Tube Tissue Research Articles

Persistence, Immune Response, and Antigenic Variation of Mycoplasma genitalium in an Experimentally Infected Pig-Tailed Macaque (Macaca nemestrina)

Mycoplasma genitalium is a sexually transmitted pathogen associated with several acute and chronic reproductive tract disease syndromes in men and women. To evaluate the suitability of a pig-tailed macaque model of M. genitalium infection, we inoculated a pilot animal with M. genitalium strain G37 in the uterine cervix and in salpingeal pockets generated by transplanting autologous Fallopian tube tissue subcutaneously. Viable organisms were recovered throughout the 8-week experiment in cervicovaginal specimens and up to 2 weeks postinfection in salpingeal pockets. Humoral and cervicovaginal antibodies reacting to MgpB were induced postinoculation and persisted throughout the infection. The immunodominance of the MgpB adhesin and the accumulation of mgpB sequence diversity previously observed in persistent human infections prompted us to evaluate sequence variation in this animal model. We found that after 8 weeks of infection, sequences within mgpB variable region B were replaced by novel sequences generated by reciprocal recombination with an archived variant sequence located elsewhere on the chromosome. In contrast, mgpB region B of the same inoculum propagated for 8 weeks in vitro remained unchanged. Notably, serum IgG reacted strongly with a recombinant protein spanning MgpB region B of the inoculum, while reactivity to a recombinant protein representing the week 8 variant was reduced, suggesting that antibodies were involved in the clearance of bacteria expressing the original infecting sequence. Together these results suggest that the pig-tailed macaque is a suitable model to study M. genitalium pathogenesis, antibody-mediated selection of antigenic variants in vivo, and immune escape.

Transtubal spread of endometrial carcinoma: correlation of intra-luminal tumour cells with tumour grade, peritoneal fluid cytology, and extra-uterine metastasis

To assess the significance of intra-luminal tumour cells (ILTC) within the fallopian tubes of patients with endometrial carcinoma, with emphasis on high grade histological subtypes. Complete histological examination of fallopian tube tissue was performed in 36 low grade endometrioid adenocarcinomas and in a consecutive series of 226 high grade endometrial malignancies including 92 serous carcinomas, 64 grade 3 endometrioid carcinomas, 26 carcinosarcomas, 25 undifferentiated/dedifferentiated carcinomas, and 19 clear cell carcinomas. The presence of ILTC was correlated with peritoneal fluid cytology, histologically confirmed peritoneal tumour spread, and lymph node metastases. ILTC were identified in 26% and 3% of high and low grade carcinomas, respectively. The presence of ILTC correlated strongly with positive peritoneal fluid cytology and with peritoneal metastasis in high grade tumours (both p < 0.001), and there was also a correlation with lymph node metastasis (p = 0.049). ILTC were more common in serous and undifferentiated carcinomas (>30%) but the differences between the high grade tumour subtypes were not statistically significant. These findings suggest that ILTC associated with high grade endometrial cancers have the capacity to implant and invade the peritoneal cavity. Positive cytology in such cases may be more significant than in low grade tumours. Complete tubal examination may provide additional prognostic information in high grade endometrial carcinoma.

A Study of Persistent Unilateral Middle Ear Effusion Caused by Occult Skull Base Lesions

The goal of this article is to review a series of patients with persistent unilateral middle ear effusion (MEE) and to suggest a more contemporary diagnostic algorithm. The author conducted a retrospective chart review of adults with persistent unilateral MEE and normal findings on physical and nasopharyngoscopic examinations whose MEE was eventually found to be caused by a variety of occult skull base lesions. The study population was made up of 79 patients—52 women and 27 men, aged 21 to 83 (mean: 54.8) at presentation—who had been referred to an academic tertiary care medical center between July 1, 1988, and June 30, 2008. Follow-up ranged from 9 months to 19.5 years (mean: 8.7 yr). Of this group, 50 patients (63.3%) had a malignant tumor, 26 (32.9%) had a benign tumor, and 3 (3.8%) had an internal carotid artery aneurysm. Eustachian tube occlusion had been caused by diffuse invasion in 33 patients (41.8%), by intracranial pathology in 24 (30.4%), and by extracranial-infratemporal lesions in 22 (27.8%). Nasopharyngoscopy cannot identify a variety of rare skull base lesions that cause eustachian tube compression or tissue invasion that ultimately leads to MEE. Therefore, patients with unexplained persistent unilateral MEE should undergo coronal magnetic resonance imaging or computed tomography to look for any intra- or extracranial lesions before undergoing ventilation tube placement.

