The Acute Inflammatory Potential of Particles From the Echinococcus granulosus Laminated Layer Is Moderated by Its Calcium Inositol Hexakisphosphate Component.

Cystic echinococcosis is caused by the tissue-dwelling larva (hydatid) of Echinococcus granulosus sensu lato. A salient feature is that this larva is protected by the acellular laminated layer (LL). As the parasite grows, the LL sheds abundant particles that can accumulate in the parasite's vicinity. The potential of LL particles to induce inflammation in vivo has not been specifically analysed. It is not known how each of its two major components, namely highly glycosylated mucins and calcium inositol hexakisphosphate (InsP6) deposits, impacts inflammation induced by the LL as a whole. In this work, we show that LL particles injected intraperitoneally cause infiltration of eosinophils, neutrophils and monocytes/macrophages as well as the disappearance of resident (large peritoneal) macrophages. Strikingly, the absence of calcium InsP6 enhanced the recruitment of all the inflammatory cell types analysed. In contrast, oxidation of the mucin carbohydrates caused decreased recruitment of neutrophils. The carbohydrate-oxidised particles caused cell influx nonetheless, which may be explained by possible receptor-independent effects of LL particles on innate immune cells, as suggested by previous works from our group. In summary, LL particles can induce acute inflammatory cell recruitment partly dependent on its mucin glycans, and this recruitment is attenuated by the calcium InsP6 component.

Role of the Fungus Pneumocystis in IL1ß Pathway Activation and Airways Collagen Deposition in Elastase-Induced COPD Animals.

Inflammation and mucus production are prevalent characteristics of chronic respiratory conditions, such as asthma and chronic chronic obstructive pulmonary disease (COPD). Biological co-factors, including bacteria, viruses, and fungi, may exacerbate these diseases by activating various pathways associated with airway diseases. An example is the fungus Pneumocystis, which is linked to severe COPD in human patients. Recent evidence has demonstrated that Pneumocystis significantly enhanced inflammation and mucus hypersecretion in a rat model of elastase-induced COPD. The present study specifically aims to investigate two additional aspects associated with the pathology induced by Pneumocystis infection: inflammation and collagen deposition around airways. To this end, the focus was to investigate the role of the IL-1ß pro-inflammatory pathway during Pneumocystis infection in COPD rats. Several airway pathology-related features, such as inflammation, mucus hypersecretion, and fibrosis, were evaluated using histological and molecular techniques. COPD animals infected with Pneumocystis exhibited elevated inflammation levels, including a synergistic increase in IL-1ß and Cox-2. Furthermore, protein levels of the IL-1ß-dependent transcription factor cAMP response element-binding (CREB) showed a synergistic elevation of their phosphorylated version in the lungs of COPD animals infected with Pneumocystis, while mucus levels were notably higher in the airways of COPD-infected animals. Interestingly, a CREB responsive element (CRE) was identified in the Muc5b promoter. The presence of CREB in the Muc5b promoter was synergistically increased in COPD animals infected with Pneumocystis compared to other experimental groups. Finally, an increment of deposited collagen was identified surrounding the airways of COPD animals infected with Pneumocystis compared with the other experimental animal groups and correlated with the increase of Tgfß1 mRNA levels. These findings emphasize the role of Pneumocystis as a potential biological co-factor in chronic respiratory diseases like COPD or asthma, warranting new perspectives in the treatment of chronic respiratory diseases.

Histochemistry and transcriptomics of mucins and peritrophic membrane (PM) proteins along the midgut of a beetle with incomplete PM and their complementary function.

