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1.
Am J Trop Med Hyg ; 101(1): 207-213, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31162004

RESUMO

rVSVΔG-ZEBOV-GP vaccine is a live recombinant (r) vesicular stomatitis virus (VSV), where the VSV G protein is replaced with the Zaire Ebola virus (ZEBOV) glycoprotein (GP). For vaccine immunogenicity testing, clinical trial sera collected during an active ZEBOV outbreak underwent gamma irradiation (GI) before testing in biosafety level 2 laboratories to inactivate possible wild-type ZEBOV. Before irradiating pivotal trial samples, two independent studies evaluated the impact of GI (50 kGy) on binding ZEBOV-GP (ELISA) antibodies against rVSVΔG-ZEBOV-GP, using sera from a North American phase 1 study. Gamma irradiation was associated with slightly higher antibody concentrations in pre-vaccination samples and slightly lower concentrations postvaccination. Results indicate that GI is a viable method for treating samples from regions where filoviruses are endemic, with minor effects on antibody titers. The impact of GI on immunogenicity analyses should be considered when interpreting data from irradiated specimens.


Assuntos
Anticorpos Antivirais/efeitos da radiação , Vacinas contra Ebola/imunologia , Ebolavirus/metabolismo , Raios gama , Soro/efeitos da radiação , Anticorpos Antivirais/sangue , Anticorpos Antivirais/fisiologia , Doença pelo Vírus Ebola/epidemiologia , Doença pelo Vírus Ebola/imunologia , Doença pelo Vírus Ebola/prevenção & controle , Humanos , Glicoproteínas de Membrana , Vacinação , Vacinas Sintéticas/imunologia , Proteínas do Envelope Viral/imunologia
2.
Molecules ; 23(5)2018 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-29701645

RESUMO

The diamondback moth, Plutella xylostella (Linnaeus), is one of the notorious pests causing substantial loses to many cruciferous vegetables across the nations. The effects of 60Co-γ radiation on physiology of P. xylostella were investigated and the results displayed that 200 Gy irradiation significantly alters the antioxidant enzyme regulation in six-day-old male pupae of P. xylostella. First, in our research, we detected Oxidase system and stress response mechanism of irradiated pupae, the results displayed that 200 Gy irradiation significantly alters the antioxidant enzyme regulation in six-day-old male pupae of P. xylostella. The levels of superoxide dismutase (SOD) and catalase (CAT) were increased significantly in contrast the level of peroxidase (POD) and glutathione S-transferase (GST) were decreased in 12⁻24 h post-treatment. The heat shock proteins (Hsps) gene expression level was significant increasing, maximum > 2-folds upregulation of genes were observed in peak. However, they also had a trend of gradual recovery with development. Second, we detected the testis lactate dehydrogenase (LDH) and acid phosphatase (ACP) activity found that in male adults testis they increased significantly than control during its development. Thus the present research investigation highlights that the 60Co-γ radiation treatments alters the physiological development of diamondback moth. The results showed that 200 Gy dosage resulted in stress damage to the body and reproductive system of the diamondback moth.


Assuntos
Antioxidantes/metabolismo , Radioisótopos de Cobalto/efeitos adversos , Proteínas de Choque Térmico HSP70/genética , Lepidópteros/efeitos da radiação , Soro/química , Fosfatase Ácida/metabolismo , Animais , Antioxidantes/efeitos da radiação , Raios gama/efeitos adversos , Regulação da Expressão Gênica/efeitos da radiação , Proteínas de Insetos/genética , L-Lactato Desidrogenase/metabolismo , Lepidópteros/enzimologia , Lepidópteros/metabolismo , Camundongos , Soro/efeitos da radiação
3.
Molecules ; 23(5)2018 04 26.
Artigo em Inglês | MEDLINE | ID: mdl-29701672