A Study of Persistent Unilateral Middle Ear Effusion Caused by Occult Skull Base Lesions

The goal of this article is to review a series of patients with persistent unilateral middle ear effusion (MEE) and to suggest a more contemporary diagnostic algorithm. The author conducted a retrospective chart review of adults with persistent unilateral MEE and normal findings on physical and nasopharyngoscopic examinations whose MEE was eventually found to be caused by a variety of occult skull base lesions. The study population was made up of 79 patients-52 women and 27 men, aged 21 to 83 (mean: 54.8) at presentation-who had been referred to an academic tertiary care medical center between July 1, 1988, and June 30, 2008. Follow-up ranged from 9 months to 19.5 years (mean: 8.7 yr). Of this group, 50 patients (63.3%) had a malignant tumor, 26 (32.9%) had a benign tumor, and 3 (3.8%) had an internal carotid artery aneurysm. Eustachian tube occlusion had been caused by diffuse invasion in 33 patients (41.8%), by intracranial pathology in 24 (30.4%), and by extracranial-infratemporal lesions in 22 (27.8%). Nasopharyngoscopy cannot identify a variety of rare skull base lesions that cause eustachian tube compression or tissue invasion that ultimately leads to MEE. Therefore, patients with unexplained persistent unilateral MEE should undergo coronal magnetic resonance imaging or computed tomography to look for any intra- or extracranial lesions before undergoing ventilation tube placement.

Prdm13 Mediates the Balance of Inhibitory and Excitatory Neurons in Somatosensory Circuits

Generating a balanced network of inhibitory and excitatory neurons during development requires precise transcriptional control. In the dorsal spinal cord, Ptf1a, a basic helix-loop-helix (bHLH) transcription activator, maintains this delicate balance by inducing homeodomain (HD) transcription factors such as Pax2 to specify the inhibitory lineage while suppressing HD factors such as Tlx1/3 that specify the excitatory lineage. We uncover the mechanism by which Ptf1a represses excitatory cell fate in the inhibitory lineage. We identify Prdm13 as a direct target of Ptf1a and reveal that Prdm13 actively represses excitatory cell fate by binding to regulatory sequences near the Tlx1 and Tlx3 genes to silence their expression. Prdm13 acts through multiple mechanisms, including interactions with the bHLH factor Ascl1, to repress Ascl1 activation of Tlx3. Thus, Prdm13 is a key component of a highly coordinated transcriptional network that determines the balance of inhibitory versus excitatory neurons in the dorsal spinal cord.

The corona of the daffodil Narcissus bulbocodium shares stamen‐like identity and is distinct from the orthodox floral whorls

The structural homology of the daffodil corona has remained a source of debate throughout the history of botany. Over the years it has been separately referred to as a modified petal stipule, stamen and tepal. Here we provide insights from anatomy and molecular studies to clarify the early developmental stages and position of corona initiation in Narcissus bulbocodium. We demonstrate that the corona initiates as six separate anlagen from hypanthial tissue between the stamens and perianth. Scanning electron microscope images and serial sections demonstrate that corona initiation occurs late in development, after the other floral whorls are fully developed. To define more precisely the identity of the floral structures, daffodil orthologues of the ABC floral organ identity genes were isolated and expression patterns were examined in perianth, stamens, carpel, hypanthial tube and corona tissue. Coupled with in situ hybridisation experiments, these analyses showed that the expression pattern of the C-class gene NbAGAMOUS in the corona is more similar to that of the stamens than that of the tepals. In combination, our results demonstrate that the corona of the daffodil N. bulbocodium exhibits stamen-like identity, develops independently from the orthodox floral whorls and is best interpreted as a late elaboration of the region between the petals and stamens associated with epigyny and the hypanthium.