The midgut of Zabrotes subfasciatus (Coleoptera) and other insects may have regions lacking a peritrophic membrane (matrix, PM) and covered with a jelly-like material known as peritrophic gel. This work was undertaken to test the hypothesis that the peritrophic gel is a vertebrate-like mucus. By histochemistry we identified mucins along the whole midgut, which contrasts with the known occurrence of PM only at the posterior midgut. We also analyzed the expression of the genes coding for mucus-forming mucins (Mf-mucins), peritrophins, chitin synthases and chitin deacetylases along the midgut and carcass (insect without midgut) by RNA-seq. Mf-mucins were identified as proteins with high O-glycosylation and multiple tandem repeats of Pro/Thr/Ser residues. Peritrophins were separated into PM proteins, cuticular proteins analogous to peritrophins (CPAPs) and ubiquitous-chitin-binding domain-(CBD)-containing proteins (UCBPs). PM proteins have at least 3, CPAP one or 3, and UCBPs have a varied number of CBDs. PM proteins are more expressed at midgut, CPAP at the carcass, and UCBP at both. The results showed that most PM proteins are mainly expressed at the posterior midgut, together with midgut chitin synthase and chitin deacetylase, and in agreement with the presence of PM only at the posterior midgut by visual inspection. The excretion of most midgut chitinase is avoided, suggesting that the shortened PM is functional. Mf-mucins are expressed along the whole midgut, probably forming the extracellular mucus layer observed by histochemistry. Thus, the lack of PM at anterior and middle midgut causes the exposure of a mucus, which may correspond to the previously described peritrophic gel. The putative functional interplay of mucus and PM is discussed. The major role of mucus is proposed to be tissue protection and of PM to enhancing digestive efficiency by allowing enzyme recycling.

Pneumocystis Exacerbates Inflammation and Mucus Hypersecretion in a Murine, Elastase-Induced-COPD Model.

Inflammation and mucus hypersecretion are frequent pathology features of chronic respiratory diseases such as asthma and COPD. Selected bacteria, viruses and fungi may synergize as co-factors in aggravating disease by activating pathways that are able to induce airway pathology. Pneumocystis infection induces inflammation and mucus hypersecretion in immune competent and compromised humans and animals. This fungus is a frequent colonizer in patients with COPD. Therefore, it becomes essential to identify whether it has a role in aggravating COPD severity. This work used an elastase-induced COPD model to evaluate the role of Pneumocystis in the exacerbation of pathology, including COPD-like lung lesions, inflammation and mucus hypersecretion. Animals infected with Pneumocystis developed increased histology features of COPD, inflammatory cuffs around airways and lung vasculature plus mucus hypersecretion. Pneumocystis induced a synergic increment in levels of inflammation markers (Cxcl2, IL6, IL8 and IL10) and mucins (Muc5ac/Muc5b). Levels of STAT6-dependent transcription factors Gata3, FoxA3 and Spdef were also synergically increased in Pneumocystis infected animals and elastase-induced COPD, while the levels of the mucous cell-hyperplasia transcription factor FoxA2 were decreased compared to the other groups. Results document that Pneumocystis is a co-factor for disease severity in this elastase-induced-COPD model and highlight the relevance of STAT6 pathway in Pneumocystis pathogenesis.

Mucin levels in glands of the colonic mucosa of rats with diversion colitis subjected to enemas containing sucralfate and n-acetylcysteine alone or in combination

Purpose: To evaluate the tissue content of neutral and acidic mucins, sulfomucins and sialomucins in colonic glands devoid of intestinal transit after enemas containing sucralfate and n-acetylcysteine alone or in combination. Methods: Sixty-four rats underwent intestinal transit bypass. A colonic segment was collected to compose the white group (without intervention). After derivation, the animals were divided into two groups according to whether enemas were performed daily for two or four weeks. Each group was subdivided into four subgroups according to the substance used: control group: saline 0.9%; sucralfate group (SCF): SCF 2 g/kg/day; n-acetylcysteine group (NAC): NAC 100 mg/kg/day; and SCF+NAC group: SCF 2 g/kg/day + NAC 100 mg/kg/day.Neutral and acidic mucins were stained by periodic acid-Schiff and alcian-blue techniques, respectively. The distinction between sulfomucins and sialomucin was made by the high alcian-blue iron diamine technique. The content of mucins in the colonic glands was measured by computerized morphometry. The inflammatory score was assessed using a validated scale. The results between the groups were compared by the Mann-Whitney's test, while the variation according to time by the Kruskal-Wallis' test (Dunn's post-test). A significance level of 5% was adopted. Results: There was reduction in the inflammatory score regardless of the application of isolated or associated substances. Intervention with SCF+NAC increased the content of all mucin subtypes regardless of intervention time. Conclusions: The application of SCF+NAC reduced the inflammatory process of the colonic mucosa and increased the content of different types of mucins in the colonic glands of segments excluded from fecal transit.

