Radiation-induced Hierarchical Topologically Associated Domain Change and Its Role in Cellular Responses to Radiation / 中国生物化学与分子生物学报

The three-dimensional (3D) genome organization plays an important role in gene regulation. As a basic functional unit of the genome, topologically associated domain (TAD) regulates multiplebiological processes such as gene expression and DNA replication and plays a role in radiation-inducedDNA damage repair. Recent studies showed that TAD is not a completely independent domain butcontains hierarchical internal domains, which could be a new mechanism of gene regulation. To explorethe role of hierarchical TAD in cellular responses to radiation, we apply the OnTAD algorithm, anoptimized nested TAD caller from Hi-C data, to identify hierarchical TAD in 26 Hi-C data from Geneexpression omnibus (GEO) database. These data were from irradiated fibroblasts, lymphoblasts andfibroblasts deficient in the ataxia telangiectasia mutated (ATM) gene with 5 Gy X-ray. We observe thatX-ray can regularly affect the hierarchy of TAD in which high-level TAD is prone to change and low-levelTAD is more conservative. We propose that radiation-induced TAD hierarchy change can regulate cellularresponses to radiation by regulating gene expression, and ATM is a necessary factor for radiation-inducedTAD hierarchy change and recovery. This study provides new insights into the role of the 3D genome inradiation-induced cellular responses from the perspective of hierarchical TAD structures.

A literature review on implant abutment and soft tissue response

In the implant prosthetic procedure, the soft tissue reaction was varied with the material and surface treatment of the abutment. It may be the cause of the peri-implantitis, and hence it can affect the long-term prognosis of the implant prosthesis. Titania and zirconia abutment presented superior biocompatibility and stable soft tissue reaction, while gold alloy abutment showed unfavorable reaction sometimes. A soft tissue reaction can be differed by the surface characteristics even in the same material type. Because rougher surface induces a bacterial attachment, the part contacting a soft tissue should have smooth surface. Additional surface treatment can enhance the cellular response without increasing bacterial attachment. Repeated removal and insertion of the abutment and the shape of the abutment may affect the soft tissue reaction, also. Ultrasonic cleaning and argon plasma cleaning are effective way to clean the retained micro-dust on the customized abutment.

Respuesta in vitro de células mononucleares de sangre periférica de mujeres embarazadas infectadas con Toxoplasma gondii / In vitro Response of of peripheral blood mononuclear cells of pregnant women infected with Toxoplasma gondii

Objetivo: evaluar in vitro la respuesta inmune de células mononucleares de sangre periférica de mujeres embarazadas infectadas con Toxoplasma gondii. Metodología: los grupos de estudio fueron mujeres embarazadas con toxoplasmosis (E+T+) (n=15), mujeres embarazadas sin toxoplasmosis (E+T-) (n=15) y mujeres no embarazadas sin toxoplasmosis (E-T-) (n=15). células mononucleares de sangre periférica (PBMC) de los tres grupos de estudio, fueron estimuladas in vitro, con antígeno soluble de T.gondii. Transcurrido el tiempo de incubación (6 días), se midió los niveles de proliferación celular y producción de IFN-y e IL-10 (citoquinas TH1 y TH2 respectivamente) este último en sobrenadantes de cultivo, adicionalmente se realizó el recuento absoluto de linfocitos T CD3+, CD4+, CD8+, en muestras de sangre total tratadas con EDTA. Resultados: los resultados muestran una mayor proliferación celular en el grupo (E+T+) diferencias estadísticamente significativas en comparación con los grupos (E+T-) y (E-T-). Respecto a los niveles de citoquinas TH1 se observó mayor producción de INF-y en el grupo (E+T+), comparado con los grupos (E+T-) y (E-T-). Producción de IL-10 son bajos y/o no detectables en los tres grupos de estudio. Los niveles de CD3+, CD4+, CD8+ se encontraron dentro los parámetros normales en los tres grupos de estudio, sin embargo en la relación CD4+/CD8+, en los grupos (E+T-) y (E-T-) presentan valores disminuidos en comparación al grupo (E-T-). Conclusiones: en mujeres embarazadas con toxoplasmosis (E+T+) existe una polarización de la respuesta inmune haciaTHl, brindándoles de alguna manera inmunidad frente a una posible reinyección con el parásito, a diferencia de los grupos (E+T-) y (E-T-) que podrían ser más susceptibles de contraer toxoplasmosis durante el embarazo, ya que sus niveles de IFN-y e IL-10 son bajos y/o no detectables. Respecto a los niveles de linfocitos T CD3+, CD4+, CD8+ de las mujeres embarazadas, si bien están dentro los parámetros normales, se encuentran levemente disminuidos, esto podría deberse al estado inmunogulador que presentan las mujeres durante el embarazo, sin embargo se podría decir que presentan protección contra T. gondii.

