Endo Belly: What Is It and Why Does It Happen?—A Narrative Review

Endometriosis is a chronic inflammatory disease where endometrial-like lesions settle outside the uterus, resulting in extensive inflammatory reactions. It is a complex disease that presents with a range of symptoms, with pain and infertility being the most common. Along with severe dysmenorrhea, cyclic and acyclic lower abdominal pain, cyclic dysuria and dyschezia, dyspareunia, and infertility, there are also nonspecific complaints that can cause confusion and make endometriosis the chameleon among gynecological diseases. These symptoms include unspecific intestinal complaints, cyclic diarrhea, but also constipation, nausea, vomiting, and stomach complaints. It appears that in addition to general bowel symptoms, there are also specific symptoms related to endometriosis such as cyclic bloating of the abdomen, known as endo belly. During the second half of the menstrual cycle leading up to menstruation, the abdomen becomes increasingly bloated causing discomfort and pain due to elevated sensitivity of the intestinal wall. Patients with endometriosis exhibit a reduced stretch pain threshold of the intestinal wall. Here, we review the endo belly, for the first time, pathophysiology and the influence of other diseases (such as irritable bowel syndrome-IBS), microbiome, hormonal levels, inflammation, and diet on the presentation of this condition

Neurogenic Inflammation in the Context of Endometriosis—What Do We Know?

Endometriosis (EM) is an estrogen-dependent disease characterized by the presence of epithelial, stromal, and smooth muscle cells outside the uterine cavity. It is a chronic and debilitating condition affecting ~10% of women. EM is characterized by infertility and pain, such as dysmenorrhea, chronic pelvic pain, dyspareunia, dysuria, and dyschezia. Although EM was first described in 1860, its aetiology and pathogenesis remain uncertain. Recent evidence demonstrates that the peripheral nervous system plays an important role in the pathophysiology of this disease. Sensory nerves, which surround and innervate endometriotic lesions, not only drive the chronic and debilitating pain associated with EM but also contribute to a growth phenotype by secreting neurotrophic factors and interacting with surrounding immune cells. Here we review the role that peripheral nerves play in driving and maintaining endometriotic lesions. A better understanding of the role of this system, as well as its interactions with immune cells, will unearth novel disease-relevant pathways and targets, providing new therapeutics and better-tailored treatment options

In vitro substrate reduction, chaperone and immunomodulation treatments reduce heparan sulfate in mucolipidosis III human fibroblasts

Abstract Mucolipidosis II and III (MLII and MLIII) are autosomal recessive diseases caused by pathogenic variants in GNPTAB and GNPTG genes that lead to defects in GlcNAc-1-phosphotransferase. This enzyme adds mannose 6-phosphate residues to lysosomal hydrolases, which allows enzymes to enter lysosomes. Defective GlcNAc-1-phosphotransferase causes substrate accumulation and inflammation. These diseases have no treatment, and we hypothesized that the use of substrate reduction therapy and immunomodulation may be beneficial at the cell level and as a future therapeutic approach. Fibroblasts from two patients with MLIII alpha/beta and 2 patients with MLIII gamma as well as from one healthy control were treated with 10 µM miglustat, 20 µM genistein, and 20 µM thalidomide independently. ELISA assay and confocal immunofluorescence microscopy were used to evaluate the presence of heparan sulfate (HS) and the impact on substrate accumulation. ELISA assay showed HS reduction in all patients with the different treatments used (p=0.05). HS reduction was also observed by immunofluorescence microscopy. Our study produced encouraging results, since the reduction in substrate accumulation, even partial, may offer benefits to the phenotype of patients with inborn errors of metabolism

Production of Proteolytic Enzymes by a Keratin-Degrading Aspergillus niger

A fungal isolate with capability to grow in keratinous substrate as only source of carbon and nitrogen was identified as Aspergillus niger using the sequencing of the ITS region of the rDNA. This strain produced a slightly acid keratinase and an acid protease during cultivation in feather meal. The peak of keratinolytic activity occurred in 48 h and the maximum proteolytic activity in 96 h. These enzymes were partly characterized as serine protease and aspartic protease, respectively. The effects of feather meal concentration and initial pH on enzyme production were evaluated using a central composite design combined with response surface methodology. The optimal conditions were determined as pH 5.0 for protease and 7.8 for keratinase and 20 g/L of feather meal, showing that both models were predictive. Production of keratinases by A. niger is a less-exploited field that might represent a novel and promising biotechnological application for this microorganism

Humoral immune response in adult Brazilian patients with mucolipidosis III gamma

Mucolipidosis II and III (ML II and III) alpha/beta and ML III gamma are lysosomal diseases caused by GlcNAc-1-phosphotransferase deficiency. Previous data indicate that MLII patients have functionally impaired immune system that contributes to predisposition to infections.We evaluated the immunological phenotype of three Brazilian patients with ML III gamma. Our data suggest that the residual activity of GlcNAc-1-phosphotransferase in patients with ML III gamma is enough to allow the targeting of the lysosomal enzymes required for B-cell functions maintenance