Liquen mixedematoso atípico asociado a mieloma múltiple / Atypical lichen myxedematosus associated to multiple myeloma

Resumen El liquen mixedematoso (LM) representa un grupo de enfermedades cutáneas raras, el cual se encuentra dentro de las mucinosis crónicas. Anteriormente descrita como escleromixedema localizado, sin embargo, a diferencia de éste, por lo general no tiene compromiso sistémico. Dentro de los subtipos, se encuentra el LM atípico, el cual es infrecuente y hay pocos casos reportados asociados a mieloma múltiple (MM). Se presenta el caso de un paciente masculino con MM positivo para cadenas lambda, con cuadro clínico de inicio agudo, en quien se realizó diagnóstico de LM atípico; recibió manejo con corticoide tópico con mejoría de las lesiones al mes de tratamiento.

Abstract Lichen myxedematous (LM) represents a group of rare skin diseases, which is found within the chronic mucinoses. Previously described as localized scleromyxedema, however, unlike localized scleromyxedema, it usually does not have systemic involvement. Among the subtypes, there is atypical LM, which is infrequent and there are few reported cases associated with multiple myeloma (MM). We present the case of a male patient with MM positive for lambda chains, with acute onset clinical picture, who was diagnosed with atypical LM; he received management with topical corticosteroid with improvement of the lesions after one month of treatment.

Accessing the Variability of Multicopy Genes in Complex Genomes using Unassembled Next-Generation Sequencing Reads: The Case of Trypanosoma cruzi Multigene Families.

Repetitive elements cause assembly fragmentation in complex eukaryotic genomes, limiting the study of their variability. The genome of Trypanosoma cruzi, the parasite that causes Chagas disease, has a high repetitive content, including multigene families. Although many T. cruzi multigene families encode surface proteins that play pivotal roles in host-parasite interactions, their variability is currently underestimated, as their high repetitive content results in collapsed gene variants. To estimate sequence variability and copy number variation of multigene families, we developed a read-based approach that is independent of gene-specific read mapping and de novo assembly. This methodology was used to estimate the copy number and variability of MASP, TcMUC, and Trans-Sialidase (TS), the three largest T. cruzi multigene families, in 36 strains, including members of all six parasite discrete typing units (DTUs). We found that these three families present a specific pattern of variability and copy number among the distinct parasite DTUs. Inter-DTU hybrid strains presented a higher variability of these families, suggesting that maintaining a larger content of their members could be advantageous. In addition, in a chronic murine model and chronic Chagasic human patients, the immune response was focused on TS antigens, suggesting that targeting TS conserved sequences could be a potential avenue to improve diagnosis and vaccine design against Chagas disease. Finally, the proposed approach can be applied to study multicopy genes in any organism, opening new avenues to access sequence variability in complex genomes. IMPORTANCE Sequences that have several copies in a genome, such as multicopy-gene families, mobile elements, and microsatellites, are among the most challenging genomic segments to study. They are frequently underestimated in genome assemblies, hampering the correct assessment of these important players in genome evolution and adaptation. Here, we developed a new methodology to estimate variability and copy numbers of repetitive genomic regions and employed it to characterize the T. cruzi multigene families MASP, TcMUC, and transsialidase (TS), which are important virulence factors in this parasite. We showed that multigene families vary in sequence and content among the parasite's lineages, whereas hybrid strains have a higher sequence variability that could be advantageous to the parasite's survivability. By identifying conserved sequences within multigene families, we showed that the mammalian host immune response toward these multigene families is usually focused on the TS multigene family. These TS conserved and immunogenic peptides can be explored in future works as diagnostic targets or vaccine candidates for Chagas disease. Finally, this methodology can be easily applied to any organism of interest, which will aid in our understanding of complex genomic regions.

The α-Galactosyl Carbohydrate Epitope in Pathogenic Protozoa.

