were examined by transmission electron microscopy. Semi-quantitative analysis of immunolabeling intensities was also performed to determine the distribution of fibers in stem villi stroma. All collagen types, especially collagen type V, were strongly immunopositive in the triangular areas of the stem villi stroma. However, there was no collagen type I or type III immunolabeling in the sub-trophoblastic regions. Membrane collagen type IV immunolabeling was also observed in the stroma of stem villi. Ultrastructurally, collagen fibers showed different configurations in cross, longitudinal, circular, oblique and parallel directions compared to the villous axis. We conclude that the organization of collagen fiber bundles in stem villi shows a very specific arrangement: a compact coat formed by fibrillar bundles between the vascular wall and extravascular stroma of stem villi correlated with the functional activity.
  Acta Histochemica 03/2008; 110(5):371-9.
• Article: Differential elastin and tenascin immunolabeling in the uterosacral ligaments in postmenopausal women with and without pelvic organ prolapse.

  Christian Goepel
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  ABSTRACT: Connective tissue, consisting mainly of collagen and structural glycoproteins, is an important part of the supportive structures of the genitourinary region. Relatively few data have been published with respect to the role of elastin and glycoproteins in pelvic organ prolapse (POP). Connective tissue of the uterosacral ligament in postmenopausal women with and without genital prolapse was compared. Fifty-nine consecutive women referred for hysterectomy were included in the study. The patients had POP or benign gynecological disease (e.g. myoma of the uterus). Tissue samples from the uterosacral ligament were investigated for localization and distribution of tenascin and elastin using immunofluorescence microscopy. Tissue samples of women with prolapse showed a significantly (p<0.001) weaker immunofluorescent labeling of tenascin compared to samples taken from women without prolapse. Tenascin was detectable in tissues of all women with POP, whereas its immunolabeling was decreased in the uterosacral ligament in women without POP. Intact elastin fibers were observed in tissues of all women without POP, whereas elastin was undetectable or sometimes fragmented in the uterosacral ligament in women with POP. Greater amounts of tenascin and lesser amounts of elastin were therefore found in patients with POP. These results suggest that an altered turnover of connective tissue in the uterosacral ligament might be responsible for the presence of pelvic floor relaxation in postmenopausal women. These data indicate a complex architecture of the extracellular matrix in the uterosacral ligaments, with marked differences in tenascin and elastin expression between postmenopausal women with or without POP.
  Acta Histochemica 02/2008; 110(3):204-9.
• Article: Histochemical mapping of glycoconjugates in the testis of the one humped camel (Camelus dromedarius) during rutting and non-rutting seasons.

  Ahmed Abd-Elmaksoud, Ahmed Sayed-Ahmed, Mohamed Kassab, Khaled Aly
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  ABSTRACT: In the present study, the distribution of various sugar residues in the testicular cells of sexually mature camels during rutting and non-rutting seasons was examined employing 10 fluorescein isothiocyanate- (FITC) conjugated lectins. Lectin labeling was restricted to the germ cell lines and interstitial Leydig cells, while the Sertoli cells remained completely unlabeled. Our results revealed the presence of mannose (labeled by lectins PSA, LCA), galactose (labeled by PNA), GalNAc (labeled by HPA), and GlcNAc (labeled by WGA) residues in the camel spermatogonia. However, spermatocytes were only labeled with mannose (PSA, LCA) and GlcNAc (WGA) binding lectins. Binding sites for PSA, LCA and WGA in spermatogonia and spermatocytes were only evident during the rutting season. Although spermatids were exclusively labeled with PNA in the non-rutting seasons, other lectins (PSA, GSA-I, WGA) additionally bound to camel spermatids during the rutting period. Leydig cells and basal lamina of the seminiferous tubules of camel testis were consistently labeled with the mannose- (PSA, LCA) and GlcNAc- (WGA) binding lectins in both seasons, while DBA-labeling was seen in the Leydig cells during rutting period only. In conclusion, the findings of the present study clearly indicate that the camel testis contains a wide range of glycoconjugates (bearing mannosyl, galactosyl and glucosyl residues), and they lack fucosyl residues, both in the active sexual period and in the non-breeding season. The topographical distribution of the sugar moieties in the camel testis may indicate that specific carbohydrate structures are required for spermatogenesis during periods of sexual activity.
  Acta Histochemica 02/2008; 110(2):124-33.
• Article: A comparative immunohistochemical study of endocrine cells in the digestive tract of two frugivorous bats: Artibeus cinerius and Sturnira lilium.

