A Plant Proteinase Inhibitor from Enterolobium contortisiliquum Attenuates Pulmonary Mechanics, Inflammation and Remodeling Induced by Elastase in Mice.

Proteinase inhibitors have been associated with anti-inflammatory and antioxidant activities and may represent a potential therapeutic treatment for emphysema. Our aim was to evaluate the effects of a plant Kunitz proteinase inhibitor, Enterolobium contortisiliquum trypsin inhibitor (EcTI), on several aspects of experimental elastase-induced pulmonary inflammation in mice. C57/Bl6 mice were intratracheally administered elastase (ELA) or saline (SAL) and were treated intraperitoneally with EcTI (ELA-EcTI, SAL-EcTI) on days 1, 14 and 21. On day 28, pulmonary mechanics, exhaled nitric oxide (ENO) and number leucocytes in the bronchoalveolar lavage fluid (BALF) were evaluated. Subsequently, lung immunohistochemical staining was submitted to morphometry. EcTI treatment reduced responses of the mechanical respiratory system, number of cells in the BALF, and reduced tumor necrosis factor-α (TNF-α), matrix metalloproteinase-9 (MMP-9), matrix metalloproteinase-12 (MMP-12), tissue inhibitor of matrix metalloproteinase (TIMP-1), endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS)-positive cells and volume proportion of isoprostane, collagen and elastic fibers in the airways and alveolar walls compared with the ELA group. EcTI treatment reduced elastase induced pulmonary inflammation, remodeling, oxidative stress and mechanical alterations, suggesting that this inhibitor may be a potential therapeutic tool for chronic obstructive pulmonary disease (COPD) management.

Clinical significance of cellular and acellular mucin pools in rectal carcinoma following preoperative chemoradiotherapy.

BACKGROUND AND OBJECTIVES: The standard treatment for locally advanced rectal cancer (LARC) is neoadjuvant chemoradiotherapy (CRT) followed by surgery. Pathological findings remain the most significant prognostic factor. The presence of mucin pools and their prognostic significance is a controversial issue. The aim of this study was to analyze the incidence of cellular and acellular mucin pools and their clinical significance. METHODS: Four-hundred and forty-six consecutive prospectively collected specimens from patients with LARC treated with long-course preoperative CRT and surgery were analyzed. Kaplan-Meier analysis was performed. RESULTS: Mucin pools were present in 182 specimens (40.8 %); 66 (14.7 %) were acellular, and viable tumor cells were identified in 116 (26 %). The complete pathological response rate was 13.5 % (60 of 446). With a median follow-up of 79.0 months, the 5- and 10-year disease-free survivals for patients with acellular and cellular mucin pools were 81.5, 78.1, 63.7 and 61.2 %, respectively (p ≤ 0.026). The presence of cells in the colloid response to treatment was associated with a 17.8 and 16.9 % decrease in 5- and 10-year disease survival vs. acellular colloid response. CONCLUSIONS: Our results suggest that cellular mucin pools are an indicator of an aggressive phenotype and harbingers of a worse prognosis.

Mucinas salivales: estructura química, mecanismos de liberación y participación en la defensa no inmunológica de la cavidad bucal / Salivary mucins: chemical structure, release mechanisms and participation in the non-immunological defense of the oral cavity

En este artículo se describen: 1) las características físico-químicas de las mucinas salivales, denominadas MG1 y MG2. 2) El mecanismo de secreción por estimulación simpática y parasimpática. 3) La distinta participación de MG1 y MG2 tanto en la actividad deglutoria como en los mecanismos de defensa de la cavidad bucal, en relación con sus propiedades físico-químicas. 4) El rol de las mucinas salivales en la protección de la mucosa del tracto gastrointestinal. 5) La relación entre las mucinas saliales y las patologías de la cavidad bucal.(AU)

