Columnar Epithelial Cells Research Articles

Immunotoxicity of β-Diketone Antibiotic Mixtures to Zebrafish (Danio rerio) by Transcriptome Analysis.

Fluoroquinolones and tetracyclines are known as β-diketone antibiotics (DKAs) because of bearing a diketone group in their molecular structure. DKAs are the most widely used antibiotics to prevent generation of disease in humans and animals and to suppress bacterial growth in aquaculture. In recent years, overuse of DKAs has caused serious environmental risk due to their pseudo-persistence in the environment, even though their half-lives are not long. So far, no reports were concerned with the joint immunotoxicity of DKAs. Herein, we reported on the immunotoxicity of DKAs on zebrafish after a 3-month DKAs exposure using transcriptomic techniques. According to transcriptome sequencing, 10 differentially expressed genes were screened out among the genes related to KEGG pathways with high enrichment. The identified 7 genes showed to be consistent between RNA-seq and qRT-PCR. Due to DKAs exposure, the content or activity for a series of immune-related biomarkers (Complement 3, lysozyme, IgM and AKP) showed the inconsistent changing trends as compared with the control group. Histopathological observations showed that the number of goblet cells increased sharply, the columnar epithelial cells swelled, the nucleus became slender in intestinal villi, and numerous brown metachromatic granules occurred in spleens of DKAs-exposed groups. Overall, both detection of biomarkers and histopathological observation corroborated that chronic DKAs exposure could result in abnormal expression of immune genes and enzymes, and variable levels of damage to immune-related organs. These complex effects of DKAs may lead to zebrafish dysfunction and occurrence of diseases related to the immune system.

Cardio‐respiratory Development in Juvenile Sprague‐Dawley Rats: a Histomorphologic and Immunohistochemical Study

Juvenile toxicity studies have become an important and crucial focus of the preclinical drug industry. To accurately distinguish the potential detrimental effects of new drugs on the cardio‐respiratory system, it is essential to thoroughly define the normal tissue remodeling features characterizing the development and maturation of this system. The purpose of this study was to investigate the key histomorphological postnatal changes of the cardio‐respiratory system in Sprague‐Dawley rats from birth to postnatal day (PnD) 30, with a special focus on cell proliferation and apoptosis. Formalin‐fixed and paraffin‐embedded lung, trachea, heart and aorta were obtained from 51 Sprague Dawley rats divided in 13 timepoints according to age (PnD1, 2, 4, 6, 8, 10, 14, 17, 21, 24, 26, 28, and 30). Whenever possible, equal numbers of females and males were used in each group. Sections were cut at 4mm thickness and stained with hematoxylin and eosin. In addition, immunohistochemical demonstration of vimentin within lung samples and distinction between collagen and elastin fibers within the aortic wall with Masson's trichrome stain was performed. When relevant, immunohistochemistry staining was performed to highlight apoptosis (caspase‐3) and cell proliferation (Ki‐67). The ratio of heart weight relative to body weight was calculated for each timepoint and was higher at birth, then progressively decreased until PnD30. The rat heart was morphologically immature at birth and developed by cell hyperplasia over the perinatal period followed by cell hypertrophy around the third postnatal week. The newborn myocardium was hypercellular with less prominent cross‐striations, and mitotic figures and apoptotic bodies were frequently seen. The aortic wall progressively increased in thickness throughout the duration of the study as a result of an accumulation of extracellular matrix (collagen and elastic fibers). At birth, the trachea was lined by immature columnar ciliated epithelial cells, and submucosal glands became visible around one week after birth. The newborn rat had no alveoli but instead possessed large gas exchange units termed “primary saccules” characterized by highly cellular walls with a central sheet of connective tissue flanked on both sides by flattened cells (primary septa). An intense interstitial cellular proliferation was observed in the lungs between PND6 and PND14 resulting in new secondary septa, which were highly vimentin immunopositive. By the end of the third postnatal week, the septa underwent noticeable thinning and the lung parenchyma progressively matured, resulting in increased gas exchange surfaces. The findings described herein suggest that the cardio‐respiratory system of rats is immature at birth. The lungs and airways reached maturity 3 weeks after birth while cardiomyocyte hypertrophy was noticed until PnD30. The data generated will serve as background historical database that will be valuable when performing postnatal developmental toxicity studies.Support or Funding InformationMitacs

