ABSTRACT
@#Objective To develop and verify a whole column imaging detection-capillary isoelectric focusing(WCID-CIEF)method for the determination of isoelectric point(pI)of pertactin(PRN).Methods The WCID-CIEF method for the determination of PRN antigen was developed by optimizing the parameters such as the focusing time and final concentration of samples in the WCID-CIEF process,and verified for the specificity,accuracy,repeatability,intermediate precision,durability and inter-batch consistency.Results The optimal focusing time of WCID-CIEF for the determination of PRN antigen pI was 1 min at 1 500 V and 3 min at 3 000 V. The optimal final concentration of PRN antigen was 300 μg/mL. The PRN antigen pI was about 6. 035,and the blank matrix showed no interference peak in the position of each peak of antigen. The method had good specificity,accuracy,repeatability,intermediate precision,durability and consistency among batches.Conclusion The developed WCID-CIEF method is suitable for the pI detection and charge heterogeneity analysis of PRN antigen,which can provide basis for the characterization of PRN antigen and reference for the quality control in the process of development and production of related vaccines.
ABSTRACT
Existing neuroregulatory techniques can achieve precise stimulation of the whole brain or cortex, but high-focus deep brain stimulation has been a technical bottleneck in this field. In this paper, based on the theory of negative permeability emerged in recent years, a simulation model of magnetic replicator is established to study the distribution of the induced electric field in the deep brain and explore the possibility of deep focusing, which is compared with the traditional magnetic stimulation method. Simulation results show that a single magnetic replicator realized remote magnetic source. Under the condition of the same position and compared with the traditional method of stimulating, the former generated smaller induced electric field which sharply reduced with distance. By superposition of the magnetic field replicator, the induced electric field intensity could be increased and the focus could be improved, reducing the number of peripheral wires while guaranteeing good focus. The magnetic replicator model established in this paper provides a new idea for precise deep brain stimulation, which can be combined with neuroregulatory techniques in the future to lay a foundation for clinical application.
Subject(s)
Brain , Cerebral Cortex , Computer Simulation , Electricity , Magnetic FieldsABSTRACT
@#Objective To develop and verify a whole-column image capillary isoelectric focusing(iCIEF) electrophoresis method to analyze the charge heterogeneity of recombinant human growth hormone Fc fusion protein(Fc-rhGH).Methods The iCIEF analysis method of Fc-rhGH was developed by optimizing the target protein concentration,cosolvent(urea)concentration and focusing time.The target protein was simultaneously analyzed by this method and traditional flat plate isoelectric focusing(IEF) electrophoresis,and the results were compared;The specificity,accuracy,precision,limit of quantitation(LOQ) and durability of the developed method were verified.Results The optimized method was using the mixed solution of 8 mol/L urea,0.35% methyl cellulose(MC),4% amphoteric electrolyte and 0.5% isoelectric point marker as the sample buffer,and the focusing condition was 1 500 V 1 min,3 000 V 5.5 min.IEF was not suitable for analyzing the charge heterogeneity of Fc-rhGH solution.Using the optimized iCIEF for analysis,the target protein was significantly different from the unrelated protein,and the baseline of blank reagent was stable;The recovery rate of accuracy verification was within 90%～110%,and the linear range was 0.25～0.75 mg/mL(50%～150% of the target loading volume);The RSD of each isomer pI in the repeatability verification was less than 0.3%,and the RSD of peak area percentage was less than 5%;The LOQ was 0.04 mg/ml.The sample storage time durability,amphoteric electrolyte pharmalyte 3-10 durability and MC durability of this method were good.Using this method to analyze the charge heterogeneity of Fc-rhGH physicochemical reference substance,eight charge heterogeneities of the reference substance were effectively separated,and the pI ranged from 5.9 to 6.4.Conclusion The developed iCIEF method had good specificity,accuracy,precision and durability,and was more suitable for efficient analysis of charge heterogeneity of Fc-rhGH than traditional flat plate IEF,which was of great significance for the quality control of Fc-rhGH and other Fc fusion proteins.
