Mucinous adenocarcinoma as heterologous element in intermediately differentiated Sertoli–Leydig cell tumor of the ovary

Sertoli–Leydig cell tumor (SLCT) is a rare tumor involving the ovary. Approximately 20% of SLCT are associated with heterologous elements that are either of endodermal or mesodermal origin. The gastrointestinal-type epithelium is the most commonly described endodermal heterologous element. SLCT with benign and borderline mucinous neoplasm has been reported in the literature. However, SLCT with mucinous adenocarcinoma as heterologous element has been rarely documented. Herein, we describe a rare case of intermediately differentiated Sertoli–Leydig cell tumor with mucinous adenocarcinoma as the heterologous element in a 21-year-old woman. She presented with throbbing lower abdominal pain and was found to have a large, complex left ovarian mass on imaging studies. She underwent left salpingo-oophorectomy, appendectomy and lymph node staging. Gross examination of the surgical specimen showed a large, encapsulated, solid-cystic mass completely replacing the ovary. Microscopically, the tumor was composed of intermediately differentiated Sertoli–Leydig cell tumor and well-differentiated mucinous adenocarcinoma. Interestingly, the bulk of the tumor (more than 90%) was composed of mucinous adenocarcinoma, whereas the SLCT component comprised less than 10% of the total tumor. The mucinous adenocarcinoma expressed positivity for CK20, CEA, CDX2 and CK7, and the SLCT component was positive for inhibin expression. The histopathological features and results of immunostaining were consistent with the diagnosis of the intermediately differentiated SLCT with mucinous adenocarcinoma as the heterologous element. This case was a diagnostic challenge as more than 90% of the tumor was composed of mucinous adenocarcinoma and SLCT constituted only the minor part of the tumor. This feature was in contrast to the previously described two cases, where mucinous adenocarcinoma as heterologous element was present as microscopic foci. This case highlights the importance of identifying the SLCT component in a case of an apparently pure mucinous adenocarcinoma in a young patient.     

The numbers of leukocyte subsets in lung sections differ between intercellular adhesion molecule-1–/–, lymphocyte function-associated antigen-1–/– mice and intercellular adhesion molecule-1–/– mice after aerosol exposure to Haemophilus influenzae type-b

In order to investigate the role of the adhesion molecules intercellular adhesion molecule-1 (ICAM-1) and lymphocyte function-associated antigen-1 (LFA-1) in pulmonary immunological processes, leukocyte populations were stained immunohistochemically on cryostat lung sections of ICAM-1-/- and LFA-1-/- mice. A further group of ICAM-1-/- mice was exposed to Haemophilus influenzae type-b (Hib) 24 h before being sacrificed. Comparison of the numbers of leukocytes in these groups revealed different behaviors of the leukocyte subsets: granulocytes were significantly increased in all three groups. Lymphocytes were increased in ICAM-1-/- mice, while there was no significant difference in LFA-1-/- and even a decrease in ICAM-1-/- mice after Hib exposure. Neither in ICAM-1-/- nor in LFA-1-/- mice did macrophages and dendritic cells (DCs) show significant differences to control animals. After Hib exposure, a significant elevation of DCs was observed. The following conclusions can be drawn: (1) all investigated leukocyte subsets can use ICAM-1- and LFA-1-independent pathways in the lungs of mice; (2) the pathways used by the leukocytes are cell-type specific; (3) ICAM-1 plays an important role in the enhanced recruitment of lymphocytes during Hib challenge in the lung; and (4) the alternative migratory mechanisms are able to compensate for the absence of ICAM-1 or LFA-1 or even lead to increased cell numbers. This overcompensation can be seen as a result of a balance between active alternative migratory mechanisms, which takes place in the absence of ICAM-1 or LFA-1.     