Expression, localization and possible functions of aquaporins 3 and 8 in rat digestive system

Although aquaporins (AQPs) play important roles in transcellular water movement, their precise quantification and localization remains controversial. We investigated expression levels and localizations of AQP3 and AQP8 and their possible functions in the rat digestive system using real-time polymerase chain reactions, western blot analysis and immunohistochemistry. We investigated the expression levels and localizations of AQP3 and AQP8 in esophagus, forestomach, glandular stomach, duodenum, jejunum, ileum, proximal and distal colon, and liver. AQP3 was expressed in the basolateral membranes of stratified epithelia (esophagus and forestomach) and simple columnar epithelia (glandular stomach, ileum, and proximal and distal colon). Expression was particularly abundant in the esophagus, and proximal and distal colon. AQP8 was found in the subapical compartment of columnar epithelial cells of the jejunum, ileum, proximal colon and liver; the most intense staining occurred in the jejunum. Our results suggest that AQP3 and AQP8 play significant roles in intestinal function and/or fluid homeostasis and may be an important subject for future investigation of disorders that involve disruption of intestinal fluid homeostasis, such as inflammatory bowel disease and irritable bowel syndrome.

Immature teratoma of vulva in a 15-year-old girl

Backgrounds: Immature teratoma (IT) of vulva (V) has been reported only once. Case report: A 15-year-old girl presented a vulvar tumor, and tumorectomy was performed. The tumor measured 4 cm × 4 cm × 5 cm and multiply cystic. The cysts contained mucinous fluid. The tumor was well demarcated from surrounding tissues and tumorectomy was easy. Microscopically, the tumor was multicystic neoplasm composed of mucin-bearing columnar mature epithelial cells, cellular immature mesenchymal zone beneath the columnar epithelial cells, immature cartilage, and mature connective tissue containing vasculatures. Immunohistochemically, the immature mesenchymal elements of the tumor were positive for p53, Ki67 (labeling = 30%) and vimentin, but negative for smooth muscle actin, myoglobin, HHF-35 caldesmon, and S100 element. No immature neural elements were seen. Since three germ elements were seen and some showed immature features, a diagnosis of IT was made. Conclusions: A very rare case of IT of V is reported. This is the second case of IT in this location.

Histoenzymological Observations on Acid Phosphatase Activity in the Posterior Intestine of HGCL2–Treated Fish, Channa striatus

Present studies incorporate enterotoxic effects of HgCl2 on the relative distribution of acid phosphatase (ACP) activity in the posterior intestine of fish, Channa striatus. It was observed that there appears to be slightly increased ACP activity accompanied by histolytic changes in the columnar epithelial cells lining of villi and intestinal glands of the posterior intestine of treated carnivorous fish. However, in control, these changes were invisible.

Use of a feline respiratory epithelial cell culture system grown at the air–liquid interface to characterize the innate immune response following feline herpesvirus 1 infection

Infection with feline herpesvirus-1 (FHV-1) accounts for 50% of viral upper respiratory diseases in domestic cats and is a significant cause of ocular diseases. Despite the clinical significance and high prevalence of FHV-1 infection, currently available vaccines cannot completely protect cats from infection and lifelong latency. FHV-1 infects via the mucous membranes and replicates in respiratory epithelial cells, but very little is known about the early innate immunity at this site. To address questions about immunity to FHV-1, feline respiratory epithelial cells cultured at air–liquid interface (ALI-FRECs) were established by collecting respiratory tracts from 6 healthy cats after euthanasia. Cells were isolated, cultured and characterized histologically and immunologically before infection with FHV-1. The expression of Toll-like receptors (TLRs), cytokine and chemokine responses were measured by real time PCR.ALI-FRECs morphologically resembled the natural airways of cats with multilayered columnar epithelial cells and cilia. Immunological properties of the natural airways were maintained in ALI-FRECs, as evidenced by the expression of TLRs, cytokines, chemokines, interferons, beta-defensins, and other regulatory genes. Furthermore, ALI-FRECs were able to support infection and replication of FHV-1, as well as modulate transcriptional regulation of various immune genes in response to infection. IL-1β and TNFα were increased in ALI-FRECs by 24hpi, whereas expression levels of IFN-α and TLR9 were not increased until 36hpi. In contrast, TLR3, GM-CSF and TGF-1β expression was down-regulated at 36hpi.The data presented show the development of a system ideal for investigating the molecular pathogenesis and immunity of FHV-1 or other respiratory pathogens.

