Immunochemical Study Research Articles

MUC1 Protein Induces Urokinase-type Plasminogen Activator (uPA) by Forming a Complex with NF-κB p65 Transcription Factor and Binding to the uPA Promoter, Leading to Enhanced Invasiveness of Cancer Cells

Mucin 1 (MUC1) is overexpressed in various human malignant tumors and its expression is correlated with a poor prognosis. MUC1 engages in signal transduction by interacting with receptors for growth and differentiation factors, which contributes to the growth and survival of cancer cells. However, the mechanism by which MUC1 promotes cancer cell invasion remains unclear. Microarray analysis revealed that expression of urokinase-type plasminogen activator (uPA) was elevated in MUC1-overexpressing cells. Furthermore, up- and down-modulation of MUC1 expression was clearly correlated with the change of uPA expression. An immunochemical study showed that the distribution of uPA coincided with that of MUC1 in various human cancer tissues. The MUC1 C-terminal domain (MUC1-CD) was associated with nuclear factor-κB (NF-κB) p65 in MUC1-expressing cells. Chromatin immunoprecipitation (ChIP) assays demonstrated that MUC1-CD existed with NF-κB p65 on the uPA promoter. Luciferase assays indicated that the uPA transcriptional activity was correlated with the level of MUC1 expression and that this MUC1-enhancing effect on the uPA transcription was abolished by introduction of mutations into the NF-κB binding sites on the uPA promoter. These results indicate that formation of the MUC1-CD and NF-κB p65 complex enhanced nuclear translocation of NF-κB p65 and subsequent occupancy of NF-κB binding region on the uPA promoter, leading to elevated transcription of uPA. We also demonstrated that uPA induced by MUC1 enhanced the matrix metalloproteinase (MMP)-2 and -9 activities, and consequently promoted cancer cell invasion. Thus, a MUC1 co-operating NF-κB signaling pathway plays a critical role in cancer cell invasion in MUC1-expressing cells.

Порівняльна характеристика методів епітопного картування антигенів протеїнової природи

Comparative analysis of experimental methods ofepitope mapping of protein antigens has been carried out. The vast majority of known techniques are involved in immunochemical study of the interaction of protein molecules or peptides with antibodies of corresponding specificity. The most effective and widely applicable methodological techniques are those that use synthetic and genetically engineered peptides. Over the past 30 years, these groups of methods have travelled a notable evolutionary path up to the maximum automation and the detection of antigenic determinants of various types (linear and conformational epitopes, and mimotopes). Most of epitope searching algorithms were integrated into a computer program, which greatly facilitates the analysis of experimental data and makes it possible to create spatial models. It is possible to use comparative epitope mapping for solving the applied problems; this less time-consuming method is based on the analysis of competition between different antibodies interactions with the same antigen. The physical method of antigenic structure study is X-ray analysis of antigen-antibody complexes, which may be applied only to crystallizing proteins, and nuclear magnetic resonance.

Binding of the Sialic Acid-binding Lectin, Siglec-9, to the Membrane Mucin, MUC1, Induces Recruitment of β-Catenin and Subsequent Cell Growth

Because MUC1 carries a variety of sialoglycans that are possibly recognized by the siglec family, we examined MUC1-binding siglecs and found that Siglec-9 prominently bound to MUC1. An immunochemical study showed that Siglec-9-positive immune cells were associated with MUC1-positive cells in human colon, pancreas, and breast tumor tissues. We investigated whether or not this interaction has any functional implications for MUC1-expressing cells. When mouse 3T3 fibroblast cells and a human colon cancer cell line, HCT116, stably transfected with MUC1cDNA were ligated with recombinant soluble Siglec-9, β-catenin was recruited to the MUC1 C-terminal domain, which was enhanced on stimulation with soluble Siglec-9 in dose- and time-dependent manners. A co-culture model of MUC1-expressing cells and Siglec-9-expressing cells mimicking the interaction between MUC1-expressing malignant cells, and Siglec-9-expressing immune cells in a tumor microenvironment was designed. Brief co-incubation of Siglec-9-expressing HEK293 cells, but not mock HEK293 cells, with MUC1-expressing cells similarly enhanced the recruitment of β-catenin to the MUC1 C-terminal domain. In addition, treatment of MUC1-expressing cells with neuraminidase almost completely abolished the effect of Siglec-9 on MUC1-mediated signaling. The recruited β-catenin was thereafter transported to the nucleus, leading to cell growth. These findings suggest that Siglec-9 expressed on immune cells may play a role as a potential counterreceptor for MUC1 and that this signaling may be another MUC1-mediated pathway and function in parallel with a growth factor-dependent pathway.

