Co-transplantation of Xenogeneic Bone Marrow-derived Mesenchymal Stem Cells Alleviates Rejection of Pancreatic Islets in Non-obese Diabetic Mice.

Bone marrow-mesenchymal stem cells (BM-MSCs) have generated a great perspective in the field of regenerative medicine, and also in the treatment of inflammatory and autoimmune diseases in the past decade due to their immunomodulatory and anti-inflammatory properties. Here, we investigated the effect of xenogeneic BM-MSCs and pancreatic islets co-transplantation obtained from Wistar rats in preventing rejection or inducing tolerance to islet transplantation in non-obese diabetic mice. Non-obese diabetic mice were treated with co-transplantation of pancreatic islets and BM-MSCs (islet + MSCs group) or pancreatic islets only (islet group). Compared to the islet group, islet + MSCs had a lower expression of inflammatory markers, such as, tumor necrosis factor- α (13.40 ± 0.57 vs. 9.90 ± 0.12, P = .01), monocyte chemoattractant protein 1 (51.30 ± 6.80 vs. 9.00 ± 1.80, P = .01), and interleukin 1ß (IL-1ß) (16.2 ± 1.65 vs. 6.80 ± 1.00, P = .04). Comparing the expression of immune tolerance markers, it is noted that animals receiving the co-transplantation showed a significantly higher expression than the islet group of IL-4 (25.60 ± 1.96 vs. 2.80 ± 0.20, P = .004), IL-10 (188.40 ± 4.60 vs. 4.55 ± 0.12, P = .0001), and forkhead box P3 (34.20 ± 1.3 vs. 1.30 ± 0.2, P = .004), respectively. These results suggest an immunomodulatory action of BM-MSC in islet xenotransplantation showing that these stem cells have the potential to mitigate the early losses of grafts, due to the regulation of the inflammatory process of transplantation.

Dinâmica da população de coliformes durante a compostagem automatizada de dejetos líquidos de suínos / Population dynamics during composting of fecal automated pig slurry

Microrganismos presentes em dejetos de suínos podem contaminar o meio ambiente. Embora a compostagem seja preconizada como um método eficiente para reduzir este potencial poluidor dos dejetos, existem poucas informações de pesquisa sobre tal processo. O presente trabalho teve o objetivo de avaliar a eficiência da compostagem automatizada dos dejetos líquidos de suínos (DLS) na redução da população de coliformes, usados como indicadores de poluição fecal. Os DLS foram adicionados periodicamente, durante 106 dias, em substrato constituído pela mistura, em partes iguais, de maravalha e serragem. Foram efetuadas 14 adições de DLS, e em cada adição as leiras de compostagem eram revolvidas por meio de uma máquina especialmente desenvolvida para este fim. Foram avaliados dois tratamentos com três repetições, sendo um com e outro sem adição de ácido fosfórico aos dejetos, até pH 6,0. A adição de ácido visou reduzir as perdas de N por volatilização de amônia (NH3) durante a compostagem. A avaliação da população de coliformes foi feita pela técnica do número mais provável (NMP), com uso do caldo Fluorocult, incubado a 37ºC por 24h e posterior leitura em luz ultravioleta. A população de coliformes fecais não foi afetada pela adição de ácido fosfórico. O processo de compostagem automatizada foi eficiente na redução de coliformes fecais, cuja população original passou de 4,2x1010 para 1,2 x 105 ao final da compostagem (156 dias) sem adição de ácido e de 3,8x1010 para 2,3x104 na compostagem com adição de ácido. Essa remoção de coliformes fecais, promovida pela compostagem automatizada dos dejetos líquidos de suínos, corresponde a 99,99 por cento...