Poloxamer 407-based intranasal thermoreversible gel of zolmitriptan-loaded nanoethosomes: formulation, optimization, evaluation and permeation studies

Zolmitriptan is the drug of choice for migraine, but low oral bioavailability (<50%) and recurrence of migraine lead to frequent dosing and increase in associated side effects. Increase in the residence time of drug at the site of drug absorption along with direct nose to brain targeting of zolmitriptan can be a solution to the existing problems. Hence, in the present investigation, thermoreversible intranasal gel of zolmitriptan-loaded nanoethosomes was formulated by using mucoadhesive polymers to increase the residence of the drug into the nasal cavity. The preparation of ethosomes was optimized by using 32 factorial design for percent drug entrapment efficiency, vesicle size, zeta potential, and polydispersity index. Optimized formulation E6 showed the vesicle size (171.67 nm) and entrapment efficiency (66%) when compared with the other formulations. Thermoreversible gels prepared by using poloxamer 407 showed the phase transition temperature at 32–33 °C which was in line with the nasal physiological temperature. The optimized ethosomes were loaded into the thermoreversible mucoadhesive gel optimized by varying concentrations of poloxamer 407, carbopol 934, HPMC K100, and evaluated for gel strength, gelation temperature, mucoadhesive strength, in vitro drug release, and ex vivo drug permeation, where G3 and G6 were found to be optimized formulations. In vitro drug release was studied by different kinetic models suggested that G3 (n = 0.582) and G6 (n = 0.648) showed Korsemeyer–Peppas (KKP) model indicating non-Fickian release profiles. A permeation coefficient of 5.92 and 5.9 µg/cm2 for G3 and G6, respectively, revealed very little difference in release rate after 24 h between both the formulations. Non-toxic nature of the gels on columnar epithelial cells was confirmed by histopathological evaluation.

Pathological (Histological and Ultrastructural) Study in Stomach and Intestine of Heteropneustes fossilis (Bloch) to Excel Mera 71, a Glyphosate-Based Herbicide

Effects of glyphosate-based commercial herbicide, Excel Mera 71 were performed to evaluate the pathological responses in stomach and intestine of Heteropneustes fossilis (Bloch) for duration of 30 days both under rice field and laboratory concentrations. Under light microscopy, stomach showed distortion in columnar epithelial cells (CEC), lamina propria (LP) and gastric glands under both conditions, but the severity of responses were more pronounced under laboratory condition. Severe fragmentation in mucosal folds (MF) and epithelial cells, and excessive mucus secretion were observed under scanning electron microscopic (SEM) study in laboratory study but alterations in stratified epithelial cells and microridges structure were not prominent under field; while deformation in mitochondria and endoplasmic reticulum and cytoplasmic vacuolations were seen under transmission electron microscopic (TEM) observation in both the conditions, but less severe in field study. Intestine showed distortion and fatty deposition in lamina propria, lifting of CEC and loss of brush border structures in laboratory study, while damage only at the tip of the mucosal villi and CEC were observed under field study through light microscopic observation. Under SEM study, degeneration in CEC and excessive mucus secretion over CEC were prominent, while under TEM study deformation and necrosis in mitochondria, severe cytoplasmic vacuolation, necrosis, cytosolic disorganization, and loss of cellular compartmentation were observed in laboratory study, but intestinal epithelium showed normal appearance under field observation. Therefore, present investigation depicted that long-term glyphosate exposure caused stronger pathological alterations under laboratory condition compared with field study and finally, displayed responses could be considered as indicators of herbicidal contamination in aquatic environment.