Objective: To evaluate in vitro the immune response of peripheral blood mononuclear cells of pregnant women infected with Toxoplasma gondii. Methodology: The study groups were pregnant women with toxoplasmosis (E + T +) (n = 15), pregnant women without toxoplasmosis (E + T) (n = 15) and non-pregnant women without toxoplasmosis (ET) (n = fifteen). Peripheral blood mononuclear cells (PBMC) of the three study groups were stimulated in vitro with T. gondii soluble antigen. After the incubation period (6 days), levels of cell proliferation and IFN-y and IL-10 (TH1 and TH2 respectively) this last in culture supernatants, additionally it was performed the absolute count of T CD3 +, CD4 +, CD8 + lymphocyte, in whole blood samples treated with EDTA. Results: The results show increased cell proliferation in the group (E + T +) statistically significant differences compared to the groups (E + T) and (ET). Regarding TH1 cytokine levels, it was observed higher production of IFN-y in group (E + G +) compared with groups (E + T) and (ET-). Production of IL-10 is low and / or undetectable in all three study groups. The levels of CD3 +, CD4 +, CD8 + were within normal parameters in the three study groups, but in the CD4 + / CD8 + in the groups (E + T) and (ET) have decreased compared values to group (ET). Conclusions: Pregnant women infected with Toxoplasmosis (E + T +) there is a polarization of the immune response haciaTH1, so providing some immunity against a possible reinjection with the parasite, as opposed to groups (E + T) and (ET- ) that may be more susceptible to toxoplasmosis during pregnancy because their levels of IFN-y and IL-10 are low and / or undetectable. Regarding the levels of CD3 + T CD4 +, CD8 + lymphocytes of pregnant women, if they are well within normal parameters, are slightly decreased, this could be due to inmunogulador been presented by women during pregnancy, but you could say that they have protection against T. gondii.

Characteristics and osteogenic effect of zirconia porous scaffold coated with beta-TCP/HA

PURPOSE: The purpose of this study was to evaluate the properties of a porous zirconia scaffold coated with bioactive materials and compare the in vitro cellular behavior of MC3T3-E1 preosteoblastic cells to titanium and zirconia disks and porous zirconia scaffolds. MATERIALS AND METHODS: Titanium and zirconia disks were prepared. A porous zirconia scaffold was fabricated with an open cell polyurethane disk foam template. The porous zirconia scaffolds were coated with beta-TCP, HA and a compound of beta-TCP and HA (BCP). The characteristics of the specimens were evaluated using scanning electron microscopy (SEM), energy dispersive x-ray spectrometer (EDX), and x-ray diffractometry (XRD). The dissolution tests were analyzed by an inductively coupled plasma spectrometer (ICP). The osteogenic effect of MC3T3-E1 cells was assessed via cell counting and reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: The EDX profiles showed the substrate of zirconia, which was surrounded by the Ca-P layer. In the dissolution test, dissolved Ca2+ ions were observed in the following decreasing order; beta-TCP > BCP > HA (P<.05). In the cellular experiments, the cell proliferation on titanium disks appeared significantly lower in comparison to the other groups after 5 days (P<.05). The zirconia scaffolds had greater values than the zirconia disks (P<.05). The mRNA level of osteocalcin was highest on the non-coated zirconia scaffolds after 7 days. CONCLUSION: Zirconia had greater osteoblast cell activity than titanium. The interconnecting pores of the zirconia scaffolds showed enhanced proliferation and cell differentiation. The activity of osteoblast was more affected by microstructure than by coating materials.

