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Browsing by Subject "Immunohistochemistry"

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    A subset of solitary fibrous tumors express nuclear PAX8 and PAX2: a potential diagnostic pitfall
    (Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) McDaniel, Andrew S.; Palanisamy, Nallasivam; Smith, Steven C.; Robinson, Dan R.; Wu, Yi-Mi; Chinnaiyan, Arul M.; McHugh, Jonathan B.; Greenson, Joel K.; Kunju, Lakshmi P.
    Solitary fibrous tumor (SFT), a mesenchymal neoplasm with widespread anatomic distribution, can be diagnostically challenging in limited samples. We recently encountered an aspirate of a pancreatic mass, incorrectly interpreted as metastatic renal cell carcinoma based on strong PAX8 expression by immunohistochemistry (IHC). After resection, morphologic features with additional IHC (CD34 positivity) correctly identified this lesion as a SFT. PAX8 and PAX2 are commonly used as renal tumor markers; however, no series has investigated PAX8 or PAX2 expression in SFT. IHC for PAX8 and PAX2 was performed on 41 SFTs (biopsy and resections) from varying sites. Eight were histologically malignant and eight were recurrences of previous resections. PAX8 staining was observed at least focally in 26.8% (11 of 41) SFT cases; additionally, PAX2 was positive in 12.2% (5 of 41 cases) of SFTs. For PAX8 and PAX2 positive cases 45.6% and 40%, respectively, showed diffuse expression. No correlation was found between PAX8/PAX2 positivity and age, tumor size, site, malignancy, or recurrence. In conclusion, a substantial minority of SFTs express PAX8 and PAX2 via IHC. This presents a diagnostic pitfall when evaluating possible metastases from the kidney, particularly when primary tumors show sarcomatoid or spindle cell morphologies.
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    A useful immunohistochemical approach to evaluate intraductal proliferative lesions of the breast and to predict their prognosis
    (Murcia: F. Hernández, 2011) Omi, Yoko; Yamamoto, Tomoko; Okamoto, Takahiro; Obara, Takao; Kobayashi, Makio
    An examination was performed on 16 intraductal proliferative breast lesions diagnosed as intraductal papillomas (IP) or usual ductal hyperplasia (UDH), which were followed up for more than 3 years. An immunohistochemical marker panel combining myoepithelial markers, high-molecular-weight keratin (HMWK) and neuroendocrine markers was used. Two of 11 IP cases were re-evaluated as atypical ductal hyperplasia (ADH) and ductal carcinoma in situ (DCIS). These cases developed breast cancer after the first operation. One IP case showed repeated recurrences. None of the other IP and UDH cases had breast cancer or recurrence. The ADH, DCIS and the recurrent IP showing a solid growth lacked myoepithelia, but the recurrent IP expressed HMWK, immunohistochemically. Interestingly, these three lesions were weakly positive for neuroendocrine markers. All other IPs and UDHs, including lesions having solid components, were negative for neuroendocrine markers, and most of them were positive for myoepithelial markers and/or HMWK. A combination of the above immunohistochemical markers seems useful to evaluate intraductal proliferative lesions and to predict their prognosis. In particular, intraductal proliferative lesions with solid components exhibiting positivity for neuroendocrine markers should be followed up carefully to monitor breast cancer risk or recurrence.