ABSTRACT
@#Objective To establish and verify a capillary isoelectric focusing-whole column imaging detection(CIEFWCID) method for the analysis of isoelectric point(pI) of recombinant human brain natriuretic peptide.Methods The ampholyte,space-occupying agent,protein concentration,focusing time were optimized by CIEF-WCID method,and the best condition for the detection of recombinant human brain natriuretic peptide was obtained.The repeatability,precision and durability of the developed method were verified,and three batches of recombinant human brain natriuretic peptide produced continuously were analyzed for pI.Results HR AESlyte 8-10.5 was selected as ampholyte,while 25 mmol/L sodium hydroxide as the space-occupying agent;The final concentration of the sample was 87.5 μg/mL and the focusing time was 8min.The relative standard deviation RSD of pI detection was 0.1% after six consecutive injections of the same sample;The RSD of pI detection of six samples was 0.1%;The pI RSD of the main peak was 0.1% at different final concentrations of the sample,and the pI RSD of the sample was 0.1% at different storage time,while the pI markers could not be changed arbitrarily.The pI was detected in three consecutive batches of recombinant human brain natriuretic peptide samples.Conclusion The developed CIEF-WCID method for pI analysis of recombinant human brain natriuretic peptide had good repeatability and precision and might be used for follow-up quality control of recombinant human brain natriuretic peptide.
ABSTRACT
@#Ultrasound cycloplasty(UCP)procedure using high intensity focused ultrasound(HIFU)is a new non-invasive glaucoma treatment based on miniaturized transducers. In recent years, the safety and efficacy of UCP on various types of glaucoma have been well verified, and it has shown broad application prospects in the treatment of glaucoma. This article collects and reviews the relevant literature on glaucoma treatment by UCP, and summarizes the information on the development of equipment, composition and operation, treatment mechanisms, indications, efficacy and safety, with the purpose to provide relevant basis for clinical applications and future research.
ABSTRACT
Isoelectric focusing and chromatofocusing are techniques to separate amphoteric compounds based on their isoelectric points. The commonest used c ompounds to generate a pH gradient in these techniques are ampholytes. The prohibitively expensive cost of carrier ampholytes limits the application of these techniques in less advantaged parts of the world. A simple and inexpensive method is de scribed in this paper to prepare a broad pH range (pH 3 to 11) ampholytes for use in isoelectric focusing or chromatofocusingin any routine laboratory. The method involves crosslinking polyethylene polyamine to create complex cross-linked polyaminesin the first step. Whereas, the second step involves reacting the cross-linked polyamines with an unsaturated carboxylic acid to produce ampholytes.
ABSTRACT
Resumo Observar e analisar as causas, os efeitos e as múltiplas consequências econômicas, sociais e sanitárias da pandemia da COVID-19 têm sido primordiais não apenas para a compreensão desse fenômeno mundial, mas também para a elaboração de alternativas e soluções que minimizem os impactos na vida das populações ao redor do mundo. Este artigo tem como principal objetivo entender a ascensão da renda básica emergencial como alternativa viável de política pública no cenário brasileiro, por meio do Auxílio Emergencial. Baseado na literatura de agenda-setting, em específico no conceito de eventos focalizadores (focusing events), recuperamos um breve histórico das políticas de transferência de renda e de combate à pobreza e os principais atores envolvidos, para entender mudanças, adaptações e soluções propostas para que a alternativa de renda básica emergencial pudesse ser considerada e aceita neste momento específico de crise.
Resumen Observar y analizar las causas, efectos y las múltiples consecuencias económicas, sociales y de salud de la pandemia de COVID-19 han sido primordiales no solo para comprender este fenómeno mundial, sino también para el desarrollo de alternativas y soluciones que minimicen los impactos en la vida de poblaciones de todo el mundo. El principal objetivo de este artículo es entender el reconocimiento de la renta básica de emergencia como una alternativa viable de política pública en el escenario brasileño, que ha adoptado un "beneficio de emergencia". Con base en la literatura de agenda-setting, específicamente en el concepto de eventos focales, recuperamos una reseña de las políticas de transferencia de renta, de reducción de la pobreza y los principales actores involucrados, para comprender los cambios, adaptaciones y soluciones propuestas para que la alternativa de renta básica de emergencia pudiera considerarse y aceptarse en este momento específico de crisis.