High-throughput analysis of T cell–monocyte interaction in human tuberculosis

The lack of efficient tools for identifying immunological correlates of tuberculosis (TB) protection or risk of disease progression impedes the development of improved control strategies. To more clearly understand the host response in TB, we recently established an imaging flow cytometer-based in-vitro assay, which assesses multiple aspects of T cell–monocyte interaction. Here, we extended our previous work and characterized communication between T cells and monocytes using clinical samples from individuals with different TB infection status and healthy controls from a TB endemic setting. To identify T cell–monocyte conjugates, peripheral blood mononuclear cells (PBMC) were stimulated with ds-Red-expressing Mycobacterium bovis bacille Calmette–Guérin or 6-kDa early secreted antigenic target (ESAT 6) peptides for 6 h, and analyzed by imaging flow cytometer (IFC). We then enumerated T cell–monocyte conjugates using polarization of T cell receptor (TCR) and F-actin as markers for synapse formation, and nuclear factor kappa B (NF-κB) nuclear translocation in the T cells. We observed a reduced frequency of T cell–monocyte conjugates in cells from patients with active pulmonary tuberculosis (pTB) compared to latent TB-infected (LTBI) and healthy controls. When we monitored NF-κB nuclear translocation in T cells interacting with monocytes, the proportion of responding cells was significantly higher in active pTB compared with LTBI and controls. Overall, these data underscore the need to consider multiple immunological parameters against TB, where IFC could be a valuable tool.     

Monosodium urate crystals induced ICAM-1 expression and cell–cell adhesion in renal mesangial cells: Implications for the pathogenesis of gouty nephropathy

BackgroundRenal disease is prevalent in gouty patients and monosodium urate (MSU) crystal deposition in the kidney can be detected in some gouty nephropathy patients. MSU crystals can induce inflammatory events, we investigated the MSU-induced expression of intercellular adhesion molecule (ICAM)-1 on human renal mesangial cells (HRMCs) and the involved signal transduction mechanisms.MethodsThe HRMCs cell line was purchased from ScienCell Research Laboratories. MSU crystals were made by dissolving uric acid in sodium hydroxide (NaOH) solution. The involvement of MAPKs, apoptosis-associated speck-like protein containing a CARD domain (ASC), and Toll-like receptor (TLR) was investigated using pharmacological inhibitors, transfection with short hairpin RNA (shRNA), or monoclonal antibodies. Protein expression was evaluated by Western blotting. The functional activity of ICAM-1 was evaluated with cell–cell adhesion assay and immunofluorescence analysis.ResultsMSU stimulation increased expression of ICAM-1 and adhesion between HRMCs and human monocytic THP-1 cells. The interaction between HRMCs and THP-1 was suppressed by ICAM-1 neutralizing antibodies. MSU stimulation induced activation of mitogen-activated protein kinases, including c-Jun N-terminal kinase (JNK), p38, and extracellular signal-regulated kinase (ERK), but only p38 was responsible for MSU-induced expression of ICAM-1 and cell–cell adhesion. ASC also play a role in MSU-induced effects. Pretreatment with monoclonal antibodies against toll-like receptor (TLR)2 or TLR4 reduced MSU-induced ICAM-1 expression, cell–cell adhesion, p38 phosphorylation but the reduction of ASC activation is insignificant.ConclusionThe MSU induced ICAM-1 expression on HRMCs and cell–cell adhesion involved TLR2/4-p38-ICAM1 pathway and TLR2/4 independent ASC-p38-ICAM1 axis. These findings might partly explain the mechanisms underlying gouty nephropathy.     

Role of the hypothalamic–pituitary–thyroid axis in metabolic regulation by JNK1

The cJun N-terminal kinase 1 (JNK1) is implicated in diet-induced obesity. Indeed, germline ablation of the murine Jnk1 gene prevents diet-induced obesity. Here we demonstrate that selective deficiency of JNK1 in the murine nervous system is sufficient to suppress diet-induced obesity. The failure to increase body mass is mediated, in part, by increased energy expenditure that is associated with activation of the hypothalamic–pituitary–thyroid axis. Disruption of thyroid hormone function prevents the effects of nervous system JNK1 deficiency on body mass. These data demonstrate that the hypothalamic–pituitary–thyroid axis represents an important target of metabolic signaling by JNK1.     