Normal Blood and Bone Marrow Wt1 Expression Level in Extramedullary Relapse in AML Patient

AbstractAbstract 4825Wilms' tumor gene 1 (WT1) that encodes a transcription factor has elevated expression in 90% of AML. High level of WT1 gene expression in bone marrow and peripheral blood of patients with AML indicates the presence of leukemia tumor burden. Thus, quantifying assessment of the level of WT1 could predict relapse. It is important to monitor a universal marker in the absence of target molecular genetic abnormalities in the disease onset. There are few data related to extramedullary relapses. Here we describe the patient with extramedullary relapse without concomitant elevation of WT1 expression.The patient is 22 year old male with the first early bone marrow and CNS relapse of AML. Cytogenetics- (45XY, t (3, 5) (q29; q15), −7, der (7), add (11) (p15)). Blast cells immunophenotype was CD45 + CD34 + CD38 + CD117 +. There was high blast count with high expression of WT1 level (Tabl 1). And there was no expression of other molecular genetic prognostic markers. Therapy with high dose of cytarabine with intrathecal administration of cytotoxic drugs was conducted. The complete hematological and cytogenetic remission was achieved after the first induction. Also there was no any blast cell in the CSF, WT1 expression decreased by 1,87 log (WT1 level became normal). Consolidation of remission with three similar courses with high doses of cytarabine was conducted. Extramedullary relapse during bone marrow and CNS remission with unilateral lesion of the lacrimal gland and the periorbital area( 2.8 × 3.4×4.6 cm )was diagnosed in 6 month. Relapse was confirmed by histological and immunochemical study of the left lacrimal gland biopsy, blasts phenotype was similar to the phenotype of bone marrow blasts at AML onset. WT1 level in bone marrow and peripheral blood remained within normal limits: 15.5 WT1/104 ABL copies and 13.4 WT1/104 ABL copies respectively. Therapy with high dose cytarabine and mitoxantrone was used as an induction. After this therapy a local remission was diagnosed according to MRI. Consolidation courses were followed by a course of radiotherapy to the area of the left orbit. Bone marrow remission and low-level expression of WT1 remained to be normal. Thereafter the patient has undergone unrelated allogeneic bone marrow transplantation. Conclusion.We have described isolated AML relapse without elevation of WT1. A possible reason for normal level of expression of WT1 in patients with extramedullary relapse may be a small amount of the extramedullary formation, its location and features of blood supply of the body area.Table 1Blast count in bone marrowWT1/104 ABL copies in bone marrowAt the diagnosis58%3560,6After the induction1%50,8After the first consolidation course1,4%26,7After the second consolidation course0,4%9,6After the third consolidation course0,4%3,04Follow-up2,4%14,3Follow-up1,4%17,7Extramedullary relapse1,4%15,5Day 14 of ≪HDAC≫3,4%29,4After ≪HiDAC≫6%49,2Follow-up before radiotherapy1,6%15Follow-up after radiotherapy32 [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Human DNA damage by the synergistic action of 4‐aminobiphenyl and nitric oxide: An immunochemical study