Mucinas salivales: estructura química, mecanismos de liberación y participación en la defensa no inmunológica de la cavidad bucal / Salivary mucins: chemical structure, release mechanisms and participation in the non-immunological defense of the oral cavity

En este artículo se describen: 1) las características físico-químicas de las mucinas salivales, denominadas MG1 y MG2. 2) El mecanismo de secreción por estimulación simpática y parasimpática. 3) La distinta participación de MG1 y MG2 tanto en la actividad deglutoria como en los mecanismos de defensa de la cavidad bucal, en relación con sus propiedades físico-químicas. 4) El rol de las mucinas salivales en la protección de la mucosa del tracto gastrointestinal. 5) La relación entre las mucinas saliales y las patologías de la cavidad bucal.

MUC1 expression and anti-MUC1 serum immune response in head and neck squamous cell carcinoma (HNSCC): a multivariate analysis.

BACKGROUND: HNSCC progression to adjacent tissue and nodes may be mediated by altered glycoproteins and glycolipids such as MUC1 mucin. This report constitutes a detailed statistical study about MUC1 expression and anti-MUC1 immune responses in relation to different clinical and pathological parameters which may be useful to develop new anti HNSCC therapeutic strategies. PATIENTS AND METHODS: Fifty three pre treatment HNSCC patients were included: 26 (49.1%) bearing oral cavity tumors, 17 (32.1%) localized in the larynx and 10 (18.8%) in the pharynx. Three patients (5.7%) were at stage I, 5 (9.4%) stage II, 15 (28.3%) stage III and 30 (56.6%) at stage IV. MUC1 tumor expression was studied by immunohistochemistry employing two anti-MUC1 antibodies: CT33, anti cytoplasmic tail MUC1 polyclonal antibody (Ab) and C595 anti-peptidic core MUC1 monoclonal antibody. Serum levels of MUC1 and free anti-MUC1 antibodies were detected by ELISA and circulating immune complexes (CIC) by precipitation in polyethylene glycol (PEG) 3.5%; MUC1 isolation from circulating immune complexes was performed by protein A-sepharose CL-4B affinity chromatography followed by SDS-PAGE and Western blot. Statistical analysis consisted in Multivariate Principal Component Analysis (PCA); ANOVA test (Tukey's test) was employed to find differences among groups; nonparametrical correlations (Kendall's Tau) were applied when necessary. Statistical significance was set to p < 0.05 in all cases. RESULTS: MUC1 cytoplasmic tail was detected in 40/50 (80%) and MUC1 protein core in 9/50 (18%) samples while serum MUC1 levels were elevated in 8/53 (15%) patients. A significant statistical correlation was found between MUC1 serum levels and anti-MUC1 IgG free antibodies, while a negative correlation between MUC1 serum levels and anti-MUC1 IgM free antibodies was found. Circulating immune complexes were elevated in 16/53 (30%) samples and were also statistically associated with advanced tumor stage. MUC1 was identified as an antigenic component of IgG circulating immune complexes. Moreover, poorly differentiated tumors were inversely correlated with tumor and serum MUC1 detection and positively correlated with node involvement and tumor mass. CONCLUSION: Possibly, tumor cells produce MUC1 mucin which is liberated to the circulation and captured by IgG antibodies forming MUC1-IgG-CIC. Another interesting conclusion is that poorly differentiated tumors are inversely correlated with tumor and serum MUC1 detection.

Immunohistochemical analysis of MUC5B apomucin expression in breast cancer and non-malignant breast tissues.