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    Aberrant nuclear beta-catenin expression in the spindle or corded cells in so-called corded and hyalinized endometrioid carcinomas. Another critical role of the unique morphological feature
    (Murcia : F. Hernández, 2009) Wani, Yoji; Saegusa, Makoto; Notohara, Kenji
    Corded and hyalinized endometrioid carcinoma (CHEC), showing spindle and/or corded (SPICO) cells often in the background of hyalinized stroma, is a rare variant of uterine endometrioid carcinomas. The aim of our study was to explore the status of cell-adhesion molecules (beta-catenin, Ecadherin) in CHECs and to survey whether immunostains for beta-catenin and p53 can help to distinguish CHECs from their morphological mimics: malignant mixed mullerian tumors (MMMTs) and uterine tumors resembling ovarian sex-cord tumors (UTROSCTs). Immunohistochemistry was performed and scored for each element as follows: 0: negative, 1+: <10% positive cells, 2+: 10-30%, 3+: >30%. The SPICO patterns were classified as spindle/fusiform; 3, corded; 1, and both; 2. SPICO components consisted of <10%: 4, 10-30%: 1, >30%: 1. Five contained squamous components. In SPICO elements of all CHECs, nuclear beta-catenin expression (score: 1+; 1, 2+; 2, 3+; 3) and complete loss of membranous expression of E-cadherin was observed. In contrast, comparable components (sarcomatous ones for eight MMMTs or sex-cord-like ones for six UTROSCTs) showed no nuclear positivity for beta-catenin. p53 expression was observed in SPICO (64.7%), sarcomatous (87.5%), and sex-cord-like (50%) components, and sarcomatous areas of most MMMTs notably showed diffuse and intense staining. Sequence analysis of PCR amplification products of exon 3 of the beta-catenin gene revealed mutation in all cases, except two lacking SPICO components represented on microdissected areas. Our results suggest that alterations in beta-catenin/E-cadherin complex play a critical role in SPICO features. Immunostain for beta-catenin and p53 is a promising approach for distinguishing CHECs from MMMTs and UTROSCTs
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    Adrenal cyst with both Müllerian and mesothelial differentiation - a clinicopathological and immunohistochemical study with implications for histogenesis
    (Universidad de Murcia. Departamento de Biología Celular e Histología, 2017) Kopersk, Łukasz i; Szczepankiewicz, Benedykt; Pihowicz, Paweł; Fus, Łukasz; Wolińska, Ewa; Górnicka, Barbara
    True epithelial-lined cysts are rare forms of adrenal cystic lesions, the pathogenesis of which is still not fully understood. In this report we present a case of an adrenal cyst diagnosed incidentally on imaging in a 31-year-old, previously healthy, obese woman. Due to non-specific hormonal disorders and enlargement of the lesion, a right-sided laparoscopic adrenalectomy was performed. The cyst was lined predominantly by ciliated cuboidal-to-columnar, Müllerian-type epithelium, and focally by flat-to-cuboidal, mesothelium-like lining. Immunohistochemistry demonstrated a strong positive reaction in the cells of both types of lining for CKAE1/AE3, CK19, CK7 and WT1, and both had a negative reaction for CK20, CD34, Melan-A, SF1, TTF1, SMA and CDX2. The cells of the ciliated cuboidal-to-columnar epithelium were strongly positive for PAX8, ER, Ep-CAM and EMA, focally positive for PR, and were negative for calretinin, whereas the cells of the flat-to-cuboidal lining were positive for calretinin and podoplanin and showed only a weak positive response in individual cells for PAX8, EMA and EpCAM, but were negative for ER and PR. This is the first reported case of an adrenal ciliated epithelial cyst with Müllerian differentiation (confirmed immunohistochemically) in the English literature. The differences in morphology and immunophenotype of the two types of lining (epithelial Müllerian phenotype versus mesothelial phenotype), suggest that some adrenal epithelial cysts probably form due to metaplasia of mesothelium-derived lining. A similar mechanism may also be involved in the pathogenesis of at least some of the so-called Müllerian cysts (or inclusions) in other locations
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    Adult-onset Alexander disease with a heterozygous D128N GFAP mutation: a pathological study
    (Universidad de Murcia. Departamento de Biología Celular e Histología, 2019) Cabrera Galván, Juan José; Martínez Martin, María Soledad; Déniz García, Daniel; Araujo Ruano, Eduardo; Travieso Aja, María del Mar
    The various forms of Alexander disease (AD) have been linked to heterozygous point mutations in the coding region of the Human glial fibrillary acidic protein (GFAP) gene. The aim of this study was to confirm and characterise an adult variant of AD based on the presence of Rosenthal fibres, which were identified at brain autopsy. We performed histological and immunohistochemical studies and mutation screening by cycle sequencing of exons 1, 4, 6, and 8. A heterozygous D128N GFAP mutation, previously described in three other cases of adult-onset AD (AOAD), was genetically confirmed. The mutation was seemingly sporadic. Symptoms of the female, 65-year-old patient started with occasionally asymmetric motor impairment and concluded, 23 months later, with a lack of spontaneous movement in all four limbs, reduced consciousness, an acute respiratory problem, and eventually lethal exitus. The most striking characteristics were a cerebellar syndrome with subsequent clinical signs due to brainstem and spinal cord involvement. The final diagnosis was based on a complete autopsy, detection of Rosenthal fibres, GFAP, vimentin, alpha B-crystallin, ubiquitin, hsp27, neurofilament, and synaptophysin, and the identification of the corresponding GFAP gene mutation. Blood analyses were positive for ANA and rheumatoid factor. In conclusion, this work describes sporadic, rapidly advancing AOAD in a female patient and links it with other published cases with the same mutation. Reflections are provided on the influence of vasculitis and ANA in AD as well as the presence of Rosenthal fibres in the neurohypophysis.