The α-gal epitope, which refers to the carbohydrate α-d-Galp-(1 → 3)-ß-d-Galp-(1 → 4)-d-GlcNAc-R, was first described in the glycoconjugates of mammals other than humans. Evolution caused a mutation that resulted in the inactivation of the α-1,3-galactosyltransferase gene. For that reason, humans produce antibodies against α-d-Galp containing glycoproteins and glycolipids of other species. We summarize here the glycoconjugates with α-d-Galp structures in Trypanosoma, Leishmania, and Plasmodium pathogenic protozoa. These were identified in infective stages of Trypanosoma cruzi and in Plasmodium sporozoites. In Leishmania, α-d-Galp is linked differently in the glycans of glycoinositolphospholipids (GIPLs). Chemically synthesized neoglycoconjugates have been proposed as diagnostic tools and as antigens for vaccines. Several syntheses reported for the α-gal trisaccharide, also called the Galili epitope, and the glycans of GIPLs found in Leishmania, the preparation of neoglycoconjugates, and the studies in which they were involved are also included in this Review.

New algorithm to differentiate histochemical types of intestinal metaplasia: G&S2 method / Novo algoritmo para diferenciar tipos histoquímicos de metaplasia intestinal: G & Amp; Método S2

ABSTRACT Introduction The classification of intestinal metaplasia by histochemistry methods has been described as the most appropriate. However, current algorithms are not replicable in laboratories due to severe limitations. Objective To evaluate a new algorithm to differentiate histochemical types of intestinal metaplasia. Material and Method: Cross-sectional research in which 512 gastric biopsies with intestinal metaplasia (paraffin blocks) were evaluated by a new algorithm using two types of Alcian Blue dyes during February-March of 2020 in the pathological anatomy service of the Maria Auxiliadora Hospital, Lima, Peru. This evaluation consisted of two steps: visualization of acid mucins in the columnar cells of the gastric mucosa and calculation of the weighted Kappa statistic. Results Histochemical types of intestinal metaplasia showed as follows: Type I, 398 (77.7%); Type II, 81 (15.8%) and Type III, 33 (6.5%). The weighted Kappa statistic was 0.79 (p<0.001), rated as an important or good concordance. Conclusion This new algorithm demonstrated it was useful and capable of identifying and differentiating the histochemical types of intestinal metaplasia, in addition to having statistical reliability.

RESUMO Introdução A classificação da metaplasia intestinal por métodos histoquímicos tem sido descrita como a mais adequada. No entanto, os algoritmos atuais não são replicáveis?? em laboratórios devido a limitações severas. Objetivo Avaliar um novo algoritmo para diferenciar tipos histoquímicos de metaplasia intestinal. Material e Método: Pesquisa transversal em que 512 biópsias gástricas com metaplasia intestinal (blocos de parafina) foram avaliadas por um novo algoritmo usando dois tipos de corantes Alcian Blue durante fevereiro-março de 2020 no serviço de anatomia patológica do Hospital Maria Auxiliadora, Lima, Peru. Essa avaliação consistiu em duas etapas: visualização das mucinas ácidas nas células colunares da mucosa gástrica e cálculo da estatística Kappa ponderado. Resultados Os tipos histoquímicos de metaplasia intestinal mostraram-se a seguir: Tipo I, 398 (77,7%); Tipo II, 81 (15,8%) e Tipo III, 33 (6,5%). A estatística Kappa ponderado foi de 0,79 (p <0,001), classificada como concordância importante ou boa. Conclusão Este novo algoritmo demonstrou ser útil e capaz de identificar e diferenciar os tipos histoquímicos de metaplasia intestinal, além de possuir confiabilidade estatística

Histochemical assessment of mucin-secreting cells in the stomach of domestic rabbit / Evaluación histoquímica de células secretoras de mucina en el estómago de conejo doméstico