Abstract Observing and analyzing the causes, effects, and multiple economic, social, and health consequences of the COVID-19 pandemic have been essential to understand this worldwide phenomenon and find solutions that minimize the impacts on peoples' lives. This article aims to understand the rise of the emergency basic income as a viable policy to tackle the pandemic in Brazil. Based on the agenda-setting literature, specifically on the concept of focusing events, we present a brief history of cash transfer and poverty reduction policies, as well as the actors involved. The study discusses the changes, adaptations, and solutions proposed during the formulation of the emergency basic income as a policy to be considered and effectively adopted in this specific moment of crisis.
Subject(s)
Public Policy , Brazil , Pandemics , COVID-19 , Government , IncomeABSTRACT
Abstract Objectives: To characterize cases of suspected congenital disorders of glycosylation (CDG) investigated in a laboratory in southern Brazil using the transferrin isoelectric focusing TfIEF test from 2008 to 2017. Method: Observational, cross-sectional, retrospective study. The laboratory records of 1,546 individuals (median age = 36 months, 25-75 IQR = 10-108; males = 810) submitted to the TfIEF test during the period were reviewed. Results: Fifty-one individuals (3%) had an altered TfIEF pattern (5 ± 2.8 cases/year; median age = 24 months, 25-75 IQR = 11-57 months; males = 27, 53%). For 14 of them, data on diagnosis conclusion were available (classic galactosemia = 4; hereditary fructose intolerance = 4; peroxisomal diseases = 2; PMM2-CDG = 2; MPDU1-CDG = 1; SLC35A2-CDG = 1).Comparing the cases with the normal and altered TfIEF patterns, there was a higher prevalence of altered cases in the age group from 11 months to 3 years. There was an increase in the likelihood of change in TfIEF, especially in the presence of inverted nipples or liver disease. Conclusions: The data suggest that the investigation of a case with suspected CDG is a complex problem, being aggravated by the existence of other IEMs (inborn errors of metabolism) associated with altered TfIEF pattern and lack of access to confirmatory tests. The presence of inverted nipples and liver disease, especially in individuals aged 11 months to 3 years, should suggest the need for TfIEF investigation.
Resumo Objetivos: Caracterizar os casos com suspeita de CDG investigados em laboratório do sul do Brasil pelo exame de IEFTF de 2008 a 2017. Metodologia: Estudo observacional, transversal, retrospectivo. Foram revisadas as fichas laboratoriais de 1.546 indivíduos (mediana de idade = 36 meses, IQ 25-75 = 10-108; sexo masculino = 810) que fizeram o exame de IEFTF no período. Resultados: Cinquenta e um indivíduos (3%) apresentaram padrão alterado na IEFTF (5 ± 2,8 casos/ano; mediana de idade = 24 meses, IQ 25-75 = 11-57 meses; sexo masculino = 27, 53%). Para 14 deles, estavam disponíveis dados sobre a conclusão do diagnóstico (galactosemia clássica = 4; intolerância hereditária à frutose = 4; doenças peroxissomais = 2; PMM2-CDG = 2; MPDU1-CDG = 1; SLC35A2-CDG = 1). Comparando os casos com padrão normal e alterado na IEFTF, houve maior prevalência de casos alterados na faixa etária de 11 meses a 3 anos. Verificou-se um aumento na probabilidade de alteração na IEFTF principalmente na presença de mamilos invertidos ou de hepatopatia. Conclusões: Os nossos dados sugerem que a investigação de um caso com suspeita de CDG é complexa, é agravada pela existência de outros EIM associados a padrão alterado na IEFTF e pela falta de acesso a exames confirmatórios. A presença principalmente de mamilos invertidos e de hepatopatia em indivíduos na faixa etária de 11 meses a 3 anos deve sugerir a necessidade de investigação por IEFTF.
