Histology and histopathology Vol.29, nº 7 (2014)

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    Histological and immunohistochemical study of an unusual type of gallbladder duplication
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Carriel, Víctor; Aneiros Fernández, José; Ruyffelaert, Muriel; Arias Santiago, Salvador; Riady, Vadir; Izquierdo Martínez, Francisco; Roda, Olga; Cornelissen, María; Campos, Antonio; Alaminos, Miguel
    Gallbladder duplication is a rare congenital anomaly, with an incidence of 1 in 3,800 autopsies. The correct diagnosis and treatment of this type of entity is important in clinical practice, because it may cause some clinical and surgical problems. In this report, we present the clinical case of a 28-year-old female with abdominal pain. Ultrasound of the upper abdomen showed a distended gallbladder with the presence of a septum that could suggest a congenital anomaly of the extrahepatic biliary system. During surgery, a distended and inflamed gallbladder with a lithiasis was found. In addition, a complete septum and double cystic duct were observed. The gross and histopathological evaluation of the surgical specimen allowed us to confirm the diagnosis of a Y- shaped type gallbladder duplication according to Boyden’s classification. In conclusion, in presence of an atypical imaging of the gallbladder, diagnosis of this group of congenital anomalies should be considered in order to adequately plan surgical intervention if necessary.
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    Loss of connexin43 (Cx43) in Sertoli cells leads to spatio-temporal alterations in occludin expression
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Gerber, Jonathan; Weider, Karola; Hambruch, Nina; Brehm, Ralph
    Within the testis, Sertoli cell (SC) junctional complexes between somatic SC create a basal and apical polarity within the seminiferous epithelium, restrict movement of molecules between cells, and separate the seminiferous epithelium into a basal and adluminal compartment. This barrier consists of membrane integrated proteins known as tight, adherens, and gap junctions, which promote cell-cell contact along the blood-testis-barrier (BTB). Nevertheless, these junctions, which form the basis of the BTB are structures whose function and dynamic regulation is still poorly understood. Thus, in this study, through the use of immunohistochemistry (IHC), semi quantitative western blot (WB) analysis, and real-time-quantitative-PCR (qRT-PCR) we focused on the expression pattern of the main testicular tight junction protein, occludin, in SC. For this, the established transgenic SC specific connexin 43 (Cx43) knockout (SCCx43KO) mouse line was used; both knockout (KO) and wildtype (WT) males of different ages from juvenile to adult were compared. The object was to elucidate a possible role of Cx43 on the expression pattern and regulation of occludin. This conditional KO mouse line lacks the gap junction gene Gja1 (coding for Cx43) only in SC and reveals impaired spermatogenesis. The qRT-PCR indicates an increase in occludin mRNA in adult KO mice. These results correspond to the occludin protein synthesis of adult mice. Additionally, during puberty, occludin localization at the BTB barrier in KO mice is delayed. Our study demonstrates spatiotemporal alterations in occludin mRNA- and protein-expression, indicating that Cx43 might act as a regulator for BTB formation (and function).
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    The antiprogestins mifepristone and onapristone reduce cell proliferation in the canine mammary carcinoma cell line CMT-U27
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Guil Luna, Silvia; Hellmén, Eva; Sánchez Céspedes, Raquel; Millán, Yolanda; Martín de las Mulas, Juana
    Canine mammary tumours (CMTs) represent nearly half of all tumours in female dogs and some 50% have malignant behaviour. Simple epithelial carcinomas have shorter disease free periods after surgery and a higher reduction of the proliferation index reduction after antiprogestin aglepristone treatment in vivo related to the expression of progesterone receptors (PR). These findings make simple carcinomas good candidates for endocrine therapy. To further explore this possibility, the effects of the antiprogestins mifepristone (RU486) and onapristone (ZK299) on cell viability and PR expression of the canine mammary carcinoma cell line isolated from a simple epithelial carcinoma CMT-U27 were studied. Twenty five percent of CMT-U27 control cells expressed PR. RU486 (p<0.05) and ZK299 (p<0.05) reduced the number of viable cells (WST-8 test) at 24h but only the latter treatment reduced significantly PR expression in viable tumour cells at 24h of incubation. The results suggest that both RU486 and ZK299 induce a decrease in the number of viable CMT-U27 tumour cells with different effects on PR expression. The canine mammary carcinoma cell line CMT-U27 is sensitive to the effects of antiprogestins and may serve to further explore the role of these drugs in canine mammary carcinomas.
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    Osteopontin expression and localization of Ca2+ deposits in early stages of osteoarthritis in a rat model
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Martínez Calleja, América; Velasquillo, Cristina; Vega López, Marco A.; Arellano Jiménez, M. Josefina; Tsutsumi Fujiyoshi, Victor K.; Mondragón Flores, Ricardo; Kouri Flores, Juan B.
    Calcium deposits have been related to articular cartilage (AC) degeneration and have been observed in late stages of osteoarthritis (OA). However, the role of those deposits, whether they induce the OA pathogenesis or they appear as a consequence of such process, is still unknown. In this work, we present the kinetics of expression and tissue localisation of osteopontin (OPN), a mineralisation biomarker, and calcium deposits in samples from (normal, sham) and osteoarthritic cartilage (in a rat model). Immunohisto-chemical and Western blot assays for OPN, as well as Alizarin red staining for calcium deposits were performed; superficial, middle, and deep zones of AC were analysed. An increased expression of OPN and calcium deposits was found in the osteoarthritic cartilage compared with that of control groups, particularly in the superficial zone of AC in early stages of OA. In addition, the expression and localisation of OPN and calcium deposits during the OA pathogenesis suggest that the pathological AC mineralisation starts in the superficial zone during OA pathogenesis.
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    Detection and characterisation of disseminated tumour cells in bone marrow of breast cancer patients by immunostaining of Her-2 and MUC-1 in combination with Thomsen-Friedenreich (CD176)
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Andergassen, Ulrich; Kölbl, A. C.; Zebisch, M.; Heublein, Sabine; Hutter, S.; Ilmer, M.; Schindlbeck, C.; Friese, K.; Jeschke, U.
    Disseminated tumour cells (DTCs) in the bone marrow derive from many primary tumours, such as breast cancer. Their mere existence hints to present or future metastasis and implicates a worse prognosis for the patient. DTCs may possess different characteristics in comparison to the primary tumour due to events like Epithelial-Mesenchymal-Transition. Therefore these cells might be able to survive chemotherapy and cause relapses of the disease at a later point. We aimed to detect and further characterise DTCs by an immunostaining approach with three different antigen markers (Her-2, MUC-1 and TF, also known as CD176). For that reason, bone marrow of 41 breast cancer patients was obtained during surgery; DTCs were enriched by density gradient centrifugation and cytospins were prepared. After fixation, immunofluorescent double-stainings were carried out with antibodies against CD176 in combination with HER-2 or MUC-1. Cells co-expressing two antigens were found in all staining combinations (Her-2 and CD176: 46.14%; MUC-1 and CD176: 18.15% of all cases). Cells that stained for a single antigen only were also found (Her-2: 36.86%; MUC-1: 34.45%; CD176: 29.65% of all cases). Significant correlations between the stainings of all markers could be shown (p<0,001). In conclusion, Thomsen-Friedenreich Antigen (TF, CD176) is a promising marker in combination with the established marker Her-2 and other markers like MUC-1. These results may serve as a basis for future DTC detection routines and help to individualize medical treatment, reducing side effects and increasing the efficiency of the therapy.