SUMMARY: The mucous substances of the stomach in mammals are important not only for the protection of the gastric epithelium from the acid environment and grinding actions, but it facilitates some other functions of the stomach such as antibacterial, antimetastatic, and immunological roles. The goal of the study is to highlight the distribution of mucin-secreting cells in the gastric mucosa in domestic rabbits, including the type of mucus synthesized. The gastric samples collected from ten individual rabbits were fixed in 10 % buffered formalin and underwent later standard paraffin tissue sample processing, which included dehydration, clarification, and embedding in paraffin. The tissue sections were eventually stained histochemically by PAS reaction and by Alcian blue method (pH 2.5) for neutral and acidic mucins detection, respectively. The quantification of mucins in the cytoplasm of mucus-secreting cells was performed by grading the gastric tissue samples from negative (-) to intensely positive (++). The mucus elaboration was observed in all the regions of the stomach (i.e., cardial, fundic, and pyloric regions), but only for the neutral mucin. The acidic mucin synthesis occurred only in the secretory units of the gastric glands from the cardial region in the stomach. Pyloric glands synthesized the largest amounts of neutral mucins, followed by moderate amounts elaborated by cardial glands, while the fundic region does not synthesize it at all. The description of new microscopic features of the stomach in rabbits is fundamental not only for comprehending species-related physiological features but gastric pathological processes.

RESUMEN: Las sustancias mucosas del estómago en los mamíferos son importantes no solo para la protección del epitelio gástrico del ambiente ácido y las acciones de trituración, sino que facilitan además otras funciones del estómago, como son las funciones antibacterianas, antimetastásicas e inmunológicas. El objetivo del estudio fue resaltar la distribución de las células secretoras de mucina en la mucosa gástrica de conejos domésticos, incluido el tipo de moco sintetizado. Las muestras gástricas recolectadas de diez conejos se fijaron en formalina tamponada al 10 % y se sometieron a un procesamiento que incluyó deshidratación, clarificación e inclusión en parafina. Las secciones de tejido finalmente se tiñeron histoquímicamente mediante la reacción de PAS y el método del azul de Alcian (pH 2,5) para la detección de mucinas neutras y ácidas, respectivamente. La cuantificación de mucinas en el citoplasma de las células secretoras de moco se realizó clasificando las muestras de tejido gástrico desde negativas (-) hasta intensamente positivas (++). La elaboración de moco se observó en todas las regiones del estómago (es decir, cardias, fúndica y pilórica), pero solo para la mucina neutra. La síntesis de mucina ácida ocurrió solo en las unidades secretoras de las glándulas gástricas de la región correspondiente al cardias del estómago. Las glándulas pilóricas sintetizaron la mayor cantidad de mucinas neutras, seguidas de cantidades moderadas elaboradas por las glándulas cardiales, mientras que la región fúndica no las sintetizó en abso- luto. La descripción de nuevas características microscópicas del estómago en conejos es fundamental no solo para comprender las características fisiológicas relacionadas con las especies sino también para entender los procesos patológicos gástricos.

Characterization of Oral Immunity in Cases and Close Household Contacts Exposed to Andes Orthohantavirus (ANDV).

Background: Andes orthohantavirus (ANDV) is the sole etiologic agent of Hantavirus Cardiopulmonary Syndrome in Chile and, until now, the only Hantavirus known to be transmitted by person-to-person route. The main risk of person-to-person transmission is to be a sexual partner of an index case, and deep kissing the main mechanism of infection. Experimental reports suggest that ANDV infection can be inhibited by some saliva components. Therefore, some host factors like saliva quality, could help to explain why some individuals do not become infected even though their exposure to the virus is high. Aim: To compare some saliva components, such cytokines and mucins, between ANDV-infected cases (exposed-sick), their close household contacts (exposed-not sick) and healthy control not exposed. Methods: Sixty-nine confirmed ANDV-infected cases, 76 close household contacts exposed to ANDV but not infected (CHC) and 39 healthy control not exposed (HCNE). The following components were measured in saliva: secretory immunoglobulin A (sIgA) by ELISA; cytokines (IL1ß, IL12p70, TNFα, INFy, IL10, IL6, VEGF, IP10, and IL8) by Multiplex Assay and mucins MUC7 and MUC5B by Western Blotting. Results: Among infected cases, CHC and HCNE analyzed 74, 45, and 33% were men, respectively (p ≤ 0.05). The average age for cases, CHC and HCNE was 37.7, 28.7, and 32 years, respectively (p ≤ 0.05). The average concentration of sIgA in infected cases was 4.846 mg/mL, higher than for CHC group, 0.333 mg/mL (p ≤ 0.05). For cytokines, significant differences were found comparing all groups for IFNy, IL12p70, and IL8. Among CHC group, there was a higher frequency of detection of MUC7 isoform (62.6%; 31/49) compared to ANDV-infected cases (40.5%; 17/42) (p < 0.05). Similarly, presence of MUC5B was higher among CHC groups (62.16%; 46/74) than in ANDV-infected cases (44.4%; 28/63) (p < 0.05). Conclusions: Three salivary components showed differences between infected cases and close household contacts (sIgA, cytokines, and mucins). These differences can be explained by the acute state of the disease in the ANDV-infected cases group. However, the differences in MUC5B and isoforms of MUC7 are not entirely explainable by the infection itself. This work represents a novel description of salivary components in the context of ANDV infection.