  Clarice Machado Dos Santos, Aparecida Alves Do Nascimento, Adriano Lucio Peracchi, Daniela Dias, Thatiana Paz Ribeiro, Armando Sales
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  ABSTRACT: The purpose of the present study was to examine the serotonin (5-hydroxytryptamine, 5-HT), gastrin (GAS), cholecystokinin (CCK) and glucagon (GLUC) endocrine cells in the gastrointestinal tract of frugivorous Phillostomidae bats, Sturnira lilium and Artibeus cinerius, to clarify the correlation between distribution of cell types and their relative frequency, with feeding habits. Five portions of the gastrointestinal tract--fundus, pilorus, and three parts of the intestine, I, II and III--were examined. Most of the immunoreactive cells in the stomach and intestine were of triangular, oval or piriform shape. Serotonin-immunoreactive cells were most commonly found in the S. lilium intestine I (66.6+/-9.9) and the A. cinerius intestine III (35+/-18). Gastrin-immunoreactive cells were the most abundant cell type in the pyloric glands of both species. They were more numerous in A. cinerius (126.9+/-27.4) than in S. lilium (75.8+/-1.8). CCK-immunoreactive cells were found in the alimentary tract epithelia at moderate frequencies in both species. GLUC-immunoreactive cells were detected at very low or low frequencies. This study suggests that there is a correlation between endocrine cell distribution and frequency, and the feeding habits of the bats.
  Acta Histochemica 02/2008; 110(2):134-42.
• Article: Hyalin is a cell adhesion molecule involved in mediating archenteron-blastocoel roof attachment.

  Edward J Carroll, Virginia Hutchins-Carroll, Catherine Coyle-Thompson, Steven B Oppenheimer
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  ABSTRACT: The US National Institutes of Health has designated the sea urchin embryo as a model organism because around 25 discoveries in this system have led to insights into the physiology of higher organisms, including humans. Hyalin is a large glycoprotein in the hyaline layer of sea urchin embryos that functions to maintain general adhesive relationships in the developing embryo. It consists of the hyalin repeat domain that has been identified in organisms as diverse as bacteria, worms, flies, mice, sea urchins and humans. Here we show, using a polyclonal antibody raised against the 11.6 S species of hyalin, that it localizes at the tip of the archenteron and on the roof of the blastocoel exactly where these two structures bond in an adhesive interaction that has been of interest for over a century. In addition, the antibody blocks the interaction between the archenteron tip and blastocoel roof. These results, in addition to other recent findings from this laboratory that will be discussed, suggest that hyalin is involved in mediating this cellular interaction. This is the first demonstration that suggests that hyalin functions as a cell adhesion molecule in many organisms, including humans.
  Acta Histochemica 02/2008; 110(4):265-75.
• Article: Immunolocalization of farnesoid X receptor (FXR) in mouse tissues using tissue microarray.

