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  • Expression and localization of meiosis-associated protein in gonads of female rats at different stages
    Acta Histochem. (IF 1.719) Pub Date : 2020-01-18
    Yihui Liu; Xiaorui Fan; Meishan Yue; Weidong Yue; Xinrong Zhang; Jingwen Zhang; Gaoya Ren; Junping He

    It was well known that a critical process of oogenesis in the female mammalian was the entry of mitotic oogonia into meiosis. Early studies from model animal mice suggested that the retinoic acid (RA) response signal protein STRA8 (stimulated by retinoic acid gene 8) and the meiosis-specific chromosomal behavior marker protein SCP3 (Synaptonemal Complex Protein 3) were two crucial molecular markers during meiosis. The expression of STRA8 and SCP3 at different stages in rat ovaries was investigated by immunohistochemistry, qRT-PCR and Western Blot. Immunohistochemistry results showed that STRA8 and SCP3 were mainly expressed in embryonic stage. And STRA8 was expressed in the cytoplasm and nucleus of the ovaries after birth. qRT-PCR and Western Blot results showed that the mRNA and protein levels of STRA8 and SCP3 were expressed in embryonic stage. The expression of STRA8 and SCP3 indicated germ cells enter meiosis in rats embryo, and STRA8 and SCP3 could serve as molecular markers for the meiosis in rats. The localization of STRA8 in the nucleus increased the possibility that STRA8 might act as transcription factor or activate transcription to function after birth.

    更新日期:2020-01-21
  • Utility of the immunohistochemical analysis of DNA mismatch-repair proteins in endometrial hyperplasia
    Acta Histochem. (IF 1.719) Pub Date : 2020-01-17
    Nabiha Missaoui; Nesrine Boukhari; Sarra Limam; Sihem Hmissa; Moncef Mokni

    The utility of the expression lack of DNA mismatch-repair (MMR) proteins in the detection of Lynch syndrome in endometrial hyperplasia as precursor lesion of endometrial carcinoma has not been well-established. The study investigated the immunoexpression pattern of MMR proteins in endometrial hyperplasia from Tunisian patients. We carried out a retrospective study of 60 endometrial hyperplasias diagnosed among Tunisian patients. Expression of MLH1, MSH2, MSH6, and PMS2 proteins was performed by immunohistochemistry on whole-slide sections of archival tissues. Analysis of MLH1 promoter methylation and microsatellite alterations was conducted in appropriate cases. Microsatellite instability screening was assessed using the Bethesda panel, including BAT25, BAT26, D17S250, D2S123, and D5S346 markers. Expression of MMR proteins was observed in all hyperplasias without atypia as well as in 27 out of 29 atypical hyperplasias. Only two atypical hyperplasias exhibited expression loss of MMR proteins. A single case revealed MSH6 expression lack. Expression loss of MLH1 and PMS2 was identified in another atypical hyperplasia and was associated with hypermethylation of MLH1 promoter. This patient had no familial history of endometrial cancer at the diagnostic time. The two deficient MMR cases showed microsatellite stable pattern. In conclusion, only two endometrial hyperplasias displayed an altered pattern of MMR expression. Our results suggest the limited utility of the immunohistochemical analysis of MMR protein in the early detection of Lynch syndrome in Tunisian patients diagnosed with endometrial hyperplasias. Multicenter studies with larger sample size are needed to more explore these findings.

    更新日期:2020-01-17
  • Eicosapentaenoic and docosapentaenoic acids lessen the expression of PPARγ/Cidec affecting adipogenesis in cultured 3T3-L1 adipocytes
    Acta Histochem. (IF 1.719) Pub Date : 2020-01-17
    Fabiane Ferreira Martins; Marcia Barbosa Aguila; Carlos Alberto Mandarim-de-Lacerda

    Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) have benefits in the metabolism of adipose tissue. However, its contribution to the adipogenesis is not entirely elucidated. The study aimed to evaluate the effects of EPA and DHA on adipogenesis, especially in the PPARγ (peroxisome proliferator-activated receptor-gamma) and Cidec (cell death-inducing DFFA-like effector c) pathway. Twenty-four hours after confluence, 3T3-L1 adipocytes were treated with EPA (100 μM), DHA (50μM) and EPA (100μM) + DHA (50μM) and at the end of differentiation (day 11) the cells were collected for analysis. Cell viability analysis indicated that the concentrations used for EPA and DHA did not cause cytotoxicity in cultured 3T3l1 adipocytes. The treatments have lessened the triacylglycerol accumulation in the adipocyte cytoplasm that, compared to the control group, were EPA-32%, DHA-38%, EPA + DHA −24%. The double-labeling immunofluorescence showed a signal attenuation of protein expressions of PPARγ, CIDEC, and SREBP-1c (sterol regulatory element-binding protein). EPA and DHA had a significant impact on the expression of cleaved CASPASE 3, which increases cell apoptosis and gene expressions of Pparγ and Cidec in the treated groups. Also, there was a reduction of C/ebpα (CCAAT/enhancer-binding protein alpha), Cd36 (cluster differentiation 36), and Foxo1 (forkhead box O). In conclusion, the study determined the ability of both EPA and DHA, alone or combined, in the adipogenesis modulation in cultured 3T3-L1 adipocytes, affecting the cell differentiation, maturation, and consequently, reducing adipogenesis via PPARγ-CIDEC suppression.

    更新日期:2020-01-17
  • Differences in the expression of catecholamine-synthesizing enzymes between vesicular monoamine transporter 1- and 2-immunoreactive glomus cells in the rat carotid body
    Acta Histochem. (IF 1.719) Pub Date : 2020-01-16
    Kouki Kato; Takuya Yokoyama; Tatsumi Kusakabe; Katsuhiko Hata; Seigo Fushuku; Nobuaki Nakamuta; Yoshio Yamamoto

    Vesicular monoamine transporters (VMAT) 1 and 2 are responsible for monoamine transportation into secretary vesicles and are tissue-specifically expressed in central and peripheral monoaminergic tissues, including the carotid body (CB). The aim of the present study was to examine the expression of catecholamine-synthesizing enzymes in VMAT1- and VMAT2-immunoreactive glomus cells in the rat CB using multiple immunolabeling. The expression of VMAT1 and VMAT2 mRNA in the CB was confirmed by RT-PCR. Immunohistochemistry revealed that VMAT1 immunoreactivity was predominant in glomus cells rather than VMAT2 immunoreactivity. Glomus cells with VMAT1 immunoreactivity exhibited weak/negative VMAT2 immunoreactivity, and vice versa. Immunoreactivities for VMAT1 and tyrosine hydroxylase, the rate-limiting enzyme for catecholamine biosynthesis, were co-localized in the same glomus cells and a positive correlation was confirmed between the two immunoreactivities (Spearman’s coefficient = 0.82; p < 0.05). Although some glomus cells showed co-localization of VMAT2 and dopamine β-hydroxylase immunoreactivity, the biosynthetic enzyme for noradrenaline, VMAT2 immunoreactivity appeared to be less associated with both catecholamine-synthesizing enzymes as indicated by a correlation analysis (TH: Spearman’s coefficient = 0.38, DBH: Spearman’s coefficient = 0.26). These results indicate that heterogeneity on functional role would exist among glomus cells in terms of VMAT isoform and catecholamine-synthesizing enzymes expression.

    更新日期:2020-01-17
  • 更新日期:2020-01-16
  • Ultrastructural characterization of vitamin D receptors and metabolizing enzymes in the lipid droplets of the fatty liver in rat
    Acta Histochem. (IF 1.719) Pub Date : 2020-01-10
    Filipović Natalija; Bočina Ivana; Restović Ivana; Grobe Maximilian; Kretzschmar Genia; Kević Nives; Mašek Tomislav; Vitlov Uljević Marija; Jurić Marija; Vukojević Katarina; Saraga-Babić Mirna; Vuica Ana

    Vitamin D is a steroid hormone with numerous actions in the organism. There are strong evidences that relate vitamin D deficiency with liver lipid metabolism disturbances, but the mechanism of this action is still unknown. In our previous work we postulated the localization and accumulation of vitamin D receptor (VDR) in membrane of the lipid droplets (LDs) in hepatocytes. In this study, we applied the transmission electron microscopy (TEM) to confirm this hypothesis by using a long-term (6 months) high sucrose intake rat model that was previously found to be appropriate for research of the hepatic lipid accumulation. In addition to the VDR, we also found key vitamin D metabolizing enzymes, 1α-hydroxylase and CYP 24 associated with the membrane of the LDs. A light-microscopy data revealed significant increase in expression of VDR and CYP 24 in liver of high-sucrose treated rats, in comparison to controlones. According to the best of our knowledge, this is a first study confirming the presence of the VDR in the membrane of the LDs in general and also in particular in LDs of the hepatocytes that were accumulated as a consequence of the prolonged high sucrose intake. Moreover, we found association of main vitamin D metabolizing enzymes with LD membrane. These results provide a new insight in the possible relation of vitamin D signalling system with LD morphology and function and with the lipid metabolism in general.

    更新日期:2020-01-10
  • Histochemical and immunohistochemical localization of nitrergic structures in the carotid body of spontaneously hypertensive rats
    Acta Histochem. (IF 1.719) Pub Date : 2020-01-07
    Dimitrinka Y. Atanasova; Angel D. Dandov; Nikolay D. Dimitrov; Nikolai E. Lazarov

    The carotid body (CB) is a multipurpose metabolic sensor that acts to initiate cardiorespiratory reflex adjustments to maintain homeostasis of blood-borne chemicals. Emerging evidence suggests that nitric oxide increases the CB chemosensory activity and this enhanced peripheral chemoreflex sensitivity contributes to sympathoexcitation and consequent pathology. The aim of this study was to examine by means of NADPH-diaphorase histochemistry and nitric oxide synthase (NOS) immunohistochemistry the presence and distribution of nitrergic structures in the CB of spontaneously hypertensive rats (SHRs) and to compare their expression patterns to that of age-matched normotensive Wistar rats (NWRs). Histochemistry revealed that the chemosensory glomus cells were NADPH-d-negative but were encircled by fine positive varicosities, which were also dispersed in the stroma around the glomeruli. The NADPH-d-reactive fibers showed the same distributional pattern in the CB of SHRs, however their staining activity was weaker when compared with NWRs. Thin periglomerular, intraglomerular and perivascular varicose fibers, but not glomus or sustentacular cells in the hypertensive CB, constitutively expressed two isoforms of NOS, nNOS and eNOS. In addition, clusters of glomus cells and blood vessels in the CB of SHRs exhibited moderate immunoreactivity for the third known NOS isoenzyme, iNOS. The present study demonstrates that in the hypertensive CB nNOS and eNOS protein expression shows statistically significant down-regulation whereas iNOS expression is up-regulated in the glomic tissue compared to normotensive controls. Our results suggest that impaired NO synthesis could contribute to elevated blood pressure in rats via an increase in chemoexcitation and sympathetic nerve activity in the CB.