The Glycan Structure of T. cruzi mucins Depends on the Host. Insights on the Chameleonic Galactose.

Trypanosoma cruzi, the protozoa that causes Chagas disease in humans, is transmitted by insects from the Reduviidae family. The parasite has developed the ability to change the structure of the surface molecules, depending on the host. Among them, the mucins are the most abundant glycoproteins. Structural studies have focused on the epimastigotes and metacyclic trypomastigotes that colonize the insect, and on the mammal trypomastigotes. The carbohydrate in the mucins fulfills crucial functions, the most important of which being the accepting of sialic acid from the host, a process catalyzed by the unique parasite trans-sialidase. The sialylation of the parasite influences the immune response on infection. The O-linked sugars have characteristics that differentiate them from human mucins. One of them is the linkage to the polypeptide chain by the hexosamine, GlcNAc, instead of GalNAc. The main monosaccharide in the mucins oligosaccharides is galactose, and this may be present in three configurations. Whereas ß-d-galactopyranose (ß-Galp) was found in the insect and the human stages of Trypanosoma cruzi, ß-d-galactofuranose (ß-Galf) is present only in the mucins of some strains of epimastigotes and α-d-galactopyranose (α-Galp) characterizes the mucins of the bloodstream trypomastigotes. The two last configurations confer high antigenic properties. In this review we discuss the different structures found and we pose the questions that still need investigation on the exchange of the configurations of galactose.

Biomarcadores para la evaluación y diagnóstico del síndrome de ojo seco: una revisión / Biomarkers for the evaluation and diagnosis of the dry eye syndrome: a review

Resumen Introducción: El síndrome de ojo seco es una enfermedad en la que se generan signos y síntomas que conducen a alteraciones oculares prolongadas, por lo tanto, es relevante establecer con precisión la etiología de la enfermedad con la finalidad de establecer el tratamiento más efectivo, de allí, la importancia del desarrollo de exámenes innovadores como son los biomarcadores, los cuales permiten identificar con mayor precisión el cuadro clínico. Por esta razón, el presente trabajo pretende describir los principales avances de los biomarcadores de la superficie ocular y reconocer su aplicación clínica para el diagnóstico de ojo seco entre los años 2013 a 2018. Metodología: Se analizó literatura sobre biomarcadores empleados para el diagnóstico del ojo seco, mediante una revisión sistemática tipo narrativa de 2013 a 2018 por medio de los descriptores controlados "Dry Eye Syndrome" "biomarkers" "tear proteins" "eye proteins" seleccionados en DeCS y Pubmed; la búsqueda arrojó 48 estudios, de los cuales seleccionamos 21 para el análisis. Resultados: Son diversas las proteínas lagrimales que pueden ser relacionadas con la presencia y ausencia de la enfermedad, es vital que los biomarcadores sean valorados como una herramienta alternativa para diagnosticar con facilidad y precisión la enfermedad del ojo seco. Discusión: Los biomarcadores permiten reconocer los procesos patógenos y biológicos del síndrome de ojo seco, al reflejar el estado de la superficie ocular en presencia o ausencia de signos y síntomas, facilitando el diagnóstico precoz, seguimiento, tratamiento y control de la enfermedad.

