Identification of cholinergic chemosensory cells in mouse tracheal and laryngeal glandular ducts.

Specialized epithelial cells in the respiratory tract such as solitary chemosensory cells and brush cells sense the luminal content and initiate protective reflexes in response to the detection of potentially harmful substances. The majority of these cells are cholinergic and utilize the canonical taste signal transduction cascade to detect "bitter" substances such as bacterial quorum sensing molecules. Utilizing two different mouse strains reporting expression of choline acetyltransferase (ChAT), the synthesizing enzyme of acetylcholine (ACh), we detected cholinergic cells in the submucosal glands of the murine larynx and trachea. These cells were localized in the ciliated glandular ducts and were neither found in the collecting ducts nor in alveolar or tubular segments of the glands. ChAT expression in tracheal gland ducts was confirmed by in situ hybridization. The cholinergic duct cells expressed the brush cell marker proteins, villin and cytokeratin-18, and were immunoreactive for components of the taste signal transduction cascade (Gα-gustducin, transient receptor potential melastatin-like subtype 5 channel = TRPM5, phospholipase C(ß2)), but not for carbonic anhydrase IV. Furthermore, these cells expressed the bitter taste receptor Tas2r131, as demonstrated utilizing an appropriate reporter mouse strain. Our study identified a previously unrecognized presumptive chemosensory cell type in the duct of the airway submucosal glands that likely utilizes ACh for paracrine signaling. We propose that these cells participate in infection-sensing mechanisms and initiate responses assisting bacterial clearance from the lower airways.

Cholinergic brush cells in the trachea mediate respiratory responses to quorum sensing molecules.

AIM: The airway epithelial surface is constantly exposed to inhaled environmental factors and pathogens. Bitter "tasting" bacterial products such as quorum sensing molecules (QSM) can be detected by solitary chemosensory cells of the upper respiratory tract. Recently, we have shown that tracheal brush cells are cholinergic chemosensory cells affecting the respiration upon stimulation with bitter substances. Here, we explore the hypothesis that tracheal brush cells are capable of detection of bacterial products such as QSM resulting in changes in respiration and in induction of local effects, e.g. regulation of mucociliary clearance. MAIN METHODS: Functional analyses of respiration were performed in the trachea using a newly established model for investigation of respiration in spontaneously breathing anesthetized mice upon isolated tracheal stimulation. Influence of N-3-oxododecanoyl-homoserine lactone (3-OxoC(12)-HSL) on cilia-driven particle transport speed (PTS) in the airways was investigated in acutely excised and submerged mouse tracheae. KEY FINDINGS: 3-OxoC(12)-HSL, a Pseudomonas aeruginosa quorum sensing autoinducer, caused a drop in the respiratory rate 2 min after the application at the mucosal surface. The 3-OxoC(12)-HSL-induced effect on respiration was abolished by inhibition of nicotinic receptors with mecamylamine and by removal of the respiratory epithelium. At the same concentration, 3-OxoC(12)-HSL enhanced significantly PTS on the mucosal surface. SIGNIFICANCE: We conclude that cholinergic airway epithelial cells sense bacterial QSM in the airway lining fluid and communicate this to the CNS via ACh release and nicotinic stimulation of sensory neurons. In addition, QSM enhance PTS.

Cholinergic chemosensory cells in the auditory tube.

The luminal composition of the auditory tube influences its function. The mechanisms involved in the monitoring are currently not known. For the lower respiratory epithelium, such a sentinel role is carried out by cholinergic brush cells. Here, using two different mouse strains expressing eGFP under the control of the promoter of choline acetyltransferase (ChAT), we show the presence of solitary cholinergic villin-positive brush cells also in the mouse auditory tube epithelium. They express the vesicular acetylcholine (ACh) transporter and proteins of the taste transduction pathway such as α-gustducin, phospholipase C beta 2 (PLC(ß2)) and transient receptor potential cation channel subfamily M member 5 (TRPM5). Immunoreactivity for TRPM5 and PLCß2 was found regularly, whereas α-gustducin was absent in approximately 15% of the brush cells. Messenger RNA for the umami taste receptors (TasR), Tas1R1 and 3, and for the bitter receptors, Tas2R105 and Tas2R108, involved in perception of cycloheximide and denatonium were detected in the auditory tube. Using a transgenic mouse that expresses eGFP under the promotor of the nicotinic ACh receptor α3-subunit, we identified cholinoceptive nerve fibers that establish direct contacts to brush cells in the auditory tube. A subpopulation of these fibers displayed also CGRP immunoreactivity. Collectively, we show for the first time the presence of brush cells in the auditory tube. These cells are equipped with all proteins essential for sensing the composition of the luminal microenvironment and for communication of the changes to the CNS via attached sensory nerve fibers.