Immunohistochemical distribution of IGF-1, bFGF and their receptors in decidual, embryonic and tubal human pregnancy tissue

Embryo implantation is a complicated process involving mother and conceptus cells differentiation, proliferation and invasion that are essential for successful pregnancy. Almost every cell in the human body is affected by IGF-1 that is one of the most potent natural activators of cell growth and multiplication, and also a potent inhibitor of the programmed cell death. bFGF, acting via its receptor FGFR1, is one of the factors involved in mediating the angiogenesis, proteolysis and apoptosis during the implantation. OBJECTIVE: To establish pregnancy inducted difference of appearance of bFGF, IGF-1 and their receptors in the embryonic, decidual and tubal tissue. STUDY DESIGN: In this study 14 tubal pregnancy and 10 decidual tissue samples were evaluated immunohistochemically in order to define the distribution of bFGF, FGFR1, IGF-1, IGF-1R. RESULTS: The Mann-Whitney U test was used as appropriate for the evaluation of significant differences. FGFR1 appearance dominated on bFGF in the decidual (z=2.539, p=0.01), tubal (z=2.539, p=0.01) and embryonic (z=2.539, p=0.01) tissue. IGF-1 and IGF-1R appearance in the decidual, tubal and embryonic tissue was not statistically different. It was the same as IGF-1 and IGF-1R expression in gravid endometrium, but in the ectopic implantation site IGF-1R was particularly absent (z=1.935, p=0.05), only mesothelium and some epithelial cells stained. CONCLUSION: IGF-1, IGF-1R, bFGF and FGFR1 are widely appearing growth factors in actively developing and differentiating of the human embryonic tissue during the first trimester. Both endometrial and fallopian tube tissues express more FGFR1 than bFGF that testify the stimulation of compensatory adaptation of the organ during pregnancy. IGF-1 and IGF-1R richly appear in gravid endometrium. IGF-1 is widely distributed in both the mother and the embryo tissues but only some of them are IGF-1R marked in a case of ectopic pregnancy. The deficit of IGF-1R in the fallopian tube might bea result of cell growth restriction and the impaired process of trophoblast invasion.

Interruption of CXCL13-CXCR5 axis increases upper genital tract pathology and activation of NKT cells following chlamydial genital infection.

BackgroundRegulation of immune responses is critical for controlling inflammation and disruption of this process can lead to tissue damage. We reported that CXCL13 was induced in fallopian tube tissue following C. trachomatis infection. Here, we examined the influence of the CXCL13-CXCR5 axis in chlamydial genital infection.Methodology and Principal FindingsDisruption of the CXCL13-CXCR5 axis by injecting anti-CXCL13 Ab to BALB/c mice or using Cxcr5−/− mice increased chronic inflammation in the upper genital tract (UGT; uterine horns and oviducts) after Chlamydia muridarum genital infection (GT). Further studies in Cxcr5−/− mice showed an elevation in bacterial burden in the GT and increased numbers of neutrophils, activated DCs and activated NKT cells early after infection. After resolution, we noted increased fibrosis and the accumulation of a variety of T cells subsets (CD4-IFNγ, CD4-IL-17, CD4-IL-10 & CD8-TNFα) in the oviducts. NKT cell depletion in vitro reduced IL-17α and various cytokines and chemokines, suggesting that activated NKT cells modulate neutrophils and DCs through cytokine/chemokine secretion. Further, chlamydial glycolipids directly activated two distinct types of NKT cell hybridomas in a cell-free CD1d presentation assay and genital infection of Cd1d−/− mice showed reduced oviduct inflammation compared to WT mice. CXCR5 involvement in pathology was also noted using single-nucleotide polymorphism analysis in C. trachomatis infected women attending a sub-fertility clinic. Women who developed tubal pathology after a C. trachomatis infection had a decrease in the frequency of CXCR5 SNP +10950 T>C (rs3922).Conclusions/SignificanceThese experiments indicate that disruption of the CXCL13-CXCR5 axis permits increased activation of NKT cells by type I and type II glycolipids of Chlamydia muridarum and results in UGT pathology potentially through increased numbers of neutrophils and T cell subsets associated with UGT pathology. In addition, CXCR5 appears to contribute to inter-individual differences in human tubal pathology following C. trachomatis infection.