4-Aminobiphenyl (4-ABP), an aromatic amine is a major environmental carcinogen found mainly in cigarette smoke. It has been vastly implicated in mutagenesis and cancer development. In this study, commercially available human placental DNA was exposed to 4-ABP (1.3 mM) in presence of sodium nitroprusside (SNP; 8 mM) at 37°C for 3 h. The 4-ABP + SNP-mediated structural changes in human DNA were studied by ultraviolet, circular dichroism and fluorescence spectroscopy, thermal melting profile, agarose gel electrophoresis, and nuclease S1 digestibility assay. Spectroscopical analysis and melting temperature studies suggest structural perturbations in the DNA as a result of modification. This might be due to generation of single-stranded regions and destabilization of hydrogen bonds. Modification was also visualized in agarose gel electrophoresis. Furthermore, nuclease S1 digestibility confirmed the generation of single strand breaks. Rabbits challenged with 4-ABP-SNP-modified human DNA-induced high-titer immunogen-specific antibodies, which showed Cross-reaction with modified/unmodified DNA bases and ss-DNA in competitive inhibition assay. The immunogen specificity of induced antibodies against 4-ABP-SNP-modified human DNA was further confirmed in gel retardation assay. It may be concluded that induction of anti-modified DNA antibodies could be due to perturbation in the DNA structure and its subsequent recognition by immunoregulatory cells as a foreign molecule.

Primary breast diffuse large B-cell lymphoma shows an activated B-cell–like phenotype

Patients with diffuse large B-cell lymphoma (DLBCL) can be divided into germinal center B-cell-like subtype (GCB-DLBCL) and activated B-cell-like type (ie non-GCB-DLBCL) and with a prognostic value based on the cell of origin of the tumor as determined by microarray analysis, as well as by immunostain algorithms. To stratify primary breast DLBCL, according to immunostain algorithms of Hans, Choi, and Meyer (Tally), the reason why primary breast DLBCL possesses a poor clinical outcome was further elucidated. Twenty-two cases of primary breast DLBCL, diagnosed according to the 2008 World Health Organization classification of tumors of hematopoietic and lymphoid tissues, were retrospectively studied. Immunochemical study was performed, and follow-up information was analyzed. The histopathologic and immunophenotypic features of all cases were summarized. Among these 22 cases, most (17, 16, and 17, respectively) were assigned to the activated B-cell-like subtypes, few (5, 6, and 5, respectively) were classified as GCB-DLBCL subtype. Fourteen of these 22 cases expressed high proliferative activity (≥40% Ki-67 labeling). Most primary breast DLBCLs have activated B-cell-like subtype characteristics and high proliferative activity; these features might be a significant factor.

Rheumatic masks of plasma cell dyscrasias

Система комплексной поддержки и сопровождения научного журнала Elpub позволяет запустить двуязычный сайт журнала со встроенной системой электронной редакции в соответствии с лучшими издательскими практиками, а так же предусматривает техническую и методическую поддержку со стороны специалистов НЭИКОН.

The diagnostic value of Ki-67, P53 and P63 in distinguishing partial Hydatidiform mole from hydropic abortion

The aim of the present study was to investigate the expression of Ki-67, P53 and P63 in hydropic abortion and partial mole and to explore possible role of these three parameters in distinguishing partial mole from hydropic. The hydropic abortion (n = 13) and partial mole (n = 23) were retrieved from the files of 2002 to 2010 at the Department of Pathology, Peking University People's Hospital and Department of Pathology, Haidian Maternal and Child Health Hospita1. All cases had typical histological features and complete clinical information. All pathologic diagnoses were reviewed by two gynaecologic pathologists and had no conflict. All cases showed P57 negative, which can exclude the diagnosis of CHM. An immunochemical study of the expressions of Ki-67, P53 and P63 was performed. Microscopically, the pathologic characteristics of a HA include villous oedema with minimal to no cistern formation and mild trophoblastic hyperplasia. In contrast, the pathologic features of PHM were characterized by focal trophoblastic hyperplasia and a variable degree of hydropic swelling with central cistern formation. The Ki-67 expression was observed in the nuclear of cytotrophoblastic cell and intermediate trophoblasts population within placental tissue. The Ki-67 expression in HA is less than that in PHM. The p53 expression was observed in the nuclear of cytotrophoblastic cell and intermediate trophoblasts population within placental tissue. There was a significant difference between HA and PHM. Positive staining for P63 was localized in the nuclei of cytotrophoblastic cell population. No significant difference was observed in positive rate of p63 expression between HA and PHM. The high Ki-67 labeling index and over-expression of p53, detected by immunohistochemistry, could serve as useful adjuncts to conventional methods of diagnosis in distinguishing PHM from HA. Due to, however, the limited samples, it needs to expand the number of the cases to verify this conclusion.