RESUMO

Chemometric analysis of bioactive compounds revealed that American ginsengs (AGs) from different cultivation regions of China had a difference in quality, which indicates their possible pharmacological difference. A UPLC-Q/TOF-MS-based untargeted metabolomic approach was used to uncover serum metabolite changes in radiated mice pre-administered with AG root decoctions from seven cultivation regions and to further assess their quality difference. OPLS-DA revealed that 51 metabolites (ESI−) and 110 (ESI⁺) were differentially expressed in sera between the control and the radiated model mice. Heatmap analysis further revealed that AG could not reverse most of these radiation-altered metabolites, which indicates dietary supplement of AG before cobalt radiation had the weak potential to mediate serum metabolites that were altered by the sub-lethal high dose radiation. In addition, 83 (ESI−) and 244 (ESI⁺) AG altered metabolites were detected in radiated mice under radiation exposure. Both OPLS-DA on serum metabolomes and heatmap analysis on discriminant metabolites showed that AGs from different cultivation regions differentially influenced metabolic alterations in radiated mice, which indicates AGs from different cultivation regions showed the pharmacological difference in modulation of metabolite changes. AGs from Shandong, Shanxi, and Beijing provinces had more similar pharmacological effects than AGs from USA, Canada, Jilin, and Heilongjiang. Finally, 28 important potential biomarkers were annotated and assigned onto three metabolic pathways including lipid, amino acid, and energy metabolisms.


Assuntos
Radioisótopos de Cobalto/efeitos adversos , Medicamentos de Ervas Chinesas/administração & dosagem , Metabolômica/métodos , Panax/química , Soro/química , Animais , Cromatografia Líquida de Alta Pressão , Espectrometria de Massas , Metaboloma/efeitos dos fármacos , Metaboloma/efeitos da radiação , Camundongos , Panax/classificação , Raízes de Plantas/química , Soro/efeitos dos fármacos , Soro/efeitos da radiação
4.
Acta Biochim Pol ; 64(1): 189-193, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-27815965

RESUMO

Radiotherapy causes molecular changes observed at the level of body fluids, which are potential biomarker candidates for assessment of radiation exposure. Here we analyzed radiotherapy-induced changes in a profile of small metabolites detected in sera of head and neck cancer patients using the gas chromatography coupled with mass spectrometry approach. There were about 20 compounds, including carboxylic acids, sugars, amines and amino acids, whose levels significantly differed between pre-treatment and post-treatment samples. Among metabolites upregulated by radiotherapy there was 3-hydroxybutyric acid, whose level increased about three times in post-treatment samples. Moreover, compounds affected by irradiation were associated with several metabolic pathways, including protein biosynthesis and amino acid metabolism.


Assuntos
Ácido 3-Hidroxibutírico/metabolismo , Carcinoma de Células Escamosas/radioterapia , Neoplasias de Cabeça e Pescoço/radioterapia , Radiação Ionizante , Soro/metabolismo , Ácido 3-Hidroxibutírico/sangue , Ácido 3-Hidroxibutírico/efeitos da radiação , Adulto , Idoso , Biomarcadores , Carcinoma de Células Escamosas/metabolismo , Feminino , Cromatografia Gasosa-Espectrometria de Massas , Neoplasias de Cabeça e Pescoço/metabolismo , Humanos , Masculino , Redes e Vias Metabólicas/efeitos da radiação , Metabolômica/métodos , Pessoa de Meia-Idade , Exposição à Radiação/análise , Soro/efeitos da radiação , Regulação para Cima/efeitos da radiação
5.
Pak J Pharm Sci ; 29(1 Suppl): 351-5, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27005501

RESUMO

The blood serum rheological properties open the door to find suitable radio-protectors and convenient therapy for many cases of radiation exposure. The present study aimed to investigate the rheological properties of rat blood serum at wide range of shear rates after whole body irradiation with different gamma radiation doses in vivo. Healthy male rats were divided into five groups; one control group and 4 irradiated groups. The irradiation process was carried out using Co60 source with dose rate of 0.883cG/sec. Several rheological parameters were measured using Brookfield LVDV-III Programmable rheometer. A significant increase in viscosity and shear stress was observed with 25 and 50Gy corresponding to each shear rate compared with the control; while a significant decrease observed with 75 and 100Gy. The viscosity exhibited a Non-Newtonian behaviour with the shear rate while shear stress values were linearly related with shear rate. The decrease in blood viscosity might be attributed to changes in molecular weight, pH sensitivity and protein structure. The changes in rheological properties of irradiated rats' blood serum might be attributed to destruction changes in the haematological and dimensional properties of rats' blood products.