Predictive Factors for Locoregional Control After Salvage Reirradiation of Recurrent Head-and-Neck Cancer

A recent study examined prognostic factors for overall survival (OS) after salvage reirradiation (re-RT) for recurrent head and neck cancer (HNC). Although OS is an important endpoint, uncontrolled local recurrence can lead to tumor bleeding, intractable pain, and suffocation. In the clinic, one often has to make difficult decisions regarding whether patients should undergo re-RT, which can contribute to local control but carries significant morbidity. We thus reviewed our experience to identify factors predictive of locoregional control (LRC) to determine a priori who would benefit from fractionated re-RT. From July 1996 to April 2011, 291 patients with non-metastatic recurrent HN cancer underwent fractionated re-RT. After excluding those with multiple prior RT courses and incomplete documentation, 280 patients were eligible for this analysis. Sites of recurrence were: neck (n = 64), oropharynx (n = 46), nasopharynx (n = 37), oral cavity (n = 34), larynx (n = 28), paranasal sinus (n = 27), parotid (n = 19), hypopharynx (n = 7), and other (n = 18). Median prior dose was 65 Gy and time between RT was 32.7 months. One hundred twenty-one patients (43%) had salvage surgery and 184 (66%) had concurrent chemotherapy. Median re-RT dose was 59.4 Gy. Two hundred thirteen patients (76%) received intensity modulated radiation therapy (IMRT). Actuarial endpoints were assessed with the Kaplan-Meier method and a Cox proportional hazards model was used to determine predictive factors of outcome. Median follow-up was 32.2 months (range 1.8-137.7 months). Median LRC and OS were 16.7 and 17.2 months, respectively. Patients treated with IMRT had superior LRC versus non-IMRT (27 vs. 5.4 months, p < 0.001). Dose ≥50 Gy (p < 0.001), surgery (p = 0.003), nasopharynx subsite (p = 0.06), lack of organ dysfunction (feeding tube, tracheostomy, or soft tissue defect) (p < 0.001), time between RT (p = 0.036), and fewer recurrences (p = 0.04) were also associated with increased LRC on univariate analysis. Charlson index (p = 0.35) and histology (p = 0.14) were not significant predictors of LRC. On multivariate analysis, IMRT (p = 0.005), dose ≥50 Gy (p = 0.009), surgery (p = 0.01), nasopharynx subsite (p = 0.025), and lack of organ dysfunction (p = 0.004) were independently associated with increased LRC. In this large cohort of recurrent HNC patients treated with re-RT, we found that IMRT, higher dose, nasopharynx subsite, lack of organ dysfunction, and surgical resection predicted for longer LRC. As fractionated re-RT requires a demanding course, a shorter hypofractionated palliative RT course should be considered for patients with negative features. A nomogram to help physicians make clinical decisions regarding re-RT is being built and will be presented at the meeting.