A case of monocular blindness as the initial presentation of hepatocellular carcinoma with skull metastasis.

A 52-year-old man suffering from monocular blindness, with light perception only, was admitted to our hospital. The symptom had begun as low vision and developed rapidly within 3weeks into monocular blindness in the right eye, with no other systemic manifestations. Imaging examinations revealed multiple hepatocellular carcinomas in the cirrhotic liver, and tumors at the skull base and vertebra. A pathological and immunochemical study of specimens obtained by endoscopic transnasal tumor biopsy and laminectomy revealed them to be metastatic hepatocellular carcinomas (HCCs). Although the patient underwent radiation therapy and chemotherapy, he died 5months after admission to our hospital. The cranial HCC, involving only the optic canal, may have disturbed the optic nerve in preference to the other cranial nerves. This is the first report of a HCC patient with monocular blindness as the initial presentation of the disease.

Study of Dj-1 protein in tissue specimens, cultured cells and serum of prostate cancer patients

Two isoforms of Dj-1 protein were identified using a proteomic study in tissue specimens from two groups of patients with confirmed benign prostate hyperplasia (BPH) and prostate cancer (PCa). Dj-1 was also found in the cell lines PC-3, DU-145, LNCaP, BPH-1, and the lowest level of Dj-1 was found in BPH-1. Immunochemical study (ELISA) of serum levels of Dj-1, Bcl-2, IGF-1 and IGFBP-3 proteins revealed statistically significant distinctions between two groups of patients (p=0,004, Mann-Whitney test) only for Dj-1. Taken together, these data suggest that Dj-1 protein is a perspective biomarker candidate for PCa.

Mechanisms of Apoptosis on Human Lens Epithelium after Ultraviolet Light Exposure

PurposeThe purpose of this study is to understand the mechanism of apoptosis occurring on a cultured human lens epithelial cell line after exposure to ultraviolet (UV) light. We intended to confirm the presence of cellular toxicity and apoptosis and to reveal the roles of p53, caspase 3 and NOXA in these processes.MethodsCells were irradiated with an ultraviolet lamp. Cellular toxicity was measured by a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Hoechst staining and fluorescent anti-caspase 3 antibodies were used for apoptosis investigation. The quantities of p53, caspase 3, and NOXA were measured by Western blotting for to investigate the apoptosis pathway.ResultsCellular toxicity on the human lens epithelium markedly increased with time after UV exposure. On Hoechst staining, we found that apoptosis also remarkably increased after exposure to ultraviolet light, compared with a control group. In the immunochemical study using anti-caspase 3 antibodies, active caspase 3 significantly increased after exposure to ultraviolet light. On Western blotting, p53 decreased, while caspase 3 and NOXA increased.ConclusionsExposure of cultured human lens epithelial cell lines to ultraviolet light induces apoptosis, which promotes the expression of NOXA and caspase 3 increases without increasing p53. This may suggest that UV induced apoptosis is caused by a p53-independent pathway in human lens epithelial cells.