Assuntos
Raios gama , Hemorreologia , Soro/química , Soro/efeitos da radiação , Animais , Proteínas Sanguíneas/química , Viscosidade Sanguínea/efeitos da radiação , Concentração de Íons de Hidrogênio , Masculino , Peso Molecular , Dosagem Radioterapêutica , Ratos , Ratos Endogâmicos WKY , Resistência ao Cisalhamento/efeitos da radiação
6.
J Virol Methods ; 225: 16-22, 2015 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-26335960

RESUMO

Canine serum preserved at room temperature (25°C) for longer than 24h is known to exhibit significant cytotoxicity. This phenomenon is one of the major reasons for the failure of virus neutralization tests. In this study, a method for reducing this cytotoxicity was investigated by applying several treatments to dog, cat and human serum prior to room temperature storage. Additionally, the identity of the cytotoxic factor generated during room temperature storage was investigated. Heat-inactivation at 56°C or 65°C and the addition of protease inhibitor prior to storage were found to be effective for reducing cytotoxicity in the serum. Furthermore, heat-inactivation at 65°C reduced the cytotoxicity that was induced under room temperature storage. Several protein factors in serum were suspected to play a role in the observed cytotoxicity. According to this study, the membrane-attack-complex in serum was not involved in the cytotoxicity. This study provides useful information for development and improvement of cell culture and virus neutralization tests.


Assuntos
Proteínas Sanguíneas/toxicidade , Técnicas de Cultura de Células/métodos , Soro/química , Manejo de Espécimes/métodos , Animais , Gatos , Cães , Humanos , Soro/efeitos da radiação , Temperatura , Fatores de Tempo
7.
Rev. esp. nutr. comunitaria ; 20(supl.1): 20-25, mar. 2014. tab, graf
Artigo em Inglês | IBECS | ID: ibc-128006

RESUMO

Background: Good hydration status (HS) is necessary for an adequate homeostasis of the organism. Cytokines are secreted mainly by inflammatory leukocytes and act as intercellular mediators. Objetive: Assessing pro and anti-inflammatory cytokines concentration in serum and in the aqueous phase of stools (APhS) from healthy adults in function of their HS. Methods: HS data were obtained from 86 healthy adults of 45-65 years old and BMI ≥18.5-<40 kg/m2. HS was measured by bioelectrical impedance (BIA) with a standardized protocol. Cytokines serum concentrations were determined by multiple ELISAs. Stools were recollected by the participants, frozen, and carefully transported to the laboratory where they were stored at -80°C until their determination. Stools were ultra-centrifuged and cytokines were measured in APhS with an ultra-sensible cytokines array. All samples were analyzed in duplicate. Results: Mean age was 51.2 ± 4.9 years old and BMI was 28.2 ± 4.7 kg/m2. The average intake of water from foods and beverages was not adequate enough (1,411.6 ± 427.4 ml/day; 81% consumed less than two-thirds of the recommended intake) however only 89.5% showed an adequate HS and only 10.5% showed clearly dehydration measured by BIA. Volunteers who had good HS had lower values of IFN(2.7 ± 2.4 vs 6.4 ± 4.3 pg/ml; p < 0.05) and IL6 serum (5.5 ± 13.3 vs 6.4 ± 16.3 pg/ml; p < 0.01) than those who had a dehydration status. IL1 from AphS showed lower values in adults with good hydration than those dehydrated (648.3 ± 615 vs 1,194 ± 561.2 pg/ml; p < 0.05). Conclusions: Adults with an appropriate HS have a minor concentration of pro-inflammatory cytokines in serum and in APhS than adults who showed a dehydration status. More studies are needed in order to corroborate these results (AU)


Introducción: Un adecuado estado de hidratación (EH) es necesario para mantener la homesostasis del organismo. Las citoquinas son mediadores intercelulares que son secretadas principalmente por leucocitos. Objetivo: Valorar la concentración de citoquinas pro y antiinflamatorias en suero y la fase acuosa de las heces (FAH) de adultos sanos en función de su EH. Métodos: Se obtuvo información sobre el EH de 86 adultos sanos de 45-65 años y un IMC de 18,5-<40 kg/m2. El EH fue medido por Impedancia Bioeléctrica (BIA) siguiendo el protocolo estándar. La concentración de citoquinas en suero fue determinada por múltiples ELISAs. Las heces fueron recolectadas por los participantes, congeladas y transportadas al laboratorio donde fueron almacenadas a -80°C hasta su determinación. Posteriormente las heces fueron ultracentrifugadas y las citoquinas fueron medidas en la FAH con un array ultrasensible. Resultados: La edad media fue de 51,2 ± 4,9 años y el IMC fue de 28,2 ± 4,7 kg/m2. El consumo medio de agua proveniente de los alimentos y las bebidas realizado por los participantes no fué suficiente (1.411,6 ± 427,4 ml/día; el 81% consumió menos de dos tercios de la ingesta recomendada), sin embargo, el 89,5% presentó un adecuado EH y solo el 10,5% estuvo en rango de deshidratación. Los participantes con un adecuado EH tuvieron valores de IFN(2,7 ± 2,4 vs 6,4 ± 4,3 pg/ml; p < 0,05) e IL6 séricos (5,5 ± 13,3 vs 6,4 ± 16,3 pg/ml; p < 0,01) inferiores a las personas deshidratadas. La IL1 medida en la FAH mostró una concentración más baja en personas bien hidratadas que en aquellas deshidratadas (648,3 ± 615 vs 1194,0 ± 561,2 pg/ml; p < 0,05). Conclusiones: Los adultos de nuestro estudio con un adecuado EH presentan una concentración inferior de citoquinas proinflamatorias en suero y en la FAH que aquellos que estaban deshidratados. Se necesitan más estudios que confirmen estos resultados (AU)