Colon bleeding from both arteriovenous malformation and inflammatory bowel disease. Case report.

INTRODUCTION: Although endoscopy and angiography have changed the management of lower gastrointestinal bleeding and the majority of patients respond to conservative treatment 10-20% of cases have no recognizable site of hemorrhage. About 10-30% of all patients will require operative intervation. A very rare case of massive lower gastrointestinal bleeding in a young patient who was found to suffer from two causes of gastrointestinal hemorrhage in the same time is reported. The patient had to undergo surgery for the control of bleeding. CASE REPORT: A 23 years old male Greek patient presented to the emergency department of our hospital because of three episodes of hematochezia during the last 10 hours. He was admitted to the surgical department for monitoring of his condition. In the next 10 hours the hematochezia continued and the patient although being transfused with three units of packed red blood cells, started to become unstable with his vital signs affected, having also a syncoptic episode. Emergent colonoscopy could not recognize the site of hemorrhage or any other pathology in the colon, but revealed an intestinal lumen full of blood from the anus to the cecum. It was decided that the patient should undergo operation to stop bleeding. An extensive right hemicolectomy was performed. After that the patient remained stable and showed no signs of hemorrage. The histopathological examination of the specimen showed an arteriovenous malformation but also lesions of the mucosa compatible with early inflammatory bowel disease. CONCLUSIONS: In young patients with massive lower gastrointestinal bleeding of unknown origin, extensive right hemicolectomy provides a good and safe therapeutic choice that will control hemorrhage in most cases with the advantage of lower mortality and morbidity rates compared to subtotal colectomy. Close monitoring of the patient postoperatively is essential.

Muscarinic acetylcholine receptor subtypes expressed by mouse bladder afferent neurons.

Cell bodies of afferent neurons located in lumbosacral dorsal root ganglia (DRG) provide Adelta- and C-fibres to the urinary bladder, reporting bladder wall tension, volume and noxious stimuli. Recent studies suggested an involvement of muscarinic acetylcholine receptors (mAChRs) not only in detrusor contractility but also in modulating afferent function, and this has been linked to the beneficial effects of muscarinic antagonists in the treatment of overactive bladder. Here, we aimed to determine the inventory of mAChR subtypes expressed by bladder afferent neurons in the mouse. Bladder afferent neurons were identified by retrograde neuronal tracing using Fast Blue (FB) or 1, 1'-dioctadecyl-3, 3, 3', 3'-tetramethylindocarbocyanine perchlorhydrate (DiI) injection into the detrusor muscle. DRG L6-S1 were recognized as the major location of bladder afferent perikarya with an additional smaller peak at L1/L2. Retrogradely labelled bladder afferents located in DRG L4-S2 were subjected to immunohistochemistry or to laser-assisted microdissection with subsequent RT-PCR to study expression of mAChRs subtypes M1R-M5R. Immunolabelling for mAChR subtype M2R, validated on DRG from M2R gene-deficient mice, demonstrated this subtype on 35% of FB-labelled bladder afferents. RT-PCR demonstrated expression of subtypes M2R, M3R and M4R, but not of M1R and M5R, in pooled samples (30 section profiles each) of laser microdissected DiI-labelled bladder afferent cell bodies. In conclusion, bladder afferent neurons express different subtypes of mAChRs (M2R, M3R and M4R). Thus, processing of sensory information from the bladder appears to be under direct cholinergic control.

Isolated giant mesenteric fibromatosis (intra-abdominal desmoid tumors). Case report.