Stress-related proteins compared in diapause and in activated, anoxic encysted embryos of the animal extremophile, Artemia franciscana

Previous work indicated similarities between diapause and anoxic quiescence in encysted embryos (cysts) of the brine shrimp Artemia franciscana. That possibility was examined further in the present study through an immunochemical study of the following stress-related proteins in low speed supernatants and pellets: hsc70, artemin, p26, hsp21, LEA Group 1 protein and p8. Changes in the amounts and locations of these proteins occurred during the initial period after release of diapause cysts from females, and after activated (diapause-terminated) cysts were made anoxic. However, with the passage of incubation time the patterns seen in both kinds of cysts were more similar than different, lending further support to the possibility that activated anoxic embryos retain many of the mechanisms operative in the previous diapause condition.

Promoting regeneration of peripheral nerves in-vivo using new PCL-NGF/Tirofiban nerve conduits

Poly(ε-caprolactone) (PCL) scaffolds were modified by grafting nerve growth factor (NGF) and Tirofiban (TF), a clinical anti-thrombosis drug, as a new biomaterial for producing nerve conduits to promote the regeneration of sciatic nerves. The successful grafting of NGF and TF onto PCL scaffolds was confirmed by FTIR and ESCA spectra. In-vitro growths of the PC12 cells in PCL-NGF and PCL-NGF/TF scaffolds, determined by MTS, were significantly higher ( P < 0.05, n = 4) than those in the PCL scaffolds following three days of cultivation. Interestingly, this study evaluation of the PCL, PCL-NGF, and PCL-NGF/TF nerve conduits in a 12 mm long gap of the rat sciatic nerve defect model that the gastrocnemius muscle mass of the tested rats in the PCL-NGF/TF groups significantly exceeded those in the PCL-NGF and PCL group. In the rats that had been implanted with PCL-NGF/TF conduits, the generated nerves passed through those conduits, expressing beta-III tubulin (TB), growth association protein-43 (GAP-43) and myelin basic protein (MBP) along their longitudinal axis, and the proximal and distal nerve ends of the rats were successfully connected. Those that had been implanted with PCL and PCL-NGF conduits did not exhibit these effects, as revealed by an immunochemical study of the expressions of the proteins in the conduits. Moreover, counting within the dorsal horn of the spinal cord (C 5) demonstrated that the numbers of CTB-HRP-labeled neurons in the rats that had been implanted with PCL-NGF/TF conduits were significantly higher than those in the other groups. In this study , in-vivo examinations of the use of newly designed PCL-NGF/TF conduits to promote the generation of nerves in a defective rat model significantly increased the gastrocnemius muscle mass, and led to the successful regeneration of nerves that bridged a 12 mm long defected gap of nerves in rats. However, more rats must be tested to confirm the efficacy the newly designed nerve conduits.

Deficiency in the Serum-Derived Hyaluronan-Associated Protein-Hyaluronan Complex Enhances Airway Hyperresponsiveness in a Murine Model of Asthma

Background: Serum-derived hyaluronan (HA)-associated proteins (SHAPs), the heavy chains of inter-α-trypsin inhibitor, covalently bind to HA to form the SHAP-HA complex. The SHAP-HA complex is involved in the pathophysiology of inflammatory diseases, including rheumatoid arthritis. We investigated whether this complex is also involved in airway allergy. Methods: SHAP-HA-deficient (bikunin knockout, KO) mice and wild-type (WT) mice were immunized twice by intraperitoneal injection of ovalbumin (OVA) and exposed to aerosol OVA for 30 min each day for 2 weeks. Twenty-four hours after the final OVA challenge, airway responsiveness to inhaled methacholine (MCh) was measured, and analysis of bronchoalveolar lavage fluid (BALF) and lung histological studies were done. Results: Compared to WT mice, KO mice showed higher airway hyperresponsiveness to inhaled MCh and higher late-phase responses to OVA whereas the early-phase responses were similar. Cell differentials of BALF showed an increased number of macrophages and neutrophils in KO mice. Furthermore, decreased concentrations of soluble tumor necrosis factor receptor-1 (sTNFR1) were found in BALF from KO mice whereas the levels of Th1 and Th2 cytokines were not different from WT mice. Immunochemical study of the lung tissues revealed stronger staining of sTNFR1 in KO than in WT mice. Conclusions: Our results suggest that in this murine asthma model, the SHAP-HA complex has an inhibitory role in the development of airway hyperresponsiveness and allergic airway inflammation which may be attributed, at least in part, to negative feedback mechanisms exerted by sTNFR1, the shedding of which from the cell surface might also be promoted by the SHAP-HA complex.