    更新日期:2020-01-07
  • Use of synthetic polymers improves the quality of vitrified caprine preantral follicles in the ovarian tissue
    Acta Histochem. (IF 1.719) Pub Date : 2020-01-02
    Diego Alberto Montano Vizcarra; Yago Pinto Silva; Jamily Bezerra Bruno; Danielle Cristina Calado Brito; Deysi Dipaz Berrocal; Luciana Mascena Silva; Maria Luana Gaudencia dos Santos Morais; Benner Gerardo Alves; Kele Amaral Alves; Francielli Weber Santos Cibin; José Ricardo Figueiredo; Mary B. Zelinski; Ana Paula Ribeiro Rodrigues

    The aim of this study was to evaluate whether the addition of synthetic polymers to the vitrification solution affected follicular morphology and development and the expression of Ki-67, Aquaporin 3 (AQP3) and cleaved Caspase-3 proteins in ovarian tissue of the caprine species. Caprine ovaries were fragmented and two fragments were immediately fixed (Fresh Control) for morphological evaluation, while other two were in vitro cultured for 7 days (Cultured Control) and fixed as well. The remaining fragments were distributed in two different vitrification groups: Vitrified and Vitrified/Cultured. Each group was composed of 4 different treatments: 1) Sucrose (SUC); 2) SuperCool X-1000 0.2 % (X-1000); 3) SuperCool Z-1000 0.4 % (Z-1000) or 4) with polyvinylpyrrolidone K-12 0.2 % (PVP). Also, Fresh Control, Cultured Control, SUC and X-1000 were destined to immunohistochemical detection of Ki-67, AQP3 and cleaved Caspase-3 proteins. Morphologically, the treatment with X-1000 showed no significant difference with the Fresh Control group and was superior to the other treatments. After the cleaved caspase-3 analysis, X-1000 showed the lowest percentages of strong immunostaining while Cultured Control showed the highest. Also, a positive correlation was found between the percentages of degenerated follicles and the percentages of strong staining intensity follicles. Regarding the AQP3 analysis, the highest percentages of strong AQP3 staining intensity were found in X-1000. In conclusion, we have demonstrated that the addition of the synthetic polymer SuperCool X-1000 to the vitrification solution improved the current vitrification protocol of caprine ovarian tissue.

    更新日期:2020-01-02
  • Expression and localization of endogenous phospholipase Cβ3 confined to basal cells in situ of immature ducts and adult excretory ducts of submandibular gland of mice
    Acta Histochem. (IF 1.719) Pub Date : 2019-12-30
    Atsara Rawangwong; Masahiko Watanabe; Hisatake Kondo; Wiphawi Hipkaeo

    Our previous study demonstrated that, different from the parotid and sublingual glands, the submandibular glands of adult mice did not show an immunoblot band for PLCβ3 which is critical in the secretion mechanism by muscarinic cholinergic signaling. Therefore, the submandibular glands of mice at various stages of postnatal development were examined for this enzyme molecule in immunoblot and immunohistochemistry. In immunoblot, a weak band for PLCβ3-expression was detected only at early postnatal stages. In immunohistochemistry, PLCβ3-immunoreactivity was distinctly found in most basally located cells of immature ducts, while the immunoreactivity was weakly seen in terminal tubule cells without significant immunoreactivity in adjacent acinar cells. In contrast, the immunoreactivity was distinctly found in some basal cells of adult excretory ducts, and it was ultrastructurally localized densely in close association with bundles of tonofilaments in the cells. The present finding suggests the possibility that Ca2+ signaling governed by phospholipase Cβ3 is involved in the differentiation of ductal basal cells into apical cells through control of keratin molecule(s) in the cells.

    更新日期:2019-12-30
  • Age-related changes in the articular cartilage of the mandible of rats
    Acta Histochem. (IF 1.719) Pub Date : 2019-12-28
    M.F. Calderon; R.A.B. Nucci; R.R. de Souza

    This study investigated the effects of aging on the articular cartilage of the mandible. Wistar rats were divided in two groups (n = 10/per group): 3-months-old group (young group); and 13-months-old group (aged group). After euthanasia, the head of the mandible was collected and stained with hematoxylin and eosin (HE) to evaluate the thickness of the articular layer and cartilage. Sections stained with Picrosirius red and Safranin O were used to evaluate the collagen and proteoglycans deposition, respectively. First, aging has decreased the articular layer thickness. Second, the results suggest a decrease of chondrocytes followed by an increase of the matrix to maintain the mandible homeostasis. Finally, both collagen and proteoglycans increased with aging. Aging displayed important effects to the mandible of aged rats.

    更新日期:2019-12-29
  • Impact of vildagliptin on vascular and fibrotic remodeling of myocardium in experimental diabetic cardiomyopathy
    Acta Histochem. (IF 1.719) Pub Date : 2019-12-28
    Ahmed A.M. Abdel-Hamid; Alaa El-Din L. Firgany

    The effect of dipeptidyl peptidase-4 inhibitors (DPP-4is) on myocardium in diabetic cardiomyopathy (DCM) remains a matter of debate. In the current study we investigated the effect of vildagliptin (VILDA, 3 mg/kg/d) on myocardium of DCM focusing on coronary microcirculation as well as on endothelial stress markers (ICAM and VCAM). We divided animals equally into 4 groups; nondiabetic (ND), VILDA per se, DCM and DCM + VILDA and their myocardium was evaluated for the fibro-vascular remodeling immunohistochemically as well as for molecular changes. VILDA had reversed the histological changes occurred in DCM including the disintegration, degeneration, and steatosis of cardiomyocytes with disappearance of the edema fluid. In addition VILDA significantly increased (p < 0.05) density of the coronary microcirculation and relieved endothelial stress. However, it did not prevent the development of fibrotic remodeling including the increased collagen deposition and the significantly upregulated (p < 0.05) corresponding genes. Therefore VILDA may have a positive impact on the microvascular remodeling, but not on fibrotic changes, in DCM.

    更新日期:2019-12-29
  • Could cathepsin-k be a driver of the myofibroblastic differentiation observed in dermatofibroma, atypical fibroxanthoma and pleomorphic dermal sarcoma?
    Acta Histochem. (IF 1.719) Pub Date : 2019-12-27
    Costantino Ricci; Antonio De Leo; Emi Dika; Martina Lambertini; Giulia Veronesi; Barbara Corti

    Dermatofibroma (BFH), atypical fibroxanthoma (AFX) and dermal pleomorphic sarcoma (DPS) are skin-based soft-tissue neoplasms of uncertain lineage. They are classified as “fibrohistiocytic” neoplasms, even if the World Health Organization stated that this term connotes a polymorphic group of lesions that histologically resemble fibroblasts and histiocytes. It is well-known that this group of lesions shows a “fibro-histiocytic-dendritic” and/or a “myofibroblastic” phenotype, even within the same lesion. We studied the expression of cathepsin-k in 34 cases (25 BFH, 5 AFX, 4 DPS) with a broad panel of antibodies. 20 cases (5 dermatofibrosarcoma protuberans, 5 melanomas, 5 basal cell carcinomas, 5 squamous cell carcinomas) were chosen as controls. Although our results need to be validated, they support a myofibroblastic and/or partial myofibroblastic (“proto-myofibroblastic”) phenotype and the lineage-plasticity of these neoplasms, highlighting the potential role of cathepsin-k in myofibroblastic trans-differentiation. Cathepsin-k proved to be an additional immunoistochemical marker potentially useful in the diagnostic algorithm.

    更新日期:2019-12-27
  • Effect of 1,25(OH)2-vitamin D3 on expression and phosphorylation of progesterone receptor in cultured endometrial stromal cells of patients with repeated implantation failure
    Acta Histochem. (IF 1.719) Pub Date : 2019-12-24
    Hossein Hosseinirad; Marefat Ghaffari Novin; Sedighe Hosseini; Hamid Nazarian; Fardin Amidi; Shahrokh Paktinat; Elham Azizi; Zahra Shams Mofarahe

    Repeated implantation failure (RIF) occurs in a condition when good quality embryos fail to implant in the endometrium following several in vitro fertilization (IVF) cycles. Suboptimal endometrial receptivity is one of the main underlying factors that causes this failure. Progesterone is the key regulator of endometrial receptivity which regulates gene expression through binding to its receptors in the endometrial stromal cells (eSC). The aim of this study was to evaluate the effect of 1,25(OH)2-vitamin D3 on progesterone receptor (PR) expression level and its phosphorylation on Ser294 residues in eSC of RIF patients and healthy fertile women. After isolation of the eSC from biopsy samples of RIF patients and healthy fertile women and their characterization, the cells were incubated with vitamin D3 and the expression level of PR mRNA, PR protein and phospho-Ser294 PR protein were evaluated after treatment. The results showed that vitamin D3 treatment increases PR mRNA and protein level and phospho-Ser294 PR protein level in the isolated eSC of both RIF patients and the control group. These results suggest that vitamin D3 may possibly play a key role during the embryo implantation process by affecting the expression pattern and regulatory modifications of the PR in the eSC.

    更新日期:2019-12-25
  • Prognostic value of connective tissue growth factor and c-Myb expression in IgA nephropathy and Henoch-Schönlein purpura—A pilot immunohistochemical study
    Acta Histochem. (IF 1.719) Pub Date : 2019-12-20
    Maja Mizdrak; Natalija Filipović; Katarina Vukojević; Vesna Čapkun; Ivan Mizdrak; Merica Glavina Durdov

    Aim Adverse and advanced prognostic signs in IgA nephropathy (IgAN) are interstitial fibrosis and tubular atrophy, but early predictors of bad outcome are still lacking. We investigated expression of connective tissue growth factor (CTGF) and c-Myb in renal biopsies of IgAN and Henoch-Schönlein purpura (HSP), because these gene products are indirectly included in fibrosis and epithelial-mesenchymal transition (EMT). Methods The sample included 23 patients and 8 controls who underwent nephrectomy due to renal cancer. The slides cut from the paraffin blocks were prepared for standard indirect immunoflourescence, using antibodies to CTGF and c-Myb. Ten high-power non-overlapping fields were photographed on Olympus IX51 microscope. Average percent of positive tubular cells, as well as number of positive cells per glomerulus were calculated. Results The cytoplasmic tubular CTGF expression was higher in IgAN/HSP than in controls (P < 0.001), whereas no difference was found in glomeruli (P = 0.437). The nuclear c-Myb expresssion in glomeruli and tubules was higher in IgAN/HSP than in controls (P < 0.05). In the follow-up, decline in renal function correlated with glomerular and tubular c-Myb, as well as tubular CTGF expression (all P < 0.05). Conclusion: Our results proposed c-Myb and CTGF as novel, early and sensitive markers of chronic kidney disease and worse renal outcome, but larger series are needed.

    更新日期:2019-12-21
  • High mobility group box 1 antibody represses autophagy and alleviates hippocampus damage in pilocarpine-induced mouse epilepsy model
    Acta Histochem. (IF 1.719) Pub Date : 2019-12-20
    Cui Ying; Liang Ying; Liu Yanxia; Wang Le; Cao Lili

    As a neurological disorder, epilepsy has affected over 65 million people all over the world because of the unforeseeable seizures it might cause. However, in-depth understandings of the pathogenesis of epilepsy and effective treatments for the disease are still lacked. Recent discoveries suggest that autophagy, as an endogenous self-cleansing pathway in mammals, might be involved in the onset of epilepsy. Our study assumes that a non-histone DNA binding protein, high mobility group box-1 (HMGB1), formerly considered as a crucial inflammatory factor, may mediate the autophagy of neurons in epileptic mouse brain. To verify this hypothesis, pilocarpine induced epilepsy mouse model was constructed. The mice were treated with HMGB1 antibody for 4 weeks after the initial epileptic seizure. Behavioral test results suggested a recovery of learning ability and memory in epileptic mice when treated with HMGB1 antibody. Pathological changes in hippocampus were inspected under microscopes and hippocampus damages caused by seizures in mouse with epilepsy such as increased intracellular space were alleviated by HMGB1 antibody treatment. Moreover, the expressions of the proteins involved in autophagy pathways were detected by immunofluorescence staining and western blot. microtubule-associated protein 1A/1B-light chain 3 (LC3), Beclin 1, autophagy protein-5 (ATG5), and ATG7 levels were significantly decreased by HMGB1 antibody while the level of p62 was increased. TdT-mediated dUTP Nick-End Labeling (TUNEL) illustrated that cell apoptosis induced by seizures in hippocampus was mitigated by HMGB1 antibody. In conclusion, we propose that HMGB1 may induce increased autophagy in epilepsy mouse model.