Interkinetic nuclear migration during early development of midgut and ureteric epithelia

Interkinetic nuclear migration (INM) is a well-known phenomenon that accompanies progenitor expansion in the vertebrate neural tube and non-neural ectoderm-derived epithelial tissues. In INM, progenitor cell nuclei migrate along the apicobasal axis of the epithelial layer in synchrony with cell cycle progression, resulting in 'pseudostratification'. Although INM has long been considered a general feature of epithelial development, detailed characteristics of INM in the gut and ureteric epithelia are little known. In this study, we observe pseudostratification in the developing midgut and ureteric epithelial progenitors by scanning electron microscopy and assess their cell cycle duration by 5-bromo-2'-deoxyuridine labeling. By applying multi-dimensional scaling, we demonstrate the roundtrip migration of nuclei between the basement membrane and the apical side in the developing midgut. Partial INM has been also shown for the ureteric epithelial nuclei. Our findings reveal INM in gut and ureteric progenitors that is similar to that in ventricular neurogenesis, and suggest that INM is a general strategy for the expansion of epithelial progenitors.

Laser Speckle Flowmetry Method for Measuring Spatial and Temporal Hemodynamic Alterations Throughout Large Microvascular Networks

1) To develop and validate laser speckle flowmetry (LSF) as a quantitative tool for individual microvessel hemodynamics in large networks. 2) To use LSF to determine if structural differences in the dorsal skinfold microcirculation (DSFWC) of C57BL/6 and BALB/c mice impart differential network hemodynamic responses to occlusion. We compared LSF velocity measurements with known/measured velocities in vitro using capillary tube tissue phantoms and in vivo using mouse DSFWCs and cremaster muscles. Hemodynamic changes induced by feed arteriole occlusion were measured using LSF in DSFWCs implanted on C57BL/6 and BALB/c mice. In vitro, we found that the normalized speckle intensity (NSI) versus velocity linear relationship (R(2) ≥ 0.97) did not vary with diameter or hematocrit and can be shifted to meet an expected operating range. In vivo, DSFWC and cremaster muscle preparations (R(2) = 0.92 and 0.95, respectively) demonstrated similar linear relationships between NSI and centerline velocity. Stratification of arterioles into predicted collateral pathways revealed significant differences between C57BL/6 and BALB/c strains in response to feed arteriole occlusion. These data demonstrate the applicability of LSF to intravital microscopy microcirculation preparations for determining both relative and absolute hemodynamics on a network-wide scale while maintaining the resolution of individual microvessels.

147P - Hox Gene Expression in Ovarian Cancer

Ovarian cancer is the leading cause of cancer death among all gynaecological cancers. Its aggressive nature is partly due to genetic heterogeneity and the lack of effective treatments strategies. Standard treatment involves cytoreductive surgery followed by chemotherapy using a platinum-based agent. Although initially, patients response well to this treatment, the majority will relapse and develop recurrent disease, predominantly due to the emergence of platinum resistance. Further understanding of the molecular changes which occur during ovarian ontogenesis and in the development of platinum resistance is essential to design new targeted drugs to improve patient prognosis. HOX gene are a family of homeodomain-containing transcription factors that determine cell and tissue identity in the early embryo and are found to be aberrantly expressed in cancer. HOX gene expression in ovarian cancer of different histological subtypes and in primary ovarian tumours were evaluated here. This is the first comprehensive study of HOX gene expression in a cohort of primary ovarian tumours, including statistical analysis of HOX gene expression profiles along with clinic-pathological data of each patient. HOX genes were found to be profoundly dysregulated in ovarian cancer cell lines and primary ovarian tumours, with very little to no expression found in normal ovarian and fallopian tube tissue. A 5-HOX gene signature which predicts poor overall survival in ovarian cancer patients was identified. Platinum resistant disease displayed an overall higher level of HOX gene expression, significantly HOXB4 and HOXB9. This dysregulated HOX expression reported could therefore act as a set of targets for therapeutic intervention. The novel peptide, HXR9, has been developed to block the interaction between HOX proteins and their co-factor, PBX, and therefore subsequent target gene expression. The efficacy of HXR9 treatment of ovarian cancer cells was explored. HXR9 treatment was shown to induce cell death via apoptosis in ovarian cancer cells shown by an increase in apoptotic cells identified by flow cytometric analysis, and increase in caspase-3 activity and the upregulation of pro-apoptotic gene cFos. Enhanced cell cytotoxicity was observed when combining HXR9 with cisplatin to treat platinum resistant cells, revealing a new therapeutic option for drug resistant disease that should be explored further for potential clinical trial investigation. A limiting factor towards the development of new treatment strategies is the lack of a reliable animal model of ovarian cancer. Common methods used to test new drugs involve in vitro investigation and the use of engineered animal models. However, these models do not represent the true heterogeneity and complexity of human ovarian tumours. Therefore, the use of the chicken chorioallontoic membrane (CAM) as a model of ovarian cancer for the testing of anti-cancer drugs was assessed. Cell lines grafted onto the CAM successfully and developing into micro-tumours. Cell cultured from ascites samples of ovarian cancer patients also grafted, but with a less success rate. Morphological and tumour retardation was detected after treatment with HXR9. This demonstrated the potential of this model to be developed for future personalised drug screening.

