Uso de PCR digital para definir terapéuticas específicas y seguimiento en enfermedades autoinflamatorias / Use of digital PCR to define specific therapeutics and follow-up in autoinflammatory diseases

Las enfermedades autoinflamatorias (AIDs) son un grupo heterogéneo de desórdenes monogénicos o poligénicos, con características de disregulación inmune innata y/o adaptativa, cuyo mecanismo central es la autoinflamación pero también pueden presentarse con autoinmunidad e inmunodeficiencia. En estos últimos años el desarrollo de las tecnologías de secuenciación masiva han provocado una explosión en el descubrimiento de nuevos genes responsables de AIDs monogénicas. Esto remarca la importancia de implementar este tipo de estudios para llegar a un diagnóstico definitivo sobre todo en este grupo de patologías genéticamente muy diversas donde los fenotipos clínicos se solapan. Sin embargo, dada la presencia de variantes de significación incierta (VUS), los resultados pueden no ser concluyentes planteándose la necesidad de desarrollar pruebas funcionales para determinar la patogenicidad de dichas variantes genéticas. En nuestro grupo de trabajo estamos aplicando la PCR digital en gotas (ddPCR), una técnica cuantitativa de 3era generación altamente sensible, especifica y reproducible que no necesita de curvas de calibración, para desarrollar pruebas funcionales que permitan no sólo reclasificar variantes VUS para lograr diagnósticos definitivos sino también estudiar los mecanismos responsables de las principales AIDs que permitan una estratificación de las terapéuticas especificas a aplicar y de esta manera poder contribuir al diagnóstico, tratamiento y seguimiento de nuestros pacientes en forma personalizada. (AU)

Autoinflammatory diseases (AIDs) are a heterogeneous group of monogenic or polygenic disorders, with characteristics of inborn and/or adaptive immune dysregulation, whose central mechanism is autoinflammation but may also present with autoimmunity and immunodeficiency. In recent years the development of massive sequencing technologies has led to an exponential increase in the discovery of new genes responsible for monogenic AIDs. This emphasizes the importance of the implementation of this type of studies to make a definitive diagnosis, especially in this group of genetically very diverse diseases with overlapping clinical phenotypes. However, given the presence of variants of uncertain significance (VUS), the results may not be conclusive, raising the need to develop functional tests to determine the pathogenicity of these genetic variants. In our working group we are applying droplet digital PCR (ddPCR), a highly sensitive, specific and reproducible third generation quantitative technique that does not require calibration curves, to develop functional tests that allow not only to reclassify VUS variants to achieve definitive diagnoses but also to study the mechanisms responsible for the main AIDs that allow for the stratification of specific treatments to be used and thereby contribute to the individualized diagnosis, treatment, and follow-up of our patients (AU)

Perfil dos casos de câncer de mama entre acometidos no acre período de 2015 a 2019 ­ um estudo transversal / Profile of breast cancer cases between affected in acre period from 2015 to 2019 - a cross-sectional study / Perfil de los casos de cáncer de mama entre las personas afectadas en el período acre de 2015 a 2019 - un estudio transversal

Este estudo teve como objetivo analisar o perfil dos casos de câncer de mama no estado do Acre no período de 2015 a 2019. Trata-se de um estudo quantitativo com delineamento transversal. A amostra foi constituída por todos os casos de câncer de mama registrados no Acre e inseridos no Departamento de Informática do Sistema Único de Saúde (DATASUS) no período de 2015 a 2019. Foram identificados no período 293 casos da doença, com maior número de casos registrados no ano de 2019 (25,0%), sendo a maioria do sexo feminino (98,0%), na faixa etária de 40 a 49 anos (29,0%). O tempo decorrido desde o diagnóstico até o início do tratamento foi de mais de 60 dias (51,0%). A modalidade terapêutica mais utilizada foi a quimioterapia (55,0%). O local da realização do tratamento ocorreu capital do estado Rio Branco (80,0%). A maior parte dos acometidos, ainda encontra-se em tratamento (56,0%), no entanto (44,0%) evoluiu para óbito. O aumento da doença com o passar dos anos é notável no Acre. É importante destacar que ações voltadas para a prevenção e controle do câncer de mama continuam sendo fundamentais para auxiliar na diminuição do número de casos, como o rastreamento e diagnóstico precoce.

This study aimed to analyze the profile of breast cancer cases in the state of Acre in the period from 2015 to 2019. This is a quantitative study with a cross-sectional design. The sample consisted of all breast cancer cases registered in Acre and inserted in the Informatics Department of the Unified Health System (DATASUS) in the period from 2015 to 2019. In the period 293 cases of the disease were identified, with a greater number of cases registered in 2019 (25.0%), with the majority being female (98.0%), aged 40 to 49 years (29.0%). The time elapsed from diagnosis to the start of treatment was more than 60 days (51.0%). The most used therapeutic modality was chemotherapy (55.0%). The place where the treatment was performed took place in the state of Rio Branco (80.0%). Most of the people affected are still under treatment (56.0%), however (44.0%) died. The increase in the disease over the years is notable in Acre. It is important to highlight that actions aimed at the prevention and control of breast cancer continue to be fundamental to assist in reducing the number of cases, such as screening and early diagnosis.