Abstract Introduction: Dry eye syndrome is a disease in which signs and symptoms that lead to prolonged ocular alterations occur, therefore, it is relevant to accurately establish the etiology of the disease with the configuration of establishing the most effective treatment, hence the development of innovative exams such as biomarkers selected with greater precision the clinical picture. For this reason, the present work aims to describe the main advances of biomarkers of the ocular surface and to recognize their clinical application for the diagnosis of dry eye between 2013 and 2018. Metodology: Literature on biomarkers used for the diagnosis of dry eye was analyzed, by means of a systematic narrative review from 2013 to 2018 by means of the controlled descriptors "Dry Eye Syndrome" "biomarkers" "tear proteins" "eye proteins" selected in DeCS and Pubmed; The search yielded 48 studies and 21 studies were selected for the analysis. Results: There are several tear proteins that can be related to the presence and absence of the disease, it is vital that biomarkers are evaluated as an alternative tool to easily and accurately diagnose dry eye disease. Discussion: Biomarkers allow to recognize the pathogenic and biological processes of dry eye syndrome, reflecting the state of the ocular surface in the presence or absence of signs and symptoms, facilitating early diagnosis, monitoring, treatment and control of the disease.

Multifaceted antibodies development against synthetic α-dystroglycan mucin glycopeptide as promising tools for dystroglycanopathies diagnostic.

Dystroglycanopathies are diseases characterized by progressive muscular degeneration and impairment of patient's quality of life. They are associated with altered glycosylation of the dystrophin-glycoprotein (DGC) complex components, such as α-dystroglycan (α-DG), fundamental in the structural and functional stability of the muscle fiber. The diagnosis of dystroglycanopathies is currently based on the observation of clinical manifestations, muscle biopsies and enzymatic measures, and the available monoclonal antibodies are not specific for the dystrophic hypoglycosylated muscle condition. Thus, modified α-DG mucins have been considered potential targets for the development of new diagnostic strategies toward these diseases. In this context, this work describes the synthesis of the hypoglycosylated α-DG mimetic glycopeptide NHAc-Gly-Pro-Thr-Val-Thr[αMan]-Ile-Arg-Gly-BSA (1) as a potential tool for the development of novel antibodies applicable to dystroglycanopathies diagnosis. Glycopeptide 1 was used for the development of polyclonal antibodies and recombinant monoclonal antibodies by Phage Display technology. Accordingly, polyclonal antibodies were reactive to glycopeptide 1, which enables the application of anti-glycopeptide 1 antibodies in immune reactive assays targeting hypoglycosylated α-DG. Regarding monoclonal antibodies, for the first time variable heavy (VH) and variable light (VL) immunoglobulin domains were selected by Phage Display, identified by NGS and described by in silico analysis. The best-characterized VH and VL domains were cloned, expressed in E. coli Shuffle T7 cells, and used to construct a single chain fragment variable that recognized the Glycopeptide 1 (GpαDG1 scFv). Molecular modelling of glycopeptide 1 and GpαDG1 scFv suggested that their interaction occurs through hydrogen bonds and hydrophobic contacts involving amino acids from scFv (I51, Y33, S229, Y235, and P233) and R8 and α-mannose from Glycopeptide 1.

Is the colon mucosa affected by ten days of gastric restriction in an animal model?

Purpose: To identify whether the colon mucosa is affected by ten days of gastric restriction in an animal model. Methods: An experimental model of gastric restriction was devised using rats. The animals were submitted to surgical gastrostomy, and a cylindrical loofah was inserted into the stomach. We studied 30 adult male Wistar rats divided into three groups: the stomach restriction group (R10); the sham group (S10), which underwent the same procedure except for the loofah insertion; and the control group (C10). The expression of neutral and acid mucins was evaluated using histochemical techniques. Goblet cells and protein content were compared between groups using generalized estimation equations (GEEs). Bonferronis multiple comparison was applied to identify differences between the groups. All tests considered a 5% significance level. Results: There was an increased expression of neutral mucins, acid mucins and goblet cells in the R10 group. Collagen was also enhanced in the R10 group. Conclusion: The colon mucosa is affected by ten days of gastric restriction in an animal model, increasing neutral mucins, acid mucins and collagen content with trophic maintenance.(AU)