Primary culture and immortalization of human fallopian tube secretory epithelial cells

Primary human fallopian tube secretory epithelial cell (FTSEC) cultures are useful for studying normal fallopian tube epithelial biology, as well as for developing models of fallopian tube disease, such as cancer. Because of the limited ability of primary human FTSECs to proliferate in vitro, it is necessary to immortalize them in order to establish a cell line that is suitable for long-term culture and large-scale in vitro experimentation. This protocol describes the isolation of FTSECs from human fallopian tube tissue, conditions for primary FTSEC culture and techniques for establishing immortal FTSEC lines. The entire process, from primary cell isolation to establishment of an immortal cell line, may take up to 2 months. Once established, immortal FTSECs can typically be maintained for at least 30 passages.

A novel taspine analog, HMQ1611, inhibits growth of non-small cell lung cancer by inhibiting angiogenesis

In the present study, we investigated the antitumor activity of HMQ1611, a novel synthetic taspine derivative, in vivo and evaluated associated potential antiangiogenesis mechanisms. The proliferation of A549 cells was examined by WST-1 assay in vitro. Tube formation and lung tissue vessel models were used to observe the antiangiogenic activity of HMQ1611. In addition, vascular enodthelial growth factor (VEGF) secretion and KDR kinase activities were measured by ELISA and the HTRF(®)KinEASE(™)-TK assay. In vivo, the antitumor activity was assessed by implantation of A549 cells in athymic mice. The results showed that HMQ1611 inhibited A549 cell proliferation and VEGF secretion, while it significantly inhibited tube formation and tissue vascularization. Furthermore, HMQ1611 inhibited A549 xenograft tumor growth. In conclusion, the results of our study suggest that HMQ1611 has latent properties for the inhibition of angiogenesis which are involved in its antitumor activity.

Chlamydia trachomatis in fallopian tubes of women undergoing laparoscopy for ectopic pregnancy

To study whether Chlamydia trachomatis is absent or persists in a latent state in the fallopian tube at the time of laparoscopic salpingectomy for tubal ectopic pregnancy (EP). We examined tissue of the fallopian tubes for the presence of C. trachomatis from women who underwent laparoscopic salpingectomy for EP. Presence or absence of C. trachomatis was assessed using both Probe Tec ET (define Tec and ET please) and real-time polymerase chain reaction (PCR) (Ausdiagnositic STD 6 assays) DNA amplification. Fresh tubal tissue from 17 women with histological confirmation of EP was examined in a hospital setting for the presence of C. trachomatis. The presence of C. trachomatis DNA was confirmed by PCR using a commercial test (BD ProbeTec ET System), and a real-time enhanced PCR able to detect few copies of the organism. Chlamydia DNA was detected in 0/16 tubal specimens, and in one case, the PCR analysis was not possible for presence of inhibitors. We did not find any evidence of latent infection of C. trachomatis in the fallopian tube at the time of laparoscopic salpingectomy for EP in our study. Although the numbers are small, our results suggest that EP can be considered a late complication of the tubal damage resulted from a previous acute Chlamydia infection and that EP may not be related to a latent persistence of Chlamydia in the fallopian tube.