Gastrointestinal Pathology in Freshwater Fish, Oreochromis niloticus (Linnaeus) Under Almix Exposure

Teleostean fish, Oreochromis niloticus was exposed to almix herbicide under field (8 g/acre) and laboratory (66.67 mg/l) conditions for a period of 30 days. Histological and ultramicroanatomical responses were investigated in stomach and intestine. In stomach, light microscopy displayed damaged Columnar Epithelial Cells (CEC) and mucosal folds, thinning of the top plate and fused mucosal folds under laboratory condition but no such significant alterations were observed except mucus secretion in field study. Electron microscopic observation revealed alterations in epithelial contour, damage in microridge structures and excessive mucin mass over epithelial surface under Scanning Electron Microscopic (SEM) study in both conditions, but the damage was more severe in laboratory study. Transmission Electron Microscopic (TEM) study disclosed severe deformed nucleus and mitochondria, cellular necrosis and severe vacuolation under laboratory study, but comparatively less symptomatic changes were observed in field study. Intestine showed damage in CEC and rupture in intestinal villi in both the conditions under light microscopy, while ultrastructural study showed degenerative changes in CEC and excess mucus secretion in laboratory condition under SEM study. TEM study depicted deformed nucleus and mitochondria, severe vacuolation, and appearance of double layered cell structures in the laboratory condition but no significant alterations were observed under SEM and TEM study in field condition. The present results clearly demonstrated comparatively less pathological lesions under field study than laboratory observations and these responses displayed by the fish species could be considered as bio indicator for evaluating herbicidal toxicosis in aquatic environment.

DNA Damage in CD133-Positive Cells in Barrett's Esophagus and Esophageal Adenocarcinoma.

Barrett's esophagus (BE) caused by gastroesophageal reflux is a major risk factor of Barrett's esophageal adenocarcinoma (BEA), an inflammation-related cancer. Chronic inflammation and following tissue damage may activate progenitor cells under reactive oxygen/nitrogen species-rich environment. We previously reported the formation of oxidative/nitrative stress-mediated mutagenic DNA lesions, 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) and 8-nitroguanine, in columnar epithelial cells of BE tissues and cancer cells of BEA tissues. We investigated the mechanisms of BEA development in relation to oxidative/nitrative DNA damage and stem cell hypothesis. We examined 8-nitroguanine and 8-oxodG formation and the expression of stem cell marker (CD133) in biopsy specimens of patients with BE and BEA by immunohistochemical analysis in comparison with those of normal subjects. CD133 was detected at apical surface of columnar epithelial cells of BE and BEA tissues, and the cytoplasm and cell membrane of cancer cells in BEA tissues. DNA lesions and CD133 were colocalized in columnar epithelial cells and cancer cells. Their relative staining intensities in these tissues were significantly higher than those in normal subjects. Our results suggest that BE columnar epithelial cells with CD133 expression in apical surface undergo inflammation-mediated DNA damage, and mutated cells acquire the property of cancer stem cells with cytoplasmic CD133 expression.

Analysis of immunohistochemical characteristics of lymphoid stroma and epithelium of the salivary gland Warthin tumor

The aim of our study was to determine using immunohistochemical method profi and relationship between lymphocytes in lymphoid rich stroma of the Warhins tumors, as well as, the relationship between lymphocytes and tumors epithelial cells. Our retrospective-prospective study we conducted in the Department of Clinical Pathology KBC Zemun on 25 samples of tissue from the parotid gland with WT tumor surgically removed at the Department of ENT KBC Zemun in the period from 2000 to 2016 year. Sections of tumor tissue, fixed in 10% formalin were stained with hematoxylin-eosin standard method, and then immunohistochemical method to: antibody B lymphocytes (CD20) antibody for T lymphocytes (CD3), keratin antibody that marks the basal and columnar epithelial cells (CK7 ) and keratin antibody that marks only the basal epithelial cells (CK5 / 6). We have found that in relation to the gender distribution of all 25 patients, there were more male patients: 16 (64%) men and 9 (36%) women. Immunohistochemical analysis with keratin antibody CK7 showed that luminal columnar cells intensely positive (+++) more than 75% in 23 cases (92%), whereas in 2 cases were intensely positive (+++) in percentages ranging from 25 to 75%. In contrast to the luminal, basal cells were always showed little immunoreactivity (+) antibody to CK7 and in 20 cases in the group of <25% of the cells respectively in the cases of 5% in the group 25-75. Evaluation of the presence, intensity, type and distribution of cells with each other and in relation to the tumor epithelial component would contribute to establishing unclear histogenesis lymphoid stroma at the tumor.