Este estudio tuvo como objetivo analizar el perfil de los casos de cáncer de mama en el estado de Acre en el período de 2015 a 2019. Se trata de un estudio cuantitativo con diseño transversal. La muestra consistió en todos los casos de cáncer de mama registrados en Acre e ingresados en el Departamento de Informática del Sistema Único de Salud (DATASUS) en el período de 2015 a 2019. En el periodo se identificaron 293 casos de la enfermedad, siendo el mayor número de casos registrados en 2019 (25,0%), siendo la mayoría mujeres (98,0%), en el grupo de edad de 40 a 49 años (29,0%). El tiempo transcurrido desde el diagnóstico hasta el inicio del tratamiento fue superior a 60 días (51,0%). La modalidad terapéutica más utilizada fue la quimioterapia (55,0%). El lugar donde se realizó el tratamiento fue Rio Branco, la capital del estado (80,0%). La mayoría de los pacientes afectados siguen en tratamiento (56,0%), sin embargo, (44,0%) fallecieron. El aumento de la enfermedad a lo largo de los años es notable en Acre. Es importante destacar que las acciones dirigidas a la prevención y control del cáncer de mama siguen siendo fundamentales para ayudar a reducir el número de casos, como el cribado y el diagnóstico precoz.

A network pharmacology approach to assess Reno-protective and -curative effects of methanolic extract of Malva neglecta Wallr in gentamicin induced renal toxicity rat model

Abstract The aim of present study was to explore protective and curative effects of Malve neglecta on kidneys. In silco study with network pharmacology was performed to find out potential target organs, genes and cellular cell lines which confirmed kidneys as target organ of phyto-constituents present in Malva neglecta extract. Gentamicin (40 mg/kg, i.p) was given to induce renal toxicity. Prophylactic study was performed with 300-, 600- and 900 mg/kg doses to find out nephro-protective and -curative effects and curative potential was evaluated at 900 mg/kg dose. Renal function biomarkers, blood urea, BUN, serum creatinine and uric acid, and oxidative stress measuring biomarkers, SOD, CAT, GSH and MDA levels in kidney homogenate were quantified at the end of study. Treatment groups showed decrease in blood urea, BUN, serum creatinine and uric acid levels dose dependently and curative group also showed decline in these biomarkers. SOD, CAT, GSH levels were increased and MDA level decreased in treatment groups significantly as compared to toxic control which revealed the role of oxidative stress in renal damage and anti-oxidant power of MN. Data suggested that use of MN along with drugs causing renal toxicity may prove beneficial due to its nephro- protective and curative effects.

Effects of low-level laser therapy on liver regeneration and laser parameters employed / Efeitos da laserterapia de baixa potência na regeneração hepática e parâmetros empregados de laser

Low-level laser therapy has several biological effects; one of them is tissue regeneration. Recent studies have been held on the application of laser therapy on the liver of rats after partial hepatectomy to promote liver regeneration. The aim of this article was to review the recent studies on the effects of low-level laser therapy on rat liver regeneration after partial hepatectomy and the laser parameters used in those studies. A review of recent relevant literature was performed in Pubmed, Scielo, Medline, and Bireme databases. Articles related to the application of low-level laser therapy on hepatic regeneration were included. Articles with hepatic regeneration in the presence of pathologies were not included. Nine studies were found matching the study criteria. In most studies, low-level laser therapy promoted liver regeneration after partial hepatectomy, without further damage to the remaining liver. Not all laser parameters required for the reproducibility of the study were described by all authors. The therapeutic use of low-level laser therapy in liver regeneration can be promising; however, since the liver is a vital organ, and the laser application is intraoperative, future studies are necessary. The parameters used must be properly described and standardized to allow the reproducibility of the study, in order to define a therapeutic window and thus, consider its clinical use. It is also essential to clarify the mechanisms by which laser promotes liver regeneration to guarantee its safety and therapeutic efficacy.

Laserterapia de baixa potência tem vários efeitos biológicos, sendo um deles a regeneração de tecido. Sua aplicação no fígado de ratos após hepatectomia parcial para promoção de regeneração hepática tem sido estudada recentemente. O objetivo deste artigo foi revisar os estudos recentes dos efeitos da laserterapia de baixa potência na regeneração de fígado de ratos após hepatectomia parcial de fígado e os parâmetros de laser empregados. Uma revisão da literatura relevante recente foi realizada nas bases de dados Pubmed, Scielo, Medline e Bireme. Artigos sobre a aplicação da laserterapia de baixa potência na regeneração de fígado foram incluídos. Artigos sobre regeneração hepática na presença de patologias foram excluídos. Nove estudos foram encontrados correspondendo aos critérios do estudo. Na maioria dos estudos, a laserterapia de baixa potência promoveu regeneração hepática após hepatectomia parcial, sem causar danos adicionais ao fígado remanescente. Não foram descritos todos os parâmetros necessários para reprodutibilidade dos estudos por todos os autores. O uso terapêutico da laserterapia de baixa potência na regeneração de fígado pode ser promissor, entretanto, como o fígado é um órgão vital e a aplicação do laser é intraoperativa, estudos futuros são necessários, assim como os parâmetros da aplicação de laser precisam ser descritos apropriadamente e padronizados, para permitir a reprodutibilidade do estudo, para que uma janela terapêutica possa ser definida e seu uso clínico possa ser considerado. Também é essencial esclarecer através de quais mecanismos o laser promove regeneração de fígado para garantir sua segurança e eficácia terapêutica.