Influence of the antifolate drug Methotrexate on the development of murine neural tube defects and genomic instability

Impaired folate metabolism is considered a risk factor for neural tube defects (NTDs). However, the relationship between folate deficiency and the risk of NTDs remains unclear, because experimentally induced dietary folate deficiency is insufficient to cause NTDs in non-mutant mice. Methotrexate (MTX) is a specific folate antagonist that competitively inhibits dihydrofolate reductase (DHFR) activity. The objective of this study was to develop a folate dysmetabolism murine model, and study the development of NTDs and its mechanism. Pregnant mice were injected with different doses of MTX [0, 0.5, 1.0, 3.0, 4.5 and 6.0 mg kg(-1) body weight (b/w) intraperitoneally (i.p.)] on gestational day 7.5 and sacrificed on gestational day 11.5. DHFR activity in embryonic tissues was detected, and folate concentrations were analyzed using LC/MS/MS. Copy number variations (CNVs) in neural tube tissues were detected using array comparative genomic hybridization (aCGH). A dose of MTX 4.5 mg kg(-1) b/w, resulted in the highest incidence of NTDs (31.4%) compared with the other groups, and DHFR activities, 5-MeTHF and 5-FoTHF concentrations in embryonic tissues decreased significantly after MTX injection. Furthermore, we found three high-confidence CNVs on chromosome X using aCGH, which was confirmed by RT-PCR and MassARRAY. These results indicate that MTX could cause a folate-associated dysmetabolism, which is similar to that of dietary folate deficiency in mice. The presence of CNVs in neural tube tissues was associated with the development of NTDs.

Fallopian tube metastases of non‐gynaecological origin: a series of 20 cases emphasizing patterns of involvement including intra‐epithelial spread

To determine the frequency and distribution of Fallopian tube involvement in patients with ovarian metastases of non-gynaecological origin. All Fallopian tube tissue was processed for histological examination in a consecutive series of 31 patients with ovarian metastases of non-gynaecological origin. The most common primary sites were appendix (n = 10) colon (n = 7), stomach (n = 6) and breast (n = 4). Twenty cases (65%) showed at least one type of tubal spread. Mural involvement was most common (14 cases) but serosal, intra-vascular, intra-epithelial and intra-lumenal spread were also identified in 12, 9, 8 and 11 cases respectively. Intra-epithelial involvement was restricted to the fimbrial epithelium and mimicked tubal carcinoma in situ (CIS) architecturally. Pagetoid invasion was noted in two of the cases. The Fallopian tubes are commonly involved in patients who have neoplasms metastatic to the ovaries. Metastases may show a CIS-like pattern of intra-epithelial spread and therefore small serous CIS-type lesions may not represent proof of tubal tumour origin in patients who have high-stage pelvic serous carcinomas. The frequency of intra-lumenal tumour cells supports transtubal spread as a likely mechanism for mucosal involvement by metastatic tumours involving the lower genital tract.

Proliferation and recapitulation of developmental patterning associated with regulative regeneration of the spinal cord neural tube