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    Alendronate effect in esophagus, stomach and liver: An animal based pathological study
    (Universidad de Murcia, Departamento de Biologia Celular e Histiologia, 2020) Papamitsou, Theodora; Sotiriou, Sotiris; Papakoulas, Apostolos; Toskas, Alexandros; Kamperis, Dimitrios; Karachrysafi, Sofia; Dietrich, Eva-Maria; Lialiaris, Stergios; Sioga, Antonia
    Bisphosphonates are commonly used in clinical practice. Their effectiveness is indisputable, however their adverse effects, especially in the GI tract, are still controversial. In our report, we demonstrate pathological findings of the effect of systematic alendronate administration in esophagus, stomach and the liver of an in vivo animal model of 15 Wistar rats. Light microscopy with immunohistochemistry and electron microscopy were used. Microscopic findings of inflammation of the stomach and mild hepatic dysfunction were observed. Conclusively, alendronate can potentially affect gastric mucosa and liver function on this animal experimental model
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    Alterations in diet consistency and variation in Hypoxia Inducible Factor-1α expression in condylar chondrocytes
    (Murcia: F. Hernández, 2011) Kaklamanos, Eleftherios G.; Papamitsou, Theodora; Sioga, Antonia; Economou, Louise
    Objective. To investigate the expression of HIF-1α during the postnatal development of the mandibular condyle under normal and soft consistency diet conditions. Materials and Methods. Forty eight Wistar-Furth rats, aged 21 days, were divided into two groups, each being fed with either normal or soft consistency diet. Three animals from each group were sacrificed after 3, 7, 10 days (initial period), 14, 17 days (intermediate period), and 21, 24 and 28 days (final period) after the start of the experiment. Immediately after sacrifice, the mandible was excised surgically, fixed, demineralised and embedded in paraffin. Six µm thick sections were obtained, processed for conventional and immunohistochemical staining and observed in the optical microscope. HIF-1α expression was assessed semiquantitavely and graded separately for the nucleus stained cells and the cells stained exclusively in the cytoplasm. Differences in HIF-1α expression in the experimental groups were evaluated statistically. Results. HIF-1α expression was evident in the proliferative and chondroblast layers. No differences were observed between the anterior, the intermediate and the posterior parts of the condylar cartilage. In the normal consistency diet group, nuclear HIF-1α expression increased gradually until the end of the experiment. On the contrary, in the soft diet animals, nuclear HIF-1α expression increased only at the final period of the experiment. The normal diet fed animals exhibited more intense nuclear HIF-1α expression compared to cytoplasmic expression. Conclusions. HIF-1α expression in condylar chondrocytes varies under altered conditions of diet consistency
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    Altered expression of urokinase-type plasminogen activator and plasminogen activator inhibitor in high-risk soft tissue sarcomas
    (Murcia : F. Hernández, 2007) Benassi, M.S.; Ponticelli, F.; Azzoni, E.; Gamberi, G.; Pazzaglia, L.; Chiechi, A.; Conti, A.; Spessotto, P.; Scapolan, M.; Pignotti, E.; Bacchini, P.; Picci, P.