Representaciones de niños y niñas de 8 a 12 años sobre el tratamiento virtual de psicopedagogía durante el contexto de pandemia 2020-2021: Cesac N°15 y Hospital Durand / Representations of children aged 8 to 12 on the virtual treatment of psychopedagogy during the context of pandemic 2020-2021: Cesac N°15 and Hospital Durand

La presente investigación pretende abordar una temática actual surgida en el contexto de pandemia por COVID19 bajo las regulaciones nacionales en relación al aislamiento preventivo y obligatorio vivenciado a lo largo del año 2020. En la misma se indaga sobre las representaciones de los niños y niñas en relación a los tratamientos psicopedagógicos virtuales, entendidos estos como un vínculo estable prolongado en un cierto tiempo entre un niño/a o adolescente (NNA) y un/una profesional de la psicopedagogía con el fin de acompañar o ayudar a disminuir su malestar en torno al aprendizaje en alguna de sus aristas. Estos encuentros desde el inicio de la pandemia se vieron enmarcados en aplicaciones digitales - como zoom o videollamada de Whatsapp debido al contexto de emergencia sanitaria mundial. El objetivo fundamental será investigar las características particulares del tratamiento psicopedagógico bajo la modalidad virtual a partir de las voces de los niños y las niñas que presentan dificultades en sus procesos de aprendizaje, es decir teniendo como eje central a los actores principales del espacio de tratamiento y sus representaciones sobre el mismo. (AU)

Uso da terapia fotodinâmica antimicrobiana no tratamento de candidose oral: ensaio clínico, controlado e randomizado / Use of antimicrobial photodynamic therapy in the treatment of oral candidosis: a randomized, controlled clinical trial

INTRODUÇÃO: A candidose eritematosa oral é uma infecção fúngica comum em usuários de prótese. Em algumas situações os pacientes exibem resistência ao tratamento convencional. Novas terapias vêm surgindo como alternativas para o seu tratamento, como a terapia fotodinâmica antimicrobiana (TFDa). OBJETIVO: Analisar a eficácia da TDFa no tratamento da candidose eritematosa oral em comparação com o tratamento convencional a base de nistatina. METODOLOGIA: O presente estudo consiste em um ensaio clínico, controlado e randomizado, realizado por meio de um estudo clínico e análise microbiológica em pacientes diagnosticados com candidose eritematosa oral, os quais foram divididos entre grupo controle, tratados com suspensão oral de nistatina e grupo caso, tratados com sessões de TFDa. Os pacientes foram avaliados quanto a eficácia do tratamento, através da aplicação de um índice clínico, além de serem realizadas análises microbiológicas, em períodos préestabelecidos (7,14,21 e 30 dias). RESULTADOS: Dos 41 pacientes analisados, sendo 18 deles do grupo controle e 23 do grupo caso, 32 (78%) obtiveram sucesso dos tratamentos utilizados. Quanto ao grau clínico, as severas exibiram mais dificuldade em apresentar remissão, enquanto todas as lesões leves e moderadas regrediram. O grupo caso, exibiu resultados semelhantes ao do grupo controle, no entanto, não houve diferenças estatisticamente significativa entre os grupos, tanto quanto ao sucesso do tratamento, nas análises microbiológicas, no qual foi contatado o predomínio da Candida albicans,entre as espécies de Candida ssp. e de Staphylococcus ssp. nas próteses dentarias. CONCLUSÕES: A TFDa é uma alternativa de tratamento promissora, por apresentar resultados satisfatórios, não causar alterações aos tecidos orais e seu uso não levar ao desenvolvimento de resistência ao tratamento (AU).

INTRODUCTION: Oral erythematous candidiasis is a common fungal infection in users of complete dentures. patients often exhibit resistance to conventional treatment. New therapies are emerging as alternatives for its treatment, such as antimicrobial photodynamic therapy(aPDT). AIM:Analysis in patients diagnosed with oral erythematous candidiasis, who were divided into a control group, treated with oral nystatin suspension, and an intervention group, treated with aPDT sections. METHODS: The present study consists of a clinical, controlled and randomized trial conducted by means of a clinical and microbiological analysis in patients diagnosed with oral erythematous candidiasis, who were divided into a control group, treated with oral nystatin suspension, and an intervention group, treated with aPDT sections. Patients were reassessed as to the effectiveness of treatment through the application of a clinical index, in addition to microbiological analysis,in pre-established periods (7,14,21 and 30 days).RESULTS:Of 41 patients analyzed in our research, 32 (78%) were successful in their treatment,4 (9,8%) were unsuccessful, and 5 (12,2%) gave up treatment. Patients were also analyzed for the degree of the lesion, and it was observed that those of the severe type had more difficulty in presenting remission, while the lesions mild and moderate, all showed regression. The case group exhibited similar results to the control group, however, there were no statistically significant differences between the groups, as far as treatment success was concerned, in microbiological analyzes, in which the predominance of Candida albicans, among the species of Candida ssp. and Staphylococcus ssp. in dental prostheses. CONCLUSIONS: aPDT is a promising treatment alternative, as it presents satisfactory results, does not cause changes to oral tissues and its use does not lead to the development of resistance to treatment, so further studies with larger samples are needed to verify its effectiveness (AU).