Subject(s)
Humans , Infant , Transferrin/analysis , Congenital Disorders of Glycosylation/diagnosis , Congenital Disorders of Glycosylation/epidemiology , Isoelectric Focusing , Brazil , Cross-Sectional Studies , Retrospective StudiesABSTRACT
OBJECTIVE: To analyze the glycosylated chains of recombinant interleukin-15 fusion protein using capillary isoelectric focusing-whole column imaging detection (WCID-cIEF) spectrograms. METHODS: Using established corresponding mathematical models and the least square method, the WCID-cIEF spectrograms of whole protein, de-salicylic-acid protein and de-N-glycosylation-chain protein were analyzed. Among the mathematical models, the interval-1-peak model was selected. And according to the model, the relationship between isoform peak-areas and isoelectric points was listed. RESULTS: The rationality of the interval-1-peak model was confirmed and a series of basic data was obtained according to the model as follows:the apparent m value of the protein was 25.53 reference(R), the apparent n value of the protein was 28.83R, the apparent m value of sialic acid was 0.86 (0.855) R, the apparent n value was 0.12 (0.119) R, the apparent n value of N-acetylglucosamine (undifferentiated from N-acetylgalactosamine) was 0.06(0.061) R, and the apparent m value of formed carboxyl after N-chain removal was 0.19 (0.186) R. Some information of protein sugar composition was also obtained: the sialylation degree was about 1.83 mol•mol-1, the percentage of prototype protein was about 8.3%, the percentage of single N-glycosated modification protein was about 19.8%, the percentage of double N-glycosated modification protein was about 28.4%, the percentage of triple N-glycosated modification protein was about 23.7%, and the percentage of O-glycosated modification (with sialic acid) protein was about 19.8%. The main sugar types should be G0 (F), G1 (F), G2 (F), G1A1 (F), and G2A1 (F). CONCLUSION: The structure of sugar chain is complex, but it also has some repeatability and regularity. We hope that through this study, the glycoprotein sugar chain can be quickly outlined, the understanding of glycoprotein and the study of protein interaction can be improved.
ABSTRACT
In this study, we sought to describe a novel imaging apparatus that is lightweight, inexpensive, and highly eff ective for use in colorectal diagnostic and treatment settings. Typical probes for use in colorectal ultrasonic imaging applications are developed for surgeons to diagnose and stage rectal tumors and image the rectum and anus. Here we outline a new technique and use it for colorectal imaging in an animal. This technique involves use of an ultrasound array module positioned along the axis of rotation such that improved rotation is possible. This module is in the shape of a linear rod with a rotary linear component that allows for emission of focused ultrasonic echo signals from a linear section of the probe. The usability of the transducer and rectal image quality are satisfactory in a porcine model with the technique proposed here, axial/lateral resolution as 0.96/2.24 mm with 6 dB applied through the contour map using the point spread function. When compared to currently available methods, this technique provides superior diagnostic 3D volumetric image quality with reduced acquisition time. Given this, the ultrasound device proposed here may prove a viable and preferable method to those currently available for urology and colorectal imaging applications.
Subject(s)
Animals , Anal Canal , Methods , Rectal Neoplasms , Rectum , Surgeons , Transducers , Ultrasonics , Ultrasonography , UrologyABSTRACT
PURPOSE: Focusing manner is very powerful mind-body self-help and self-awareness competence. So focusing manner may affect nurses' emotional labor and nursing performance, but few are known about it to date. The purpose of this study was to identify the degree of focusing manner, emotional labor, nursing performance of clinical nurses, and the factors that affect nursing performance. METHODS: In order to collect data, structured questionnaires were administered to 212 nurses who worked at 2 university hospitals located in D city. Data were analyzed by t-test, ANOVA, correlation and multiple regression using SPSS/WIN 20.0. RESULTS: Nursing performance had positive relationship with focusing manner (r=.45, p<.001), and negative relationship with emotional labor (r=−.25, p<.001). And the most prediction factor influencing nursing performance was focusing manner (β=.41, p<.001) and the other factors were age (β=.30, p=.002), and emotional labor (β=.14, p=.012). The total variance was 37.1% by those predictors (F=25.87, p<.001). CONCLUSION: Based on the results of this study, it is necessary to develop and educational program on focusing manner-oriented psychotherapy in order to improve the nursing performance. Also, the management system for controlling emotional labor needs to be set up for nurses in clinical settings.