CLINICAL MANIFESTATIONS OF VASCULITIS AT THE ONSET OF MULTIPLE MYELOMA

The paper describes two patients (a 50-year woman and a 72-year man) in whom the clinical manifestations of ulceronecrotic vasculitis had long preceded before the diagnosis of multiple myeloma was made. In the former, monoclonal cryoglobulinemia type I induced ulcerative lesions with the development of dry toe gangrene and paraproteinemic renal lesion. In the latter, ulceronecrotic vasculitis with the development of dry toe gangrene was a manifestation of monoclonal paraproteinemia without signs of cryoglobulinemia. Both patients were found to have monoclonal blood secretion (РIgGλ и РIgGκ) and urine Bence Jones protein (BJλ+BJκ, BJκ) in the absence of immunological markers of vasculitis developing in patients with rheumatic diseases. Immunochemical study of serum/urine and, when monoclonal secretion of PIg+BJ is detected, further examination for plasma cell dyscrasia should be performed in all cases of vascular disorders (cold allergy, Raynaud's syndrome, purpura, ulcers of cruses, and gangrene of distal phalanxes of the hands/feet) in patients with atypical vasculitis. The timely immunochemical study of blood and urine will make it possible to diagnose plasma cell dyscrasia (different types of myelomic disease, Waldenstrцm macroglobulinemia, primary amyloidosis) at the early stage of the disease and to rule out unjustifiably diagnosed vasculitis in patients with atypical vascular lesions.

Boron deficiency results in induction of pathogenesis-related proteins from the PR-10 family during the legume–rhizobia interaction

Boron (B) deficiency has a strong effect on molecular and cellular plant–bacteria interactions during the development of the legume–rhizobia symbiosis, leading to reduced infection and early necrosis of nodules, resembling a pathogenic-like rather than a symbiotic interaction. Therefore, induction of pathogenesis-related (PRs) proteins was investigated here in legume root nodules. Following two-dimensional electrophoresis and MALDI-TOF spectrometry analysis of proteins extracted from Pisum sativum B-sufficient (+B) or B-deficient (–B) root nodules, two proteins from the family PR10, ABR17 and PR10.1, were identified as highly induced in –B nodules. Analysis of gene expression and the use of anti-ABR17 confirmed that induction occurred in B-deficient young nodules and increased during nodule development. ABR17 was also induced in –B nodules of Phaseolus vulgaris. Boron deficiency did not significantly increase the expression of these PR10 in uninfected plant tissues. Moreover, independent of B, induction was detected in senescent tissues, although at a level weaker than in −B nodules. The immunochemical study of ABR17 antigen distribution showed that it was localized in all tissues of poorly invaded B-deficient nodules and accumulated around bacteria, which showed advanced degradation. These results suggest that, under B deficiency, the rhizobia–legume dialogue fails and the bacterium is recognized as a pathogen by the plant, which reacts to prevent infection by inducing at least these two identified PR10 proteins.