Role of complement in host–microbe homeostasis of the periodontium

Complement plays a key role in immunity and inflammation through direct effects on immune cells or via crosstalk and regulation of other host signaling pathways. Deregulation of these finely balanced complement activities can link infection to inflammatory tissue damage. Periodontitis is a polymicrobial community-induced chronic inflammatory disease that can destroy the tooth-supporting tissues. In this review, we summarize and discuss evidence that complement is involved in the dysbiotic transformation of the periodontal microbiota and in the inflammatory process that leads to the destruction of periodontal bone. Recent insights into the mechanisms of complement involvement in periodontitis have additionally provided likely targets for therapeutic intervention against this oral disease.     

The regulatory mechanism of β-eudesmol is through the suppression of caspase-1 activation in mast cell–mediated inflammatory response

β-Eudesmol is sesquiterpenoid alcohol which contains the rhizome of Atractylodes lancea. Although it has multiple pharmacological effects, the anti-inflammatory effect of β-eudesmol and its molecular mechanisms are poorly elucidated. In this study, we investigated the regulatory mechanism of β-eudesmol on mast cell–mediated inflammatory response. The results indicated that β-eudesmol inhibited the production and expression of interleukin (IL)-6 on phorbol 12-myristate 13-acetate and calcium ionophore A23187-stimulated human mast cell (HMC). In activated HMC-1 cells, β-eudesmol suppressed activation of p38 mitogen-activated protein kinase (MAPKs) and nuclear factor-κB. In addition, β-eudesmol suppressed the activation of caspase-1 and expression of receptor-interacting protein-2. These results provide new insights into the pharmacological actions of β-eudesmol as a potential molecule for use in therapy in mast cell–mediated inflammatory diseases.     

Activation of epithelial and myoepithelial cells in the salivary glands of patients with Sjögren's syndrome: high expression of intercellular adhesion molecule-1 (ICAM.1) in biopsy specimens and cultured cells

ICAM.1 (CD54) is a surface protein expressed on epithelial and other nonhematopoietic cells upon activation and is known to play an important role in the stimulation of T cells by the provision of cellular adhesion and costimulatory support. Sjogren's syndrome (SS) is an autoimmune exocrinopathy, which is characterized by chronic lymphocytic infiltration of exocrine glands and aberrant activation of epithelial tissues. To address the contribution of ICAM.1 in the pathogenesis of SS, the expression of this protein was studied by immunohistochemistry and flow cytometry in minor salivary gland (SG) biopsies as well as in cultured SG epithelial cell (SGEC) lines obtained from 18 SS patients and 16 controls. In biopsies from SS patients (but not controls), strong ICAM.1 was expressed by infiltrating mononuclear cells (52%) and by a significant proportion of periacinar myoepithelial cells (18%). In addition, a patchy pattern of moderate ICAM.1 expression was detected in 31% of ductal epithelia of SS patients. These ICAM.1-expressing epithelial and myoepithelial cells were observed throughout glandular tissues and were not confined in areas proximal to lymphoid infiltrates. In support to an intrinsic activation profile of SGEC in SS, long-term cultured non-neoplastic SGEC lines derived from SS patients displayed significantly upregulated spontaneous expression of ICAM.1, compared to controls (P < 0.05). The high expression of ICAM.1 protein by the salivary epithelium of SS patients is likely suggestive of its important role in the pathogenesis of the disorder. Further, our results support a model of intrinsic activation of salivary epithelial and myoepithelial cells in SS, whereby these cells actively participate in the induction and maintenance of lymphocytic infiltrates of patients.     

Sertoli–Leydig cell tumours of the ovary and testis: a CGH and FISH study

We present two malignant cases of Sertoli–Leydig cell tumours (SLCT) of the testis and one ovarian SLCT with benign behaviour. The DNA copy number changes affected chromosome 1, 8, 9p, 10, 11, 12, 16, 19, 22 and X. The present study is the first molecular–cytogenetic analysis of Sertoli–Leydig cell tumours of the testis.     