The effect of Ca-P coatings of anodized implant surface on response of osteoblast-like cells in vitro / 대한치과보철학회지

PURPOSE: The purpose of this study was to evaluate the response of osteoblast-like cells to Ca-P coated surface obtained via Ion beam-assisted deposition (IBAD) method and Sol-Gel process on anodized surface by cellular proliferation and differentiation. MATERIAL AND METHODS: The surface of a commercially pure titanium (Grade IV) discs with dimension of 10mm diameter and 2 mm thickness was modified by anodic oxidation under a constant voltage of 300 V. The experimental groups were coated with Ca-P by the IBAD method and Sol-Gel process on anodized surface. The surface roughness (Ra) of specimens was measured by optical interferometer and each surface was examined by SEM. To evaluate cell response, MG63 cells were cultured and cell proliferation, ALP activity and the ability of cell differentiation were examined. Also, cell morphology was examined by SEM. The significant of each group was verified by Kruskal-Wallis Test (alpha= .05). RESULTS: The Ra value of Ca-P coated surface by IBAD method was significantly higher than Ca-P coated surface by Sol-gel process (P<.05). The level of cell proliferation and ALP activity was higher in Ca-P coated surface by IBAD method (P<.05). The expression of ALP showed higher level expression in Ca-P coated surface by IBAD method. Cells grown on Ca-P coated surface by IBAD method were uniformly distributed and developed a very close layer. CONCLUSION: These experiments showed better performances of Ca-P coated surface by IBAD method with respect to Ca-P coated surface by Sol-gel process. Ca-P coated surface by IBAD method appear to give rise more mature osteoblast characteristics and might result in increased bone growth and bone-implant contact.

Vacina de DNA: aspectos gerais e sua aplicação na medicina humana e veterinária / DNA vaccines: general concerns and its applications in human and veterinary medicine

A vacinação com DNA é uma das mais promissoras técnicas de imunização contra uma variedade de patógenos e tumores, para os quais os métodos convencionais não tem sido eficientes. Vacinas de DNA são capazes de induzir resposta imune humoral e celular, tanto para resposta de linfócitos CD4 + quanto CD8 +, sem a necessidade de microrganismos vivos. Apesar do grande potencial de induzir imunidade protetora, a vacina de DNA nem sempre apresenta bons resultados. A imunidade depende de vários fatores como a seleção do gene alvo, construção do vetor de expressão, freqüência e via de administração da vacina, quantidade de DNA, localização do antígeno codificado pelo plasmídio e idade, saúde e espécies de animais vacinados. Esta revisão relata o desenvolvimento de algumas vacinas de DNA para doenças de interesse na medicina veterinária e humana.

The vaccination with DNA is one of the most promising immunization techniques against a pathogensvariety and tumors, for which the conventional methods have not been efficient. DNA vaccines arecapable to induce immune humoral and cellular response, directed to lymphocytes CD4+ and CD8+,without the necessity of live microorganisms. In spite of the great potential of inducing protectiveimmunity, the DNA vaccine not always has success. The immunity depends on several factors such asthe selection of the target gene, construction of the expression vector, frequency and via of administrationof the vaccine, amount of DNA, location of the antigen codified by the plasmid and age, health andspecies of vaccinated animals. This revision shows the development of some vaccines of DNA fordiseases of interest in the veterinary and human medicine.