    In recent years, classification of soft-tissue sarcomas (STS) has improved with cytogenetic analyses, but their clinical behavior is still not easily predictable. The aim of this study was to detect alterations in the urokinase-type plasminogen system, involved in tumor growth and invasion, by comparing mRNA levels of its components with those of paired normal tissues, and relating them with patient clinical course. Real-time PCR was performed on human STS cell lines and tissues from highly malignant STS, including leiomyosarcomas and malignant fibrous histiocytomas, to evaluate the expression of urokinase-type plasminogen activator (uPA), uPA receptor (uPAR) and plasminogen activator inhibitor-1 (PAI-1). Immunohistochemistry of gene products was also performed. Median mRNA values of all genes studied were higher in tumors than in paired normal tissues. In agreement with data on STS cell lines, significant upregulation for uPA and PAI-1 genes compared to reference values was seen. Moreover, different levels of expression were related to histotype and metastatic phenotype. There was accordance between uPA mRNA and protein expression, while immunodetection of PAI-1 product was weak and scattered. In recent years, classification of soft-tissue sarcomas (STS) has improved with cytogenetic analyses, but their clinical behavior is still not easily predictable. The aim of this study was to detect alterations in the urokinase-type plasminogen system, involved in tumor growth and invasion, by comparing mRNA levels of its components with those of paired normal tissues, and relating them with patient clinical course. Real-time PCR was performed on human STS cell lines and tissues from highly malignant STS, including leiomyosarcomas and malignant fibrous histiocytomas, to evaluate the expression of urokinase-type plasminogen activator (uPA), uPA receptor (uPAR) and plasminogen activator inhibitor-1 (PAI-1). Immunohistochemistry of gene products was also performed. Median mRNA values of all genes studied were higher in tumors than in paired normal tissues. In agreement with data on STS cell lines, significant upregulation for uPA and PAI-1 genes compared to reference values was seen. Moreover, different levels of expression were related to histotype and metastatic phenotype. There was accordance between uPA mRNA and protein expression, while immunodetection of PAI-1 product was weak and scattered.
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    Amelioration of hypercholesterolemiainduced hepatic changes with red grape juice: A histopathological study
    (F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2014) Al-Ahmadi, Ahlam Abdulaziz; Ali, Soad Shaker; Ayuob, Nasra Naeim; Al Ansary, Abeer Khaled
    Objectives: Hypercholesterolemia was confirmed as a risk factor for hepatic fibrosis, as well atherosclerosis and coronary heart disease. This biochemical and histoplathological study was conducted to investigate the possible protective effect of red grape against hepatic injury induced by a high-cholesterol diet (HCD). Material and methods: Thirty male Wister rats were randomly divided into three groups (n=10): the control received saline, the induction group was fed HCD, and the treated group was fed a HCD and 0.4 ml of 100% red grape juice (RGJ) for 13 weeks. After the animals were sacrificed, liver tissue samples were taken to be processed for light and electron microscopy examination. Results: The administration of the RGJ and HCD significantly decreased the animals’ blood glucose, insulin, cholesterol, triglycerides, Low Density Lipoprotein levels and increased their High Density Lipoprotein level compared to the rats fed the HCD alone. It also decreased the periportal (macro- and microvesicular) steatosis, fibrosis, lymphocytic infiltration and blood sinusoidal congestion that were observed in HCD-fed rats alone. The RGJ reduced the number of activated myofibrobasts. This was confirmed by a reduction in the expression of alpha smooth muscle actin and desmin. The RGJ increased, although not significantly, the expression of endothelial Nitric Oxide Synthetase. Conclusion: The administration of RGJ succeeded in alleviating the biochemical and, to some extent, the histopathological changes induced by the high cholesterol diet. Consumption of fresh RGJ or its pharmaceutical preparations is advised especially for those who are used to eat a high fat diet.