    更新日期:2019-12-20
  • The protective effect of curcumin on the diabetic rat kidney: A stereological, electron microscopic and immunohistochemical study
    Acta Histochem. (IF 1.719) Pub Date : 2019-12-18
    Mahmut Ulubay; Işınsu Alkan; Kıymet Kübra Yurt; Süleyman Kaplan

    The prevalence of diabetes in the world is increasing rapidly. Kidney diseases are among the most common medical disorders. The aim of this study was to investigate the effect of curcumin on the diabetic kidney. Thirty-five female Wistar albino rats were divided into seven groups. No procedure was performed on the Cont group. The Sham group received corn oil via gavage for 14 days. The curcumin (Curc) group received 30-mg/kg curcumin for 14 days, while the diabetes mellitus (DM) group received 50-mg/kg streptozotocin (STZ) in a single dose intraperitoneally. The DM + curcumin 1 (DC1) group received 30 mg/kg curcumin for 14 days, seven days after the application of STZ, while the DM + curcumin 2 (DC2) received 30 mg/kg curcumin for 14 days, 21 days after the application of STZ, and the DM + curcumin 3 (DC3) group received single-dose STZ at the same time as the application of 30 mg/kg curcumin for 14 days. Medulla, cortex, tubule, and glomerulus volume ratios were calculated using stereological techniques. Cortex volumes in the Sham and DM groups were significantly lower than in the Cont group (p < 0.05). The cortex volume in the DC3 group was also significantly lower than in the Curc group (p < 0.05). Medullary volume was significantly higher in the DC3 group compared to the DM group (p < 0.05). Curcumin was determined to exhibit a protective effect on the diabetic kidney since the glomerulus in the curcumin-exposed group exhibited a well-protected structure following experimentally induced diabetes based on light and electron microscopic analysis findings. These findings suggest that curcumin used following experimentally induced diabetes exhibits protective effects on the diabetic kidney.

    更新日期:2019-12-18
  • Differential mucins secretion by intestinal mucous cells of Chelon ramada in response to an enteric helminth Neoechinorhynchus agilis (Acanthocephala)
    Acta Histochem. (IF 1.719) Pub Date : 2019-12-18
    Giampaolo Bosi; Joseph A. DePasquale; Emanuele Rossetti; Bahram Sayyaf Dezfuli

    Intestinal mucous cells produce and secrete mucins which hydrate, lubricate and protect the intestinal epithelium from mechanical injuries due to the transition of digesta or action of pathogens. Intestinal mucous cells are considered elements of the innate immune system as they secrete lectins, toxins, immunoglobulins, and anti-microbial peptides. Acid mucins can surround and eliminate many pathogenic microorganisms. We performed a quantitative analysis of the density and mucus composition of different intestinal mucous cell types from mullet (Chelon ramada) that were infected solely with Neoechinorhynchus agilis. Most N. agilis were encountered in the middle region of the intestine. Mucous cell types were identified with Alcian Blue (pH2.5) and Periodic acid-Schiff (PAS) histochemistry, and by staining with a panel of seven lectins. Mucus enriched for high viscosity acid mucins was accumulated near points of worm attachment. Parasites were surrounded by an adherent mucus layer or blanket. Ultrastructural examination showed intestinal mucous cells typically possessed an elongated, basally positioned nucleus and numerous electron dense and lucent vesicles in the cytoplasm. The results show both an increase in mucus production and changes in mucin composition in infected mullet in comparison with uninfected conspecifics.

    更新日期:2019-12-18
  • The effects of magnesium sulfate on cyclophosphamide-induced ovarian damage: Folliculogenesis
    Acta Histochem. (IF 1.719) Pub Date : 2019-12-04
    Tuğba Ekiz Yılmaz, Müge Taşdemir, Mehmet Kaya, Nadir Arıcan, Bülent Ahıshalı

    Cyclophosphamide (CYP) is one of the alkylating chemotherapeutic agents and its adverse effects on folliculogenesis in the ovary are well-known due to the previous scientific research on this topic. Magnesium has various effects in organisms, including catalytic functions on the activation and inhibition of many enzymes, and regulatory functions on cell proliferation, cell cycle, and differentiation. In this study, the effects of magnesium sulfate (MgSO4) on CYP induced ovarian damage were investigated. Immature Wistar-Albino female rats of 28-days were treated with pregnant mare serum gonadotrophin (PMSG) to develop the first generation of preovulatory follicles. Rats of the experimental groups were then treated with either CYP (100 mg/kg, i.p) and MgSO4 (270 mg/kg loading dose; 27 mg/kg maintenance doseX12, i.p) solely or in combination. Following in-vivo 5-bromo-2-deoxyuridine (BrdU) labeling, animals were sacrificed and ovaries were embedded in paraffin and Epon. In the ovaries, added to the evaluation of general morphology and follicle count; BrdU and TUNEL-labeling, cleaved caspase-3 and p27 (cyclin-dependent kinase inhibitor) staining was also performed immunohistochemically and an ultrastructural evaluation was performed by transmission electron microscopy (TEM). The number of primordial follicles were decreased and multilaminar primary and atretic follicles were increased in CYP group. After MgSO4 treatment, while primordial follicle pool were elevated, the number of atretic follicles were decreased. Additionally, decreased BrdU-labeling, increased cleaved caspase 3 immunoreactivity and increased TUNEL labeling were observed in CYP group. In CYP treated animals, observations showed that while MgSO4 administration caused no alterations in BrdU proliferation index and caspase-3 immunoreactivity, it significantly reduced the TUNEL labeling. It was also observed that, while p27 immunoreactivity significantly increased in the nuclei of granulosa and theca cells in the CYP group; MgSO4 treatment significantly reduced these immunoreactivities. The ultrastructural observations showed frequent apoptotic profiles in granulosa and theca cells in both early and advanced stages of follicles in the CYP group and the MgSO4 treatment before the CYP application led to ultrastructural alleviation of the apoptotic process. In conclusion, our data suggest that MgSO4 may provide an option of pharmacologic treatment for fertility preservation owing to the beneficial effects of on chemotherapy-induced accelerated follicular apoptotic process, and the protection of the primordial follicle pool.

    更新日期:2019-12-04
  • Involvement of proliferative and apoptotic factors in the development of hindgut in rat fetuses with ethylenethiourea-induced anorectal malformations
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-28
    Caiyun Long, Yunxia Xiao, Siying Li, Xiaobing Tang, Zhengwei Yuan, Yuzuo Bai

    Background Anorectal malformations (ARMs) are common congenital malformations of the terminal digestive tract, but little is known regarding their pathogenesis. Aberrant cell proliferation/apoptosis are believed to be involved in ARMs. However, there are no studies on proliferation/apoptosis-related genes. Purpose We aimed to investigate the spatiotemporal expression patterns of two proliferation/apoptosis-related genes (MYC proto-oncogene and tumor protein p53) and explore their potential functions in the hindguts of ethylene thiourea-induced ARMs rat fetuses. Methods MYC and p53 expression was evaluated using immunohistochemical staining, western blotting, and quantitative real-time polymerase chain reaction (RT-qPCR). Terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) and p53 costaining were performed to assay the colocalization of apoptotic and p53-expressing cells. Results Rat fetuses with ARMs displayed fusion failure of the urogenital septum and cloacal membrane. In the control group, MYC was persistently expressed from gestational day (GD)14 to GD16 and distributed throughout the hindgut, while p53 was weakly detected in the terminal segment of the urethra and hindgut; in the ARMs group, MYC expression was obviously reduced, while p53 was widely and highly expressed in the urethra and hindgut. Western blotting and RT-qPCR confirmed the decrease in MYC and increase in p53 expression in ARMs. TUNEL and p53 co-staining revealed considerable overlap between apoptotic and p53-expressing cells. Conclusion The expression patterns of c-myc and p53 were disrupted in ARMs rat embryos, and the downregulation of c-myc and upregulation of p53 might be related to the development of ARMs at the key time points of ARMs morphogenesis.

    更新日期:2019-11-29
  • Expression of Langerin/CD207 in airways, lung and associated lymph nodes of a stranded striped dolphin (Stenella coeruleoalba)
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-27
    E.R. Lauriano, S. Pergolizzi, P. Lo Cascio, M. Kuciel, N. Zizzo, M.C. Guerrera, M. Aragona, G Capillo

    The airways and lungs of vertebrates are an entrance way for several microbial pathogens. Cetaceans present an upper and lower respiratory anatomy that allows the rapid flow of large air volumes, which may lead to high susceptibility to respiratory infections. Mortality and stranding rate of Cetaceans increased dramatically, so wide the knowledge about the immune system and specific antibodies identifying immune cells populations, is of fundamental importance to monitor and document cetacean health. The aim of this study was to identify the localization of dendritic cells marked by Langerin/CD207 in airways, lungs and associated lymph nodes, of the striped dolphin Stenella coeruleoalba. Samples of trachea, bronchi, lungs and lung-associated lymph nodes were obtained from a stranded adult male of Stenella coeruleoalba. Our results showed abundant lymphoid aggregates (LAs) in the lung of S. ceruleoalba. Langerhans-like dendritic cells were well distributed along the epithelium and interstitium of respiratory tract and in associated lymph nodes. The present study deepens the knowledge about the cetacean’s immune system and report about the exploitability of a commercial antibody (Langerin/CD207) for cetacean species.

    更新日期:2019-11-28
  • Inferring lanosterol functions in the female rabbit reproductive tract based on the immunolocalization of lanosterol 14-demethylase and farnesoid beta-receptor
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-26
    Maribel Méndez-Tepepa, Dafne Zepeda-Pérez, Leticia Nicolás-Toledo, Lourdes Arteaga-Casteñeda, Gabriel Gutiérrez-Ospina, Estela Cuevas-Romero

    Female reproductive organs have de novo synthesis of cholesterol. Some sterol molecules, intermediaries in the cholesterol synthesis, have important paracrine/autocrine actions. Lanosterol binds to the farnesoid beta-receptor (FXRβ), a molecule widely expressed in the ovaries, suggesting that it may play a role in reproduction. Up to date, we know little about lanosterol functions across female reproductive organs. We described immunolocalized lanosterol 14-demethylase (LDM or CYP51A1), responsible for catalyzing the conversion of lanosterol in cholesterol, and FXRβ in the ovary, oviduct, uterus, and vagina of virgin and pregnant rabbits. In virgin rats, we found CYP51A1 and FXRβ immunoreactivity was found in all ovarian follicles, epithelial cells, stroma, and Graafian follicles. Also, the epithelium and stroma, as well as the smooth muscle of the oviduct, vagina, and uterus showed CYP51A1 and FXRβ immunoreactivity. In pregnant dams, we observed the presence of CYP51A1 and FXRβ immunoreactivity in the corpora lutea, giant uterine cells, and trophoblastic cells. The presence of CYP51A1 and FXRβ support that lanosterol participates in diverse reproductive processes, including follicular maturation, transport of gametes and zygote, implantation of blastocyst, lubrication, and contraction of the vagina, secretion of female prostate, and control of delivery mediated by pelvic muscles contraction.

