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Aim : In this study, we evaluated the impact of an integrated oral care protocol for pediatric patients undergoing antineoplastic treatment from the perspective of the multidisciplinary team, oral healthcare team, and caregivers. Subject and Methods: This was a qualitative study carried out in the pediatric sector of a reference cancer hospital in Brazil. Focus group and individual semi-structured interview techniques were used, and the data were analyzed by the Discourse of the Collective Subject (DCS) method. Results: A total of 44 professionals involved in providing care to children and adolescents with cancer and 38 caregivers were interviewed. All interviewees perceived an improvement in the patients' oral health condition. Health professionals reported a reduction in the occurrence and severity of oral mucositis (OM). Communicating with the medical team and understanding the importance of oral care to the patient's systemic condition were the greatest difficulties reported by the oral healthcare team. Conclusion: This implementation project contributed to establishing a complete multidisciplinary team to assist pediatric patients in all their needs during antineoplastic treatment. The integrated oral care protocol further contributed to reducing the occurrence and severity of OM by increasing its surveillance and diagnostic efficiency, which, altogether, improved the patients' quality of life. [ABSTRACT FROM AUTHOR]

Bone marrow cells (BMCs) from obese Swiss mice fed with Western diet show mitochondrial dysfunction. Obesity interferes with BMCs disrupting energetic metabolism, stimulating apoptosis, and reducing cell proliferation since adipose tissue releases inflammatory adipokines into the medullar microenvironment. These changes lead to reduction of BMC differentiation capacity and hematopoiesis impairment, a process responsible for blood cell continuous production through hematopoietic stem cells (HSCs). This work aimed to analyze the effects of IGF-1 therapy on BMC viability in Western diet-induced obesity, in vivo. We observed that after only 1 week of treatment, obese Swiss mice presented reduced body weight and visceral fat and increased mitochondrial oxidative capacity and coupling, indicating mitochondrial function improvement. In addition, IGF-1 was able to reduce apoptosis of total BMCs, stem cell subpopulations (hematopoietic and mesenchymal), and leukocytes, restoring all progenitor hematopoietic lineages. The treatment also contributed to increase proliferative capacity of hematopoietic stem cells and leukocytes, keeping the hematopoietic and immune systems balanced. Therefore, we conclude that IGF-1 short period therapy improved BMC survival, proliferation, and differentiation capacity in obese Swiss mice. [ABSTRACT FROM AUTHOR]

For medical use of proteins and peptide-based drugs, it is desirable to have small biologically active sequences because they improve stability, reduce side effects, and production costs. Several plant defensins have their biological activities imparted by a sequence named γ-core. Vu-Def, a Vigna unguiculata defensin, has activity against Leishmania amazonensis, which is one etiological agent of leishmaniasis and for which new drugs are needed. Our intention was to understand if the region comprising the Vu-Def γ-core is responsible for the biological activity against L. amazonensis and to unveil its mechanism of action. Different microbiological assays with L. amazonensis in the presence of the synthetic peptide A36,42,44γ32-46Vu-Def were done, as well as ultrastructural and fluorescent analyses. A36,42,44γ32-46Vu-Def showed biological activity similar to Vu-Def. A36,42,44γ32-46Vu-Def (74 µM) caused 97% inhibition of L. amazonensis culture and parasites were unable to regrow in fresh medium. The cells of the treated parasites showed morphological alterations by ultrastructural analysis and fluorescent labelings that corroborate with the data of the organelles alterations. The general significance of our work is based on the description of a small synthetic peptide, A36,42,44γ32-46Vu-Def, which has activity on L. amazonensis and that the interaction between A36,42,44γ32-46Vu-Def-L. amazonensis results in parasite inhibition by the activation of an apoptotic-like cell death pathway. [ABSTRACT FROM AUTHOR]

Trypanosoma cruzi is an obligate intracellular parasite transmitted to vertebrate hosts by blood-sucking insects. Molecules present in parasites and mammalian cells allow the recognition and parasite internalization. Metallic ions play an essential role in the establishment and maintenance of host-parasite interaction. However, little is known about how parasites handle with essential and nonessential metal quotas. This study aimed to investigate the influence of metal ions on the biological processes of T. cruzi infected cells. Infected cells were incubated with ZnCl2, CdCl2, and HgCl2 for 12 h and labeled with different specific dyes to investigate the cellular events related to intracellular parasite death and elimination. Infected host cells and parasite's mitochondria underwent functional and structural disorders, in addition to parasite's DNA condensation and pH decrease on host cells, which led to parasite death. Further investigations suggested that lysosomes were involved in pH decrease and the double membrane of the endoplasmic reticulum formed vacuoles surrounding damaged parasites, which indicate the occurrence of autophagy for parasite elimination. In conclusion, low concentrations of nonessential and essential metals cause a series of damage to Trypanosoma cruzi organelles, leading to its loss of viability, death, and elimination, with no removal of the host cells. [ABSTRACT FROM AUTHOR]