AVALIAÇÃO DA ATIVIDADE ANTIMICROBIANA DO ÓLEO VOLÁTIL E EXTRATOS ETANÓLICOS DE FOLHAS E RAMOS DE ILEX PARAGUARIENSIS A. ST.-HIL. (ERVA-MATE)

Among the many species of Brazilian native flora is Ilex paraguariensis, popularly known as erva-mate. Studies attribute several biological activities to this species. The objective of this work was to evaluate the antifungal and antibacterial activity of the volatile oil, extract of the leaves and the branches of the I. paraguariensis. The plant material was harvested at URI (28 ° 16’41.4 “S 54 ° 16’13.0” W) in Santo Ângelo - RS / Brazil, being ground and filtered and the volatile oil obtained by hydrodistillation. The antimicrobial activity was evaluated using the Brain Heart infusion (BHI) solid medium. The bacteria used were Corynebacterium fimi, Listeria monocytogenes, Bacillus cereus, Staphylococcus aureus, Salmonella enteritidis and Escherichia coli. For antifungal activity the method of diffusion in agar medium Patato dextrose agar (PDA) was used. The fungi used were Aspergillus phoenicis isolated, Aspergillus niger, Fusarium sp. and Fusarium sp. isolated clinic. The aliquots employed were the same for both bacteria and fungi of 25 and 50 μL at the concentrations of 50 and 100 mg / mL of oil and extracts. The results obtained, when compared to the results of inhibition of Fox® commercial antifungal, used as a comparative standard, proved the marked antimicrobial activity of I. paraguariensis oil. Among the different extracts tested, the leaves showed better results for antimicrobial activity with the concentration of 100mg /mL of the oil. The volatile oil of I. paraguariensis, due to the presence of inhibition halos greater than 10 mm indicates a good antimicrobial activity.Dentre as inúmeras espécies da flora nativa brasileira, encontra-se a Ilex paraguariensis, conhecida popularmente como erva-mate. Estudos atribuem diversas atividades biológicas para essa espécie. O trabalho teve por objetivo avaliar a atividade antifúngica e antibacteriana do óleo volátil, extrato das folhas e dos ramos da I. paraguariensis. O material vegetal foi coletado na URI (28°16’41.4”S 54°16’13.0”W) em Santo Ângelo – RS/Brasil, sendo moído e filtrado e o óleo volátil obtido por hidrodestilação. A atividade antimicrobiana foi avaliada empregando a técnica de difusão em meio sólido Brain heart infusion (BHI). As bactérias utilizadas foram Corynebacterium fimi, Listeria monocytogenes, Bacillus cereus, Staphylococcus aureus, Salmonella enteritidis e Escherichia coli. Para a atividade antifúngica, foi empregado o método de difusão em meio sólido ágar Patato dextrose (PDA), e os fungos utilizados foram Aspergillus phoenicis isolado, Aspergillus niger, Fusarium sp. e Fusarium sp. isolado clínico. As alíquotas empregadas foram as mesmas, tanto para as bactérias quanto para os fungos, de 25 e 50 µL nas concentrações de 50 e 100 mg/mL do óleo e extratos. Os resultados obtidos, quando comparados aos resultados de inibição do antifúngico comercial Fox®, usados como padrão comparativo, comprovaram a acentuada atividade antimicrobiana apresentada pelo óleo de I. paraguariensis. Entre os diferentes extratos testados, o de folhas apresentou melhor resultado para atividade antimicrobiana, com a concentração de 100 mg/mL do óleo. O óleo volátil de I. paraguariensis, por apresentar halos de inibição superiores a 10 mm, indica uma boa atividade antimicrobiana

ARSB IS REQUIRED FOR BONE REMODELING

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Estudo abrangente sobre a deficiência da GlcNac-1-fosfotransferase (Mucolipidoses II e III) : do diagnóstico molecular a propostas de tratamento