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    An effective and practical immunohistochemical protocol for bone specimens characterized by hyaluronidase and pepsin predigestion combined with alkaline phosphatase-mediated chromogenic detection
    (F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2015) Li, Shangfu; Liu, Bin; Tian, Ming; Zhang, Liangming; Tickner, Jennifer; Xu, Jiake; Rong, Limin
    The aim of this study was to provide an effective procedure for immunohistochemistry (IHC) investigations of bone specimens. Samples from rat femoral and human vertebral bone were processed with a detailed and effective IHC protocol summarized here. First, a novel antigen retrieval (AR) method of hyaluronidase combined pepsin predigestion (H+P) was established and the optimal concentration and pH value for AR of bone specimens were determined. Second, the newly developed method was compared with existing AR methods (boiling in sodium citrate, hyaluronidase predigestion (H) and pepsin predigestion (P), with PBS only as the negative control) using two chromogenic detection systems (horseradish peroxidase (HRP) and alkaline phosphatase (AP)) to evaluate their efficacy in obtaining the best IHC results for bone samples. Considering the drawbacks of significant shrinking and detachment from slide for heat retrieval methods and the only moderate immunolabeling for H and P, H+P was the optimal AR method for IHC of bone specimens with the advantages of both good morphological preservation and strong immunoreactivity. Moreover, AP-mediated chromogenic detection was superior to HRP-labeled chromogenic detection due to significantly less nonspecific staining. In conclusion, we presented an effective and practical IHC protocol for bone specimens characterized by H+P predigestion combined with APmediated chromogenic detection. Finally, a detailed troubleshooting guide was provided for common mistakes that occur during IHC processing of the bone tissue samples.
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    An immunohistochemical analysis of the temporal and spatial expression of growth factors FGF 1, 2 and 18, IGF 1 and 2, and TGFß1 during distraction osteogenesis
    (Murcia : F. Hernández, 2007) Haque, T.; Amako, M.; Nakada, S.; Lauzier, D.; Hamdy, R.C.
    Distraction osteogenesis (DO) is a well established surgical technique that generates new bone by gradual distraction of two bony segments. In this study, we investigated the temporal and spatial profile of FGF 1, 2 and 18, IGF 1 and 2, and TGFß1 during distraction osteogenesis using immunohistochemistry. An osteotomy was performed on the right tibia of 13 white male New Zealand rabbits. After a delay of 7 days, distraction was started at a rate of 0.25mm/12hrs for 3 weeks which was followed by a 3 week period of consolidation. Immunohistochemical analysis was performed on a weekly interval to determine the expression of the growth factors. Staining of all growth factors was apparent at various levels in the centre and callus region in fibroblasts and chondrocyte cells. FGF2 however, showed continued high expression in osteoblasts. Within two weeks after the end of distraction all growth factors showed a reduction in expression except for FGF18 which maintained high levels of expression (up to 100% staining) throughout the distraction and consolidation phases. The study suggests that in comparison to the other investigated growth factors, FGF18 may play in important role throughout the entire process of distraction osteogenesis.
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    An immunohistochemical study of the expression of cell-cycle-regulated proteins p53, cyclin D1, RB, p27, Ki67 and MSH2 in gallbladder carcinoma and its precursor lesions
    (Murcia : F. Hernández, 2005) Xuan, Y.H.; Choi, Y.L.; Shin, Y.K.; Kook, M.C.; Chae, S.W.; Park, S.M.; Chae, H.B.; Kim, S.H.
    Gallbladder carcinomas are rare but highly lethal neoplasms. We examined the expression of five cell-cycle-related molecules (p53, RB, cyclin D1, p27, Ki-67), and MSH2, in 46 carcinomas, 14 adenomas, 15 low-grade dysplasias, 9 intestinal metaplasias and 20 normal gallbladder epithelia. The expression of these molecules was altered in gallbladder carcinomas and adenomas. In gallbladder carcinomas we observed increased expression of p53, cyclin D1, Ki-67, and MSH2 together with decreased expression of RB and p27 protein. Aberrant expression of cyclin D1 and reduced expression of RB were noted in adenomas, and expression of cyclin D1 was elevated in low-grade dysplasias. However, there was no change in the levels of these cell-cycle molecules in metaplasia. Expression of p53, p27, Ki-67, and MSH2 was correlated with clinical stage (P<0.05) and there was also a correlation between the expression of Ki-67 and MSH-2 and patient age (P<0.05). These results suggest that altered expression of cell-cycle molecules p53, cyclin D1, RB, p27, and of MSH-2 is involved in the progression of gallbladder carcinomas.