    更新日期:2019-11-27
  • Vesicular glutamate transporter 2-immunoreactive afferent nerve terminals in rat carotid sinus baroreceptors
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-26
    Takuya Yokoyama, Kazuya Settai, Nobuaki Nakamuta, Yoshio Yamamoto

    Sensory nerve endings respond to various stimuli and subsequently transmit afferent informations to central nervous system, but their responsibility has been suggested to be modulated by glutamate. In the present study, we examined the immunohistochemical localization of vesicular glutamate transporter 1 (vGLUT1) and vGLUT2 in baroreceptor nerve endings immunoreactive for P2X2 and P2X3 purinoceptors in the rat carotid sinus by immunohistochemistry of whole-mount preparations with confocal scanning laser microscopy. P2X3-immunoreactive flat leaf-like axon terminals were immunoreactive to vGLUT2, but not to vGLUT1. Among members of the soluble N-ethylmaleimide-sensitive factor attachment protein receptor complex, immunoreactivities for synaptosomal-associated protein, 25 kDa, Syntaxin1, and vesicle-associated membrane protein 2 (VAMP2) were localized in P2X2- and P2X3-immunoreactive axon terminals. Punctate immunoreactive products for VAMP2 and vGLUT2 were co-localized in axon terminals. These results suggest that vGLUT2 is localized in P2X3-immunoreactive baroreceptor terminals in the carotid sinus, and these terminals may release glutamate by exocytosis in order to modulate baroreceptor function in the carotid sinus.

    更新日期:2019-11-27
  • Fibroblast growth factor-2 signaling modulates matrix reorganization and cell cycle turnover rate in the regenerating tail of Hemidactylus flaviviridis
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-26
    Anusree Pillai, Sonam Patel, Isha Ranadive, Isha Desai, Suresh Balakrishnan

    Lizards restore their lost tail by the recruitment of multipotent cells which are selectively differentiated into varied cell types so as to sculpt a new tail. The precise coordination of the events involved in this complex process requires crosstalk between many signaling molecules and differential regulation of several mediators that facilitate the achievements of various milestones of regeneration. Fibroblast growth factor-2 is one such signaling molecule which activates a number of intracellular signaling pathways. Herein, the regulatory role of FGF2 during tail regeneration in Hemidactylus flaviviridis was investigated. Upon inhibition of FGFR using SU5402, the FGF2 levels were found to be significantly reduced at both transcript and protein level. Further, the compromised levels of the gelatinases, namely MMP2 and MMP9 in the tail tissues of treated lizards indicate that FGF2 regulates the activity of these enzymes perhaps to facilitate the recruitment of multipotent mesenchymal cells (blastema). The in vivo 5BrdU incorporation assay showed a lower cell proliferation rate in FGF2 signal inhibited animals during all the proliferative stages of regeneration studied. This observation was substantiated by decreased levels of PCNA in treated group. Moreover, from the combined results of Caspase-3 localization and its expression levels in the regenerates of control and SU5402 treated lizards it can be deduced that FGF2 signal regulates apoptosis as well during early stages of regeneration. Overall, the current study indicates beyond doubt that FGF2 signaling plays a pivotal role in orchestrating the matrix reorganization and cell cycle turnover during lizard tail regeneration.

    更新日期:2019-11-27
  • 更新日期:2019-11-27
  • Effects of uterus derived mesenchymal stem cells and their exosomes on asherman’s syndrome
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-24
    Gulistan Sanem Saribas, Candan Ozogul, Meral Tiryaki, Ferda Alpaslan Pinarli, Sevtap Hamdemir Kilic

    Asherman’s syndrome has become a growing problem with the incidence of cesarean and endometrial surgical procedures. A surgical procedure that can damage to the basal layer of the endometrium is formed as intrauterine adhesion and can cause asherman’s syndrome. Mesenchymal stem cells (MSCs) are characterized by some characteristics such as non-immunogenic, angiogenic, antifibrotic, antiapoptotic and antiinflammatory properties, also they support tissue repair by secretion of various factors and chemokines in cellular therapy. Exosomes are active paracrine components with a great potential for repairing damaged tissue. Exosomes include many paracrine factors responsible for regeneration and angiogenesis. In this study, 10 newborn Wistar rats were used to obtain MSCs. A total of 24 adult Wistar rats were also used. The rats were divided into 4 groups: untreated control group; asherman control group; asherman + uterine-derived MSCs group; asherman + uterine-derived MSCs-exosomes group. At the end of the experiment, uterine tissues were evaluated by histochemical and immunohistochemical. As a result of MSCs and exosomes treatments, proliferation and vascularization in uterine tissue was increased. It was also shown to reduce fibrosis with masson’s trichrome staining. MMP-2 and MMP-9 expression was enhanced by MSC and exosomal therapy; in addition, TIMP-2 expression was decreased. In our study, it was shown that proliferation and vascularization increased and fibrosis decreased in uterus as a result of MSC and exosome treatments. Our results indicate that the exosomal treatment restored the damage of asherman’s syndrome at tissue at a shorter time than the MSCs group.

    更新日期:2019-11-26
  • Pd-ligand 1 is expressed in inflammatory cells but not in neoplastic cells in hepatocellular carcinoma: An immunohistochemical study
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-22
    Francesco Vasuri, Azzurra Nerpiti, Stefano Zagnoni, Matteo Ravaioli, Antonia D’Errico, Michelangelo Fiorentino

    Nowadays, the major limit to the immunohistochemical (IHC) analysis of tissue PD-L1 is the high variability of the monoclonal antibodies commercially available. Aims of the present paper are to assess the best clone and the most suitable scoring for PD-L1 IHC determination on human hepatocellular carcinoma (HCC) among three commercially available clones, and to evaluate which PD-L1 clone is the best in predicting HCC aggressiveness in vivo. We built a tissue microarray (TMA) with 60 retrospective HCC cases, including the correspondent non-tumoral tissue. IHC was automatically performed using the following anti-PD-L1 clones: 28.8, SP142, and SP263. As results, we did not find any immunoreactivity for PD-L1 in both neoplastic and normal hepatocytes included in the TMA using the three antibodies. Positivity for PD-L1 was exclusively seen in inflammatory cells within the HCC tissue and in cirrhotic parenchyma. When a gold standard was assessed, the sensitivity of SP142, 28.8 and SP263 was 46 %, 54 % and 85 % respectively. Using the SP263 clone, the absolute number of PD-L1-positive inflammatory cells in the HCC cores was paired with the number of PD-L1-positive inflammatory cells in the corresponding non-tumoral tissue (P = 0.001). Finally, using SP263, the mean number of PD-L1-positive cells was 11.3 ± 12.6 in HCC from deceased patients, versus 4.7 ± 5.2 in alive patients (p = 0.039). SP263 is the most sensitive clone for PD-L1 IHC tissue determination in HCC, as well as the best antibody for the assessment of its biological behavior.

    更新日期:2019-11-22
  • Clenbuterol accelerates recovery after immobilization-induced atrophy of rat hindlimb muscle
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-21
    Hideki Suzuki, Yuki Yoshikawa, Hisaya Tsujimoto, Takashi Kitaura, Isao Muraoka

    Using immunohistochemistry, we investigated whether daily administration of clenbuterol (CLE; 1 mg/kg body weight per day) accelerates recovery after casted immobilization(IMM)-induced atrophy of fast-twitch plantaris and slow-twitch soleus muscles. Adult male Sprague-Dawley rats were assigned to the control (CON), casted immobilization (IMM), casted immobilization following recovery control (RCON), and casted immobilization following recovery with CLE administration (RCLE) groups. Casted immobilization and recovery periods were 9 and 14days, respectively. Rats of the CON group were subjected to the experiment simultaneously with the IMM group. Nine days of immobilization induced muscle fiber atrophy, which was greater in the soleus muscle than in the plantaris muscle. After the 2-week recovery period, the cross-sectional areas of each fiber type in both muscles were higher in the RCON group than in the IMM group. The cross-sectional areas of each fiber type in both muscles in the RCLE group were larger than those in the RCON group. The myonuclear number of each fiber type of the plantaris muscle in the RCON and RCLE groups was higher than that in the CON group. In contrast, the myonuclear number per fiber of the soleus muscle was not affected by hindlimb immobilization, reloading, and clenbuterol administration regardless of muscle fiber type. These results suggest that CLE accelerates the recovery of atrophied plantaris and soleus muscles fibers and that their mechanisms of responses to CLE in both muscles may be different during recovery period after muscle atrophy.

    更新日期:2019-11-22
  • Comparative analysis of fresh chondrocytes, cultured chondrocytes and chondroprogenitors derived from human articular cartilage
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-13
    Elizabeth Vinod, Upasana Kachroo, Soosai Manickam Amirtham, Boopalan Ramasamy, Solomon Sathishkumar
    更新日期:2019-11-13
  • In vitro effects of PNP and PNMC on apoptosis and proliferation in the hen ovarian stroma and prehierarchal follicles
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-08
    A.K. Grzegorzewska, A. Hrabia, K. Kowalik, D. Katarzyńska-Banasik, A. Kozubek, A. Sechman

    This study aimed to examine the mRNA expression, activity, and immunolocalisation of apoptosis/proliferation regulating factors following in vitro exposure of the stroma, white (WFs), and yellowish (YFs) follicles of the chicken ovary to 4-nitrophenol (PNP) or 3-methyl-4-nitrophenol (PNMC). PNMC increased the mRNA expression of caspase-3, -8, Apaf-1, and cytochrome c in the ovarian stroma. The activity of caspase-3, -8, and -9 decreased in WFs in both nitrophenol-treated groups. PNP reduced the number of caspase-3-positive cells in the stromal connective tissue (CT) and the theca interna and externa layers of WFs. In the stroma, the proliferating index decreased in the wall of primary follicles in both nitrophenol-treated groups, however, in the CT, the effect of PNMC was opposite. In the theca interna of WFs, PNP diminished the proliferating index. These results suggest that nitrophenols might impact the development of chicken ovarian follicles by affecting cell death and proliferation.

    更新日期:2019-11-08
  • Presence of serotonin and its receptor in the central nervous system and ovary and molecular cloning of the novel crab serotonin receptor of the blue swimming crab, Portunus pelagicus
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-07
    Jirasuda Nakeim, Napamanee Kornthong, Jirawat Saetan, Supawadee Duangprom, Prasert Sobhon, Prapee Sretarugsa

    Serotonin (5-HT) plays pivotal roles in many physiological processes including reproduction of crustaceans, which are mediated 5-HT receptors. The distributions of 5-HT and its receptor have never been explored in Portunus pelagicus. To validate the targets which indirectly indicate the roles of 5-HT in this crab, we have investigated the distribution of 5-HT in the central nervous system (CNS) and ovary using immunohistochemistry and tissue expression of its receptor by RT-PCR. In the brain, 5-HT immunoreactivity (-ir) was detected in clusters 6, 7, 8, 11, 14, 15 and the fibers. In the ventral nerve cord (VNC), 5-HT-ir was detected in pairs of neurons and the fibers connected to the neurons. In the ovary, 5-HT-ir was intense in the oocyte step 1 (Oc1) and Oc2, and its intensity was slightly decreased in Oc3 and Oc4. The 5-HT receptor was molecularly characterized to be type 7, and it was strongly expressed in the eyestalk, brain, VNC, mature ovary and muscle. Due to the presence of 5-HT receptor we suggest that 5-HT acts primarily at the CNS and ovary, thus implicating its role in reproduction especially in the development of oocytes though its exact function in this crab needed to be explored further.