Ameliorative effect of α-tocopherol on polychlorinated biphenyl (PCBs) induced testicular Sertoli cell dysfunction in F1 prepuberal rats

The study was conducted to investigate the protective role of α-tocopherol against polychlorinated biphenyls (PCBs) induced effect in Sertoli cell function of F₁ prepuberal rats. Dams were grouped into six; each group consists of six animals. Group 1–control treated with corn oil as vehicle, group II- 0.5 mgPCBs/kg bw/day, group III- 0.5mgPCBs/kg bw/day with α-tocopherol (50 mg α-tocopherol/kg bw/day), group IV- 1mgPCBs/kg bwt/day, group V- 1mgPCBs/kg bw/day with α-tocopherol (50 mg α-tocopherol/kg bw/day) and group VI ? α-tocopherol alone treated orally from postpartum day1-20. Male offspring rats were euthanized on post natal day 21. Testes were collected for the histological examination and Sertoli cell isolation. The protein levels of follicle-stimulating hormone receptor, androgen binding protein, androgen receptor, estrogen receptor α & β, Inhibin-β, transferrin, claudin-11, occludin, E-cadherin, connexin-43, c-fos, c-jun, SF1, USF1 & 2 were studied using western blot method. The testicular architecture was affected in the PCBs exposed rats but this effect was restored by α-tocopherol supplementation. PCBs decreased the protein levels of FSHR, AR, ABP, ERα & β, transferrin, claudin-11, occludin, E-cadherin, connexin-43, c-fos, c-jun, SF1, USF1 & 2 whereas inhibin-β protein level was found to be increased in Sertoli cells. These results suggested that α-tocopherol has ameliorative role against PCBs induced testicular Sertoli cell dysfunction in F₁ progeny.     

Does the Arg-Gly-Asp (RGD) adhesion sequence play a role in mediating sperm interaction with the human endosalpinx?

BACKGROUND: Sperm from several species, including the human, make direct contact with the endosalpinx. Although this is known to be beneficial to sperm function, the specific mechanisms mediating the adhesion are poorly understood. METHODS: Short linear oligopeptides containing the amino acid sequence Arg-Gly-Asp (RGD) or a scrambled sequence (GRGES) were incorporated into an established sperm-endosalpingeal binding assay. In addition, the ability of fluorescent latex beads coated with an RGD oligopeptide to bind specifically to sperm and/or epithelium was also determined. RESULTS: Significantly fewer sperm associated per field of isthmic epithelium in the presence of 62.5 micro mol/l GRGDTP (1.18 +/- 0.41; mean +/- SEM, P < 0.05) and 250 micro mol/l RGDV (1.17 +/- 0.29; P < 0.01) compared with the control incubation (3.34 +/- 0.45). There was no difference in sperm binding to ampullary epithelium in the presence of any of the oligopeptides tested. Moreover, no beads were observed bound to sperm whereas significantly more RGD-coupled beads bound to isthmic epithelium compared with ampullary epithelium (1.47 +/- 0.26 versus 0.72 +/- 0.16 P < 0.01) and this increased in a dose-dependent manner. CONCLUSIONS: These findings indicate that the recognition between the RGD sequence and integrin receptors may contribute to the interaction between sperm and the human endosalpinx in the isthmic but not in the ampullary region of the uterine tube.     

Carcinoembryonic antigen-stimulated THP-1 macrophages activate endothelial cells and increase cell–cell adhesion of colorectal cancer cells