Microorganisms present in pig manure can contaminate the environment. Although composting is recommended as an efficient method to reduce the pollution potential of waste, there is little research information on this process. This study aimed to evaluate the efficiency of automated composting of pig slurry (PS) in reducing the population of coliforms, used as fecal pollution indicators. The PS was added periodically during 106 days in substrate, with a mixture, in equal parts, of wood shavings and sawdust. There were 14 additions of PS and at each addition the compost windrows were revolved through a machine especially developed for this purpose. Two treatments with three replications were evaluated, one with and one without the addition of phosphoric acid to the slurry up to pH 6.0. The acid addition aimed to reduce N losses through the volatilization of ammonia (NH3) during composting. Coliforms were evaluated by the technique of most probable number (MPN) using the Fluorocult broth, incubated at 37 ° C for 24 h and subsequent reading in ultra violet light. The population of fecal coliforms was not affected by the addition of phosphoric acid. The automated composting process was effective in reducing faecal coliforms, whose original population decreased from 4.2 x 1010 to 1.2 x 105 at the end of composting (156 days) without addition of acid and from 3.8 x1010 to 2,3 x104 in compost with added acid. This removal of faecal coliforms, promoted by automated composting of pig slurry, corresponds to 99.99 percent...

Expression of pancreatic endocrine markers by mesenchymal stem cells from human adipose tissue.

Mesenchymal stem cells (MSCs) from human adipose tissue have a great potential for use in cell therapy due to their ease of isolation, expansion, and differentiation, besides the relative acceptance from the ethical point of view. Our intention was to isolate and promote in vitro expansion and differentiation of MSCs from human adipose tissue into cells with a pancreatic endocrine phenotype. Human adipose tissue obtained from patients undergoing abdominal dermolipectomy was digested with type I collagenase. MSCs isolated by plastic adherence and characterized by cytochemistry and FACS were expanded in vitro. MSC differentiation into an endocrine phenotype was induced over 2 to 4 months with high glucose (25 mmol/L) media containing nicotinamide, exendin-4, and 2-mercaptoethanol. Insulin and glucagon expressions were analyzed by immunofluorescence. Cells isolated from human adipose tissue and expanded in vitro expressed MSC markers as confirmed by FACS and cytochemistry. Insulin but not glucagon production by differentiated cells was demonstrated by immunofluorescence. MSCs isolated from human adipose tissue were induced to differentiate in vitro into an endocrine phenotype that expressed insulin.

Expression of pancreatic endocrine markers by mesenchymal stem cells from human umbilical cord vein.

BACKGROUND: Mesenchymal stem cells (MSCs) from human umbilical cord vein have great potential for use in cell therapy because of their ease of isolation, expansion, and differentiation, in addition to their relative acceptance from the ethical point of view. Obtaining the umbilical cord at birth does not present any risk to either mother or child. OBJECTIVE: To isolate and promote in vitro expansion and differentiation of MSCs from human umbilical cord vein into cells with a pancreatic endocrine phenotype. METHODS: Mesenchymal stem cells obtained from human umbilical cord vein via collagenase digestion were characterized at cytochemistry and fluorescent-activated cell sorting, and expanded in vitro. Differentiation of MSCs into an endocrine phenotype was induced using high-glucose (23 mmol/L) medium containing nicotinamide, exendin-4, and 2-mercaptoethanol. Expression of insulin, somatostatin, glucagon, and pancreatic and duodenal homeobox 1 was analyzed using immunofluorescence. RESULTS: Cells isolated from the umbilical cord vein were MSCs as confirmed at cytochemistry and fluorescent-activated cell sorting. Expression of somatostatin, glucagon, and pancreatic and duodenal homeobox 1 by differentiated cells was demonstrated using immunofluorescence. Insulin was not expressed. CONCLUSIONS: The MSC differentiation protocol used in the present study induced expression of some endocrine markers. Insulin was not produced by these cells, probably because of incomplete induction of differentiation.

Expression of pancreatic endocrine markers by prolactin-treated rat bone marrow mesenchymal stem cells.