    更新日期:2019-11-07
  • Prophylactic supplementation of 20-HETE ameliorates hypoxia/reoxygenation injury in pulmonary vascular endothelial cells by inhibiting apoptosis
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-06
    Praveenkumar Sugumaran, Vishnusekar Narayanan, Daling Zhu, Meetha Medhora, Elizabeth R. Jacobs, Yamini Chandramohan, Vimalraj Selvaraj, Anuradha Dhanasekaran

    Hypoxia reoxygenation (HR) injury perturbs structural and functional syncytium in lung tissues. It is commonly implicated in conditions such as stroke, lung transplant or severe pneumonia. In the present study, we investigated the cytoprotective action of 20-hydroxyeicosatetraenoic acid (20-HETE) on pulmonary vascular endothelial cells (PMVECs) under normoxic and hypoxic niche followed by HR. 20-HETE pretreatment showed a protective effect at a concentration of 1μM as there was a marked increase (20%) in the cell viability compared to control and HR groups. Pretreatment of 20-HETE in HR induced injury decreased ROS production dictated its antioxidant property. Similarly, SOD and ATP levels were also downregulated by 20-HETE pretreatment. Cell apoptosis was detected by TUNEL assay, Acridine orange, and procaspase-3 cleavage, caspase-3 activity assay, respectively. JC-1 mitochondrial membrane potential assay and protein expression pattern of BCL-2, and BAD phosphorylation status were examined. The results showed that HR induced significant increase of apoptotic PMVECs, while 20-HETE pretreatment attenuated the effects. Further, 20-HETE pretreatment activated PI3K/Akt and HIF-1α signaling pathway to exhibit its protective effects against HR-induced oxidative stress and apoptosis. Overall, the results concluded the potent antioxidant role of 20-HETE in aiding cytoprotection upon HR injury.

    更新日期:2019-11-06
  • Oral variant of acantholytic squamous cell carcinoma—Histochemical and immunohistochemical features
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-06
    Irit Allon, Michael Abba, Ilana Kaplan, Alejandro Livoff, Amram Zaguri, Oded Nahlieli, Marilena Vered

    Acantholytic squamous cell carcinoma (ASCC) is an uncommon variant of squamous cell carcinoma (SCC). It is characterized by a combination of typical SCC and pseudoglandular structures, dyskeratotic cells and prominent acantholysis. The purpose of this study was to analyze the histochemical and immunohistochemical characteristics of the intraoral variant of ASCC. Cases of intraoral ASCC were retrieved from the English language literature. Four new cases from our files were added. In total, 35 cases were included and analyzed in this study. The mean age of the patients was 61.5 + 13 years (age range 38–92 years), with a male-to-female ratio of 1.7:1. According to the available data, histochemical and immunohistochemical stains for mucins were found to be consistently negative. E- cadherin, a marker of adherens junctions, was usually reported to be expressed in areas of “typical” (non acantholytic) SCC, but reduced in the acantholytic areas. We examined for the first time the expression of claudin 1, a marker of tight junctions, and found it to be reduced in the acantholytic areas, similar to E-cadherin. Several cases of oral ASCC also expressed vimentin and cytokeratin (CK) 19, markers associated with epithelial-mesenchymal transition. A wide range of non-epithelial markers yielded negative immunoreactions. In conclusion, ASCC is an uncommon variant of squamous cell carcinoma. The acantholytic process appears to involve reduced expression of molecular components of both adherens junctions and tight junctions. These findings could suggest a relation to the epithelial mesenchymal transition process and therefore further studies are needed in order to establish such a link and the subsequent possible impact on the clinical outcome of the patients.

    更新日期:2019-11-06
  • Presence of metalloproteinases 2 and 9 and 8-OHdG in the fibrotic process in skeletal muscle of Mdx mice
    Acta Histochem. (IF 1.719) Pub Date : 2019-11-04
    Lidiane Begalli de Souza, Carla Maziero, Mariana Cruz Lazzarin, Hananiah Tardivo Quintana, Tabata de Carvalho Tomé, Vivianne Izabelle de Araújo Baptista, Flavia de Oliveira

    Inflammation and oxidative stress occurs in muscle of Duchenne muscular dystrophy (DMD). The relationship between a panel of biomarkers and the DMD outcome is necessary to indicate of disease progression and response to rehabilitation programs. The aim was to analyze the connective tissue of muscle of Mdx mice and immunoexpression of MMP-2, MMP-9, and 8-OHdG, which signalizes oxidative stress related to DNA damage. Biceps brachii of male C57BL/10 and C57BL/10-Dmdmdx mice was submitted to Hematoxylin-Eosin, Sirius red and immunohistochemistry (MMP-2, MMP-9 and 8-OHdG) analysis. Mdx showed focal lesions with intense inflammation and fibrosis related to immunoexpression of MMP-2 and MMP-9, proving the hypothesis that these MMPs are linked to muscular tissue degeneration, which can be regenerated by their inhibition, improving the treatment of DMD carriers. Histopathological findings related to centralized nuclei increase were related to higher 8-OHdG immunomarked nuclei in Mdx, which signalizes oxidative stress associated with DNA damage provoked by DMD. Such result shows that the evaluation of 8-OHdG during the evolution of the disease could be a method to evaluate DMD disease progression.

    更新日期:2019-11-04
  • Editorial.
    Acta Histochem. (IF 1.719) Pub Date : 2019-10-02
    Marilena Vered

    更新日期:2019-11-01
  • The dynamics of DAXX protein distribution in the nucleus of mouse early embryos.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-29
    Irina O Bogolyubova,Zhuldyz K Sailau,Dmitry S Bogolyubov

    Nuclear distribution of Death-associated protein 6 (Daxx) was studied using fluorescent and electron microscopy in mouse embryos at different stages of development in vivo, from zygote to morula. Daxx was found in association with transcriptionally silent chromatin predominantly with a heterochromatin rim surrounding the nucleolus precursor bodies (NPBs) at all stages studied. At the zygote stage, Daxx was detected only at the periphery of NPBs both in male and female pronuclei. At the late two-cell stage, Daxx was localized not only in the heterochromatin rim at the periphery of NPBs but also in heterochromatin zones not associated with NPBs. At the morula stage, a diffuse distribution of Daxx prevailed. Scarce Daxx-positive zones were detected only in some embryos at the nucleolar periphery. Thus, Daxx is noticeably redistributed during mouse embryo cleavage, and the most conspicuous areas of Daxx concentration are observed at the end of two-cell stage. Daxx is found colocalized with the chromatin-remodeling protein ATRX exclusively in two-cell embryos, but the heterochromatin areas containing either Daxx or ATRX individually are also observed at this stage. However, most zones containing both Daxx and ATRX demonstrated a low FRET-efficiency. This suggest that two molecules are not approached sufficiently close for molecular interactions to occur. Our data suggests that Daxx may function without cooperation with ATRX at least at some stages of early mouse development.

    更新日期:2019-11-01
  • Etmopterus spinax, the velvet belly lanternshark, does not use bacterial luminescence.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-28
    Laurent Duchatelet,Jérôme Delroisse,Patrick Flammang,Jacques Mahillon,Jérôme Mallefet

    Marine organisms are able to produce light using either their own luminous system, called intrinsic bioluminescence, or symbiotic luminous bacteria, called extrinsic bioluminescence. Among bioluminescent vertebrates, Osteichthyes are known to harbor both types of bioluminescence, while no study has so far addressed the potential use of intrinsic/extrinsic luminescence in elasmobranchs. In sharks, two families are known to emit light: Etmopteridae and Dalatiidae. The deep-sea bioluminescent Etmopteridae, Etmopterus spinax, has received a particular interest over the past fifteen years and its bioluminescence control was investigated in depth. However, the nature of the shark luminous system still remains enigmatic. The present work was undertaken to assess whether the light of this shark species originates from a bioluminescent bacterial symbiosis. Using fluorescent in situ hybridization (FISH) and transmission electron microscopy (TEM) image analyses, this study supports the conclusion that the bioluminescence in the deep-sea lanternshark, Etmopterus spinax, is not of bacterial origin.

    更新日期:2019-11-01
  • Increased mast cell number is associated with a decrease in beta-cell mass and regeneration in type 2 diabetic rats.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-25
    Ahmed A M Abdel-Hamid,Alaa El-Din L Firgany

    The role of mast cells (MCs) in type 2 diabetes (T2D) is not thoroughly studied as much as in T1D. Therefore in the current study we investigated correlation between these cells and various parameters of islets of Langerhans (IOL) in rats which were equally divided (n = 40) into; control and diabetic groups. We detected a significantly increased (p < 0.05) MC count (MCC) in the diabetic IOL compared to the control, together with a noticeable intra-islet seeding of these cells which displayed a tryptase positive immunostaining. A significant positive correlation (p < 0.05) between MCC and the % of glucagon cells per islet was detected in DM, unlike mass of the islets, mass of β-cells, and % of β-cells per islet which were negatively correlated with MCC. Similarly, there was a negative correlation between MCC with β-cell proliferation and neogenesis frequency in DM. This highlights the potential association between the increased MC number and the diminished islet`s mass as well as regeneration which may fasten the progression of T2D.

    更新日期:2019-11-01
  • Pre-administration of luteoline attenuates neonatal sevoflurane-induced neurotoxicity in mice.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-23
    Yang Wang,Chun Wang,Yue Zhang,Guan Wang,Haitao Yang

    Sevoflurane is a widely used inhaled anesthetic, which triggers neuroapoptosis and oxidative damage in the developing central nervous system and cognitive dysfunction later in life. However, no effective therapeutic strategy for sevoflurane-induced deleterious effects is well developed. The purpose of the present study was to explore whether luteoline could attenuate neonatal sevoflurane exposure-triggered neurotoxicity. In this study, six-day-old C57BL/6 mice were pretreated with luteoline (30, 60 mg/kg) intraperitoneally for 30 min before exposed to 3% sevoflurane 6 h consecutively. We first examined the effects of luteoline on hippocampal neuron apoptosis, inflammation and oxidative stress 18 h post anesthesia. The spatial learning and memory performance was measured using Morris water maze test from postnatal day 31 to 38. The results showed that luteoline ameliorated neuronal apoptosis as evidenced by decrease of apoptotic cells, downregulation of the cleavage levels of caspase-3 and PRAP, and inactivation of caspase-3. Moreover, luteoline significantly decreased protein expressions of inflammatory cytokines (IL-1β, IL-18 and TNF-α), inhibited NF-кB/NLRP3 pathway (NF-кB, NLRP3, ASC and caspase-1) and suppressed NF-кB activity. Our analyses indicated that luteoline had a significant effect on decreasing the contents of ROS and MDA, elevating the activity of SOD, and ultimately improving spatial learning and memory deficits of mice. In summary, our findings confirm that the attenuation of luteoline on sevoflurane-induced spatial learning and memory impairment later is associated with inhibition of hippocampal neuron apoptosis, inflammation and oxidative stress early. Luteoline might be a potential therapeutic for sevoflurane anesthesia-induced neurobehavioral dysfunction.

    更新日期:2019-11-01
  • Survivability of rabbit amniotic fluid-derived mesenchymal stem cells post slow-freezing or vitrification.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-22
    Barbora Kulikova,Michal Kovac,Miroslav Bauer,Maria Tomkova,Lucia Olexikova,Jaromir Vasicek,Andrej Balazi,Alexander V Makarevich,Peter Chrenek

    This work aimed to evaluate the effect of two distinct cryopreservation procedures - conventional slow-freezing and vitrification, on survivability and mesenchymal marker expression stability of rabbit amniotic fluid-derived mesenchymal stem cells (rAF-MSCs). Cells at passage 2 were slowly frozen, using 10% of dimethylsulfoxide, or vitrified, using 40% of ethylene glycol, 0.5 M sucrose and 18% Ficoll 70. After three months storage in liquid nitrogen, viability, chromosomal stability, ultrastructure, surface and intracellular marker expression and differentiation potential of cells were evaluated immediately post-thawing/warming and after additional culture for 48-72 h. Our results showed decreased (P ≤ 0.05) viability of cells post-thawing/warming. However, after additional culture, the viability was similar to those in fresh counterparts in both cryopreserved groups. Increase (P ≤ 0.05) in the population doubling time of vitrified cells was observed, while doubling time of slow-frozen cells remained similar to non-cryopreserved cells. No changes in karyotype (chromosomal numbers) were observed in frozen/vitrified AF-MSCs, and histological staining confirmed similar differentiation potential of fresh and frozen/vitrified cells. Analysis of mesenchymal marker expression by qPCR showed that both cryopreservation approaches significantly affected expression of CD73 and CD90 surface markers. These changes were not detected using flow cytometry. In summary, the conventional slow-freezing and vitrification are reliable and effective approaches for the cryopreservation of rabbit AF-MSCs. Nevertheless, our study confirmed affected expression of some mesenchymal markers following cryopreservation.