The liver is the most common site for metastasis by colorectal cancer, and numerous studies have shown a relationship between serum carcinoembryonic antigen (CEA) levels and metastasis to this site. CEA activates hepatic macrophages or Kupffer cells via binding to the CEA receptor (CEA-R), which results in the production of cytokines and the up-regulation of endothelial adhesion molecules, both of which are implicated in hepatic metastasis. Since tissue macrophages implicated in the metastatic process can often be difficult to isolate, the aim of this study was to develop an in vitro model system to study the complex mechanisms of CEA-induced macrophage activation and metastasis. Undifferentiated, human monocytic THP-1 (U-THP) cells were differentiated (D-THP) to macrophages by exposure to 200 ng/ml phorbol myristate acetate (PMA) for 18 h. Immunohistochemistry showed two CEA-R isoforms present in both U- and D-THP cells. The receptors were localized primarily to the nucleus in U-THP cells, while a significant cell-surface presence was observed following PMA-differentiation. Incubation of D-THP-1 cells with CEA resulted in a significant increase in tumor necrosis factor-alpha (TNF-α) release over 24 h compared to untreated D-THP-1 or U-THP controls confirming the functionality of these cell surface receptors. U-THP cells were unresponsive to CEA. Attachment of HT-29 cells to human umbilical vein endothelial cells significantly increased at 1 h after incubation with both recombinant TNF-α and conditioned media from CEA stimulated D-THP cells by six and eightfold, respectively. This study establishes an in vitro system utilizing a human macrophage cell line expressing functional CEA-Rs to study activation and signaling mechanisms of CEA that facilitate tumor cell attachment to activated endothelial cells. Utilization of this in vitro system may lead to a more complete understanding of the expression and function of CEA-R and facilitate the design of anti-CEA-R therapeutic modalities that may significantly diminish the metastatic potential of CEA overexpressing colorectal tumors.     

Distribution of somatostatin-28 (1–12) immunoreactivity in the diencephalon and the brainstem of the dog

The term somatostatin refers to a family of peptides, mainly somatostatin-14, somatostatin-28 and somatostatin-28 (1–12), which are the cleavage products of a single 116 amino acid-long preprosomatostain molecule. The production of antibodies to these peptides allows their localization in a number of neuronal populations throughout the entire neuroaxis in many mammals. The dog has been pointed out as an extremely useful animal model for studying age-related cognitive dysfunction and other neuronal changes associated with aging in which somatostatin appears to be involved. However, only very scanty information is available with regard to the distribution of somatostatin in the brain of the dog. In the present work we have determined the pattern of the distribution of somatostatin-28 (1–12) immunoreactivity in the diencephalon and the brainstem of the dog. High to moderate densities of labeled perikarya were found in the anterior periventricular and arcuate hypothalamic nuclei, the reticular thalamic nucleus, in delimited parts of the nucleus of the brachium inferior colliculus, the retrorubral area, the dorsal raphe nucleus, the myelencephalic reticular formation and the dorsal motor nucleus of the vagus. Less dense population of somatostatin cells were localized in other diencephalic and brainstem nuclei. The distribution of labeled fibers was even broader as in addition to those above mentioned there were a number of areas that appeared devoid of labeled perikarya. Many of the findings were similar to those reported in earlier works while others underlined the existence of inconsistencies in the distribution pattern of this peptide in the brain of mammals.     

Serum autoantibodies profile and increased levels of circulating intercellular adhesion molecule-1: a reflection of the immunologically mediated systemic vasculopathy in rheumatic diseases?

