Early proliferation of umbilical cord blood cells from premature neonates

BACKGROUND AND OBJECTIVE: Human umbilical cord blood (UCB) is an important source of haematopoietic stem cells; however, the behaviour of progenitor cells obtained from premature and full-term neonates is still a controversy subject. Thus, the aim of this study was to evaluate cell cycle parameters and the proliferative capacity of UCB progenitor cells from premature and full-term neonates. MATERIAL AND METHODS: Clonogenic assays were performed with methylcellulose, medium supplemented with recombinant stimulating growth factors and the colonies were scored on the seventh day and the 14th day of culture. A cell cycle study was carried out by DNA analysis using flow cytometry and 30 000 events were acquired; p107 and p130 expressions were analysed by Western blotting. RESULTS: Cultures obtained from UCB of premature neonates showed an early growth of colony-forming unit (CFU)-burst forming unit erythroid/CFU-granulocyte, erythrocyte, macrophage and megakaryocyte (BFU-E/GEMM), and CFU-granulocyte, macrophage (GM) by the seventh day of culture (P < 0.001). Therefore, the number and morphological characteristics of these colonies were comparable with those obtained from full-term neonates, on the 14th day of culture. At the 14th day, a large amount of CFU-GM was detected in the premature group (P < 0.0032). The premature culture on the 14th day showed fibroblasts and was comparable to those of full-term neonates on the 21st day in terms of number and morphology of the colonies. DNA analysis showed that the number of cells in S-phase was also higher in premature samples when compared to full-term neonates, P < 0.0021 (0 h = 12.8 vs. 2.5%; 16 h = 10.5 vs. 5.9%; 20 h = 13.5 vs. 10.3%; 24 h = 13.8 vs. 9.1%; 48 h = 14.0 vs. 5.4%; 72 h = 20.5 vs. 8.9%; 96 h = 13.8 vs. 7.7%). The Western blotting results demonstrated that p107 and p130 cell cycle protein expressions were higher in premature cells than in full-term cells. CONCLUSION: These results suggest that the higher capacity of proliferation and early differentiation of premature UCB might not be related only to the amount of stem/progenitor cells but also to a different timing of cell cycle entry.     

Administration of branched-chain amino acids alters epigenetic regulatory enzymes in an animal model of Maple Syrup Urine Disease

Maple Syrup Urine Disease (MSUD) is an autosomal recessive inherited disorder that affects the activity of the branched-chainα-keto acid dehydrogenase complex (BCDK). This deficiency on BCDK complex results in the accumulation of branched-chain amino acids (BCAA) leucine, isoleucine, valine, and their corresponding α-keto acids. Epigenetic changes can negatively affect the metabolism of BCAA. These changes are catalyzed by the epigenetic regulatory enzymes, e.g., DNA methyltransferase (DNMT), histone deacetylases (HDAC), and histone acetyltransferases (HAT). However, the impacts of BCAA administration on the activity of epigenetic regulatory enzymes in the brain of MSUD patients are still unknown. In this study, we aimed to demonstrate the impact of BCAA administration on the activity of DNMT, HDAC, and HAT in the brain structures of infant rats, an animal model of MSUD. For that, we administered a BCAA pool to infant rats for 21 days. We demonstrated that BCAA administration significantly increased the DNMT and HDAC activities in the hippocampus and striatum, but not in the cerebral cortex of MSUD infant rats. A positive correlation was observed between HDAC and DNMT activities in the hippocampus and striatum of animals exposed to BCAA injections. Our results showed that the BCAA administration could modulate epigenetic regulatory enzymes, mainly DNMT and HDAC, in the brains of infant rats. Therefore, we suggest that the increase in the activity of DNMT and HDAC in the hippocampus and striatum could partially explain the neurological impairments presented in animal models of MSUD.