Choroid plexus papilloma in one-humped camel (Camelus dromedarius) in Sudan

The present article describes choroid plexus papilloma in a 6-year-old male dromedary camel. The main clinical manifestations were stiff gait and incoordination followed by paresis then complete paralysis of hind quarters. Postmortem revelated a mass of 2 centimetres in diameter located in a dilated 4th ventricle. Hydrocephalus was evident by multiple cystic cavities, varying in size in brain stem and cerebral hemispheres. Histopathological sections of the mass displayed proliferation of densely packed arborising papillary fronds of fine fibrovascular core covered with single to multilayer and cluster or nest of cuboidal to columnar epithelial cells. The neoplastic epithelial cells showed positive immunoreactivity to cytokeratin 7. The gross, microscopic and immunohistochemistry findings were consistent with choroid plexus papilloma (C.P.P.).

Differential expression of Toll-like receptor-2, -4 and -9 in follicle-associated epithelium from epithelia of both follicle-associated intestinal villi and ordinary intestinal villi in rat Peyer's patches.

The expressions of Toll-like receptor (TLR) -2, -4 and -9 were immunohistochemically investigated in the follicle-associated epithelium (FAE), and epithelia of the follicle-associated intestinal villus (FAIV) and ordinary intestinal villus (IV) in rat Peyer’s patch regions with no bacterial colonies on the mucous membranes. TLR-2 was expressed in the striated borders of microvillous columnar epithelial cells (MV) in both FAIV and IV except in the apices. However, TLR-2 expression in the striated borders was weaker in the epithelium of the follicular side of FAIV (f-FAIV) than in epithelia of IV and the anti-follicular side of FAIV. TLR-4 and -9 were not expressed in the FAIV and IV. In the FAE, TLR-2, -4 and -9 were not expressed in the striated borders of MV, but the roofs of some typical M-cells were immunopositive for all TLRs. Especially, no TLR-positive MV were found at the FAE sites where M-cells appeared most frequently. In the follicle-associated intestinal crypt (FAIC), immunopositivity for all TLRs was observed in the striated borders of MV and the luminal substances. In conclusion, the lower levels of TLR-2 in both FAE and the epithelium of f-FAIV probably reduce recognition of indigenous bacteria. TLR-2, -4 and -9 appear not to participate directly in differentiation of MV into M-cells, because TLRs were not expressed in any MV in the upstream region of M-cells in FAE with no settlement of indigenous bacteria in the rat Peyer’s patches.

꼬막 Tegillarca granosa 심신낭복합체의 미세해부학적 특징

ABSTRACT This study was conducted to provide the microanatomical information of the heart-kidney complex of Tegillarca granosa . The heart-kidney complex was located in the pericardial cavity between the dosal visceral mass and posterior adductor muscle. The heart composed of two atrium and one ventricle. The kidney composed of a pair of left and right. The atrium and ventricle of the heart were composed of the epicardium, myocardium and endocardium. The epicardium of simple epithelial layer composed of cuboidal epithelial cells that had a strong basophilic nucleus located in the center. The myocardium composed of muscle fiber bundles. The myocardium in ventricle was denser than in the atrium. The endocardium of simple epithelial layer composed of squamous epithelial cell that had a strong basophilic nucleus was located in the center. The endocardium thickness of the atrium was 6.04 (± 2.26) μm, endocardium thickness of the atrium was 7.36 (± 3.21) μm, and appeared to be thicker in the ventricle. The kidney composed of numerous renal tubules. The renal tubule of simple epithelial layer composed of columnar epithelial cell with nucleus located in the basal zone and a number of cytoplasmic granules. The developed striated border was the inner epidermis.Key words:

Histological, surface ultrastructural, and histochemical study of the stomach of red piranha, Pygocentrus nattereri (Kner)

Abstract The structural characterization and function of the stomach in the omnivore Pygocentrus nattereri were described using light and scanning electron microscopy. The sac-like stomach was morphologically divided into the cardiac and pyloric regions. The histological structure of the stomach consisted of four layers of the mucosa, submucosa, muscularis, and serosa. The superficial epithelium of the cardiac stomach was lined with columnar epithelial cells and the glandular epithelium contained numerous gastric glands. Gastric glands were completely absent in the pyloric portion. The mucosal surface of the stomach was a meshwork of various folds, provided with oval or rounded columnar epithelial cells which were densely packed with short, stubby microvilli. The occasional presence of conspicuous gastric pits was surrounded by epithelial cells. The localization and chemical nature of acid and neutral mucins in the various cells of the stomach was studied by employing combined the Alcian Blue-Periodic Acid Schiff (AB-PAS) technique. The deposition of glycogen was detected in the gastric glands as well as in the epithelial lining of the stomach. The utmost reactions for protein and tryptophan were recorded in the gastric glands of mucosa. The cellular organization and histochemical characterization of the stomach are discussed in relation to the feeding and digestion of the fish concerned.