Developmental patterning during regulative regeneration of the chicken embryo spinal neural tube was characterized by assessing proliferation and the expression of transcription factors specific to neural progenitor and postmitotic neuron populations. One to several segments of the thoracolumbar neural tube were selectively excised unilaterally to initiate regeneration. The capacity for regeneration depended on the stage when ablation was performed and the extent of tissue removed. 20% of surviving embryos exhibited complete regulative regeneration, wherein the missing hemi-neural tube was reconstituted to normal size and morphology. Fate-mapping of proliferative adjacent tissue indicated contributions from the opposite side of the neural tube and potentially from the ipsilateral neural tube rostral and caudal to the lesion. Application of the thymidine analog EdU (5-ethynyl-2′-deoxyuridine) demonstrated a moderate increase in cell proliferation in lesioned relative to control embryos, and quantitative PCR demonstrated a parallel moderate increase in transcription of proliferation-related genes. Mathematical calculation showed that such modest increases are sufficient to account for the amount of regenerated tissue. Within the regenerated neural tube the expression pattern of progenitor-specific transcription factors was recapitulated in the separate advancing ventral and dorsal fronts of regeneration, with no evidence of abnormal mixing of progenitor subpopulations, indicating that graded patterning mechanisms do not require continuity of neural tube tissue along the dorsoventral axis and do not involve a sorting out of committed progenitors. Upon completion of the regeneration process, the pattern of neuron-specific transcription factor expression was essentially normal. Modest deficits in the numbers of transcription factor-defined neuron types were evident in the regenerated tissue, increasing particularly in dorsal neuron types with later lesions. These results confirm the regulative potential of the spinal neural tube and demonstrate a capacity for re-establishing appropriate cellular patterning despite a grossly abnormal morphogenetic situation.

Promoter hypermethylation patterns in fallopian tube epithelium of BRCA1 and BRCA2 germ line mutation carriers

BRCA1/2 germ line mutation carriers have a high risk of developing fallopian tube carcinoma (FTC), thought to occur through different early (p53 signatures) and later (dysplasia, intra-epithelial carcinoma) premalignant stages. Promoter hypermethylation of tumour suppressor genes is known to play a key role in (early) carcinogenesis. However, little is known about methylation in normal and (pre)malignant fallopian tube tissue. We identified 14 areas of p53 accumulation in the fallopian tubes of BRCA mutation carriers. Cells from these areas were harvested together with cells from adjacent benign appearing areas. An age-matched non-BRCA sporadic control group (n=13) and eight sporadic FTCs were included as negative and positive controls respectively. Methylation-specific multiplex ligation-dependent probe amplification was used to assess promoter methylation of 70 tumour suppressor genes in all samples. We observed a gradual increase in methylation from sporadic control tissue (median cumulative methylation index (CMI) 568.19) through normal tissue and from areas of p53 accumulation in BRCA carriers (median CMI 687.54 and 676.72) to FTC (median CMI 780.97). Furthermore, the methylation percentage of many individual tumour suppressor genes differed significantly between these groups, gradually increasing as for CMI. Between areas with and without p53 accumulation in BRCA mutation carriers no significant differences were found. In this paper, we have shown that BRCA mutation carriers display increased methylation of tumour suppressor genes in their non-malignant fallopian tube epithelium, closer to methylation levels in FTC than to normal sporadic tissue. Methylation could, therefore, play an important role in the increased risk of gynaecological malignancies in BRCA mutation carriers.

Prolapso de trompa uterina: reporte de casos y revisión de la literatura

Objetivo: el prolapso de la trompa uterina constituye una complicación quirúrgica poco frecuente y habitualmente ocurre de manera secundaria a la histerectomía vaginal. Dicha patología constituye un reto diagnóstico dado el espectro de lesiones benignas y malignas a nivel del cuello uterino. El presente reporte de casos tiene como objetivo revisar la literatura respecto a la etiología, el diagnóstico y el tratamiento de la entidad.Materiales y métodos: se presentan tres casos de prolapso de trompa uterina, así como una revisión de la literatura, a través de una búsqueda electrónica en las bases de datos Medline vía PubMed, Hinari, SciELO y EMBASE, para recopilar información publicada, tanto en inglés como en español, de los últimos 5 años.Conclusión: el prolapso de trompa uterina poshisterectomía, ya sea vaginal o abdominal, es una complicación quirúrgica anómala. El diagnóstico definitivo se realiza por medio del estudio histopatológico, al evidenciar tejido tubárico asociado a un infiltrado inflamatorio. Para su tratamiento, diferentes técnicas quirúrgicas han sido propuestas con el objetivo de resecar, ya sea parcial o totalmente, la trompa prolapsada con reparo de la apertura vaginal.