Investigación 2019-2020: Razones prevalentes de la asistencia regular al tratamiento psicopedagógico y grado de satisfacción con el mismo, en el Área Programática del Hospital Gral. de Agudos Dr. J. M. Ramos Mejía: ¿Por qué sostienen los que sostienen?" / Research 2019-2020: Prevalent reasons for the regular attendance to the psychopedagogical treatment and degree of satisfaction, in the Programmatic Area of the Hospital Gral. Dr. J. M. Ramos Mejía: Why do you hold those who hold?

Investigación del equipo de psicopedagogas residentes del Área Programática del Hospital Ramos Mejía (HRM), de la Ciudad de Buenos Aires, donde la clínica psicopedagógica es una de las principales tareas que desempeña, destinada a una población de niños/as entre 4 y 11 años. Desde 2019, se distribuye la encuesta de seguimiento por tratamiento sostenido, instrumento que se administró a los referentes de los niños/as que habían podido sostener el tratamiento. Considerando la multiplicidad de factores que pudieran afectar la asistencia regular, dicha encuesta incluye preguntas que indagan el motivo de llegada al equipo, la presencia de preocupaciones al iniciar el proceso, el tiempo de tratamiento, las razones explicitadas por las que se sostiene el espacio, la conveniencia del turno en cuanto día y horario, el tiempo de traslado al Área Programática, la calidad de la comunicación con la psicopedagoga del niño/a, entre otras. En el presente trabajo, a partir de la información antes mencionada, se procura analizar las razones prevalentes para el sostenimiento de los tratamientos psicopedagógicos en la población del área y su grado de satisfacción con el mismo.

Skin-Mountable Biosensors and Therapeutics: A Review.

Miniaturization of electronic components and advances in flexible and stretchable materials have stimulated the development of wearable health care systems that can reflect and monitor personal health status by health care professionals. New skin-mountable devices that offer seamless contact onto the human skin, even under large deformations by natural motions of the wearer, provide a route for both high-fidelity monitoring and patient-controlled therapy. This article provides an overview of several important aspects of skin-mountable devices and their applications in many medical settings and clinical practices. We comprehensively describe various transdermal sensors and therapeutic systems that are capable of detecting physical, electrophysiological, and electrochemical responses and/or providing electrical and thermal therapies and drug delivery services, and we discuss the current challenges, opportunities, and future perspectives in the field. Finally, we present ways to protect the embedded electronic components of skin-mountable devices from the environment by use of mechanically soft packaging materials.

The Effect of Medical Technology Innovations on Patient Outcomes, 1990-2015: Results of a Physician Survey.

BACKGROUND: Developments in diagnostics, medical devices, procedures, and prescription drugs have increased life expectancy and quality of life after diagnosis for many diseases. Previous research has shown that, overall, increased investment in medical technology has led to increased health outcomes. In addition, the value of investment in specific innovations, particularly in new pharmaceuticals or biopharmaceuticals, has frequently been shown through an evaluation of the associated health outcomes and costs. Value assessments for all medical technologies and interventions are an important consideration in current debates on access and affordability of health care in the United States. OBJECTIVE: To identify practicing physician impressions of the historical effect of postdiagnosis innovations in medical technology on patient outcomes within the 8 health conditions that have the largest effect on health in the United States. METHODS: National statistics were used to identify the 8 conditions responsible for the most mortality and morbidity within the United States between 1990 and 2014. A physician survey was developed for each major condition to obtain physician opinion on the extent to which pharmaceuticals and biopharmaceuticals, medical devices, diagnostics, and surgical procedures contributed to improvements in postdiagnosis mortality and morbidity outcomes over the evaluated period. Respondents were provided with a fifth category, "cannot allocate," to account for postdiagnosis outcome gains resulting from other factors such as public health interventions. RESULTS: The conditions identified as having the greatest effect on morbidity and mortality since 1990 were breast cancer, ischemic heart disease, human immunodeficiency virus infection, diabetes, unipolar depression, chronic obstructive pulmonary disease, cerebrovascular disease, and lung cancer. After excluding other factors, physicians specializing in these conditions, with a mean of 21.4 years in practice, considered pharmaceuticals and biopharmaceuticals as having the greatest postdiagnosis effect across all 8 conditions, with 56% of outcome gains attributed to this innovation category. Diagnostics was the second biggest contributor at 20%. CONCLUSIONS: Physician perceptions indicated that attention should be paid to value assessments of innovative diagnostics, devices, and surgical procedures, as well as to pharmaceuticals and biopharmaceuticals, before goals for allocating health care expenditures among the different innovations are determined. DISCLOSURES: Funding for this study was provided by the National Pharmaceutical Council, a health policy research group that receives its funding from biopharmaceutical manufacturers. Wamble is employed by RTI Health Solutions, which received funding from the National Pharmaceutical Council to conduct this research. Ciarametaro and Dubois are employed by the National Pharmaceutical Council.