A deregulation of several MUC genes (MUC1, MUC2, MUC3, MUC5AC, and MUC6) was previously demonstrated in breast carcinomas. Considering that recently we found the "non-mammary" MUC5B mRNA in primary breast tumors (Berois et al. 2003), we undertook the present study to evaluate the expression profile of MUC5B protein product in breast tissues, using LUM5B-2 antisera raised against sequences within the non-glycosylated regions of this apomucin. Expression of MUC5B by breast cancer cells was confirmed by immunocytochemistry, in situ hybridization, and Western blot on MCF-7 cancer cells. Using an immunohistochemical procedure, MUC5B apomucin was detected in 34/42 (81%) primary breast tumors, in 13/14 (92.8%) samples of non-malignant breast diseases, in 8/19 (42.1%) samples of normal-appearing breast epithelia adjacent to cancer, and in 0/5 normal control breast samples. The staining pattern of MUC5B was very different when comparing breast cancer cells (cytoplasmic) and non-malignant breast cells (predominantly apical and in the secretory material). We analyzed MUC5B mRNA expression using RT-PCR in bone marrow aspirates from 22/42 patients with breast cancer to compare with MUC5B protein expression in the primary tumors. Good correlation was observed because the six MUC5B-positive bone marrow samples also displayed MUC5B expression in the tumor. Our results show, for the first time at the protein level, that MUC5B apomucin is upregulated in breast cancer. Its characterization could provide new insights about the glycobiology of breast cancer cells.

Detection of bone marrow-disseminated breast cancer cells using an RT-PCR assay of MUC5B mRNA.

The evaluation of disseminated epithelial tumor cells in breast cancer patients has generated considerable interest due to its potential association with disease recurrence. Our work was performed to analyze the usefulness of 5 mucin genes expression (MUC2, MUC3, MUC5B, MUC6 and MUC7), using RT-PCR assays, to detect disseminated cancer cells in patients with operable breast cancer. The highest frequencies of positive RT-PCR tests in breast tumor extracts were observed for MUC5B (7/15) and MUC7 (5/12). The best specificity, negative results on all peripheral blood mononuclear (PBMN) cell samples from healthy donors, were shown for MUC2, MUC5B and MUC6 RT-PCR assays. Thus, we selected MUC5B as a target gene for further evaluation. Using a nested RT-PCR, MUC5B mRNA transcripts were detected in 16/31 primary breast tumors (but not in 36 samples of normal PBMN cells) and in the human MCF-7 breast cancer cell line but not in BT20, MDA, T47D and ZR-75 breast cancer cell lines, indicating that MUC5B mRNA is expressed in a population of breast cancer cells. Using this method, 9/46 patients (19.5%) who underwent curative surgery showed positive MUC5B mRNA in bone marrow aspirates obtained prior to surgery, including 5/24 patients (20.8%) with stage I or II breast cancer, without histopathologic lymph node involvement. These results indicate that MUC5B mRNA could be a specific marker applicable to the molecular diagnosis of breast cancer cell dissemination. A comparative evaluation between MUC5B mRNA, cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA in all bone marrow aspirates suggests a putative complementation for molecular detection of disseminated carcinoma cells. Considering that breast cancer is characterized by a great phenotypic heterogeneity, the use of multimarker approach could contribute to tumor cell detection in bone marrow and blood.

Electrophoretic analysis of whole saliva and prevalence of dental caries. A study in Mexican dental students.

BACKGROUND: Variability in salivary proteins and their posttranslational modifications may play an important role in determining their protective features against dental caries. Knowledge of molecular content of saliva in different populations is important for a better understanding of protective properties of this biological fluid. Aims of this study were to analyze electrophoretic pattern and protein composition in resting human whole saliva (HWS) of a Mexican population and to correlate these data with decayed, missing, and filled teeth (DMFT) index in these subjects. METHODS: Resting human whole saliva samples were collected from 120 healthy Mexican dental students. Salivary flow rate, protein concentration, and electrophoretic profile analyzed qualitatively by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were correlated with DMFT index. Gels were successively triple-stained with Coomassie brilliant blue R250, periodic acid Schiff (PAS), silver stain, and salivary molecules were scored as absent (-), present (+/-), and high intensity and size (+). RESULTS: These showed no substantial differences in number of bands between males and females; however, a slight correlation between total protein concentration and sex was found (p

Breast cancer associated mucin: a review.