Assuntos
Humanos , Masculino , Feminino , Adulto , Voluntários Saudáveis/estatística & dados numéricos , Homeostase/fisiologia , Citocinas/metabolismo , Citocinas/fisiologia , Soro/química , Soro/fisiologia , Soro/efeitos da radiação , Fezes/química , Fezes/citologia , Impedância Elétrica , Desidratação/dietoterapia , Desidratação/diagnóstico
8.
Adv Healthc Mater ; 3(4): 536-42, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24002989

RESUMO

The non-destructive ex vivo determination of haemoglobin (Hgb) concentration offers the capability to conduct multiple red blood cell haematological measurements on a single sample, an advantage that current optical techniques are unable to offer. Here, a microwave method and device for the accurate and non-destructive determination of Hgb concentration in microlitre blood samples are described. Using broadband microwave spectroscopy, a relationship is established between the dielectric properties of murine blood and Hgb concentration that is utilized to create a technique for the determination of Hgb concentration. Subsequently, a microwave dielectric resonator-microfluidic system is implemented in the analysis of 52 murine samples with microlitre volumes and Hgb concentrations ranging from 0 to 17 g dL(-1) . Using the characterized relationship, independent and minimally invasive Hgb measurements are made on nine healthy mice as well as seven with mutations in the Adenomatous polyposis coli (APC) gene that leads to colorectal cancer and consequently anaemia.


Assuntos
Eritrócitos/química , Hemoglobinas/análise , Micro-Ondas , Animais , Proteínas Sanguíneas/análise , Proteínas Sanguíneas/química , Espectroscopia Dielétrica , Eletrólitos/sangue , Eletrólitos/química , Eritrócitos/efeitos da radiação , Camundongos , Soro/química , Soro/efeitos da radiação
9.
Curr Radiopharm ; 6(3): 137-45, 2013 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24106999

RESUMO

A new molecular metallic fragment for labeling biologically active molecules with 99mTc and 188Re is described. This system is composed of a combination of tridentate π-donor and monodentate π-acceptor ligands bound to a [M Ξ N]2+ group (M = (99m)Tc, 188Re) in a pseudo square-pyramidal geometry. A simple structural model of the new metallic fragment was obtained by reacting the ligand 2, 2'-iminodiethanethiol [H2NS2 = NH(CH2CH2SH)2] and monodentate tertiary phosphines with the [M Ξ N]2+ group (M = (99m)Tc, (188)Re). In the resulting complexes (dubbed3+1complexes), the tridentate ligand binds the [M Ξ N]2+ core through the two deprotonated, negatively charged, thiol sulfur atoms and the neutral, protonated, amine nitrogen atom. The residual fourth position of the five-coordinated arrangement is occupied by a phosphine ligand. The chemical identity of these model (99m)Tc and (188)Re compounds was established by comparison with the chromatographic properties of the corresponding complexes obtained at the macroscopic level with the long-lived (99)Tc and natural Re isotopes. The investigation was further extended to comprise a series of ligands formed by simple combinations of two basic amino acids or pseudo-amino acids to yield potential tridentate chelating systems having [S, N, S] and [N, N, S] as sets of π-donor atoms. Labeling yields and in vitro stability were investigated using different ancillary ligands. Results showed that SNS-type ligands afforded the highest labeling yields and the most robust 3+1 nitrido complexes with both (99m)Tc and (188)Re. Thus, the new chelating system can be conveniently employed for labeling peptides and other biomolecules with the [M Ξ N]2+ group.