Evaluation of the IncoStress device for urinary incontinence: a feasibility study and pilot randomised controlled trial.

INTRODUCTION AND HYPOTHESIS: The aim of this study was to assess the feasibility of recruitment to and outcomes from a pilot randomised study of the IncoStress device as an adjunct to conservative treatment for urinary incontinence. METHODS: Women with urinary incontinence were randomised on a 2:1 basis to usual care (control) or usual care plus use of the IncoStress device (intervention). Process outcomes (retention and compliance) were recorded plus symptom outcomes (IQOL and ICIQ-FLUTS questionnaires). A sample of participants took part in an interview to understand the frequency of use of the device and satisfaction. RESULTS: Eighty women (51 intervention, 29 control) were recruited. Follow-up responses were obtained from 34 intervention group (66.7%) and 17 (58.6%) control patients. Women used the device for a median 3 days a week (0-7), 18 out of 34 (53%) found it easy to use and 21 (61.8%) were satisfied with the device. Median IQOL score in the intervention group improved from a baseline of 42.4 (0-94) to 68.2 (5-98) at follow-up and in the control group from 45.5 (0-88) to 53.0 (0-94). Median ICIQ-FLUTS score in the intervention group improved from 14.5 (6-35) to 12.5 (4-26) and in the control group from 15.0 (5-35) to 14.0 (6-38). CONCLUSIONS: Recruitment and randomisation were feasible and robust. This study demonstrates that a large-scale RCT is feasible and the IncoStress has potential value.

Facial asymmetry correction with moulded helmet therapy in infants with deformational skull base plagiocephaly.

PURPOSE: The recommendation issued by the American Academy of Pediatrics in the early 1990s to position infants on their back during sleep to prevent sudden infant death syndrome (SIDS) has dramatically reduced the number of deaths due to SIDS but has also markedly increased the prevalence of positional skull deformation in infants. Deformation of the base of the skull occurs predominantly in very severe deformational plagiocephaly and is accompanied by facial asymmetry, as well as an altered ear position, called ear shift. Moulded helmet therapy has become an accepted treatment strategy for infants with deformational plagiocephaly. The aim of this study was to determine whether facial asymmetry could be corrected by moulded helmet therapy. MATERIALS AND METHODS: In this retrospective, single-centre study, we analysed facial asymmetry of 71 infants with severe deformational plagiocephaly with or without deformational brachycephaly who were undergoing moulded helmet therapy between 2009 and 2013. Computer-assisted, three-dimensional, soft-tissue photographic scanning was used to record the head shape before and after moulded helmet therapy. The distance between two landmarks in the midline of the face (i.e., root of the nose and nasal septum) and the right and left tragus were measured on computer-generated indirect and objective 3D photogrammetry images. A quotient was calculated between the two right- and left-sided distances to the midline. Quotients were compared before and after moulded helmet therapy. Infants without any therapy served as a control group. RESULTS: The median age of the infants before onset of moulded helmet therapy was 5 months (range 3-16 months). The median duration of moulded helmet therapy was 5 months (range 1-16 months). Comparison of the pre- and post-treatment quotients of the left vs. right distances measured between the tragus and root of the nose (n = 71) and nasal septum (n = 71) revealed a significant reduction of the asymmetry (Tragus-Nasion-Line Quotient: 0.045-0.022; p < 0.0001; Tragus-Subnasale-Line Quotient: 0.045-0.021; p < 0.0001). The control group without treatment showed no significant change in the quotient (Tragus-Nasion-Line Quotient no helmet: 0.049-0.055/Tragus-Subnasale-Line Quotient no helmet: 0.039-0.055). CONCLUSION: Moulded helmet therapy can correct facial symmetry in infants with deformational plagiocephaly and associated facial and basal skull asymmetry.

Intravenous immunoglobulin therapy among pediatric patients: labeled and off-labeled indications

ABSTRACT This study was designed to evaluate utilization patterns and clinical outcome of intravenous immunoglobulin (IVIG) therapy among pediatric patients in a tertiary hospital. Demographic data, IVIG prescribed, and clinical outcome were retrospectively reviewed from the pharmacy dispensing data and patient medical records between 2007 and 2014. One hundred and fifteen instances of IVIG administration to 108 pediatric patients were recorded. A total of 61 cases (53%) and 54 cases (47%) of the IVIG administered were for labeled and off-labeled indications, respectively. Age, weight, specialty, total IVIG usage, length of hospital stays, and mortality rate were found to be significantly associated with the indication being labeled or off-labeled (p<0.05). However, there was no significant difference in terms of adverse reactions between labeled and off-labeled indications (p>0.05). Guidelines should be developed and implemented for rational and evidence-based use of IVIG to avoid unnecessary wastage.