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    An immunohistochemical study on the distribution of endocrine cells in the gastrointestinal tract of the musk shrew, Suncus murinus
    (Murcia : F. Hernández, 1990) Nobuo Kitamura; Junzo Yamada; Tohru Watanabe; Tadayuki Yamashita
    The endocrine cells in the gastrointestinal tract of the musk shrew were studied immunohistochemically. Eleven kinds of endocrine cells, immunoreactive for serotonin, somatostatin, gastrin, cholecistokinin, gastric inhibitory polypeptide, motilin, secretin, neurotensin, pancreatic glucagon, enteroglucagon and bovine pancreatic polypeptide, were revealed. In the stomach, serotonin-, somatostatin-, gastrin-, pancreatic glucagon- and enteroglucagon-immunoreactive cells were detected. The first three types of cells predominated and were more abundant in the pyloric glands than in the other stomach regions. In the small intestine, al1 types of endocrine cells were found, each having different distributions and relative frequencies. In the large intestine, 10 types of endocrine cells except cholecystokininimmunoreactive cells were detected. Serotonin- and bovine pancreatic polypeptide-immunoreactive cells were more numerous in the large intestine than in the small intestine.
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    An immunohistochemical survey of peptide-containing nerves in the copulatory apparatus of several male birds
    (Murcia : F. Hernández, 1993) Kitamura, N.; Yamada, J.; Hayashi, Yoshihiro; Nishida, T.
    Distribution of peptide-containing nerves in male copulatory apparatus as well as rectal and cloacal ganglia were examined immunohistochemically in cock, guinea fowl, duck and muscovy duck. Neurofilament protein-immunoreactive nerves were also examined to investigate general distribution of nerves. Substance P (SP)-, vasoactive intestinal polypeptide (VIP)-, calcitonin gene-related peptide (CGRP)-, and neuropeptide Y (NPY)-immunoreactive (IR) nerves were detected in al1 four species of birds whereas leucine enkephalin (LENK)-IR nerves were detected only in duck and muscovy duck. In the rectal and cloacal ganglia, peptide-IR nerves were detected in varying degrees of abundance. Most noticeable distribution was found in the vascular body, where SP-IR nerve fibres were more abundant in duck and muscovy duck, VIP-IR fibres in muscovy duck and CGRP-IR fibres in duck, than in other regions and other species, respectively. A few peptide-IR nerve fibres were detected in other cloacal regions, although a peculiar pattern in distribution unique to a certain region could be not found. The present results suggest that the peptidecontaining nerves participate in the regulation of tumescence, erection and detumescence of male copulatory apparatus of birds and their regulatory mechanisms are different depending on the species.
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    Analysis of allogenic lymphocytes in rat thymus following sublethal irradiation
    (Murcia : F. Hernández, 1997) Ohba, Y.; Fujikura, Y.; Sawada, T.; Tokuda, N.; Morimatsu, M.; Fukumoto, T.
    The effects of allogeneic lymphocytes on the rat thymus following sublethal irradiation were investigated using immunofluorescence. The recovery of thymus weight following irradiation was delayeú in rats 6 days after receiving lymphocytes compared to controls. Allogeneic cells forming colonies were detected by immunofluorescence in both the cortex and medulla of the host thymus, most frequently on day 15 when an appropriate number (3x10~) was injected. The allogeneic cells detected in the host thymus, presumably T lymphocytes, appeared to disturb thymic reconstitution following irradiation. However, double-immunofluorescence staining revealed that allogeneic cells did not affect the thymic stromal microenvironment. Allogeneic cells may have subsequently affected thyrnic tissue via cytokines. It is important to investigate not only the character of allogeneic cells in the host thymus but also the interactions of donor allogeneic cells, host immature lymphocytes and thymic epithelial cells because of the possibility that these allogeneic cells in the host thymus could prevent the rejection of allogeneic transplants.
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    Analysis of pituitary gonadotropin concentration in blood serum and immunolocalization and immunoexpression of follicle stimulating hormone and luteinising hormone receptors in ovaries of postmenopausal women
    (F. Hernández y Juan F. Madrid. Universidad de Murcia: Departamento de Biología Celular e Histología, 2012) Brodowska, A.; Laszczynska, M.; Brodowski, J.; Masiuk, M.; Starczewski, A.