Assuntos
Quelantes/química , Radioisótopos/química , Compostos Radiofarmacêuticos/química , Rênio/química , Tecnécio/química , Cromatografia Líquida de Alta Pressão , Cromatografia em Camada Fina , Glutationa/química , Humanos , Ligantes , Nitrogênio/química , Compostos de Organotecnécio/química , Peptídeos/química , Soro/efeitos da radiação , Enxofre/química , Fatores de Tempo
10.
Radiat Res ; 179(2): 221-31, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23289387

RESUMO

The biological activities of molecules secreted into the serum of mice chronically irradiated with γ rays at low or medium dose rate (L/MDR) have not been well studied. In this work, the bioactive molecules found in the serum of chronically irradiated mice (dose rate: 0.0181 Gy/h) were characterized by a cell-based assay (CBA) using microarrays. This technique can predict changes in cytokine levels in serum by measuring gene expression profiles and analyzing molecular signaling pathways. Gene expression in cultured mouse embryo fibroblasts (MEFs) 1 day after addition of serum from nonirradiated or irradiated mice had different profiles. A high level of expression of lipocalin2 (Lcn2) was induced in MEFs upon addition of serum from MDR irradiated mice, and Lcn2 was used as a marker for identifying secreted molecules in serum. Based on microarray analysis of molecular pathways, we predicted that the enhanced gene expression of Lcn2 in MEFs might be caused by interleukin-1 (IL-1) in the serum of the irradiated mice, and that an IL-1α antibody could completely neutralize the enhanced gene expression of Lcn2 in MEFs. The increase in IL-1α levels in the serum from the irradiated mice was confirmed by ELISA experiments. However, an increase in IL-1ß could not be detected. These results indicated that IL-1α was released into the serum of mice chronically exposed to a high dose of γ-ray radiation at MDR. We therefore believe that the CBA method using microarrays will be applicable for the screening of bioactive molecules in serum, which will be useful for detecting various diseases and metabolic changes.


Assuntos
Efeito Espectador/efeitos da radiação , Raios gama/efeitos adversos , Proteínas de Fase Aguda/genética , Proteínas de Fase Aguda/imunologia , Animais , Anticorpos Neutralizantes/imunologia , Especificidade de Anticorpos , Efeito Espectador/genética , Linhagem Celular , Citocinas/imunologia , Relação Dose-Resposta à Radiação , Feminino , Lipocalina-2 , Lipocalinas/genética , Lipocalinas/imunologia , Fígado/metabolismo , Fígado/efeitos da radiação , Camundongos , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas Oncogênicas/genética , Proteínas Oncogênicas/imunologia , Soro/metabolismo , Soro/efeitos da radiação , Fatores de Tempo , Transcriptoma/efeitos da radiação
11.
Electromagn Biol Med ; 32(1): 20-9, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23301880

RESUMO

The increasing use of mobile telephones raises the question of possible adverse effects of the electromagnetic fields (EMF) that these phones produce. In this study, we examined the oxidative stress in the brain tissue and serum of rats that resulted from exposure to a 900-MHz EMF at a whole body average specific absorption rate (SAR) of 1.08 W/kg for 1 h/day for 3 weeks. We also examined the antioxidant effect of garlic powder (500 mg/kg/day) given orally to EMF-exposed rats. We found that malondialdehyde (MDA) (p < 0.001) and advanced oxidation protein product (AOPP) (p < 0.05) increased in rat brain tissue exposed to the EMF and that garlic reduced these effects (p < 0.05). There was no significant difference in the nitric oxide (NO) levels in the brain. Paraoxonase (PON) was not detected in the brain. There was a significant increase in the levels of NO (p < 0.001) detected in the serum after EMF exposure, and garlic intake did not affect this increase in NO. Our results suggest that there is a significant increase in brain lipid and protein oxidation after electromagnetic radiation (EMR) exposure and that garlic has a protective effect against this oxidative stress.