    更新日期:2019-11-01
  • Localization of phospholipase C β3 in the major salivary glands of adult mice.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-21
    Atsara Rawangwong,Atthapon Pidsaya,Wipawee Thoungseabyoun,Apussara Tachow,Tarinee Sawatpanich,Waraporn Sakaew,Miwako Yamasaki,Masahiko Watanabe,Hisatake Kondo,Wiphawi Hipkaeo

    Phospholipase C (PLC)β has a role in saliva secretion by controlling intracellular Ca2+via its product, IP3. The present study was attempted to localize PLCβ isoforms in mouse salivary glands in situ. A single major band was detected for PLCβ3 in immunoblots of the parotid and sublingual glands (PG, SLG), while no such band was seen in the submandibular gland (SMG). No bands were detected for PLCβ1 or 4 in the three glands. In immuno-light microscopy of PG and SLG, substantial immunoreactivity for PLCβ3 was seen in the cytoplasm including the plasmalemma of almost all ductal cells, while no distinct immunoreactivity was discerned in most acinar cells except for sublingual demilune cells. Numerous ductal cells exhibited higher immunoreactivity for PLCβ3 in their apical/supranuclear cell domain including the plasmalemma than in the basal/infranuclear domain, indicating an apico-basal polarity. In immuno-gold electron microscopy of PG ducts and SLG ducts and demilunes, most gold particles were found in association with plasma membranes as well as various intracellular membranes, most of which formed small oblong or flattened vesicles and vacuoles. A few particles were seen without association with any membranous structures. The present finding supports the previous physio-pharmacological result that Ca2+-signaling proteins as well as initial intracellular Ca2+ changes occur in the apical cell domain including the plasma membranes of the exocrine cells.

    更新日期:2019-11-01
  • Characterization of fat body cells at different developmental stages of Culex pipiens.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-14
    Gamze Turgay-İzzetoğlu,Mehtap Gülmez

    The fat body, originates from mesoderm, has many metabolic functions which changes as the embryonic development of the insect progresses. It plays an important role in the intermediate metabolism and in the metabolism of proteins, lipids and carbohydrates. It has roles in synthesis, absorption and storage of nutrients from hemolymph. It is also responsible for the production of immunological system components, antibacterial compounds and blood clotting proteins. The most common type of fat body cells are trophocytes (the basic cells of the fat body) and oenocytes are found associated with the fat body. In this study, it is aimed at determining the cell types contained in the fat body of Culex pipiens at different developmental stages as well as identifying the molecules such as carbohydrate, protein and lipid contained in each of these cells. Knowing the regional distribution of the fat body cells and the concentration of its content at each developmental stage is important in understanding the process related to its physiology and it may help in fighting against the pest C. pipiens, which is a vector species for many contagious diseases observed in humans and other species. To achieve our goal, we have employed different histochemical techniques (fixatives and staining methods) for staining C. pipiens preparates of different developmental stages and analyzed the structure of the fat body, its distribution, its cell types and the macromolecular contents of the cells. We only observed trophocytes and oenocytes as fat body components in C. pipiens. The trophocytes had all the three macromolecules (lipids, proteins, carbohydrates) in the cytoplasm varying in concentration between the different regions and different stages. The oenocytes were observed below the integument as well as between the muscles in the larvae of Culex pipiens. They were present either as single cells or in clusters and also varied in size. Their cytoplasm was stained strongly for proteins when bromophenol blue staining was applied, but it was rather heterogeneous due to the lipid inclusions. On the contrary, oenocytes were not observed among the adult C. pipiens preparations.

    更新日期:2019-11-01
  • The possible alleviating effect of saffron on chlorpyrifos experimentally induced cardiotoxicity: Histological, immunohistochemical and biochemical study.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-13
    Hanaa Attia Khalaf,Ahmed Abd El-Rahman El-Mansy

    INTRODUCTION Pesticides are responsible for many occupational health hazards among farmers in developing countries. Chlorpyrifos (CPF) is one of the broad-spectrum organophosphorus (OP) insecticides used for agricultural, domestic and industrial purposes. AIM OF THE WORK The present study was designed to examine the effects of CPF on cardiac muscles and to evaluate the possible protective role of crocin using biochemical and histological methods with the intention to recognize the molecular tools of its probable cardioprotective effects. MATERIALS AND METHODS Thirty-six adult male albino rats were used in this study and were divided into 4 equal groups (9 rats each): negative control group, positive control group, CPF treated group and CPF & crocin treated group. The heart was removed for histological and immunohistochemical studies. RESULTS Stained sections of cardiac muscle fibers of group III with H&E revealed remarkable histological changes in the form of disorganization of the fibers with increase in the interstitial spaces between these fibers. Congested dilated blood capillaries could be observed with extravasation of the red blood cells leading to interstitial hemorrhage. Focal areas of mononuclear cellular infiltration could be seen in the interstitial tissue. A number of cardiac fibers achieved pale acidophilic vacuolated sarcoplasm while others achieved dark homogenous acidophilic sarcoplasm. Some nuclei were peripherally situated and pyknotic while others were centrally situated and encircled with halos. Apparently increased masses of collagen fibers among the cardiac muscle fibers and around the congested dilated blood vessels with the presence of focal parts of extensive collagen fiber deposition were noticed in Mallory-stained sections of group III. Strong positive immunoreactions in the endomysium and perimysium of the cardiac fibers, along with the walls of blood capillaries and in interstitial cells, could be detected in immunohistochemical staining sections of group III with vimentin antibody. Immunoreactivity to caspase 3 was higher in the sarcoplasm of the cardiac fibers of group III compared to that of control group. A highly significant decrease in the cardiac level of SOD and CAT; however, a highly significant increase in MDA level was noted between the control groups and CPF treated group. Additionally, there was a significant improvement of the chemical and histological representations of group IV, and these improvement pictures were toward the normal. CONCLUSION The study concludes that crocin can alleviate the toxic effect of chlorpyrifos caused by oxidative stress on cardiac muscle.

    更新日期:2019-11-01
  • Beta-hydroxybutyric acid attenuates neuronal damage in epileptic mice.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-08
    Qinghui Guo,Shangming Liu,Shandan Wang,Meiyan Wu,Zhaoxia Li,Yibiao Wang

    β-Hydroxybutyric acid (BHBA) reportedly has neuroprotective and anti-oxidation properties. The present study aimed to investigate the protective effects of BHBA against epilepsy. C57BL/6 J mice were exposed to lithium chloride and pilocarpine to induce epilepsy and then were administrated with 300 mg/kg/day BHBA for 30 days. The learning impairment was evaluated via Morris Water Maze. Neuron loss and cell apoptosis were detected through Nissl staining and TUNEL staining. The levels of oxidative stress-related factors were determined by commercial kits. The protein expression levels of AMP-activated protein kinase (AMPK), p-AMPK, peroxisome proliferator-activated receptor alpha (PPARα), anti-apoptotic Bcl-2, and pro-apoptotic Bax were measured through Western blots. It was found BHBA improved epilepsy- caused learning deficiency and attenuated epilepsy-mediated neuron loss and cell apoptosis in the hippocampus. BHBA ameliorated oxidative stress via decreasing the levels of reactive oxygen species and malondialdehyde plus strengthening the activities of glutathione peroxidase and superoxide dismutase. BHBA also promoted the phosphorylation of AMPK and upregulated PPARα in the epileptic hippocampus. In conclusion, BHBA attenuates neuronal damage in epileptic mice, which is associated with its anti-apoptotic and anti-oxidative effects as well as the activation of AMPK and PPARα.

    更新日期:2019-11-01
  • An improved acridine orange staining of DNA/RNA.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-08
    Danilo M Damas-Souza,Rony Nunes,Hernandes F Carvalho

    New tools are desirable to examine the metabolic state of individual cells within tissues. We proposed a fluorescence-based procedure consisting of acridine orange staining and fast green counterstaining (AO-FG) to improve the selectivity of the former for nucleic acids (acridine orange stains both DNA and RNA with different fluorescence colors), with no interference from proteins. We compared this test with the biochemical quantification of the relative amounts of RNA and DNA in selected rat ventral prostate samples and PC3 cells. The epithelium of the prostate gland is highly active metabolically for the production of secretions. Differences in AO-RNA staining were revealed and correlated with the metabolic state of the epithelium. Specificity was confirmed by RNase A. To assess how AO-FG staining correlates with the metabolic state of the cell, we cultured PC3 cells in different concentrations of glucose and measured the ratios between the amounts of RNA and DNA. In parallel, similar cultures were subjected to AO-FG, and the staining pattern correlated closely (r2=0.886) with the obtained biochemical results. The results confirmed that the combined use of AO and FG is useful for detecting DNA and RNA simultaneously, as well as for assessing quantitatively the transcriptional activity of individual cells and their changes in response to experimental manipulation.

    更新日期:2019-11-01
  • Structural adequacy of the digestive tract supports dual feeding habit in catfish Pachypterus khavalchor (Siluriformes: Horabagridae).
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-07
    Sachin M Gosavi,Chandani R Verma,Sanjay S Kharat,Manoj Pise,Pradeep Kumkar

    Lepidophagy is comparatively rare amongst teleost fishes, yet our understanding of this specialization is lacking. Therefore we examined the digestive tract features of Pachypterus khavalchor using morphological, osteological, histological and histochemical techniques to comprehend and relate structural organization of digestive tract with scale eating habit. Morphologically, the alimentary canal is defined by a short and muscular esophagus, well-developed stomach and comparatively short intestine. Gut content analysis and intestinal coefficient value (0.53 ± 0.01) revealed that P. khavalchor exhibit both carnivory and lepidophagy. However, P. khavalchor primarily feeds on the scales (67.47%) and other chitin-rich material like aquatic insects (17.62%), aquatic larvae (8.66%) which affirms its solid association with chitinase producing endosymbionts in the gut. Lepidophagy is further supported by the osteological observations. The perfect segregation of the functions such as food capture, ingestion and processing amongst the different types of teeth located in the oral cavity and pharyngeal region thus could be taken as evolutionary adaptations in scale eaters to support lepidophagy. Specialized arrangement of the esophageal and stomach epithelial folds could be altogether taken as an adaptation with the end goal to frame the scale stacks and accordingly facilitate the handling and processing of chitin-rich bolus. The esophageal mucosa is simple squamous epithelium instead of stratified epithelium with numerous goblet cells to withstand the mechanical harm by hard-food stuff like scales. The cardiac and fundic regions exhibited large number tubular gastric glands with simple columnar epithelium. Surface cells of all three stomach regions stained positive for PAS staining. The intestine is without pyloric caeca and is divided into anterior and posterior region. Histologically it is characterized by simple columnar epithelium with brush border and numerous goblet cells throughout its length. Presence of large number microvilli on anterior and posterior intestine was noticeable. Intestinal goblet cells reacted positively to PAS, AB (pH 1) and AB (pH 2.5). Secretions of goblet cells are important for lubricating and protecting the epithelium. The results of present investigation improve the understanding of the digestive physiology of scale eaters in general and P. khavalchor in particular. Overall, our data indicates that though P. khavalchor predominantly feeds on scale, the digestive physiology is adapted to support dual feeding habit (lepidophagy and carnivory).