A rare case of isolated giant mesenteric fibromatosis is presented. The tumor originated from the fibrous mesenteric tissue. The patient underwent laparotomy because of abdominal discomfort and sub-occlusive symptoms due to the giant mass. Differential diagnosis of mesenteric masses is discussed and the Authors also review the literature concerning this rare disease.

The use of large loop excision of the transformation zone in management problems of cervical intraepithelial neoplasia.

We studied 2 groups of women whose management is controversial: those with cervical intraepithelial neoplasia (CIN) grade 2 or 3 on smear, but only CIN grade 1 or no abnormality on target biopsy (Group 1), and those with persistent CIN grade 1 on smear and up to CIN 1 on biopsy (Group 2). We set out to assess whether large loop excision of the transformation zone (LLETZ) was an acceptable method of treating these 2 groups of women. A review of 100 consecutive patients was undertaken. There were 71 women in Group 1 and 29 women in Group 2. The LLETZ procedures were performed under local analgesia and no immediate problems were encountered. Delayed haemorrhage requiring vaginal packing and admission to hospital occurred in 1 patient. In Group 1, histopathology of the LLETZ biopsies showed CIN 2 or 3 in 29 (40.8%) of the women, CIN 1 in 24 (33.8%) and no CIN in 18 (25.3%), and in Group 2, CIN 2 or 3 was seen in 5 (17.2%) of the women, CIN 1 in 11 (37.9%) and no CIN in 13 (44.8%). At 12 months completed follow-up, 4 patients in Group 1 had recurrent CIN 1 or equivocal CIN 1 and 1 patient from Group 2 had recurrent CIN 1, giving an overall recurrence rate of 5 of the 94 patients who completed follow-up (5%). We concluded that LLETZ was a useful procedure in both groups. In Group 1 the provision of a histological diagnosis on the LLETZ biopsy was a check on the accuracy of the cervical smear report. In Group 2, LLETZ offered the advantage of rapidly returning the smear to normal in most patients, and the diagnosis and treatment of those women who actually had a high-grade lesion.

Secondary malignancies in a child with Hodgkin's disease: T-cell lymphoma and myelodysplastic syndrome evolving into acute nonlymphoblastic leukaemia.

Hodgkin's disease (HD) has been linked to an increase risk of second malignant neoplasms (SMN), especially non-Hodgkin's lymphoma (NHL) and acute nonlymphoblastic leukaemia (ANLL). The mutagenic property of cytotoxic therapy as well as defective immunity have been implicated as playing a major role in the development of SMN in patients previously treated for HD. We report a case of a 14-year-old girl with HD who developed two different second malignancies within a latent period of 28 months following HD diagnosis. The patient presented initially with bilateral cervical and supraclavicular as well as mediastinal and paraaortic lymphadenopathy. She was staged as IIIA, nodular sclerosing type HD, and was given eight alternative cycles of MOPP-ABVD followed by "mantle" field radiotherapy to a total dose of 3.3 Gy plus 0.4 Gy to the upper mediastinum. Within 8 months following the completion of therapy, a period of myelodysplasia and progressive severe immune deficiency, considered as a result of initial treatment, occurred. Eighteen months after HD diagnosis while the patient was continuously neutropenic and heavily immunocompromised, a peripheral T-cell lymphoma of the angiocentric immunoproliferative lesion type (AIL) Grade III, appeared in both lungs within and beyond the radiation field, with no evidence of HD in biopsy specimens. After institution of a new chemotherapy regimen (L17M), a satisfactory response regarding NHL lesions was noted. However, 10 months later the myelodysplastic syndrome (MDS) accompanied by complex chromosomal abnormalities evoluted to frank ANLL with a rapid fatal course. This case supports the hypothesis that combined modality treatment accompanied by severe immunodeficiency may result in the development of multiple second malignancies even within a very short latent period, especially in a subgroup of HD patients who may be particularly increased risk of second cancers.

Environmental pollution and malignant lymphomas: a tentative contribution to geographic pathology.