Assuntos
Encéfalo/metabolismo , Encéfalo/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Ondas de Rádio/efeitos adversos , Soro/metabolismo , Soro/efeitos da radiação , Produtos da Oxidação Avançada de Proteínas/sangue , Produtos da Oxidação Avançada de Proteínas/metabolismo , Animais , Antioxidantes/farmacologia , Arildialquilfosfatase/sangue , Arildialquilfosfatase/metabolismo , Encéfalo/efeitos dos fármacos , Alho/química , Peroxidação de Lipídeos/efeitos dos fármacos , Peroxidação de Lipídeos/efeitos da radiação , Masculino , Malondialdeído/sangue , Malondialdeído/metabolismo , Óxido Nítrico/sangue , Óxido Nítrico/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Extratos Vegetais/farmacologia , Ratos , Ratos Wistar , Soro/efeitos dos fármacos
12.
Radiats Biol Radioecol ; 52(6): 565-71, 2012.
Artigo em Russo | MEDLINE | ID: mdl-23516887

RESUMO

Using quantitative real-time PCR, the copy number of mitochondrial and nuclear DNA fragments in mouse blood serum was estimated at different time points following X-ray irradiation at various doses (from 0.5 to 10 Gy). The changes in the correlation between mtDNA and nuclear DNA (mtDNA/nucDNA) in blood serum reflect the degree of radiation injury depending on the dose of irradiation. Exposure to radiation at 10 Gy and massive cell death caused by this lethal dose result in a sharp decrease by an order of magnitude of the mtDNA/nucDNA ratio in the mouse serum; the value of this parameter did not recover within the next 3 days. The opposite effect was revealed when mice were exposed to irradiation at the dose of I Gy, which is not followed by massive cell death, but leads to a higher level of the mtDNA damage as compared with the nuclear DNA protected by histones. Defective mtDNA molecules enter the bloodstream, which results in an increase of the mtDNA/nucDNA ratio in serum. Under irradiation of mice at the intermediate dose of 3 Gy the two processes described above are exhibited at once. During the first hours after irradiation an apoptotic death of radiosensitive cells and release of a large number of nuclear DNA fragments in the serum are initiated, which reduces the mtDNA/nucDNA ratio. However, at later times after irradiation, starting from 5 days, an increase of the mtDNA/nucDNA ratio is observed in the serum, presumably as a result of reparation and elimination of defective mtDNA. Thus, the mtDNA/nucDNA ratio in the serum of irradiated mice reflects the degree of the radiation damage to cells and may be considered as a biological marker of radiation injury in the future.


Assuntos
DNA Mitocondrial , DNA , Soro , Animais , DNA/sangue , DNA/efeitos da radiação , DNA Mitocondrial/sangue , DNA Mitocondrial/efeitos da radiação , Relação Dose-Resposta à Radiação , Camundongos , Soro/citologia , Soro/efeitos da radiação , Raios X
13.
Biologicals ; 39(6): 370-7, 2011 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21871817

RESUMO

Animal-derived materials such as animal sera represent a low, but finite, risk for introduction of an adventitious agent (virus or mollicute) into a biological bulk harvest during upstream manufacturing processes involving mammalian cell substrates. Viral and mollicute (Mycoplasma sp. and Acholeplasma sp.) contamination events have been relatively rare, but many of those that have been reported have been attributed to use of infected animal sera in growth media during cell expansion. The risk of introduction of viruses and mollicutes may be mitigated by elimination of the use of animal sera and implementation instead of chemically defined or serum- and animal-derived material-free cell culture media. When use of animal sera is unavoidable, however, mitigation of the risk of introducing an adventitious contaminant may involve treatment of the sera to inactivate potential contaminants. Gamma irradiation is one of the most widely employed methods for viral and mollicute inactivation in animal sera. In this article, we review the inactivation results reported for viral and mollicute inactivation in frozen serum. Studies performed to assess the impact of gamma irradiation on serum quality and performance are also discussed. The available data indicate that inactivation of mollicutes in serum is essentially complete at the gamma radiation doses normally employed (25-40 kGy), while the efficacy and kinetics for viral inactivation in serum by gamma irradiation appear to be dependent in part upon the size of the target virus.


Assuntos
Acholeplasma/efeitos da radiação , Raios gama , Mycoplasma/efeitos da radiação , Soro/efeitos da radiação , Vírus/efeitos da radiação , Animais , Meios de Cultura/química , Meios de Cultura/efeitos da radiação , Relação Dose-Resposta à Radiação , Contaminação de Medicamentos/prevenção & controle , Soro/microbiologia , Soro/virologia , Inativação de Vírus/efeitos da radiação
14.
Radiats Biol Radioecol ; 51(1): 76-80, 2011.
Artigo em Russo | MEDLINE | ID: mdl-21520619

RESUMO

The purpose of this work was the analysis of the effects of bystander factors from blood sera of people affected by the Chernobyl accident on human keratinocyte cell culture (HPV-G cells). A new method was developed for evaluation of the bystander factor presence in vivo in blood of the people irradiated by the Chernobyl accident. Affected population groups included liquidators of the Chernobyl accident and people living and working in areas of the Gomel region contaminated by radionuclides. The analysis has shown that bystander factors persist in Chernobyl liquidator blood samples for more than 20 years since irradiation. The data suggest that blood sera contain bystander factors, which are able to induce micronuclei and decrease the metabolic activity of HPV-G cells.