     

Correlation between populations of Rhodnius and presence of palm trees as risk factors for the emergence of Chagas disease in Amazon region, Brazil

Among the states that comprise the legally defined Amazon region of Brazil, Pará has presented the highest occurrences of acute cases of Chagas disease over the last two decades. These cases have been attributed to consumption of fruits from native palm trees. In surveys in rural and wild areas of the municipality of Oriximiná, Pará, triatomine fauna, their main ecotopes and the infection rate due to Trypanosoma cruzi were identified using active and passive search methods: manual capture and Noireau traps, respectively. A total of 582 ecotopes were surveyed using 1496 Noireau traps. Out of 442 specimens collected, 289 were identified as Rhodnius robustus and 153 as Rhodnius pictipes. The infection rate caused by T. cruzi was 17.4%. The food sources of the triatomines were found to be birds, hemolymph, horses, and rodents. The association between R. robustus and inajá palm trees (Attalea marita), which are abundant in rural areas, was confirmed. On the other hand, R. pictipes is found in several palm tree species, such as inajá (A. marita), mucajá (Acrocomia aculeata), murumuru (Astrocaryum murumuru) and patauá (Oenocarpus bataua), and in bromeliads in wild areas. These occurrences of triatomine species in regions with or without T. cruzi infection, in the vicinity of the main settlement of the municipality, suggest that there is a need for entomological and epidemiological surveillance in this region.     

Titanium dioxide induced inflammation in the small intestine

AIM: To investigate the effects of titanium dioxide (TiO₂) nanoparticles (NPTiO₂) and microparticles (MPTiO₂) on the inflammatory response in the small intestine of mice.METHODS: Bl 57/6 male mice received distilled water suspensions containing TiO₂ (100 mg/kg body weight) as NPTiO₂ (66 nm), or MPTiO₂ (260 nm) by gavage for 10 d, once a day; the control group received only distilled water. At the end of the treatment the duodenum, jejunum and ileum were extracted for assessment of cytokines, inflammatory cells and titanium content. The cytokines interleukin (IL)-1b, IL-4, IL-6, IL-8, IL-10, IL-12, IL-13, IL-17, IL-23, tumor necrosis factor-α (TNF-α), intracellular interferon-γ (IFN-γ) and transforming growth factor-β (TGF-β) were evaluated by enzyme-linked immunosorbent assay in segments of jejunum and ileum (mucosa and underlying muscular tissue). CD4⁺ and CD8⁺ T cells, natural killer cells, and dendritic cells were evaluated in duodenum, jejunum and ileum samples fixed in 10% formalin by immunohistochemistry. The titanium content was determined by inductively coupled plasma atomic emission spectrometry.RESULTS: We found increased levels of T CD4⁺ cells (cells/mm²) in duodenum: NP 1240 ± 139.4, MP 1070 ± 154.7 vs 458 ± 50.39 (P < 0.01); jejunum: NP 908.4 ± 130.3, MP 813.8 ± 103.8 vs 526.6 ± 61.43 (P < 0.05); and ileum: NP 818.60 ± 123.0, MP 640.1 ± 32.75 vs 466.9 ± 22.4 (P < 0.05). In comparison to the control group, the groups receiving TiO₂ showed a statistically significant increase in the levels of the inflammatory cytokines IL-12, IL-4, IL-23, TNF-α, IFN-γ and TGF-β. The cytokine production was more pronounced in the ileum (mean ± SE): IL-12: NP 33.98 ± 11.76, MP 74.11 ± 25.65 vs 19.06 ± 3.92 (P < 0.05); IL-4: NP 17.36 ± 9.96, MP 22.94 ± 7.47 vs 2.19 ± 0.65 (P < 0.05); IL-23: NP 157.20 ± 75.80, MP 134.50 ± 38.31 vs 22.34 ± 5.81 (P < 0.05); TNFα: NP 3.71 ± 1.33, MP 5.44 ± 1.67 vs 0.99 ± 019 (P < 0.05); IFNγ: NP 15.85 ± 9.99, MP 34.08 ± 11.44 vs 2.81 ± 0.69 (P < 0.05); and TGF-β: NP 780.70 ± 318.50, MP 1409.00 ± 502.20 vs 205.50 ± 63.93 (P < 0.05).CONCLUSION: Our findings indicate that TiO₂ particles induce a Th1-mediated inflammatory response in the small bowel in mice.     