Breast mucins are expressed by malignant epithelial cells and they elicit an immune reaction. The up-regulation of mucin expression is association with tumour invasion, this mucin called MUC-1 reduces the cell-cell interaction facilitating cell detachment. The MUC-1 gene product, known as polymorphic epithelial mucin is a transmembrane high molecular weight glycoprotein. The molecule of MUC-1 has a central polypeptidic core with a carbohydrate linked in O-linkage to serines and threonines. The carbohydrate side chain epitope of MUC-1 molecule produced by breast cancer cells is heavily sialylated, giving their physical properties and increasing their immunogenicity. The development of monoclonal antibodies (MAb) has led to study the MUC-1 in subcellular extracts, tissues and culture supernatants from breast cancer and also colorectal carcinoma. The pattern of tumour cell staining with labeled MAb varies according with the grade of malignancy; these MAb bind either to peptide sequence and/or to the glycosylated epitopes. MUC-1 has a clinical relevance because serum concentrations may be useful for monitoring the response to therapy and progress of disease. MUC-1 epitope masking has been described since specific antibodies can combine with them forming immune complexes. Finally, mucins have been considered to develop vaccines against cancer, targeting specific carbohydrate and mucin epitopes.

Rapid expulsion of the intestinal trematodes Echinostoma trivolvis and E. caproni from C3H/HeN mice after infection with Nippostrongylus brasiliensis.

The number of goblet cells in the small intestines of C3H/HeN mice increased rapidly following their infection with about 500 third-stage larvae (L3) of the intestinal nematode Nippostrongylus brasiliensis. The number of goblet cells reached its peak on day 9 postinfection (p.i.). Worm burdens in the hosts' small intestines were determined following a challenge infection with encysted metacercariae of the intestinal trematodes Echinostoma trivolvis or E. caproni on days 8 and 16 after primary infections with N. brasiliensis. All metacercariae of E. trivolvis or E. caproni used to challenge the hosts on day 8 p.i. were expelled. Considerable numbers of E. trivolvis (48.6%) and E. caproni (67.1%) remained in the intestines of hosts challenged with these echinostomes on day 16 p.i. All the controls used for E. trivolvis and E. caproni infections without primary infections with N. brasiliensis showed recovery rates greater than 70%. An enzyme-linked immunosorbent assay (ELISA) showed that the IgM titer rose remarkably and plateaued on day 11 p.i. No marked rise in the IgG or IgA titer occurred during the experiment. These results indicate that mucins increased by hyperplastic goblet cells associated with primary infections with N. brasiliensis are responsible for a rapid expulsion of the worms of the challenge infection with E. trivolvis or E. caproni from the mouse host.

The protozoan Trypanosoma cruzi has a family of genes resembling the mucin genes of mammalian cells.

Mucins are heavily O-glycosylated Thr/Ser/Pro-rich molecules. Given their relevant functions, mucins and their genes have been mainly studied in higher eukaryotes. In the protozoan parasite Trypanosoma cruzi, mucin-like glycoproteins were shown to play an important role in the interaction with the surface of the mammalian cell during the invasion process. We show now that this parasite has a family of putative mucin genes, whose organization resembles the one present in mammalian cells. Different parasite isolates have different sets of genes, as defined by their central domain. Central domains, rich in codons for Thr and/or Ser and Pro residues, are made up of either a variable number of repeat units in tandem or non-repetitive sequences. Conversely, 5'- and 3'-ends from different genes in different isolates have similar sequences, suggesting their common origin. Comparison of deduced amino acid sequences revealed that all members of the family have the same putative signal peptide on the N terminus and a putative sequence for glycophosphatidylinositol anchoring on the C terminus. The deduced molecular mass of the core proteins is small (from 17 to 21 kDa), in agreement with the 1-kilobase size of the mRNA detected. Putative mucin genes in T. cruzi are located on large chromosomal bands of about 1.6-2.2 megabase pairs.