  Hiroyuki Higashiyama, Mine Kinoshita, Satoshi Asano
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  ABSTRACT: Farnesoid X receptor (FXR) is a member of the nuclear receptor family and is known to play important roles in bile acid homeostasis, and lipid and glucose metabolism. In this study, to elucidate the systemic physiological functions of FXR, comprehensive immunohistochemical analysis of cell/subcellular localization of FXR and its heterodimer partner, retinoid X receptor (RXR)-alpha, in adult mice tissues was performed using tissue microarray (TMA)-based immunohistochemistry. FXR immunolabeling was observed in the enterohepatic system--including absorptive epithelium in the intestines, hepatocytes and gall bladder epithelium, several epithelial lineage cells including the basal cells of stratified epithelium in the tongue, esophagus, forestomach--skin, corneal epithelium and ciliary body epithelium in the eye and adrenocortical cells--including glandular cells in the zona reticularis/fasciculata. In these FXP-positive cells, FXR was preferentially localized to the nucleus. RXR-alpha was ubiquitously distributed in the nucleus of most cell types, including FXR-positive cell types in the examined tissues. These data suggest that FXR might have various physiological roles, not only in bile acid homeostasis, and lipid and glucose metabolism, but also in the epithelial cell barrier, visual and urinary function through multiple organ systems.
  Acta Histochemica 02/2008; 110(1):86-93.
• Article: Quantitative analysis of ciliary ultrastructure in patients with primary ciliary dyskinesia.

  Serap Sirvanci, Z Seda Uyan, Feriha Ercan, Bulent Karadag, Refika Ersu, Fazilet Karakoc, Elif Dagli, Tangul San
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  ABSTRACT: The present study was designed to investigate dynein arm and microtubule defects quantitatively in patients with respiratory disease and to establish the clinical relevance of dynein arm deficiency and microtubule abnormalities. Thirty-four patients with recurrent upper and/or lower respiratory infections were included in the study. Nasal mucosal brushings were fixed in glutaraldehyde and routine electron microscopic procedures were carried out. At least 20 cross-sectioned cilia were examined from each subject. Dynein arm and microtubular abnormalities were quantified and a statistical analysis was performed. Twenty-nine percent of the patients showed dynein arm deficiency and a further 21% had possible deficiency (PD). Microtubule defects in patients with dynein arm deficiency and PD were found to be significantly increased compared to the patients with no dynein arm deficiency. The most prominent defect in the dynein arm deficiency group was a translocation of central and/or peripheral microtubules. The high percentage of translocation defect in this group of patients suggests that these defects are primary, rather than secondary to infection.
  Acta Histochemica 02/2008; 110(1):34-41.
• Article: Expression and regulation of Sef, a novel signaling inhibitor of receptor tyrosine kinases-mediated signaling in the nervous system.

  Claudia Grothe, Peter Claus, Kirsten Haastert, Ela Lutwak, Dina Ron
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  ABSTRACT: Fibroblast growth factors (FGFs) signal via four distinct high affinity cell surface tyrosine kinase receptors, termed FGFR1-FGFR4 (FGFR-FGF-receptor). Recently, a new modulator of the FGF signaling pathway, the transmembrane protein 'similar expression to FGF genes' (Sef), has been identified in zebrafish and subsequently in mammals. Sef from mouse and human inhibits FGF mitogenic activity. In the present study, we analyzed the expression of Sef in distinct rat brain areas, in the spinal cord and in peripheral nerves and spinal ganglia using semi-quantitative RT-PCR. Furthermore, we studied the cellular expression pattern of Sef in intact spinal ganglia and sciatic nerves and, in addition, after crush lesion, using in situ hybridization and immunohistochemistry. Sef transcripts were expressed in all brain areas evaluated and in the spinal cord. A neuronal expression was found in both intact and injured spinal ganglia. Intact sciatic nerves, however, showed little or no Sef expression. Seven days after injury, high Sef expression was concentrated to the crush site, and Schwann cells seemed to be the source of Sef. The labeling pattern of up-regulated Sef was complementary to the patterns of FGF-2 and FGFR1-3, which were localized proximal and distal to the crush site. These results suggest an involvement of Sef during the nerve regeneration process, possibly by fine-tuning the effects of FGF signaling.
  Acta Histochemica 02/2008; 110(2):155-62.