Subject(s)
Hospitals, University , Mental Competency , Nursing , PsychotherapyABSTRACT
The present situation and research progress of the single-particle focusing device and related technologies were introduced in foreign countries and China. The principles and advantages of different single-particle focusing devices were analyzed from the aspects of liquid and gas single-particle focusing, and it's pointed out that rapid detection of the particles such as bacteria could be realized by liquid and gas single-particle focusing measures. The two measures both had brilliant prospects providing the structure and design were optimized to ensure high detection precision and efficiency, which could be promoted for bio-agents detection and fulminating infectious diseases prevention and treatment.
ABSTRACT
O presente estudo objetivou investigar as relações entre religiosidade e autoconsciência situacional e disposicional, fundamentando-se na hipótese de uma possível coevolução conjunta na ontogênese em estudantes universitários de distintas orientações religiosas. A amostra foi composta por 958 universitários (52,1% de sexo feminino), os quais responderam ao protocolo de pesquisa contendo Questionário Sociodemográfico, Escala de Autoconsciência Situacional, Escala de Autoconsciência Disposicional e Escala de Religiosidade Global, e os dados foram analisados através de Análise Fatorial, Anova One-Way seguida do teste das diferenças honestamente significativas (DHS) de Tukey e coeficiente de correlação de Pearson. Os achados levantaram evidências de uma sistemática interrelação entre fatores de autoconsciência situacional e disposicional e dimensões de religiosidade, corroborando a hipótese de um papel protetivo da religiosidade ao desenvolvimento dos processos autofocalizadores humanos
The present study aimed to investigate the relationship between religiosity and situational and dispositional self-focus, based on the hypothesis of a possible joint co-evolution in ontogeny in university students of different religious orientations. The sample consisted of 958 college students (52.1% females), which responded to the research protocol containing Sociodemographic Questionnaire, Situational Self-focus Scale, Dispositional Self-focus Scale and Global Religiosity Scale. Data were analyzed using factor analysis one-way ANOVA, followed by the Tukey's Honestly Significantly Different (DHS) test and Pearson correlation coefficient. The findings raised evidence of a systematic interrelation between factors of situational and dispositional self-focus and dimensions of religiosity, supporting the hypothesis of a protective role of religiosity in the development of human self-focused attention processes
El presente estudio está enfocado a la investigación de las relaciones entre religiosidad y autoconsciencia situacional y de disposición, basado en la hipótesis de una posible coevolución conjunta en la ontogénesis en estudiantes universitarios de diversas orientaciones religiosas. La muestra está compuesta de 958 universitarios (52,1% de sexo femenino), que respondieron al protocolo de investigación que contiene el Cuestionario Sociodemográfico, Escala de Autoconsciencia Situacional, Escala de Autoconsciencia de Disposición y Escala de Religiosidad Global, se analizaron los datos por medio de Análisis Factorial, ANOVA One-Way seguida del test de diferencias honestamente significativas (DHS) de Tukey y el coeficiente de correlación de Pearson. Los hallazgos mostraron evidencias de una interrelación sistemática entre factores de autoconsciencia situacional y de disposición y dimensiones de religiosidad, corroborando la hipótesis de un papel protector de la religiosidad en el desarrollo de procesos auto enfocadores humanos
Subject(s)
Humans , Cognition , Conscience , Religion and Psychology , SpiritualityABSTRACT
Este texto debruça-se sobre uma nova forma de fazer psicoterapia psicanalítica e psicanálise. A exploração das emoções como meio de mudança. Relato uma alteração na minha prática clínica em que a emoção passou a ser o foco da atenção. Uso um maior equilíbrio entre guiar e seguir o paciente, uma atenção aos marcadores e uso das intervenções para cada um desses marcadores como o desdobrar sistemático de uma evocação, focalização, ou o diálogo da cadeira vazia. São abordadas algumas dificuldades de integrar uma exploração mais eficaz da emoção num modelo psicanalítico praticado há muitos anos. Pequenas vinhetas clínicas ilustram a forma de explorar as emoções.