    The participation of gonadotropins in ovarian carcinogenesis is well known and is supported by studies with inhibition of pituitary gonadotropin secretion, which results in a diminished risk of cancer. However, there are few data on localization and expression of Follicle Stimulating Hormone and Luteinising Hormone Receptors (FSHR and LHR) in ovaries of healthy postmenopausal women, and their correlation with FSH and LH concentration in blood serum is unknown. The aim of our study was to analyze gonadotropin concentration in blood serum and the expression of FSHR and LHR in ovaries of 207 postmenopausal women. Patients included in the study were divided into three groups depending on the number of years since menopause. We analyzed the concentration of FSH and LH in blood serum and the expression of FSHR and LHR in ovaries. Ovaries of postmenopausal women showed numerous morphological changes in the cortex and medulla when compared to the structure of ovaries of women at reproductive age. In all groups of patients clefts in the surface epithelium and epithelial inclusion cysts were found. The concentration of FSH and LH in the blood serum of women studied increased significantly with time from menopause. Significant differences between analyzed menopausal groups were found. The highest FSH and LH concentration in blood serum were found in women with the longest period of time from menopause. Quantitatively similar expression of FSHR and LHR was found in ovarian surface epithelial cells, in epithelial inclusion cysts and in the connective tissue cells of ovarian stroma. The intensity of the immunohisto-chemical reaction decreased with time from menopause and with age
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    Analysis of pRb, p16INK4A proteins and proliferating antigens, PCNA, Ki-67 and MCM5 expression in aggressive fibromatosis (desmoid tumor)
    (Murcia : F. Hernández, 2009) Stalinska, Lilliana; Turant, María; Tosik, Dariusz; Sygut, Jacek; Kulig, Andrzej; Kopczynski, Janusz; Dziki, Adam; Ferenc, Tomasc
    Aggressive fibromatosis (desmoid tumor) is a mesenchymal lesion originating from fascial, aponeurotic and muscular connective tissue. It rarely becomes histologically malignant. In this study we analyzed the cell cycle regulation proteins: pRb, p16, and proliferating antigens: Ki-67, PCNA, MCM5 with immunohistochemical method in archival material derived from 27 extra-abdominal (E-AD), 18 abdominal (AD) and 5 intra-abdominal (I-AD) cases of desmoid tumor. None of the examined cases (n=50) of aggressive fibromatosis was pRb-immunonegative. Heterogeneous expression of pRb was observed in 51.85% (14/27) of Group AD cases and in 5.56% (1/18) of Group E-AD cases; positive expression in 48,15% (13/27) of Group AD cases, in 94.44% (17/18) of Group E-AD cases, and in 100% (5/5) of Group I-AD cases. There were no negative cases for p16 staining in any of the examined groups. The number of heterogeneous cases in individual groups was: 33.33% (9/27) in Group AD, 50% (9/18) in Group E-AD and 40% (2/5) in Group I-AD, and positive cases: 66.67% (18/27), 50% (9/18) and 60% (3/5), respectively. Overexpression of PCNA was noted in 98% (49/50) of cases. The positive staining for Ki-67 protein was noted in 25.93% (7/27) in Group AD, in 16.67% (3/18) in Group E-AD and in 60% (3/5) in Group I-AD. None of the examined cases was immunopositive for MCM5 protein. The noted levels of pRb and p16 expression in desmoid cells reflect their function in cell cycle regulation. Probably the unsettled cell cycle progression, especially in G1 phase, is not the cause of aggressive fibromatosis pathogenesis.