    更新日期:2019-11-01
  • Stereological examination of curcumin's effects on hippocampal damage caused by the anti-epileptic drugs phenobarbital and valproic acid in the developing rat brain.
    Acta Histochem. (IF 1.719) Pub Date : 2019-04-06
    Burcu Demirel Yılmaz,Banu Eren,Dilek Sağır,Zafer Eren,Ayşe Başardı Gökçe

    The anti-epileptic drugs phenobarbital and valproic acid have an extremely strong negative effect on cognitive processes such as learning and memory in the developing brain. We examined whether or not curcumin has protective effects on neuronal injury caused by these drugs in the developing rat brain. Young male Wistar rats were studied in two groups, a 7 days old and a 14 days old group (35 rats in each). Both groups were then divided into 7 sub-groups as the control, curcumin, dimethylsulfoxide, phenobarbital, valproic acid, phenobarbital + curcumin, and valproic acid + curcumin groups (n = 5 in each group). At 24 h after the intraperitoneal injection of the compounds, the rats were sacrificed, and the hippocampal tissue was subjected to stereological analysis with the optical fractionation method. Total numbers of neurons in the hippocampus of the 7 days old and 14 days old rats were calculated. It was found that treatment with phenobarbital resulted in a loss of 43% of the neurons, and valproic acid induced a loss of 57% of the neurons in the 7 days old rats. Curcumin prevented this loss significantly with only 19% in the phenobarbital group and 41% in the valproic acid group. In the 14 days old rat groups, phenobarbital was found to reduce the number of neurons by 30%, and valproic acid reduced it by 38%. Curcumin treatment limited neuronal loss to 3% in the phenobarbital + curcumin group and 10% in the valproic acid + curcumin group. These data strongly indicate that curcumin is a protective agent and prevents hippocampal neuronal damage induced by phenobarbital and valproic acid treatment.

    更新日期:2019-11-01
  • Basement membranes in the kidney of the dromedary camel (Camelus dromedarius): An immunohistochemical and ultrastructural study.
    Acta Histochem. (IF 1.719) Pub Date : 2019-03-25
    Lemiaa Eissa,Haider I Ismail,Mortada M O Elhassan,Hassan A Ali

    Basement membranes consist of various proteins, the major ones being laminin and collagen type IV. Primary defects in these two proteins have been extensively associated with kidney pathologies. This study aimed to establish baseline information on the immunohistochemical distribution of laminin and collagen type IV, and to corelate these with the ultrastructure of basal laminae in the uriniferous tubules of the dromedary camel. Tissue samples were taken from the kidneys of eight adult female camels, and processed for immunohistochemical and ultrastructural investigations. Strong intensity of collagen type IV was observed within the basement membranes of Bowman's capsule. The thickness of the basal lamina of the parietal layer of Bowman's capsule varied extensively depending on the region of the renal corpuscle; the thicker areas were always associated with cuboidal epithelial cells. The glomerular basement membrane revealed strong immunostaining of laminin, whereas the mesangial matrix was strongly immunoreactive to collagen type IV. Abundant amount of laminin was found in the basement membranes of proximal convoluted tubules, thin limbs of the loop of Henle, and collecting ducts. Dense immunostainings of laminin and collagen type IV were observed in the medullary regions of uriniferous tubule, in which numerous projections extended from the basal laminae into the subjacent connective tissue. Overall, the present study revealed marked variations in the distribution of the basement membrane markers laminin and collagen type IV in the uriniferous tubules of camel kidney. The results have also shown difference in the thickness of basal laminae. This variation in thickness, however, was unlikely to be influenced by the amount of laminin and collagen type IV.

    更新日期:2019-11-01
  • Epidermal growth factor receptor 2 immunoexpression in gastric cells of domestic cats with H. heilmannii infection.
    Acta Histochem. (IF 1.719) Pub Date : 2019-03-21
    Daniela Araujo de Sousa,Kássia Valéria Gomes Coelho da Silva,Clarice Marante Cascon,Franciele Basso Fernandes Silva,Marcela Freire Valim de Mello,Juliana da Silva Leite,Ana Beatriz Monteiro Fonseca,Kênia Balbi El-Jaick,Ana Maria Reis Ferreira

    The aim of this study was to evaluate the immunoexpression of HER-2 in gastric cells of cats infected with Non H. pylori Helicobacter (NHPH) and to investigate an association with the presence of inflammatory infiltrate. Forty-eight paraffin-embedded gastric samples were retrieved from the archives of the Veterinary Anatomic Pathology Laboratory that had previously been shown to be positive for NHPH with the rapid urease test and cytology. Infection by NHPH was confirmed by histopathology using the Warthin-Starry staining. Hematoxylin-eosin stained sections were reviewed to evaluate inflammatory cell infiltrates. Immunohistochemical analysis was done using anti- H. pylori antibody and anti-HER-2 antibody. Molecular analysis was performed by PCR to confirm the presence of Helicobacter. Statistical analysis was performed to determine whether there was an association between the presence of H. Heilmannii and HER-2 expression in gastric samples. All samples were positive for NHPH, by immunohistochemistry, and confirmed by PCR as H. Heilmannii. On histopathologic analysis, 56,3% of the samples had lymphocytes and plasma cells infiltrates, 52,1% of which were mild and 4,2% moderate. The intensity of the inflammatory infiltrate in the gastric mucosa was significantly greater in the complete plasma membrane of parietal cells of gastric glands that had greater HER-2 immunoexpression (p = 0.0001). A statistically significant association (p = 0.007) between the H. Heilmannii infection score and the expression of HER-2 in the lateral membrane of gastric surface cells was observed. HER-2 expression may be increased in feline gastric cells infected by H. Heilmannii and in parietal cells of gastric glands with an increased inflammatory infiltrate.

    更新日期:2019-11-01
  • Expression and tissue distribution analysis of Angiotensin II in sheep (Ovis aries) skins associated with white and black coat colors.
    Acta Histochem. (IF 1.719) Pub Date : 2019-03-20
    Zhihong Yin,Yaming Ge,Hongmei Ning,Yubo Zhu,Lingli Chen,Shouping Zhang,Xiaojing Xia,Xinzhuang Wang,Lirui Wang,Quanhai Pang,Xingyou Liu

    Angiotensin II (AngII) regulates pigment synthesis by tyrosinase in melanocytes. To evaluate the association between AngII and coat color formation, we detected the expression distribution of AngII in white and black sheep skins by LC-ESI-MS/MS, western blot, quantitative real-time-PCR (qPCR) and distribution of AngII by immunohistochemistry.Liquid chromatography-electrospray ionization tandem MS (LC-ESI-MS/MS) results showed that AngII was found in white and black skin tissues of sheep. Western blot results verified the LC-ESI-MS/MS results and suggested that AngII was expressed at significantly higher levels in black sheep skins compared with the white sheep skins. Quantitative real time PCR (qRT-PCR) results also revealed that the expression level of AngII mRNA was higher in black sheep skins than that in white sheep skins. Immunohistochemical analysis further demonstrated that AngII protein was localized in the hair bulb and outer root sheath of hair follicle in sheep. In summary, protein and transcripts exhibited the same expression pattern in white and black sheep skins. Furthermore, the expressions of AngII in the hair bulb and outer root sheath of black sheep were stronger than those in white sheep. These results suggested that AngII functions in sheep coat color regulation and offer a novel insight for further investigation on the role of AngII in the coat color formation in sheep.

    更新日期:2019-11-01
  • Inhibition of macrophage viability by bound and free bisphosphonates.
    Acta Histochem. (IF 1.719) Pub Date : 2019-03-11
    Somying Patntirapong,Pawinee Phupunporn,Daranee Vanichtantiphong,Watcharawee Thanetchaloempong

    INTRODUCTION Long-term administration of bisphosphonates (BPs) may cause osteonecrosis of the jaw (BRONJ). After administration, 50% of BPs in the circulation rapidly binds to calcium phosphate of bone. Two forms, bound and free BPs, may affect cells residing in bone including macrophages. Therefore, the aim of this study was to examine the effects of bound and free BPs on macrophage viability. MATERIALS AND METHODS Biomaterials coated with BPs were used as a model to investigate the effect of bound BPs. For free BPs, RAW cells were plated on uncoated materials and BPs were added into the media. Cell viability and number were investigated by MTT assay and nuclei staining, respectively. Furthermore, coating and washing media were collected and were used to examine cell viability. RESULTS RAW cells grew on biomaterials for 7 days. At 3 days, free and calcium-bound BPs significantly decreased cell viability and cell number compared to control. Coating media collected from pre-incubation with BP-coated composite materials reduced macrophage cell viability. CONCLUSION This study showed that macrophages were directly affected by bound and free BPs. The presence of macrophages is mandatory for bone healing, thus the inhibition of cell viability might serve as an etiology of BRONJ.

    更新日期:2019-11-01
  • Intussusceptive lymphangiogenesis in vascular transformation of lymph node sinuses.
    Acta Histochem. (IF 1.719) Pub Date : 2019-03-10
    Lucio Díaz-Flores,Ricardo Gutiérrez,Mª Pino García,Miriam González-Gómez,Lucio Díaz-Flores,José Luis Carrasco,Hugo Álvarez-Argüelles

    Numerous lymphatic anastomosing channels in the lymph nodes are the most demonstrative finding of the rare lesion termed "vascular transformation of lymph node sinuses" (VTS). The mechanism of lymphatic vessel formation in VTS has not been studied. Vessel intussusception contributes to vascular expansion, and intraluminal pillars/posts, interstitial tissue structures or larger pillars (ITSs) and folds are the hallmarks of this process in blood vessels. The aim of this work is to assess whether these hallmarks of intussusception occur in VTS lymphatic vessels, indicating intussusceptive lymphangiogenesis. For this purpose, specimens of five cases of VTS were used for serial histological sections, immunohistochemistry and immunofluorescence in confocal microscopy, which enabled us to demonstrate the 3D image that defines the pillars. The studies showed a) meshworks of lymphatic vessels, which form complex loops, resembling sinuses of lymph nodes, b) presence of intralymphatic pillars, ITSs and folds, with a cover of lymphatic endothelial cells expressing podoplanin and a varying-sized connective core (e.g. collagen), and c) increase of vessel meshwork and linear arrangement, splitting and fusion of ITSs, pillars and folds, with remodelling and segmentation. In conclusion, the development of lymphatic vessel loops, ITSs, pillars and folds with segmentation in VTS supports intussusceptive lymphangiogenesis. This mechanism of intussusception is of interest because it participates in VTS histogenesis, contributes to general knowledge of intussusceptive lymphangiogenesis, which has received less attention than intussusception in blood vessels, and provides a basis for further studies in other lymphatic conditions.

    更新日期:2019-11-01
  • Ultrastructure of the dorsal claustrum in cat. II. Synaptic organization.
    Acta Histochem. (IF 1.719) Pub Date : 2019-03-09
    Dimka Hinova-Palova,Boycho Landzhov,Alexandar Iliev,Georgi Kotov,Stancho Stanchev,Vidin Kirkov,Georgi P Georgiev,Lawrence Edelstein,Adrian Paloff

    The claustrum is a bilateral subcortical nucleus situated between the insular cortex and the striatum in the brain of all mammals. It consists of two embryologically distinct subdivisions - dorsal and ventral claustrum. The claustrum has high connectivity with various areas of the cortex, subcortical and allocortical structures. It has long been suggested that the various claustral connections have different types of synaptic contacts at the claustral neurons. However, to the best of our knowledge, the literature data on the ultrastructural organization of the different types of synaptic contacts in the dorsal claustrum are very few. Therefore, the aim of our study was to observe and describe the synaptic organization of the dorsal claustrum in the cat. We used a total of 10 adult male cats and conducted an ultrastructural study under a transmission electron microscope as per established protocol. We described a multitude of dendritic spines, which were subdivided into two types - with and without foot processes. Based on the size and shape of the terminal boutons, the quantity and distribution of vesicles and the characteristic features of the active synaptic zone, we described six types of synaptic boutons, most of which formed asymmetrical synaptic contacts. Furthermore, we reported the presence of axo-dendritic, axo-somatic, dendro-dendritic and axo-axonal synapses. The former two likely represent the morphological substrate of the corticoclaustral pathway, while the remaining two types have the ultrastructural features of inhibitory synapses, likely forming a local inhibitory circuit in the claustrum. In conclusion, the present study shares new information about the neuropil of the claustrum and proposes a systematic classification of the types of synaptic boutons and contacts observed in the dorsal claustrum of the cat, thus supporting its key and complex role as a structure integrating various information within the brain.