This text is about a new way of doing psychoanalytic psychotherapy and psychoanalysis. The exploration of emotions as a means of change. I report a change in my clinical practice in which emotion has become the focus of attention. I use a greater balance between guiding and following the patient, an attention to the markers and use of interventions for each of these markers as the systematic evocation unfolding, focusing, or the empty-chair dialogue. I describe some difficulties of integrating a more effective exploration of the emotion in a psychoanalytic model practiced for many years. Small clinical vignettes illustrate how to explore emotions.
ABSTRACT
Objective]Discussion on academic ideas and clinical experience of Professor Wang Xu in treating of throid-associated ophthalmopathy from liver, spleen and kidney.[Methods]From the relationship of etiology and pathogenesis, clinical manifestation, prognosis and therapy with liver, spleen and kidney to expound the academic viewpoints and clinical experience of Professor Wang Xu in treatment of TAO. The course of TAO is divided into early stage, middle stage and advanced stage. Early stage should be treated from the liver, middle stage should be treated from the spleen, and advanced stage should be treated from the kidney.In the end, we examplify that. [Results] Professor Wang Xu believes the liver, spleen and kidney in Chinese Medicine sense have close relationship on TAO. The pathogenesis of this disease is “Liver stagnation transforming into fire and phlegm, phlegm fire disturbing orifice and damaging liver and kidney after a long time”, phlegm and blood stasis are the important pathological factors. Thus, the treatment of TAO focusing on liver, spleen and kidney is proposed. Clearing liver and nourishing liver yin, strengthening the spleen and removing dampness, dissipating phlegm and activating blood, nourishing Yin of liver and kidney is the clinical common therapy, clinically obtaining better therapeutic effect. [Conclusion] The clinical experience of Professor Wang Xu in treatment of TAO is effective, has the value of popularization and application.
ABSTRACT
Objective To investigate the effects of submarine environment on the ability of attention focusing and distribution of submariners through a comparative study. Methods To measure simple reaction time, complex reaction time, correct times of reaction, the time on targets as well as the times out of track with attention focus measuring instrument (EP701C) and attention distribution measuring instrument(EP708A). Results In the attention focusing test, the times out of track of target was significantly more in submariners with a 3-5 years service experience compared with their off-shore counterparts(t=2.03,P<0.05). The time on targets was significantly shorter in submariners with a more than 5 years service experience compared with their off-shore counterparts(t=2.27,P<0.05), and the times out of track of target was significantly more in submariners (t=2.12,P<0.05). In the attention distribution test, the correct times of reaction in submariners with 3-5 years service was significantly less than their off-shore counterparts(Z=7.03,P<0.01), and the same was also true for those with a more than 5 years service experience(Z=7.74,P<0.01). Conclusion Our findings indicate that the attention abilities of submariners are lower than those of off-shore staffs, and 3-5 year service period is the important time for impairment of attention ability among submariners.