Assuntos
Fatores Biológicos/farmacologia , Efeito Espectador/genética , Acidente Nuclear de Chernobyl , Soro/efeitos da radiação , Fatores Biológicos/sangue , Estudos de Casos e Controles , Técnicas de Cultura de Células , Linhagem Celular , Humanos , Queratinócitos/efeitos dos fármacos , Queratinócitos/metabolismo , Queratinócitos/ultraestrutura , Melaninas/farmacologia , Melatonina/farmacologia , Micronúcleos com Defeito Cromossômico/efeitos da radiação , Testes para Micronúcleos , Protetores contra Radiação/farmacologia , Soro/química , Ucrânia
16.
Tsitologiia ; 52(9): 785-92, 2010.
Artigo em Russo | MEDLINE | ID: mdl-21105368

RESUMO

Simultaneous low-intensity visible (VIS) and near infrared (nIR) irradiation from laser and non-laser sources was used for treatment of complications developing in cancer patients after surgical tumor resection, chemo- and radiation therapy. However, the question remains about the impact of this physiotherapeutic method on proliferative activity of the patients' tumor cells and cells involved in wound healing, fibroblasts (FB) and keratinocytes (KC). In this paper, we studied the effect blood serum obtained from the patients with breast cancer after the course of irradiation with visible and NI light (480--3400 nm, 95 % polarization, 40 mW/cm2, 12 J/cm2) in postoperative period on the proliferative activity of primary cultures of human FB and KC, and of several human tumor cell lines (BT-474, HBL-100, Hs578T and A431). Seven-day course of phototherapy increase proliferation of FB (as compared to the initial level) and KC (as compared to postoperative level) by 22 and 28 %, respectively. The tumor cells BT-474, Hs578T and A431 showed statistically significant decrease in proliferative activity compared with the preoperative (initial) level by 31.5, 8.97 and 6.47%, respectively, whereas the cells BT-474, HBL-100, Hs578T and A431 also reduced their proliferative activity by 32,16, 8.65 and 6.26%, respectively, as compared with postperative level. The results obtained demonstrate the safety of the phototherapy with the visible and NI light for BC patients in the postoperative period.


Assuntos
Neoplasias da Mama/patologia , Neoplasias da Mama/terapia , Proliferação de Células , Fototerapia/métodos , Soro/efeitos da radiação , Neoplasias da Mama/sangue , Células Cultivadas , Feminino , Fibroblastos/citologia , Fibroblastos/patologia , Humanos , Raios Infravermelhos , Queratinócitos/citologia , Queratinócitos/patologia , Luz , Pessoa de Meia-Idade , Células Tumorais Cultivadas
17.
PLoS One ; 5(4): e10343, 2010 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-20436679

RESUMO

The culture and demonstration of putative nanobacteria (NB) and calcifying nanoparticles (CNP) from human and animal tissues has relied primarily on the use of a culture supplement consisting of FBS that had been gamma-irradiated at a dose of 30 kGy (gamma-FBS). The use of gamma-FBS is based on the assumption that this sterilized fluid has been rid entirely of any residual NB/CNP, while it continues to promote the slow growth in culture of NB/CNP from human/animal tissues. We show here that gamma-irradiation (5-50 kGy) produces extensive dose-dependent serum protein breakdown as demonstrated through UV and visible light spectrophotometry, fluorometry, Fourier-transformed infrared spectroscopy, and gel electrophoresis. Yet, both gamma-FBS and gamma-irradiated human serum (gamma-HS) produce NB/CNP in cell culture conditions that are morphologically and chemically indistinguishable from their normal serum counterparts. Contrary to earlier claims, gamma-FBS does not enhance the formation of NB/CNP from several human body fluids (saliva, urine, ascites, and synovial fluid) tested. In the presence of additional precipitating ions, both gamma-irradiated serum (FBS and HS) and gamma-irradiated proteins (albumin and fetuin-A) retain the inherent dual NB inhibitory and seeding capabilities seen also with their untreated counterparts. By gel electrophoresis, the particles formed from both gamma-FBS and gamma-HS are seen to have assimilated into their scaffold the same smeared protein profiles found in the gamma-irradiated sera. However, their protein compositions as identified by proteomics are virtually identical to those seen with particles formed from untreated serum. Moreover, particles derived from human fluids and cultured in the presence of gamma-FBS contain proteins derived from both gamma-FBS and the human fluid under investigation-a confusing and unprecedented scenario indicating that these particles harbor proteins from both the host tissue and the FBS used as feeder. Thus, the NB/CNP described in the literature clearly bear hybrid protein compositions belonging to different species. We conclude that there is no basis to justify the use of gamma-FBS as a feeder for the growth and demonstration of NB/CNP or any NB-like particles in culture. Moreover, our results call into question the validity of the entire body of literature accumulated to date on NB and CNP.