BACKGROUND: Mesenchymal stem cells (MSCs) are an attractive source for generation of cells with beta-cell properties. Previous studies have demonstrated the ability of prolactin to induce an increase in beta-cell mass and maturation, which suggests beneficial effects of its use in MSC differentiation protocols. OBJECTIVE: To evaluate the expression of endocrine differentiation markers in rat MSCs treated in vitro with prolactin. METHODS: Mesenchymal stem cells from bone marrow of Wistar rats were isolated, expanded, and characterized. Differentiation of MSCs was induced in medium containing 23 mmol/L of glucose, and nicotinamide, 2-mercaptoethanol, and exendin-4, in the presence or absence of 500 ng/mL of rat recombinant prolactin. Expression of endocrine markers and prolactin receptor genes was evaluated using real-time polymerase chain reaction, and compared between culture stages and presence vs absence of prolactin in the culture medium. Expression of insulin, somatostatin, glucagon, and pancreatic and duodenal homeobox 1 was also evaluated at immunofluorescence microscopy. RESULTS: Isolated cells were mostly MSCs, as confirmed at fluorescent-activated cell sorting and cytochemistry. Pax6, Ngn-3, Isl1, NeuroD1, Nkx2.2, and Nkx6.1 exhibited varied expression during culture stages. The long form of the prolactin receptor messenger RNA was induced in prolactin-treated cultures (P < .05). The somatostatin gene was induced in early stages of differentiation (P < .05), and its expression was induced by prolactin, as confirmed using immunofluorescence. CONCLUSION: Culture of rat bone marrow MSCs in differentiation medium induces expression of pancreatic endocrine-specific genes, and somatostatin and prolactin receptor expression was also induced by prolactin.

Increased plasma and endothelial cell expression of chemokines and adhesion molecules in chronic kidney disease.

Chemokines and adhesion molecules are involved in early events of atherogenesis. In the present study, we investigated the effects of the uremic milieu on the expression of monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8), soluble vascular adhesion molecule-1 (sVCAM-1) and soluble intercellular adhesion molecule-1 (sICAM-1) and their relationship to cardiovascular status. Plasma samples were obtained from patients in different stages of chronic kidney disease (CKD). Cardiovascular status was evaluated by intima-media thickness and endothelial dysfunction by flow mediation dilatation and proteinuria. In vitro studies were performed using human umbilical endothelial cells exposed to uremic plasma or plasma from healthy subjects. MCP-1, IL-8, sVCAM-1 and sICAM-1 levels in plasma and in supernatant were analyzed by enzyme-linked immunosorbent assay. The population consisted of 73 (mean age 57 years; 48% males) CKD patients with glomerular filtration rate (GFR) of 37 +/- 2 ml/min. MCP-1 and sVCAM-1 plasma levels were negatively correlated with GFR (rho = -0.40, p < 0.0005 and rho = -0.42, p < 0.0005, respectively). Fibrinogen was positively correlated with MCP-1, sICAM-1 and sVCAM-1 (rho = 0.33, p < 0.005, rho = 0.32, p < 0.05 and rho = 0.25, p < 0.05, respectively) and ultra-high-sensitivity C-reactive protein was positively correlated with sICAM-1 (rho = 0.25, p < 0.0005). Plasma IL-8 had a significant positive correlation with proteinuria (rho = 0.31, p < 0.01). There was a time- and CKD-stage-dependent MCP-1, IL-8 and sVCAM-1 endothelial expression (p < 0.05). In summary, plasma levels of markers of endothelial cell activation (MCP-1 and sVCAM-1) are increased in more advanced CKD. Exposure of endothelial cells to uremic plasma results in a time- and CKD-stage-dependent increased expression of MCP-1, IL-8 and sVCAM-1, suggesting a link between vascular activation, systemic inflammation and uremic toxicity. Future studies are necessary to investigate whether these biomarkers add predictive value in comparison to the previously described ones. Also, endothelial response to uremic toxicity should be viewed as a potential target for intervention in order to reduce morbidity and mortality in CKD-related cardiovascular disease.

Hematoma subcapsular hepático por fasciolasis / Hepatic subcapsular hematoma caused by fascioliasis

Hepatic fasciolasis is a worldwide spread zoonoses mainly affecting cattle-raising countries. It is caused by the trematode Fasciola hepßtica and it is characterized by abdominal pain, fever, nausea and vomitus, weight loss, diahrrea, paleness, general malaise, and hypereosinophilia. Immunological diagnosis as well as stool eggs count may be performed. Hepatic subcapsular and intraparenchymatous hematoma is an infrequent complication of human fascioliasis. Nevertheless, for establishing a proper diagnosis and treatment, any suspicion of its presence must be carefully discarded through, clinical epidemiology, laboratory and imaging exams and procedures. The aim of this study is to expand knowledge on this unfrequently dealt pathology in medical literature by presenting four case reports related to patients undergoing a two-year treatment. All of them had been referred from Departamento de Cajamarca, Peru.