The clinical manifestation of systemic vasculitis may be postulated as a consequence of immune response abnormalities in the course of connective tissue diseases (CTD). The aim of this study was to elucidate the significance of the different autoantibodies and soluble intercellular adhesion molecule 1 (sICAM-1) being shed into the circulation in the diagnosis of vasculitis in rheumatic diseases. Sera of 86 patients with rheumatic diseases (54 with rheumatoid arthritis (RA) and 32 with CTD) were analyzed for the concentrations of sICAM-1 levels by the enzyme-linked immunosorbent assay (ELISA). Control sera were obtained from 30 healthy individuals. Anti-nuclear antibodies (ANA), anti-double-stranded DNA (anti-dsDNA) antibodies and anti-proteinase 3 (PR-3) antibodies (cytoplasmic specific anti-neutrophil cytoplasmic autoantibodies, cANCA) were assessed by the ELISA method. Fifty out of the 86 patients had systemic lesions. A pathological picture of the vascular loop under nailfold capillary microscopy was found in 84 patients. In 19 patients the microvascular changes were advanced, in 35 moderate and in 30 mild. All patients with articular manifestations had pathological changes under capillary microscopy. Patients with advanced changes under capillary microscopy had longer disease durations than patients with a mild intensity of vasculitis. The serum concentrations of sICAM-1 were significantly increased in RA and CTD patients compared with 30 controls (in both cases p<0.001). Moreover, RA and CTD patients with systemic vasculitis showed significantly higher levels of sICAM-1 than those without vascular involvement (p<0.001 and p<0.005 respectively). ANA were observed in significantly elevated concentration among RA and CTD patients with the systemic damage compared with patients without organ injury (p<0.001 and p<0.05 respectively). Also, cANCA levels were two-fold higher, but only among CTD patients with systemic damage (p<0.05). Serum concentrations of sICAM-1 were elevated in the patients showing the presence of ANA antibodies (p<0.05). Significant correlations between ANA level and disease duration and hemoglobin concentration were observed. The concentrations of cANCA correlated with those of rheumatoid factor and of dsDNA with patient age. We conclude that systemic lesions in the course of RA and CTD are accompanied by the microvascular injury observed under nailfold capillary microscopy. Our data suggest that sICAM-1, ANA and cANCA serum levels may reflect the extent of the vascular involvement in RA and CTD patients.     

The role of β1 1 integrins in adhesion of two breast carcinoma cell lines to a model endothelium

Interactions between tumour cells and the endothelium are vital to the formation of haematogenous metastases. Binding to model endothelium of one oestrogen receptor positive breast carcinoma cell line (MCF-7) and one receptor negative line (HS578T) was examined in vitro together with endothelial retraction induced by these tumour cells. Adhesion was inhibited by monoclonal antibodies specific for the VLA integrins and by peptides containing the RGD motif which is commonly recognised as a ligand by the VLA adhesion molecules. However, binding of the two tumour cell lines was inhibited by monoclonal antibodies specific for different VLA molecules; anti-61 inhibited MCF-7 adhesion but anti-51 inhibited Hs578T. These results were consistent with flow cytometric quanfication of the expression of these VLA integrins on the surfaces of the two tumour cell lines. Enzyme-linked immunosorbent assays (ELISA) demonstrated that laminin was present on the endothelial cell surface but collagen IV was absent. ELISA failed to detect increased exposure of the subendothelial matrix during the first hour after addition of either cancer cell type. This was supported by assays which demonstrated maintenance of the endothelial permeability barrier during this period. Slight endothelial retraction was detected within 2 hours of the addition of tumour cells. It is concluded that binding between tumour cells and confluent endothelium is inhibited by the blockade of adhesion molecules which are normally associated with interactions between the cell and the subendothelial matrix. Tumour cell to matrix interactions rather than direct tumour to endothelial cell adhesion may be the limiting step in tumour cell binding to the endothelium.     

Absence of the BRAF and the GRIM-19 mutations in oncocytic (Hürthle cell) solid cell nests of the thyroid

We report the first case of oncocytic solid cell nests (SCNs), found in the right lobe of the thyroid of a 70-year-old man. Conventional SCNs and 1 papillary microcarcinoma (mPTC) were also found in the left lobe. In the oncocytic SCNs, 80% of the main cells showed oncocytic cytoplasm immunoreactive for porin and proteins of the SDHB and SDHA genes. Positivity for cytokeratin 19, p63, galectin-3, and HBME-1 and negativity for thyroglobulin, thyroperoxidase, vimentin, Oct-4, and α-fetoprotein were found in oncocytic and conventional SCNs. An inverse correlation was found between oncocytic metaplasia and p63. Association with C cells was confirmed at protein and messenger RNA levels in both types of SCNs. No germinal mutation of GRIM-19 was detected. No somatic BRAF mutation was found in any of the SCNs nor in the mPTC. We conclude that SCNs may acquire mitochondrial alterations similar to those seen in follicular and C cells, as well as in their respective tumors.