RESUMO
This study investigated if metabolomic profiling of culture media using near infrared (NIR) spectroscopy was related to live-birth rates after single-embryo transfer of frozen-thawed embryos. Analysis of culture media of frozen-thawed embryos was performed by NIR spectroscopy. A viability score was calculated using a predictive multivariate algorithm of fresh day-5 embryos with known pregnancy outcomes. This algorithm generated with fresh day-5 embryos could help to identify the live-birth group from the no live-birth group. Multivariable regression models that tested the predictive ability of the viability score for live birth showed an odds ratio in the crude analysis of 1.50 (P=0.008), after adjustment for embryo morphology, 1.44 (P=0.022), and after adjustment for all variables, 1.71 (P=0.005); based on a 0.1 step increase in viability scores. In conclusion, higher viability scores resulted in higher live-birth rates. An algorithm generated from fresh embryos might be used to predict viability of frozen-thawed embryos. Frozen-thawed embryos have different metabolic activity which is related to implantation potential. Therefore, this method might be useful to select the best embryo for transfer within a group of embryos with similar morphology.
Assuntos
Blastocisto/metabolismo , Meios de Cultura/química , Metabolômica/métodos , Adulto , Algoritmos , Criopreservação , Técnicas de Cultura Embrionária , Feminino , Humanos , Análise Multivariada , Gravidez , Resultado da Gravidez , Análise de Regressão , Transferência de Embrião Único , Espectroscopia de Luz Próxima ao Infravermelho/métodosRESUMO
PURPOSE: Assessment of embryo viability is a key component of in vitro fertilization (IVF) and currently relies largely on embryo morphology and cleavage rate. In this study, we used receiver operating characteristic (ROC) analysis to compare the Viability Score (generated by metabolomic profiling of spent embryo culture media using near infrared (NIR) spectroscopy) to morphologic grading for predicting pregnancy in women undergoing single embryo transfer (SET) on day 5. METHODS: A total of 198 spent embryo culture media samples were collected in four IVF centers located in the USA, Europe and Australia. First, 137 samples (training set) were analyzed by NIR to develop an algorithm that generates a Viability Score predictive of pregnancy for each sample. Next, 61 samples (validation set) were analyzed by observers blinded to embryo morphology and IVF outcome, using the Day 5 algorithm generated with the training set. Pregnancy was defined as fetal cardiac activity (FCA) at 12 weeks of gestation. RESULTS: The Area Under the Curve (AUC) was greater for the metabolomic Viability Score compared to Morphology [Training set: 0.75 versus 0.55, p = 0.0011; Validation set: 0.68 versus 0.50, P = 0.021], and for a Composite score (obtained using a model combining Viability Score with morphologic grading), compared to morphology alone [0.74 versus 0.50, p = 0.004]. CONCLUSIONS: Our findings suggest that Viability Score alone or in combination with morphologic grading has the potential to be a better classifier for pregnancy outcome than morphology alone in women undergoing SET on day 5.
Assuntos
Implantação do Embrião/fisiologia , Embrião de Mamíferos/anatomia & histologia , Fertilização in vitro , Metabolômica/estatística & dados numéricos , Curva ROC , Desenvolvimento Embrionário , Feminino , Humanos , Gravidez , Prognóstico , Transferência de Embrião ÚnicoRESUMO
The aim of the current study was to evaluate whether near-infrared (NIR) spectroscopy-generated metabolomic data obtained from oocyte culture samples would correlate with nuclear maturity status and derived embryo development. A total of 412 oocyte culture samples were collected from 43 patient cycles. Metabolomic profiles of metaphase I and II oocytes were obtained by NIR spectroscopy and were significantly different from each other and from profiles of prophase I (germinal vesicle) oocytes (P +/- 0.001 at the 95% confidence interval). Additionally, NIR spectroscopic analysis of culture medium of oocytes that developed to grade A embryos on day 3 demonstrated significantly higher viability indices (0.62 +/- 0.23) than those that developed to grades C/D (0.42 +/- 0.26; P < 0.006); and on day 5 grade A (0.37 +/- 0.20) was also higher than grades C/D (0.14 +/- 0.21; P < 0.02). Metabolomic profiles of oocytes that resulted in pregnancy had higher viability indices (0.87 +/- 0.27) than those that did not (0.44 +/- 0.17; P < 0.0001). The results of the current study demonstrate that metabolomic profiling from spent culture medium of the oocyte is related to nuclear maturity, is able to predict embryo development at day 3 and day 5 stages, and relates to embryo viability.