La fasciolasis hepática es una zoonosis mundialmente difundida, sobre todo en los países productores de ganado; causada por la fasciola hepática. Se manifiesta por dolor abdominal, fiebre, nauseas y vómitos, baja de peso, diarrea, palidez, malestar general e hipereosinofilia. El diagnóstico es inmunológico y también puede hacerse por recuento de huevos en heces. El hematoma subcapsular e intraparenquimatoso hepático es una complicación rara de la fasciolasis humana pero se debe tener un alto índice de sospecha uniendo epidemiología, clínica, laboratorio e imaginologia para un adecuado diagnóstico y tratamiento. El propósito de reportar estos casos es dar a conocer una patología poco frecuente en la literatura, con una casuística de cuatro pacientes tratados en el lapso de dos años, todos referidos del Departamento de Cajamarca.

Microencapsulation and tissue engineering as an alternative treatment of diabetes

In the 70's, pancreatic islet transplantation arose as an attractive alternative to restore normoglycemia; however, the scarcity of donors and difficulties with allotransplants, even under immunosuppressive treatment, greatly hampered the use of this alternative. Several materials and devices have been developed to circumvent the problem of islet rejection by the recipient, but, so far, none has proved to be totally effective. A major barrier to transpose is the highly organized islet architecture and its physical and chemical setting in the pancreatic parenchyma. In order to tackle this problem, we assembled a multidisciplinary team that has been working towards setting up the Human Pancreatic Islets Unit at the Chemistry Institute of the University of São Paulo, to collect and process pancreas from human donors, upon consent, in order to produce purified, viable and functional islets to be used in transplants. Collaboration with the private enterprise has allowed access to the latest developed biomaterials for islet encapsulation and immunoisolation. Reasoning that the natural islet microenvironment should be mimicked for optimum viability and function, we set out to isolate extracellular matrix components from human pancreas, not only for analytical purposes, but also to be used as supplementary components of encapsulating materials. A protocol was designed to routinely culture different pancreatic tissues (islets, parenchyma and ducts) in the presence of several pancreatic extracellular matrix components and peptide growth factors to enrich the beta cell population in vitro before transplantation into patients. In addition to representing a therapeutic promise, this initiative is an example of productive partnership between the medical and scientific sectors of the university and private enterprises.

Microencapsulation and tissue engineering as an alternative treatment of diabetes.

In the 70's, pancreatic islet transplantation arose as an attractive alternative to restore normoglycemia; however, the scarcity of donors and difficulties with allotransplants, even under immunosuppressive treatment, greatly hampered the use of this alternative. Several materials and devices have been developed to circumvent the problem of islet rejection by the recipient, but, so far, none has proved to be totally effective. A major barrier to transpose is the highly organized islet architecture and its physical and chemical setting in the pancreatic parenchyma. In order to tackle this problem, we assembled a multidisciplinary team that has been working towards setting up the Human Pancreatic Islets Unit at the Chemistry Institute of the University of São Paulo, to collect and process pancreas from human donors, upon consent, in order to produce purified, viable and functional islets to be used in transplants. Collaboration with the private enterprise has allowed access to the latest developed biomaterials for islet encapsulation and immunoisolation. Reasoning that the natural islet microenvironment should be mimicked for optimum viability and function, we set out to isolate extracellular matrix components from human pancreas, not only for analytical purposes, but also to be used as supplementary components of encapsulating materials. A protocol was designed to routinely culture different pancreatic tissues (islets, parenchyma and ducts) in the presence of several pancreatic extracellular matrix components and peptide growth factors to enrich the beta cell population in vitro before transplantation into patients. In addition to representing a therapeutic promise, this initiative is an example of productive partnership between the medical and scientific sectors of the university and private enterprises.