ABSTRACT
Objective To compare the single-shot fields irradiated by three focusing modes of γ-knife and explore the approaches for improving the quality of stereotactic radiosurgery.Methods GAFCHROMIC(R) EBT3 mode flushing-free film was used to measure the single-shot fields irradiated by multi-source static focusing modes,multi-source single-axis rotating focusing mode and single-source double-axis rotating focusing mode of γ-knife.Also the uniformity and penumbra of the single-shot fields were compared.Results The 2D dose distribution of the single-shot fields irradiated by three focusing modes of γ-knife was different.In the axis (x,y,z),the rang of penumbra axial length ratios of multisource static focusing modes,multi-source single-axis rotating focusing mode and single-source double-axis rotating focusing mode were 0.13-0.48,0.17-0.33 and 0.28-0.54,in the diagonal direction of the wings plane (NSD,PSD),were 0.31-0.39,0.38-0.43 and 0.54-0.72,respectively;the penumbra axial length ratio of single-source double-axis rotating focusing mode was bigger than in multi-source static focusing modes and multi-source single-axis rotating focusing mode.On the no-wings plane,the area ratios of 80％ dose curve enveloped and 50％ dose curve enveloped(A80％/A50％)were 0.40,0.47 and 0.19,on the wings plane,were 0.61,0.53 and 0.35,respectively.The field uniformity of multi-source static focusing modes and multi-source single-axis rotating focusing mode were superior to single-source doubleaxis rotating focusing mode.Conclusions Considering dose distribution of the single-shot fields,the multi-source static focusing modes devices and the multi-source single-axis rotating focusing mode devices should be preferred,when important tissues and organs are adjacent to the target areas.Compared with single-source double-axis rotating focusing mode,both multi-source static focusing modes and multi-source single-axis rotating focusing mode could make more target areas to be surrounded by high dose region.
ABSTRACT
OBJECTIVE: The aim of this study is to determine a protocol of gingival crevicular fluid protein extraction used for the first dimension of 2-DE gels. It also aims at conducting a review on the current candidates for protein markers of this pathology, all of which may be used to prevent the disease. METHODS: Gingival crevicular fluid was collected from two groups of 60 patients each, with and without external root resorption. Samples were extracted by means of various methods of protein extraction. SDS-PAGE gels were used to assess the quality of the method which was subsequently tested during isoelectric focusing of 2-DE gels taken from samples of patients with and without the disease. RESULTS: Milli-Q ultrapure ice cold water, without precipitation for gingival crevicular fluid protein extraction, proved the method with greatest sharpness to detect protein bands. Additionally, it allowed two-dimensional electrophoresis to be performed. CONCLUSION: The new protein extraction protocol does not interfere in isoeletric focusing of 2-DE gels. Furthermore, it provides the greatest sharpness in detecting protein bands of SDS-PAGE gels. This will allow mapping and searching of new external root resorption markers, particularly due to the difficulty in carrying out molecular tests with the current candidates for protein markers. .
OBJETIVO: o objetivo desse trabalho foi determinar o protocolo de extração proteica do fluido crevicular gengival, que pudesse ser utilizado para a realização da primeira dimensão dos géis 2-DE, bem como fazer uma revisão dos atuais candidatos a marcadores proteicos dessa patologia que podem ser utilizados na prevenção dessa doença. MÉTODOS: foi coletado o fluido crevicular gengival de dois grupos de 60 pacientes, com e sem a reabsorção radicular externa. As amostras foram extraídas por diversos métodos de extração proteica e utilizados géis SDS-PAGE para aferir a qualidade do método, que posteriormente foi testado durante a realização da focalização isoelétrica dos géis 2-DE, de amostras de pacientes com e sem a patologia. RESULTADOS: a utilização de água Milli-Q gelada ultrapura, sem nenhuma precipitação para a extração proteica do fluido crevicular gengival, foi o método com maior nitidez das bandas proteicas, além de permitir a realização da eletroforese bidimensional. CONCLUSÕES: o novo protocolo de extração proteica não interfere na focalização durante a realização dos géis 2-DE, além de maior nitidez na resolução das bandas proteicas dos géis SDS-PAGE. Isso permitirá o mapeamento e busca de novos marcadores da reabsorção radicular externa, tendo em vista a dificuldade de realização de testes moleculares com os atuais candidatos a marcadores proteicos. .