Saposins utilize two strategies for lipid transfer and CD1 antigen presentation

Transferring lipid antigens from membranes into CD1 antigen-presenting proteins represents a major molecular hurdle necessary for T-cell recognition. Saposins facilitate this process, but the mechanisms used are not well understood. We found that saposin B forms soluble saposin protein-lipid complexes detected by native gel electrophoresis that can directly load CD1 proteins. Because saposin B must bind lipids directly to function, we found it could not accommodate long acyl chain containing lipids. In contrast, saposin C facilitates CD1 lipid loading in a different way. It uses a stable, membrane-associated topology and was capable of loading lipid antigens without forming soluble saposin-lipid antigen complexes. These findings reveal how saposins use different strategies to facilitate transfer of structurally diverse lipid antigens.     

Abdominal actinomycosis

PURPOSE: Actinomyces israelii are normal inhabitants in the oral cavity and upper intestinal tract of humans. They rarely cause disease and are seldom reported as pathogens. As a pathogen it causes fistuli, sinuses, and may appear as an abdominal mass and/or abscess. The abdominal mass can masquerade as a malignant process that is very difficult to differentiate, often requiring surgical intervention with resection. Because of difficulty in making a preoperative diagnosis, we undertook this review to determine if all patients require surgical intervention and whether other adjunctive modalities may improve preoperative diagnosis. METHODS: We report two patients with abdominal actinomycosis, one affecting the sigmoid colon and the other the retroperitoneum, iliac crest region. Both simulated a malignant process and required operations for diagnosis and treatment. RESULTS: As reported, actinomycotic abdominal masses and strictures can be treated by penicillin alone. Long-term medical treatment seems to be very successful and avoids surgical resection. The difficulty is obtaining a definitive diagnosis. CONCLUSION: The computed tomographic scan is the most helpful diagnostic modality. Appearance of abdominal actinomycosis is usually a contrast enhancing multicystic lesion as was found in these two patients. Needle aspiration cytology can be fairly accurate in obtaining the diagnosis and is recommended for suspicious lesions.     

The human homologue of rat NG2, a chondroitin sulfate proteoglycan, is not expressed on the cell surface of normal hematopoietic cells but is expressed by acute myeloid leukemia blasts from poor-prognosis patients with abnormalities of chromosome band 11q23

In our efforts to produce monoclonal antibodies that recognize cell- surface antigens expressed by hematopoietic precursor and stromal cells, we generated a monoclonal antibody, 7.1, which recognizes a 220- to 240-kD cell-surface protein whose N-terminal amino acid sequence is identical to the rat NG2 chondroitin sulfate proteoglycan molecule. This chondroitin sulfate proteoglycan, previously reported to be expressed by human melanoma cells, was not found to be expressed by normal hematopoietic cells, nor was it expressed on the cell surface of cell lines of hematopoietic origin including cell lines with 11q23 abnormalities. It was found on the cell surface of acute myeloid leukemia (AML) blasts and cell lines derived from nonhematopoietic tissues. Samples of leukemic marrow from 166 children with AML enrolled on Childrens Cancer Group protocol 213 were evaluated for cell-surface expression of this proteoglycan molecule. In 18 of 166 (11%) patient samples, greater than 25% of leukemic blasts expressed the NG2 molecule. These 18 patients had a poorer outcome with respect to survival (P = .002) and event-free survival (P = .035) with an actuarial survival at 4 years of 16.7%. Blast cell expression of the NG2 molecule was strongly associated with French-American-British M5 morphology (P < .0001) and abnormalities in chromosome band 11q23, site of the MLL gene. These results show that the NG2 molecule is expressed by malignant hematopoietic cells that have abnormalities in chromosome band 11q23, suggesting that antibody 7.1 may be useful in the rapid identification of this group of poor-prognosis patients.     