Immunochemical study of protein profiles of Taleghan, Fars, and Lorestan strains of Mycoplasma agalactiae

Agalactia is a contagious disease in sheep and goats. To prevent the disease, a vaccine containing three strains of Mycoplasma agalactiae namely Taleghan (T), Fars (F), and Lorestan (L) is currently used in Iran. In this study, the protein profiles of T, F, and L strains using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with 8.5% gel and Western blotting were studied. Antigens were prepared from these strains, and the amounts of protein were measured using Lowry method9. Protein profiles of the three strains studied by SDS-PAGE revealed only minor differences with as many as 8, 14, and 10 bands detected in T, F, and L strains, respectively. Majority of the proteins were shared by all of the three strains. Western blotting with rabbit antisera against T, F, and L strains detected nine, nine, and eight immunodominant antigenic bands. All of the above strains revealed more or less similar protein profiles with major common immunodominant antigens at 117, 110, 98, 80, 50, and 5 KDa positions.

MIB-1 (Ki-67), Estrogen Receptor, Progesterone Receptor, and p53 Expression in Atypical Cells in Uterine Symplastic Leiomyomas

Nine cases of symplastic uterine leiomyomas were chosen from our files using strict criteria. An immunochemical study of the expressions of Ki-67, estrogen receptor, progesterone receptor, and p53 was performed. For each antigen, the percentages of positive atypical (bizarre) cells and nonatypical cells were compared. Both groups had identical and fairly high estrogen receptor and progesterone receptor immunohistochemical profiles and very low levels of p53 expression. The atypical cells had significantly higher percentages of cells expressing Ki-67 compared with nonatypical cells (17+/-20% vs. 3+/-3.9%). Therefore, rather than being degenerative in nature, the atypical cells are actively proliferating.

Sclerosing Sweat Duct-Like Carcinoma of the Tongue-A Case Report and a Review of the Literature

In this article, we present the second case of a tongue carcinoma with distinct morphological similarities to a primary sclerosing sweat duct carcinoma of the skin. The tumor infiltrated deeply into the striated muscle and exhibited extensive peri- and intraneural growth. Neoplastic cells had mildly pleomorphic nuclei and displayed in addition to small cysts, distinct duct formation with a bilayered epithelium. Mitotic figures were scarce. Despite extensive sampling, no precursor lesion-dysplasia-was found. The immunochemical study showed positive reaction for carcinoembryonic antigen, epithelial membrane antigen, cytokeratin (CK)7, low-molecular weight CK, BerEp4, high-molecular weight CK, CK5, and CK18. p63 was positive in the peripheral parts of the neoplastic ducts with negative reaction in the luminal cells.

Re-innervation pattern of the ‘neovagina’ created from the bladder flap in patients with Mayer-Rokitanski-Kistner-Hauser syndrome: An immunochemical study

Background. Cystovaginoplasty (CVP) is a method of vaginal reconstruction in women with Mayer-Rokitansky-Kistner-Hauser Syndrome (MRKHS). The neo-vagina allows normal sexual intercourses, but after CVP, the sexual life of patients with MRKHS does not differ significantly from normal females. Therefore, we decided to elucidate the pattern of sensory re-innervation of the bladder flap used for the surgery.Methods. Biopsies were taken from vaginal vestibule and urinary bladder during the CVP and 1 year later in four patients with MRKHS. The following neurotransmitters were studied calcitonin gene-related peptide (CGRP), galanin (GAL), vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP).Results. CGRP and PACAP nerve fibres were sparse under the urothelium and in submucosal layer of the neovagina, they were more numerous around blood vessels and in the vicinity of smooth muscles. This was similar to the pattern observed in the urinary bladder. VIP- and GAL-positive nerve fibres were most numerous in the submucosa around blood vessels and in smooth muscle bundles of neovagina. They were distinctly less numerous beneath the epithelium. This innervation pattern mimicked that seen in normal vagina and in vaginal vestibule of patients with MRKHS.Conclusions. Our findings demonstrate considerable nervous system plasticity in the bladder flap. Distribution of presumably sensory CGRP and PACAP immunoreactive nerve fibers was similar to the pattern observed within the intact bladder wall, and VIP or GAL immunoreactive nerve fibers (vasomotor functions) were distributed in a manner similar to that observed in the intact vaginal wall.