Lichen myxedematosus: a rare group of cutaneous mucinosis

Abstract: Cutaneous mucinoses are a heterogeneous group of dermatoses in which excess deposition of mucin in the dermis gives the skin a waxy appearance, with papules and plaques that can vary from self-healing mucinosis to even disrupting the normal shape of a patient's face, conferring a leonine facies, or be part of life threatening diseases like scleromyxedema. This review will describe the most recent classification on lichen myxedematosus in the generalized (scleromyxedema) and the localized forms, as well as the different organ systems involved in scleromyxedema, diagnostic workup, current management, and prognosis.

Synthesis of the hexasaccharide from Trypanosoma cruzi mucins with the Galp(1 → 2)Galf unit constructed with a superarmed thiogalactopyranosyl donor.

Hexasaccharide ß-D-Galp-(1→ 2)-[ß-D-Galp-(1 → 3)]-ß-D-Galp-(1 → 6)-[ß-D-Galp-(1 → 2)-ß-D-Galf-(1 → 4)]-D-GlcNAc (1) was found O-linked in mucins of Trypanosoma cruzi epimastigotes and metacyclic trypomatigotes. Studies on the biological pathways and functionalities of the mucin oligosaccharides are prompted in order to understand the interactions of these molecules with the insect host. Trisaccharide constituent ß-D-Galp-(1 → 2)-ß-D-Galf-(1 → 4)-D-GlcNAc was constructed from the reducing to the non-reducing end. We discuss the difficulties to introduce a Galp unit at the O-2 position of a partially protected galactofuranosyl unit which were overcome using an anchimerically superarmed donor. By this route and employing a [3 + 3] nitrilium convergent approach hexasaccharide 1 was synthesized in moderate yield.

Enemas with mesalazine increase the tissue contents of mucins in the colonic mucosa devoid of fecal stream

Purpose: To evaluate the inflammatory reaction and measure the content of mucins, in the colonic mucosa without fecal stream submit to intervention with mesalazine. Methods: Twenty-four rats were submitted to a left colostomy and a distal mucous fistula and divided into two groups according to euthanasia to be performed two or four weeks. Each group was divided into two subgroups according daily application of enemas containing saline or mesalazine at 1.0 g/kg/day. Colitis was diagnosed by histological analysis and the inflammatory reaction by validated score. Acidic mucins and neutral mucins were determined with the alcian-blue and periodic acid of Schiff techniques, respectively. Sulfomucin and sialomucin were identified by high iron diamine-alcian blue technique. The tissue contents of mucins were quantified by computer-assisted image analysis. Mann-Whitney test was used to analyze the results establishing the level of significance of 5%. Results: Enemas with mesalazine in colonic segments without fecal stream decreased the inflammation score and increased the tissue content of all subtypes of mucins. The increase of tissue content of neutral, acid and sulfomucin was related to the time of intervention. Conclusion: Mesalazine enemas reduce the inflammatory process and preserve the content of mucins in colonic mucosa devoid of fecal stream.(AU)

Metabolic Labeling of Surface Neo-sialylglyconjugates Catalyzed by Trypanosoma cruzi trans-Sialidase.

Trypanosoma cruzi, the protozoan agent of Chagas disease, has evolved an innovative metabolic pathway by which protective sialic acid (SA) residues are scavenged from host sialylglycoconjugates and transferred onto parasite surface mucin-like molecules (or surface glycoconjugates from host target cells) by means of a unique trans-sialidase (TS) enzyme. TS-induced changes in the glycoprotein sialylation profile of both parasite and host cells are crucial for the establishment of a persistent T. cruzi infection and for the development of Chagas disease-associated pathogenesis. In this chapter, we describe a novel metabolic labeling method developed in our labs that enables straightforward identification and molecular characterization of SA acceptors of the TS-catalyzed reaction.