An Adaptable Spectrin/Ankyrin-Based Mechanism for Long-Range Organization of Plasma Membranes in Vertebrate Tissues.

Ankyrins are membrane-associated proteins that together with their spectrin partners are responsible for micron-scale organization of vertebrate plasma membranes, including those of erythrocytes, excitable membranes of neurons and heart, lateral membrane domains of columnar epithelial cells, and striated muscle. Ankyrins coordinate functionally related membrane transporters and cell adhesion proteins (15 protein families identified so far) within plasma membrane compartments through independently evolved interactions of intrinsically disordered sequences with a highly conserved peptide-binding groove formed by the ANK repeat solenoid. Ankyrins are coupled to spectrins, which are elongated organelle-sized proteins that form mechanically resilient arrays through cross-linking by specialized actin filaments. In addition to protein interactions, cellular targeting and assembly of spectrin/ankyrin domains also critically depend on palmitoylation of ankyrin-G by aspartate-histidine-histidine-cysteine 5/8 palmitoyltransferases, as well as interaction of beta-2 spectrin with phosphoinositide lipids. These lipid-dependent spectrin/ankyrin domains are not static but are locally dynamic and determine membrane identity through opposing endocytosis of bulk lipids as well as specific proteins. A partnership between spectrin, ankyrin, and cell adhesion molecules first emerged in bilaterians over 500 million years ago. Ankyrin and spectrin may have been recruited to plasma membranes from more ancient roles in organelle transport. The basic bilaterian spectrin-ankyrin toolkit markedly expanded in vertebrates through gene duplications combined with variation in unstructured intramolecular regulatory sequences as well as independent evolution of ankyrin-binding activity by ion transporters involved in action potentials and calcium homeostasis. In addition, giant vertebrate ankyrins with specialized roles in axons acquired new coding sequences by exon shuffling. We speculate that early axon initial segments and epithelial lateral membranes initially were based on spectrin-ankyrin-cell adhesion molecule assemblies and subsequently served as "incubators," where ion transporters independently acquired ankyrin-binding activity through positive selection.

The Saccharomyces boulardii CNCM I-745 strain shows protective effects against the B. anthracis LT toxin.

The probiotic yeast Saccharomyces boulardii (S. boulardii) has been prescribed for the prophylaxis and treatment of several infectious diarrheal diseases. Gastrointestinal anthrax causes fatal systemic disease. In the present study, we investigated the protective effects conferred by Saccharomyces boulardii CNCM I-745 strain on polarized T84 columnar epithelial cells intoxicated by the lethal toxin (LT) of Bacillus anthracis. Exposure of polarized T84 cells to LT affected cell monolayer integrity, modified the morphology of tight junctions and induced the formation of actin stress fibers. Overnight treatment of cells with S. boulardii before incubation with LT maintained the integrity of the monolayers, prevented morphological modification of tight junctions, restricted the effects of LT on actin remodeling and delayed LT-induced MEK-2 cleavage. Mechanistically, we demonstrated that in the presence of S. boulardii, the medium is depleted of both LF and PA sub-units of LT and the appearance of a cleaved form of PA. Our study highlights the potential of the S. boulardii CNCM I-745 strain as a prophylactic agent against the gastrointestinal form of anthrax.

Phytoestrogenic activity of blackcurrant (Ribes nigrum) anthocyanins is mediated through estrogen receptor alpha.