Mandibular movements in children with and without signs and symptoms of temporomandibular disorders

This research aimed to evaluate mandibular movements in children with and without signs and symptoms of temporomandibular dysfunction. The sample taken consisted of 99 children aged 3 to 5 years distributed in two groups: I - Absence of signs and/or symptoms of TMD (25 girls/40 boys); II - Presence of signs and symptoms of TMD (16 girls/18 boys). The symptoms were evaluated through an anamnesis questionnaire answered by the child's parents/caretakers. The clinical signs were evaluated through intra- and extraoral examination. Maximum mouth opening and left/right lateral movements were measured using a digital caliper. The maximum protrusive movement was measured using a millimeter ruler. The means and standard deviations for maximum mouth opening in Group I and Group II were 40.82mm±4.18 and 40.46mm±6.66, respectively. The values found for the left lateral movement were 6.96mm±1.66 for Group I and 6.74mm±1.55 for Group II, while for the right lateral movement they were 6.46mm±1.53 and 6.74mm±1.77. The maximum protrusion movements were 5.67mm±1.76 and 6.12mm±1.92, in Groups I and II, respectively. The mandibular movement ranges neither differed statistically between groups nor between genders. FAPESP Process 96/0714-6.     

The influence of a novel propolis on mutans streptococci biofilms and caries development in rats

A flavonoids-free Brazilian propolis (type 6) showed biological effects against mutans streptococci and inhibited the activity of glucosyltransferases. This study evaluated the influence of the ethanolic extract of a novel type of propolis (EEP) and its purified hexane fraction (EEH) on mutans streptococci biofilms and the development of dental caries in rats. The chemical composition of the propolis extracts were examined by gas chromatography/mass spectrometry. The effects of EEP and EEH on Streptococcus mutans UA159 and Streptococcus sobrinus 6715 biofilms were analysed by time-kill and glycolytic pH drop assays. Their influence on proton-translocating F-ATPase activity was also tested. In the animal study, the rats were infected with S. sobrinus 6715 and fed with cariogenic diet 2000. The rats were treated topically twice a day with each of the extracts (or control) for 5 weeks. After the experimental period, the microbial composition of their dental plaque and their caries scores were determined. The results showed that fatty acids (oleic, palmitic, linoleic and stearic) were the main compounds identified in EEP and EEH. These extracts did not show major effects on the viability of mutans streptococci biofilms. However, EEP and EEH significantly reduced acid production by the biofilms and also inhibited the activity of F-ATPase (60-65%). Furthermore, both extracts significantly reduced the incidence of smooth surface caries in vivo without displaying a reduction of the percentage of S. sobriuns in the animals' plaque (P < 0.05). However, only EEH was able to reduce the incidence and severity of sulcal surface caries (P < 0.05). The data suggest that the cariostatic properties of propolis type 6 are related to its effect on acid production and acid tolerance of cariogenic streptococci; the biological activities may be attributed to its high content of fatty acids.     

Evaluation of marginal microleakage in class II cavities: effect of microhybrid, flowable, and compactable resins.

OBJECTIVE: The goal of the present study was to evaluate the microleakage on the cementum/dentin and enamel surfaces in Class II restorations, using different kinds of resin composite (microhybrid, flowable, and compactable). METHOD AND MATERIALS: Forty human caries-free molars were extracted and selected. Eighty Class II standardized cavities were made in the cervical wall at the cementoenamel junction (CEJ) and at the mesial and distal surfaces. The teeth were divided into four groups: G1--adhesive system + microhybrid resin composite Z100; G2--adhesive system + compactable resin composite Prodigy Condensable; G3--adhesive system + flowable resin composite Revolution + Z100 resin composite; G4--adhesive system + Revolution fluid resin + compactable resin composite Prodigy Condensable. The adhesive system used in this study was Scotchbond Multi-Purpose Plus. The specimens were thermocycled in baths of 5 degrees C and 55 degrees C for 1,000 cycles and immersed in 50% silver nitrate solution. The specimens then were sectioned and evaluated on degree of dye penetration. RESULTS: The results were evaluated using the nonparametric Kruskall-Wallis test, which showed a statistically significant difference between groups G1 and G4, G2 and G4, and G3 and G4. CONCLUSIONS: None of the materials was able to eliminate the marginal microleakage at the cervical wall; the application of a low-viscosity resin composite combined with a compactable resin composite significantly decreased the microleakage.