Surface characteristics and biological responses of hydroxyapatite coating on titanium by hydrothermal method: an in vitro study / 대한치과보철학회지

STATEMENT OF PROBLEM: Hydroxyapatite(HA) coated titanium surfaces have not yet showed the reliable osseointegration in various conditions. PURPOSE: This study was aimed to investigate microstructures, chemical composition, and surface roughness of the surface coated by the hydrothermal method and to evaluate the effect of hydrothermal coating on the cell attachment, as well as cell proliferation. MATERIAL AND METHODS: Commercially pure(c.p.) titanium discs were used as substrates. The HA coating on c.p. titanium discs by hydrothermal method was performed in 0.12M HCl solution mixed with HA(group I) and 0.1M NaOH solution mixed with HA(group II). GroupI was heated at 180degrees C for 24, 48, and 72 hours. GroupII was heated at 180degrees C for 12, 24, and 36 hours. And the treated surfaces were evaluated by Scanning electron microscopy(SEM), Energy dispersive X-rayspectroscopy(EDS), X-ray photoelectron spectroscopy(XPS), X-ray diffraction method(XRD), Confocal laser scanning microscopy(CLSM). And SEM of fibroblast and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) assay were used for cellular responses of the treated surfaces. RESULTS: The color of surface changed in both groups after the hydrothermal process. SEM images showed that coating pattern was homogeneous in group II, while inhomogeneous in group I. H72 had rosette-like precipitates. The crystalline structure grew gradually in group II, according to extending treatment period. The long needle-like crystals were prominent in N36. Calcium(Ca) and phosphorus(P) were not detected in H24 and H48 in EDS. In all specimens of group II and H72, Ca was found. Ca and P were identified in all treated groups through the analysis of XPS, but they were amorphous. Surface roughness did not increase in both groups after hydrothermal treatment. The values of surface roughness were not significantly different between groups I and II. According to the SEM images of fibroblasts, cell attachments were oriented and spread well in both treated groups, while they were not in the control group. However, no substantial amount of difference was found between groups I and II. CONCLUSIONS: In this study during the hydrothermal process procedure, coating characteristics, including the HA precipitates, crystal growth, and crystalline phases, were more satisfactory in NaOH treated group than in HCl treated group. Still, the biological responses of the modified surface by this method were not fully understood for the two tested groups did not differ significantly. Therefore, more continuous research on the relationship between the surface features and cellular responses seems to be in need.

A study on osteoblast-like cell responses to surface-modified titanium / 대한치과보철학회지

STATEMENT OF PROBLEM: The success of implants depends on intimate and direct contact of implant material on bone tissue and on functional relationship with soft tissue contact. Creation and maintenance of osseointegration depend on the understanding of the tissue's healing, repairing, and remodeling capacity and these capacities rely on cellular behavior. Altering the surface properties can modify cellular responses such as cell adhesion, cell motility, bone deposition. Therefore, various implant surface treatment methods are being developed for the improved bone cell responses. PURPOSE: The purpose of this study was to evaluate the responses of osteoblast-like cells to surface- modified titanium. MATERIALS AND METHODS: The experiment was composed of four groups. Group 1 represented the electropolished surface. Group 2 surfaces were machined surface. Group 3 and Group 4 were anodized surfaces. Group 3 had low roughness and Group 4 had high roughness. Physicochemical properties and microstructures of the d iscs were examined and the responses of osteoblast-like cells to the discs were investigated. The microtopography was observed by SEM. The roughness was measured by three-dimension roughness measuring system. The microstructure was analyzed by XRD, AES. To evaluate cell responses to modified titanium surfaces, osteoblasts isolated from calvaria of neonatal rat were cultured. Cell count, morphology, total protein measurement and alkaline phosphatase activities of the cultures were examined. RESULTS AND CONCLUSION: The results were as follows 1. The four groups showed specific microtopography respectively. Anodized group showed grain structure with micropores. 2. Surface roughness values were, from the lowest to the highest, electropolished group, machined group, low roughness anodized group, and high roughness anodized group. 3. Highly roughened anodized group was found to have increased surface oxide thickness and surface crystallinity. 4. The morphology of cells, flattened or spherical, were different from ach other. In the electropolished group and machined group, the cells were almost flattened. In two anodized groups, some cells were spherical and other cells were flattened. And the 14 day culture cells of all of the groups were nearly flattened due to confluency. 5. The number of attached cells was highest in low roughness anodized group. And the machined group had significantly lower cell count than any other groups(P<.05). 6. Total protein contents showed no difference among groups. 7. The level of alkaline phosphatase activities was higher in the anodized groups than electropolished and machined groups(P<.05).