Subject(s)
Adolescent , Adult , Female , Humans , Male , Young Adult , Extracellular Matrix Proteins/analysis , Gingival Crevicular Fluid/chemistry , Root Resorption/metabolism , Biomarkers/analysis , Electrophoresis, Gel, Two-Dimensional/methods , Electrophoresis, Polyacrylamide Gel/methods , Isoelectric Focusing/methods , Phosphoproteins/analysis , Sialoglycoproteins/analysis , Water/chemistryABSTRACT
Cyanosis in an apparently healthy newborn baby may be caused by hemoglobin variants associated with the formation of methemoglobin, collectively known as M hemoglobins. They should not be confused with genetic alterations in methemoglobin reductase enzyme systems of red cells since treatment and prognosis are completely different. A newborn male child was noted to be significantly cyanotic at birth and is the basis for this report. Hemoglobin isoelectric focusing, acid and alkaline gel electrophoresis, and HBA/HBB gene sequencing were performed for the child, both parents and a sister. The newborn child was treated with methylene blue in an intensive care unit fearing that he had a defective reductase system and exposure to oxidant drugs or toxins. Newborn hemoglobin screening with high performance liquid chromatography was abnormal on the 10th and 45th days but no conclusive diagnosis was reached. Cyanosis persisted up to four years of age with no other symptoms. Hemoglobin M Iwate [alpha2 87(F8) His>Tyr, HBA2:c.262C>T] was detected. It was not present in the child's presumed mother, father, sister, and brother. The analysis of 15 short tandem repeats in the trio demonstrated a de novo mutation occurrence (p-value < 1 × 10 -8). The family was reassured that no further action was necessary and genetic counseling was provided. Methemoglobins should be considered for differential diagnosis of cyanosis in newborns even if no familial cases are detected. Except for cosmetic consequences, the clinical course of patients with hemoglobin M Iwate is unremarkable.
Subject(s)
Humans , Male , Female , Hemoglobin A2 , Hemoglobin M , Sequence Analysis, DNA , Cyanosis , Isoelectric Focusing , MethemoglobinemiaABSTRACT
O presente estudo teve como objetivo verificar a eficácia da ferramenta de focalização utilizada em intervenção breve com pacientes em situação de prótese ocular. Foi realizado um estudo quasi-experimental de caso-controle com uma amostra de conveniência, composta por 10 participantes para o grupo experimental e oito para o controle. O Teste de Habilidade Experiencial (THE) foi utilizado para a coleta de dados em dois momentos em desenho de pré-teste/pós-teste. O grupo experimental vivenciou quatro sessões terapêuticas de focalização. O tamanho do efeito da diferença de crescimento da média do grupo experimental e do grupo controle foi pequeno-moderado (d = 0,38; p = 0,44). Implicações dos resultados para a prática da psicologia clínica são discutidas.
The present study aimed to verify the effectiveness of the focusing therapy applied in brief intervention with patients in context of ocular prosthesis. The design of case-control study included a convenience sample with 10 participants for the experimental group and eight participants for the control group. The data was collected by the Experiential Ability Test (EAT) on the pre-test and pos-test. The experimental group experienced four psychotherapy sessions guided by the focusing technique. The effect size of the difference in average growth in the experimental group and control group was small-moderate (d = 0.38, p = 0.44). Implications for the practice of clinical psychology are discussed.
El presente estudio tuvo como objetivo verificar la eficacia de la herramienta de focalización utilizado en breves intervenciones con pacientes con la prótesis ocular. Se realizó un estudio cuasi-experimental de casos y controles con una muestra compuesta por 10 participantes en el grupo experimental y ocho en el de control. El Test de la Habilidade Experiencial (THE) se utilizó para recopilar datos en dos puntos, pré-teste/pós-teste. El grupo experimental experimentaron cuatro sesiones terapéuticas de la focalización. El tamaño del efecto de la diferencia en el crecimiento promedio del grupo experimental y el grupo de control fue pequeña-moderada (d = 0,38, p = 0,44). Se discuten las implicaciones de los resultados para la práctica de la psicología clínica.