Avaliação de terapias imunossupressoras em transplantes renais com uma abordagem metabolômica / Assessment of immunosuppressive therapies with a metabolomic approach

O aprimoramento das técnicas analíticas viabilizou a metabolômica, uma área da ciência que busca compreender, de forma comparativa, os metabólitos envolvidos nas vias bioquímicas. A metabolômica está inserida no contexto das "ômicas", que teve início na "Era Genômica", a qual permitiu a identificação de diversos genes. Em seguida, o interesse dos pesquisadores centrou no estudo dos metabólitos (metabolômica) mostrando ser uma ferramenta valiosa na pesquisa do transplante renal, que exige um tratamento medicamentoso por meio de imunossupressores. A combinação destes imunossupressores pode minimizar a rejeição do órgão transplantado, reforçando a necessidade de um estudo metabolômico, a fim de avaliar e comparar as mudanças ocorridas após o transplante em nível molecular, melhorando o conhecimento sobre a influência destes regimes e dando subsídios sobre prognósticos possíveis na área de transplante renal. Nesta tese foram avaliadas 2 terapias: Everolimo/ Prednisona/Tacrolimo (grupo 1) e Micofenolato mofetil/Prednisona/Tacrolimo (grupo 2) a partir de uma abordagem untargeted. No presente trabalho foram coletadas amostras de urina de pacientes ao longo de 6 meses. Foi necessário determinar a melhor condição para análise das amostras de urina dos pacientes. Desta forma, foram realizados estudos sobre alguns parâmetros que impactam no preparo de amostra abordando a influência da urease, tipos e proporção de solventes para precipitação de proteína, seleção do melhor agente derivatizante e tratamento de dados. A avaliação da medida de qualidade dos tratamentos com urease foi feita a partir do desvio padrão relativo (RSD) dos valores de intensidade de pico. A concentração de 10 mg mL-1 apresentou o melhor resultado. O estudo mostrou também que o teor de ureia na urina pode influenciar na identificação dos compostos. O número de compostos identificados foi menor quando a urina não foi tratada com urease, com aproximadamente 10 compostos a menos em relação à amostra tratada com a enzima, na mesma concentração de ureia adicionada. Dos solventes orgânicos testados para precipitação de proteínas nas amostras de urina, o isopropanol mostrou ser o solvente mais adequado na proporção 1:6 urina:solvente (v/v), utilizando-se 100 µL de urina. Foram testados dois protocolos de derivatização para análises por GC-MS: metoximação e sililação utilizando BSTFA e cloroformiato de metila. A comparação mostrou que o procedimento por BSTFA, com 40 metabólitos identificados, foi superior ao cloroformiato de metila, com 13 compostos identificados. No tratamento de dados com o software XCMS, os seguintes parâmetros foram avaliados: largura a meia altura do pico (fwhm), largura da banda (bw) e threshold (sntresh). Para avaliar a melhor combinação destes parâmetros, foi feita uma variação univariada destes valores. A qualidade do resultado de cada combinação foi monitorada pelos valores gerados de número de missing values, quantidade de picos com RSD <15% e número de valores duplicados. Os valores ótimos foram obtidos para a combinação: fwhm=4, bw=2 e threshold=5. A abordagem do estudo dos dois grupos de pacientes baseou-se inicialmente na comparação entre o dia 7 da terapia com os demais períodos (dia 14, mês 1, mês 3 e mês 6) e posteriormente avaliou-se a evolução temporal. A partir do mês 3 os valores de correlação e predição dos modelos de PLS-DA são melhores e já é eficaz na diferenciação entre os dois grupos. Foram observadas perturbações no metabolismo de carboidratos em ambos os grupos, como açúcares, glicerol e N-acetil-D-manosamina. No grupo 1, foram observados metabólitos discriminantes da classe dos poliois e das vias do ciclo do ácido cítrico e degradação de xenobióticos, enquanto que, no grupo 2, foi observada alteração do hidroxibutirato, um corpo cetônico. Neste grupo, foi observado também um aumento do ácido hipúrico, ácido acetamido butírico, ácido benzoico, entre outros. Nesta tese foi possível demonstrar que a metabolômica é uma ferramenta importante para comparar metabólitos discriminantes entre dois regimes imunossupressores, sendo um estudo piloto que visa fornecer subsídios para avaliação da influência destas terapias no prognóstico de transplante renal