Trypanosoma cruzi is transmitted to vertebrate hosts during the feeding of blood-sucking insects. After the invasion of host cells, the parasite resides within the parasitophorous vacuole until to escape to host cytoplasm and to proliferate, establishing an infection. Studies demonstrated that some intracellular parasites have to acquire all essential nutrients as well as transition metals from the host cell to be pathogenic, to maintain the homeostasis and to replicate. The present study investigated the progressive steps of the intracellular parasite development and establishment of infection in the presence of ZnCl2, CdCl2 and HgCl2. LLC-MK2 cells were infected with trypomastigotes during 6-84 h to investigate the steps of intracellular parasite development. After the host cells were infected during 12 h and treated with metals during 24 or 60 h or they were treated for 24 h and cultured for 72 h more to observe the reversibility. The results showed that the non-synchronous invasion of trypomastigotes resulted in an increasing number of intracellular parasites in intermediary forms (until 24 h post-infection), the appearance (from 36 h) and proliferation (84 h) of the amastigotes. The 24 h-treatments were not enough to impair parasite escape to the host cytoplasm and reproduction. However, 60 h of incubations led to a significant reduction in parasite numbers, as well as the reversibility assays. In conclusion, new insights about the intracellular T. cruzi development in the presence of metals were provided, and further studies should be performed to investigate the events involved in parasite death and elimination. [ABSTRACT FROM AUTHOR]

Nutritional changes in the development (intrauterine life and postnatal period) may trigger long-term pathophysiological complications such as obesity and cardiovascular disease. Metabolic programming leads to organs and tissues modifications, including adipose tissue, with increased lipogenesis, production of inflammatory cytokines, and decreased glucose uptake. However, stem cells participation in adipose tissue dysfunctions triggered by overfeeding during lactation has not been elucidated. Therefore, this study was the first to evaluate the effect of metabolic programming on adipose mesenchymal stem cells (ASC) from mice submitted to overfeeding during lactation, using the litter reduction model. Cells were evaluated for proliferation capacity, viability, immunophenotyping, and reactive oxygen species (ROS) production. The content of UCP-2 and PGC1-α was determined by Western Blot. ASC differentiation potential in adipogenic and osteogenic environments was also evaluated, as well the markers of adipogenic differentiation (PPAR-γ and FAB4) and osteogenic differentiation (osteocalcin) by RT-qPCR. Results indicated that neonatal overfeeding does not affect ASC proliferation, ROS production, and viability. However, differentiation potential and proteins related to metabolism were altered. ASC from overfed group presented increased adipogenic differentiation, decreased osteogenic differentiation, and also showed increased PGC1-α protein content and reduced UCP-2 expression. Thus, ASC may be involved with the increased adiposity observed in neonatal overfeeding, and its therapeutic potential may be affected. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii, the etiological agent of toxoplasmosis, infects nucleated cells and then resides and multiplies within a parasitophorous vacuole. For this purpose, the parasite secretes many virulence factors for the purpose of invading and subverting the host microbicidal defenses in order to facilitate its survival in the intracellular milieu. Essential metals are structural components of proteins and enzymes or cofactors of enzymatic reactions responsible for these parasitic survival mechanisms. However, an excess of non-essential or essential metals can lead to parasite death. Thus, infected host cells were incubated with 20 µM ZnCl2 in conjunction with 3 µMCdCl2 or HgCl2 for 12 h in order to investigate cellular events and organelle damage related to intracellular parasite death and elimination. In the presence of these metals, the tachyzoites undergo lipid uptake and transport impairment, functional and structural mitochondrial disorders, DNA condensation, and acidification of the parasitophorous vacuole, thus leading to parasite death. Additional research has suggested that lysosome-vacuole fusion was involved in parasite elimination since acid phosphatases were found inside the parasitophorous vacuole, and vacuoles containing parasites were also positive for autophagy. In conclusion, low concentrations of CdCl2, HgCl2, and ZnCl2 can cause damage to Toxoplasma gondii organelles, leading to loss of viability, organelle death, and elimination without causing toxic effects to host cells. [ABSTRACT FROM AUTHOR]