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    Analysis of the expression and localization of tight junction transmembrane proteins, claudin-1, -4, -7, occludin and JAM-A, in human cervical adenocarcinoma
    (Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Akimoto, Taishi; Takasawa, Akira; Murata, Masaki; Kojima, Yui; Takasawa, Kumi; Nojima, Masanori; Aoyama, Tomoyuki; Hiratsuka, Yutaro; Ono, Yusuke; Tanaka, Satoshi; Osanai, Makoto; Hasegawa, Tadashi; Saito, Tsuyoshi; Sawada, Norimasa
    Objective. Tight junction proteins have recently been reported to be useful for distinguishing between neoplastic and non-neoplastic tissues. In this study, we evaluated the expression and localization of tight junction transmembrane proteins in human cervical adenocarcinoma and adenocarcinoma in situ (AIS), and we determined whether their expression patterns could distinguish cervical adenocarcinoma from nonneoplastic cervical glands. Methods. Fifty-five patients with cervical adenocarcinoma or AIS were included in this study. Surgical specimens were immunohistochemically stained for claudin (CLDN) -1, -4, -7, occludin, and JAM-A. Results. Significantly higher expression levels of CLDNs and JAM-A were found in cervical AIS and adenocarcinoma than in non-neoplastic glands. In cervical AIS and adenocarcinoma, localization of CLDN-1 and JAM-A was extended throughout the whole cell membranes, whereas they were predominantly expressed at the most apical cell-cell junction in non-neoplastic glands. ROC curve analysis revealed that immunoreactivities of CLDN-1 or JAM-A successfully distinguished neoplasms from nonneoplastic cervical glands with high specificity (CLDN1, 79.1%; JAM-A, 79.1%) and high sensitivity (CLDN1, 84.1%; JAM-A, 95.5%). Conclusions. As expected, there were immunohistochemical differences between cervical adenocarcinoma and non-neoplastic cervical glands by using antibodies against tight junction transmembrane proteins. These results suggest that CLDN-1 and JAM-A are potential biomarkers for cervical adenocarcinoma.
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    Analysis of the molecular expression profile of non small cell lung carcinoma associated to chronic obstructive pulmonary disease
    (Murcia : F. Hernández, 2009) Garcia-Lujan, Ricardo; Conde-Gallego, Esther; Lopez-Ríos, Fernando; Martin de Nicolas, Jose Luis; Sanchez-Céspedes, Montserrat; García-Quero, Cristina; Echave-Sustaeta, José María; Lopez-Encuentra, Angel
    Chronic obstructive pulmonary disease (COPD) is an independent risk factor to develop lung cancer but there are no different functional clusters of biomarkers between patients with non-small cell lung cancer (NSCLC) with or without COPD. To analyse protein expression, in order to find out whether samples of resected NSCLC from patients with COPD present a different molecular expression. Observational, cohort, concurrent study with sampling since treatment of disease in patients with NSCLC in initial stages (pIApIIB) treated surgically in our hospital between October 1993 and September 1997. The study consisted of the elaboration of tissue arrays with samples from resected tumor, using immunohistochemistry as a study method. Univariate analysis and logistic regression analysis were performed in order to determine molecular markers that showed a differential expression in NSCLC of the patients with COPD. We studied thirty-two proteins in 146 patients. 30% of the patients had COPD. Univariate analysis in patients with COPD showed one molecular marker to be overexpressed and five molecular markers to be underexpressed. Multivariate analysis in patients with COPD identified membranous ß-Catenin as a differential biomarker, which displayed an underexpression, with an Odds Ratio (95% Confidence Interval) of 0.26 (0.07-1.01). A significant lowest expression of membranous ß-catenin was detected in NSCLC of the patients with COPD.
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    Angiosarcomas: histology, immunohistochemistry and molecular insights with implications for differential diagnosis
    (Universidad de Murcia. Departamento de Biología Celular e Histología, 2021) Machado, Isidro; Giner, Francisco; Lavernia, Javier; Cruz, Julia; Traves, Víctor; Requena, Celia; Llombart, Beatriz; López Guerrero, José Antonio; Llombart Bosch, Antonio
    Angiosarcomas (AS) represent a heterogenous group of tumors with variable clinical presentation. AS share an important morphologic and immunohistochemical overlap with other sarcomas, hence the differential diagnosis is challenging, especially in poorly-differentiated tumors. Although molecular studies provide significant clues, especially in the differential diagnosis with other vascular neoplasms, a thorough hematoxylin and eosin analysis remains an essential tool in AS diagnosis. In this review, we discuss pathological and molecular insights with emphasis on implications for differential diagnosis in cutaneous, breast, soft tissue and visceral AS.
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