The improvement of analytical techniques enabled the emergence of metabolomics, which aims to compare the metabolites involved in biochemical pathways, in certain biological conditions. Metabolomics is inserted in the "omics" context, which began in the "Genomic Age", and allowed the identification of several genes. After that, the researchers focused on the study of metabolites. Among several applications, metabolomics can be a valuable tool in renal transplant research, which requires a drug treatment through immunosuppressants. The combination of these immunosuppressants can minimize toxicity and rejection of the transplanted organ, reinforcing the need for a metabolomic study, in order to evaluate and compare changes after transplantation at the molecular level, improving knowledge about the influence of these regimens and giving subsidies on prognosis in the area of renal transplantation. In this thesis two immunosuppressive therapies were evaluated by an untargeted approach: Everolimus/Prednisone/Tacrolimus (group 1) and Mycophenolate mofetil/Prednisone/Tacrolimus (group 2). In this study, urine samples were collected from patients over 6 months. It was necessary to determine the best condition for analysis of patients' urine samples. Thus, studies were carried out on some parameters that impact on sample preparation, evaluating the influence of urease, types and proportion of solvents for protein precipitation, selection of the best derivatizing agent, and data treatment. The evaluation of the quality measure of the urease treatments was made from the relative standard deviation (RSD) of the peak intensity values. The concentration of 10 mg mL-1 presented the best result. The study also showed that urine urea content may influence the identification of the compounds. The number of identified compounds was lower when urine was not treated with urease, with approximately 10 compounds less than the enzyme-treated sample, at the same concentration of urea added. In the evaluation of the organic solvents tested for protein precipitation in the urine samples, isopropanol was the most suitable solvent in the ratio 1: 6 urine:solvent (v/v), using 100 µL of urine. Two derivatization protocols were tested for GC-MS analysis: metoximation and silylation with BSTFA and methyl chloroformate. The comparison between the two derivatization protocols showed that the BSTFA procedure, with 40 identified metabolites, was superior to methyl chloroformate with 13 compounds identified. In data processing with the XCMS software, the following parameters were evaluated: full width at half maximum of the peak (fwhm), bandwidth (bw) and threshold (sntresh). To evaluate the best combination of these parameters, a univariate variation of these values was made. The quality of the result of each combination was monitored by the number of missing values, number of peaks with RSD <15%, and number of duplicate values. The optimal values were obtained for the combination: fwhm=4, bw=2 and threshold =5. The study of the two groups of patients was initially based on the comparison between day 7 of the therapy with the other periods (day 14, month 1, month 3 and month 6) and later the temporal evolution was evaluated. From month 3 the values of correlation and prediction of the PLS-DA models are better and already effective in the differentiation between the two groups. Disorders in carbohydrate metabolism were observed in both groups with sugars and glycerol and N-acetyl-D-mannosamine as discriminant metabolites. In group 1, discriminant metabolites of the class of polyols and citric acid cycle pathways and degradation of xenobiotics were observed, and in group 2 alteration of hydroxybutyrate, a ketone body, was observed. In this group an increase of hippuric acid, acetamido butyric acid, benzoic acid, among others, was also observed. In this thesis it was possible to demonstrate that metabolomics is an important tool to compare discriminant metabolites between two immunosuppressive regimens, being a pilot study that aims to provide future subsidies to evaluate the influence of these therapies on the renal transplant prognosis

The effect of continuous elastic outside distraction on the expressions of vascular endothelial cell growth factor and microvessel density in female porcine nipple.

BACKGROUND: We have proposed a method to correct the inverted nipple with good clinical results. The aim of the study presented here is to show the effect of continuous elastic distraction on the expression of vascular endothelial growth factor and microvessel density in the female porcine nipple. This article also explores the probable mechanism correction of inverted nipples. METHODS: Four female 3-month-old pigs each with 12 nipples were used. Four nipples of each minipig were used as control with the other eight nipples continuously distracted with prefabricated instruments. The nipples were excised at weeks 2, 4, 8, and 12 after distraction. Immunohistochemical staining was performed to observe the expression of vascular endothelial cell growth factor (VEGF) and CD34 in those tissues. The integral optical density of VEGF-positive cells and microvessel density (MVD) were also counted. RESULTS: The volumes of the nipples all increased after traction compared with those of the control group. The diameters and heights of the nipples showed significant changes (P<0.01). In immunohistochemical staining, the expressions of VEGF and CD34 of all distracted groups were positive. The staining intensity of traction groups was moderate and that of the control groups was weakly positive. The integral optical density of VEGF-positive cells and MVD compared with the control group were significantly different (P<0.01), and there was a significant positive correlation in MVD and VEGF. CONCLUSION: Continuous elastic distraction can promote the expression of some growth factors in nipples.