The present study reports of the geographic distribution of 2,878 cases of uniformly classified malignant lymphomas and of 2,349 similar cases from the literature. Data on histological entities are compared with the geographic and socio-economic characteristics of the regions where they preferentially occur. The results provide three patterns in lymphoma distribution: small cell lymphocytic lymphomas are frequent among populations of European extraction and in farming rather than industrial regions. Lymphomas derived from cells of the germinal center are seen preferentially in so-called "underdeveloped countries", and large cell (immunoblastic) lymphomas appear rather characteristic of industrial areas. Besides genetic influences (rare chronic lymphocytic leukemia (CLL) in Asia as compared to Europe), the results suggest environmental cofactors in the pathogenesis of malignant lymphomas. CLL is more frequent in areas with rather low-dose chronic toxic influences such as from the use of fertilizers and pesticides in farming. Germinal center cell lymphomas tend to occur more frequently in countries with nutritional and hygienic deficiencies with frequent infectious diseases. Large cell high malignancy lymphomas apparently prefer highly industrialized regions with pollution of water supplies by more toxic and immunosuppressive substances.

Persistent active Epstein-Barr virus infection and atypical lymphoproliferation. Report of two cases.

Two cases of what is apparently Epstein-Barr virus (EBV)-associated atypical lymphoproliferation which clinically suggested malignant lymphoma are discussed. Immunohistology revealed polyclonal T- and B-cell proliferation resembling progressive infectious mononucleosis. Giant cell formation and marked plasmacytosis was suggestive of Hodgkin's disease. EBV serology was initially not diagnostic, but during the course of the disease, elevated titers against early antigens and EBV-associated nuclear antigen suggested virus replication. EBV DNA was demonstrated in lymphoid cells of both cases by in situ hybridization. One case progressed to overt monoclonal B-cell malignant lymphoma. Atypical polyclonal lymphoproliferation in persistent active EBV infection may thus be considered a prelymphomatous condition.

Classification and differential diagnosis of 20 problem cases of occidental malignant T-cell lymphomas.

Twenty malignant T-cell lymphomas are described in which the initial histological diagnosis was equivocal. Among lymphoma entities initially falsely interpreted on pure morphological grounds were Hodgkin's disease, lymphoepithelioid cellular lymphoma, immunoblastic lymphoma with epithelioid cells, malignant histiocytosis, histiocytosis X, centroblastic lymphoma, undifferentiated carcinoma, and undifferentiated tumour. Variations in the expected mean survival of initially diagnosed tumours and T-cell lymphomas confirm the need for immunohistological cell typing in lymphoma classification.

Non-Hodgkin's lymphomas: a simplified classification scheme.

1,292 cases of malignant Non-Hodgkin's lymphomas were classified according to the Rappaport and Kiel classifications and to the New Working Formulation for Clinical Usage (NWF). Subsequently, all lymphoma entities were grouped together following the simple criteria of cell size and growth pattern (small cell, large cell, and mixed cellularity; nodular and diffuse growth). Supplementing these morphological criteria, were data from immunological and cytochemical cell marker studies. The accumulated information was correlated to clinical-prognostic parameters. The results show that using above simplified criteria, all Non-Hodgkin's lymphomas can be classified according to a simple scheme which is of clinical relevance. It can further be deduced from all currently used lymphoma classifications, avoids confusion, and offers abbreviated data useful for computer storage.

[Special pathologico-anatomical aspects of AIDS--acquired immune deficiency syndrome]. / Spezielle pathologisch-anatomische Aspekte des AIDS, Acquired Immune Deficiency Syndrome.

Data from morphologic and immunologic follow-up studies are presented of ten patients with AIDS. The sequence of morphologic lesions in lymph nodes proceeds from an initial marked hyperplasia of T- and B-zones (follicular hyperplasia with plasmacytosis and diffuse paracortical hyperplasia) to paracortical atrophy with reactive reticulo-histiocytic hyperplasia and to a late irregular B-cell hyperplasia with effacement of lymph node structure. These changes are accompanied immunocytological by a polyclonal increase in B-Lymphocytes while T-helper cells (TH) and T-suppressor cells (TS) initially exhibit a normal 2:1 ratio, an inversion of the TH/TS ratio is observed in advanced stages. Inversion occurs at the time when paracortical atrophy becomes evident in lymph nodes. Pathogenetically, persistent virus infections and a self-sustaining disturbance of immune regulatory mechanisms are discussed.