Introdução: Os lisossomos são organelas ácidas em que muitos tipos de macromoléculas, incluindo proteínas, carboidratos, ácidos nucléicos e lipídios são entregues para a degradação. A biogênese dos lisossomos requer a contínua substituição das enzimas lisossomais solúveis e das proteínas de membrana lisossomal. O direcionamento das hidrolases ácidas depende dos resíduos de manose-6-fosfato (M6P) que são reconhecidos por receptores específicos que medeiam o seu transporte para os lisossomos. O papel chave na formação destes resíduos é desempenhado no Complexo de Golgi pela enzima GlcNAc-1-fosfotransferase que contêm seis subunidades, α2β2γ2. Este complexo enzimático é codificado por dois genes, GNPTAB e GNPTG. Mutações nestes genes resultam em duas doenças lisossômicas, as mucolipidoses (ML) tipo II e III, caracterizadas bioquimicamente pela perda de múltiplas enzimas lisossomais, devido à formação deficiente dos resíduos de M6P. Objetivos: 1) Verificar a patogenicidade, por meio de ensaios in vitro, das seguintes mutações em GNPTAB previamente identificadas em pacientes brasileiros com ML II e III alfa/beta: c.1208T>C, c.1514G>A, c.1723G>A, c.1759C>T, c.1931_1932CA>TG, c.2269_2273delGAAAC, c.2808A>G e c.3668_3670delCTA; 2) Caracterizar o perfil de mutações de GNPTG presente em uma amostra de pacientes com ML III gama; 3) Analisar o efeito de gentamicina e cloranfenicol sobre a atividade das enzimas α-manosidase, β-glicuronidase e β-galactosidase em fibroblastos de pacientes com ML III gama heterozigotos ou homozigotos para mutações sem sentido em GNPTG. Metodologia: Estudo transversal, amostragem por conveniência, incluindo pacientes com diagnóstico clínico e bioquímico de ML II/III. Mutações em GNPTAB previamente identificadas em pacientes brasileiros com ML II/III alfa/beta foram estudas em relação a expressão de mRNA, proteína, atividade enzimática e localização intracelular da GlcNAc-1-fosfotransferase. Pacientes também foram avaliados quanto a variações no gene GNPTG e seus possíveis impactos a nível de mRNA. O grupo de pacientes com ML III gama foram avaliados para medir sua independência funcional “Functional Independence Measure” (FIM). Estudos de expressão de GNPTAB e GNPTG também foram desenvolvidos para maior compreensão da relação destes genes. O desenvolvimento de um possível tratamento realizado nos fibroblastos de pacientes com ML III gama também foram objeto de estudo. Resultados: Mutações sem sentido e que alteram a fase de leitura da enzima GlcNAc-1-fosfotransferase não são corretamente transportados do retículo endoplasmático (RE) ao aparelho de Golgi, devido a interrupção, a falta dos sinais de exportação localizados nas porções N- e C- terminais da proteína. O não transporte ao Golgi e consequentemente, a ausência da clivagem proteolítica da proteína precursora das subunidades α/β, são responsáveis pela falta de atividade enzimática. Além dessas, mutações com sentido trocado na porção luminal da enzima também podem ter o transporte ao Golgi prejudicado, o que sugere a presença de um local de contato para uma proteína necessária para a exportação eficiente do RE. O estabelecimento de um ensaio radioativo para medir a atividade enzimática da GlcNAc-1-fosfotransferase confirmou que o transporte para o Golgi e a clivagem proteolítica nas subunidades α- e β- maduras é um pré-requisito para a atividade enzimática. As mutações em GNPTAB tiveram sua patogenicidade confirmada. Em relação a análise de GNPTG, as técnicas moleculares empregadas permitiram a identificação de três novas mutações patogênicas (c.244_247dupGAGT, c.328G>T e c.233+5G>C) e de uma outra variação (c.-112C>G). Após incessante investigação do trio cuja mutação causal c.244_247dupGAGT foi encontrada, a mesma foi atribuída como um evento de novo que ocorreu em um único óvulo ou de um mosaicismo germinativo no óvulo materno. Ao investigar o potencial efeito das mutações sem sentido através de mRNA, não foi possível identificar a mutação c.328G>T (p.E110X), no lugar desta, todos os pacientes apresentaram a sequência selvagem de GNPTG, evento denominado edição de RNA (c.328G@T). Porém, níveis de mRNA muito próximos a zero foram obtidos o que confirma a patogenicidade das mutações em GNPTG bem como o diagnóstico dos pacientes. Em pacientes ML II e III alfa/beta e em ML III gama, a redução dos níveis de mRNA de GNPTAB e GNPTG, respectivamente, pode ser facilmente explicada pela natureza das mutações identificadas nos pacientes. O que intriga, porém, é a expressão do gene não mutado. A divergência entre os resultados encontrados pode estar relacionada ao tipo de amostra, sabendo-se que as ML II e ML III são tecido-específicas. Fibroblastos de três pacientes com ML III gama portadores de mutações sem sentido foram tratados com geneticina e cloranfenicol. Este estudo de conceitos não demonstrou a possibilidade ou eficácia de um tratamento ser desenvolvido baseado na utilização de compostos não antibióticos atuarem sobre a tradução alternativa ou read through em pacientes com ML II e III. Conclusão: Estudos de caracterização genética, clínica e populacional sempre contribuirão para a elucidação dos mecanismos da doença e concomitantemente, melhor