Embryogeny of human labial glands: a structural, ultrastructural and cytochemical study.

A histochemical study of labial glands was performed to compare the different stages of differentiation with those of lingual glands previously studied. Labial glands of 8 to 32 week old human fetuses were analyzed with Hematoxylin/eosine, PAS, Cason, Alcian blue, Toluidine blue, methenamine/silver, TEM and Ruthenium red techniques. At 8-10 weeks various differentiation phases of cell cords originated in the epithelium of the labial mucosa were observed. Acinar buds had PAS positive, alcianophilic and metachromatic material in the lumen of 14 week labial glands. The excretory ducts featured similar characteristics. At 24 weeks groups of mucous and seromucous acini were identified and the mucosubstances increased in the 32 week old fetuses. These results show that the labial glands are histophysiologically differentiated at an earlier stage of development (14 weeks) as compared to lingual glands (20 weeks). However, mucosubstance production would begin during the early phases of embryogenesis for both labial and lingual glands.

Histologia, histopatologia e histoquímica de mucinas do fundo e do corpo do estômago em Philander opossum / Histology, histopathology and mucins histochemistry of the stomach fundus and body in Philander opossum

Foram realizados estudos histológicos, histopatológicos e histoquímicos de mucinas do epitélio gástrico do fundo e do corpo de seis cuicas de quatro olhos (PPhilander opossum), adultos e de ambos os sexos. Os animais forma separados em dois grupos e submetidos a dois tipos diferentes de alimentaçäo. A morfologia do fundo e do corpo gástrico da cuica segue o padräo histológico geral para a maioria dos mamíferos. Nos animais submetidos a alimentaçäo natural, a mucosa apresentou-se com características normais. Naqueles cujos hábitos alimentares näo foram preservados após a captura e mantidos em biotério, verificou-se a ocorrência de células com significativa degeneraçäo hidrópica no epitélio de revestimento da mucosa, das fossetas e nas glândulas. Esta degeneraçäo foi mais intensa nas células parietais. Sugere-se ao se trabalhar com animais silvestres, sacrificá-los logo após a captura ou proporcionar aos mesmos alimentaçäo adequada. A mudança de alimentaçäo pode causar lesöes no estômago. Mucina neutras foram evidenciada no epitélio de revestimento da mcuosa, das fossetas e nas células mucosas do colo de todos os animais estudados

Aspectos histológicos, histopatológicose histoquímicos de mucinas do epitélio gástrico da regiäo pilórica de Philander opossum / Some histological, histopathological and histochemical (Mucins) aspects of the gastric epitheluim of pyloric region in Phelander Opossum

Estudos histológicos, histopatológicos e histoquímicos de mucinas do epitélio gástrico da regiäo pilórica de seis cuicas em jejum, foram realizados. Os animais eram adultos, de ambos os sexos e foram separados em dois grupos submetidos a diferentes dietas alimentares. A morfologia da regiäo pilórica da cuica (Philander opossum) segue o plano histológico geral para a maioria dos mamíferos. Nos animais submetidos `a alimentaçäo natural e mucosa apresentou-se normal. Naqueles cujos hábitos alimentares näo foram preservados após a captura e mantidos em biotério, verificou-se a ocorrência de células com significativa degeneraçäo hidrópica no epitélio de revestimento da mucosa e nas fovéolas. Nas células secretoras das glândulas pilóricas a degeneraçäo hidrópica foi discreta. Sugere-se ao se trabalhar com animais silvestres, sacrificá-los logo após a captura ou maior cuidado na preservaçäo de seus hábitos alimentares. A mudança de alimentaçäo pode causar lesöes no estômago. Mucinas neutras foram detectadas no epiélio de revestimento da mucosa, epitélio das fovéolas e nas glândulas pilóricas. Näo houve diferenças nas mucinas detectadas nos animais submetidos `as diferentes condiçöes de alimentaçäo