Assuntos
Técnicas Bacteriológicas/métodos , Proteínas Sanguíneas/efeitos da radiação , Raios gama/efeitos adversos , Soro/efeitos da radiação , Bactérias/citologia , Cálcio , Humanos , Nanopartículas , Soro/microbiologia , Esterilização/métodos
19.
Radiats Biol Radioecol ; 50(1): 28-36, 2010.
Artigo em Russo | MEDLINE | ID: mdl-20297678

RESUMO

This study evaluated possible adverse effects of injection of blood serum from rats exposed to microwaves at a power density of 500 microW/cm2 on pregnancy and foetal and offspring development in intact female rats. The study was performed with 59 pregnant Wistar rats. In utero mortality, embryo and foetal body weights and placenta weight were used for the evaluation of embryo and foetal development. Generally accepted integral and specific parameters were used for the evaluation of postnatal development of offspring during the first 30 days of life. It was shown that intra peritoneal injection of blood serum from IMF exposed rats (chronic 30-day RF exposure at 500 microW/cm2) to intact rats on the 10th day of pregnancy resulted in adverse effects on foetal and offspring development. Total mortality (in utero + postnatal) as well as delay in offspring development was higher in this group.


Assuntos
Anormalidades Induzidas por Radiação/sangue , Campos Eletromagnéticos/efeitos adversos , Feto/efeitos da radiação , Complicações na Gravidez/sangue , Reprodução/efeitos da radiação , Soro/efeitos da radiação , Anormalidades Induzidas por Radiação/imunologia , Animais , Animais Recém-Nascidos , Feminino , Feto/imunologia , Relações Materno-Fetais , Gravidez , Complicações na Gravidez/imunologia , Ratos , Ratos Wistar , Soro/imunologia
20.
Biologicals ; 38(1): 135-43, 2010 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-19716317

RESUMO

Effects of 18 commercial lots of fetal calf serum (FCS) after gamma-irradiation and their non-irradiated counterparts were comparatively analyzed on CHO-K1 and MDBK MDL1 cells for genotoxicity [sister chromatid exchange (SCE), micronuclei (MNi), and single cell gel electrophoresis (SCGE)], cytotoxicity [cell-cycle progression (CCP), proliferative replication index (PRI), mitotic index (MI), growth promotion (GP), and plating efficiency (PE)], and microbiological properties (mycoplasma and bovine viral diarrhea virus contamination). SCE and SCGE were the most informative end-points for genotoxicity since significant differences were found in 44.4% (P<0.05-0.001, Student's t-test) and 61.1% (P<0.05-0.001, chi(2) test) samples, respectively. MI was the cytotoxicity assay revealing the greatest variation, showing differences in 66.7% (P<0.05-0.001, chi(2) test) samples. Thus, these three end-points for screening bioproducts such as FCS were found most suitable for detecting potential geno-cytotoxicants in biological samples; their simultaneous use could be strongly recommended.


Assuntos
Sangue Fetal/fisiologia , Esterilização , Testes de Toxicidade/métodos , Animais , Patógenos Transmitidos pelo Sangue/efeitos da radiação , Células CHO , Bovinos , Técnicas de Cultura de Células/métodos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Comércio , Cricetinae , Cricetulus , Meios de Cultura/farmacologia , Vírus da Diarreia Viral Bovina/efeitos da radiação , Cães , Sangue Fetal/efeitos da radiação , Testes de Mutagenicidade , Soro/fisiologia , Soro/efeitos da radiação , Esterilização/métodos
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