atendimento ao paciente. Com isto, é de grande importância que esforços e recursos sejam destinados a pesquisas nesta área para o completo entendimento das ML II e III e dos processos biológicos envolvidos. Este é o primeiro estudo brasileira a realizar o diagnóstico molecular de ML III gama; o primeiro a relatar uma mutação de novo e também, a ocorrência de edição de mRNA em pacientes com ML III gama.Introduction: Lysosomes are acidic organelles into which many types of macromolecules including proteins, carbohydrates, nucleic acids and lipids are delivered for degradation. The biogenesis of lysosomes requires a continuous substitution of soluble lysosomal enzymes and lysosomal membrane proteins. The targeting of lysosomal enzymes depends on mannose 6-phosphate residues (M6P) that are recognized by M6P-specific receptors mediating their transport to lysosomes. The key role in the formation of M6P residues plays the Golgi-resident GlcNAc-1-phosphotransferase complex consisting of six subunits, α2β2γ2. This enzyme complex is encoded by two genes, GNPTAB and GNPTG. Mutations in these genes result in two lysosomal storage diseases, mucolipidosis (ML) type II and III, biochemically characterized by the missorting of multiple lysosomal enzymes due to impaired formation of M6P residues, and general lysosomal dysfunction. Objectives: 1)To analyze the pathogenicity, through in vitro tests, of GNPTAB mutations previously identified in Brazilian patients with ML II and III alpha/beta: c.1208T>C, c.1514G>A, c.1723G>A, c.1759C>T, c.1931_1932CA>TG, c.2269_2273delGAAAC, c.2808A>G and c.3668_3670delCTA; 2) Characterize GNPTG mutational profile in a ML III gamma patients group; 3) Analyze the effect of gentamicin and chloramphenicol on the activity of α-mannosidase, β-glucuronidase and β-galactosidasein fibroblasts of ML III gamma patients with GNPTG nonsense mutations. Methods: Cross-sectional study, with convenience sample, including patients with a clinical and biochemical diagnosis of ML II/III. GNPTAB mutations previously identified in ML II/III alpha/beta Brazilian patients were studied in relation to mRNA and protein expression, enzyme activity and intracellular localization of GlcNAc-1-phosphotransferase. Patients were also evaluated to GNPTG variations and possible mRNA impacts. The ML III gamma patients group was also evaluated with the Functional Independence Measure (FIM). GNPTAB and GNPTG expression studies were developed to better understand the relationship between them. Geneticin and chloramphenicol were used to treat ML III gamma fibroblasts. Results: Nonsense and frameshift mutations of GlcNAc-1-phosphotransferase failed to reach the Golgi apparatus due to the interrupted cooperative ER export signals localized both in the N- and C-terminal cytosolic tails and lacked proteolytic activation of the α/β-subunit precursor. In addition, luminal missense mutations of the GlcNAc-1-phosphotransferase can also impair the transport to the Golgi apparatus, suggesting the presence of a protein contact site required for efficient ER export. The establishment of a radioactive assay to measure the activity of the GlcNAc-1-phosphotransferase confirmed that the transport to the Golgi and proteolytic cleavage into mature α- and β-subunits is prerequisite for enzymatic activity. Regarding GNPTG analysis, molecular techniques employed allowed the identification of three new pathogenic mutations (p.F83X, p.E110X, c.233+5G>C) and another variation (c.-112C>G). After research, p.F83X was attributable to a de novo event which occurred in only one ovum, or to germline mosaicism in the mothers’ ova. The potential effect of p.E110X mutation in mRNA was investigated. However it was not possible to identify transcripts carrying this mutation since all patients appear to present only the wild type sequence, an event called mRNA editing (c.328G@T). Nevertheless, low mRNA levels confirm the GNPTG mutations pathogenicity and the patients’ diagnosis. In ML II/III alpha/beta and ML III gamma patients, low GNPTAB and GNPTG mRNA expression levels, respectively, can easily be explained by the nature of the mutations. Interestingly, it is the non-mutated gene. Divergence between results may be related to the type of sample, given that ML II and III are tissue-specific diseases. Fibroblasts from three ML III gamma patients were treated with geneticin and chloramphenicol with no effect being observed. This pilot study does not support the feasibility or effectiveness for the development of a treatment based on the use of non-antibiotic compounds acting on the read through of either ML II or III patients. Conclusions: Genetic, clinical and population characterization studies always contribute to the elucidation of disease mechanisms and concomitantly, lead to better patient care. It is very important that efforts and resources to be focused in this area for the complete understanding of ML II and III as well as the biological processes involved. This is the first study to perform molecular diagnosis in ML III gamma Brazilian patients. Also it is the first to report a de novo mutation and the occurrence of mRNA editing in ML III gamma patients