Trypanosoma cruzi has many molecules that need metallic elements to work, allowing cell invasion and the establishment of infection, causing Chagas disease. Nonetheless, knowledge regarding how the parasites address metals and maintain homeostasis is lacking. To study this relationship, zinc, cadmium and mercury were chosen. Epimastigote, trypomastigote and intracellular forms of T. cruzi were incubated with these metals for different times and at different concentrations. In general, epimastigotes were the most sensitive and trypomastigotes the most resistant to metals. ZnCl induced low toxic effects to all parasite forms. Although the parasites were very sensitive to the toxic effects of CdCl and HgCl, pretreatment with ZnCl decreased the death rate. The trypomastigotes pretreated with CdCl were unable to infect the host cells, and the treated intracellular forms were damaged after 2 h of incubation, when the toxic effects were poorly reverted. New insights on metal toxicity mechanisms are provided, helping to understand how metallic ions influence the parasite's biochemical and physiological processes. [ABSTRACT FROM AUTHOR]

Renovascular hypertension (RVH) is a progressive disease, leading to chronic kidney disease when untreated and no specific treatment is available. Therefore, development of new therapeutic modalities is imperative. RVH is triggered by renal artery stenosis and subsequent renin-angiotensin-aldosterone system activation; it can be experimentally induced by the 2 Kidneys-1 Clip (2K1C) model. This study investigates the therapeutic potential of renal subcapsular mesenchymal stem cell (MSC) infusion in 2K1C rats. Renal morphological and functional changes were analyzed, including Na+K-ATPase activity and expression, renin angiotensin-converting enzyme (ACE) and angiotensin-II type 1 (ATR) and type 2 (ATR) receptors expression. 2K1C rats developed hypertension accompanied by renin upregulation (clipped kidney) and renal Na+K-ATPase activity and expression reduction. MSC therapy decreased systolic blood pressure, renin, ACE, and ATR, upregulated ATR and podocin expression and restored renal Na+K-ATPase activity and expression. In addition, MSC improved renal morphology, reduced fibrosis and TGF-β expression in the clipped kidney, decreased proteinuria and restored protein plasma levels. In conclusion, transplantation into a renal subcapsule is an efficient route and MSC is a good candidate for cell therapy, which may represent an interesting approach for chronic kidney disease treatment. [ABSTRACT FROM AUTHOR]

Bone marrow cells (BMCs) are the main type of cells used for transplantation therapies. Obesity, a major world health problem, has been demonstrated to affect various tissues, including bone marrow. This could compromise the success of such therapies. One of the main mechanisms underlying the pathogenesis of obesity is mitochondrial dysfunction, and recent data have suggested an important role for mitochondrial metabolism in the regulation of stem cell proliferation and differentiation. Since the potential use of BMCs for clinical therapies depends on their viability and capacity to proliferate and/or differentiate properly, the analysis of mitochondrial function and cell viability could be important approaches for evaluating BMC quality in the context of obesity. We therefore compared BMCs from a control group (CG) and an obese group (OG) of mice and evaluated their mitochondrial function, proliferation capacity, apoptosis, and levels of proteins involved in energy metabolism. BMCs from OG had increased apoptosis and decreased proliferation rates compared with CG. Mitochondrial respiratory capacity, biogenesis, and the coupling between oxidative phosphorylation and ATP synthesis were significantly decreased in OG compared with CG, in correlation with increased levels of uncoupling protein 2 and reduced peroxisome proliferator-activated receptor-coactivator 1α content. OG also had decreased amounts of the glucose transporter GLUT-1 and insulin receptor (IRβ). Thus, Western-diet-induced obesity leads to mitochondrial dysfunction and reduced proliferative capacity in BMCs, changes that, in turn, might compromise the success of therapies utilizing these cells. [ABSTRACT FROM AUTHOR]