    更新日期:2019-11-01
  • 更新日期:2019-11-01
  • Histopathological effects of sildenafil citrate on rat corpus cavernosum.
    Acta Histochem. (IF 1.719) Pub Date : 2004-03-23
    Bilal Gümüş,H Seda Vatansever,Talha Müezzinoğlu,Sevda Müftüoğlu,Figen Kaymaz,Coşkun Büyüksu

    Sildenafil citrate (Viagra) is widely used for the treatment of erectile dysfunction with various etiologies. The aim of the present study was the investigation of histopathological effects of sildenafil citrate on rat corpus cavernosum using tight and electron microscopical techniques. Twenty male rats were divided into two groups. The first group (n = 10) was used as a control and the second group (n = 10) was treated with sildenafil citrate. Penile tissue was collected, fixed with formalin and embedded in paraffin for light microscopy, or fixed with gluteraldehyde and osmium tetroxide and embedded in Epon for electron microscopy. Light microscopical analysis showed that the corpus cavernosum was elongated and the number of blood vessels was increased. The amount of connective tissue in the penis was increased and dense collagen and smooth muscle fibers were observed in treated rats. Electron microscopical analysis showed that stromal structures of the corpus cavernosum (collagen fibers and number of cellular elements) were increased in treated rats. Fibroblasts showed signs of activation and the number of other stromal cells was increased. Immature newly synthesized collagen fibers were observed and penetrated endothelial basement membranes. In addition, endothelial cells also showed signs of activation such as cytoplasmic granules in treated rats, whereas the surface area of blood vessels was increased and basement membranes were thickened. These histopathological changes due to treatment with sildenafil citrate indicate that prolonged use of sildenafil citrate may increase the risk of fibrosis in the penis.

    更新日期:2019-11-01
  • The charged milieu: a major player in fertilization reactions.
    Acta Histochem. (IF 1.719) Pub Date : 2004-03-23
    Marcela Maldonado,Gayani Weerasinghe,Fabienne Ambroise,Edward Yamoah,Monica Londono,Juan Carlos Pelayo,Melina Grigorian,Steven B Oppenheimer

    In previous studies, we have found that negatively charged, but not uncharged, amino acids and sugars block sea urchin fertilization. These studies were developed from modeling work in non-living systems using derivatized agarose beads that suggested that charge-charge bonding may control at least some adhesive interactions. In the present study, the effects of positively charged, negatively charged and uncharged molecules were examined in the sea urchin sperm-egg system in over 300 individual trials. The results indicate that depending on the specific molecules utilized, both sperm and egg are exquisitely sensitive to charged but not uncharged molecules and to pH changes in sea water caused by some of the charged molecules. It is shown that egg activation, as well as sperm motility and sperm-egg interactions, can be affected by charged molecules. One compound, fructose-1-phosphate blocked fertilization in S. purpuratus sea urchins but not in Lytechinus pictus sea urchins. These findings indicate that charge alone cannot explain all the results. In this case, the presence of a ketone instead of an aldehyde group indicates that species-specific components may control fertilization reactions. The present study is a comprehensive survey of the effects of charge, pH and molecular structure on the fertilization activation continuum in a model system of sea urchins.

    更新日期:2019-11-01
  • Review article--involvement of gastric APUD cells in chronic renal failure.
    Acta Histochem. (IF 1.719) Pub Date : 2003-12-06
    Irena Kasacka

    Uremia leads to a number of metabolic and hormonal disorders induced by renal failure with definite biological and clinical sequels. Most frequently, alimentary disorders are the first to appear, followed by symptoms from other organs and systems. The gastrointestinal tract is a site of synthesis of many compounds that have hormonal or hormonal-like biological activity. These substances are produced by highly-specialised receptor-effector cells, that are dispersed in the gastrointestinal mucosa and classified as APUD cells. The present review is an attempt to make a synthesis of current opinions and views concerning the effect of homeostatic dysfunction of the kidneys on the morphology and action of APUD cells in the stomach.

    更新日期:2019-11-01
  • Distribution patterns of TGF-alpha, laminin and fibronectin and their relationship with folliculogenesis in rat ovary.
    Acta Histochem. (IF 1.719) Pub Date : 2003-12-06
    Gökhan Akkoyunlu,Ramazan Demir,Ismail Ustünel

    Many growth factors are considered to be involved in regulatory functions in the ovary. Specific factors mediate local cell-cell interactions in relation to follicle development. As a result of the complexity of the estrous cycles in experimental models, it is not easy to determine the role of a growth factor such as transforming growth factor alpha (TGF-alpha) in the system. Moreover, little is known about possible interactions of TGF-alpha and laminin and fibronectin in basement membranes during estrous cycles in relation to follicle development. Therefore, the present study was designed to investigate distribution patterns of TGF-alpha, laminin and fibronectin and their possible roles during follicle maturation in normal rat ovary. Ovaries were obtained from 6 adult virgin female rats and fixed in buffered neutral formalin. TGF-alpha, laminin, and fibronectin distribution patterns were evaluated using 5-7-microm-thick serial sections using the immunoperoxidase method. It was found that TGF-alpha was predominantly localised in nuclei of oocytes. Varying amounts of TGF-alpha were found in granulosa cells and interstitial thecal cells which form follicles. In addition, laminin and fibronectin were found predominantly in vascular walls, outer layers of granulosa cells and basement membranes of cuboidal/columnar surface epithelium of rat ovary. Therefore, we suggest that TGF-alpha is involved in follicular maturation. Moreover, because laminin was found to be present in between parenchymal follicle cell layers, we suggest that they were attached to supportive stromal cells by fibronectin. As TGF-alpha is associated with follicles and their relationship with the extracellular matrix, TGF-alpha may also induce formation of basement membranes which contains laminin and fibronectin components.

    更新日期:2019-11-01
  • Protective effect of melatonin on beta-cell damage in streptozotocin-induced diabetes in rats.
    Acta Histochem. (IF 1.719) Pub Date : 2003-09-19
    Ozlem Yavuz,Meryem Cam,Neslihan Bukan,Aysel Guven,Fatma Silan

    The aim of the present study was the evaluation of possible protective effects of melatonin against beta-cell damage in streptozotocin-induced diabetes in rats. Malondialdehyde levels and glutathione peroxidase activity were measured in pancreatic homogenates. Pancreatic beta-cells were examined by immunohistochemical methods. Streptozotocin was injected intraperitoneally at a single dose of 60 mg/kg for induction of diabetes. Melatonin (200 microg/kg/day, ip) was injected for 3 days prior to administration of streptozotocin; these injections were continued until the end of the study (4 weeks). Streptozotocin induced a significant increase in malondialdehyde levels (p < 0.01) and a significant decrease in glutathione peroxidase activity (p < 0.05) in pancreatic tissue. Degeneration of islet cells and weak immunohistochemical staining of insulin was observed in diabetic rats. Treatment of diabetic rats with melatonin markedly reduced malondialdehyde production (p < 0.05) and increased glutathione peroxidase activity (p < 0.01) without affecting hyperglycemia. Increased staining of insulin and preservation of islet cells were apparent in the melatonin-treated diabetic rats. These data suggest that melatonin treatment has a therapeutic effect in diabetes by reduction of oxidative stress and preservation of pancreatic beta-cell integrity.

    更新日期:2019-11-01
  • Immunolocalization of alphaV, alpha3 and beta1 integrins in the human placenta with pre-eclampsia.
    Acta Histochem. (IF 1.719) Pub Date : 2003-09-19
    H Seda Vatansever,V Sevinc Inan,Selman Lacin,Faik Koyuncu

    Signs of pre-eclampsia are considered to be caused by maternal endothelial dysfunction due to circulating factors of placental origin. Integrins are a large family of cell surface, proteins that serve as receptors involved in cell-cell and cell-matrix interactions during placentation. Therefore, low expression of integrins or the lack of it may be encountered during pre-eclampsia. In the present study, we investigated the immunolocalisation of integrins alphaV, alpha3 and beta1 in placentas of normal and pre-eclamptic women. Thirty-two placentas from pre-eclamptic (n = 14) and normotensive (n = 18) women were used. Immunohistochemical staining was performed on formalin-fixed, paraffin-embedded tissue specimens, using anti-alphaV, anti-alpha3 and anti-beta1 antibodies and the indirect immunoperoxidase technique. A semi-quantitative grading system (HSCORE) was used to compare immunohistochemical staining intensities. Distribution patterns of alphaV, alpha3 and beta1 integrins were detected in cytotrophoblasts and Hofbauer cells in normal and pre-eclamptic placentas. Immunostaining of alphaV and beta1 integrins was slightly decreased in pre-eclamptic samples but alpha3 integrin immunostaining was similar in pre-eclamptic and normal placentas. Decreased immunostaining of integrins in the cytotrophoblasts may considered to be a structural basis for decreased placental perfusion in pre-eclampsia.

    更新日期:2019-11-01
  • Beta1-integrin and IL-1alpha expression as bystander effect of medium from irradiated cells: the pilot study.
    Acta Histochem. (IF 1.719) Pub Date : 2003-09-19
    Jan Osterreicher,Jirí Skopek,Juta Jahns,Guido Hildebrandt,Jan Psutka,Zdenka Vilasová,Judith Maria Tanner,Jürgen Vogt,Tilman Butz

    Bystander effects have been proposed as a third action pathway of ionising radiation besides direct and indirect effects. The purpose of the study was to investigate whether expression of interleukin-1alpha (IL-1alpha) and beta1-integrin is elevated in bystander cells as a marker for bystander effects in comparison with classical markers such as the clonogenic assay, apoptosis and the presence of micronuclei. The hybrid cell line E.A. hy.926 obtained by fusion of HUVEC cells with the epithelial cell line A 459 was irradiated with 0-5 Gy. Bystander effects were established via medium transfer at 45 min and 4 h after irradiation from irradiated to nonirradiated cell populations. In order to exclude effects of the irradiated medium itself, irradiated medium only was also used for transfer to nonirradiated cells. Then, cells were fixed at 1, 2, 6, and 24 h after irradiation or medium transport and IL-1alpha and beta1-integrin were detected and evaluated. A higher number of beta1-integrin-positive cells was observed in both irradiated and bystander cell populations than in the control group at 1 and 24 h after irradiation with 1 Gy or medium transfer. Significantly higher numbers of IL-1alpha-positive cells were found at 1, 2, and 6 h after irradiation with 1 Gy or medium transfer as well as at 2 and 6 h after irradiation with 5 Gy or medium transfer. Clonogenic survival decreased dependently on the dose in irradiated cells but did not show any significant difference between the bystander cell populations and sham-irradiated cells. The irradiated medium itself did not have any effect. It is concluded that beta1-integrin and IL-1alpha expression may serve as more sensitive markers of post-irradiation responses in bystander cell populations than the classical radiobiological markers. Moreover, overexpression of beta1-integrin and IL-1alpha may induce increased susceptibility to inflammation of bystander cells.

    更新日期:2019-11-01
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