Blackcurrants (Ribes nigrum L., Grossulariaceae) contain high amounts of anthocyanin polyphenols, which have antioxidant and anti-carcinogenic health benefits. This study analyzed the potential phytoestrogenic effects of blackcurrant extract (BCE) in breast cancer (MCF-7) and human endometrial cancer (Ishikawa) cell lines that over-express estrogen receptor alpha (ERα), as well as in immature female rats. Microarray analysis and Ingenuity® Pathway Analysis showed that BCE activated the ERα pathway, whereas quantitative-PCR confirmed that BCE and four types of anthocyanins up-regulated genes downstream of ERα. BCE (0.1-1.0 μg/mL) and anthocyanins (0.1-10 μM) induced MCF-7 cell proliferation; however, this effect was blocked by ER antagonist fulvestrant. Flow cytometry showed that anthocyanins reduced and increased the number of MCF-7 cells in the G0/G1 and G2/M phases, respectively. Anthocyanins stimulated ERα transcriptional activity in human ERα reporter assays and induced alkaline phosphatase activity in Ishikawa cells. Competition assays and in silico analysis indicated that anthocyanins bind to ERα. Finally, BCE focally induced stratification of columnar epithelial cells in the rat uterus and increased cytoplasmic mucin levels in these cells. These results suggest that blackcurrant anthocyanins act as phytoestrogens in vitro and in vivo.

Hormone concentration, metabolic disorders and immunoexpression of androgen and estrogen-alpha receptors in men with benign prostatic hyperplasia and testosterone deficiency syndrome.

A slight decrease in blood testosterone level in men is a physiological state associated with the aging. The aim of our study was to evaluate the occurrence of hormone and metabolic disorders, as well as the immunolocalization and immunoexpression of androgen receptors (AR) and estrogen-alpha receptors (ERa) in the prostates of men with benign prostatic hyperplasia (BPH) and coexisting testosterone deficiency syndrome (TDS). The study involved 150 men, diagnosed with and receiving pharmacological treatment for BPH. Concentrations of glucose, total cholesterol (TCh), high-density lipoproteins (HDL), low-density lipoproteins (LDL), and triglycerides (TG) were determined in blood serum. Serum concentrations of total testosterone (TT), free testosterone (FT), estradiol (E2), luteinizing hormone (LH), insulin (I), sex hormone-binding globulin (SHBG), dehydroepiandrosterone sulfate (DHEAS), and insulin-like growth factor 1 (IGF-1) were measured by ELISA. The number of AR-positive cells and ERa-positive cells were measured in prostate sections of men with BPH. Patients eligible for transurethral resection of the prostate and TDS were significantly more likely to have higher abdominal circumference and higher serum levels of insulin and IGF-1 as well as lower levels of FT and SHBG than control subjects with BPH and no TDS. Quantitative analysis revealed 35.8% AR-positive colum-nar epithelial cells and 24.3% AR-positive stromal cells in prostates of BPH patients with TDS and 30.5% and 23.0%, respectively, in BPH patients without TDS. However, the differences between the study and the control groups were statistically not significant. In prostates of BPH patients with TDS the immunoexpression of ERa was observed in 2.88% of the columnar epithelial cells and 0.39% of stromal cells. In BPH patients without TDS ERa-positive cells were only found in 0.04% of columnar epithelial cells and 0.62% of prostatic stromal cells. Considering the statistically significantly higher levels of I and IGF-1 and larger abdominal circumference of men with BPH and TT deficiency, it can be supposed that visceral obesity and carbohydrate disorders may contribute to the reduction of testosterone concentration. The results of our study indicate a relationship between TT concentration in the plasma of patients with BPH and the percentage of AR-positive cells in the prostate.

Salmonella Typhimurium exploits inflammation to its own advantage in piglets.

Salmonella Typhimurium (S. Typhimurium) is responsible for foodborne zoonotic infections that, in humans, induce self-limiting gastroenteritis. The aim of this study was to evaluate whether the wild-type strain S. Typhimurium (STM14028) is able to exploit inflammation fostering an active infection. Due to the similarity between human and porcine diseases induced by S. Typhimurium, we used piglets as a model for salmonellosis and gastrointestinal research. This study showed that STM14028 is able to efficiently colonize in vitro porcine mono-macrophages and intestinal columnar epithelial (IPEC-J2) cells, and that the colonization significantly increases with LPS pre-treatment. This increase was then reversed by inhibiting the LPS stimulation through LPS antagonist, confirming an active role of LPS stimulation in STM14028-intracellular colonization. Moreover, LPS in vivo treatment increased cytokines blood level and body temperature at 4 h post infection, which is consistent with an acute inflammatory stimulus, capable to influence the colonization of STM14028 in different organs and tissues. The present study proves for the first time that in acute enteric salmonellosis, S. Typhimurium exploits inflammation for its benefit in piglets.