While Toxoplasma gondii is able to infect and replicate within all eukaryotic cells, tachyzoites are the infective form of T. gondii that invades all eukaryotic cells leading to tissue rupture, the main features of toxoplasmosis. The present study evaluates the activity of (benzaldehyde)-4-phenyl-3-thiosemicarbazone and (benzaldehyde)-(4 or 1)-phenylsemicarbazone against intracellular T. gondii. The nine new compounds were incubated in infected Vero cells at concentrations of 0.01, 0.1, 0.5, and 1.0 mM and evaluated for three main effects: cytotoxicity, infection, and number of intracellular parasites. The cytotoxicity test showed a pattern by analyzing the substituent arylhydrazone, where trihydroxy Compounds 4- 9 were cytotoxic at concentrations of 0.5 and 1.0 mM. The results highlight Compound 8, which reduced the number of intracellular parasites by 82 % in a concentration of 0.01 mM and showed a LD50 of 0.3 mM in cell culture. These different biological actions are due to changes in the molecular structure and type of radical present in each compound. All compounds tested were more efficient than the control drug sulfadizine. [ABSTRACT FROM AUTHOR]

Skeletal fibroblasts in vitro can acquire myofibroblast phenotypes by the development of biochemical and morphological features, mainly the expression of alpha-smooth-muscle actin (α-SMA). Myogenic differentiation is a central event in skeletal muscle development, and has commonly been studied in vitro in the context of skeletal muscle development and regeneration. Controlling this process is a complex set of interactions between myoblasts and the extracellular matrix. Osteopontin (OPN) is an acidic, phosphorylated matrix protein that contains an Arg-Gly-Asp (RGD) cell attachment sequence and has been identified as an adhesive and migratory substrate for several cell types. The aim of this study was to investigate osteopontin expression during the differentiation of skeletal fibroblasts into myofibroblasts and during myogenesis in a coculture model. Fibroblasts and myoblasts were obtained from skeletal muscle of 18-d-old Wistar strain rat fetuses by enzymatic dissociation. At 1 and 9 d, cocultures were immunolabeled, and the cells were also separately subjected to Western blotting to analyze OPN expression. Our data using confocal microscopy showed that myoblasts displayed a strong staining for OPN and that this labeling was maintained after myotube differentiation. Conversely, during fibroblast differentiation into myofibroblasts, we observed a significant increase in OPN expression. The results obtained by immunolabeling were confirmed by Western blotting. We suggest that OPN is important mainly during early stages of myogenesis, facilitating myoblast fusion and differentiation, and that the increased expression of OPN in myofibroblasts might be related to its effects as a key cytokine regulating tissue repair and inflammation. [ABSTRACT FROM AUTHOR]

The participation of resident, elicited, and activated macrophage surface components during internalization of tachyzoites of Toxoplasma gondii was analyzed using neuraminidase, phospholipase C, trypsin, protease, and hyaluronidase. Treatment of these macrophages with neuraminidase from Vibrio cholerae, phospholipase C from Bacillus cereus and Clostridium perfringens, protease, and hyaluronidase prior to their interaction with parasites increased the penetration of host cells by T. gondii. Incubation of macrophages with trypsin significantly inhibited the uptake of parasites. Our findings confirm previous observations that treatment of the macrophages with cytochalasin D under conditions that completely block the typical phagocytic process partially inhibits infection of the cells by T. gondii. The results of simultaneous treatment of the macrophages with enzymes and cytochalasin D suggested that the observed enhancement of cell infection by treatment with neuraminidase and hyaluronidase was attributable to a classic phagocytic process, whereas that obtained using phospholipase resulted from active penetration. [ABSTRACT FROM AUTHOR]

Fibroblasts in vitro can acquire myofibroblast phenotype by the development of several biochemical and morphological properties of smooth muscle cells, particularly the expresion of alpha-smooth muscle actin. These cells play a major role in inflammatory responses and in wound repair through their production of growth factors, cytokines, and other soluble mediators. Lipid bodies (LB) are lipid-rich cytoplasmic inclusions and have been recognized as specialized intracellular domains involved in the formation of paracrine mediators of inflammation. The aim of the present study was to investigate the occurrence and distribution of LB during differentiation of rat fetus skeletal fibroblasts into myofibroblasts in vitro. Primary cultures of fibroblasts were obtained from skeletal muscles of 18-d-old Wistar strain rat fetus by enzymatic dissociation. At 1–7 d, the cells were stained with Nile red vital dye to identify LB and then observed under a Zeiss CLSM-310. Our results showed that there was an accentuated increase in the number of LB during the differentiation of skeletal fibroblasts into myofibroblasts and that these inclusions were scattered at the cytoplasm. [ABSTRACT FROM AUTHOR]