Quiste subcondral a nivel de la articulación interfalángicaproximal en una potranca criolla colombiana: reporte de caso / Subchondral cyst in the proximal interphalangeal joint in a paso fino colombian horse: a case report / Cisto subcondral na articulação proximal interfalângicaem um potro crioulo colombiano: relato de caso

Se reporta el caso de una paciente equina, evaluada por Especial•Vet práctica privada, la cual presentaba alexamen clínico ortopédico un grado de claudicación II/V en ambos miembros posteriores (según clasificaciónde la AAEP), la cual no presento mejoría después de realizar un tratamiento médico de tipo parenteral confenilbutazona. Posteriormente se realizó un nuevo examen clínico ortopédico en el cual se realizó bloqueoanestésico perineural abaxial en ambos miembros posteriores encontrando una mejoría del 90% con respectoal grado de claudicación inicial. Se realizó evaluación radiológica digital, con las siguientes proyecciones:dorso plantar y lateromedial en las cuales se evidenció un área radiolúcida circunscrita a nivel del terciodistal de la primera falange, con comunicación a la articulación interfalángica proximal en ambos miembros posteriores, seguidamente se realizó evaluación ultrasonográfica en la cual se observa un área anecóica y lafalta de continuidad de la superficie ósea a nivel de la articulación interfalángica proximal de ambos miembrosposteriores. Estableciendo de esta forma como diagnóstico definitivo quiste subcondral a nivel del tercio distalde la primera falange, con comunicación a la articulación interfalángica proximal. Se realizó infiltración conacetato de triamcinolona, betametasona y ácido hialurónico a nivel intrarticular; antibioterapia de maneraprofiláctica al procedimiento, descanso en pesebrera por 4 semanas y reincorporación al ejercicio de maneraprogresiva, suministro de complementos condroprotectores de manera enteral (Flexequin® 40 gr/día VO yCortaflex® 20 ml/día VO). Al momento de la publicación de este artículo, la paciente no presenta ningún gradode claudicación y se encuentra realizando un trabajo físico y atlético normal.

We report the case of an equine patient, assessed by Especial Vet private practice, whose orthopedic clinicalexamination showed a degree of lameness II / V in both hind limbs (according to AAEP classification), whichdemonstrated no improvement after medical treatment with parenteral phenylbutazone. Subsequently a neworthopedic clinical examination was performed in which an abaxial, perineural anesthetic block was applied to both hind legs, which produced 90% improvement compared to the initial degree of lameness. Digital radiographicevaluation was performed with the following results: dorsal-plantar and lateral-medial images which showeda circumscribed, radiolucent area at the level of the distal third of the first phalanx, with communication to theproximal interphalangeal joint on both hind limbs. Following, an ultrasound evaluation was carried out in whichthere was an anechoic area and lack of continuity of the bone surface at the proximal interphalangeal joint of bothhind limbs. These findings established a definitive diagnosis of a subchondral bone cyst at the distal third of the firstphalanx, with communication to the proximal interphalangeal joint. Intra-articular infiltration was performed with triamcinolone acetonide, betamethasone and hyaluronic acid; antibiotics as prophylaxis, rest in a stable for 4 weekswith a gradual return to exercise, and provision of enteral, chondroprotective supplements (p.o. Flexequin ® 40 gr/day and p.o. Cortaflex ® 20ml/day). At the time of publication of this article, the patient does not present any degreeof lameness and is performing normal athletic and physical activity.

Relatamos um caso de um paciente eqüino, avaliado pela prática privada Especial Vet, cujo exame clínicoortopédico mostrou um grau de claudicação II / V em ambos os membros posteriores (de acordo com aclassificação do AAEP), o qual não demonstrou melhora após o tratamento médico com fenilbutazona parenteral.Após a aplicação de um bloqueio anestésico perineural abaxial em ambas as pernas traseiras, foi realizadoum novo exame clínico ortopédico, mostrando uma melhora de 90% em comparação com o grau inicial declaudicação. Realizou-se também uma avaliação radiográfica digital obtendo-se os seguintes resultados: imagensdorso-plantar e latero-medial que mostrou uma área radiolúcida circunscrita ao nível do terço distal da primeirafalange, com comunicação para a articulação interfalângica proximal em ambos os membros posteriores. E após,a realização de um ultra-som, verificou-se que houve uma área anecóica e falta de continuidade da superfície óssea ao nível da articulação interfalângica proximal dos dois membros posteriores. Desta forma estabeleceu umdiagnóstico definitivo de um cisto ósseo subcondral no terço distal da primeira falange, com comunicação para aarticulação interfalângica proximal. Uma infiltração intra-articular foi realizada com acetato de triamcinolona,betametasona e ácido hialurônico; antibióticos como profilaxia, um descanso em estábulo durante 4 semanas, com um retorno gradual aos exercícios, e administração de suplementos condroprotetores de maneira enteral(Flexequin® 40 gr/día VO e Cortaflex® 20 ml/día VO). No momento da publicação deste artigo, o paciente nãoapresenta qualquer grau de claudicação e está realizando atividades atléticas e físicas normais.

Therapeutic apheresis instrumentation.

This article provides a concise overview of therapeutic apheresis medicine instrumentation. Three instrument modalities are discussed: (1) centrifugation-based instruments, (2) extracorporeal photopheresis instruments (a subset of centrifugation-based instruments), and (3) column-based instruments.

Nanotechnology and nuclear medicine; research and preclinical applications.

The birth of nanotechnology in human society was around 2000 years ago and soon found applications in various fields. In this article, we highlight the current status of research and preclinical applications and also future prospects of nanotechnology in medicine and in nuclear medicine. The most important field is cancer. A regular nanotechnology training program for nuclear medicine physicians may be welcome.