Assuntos
Metabolômica/métodos , Oócitos/metabolismo , Oócitos/fisiologia , Adulto , Biomarcadores/análise , Biomarcadores/metabolismo , Sobrevivência Celular/fisiologia , Células Cultivadas , Meios de Cultura/química , Transferência Embrionária , Feminino , Humanos , Masculino , Oócitos/citologia , Gravidez , Taxa de Gravidez , Controle de Qualidade , Projetos de Pesquisa , Sensibilidade e Especificidade , Espectroscopia de Luz Próxima ao Infravermelho , Injeções de Esperma Intracitoplásmicas/métodosRESUMO
Bisphosphonate-related osteonecrosis of the jaw (BRONJ) was first mentioned in the literature in 2003. Since then, several reports have been published referring to this disease. The etiology of BRONJ still remains unclear. The treatment of BRONJ also remains a topic of discussion between those who are in favor of a conservative treatment and those who are convinced that surgical treatment gives the best results. In this case report, a patient is presented with BRONJ in the mandible which has been treated surgically in combination with antibiotic treatment. During surgery it appeared that a large part of the jaw was sequestrated full-thickness with, at the same time, formation of a substantial amount of subperiosteal bone that was formed around the BRONJ, supporting the sequestrated part of the mandible and, after sequestrectomy, serving as a neo-mandible. This case shows the capacity of the jawbone despite bisphosphonate use to regenerate itself.
RESUMO
AIM: There are no established chemical biomarkers of idiopathic Parkinson's disease (PD). The results of a prior metabolomics-based biomarker study suggested that near-infrared spectroscopy of blood plasma samples may distinguish idiopathic PD from neurologically normal controls. METHODS: Near-infrared spectroscopy was used to detect and quantify substrate modifications in blood plasma samples derived from 71 PD subjects enrolled in the PostCEPT observational study and 68 normal control subjects. RESULTS: Near-infrared spectra values were significantly higher in the PD group compared with the control group when adjusted for age and gender (PD: adjusted mean 0.49 [95% CI: 0.45-0.53]; control: adjusted mean 0.40 [95% CI: 0.36-0.44]; p = 0.004, multiple regression). CONCLUSION: Near-infrared spectra of blood plasma in early-stage idiopathic PD differ from those of non-neurological control subjects. However, the degree of overlap between the groups limits the use of this technique in its present form as a PD biomarker for routine clinical practice.
Assuntos
Biomarcadores/sangue , Doença de Parkinson/sangue , Doença de Parkinson/diagnóstico , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Idoso , Distribuição de Qui-Quadrado , Feminino , Humanos , Masculino , Metabolômica/métodos , Pessoa de Meia-Idade , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
OBJECTIVE: To determine whether metabolomic profiling of spent embryo culture media correlates with reproductive potential of human embryos. DESIGN: Retrospective study. SETTING: Academic and a private assisted reproductive technology (ART) programs. PATIENT(S): Women undergoing single embryo transfer after IVF. INTERVENTION(S): Spent embryo culture media were collected after single embryo transfer on day 3 (n = 304) or day 2 (n = 181) and analyzed by near infrared spectroscopy. Near infrared spectral regions were correlated to reproductive potential using a genetic algorithm optimization. Models of these spectral regions were used to calculate viability indices, and were validated by blinded analysis of a subset (n = 60) of samples. Implantation rates were also compared between embryos of higher (>or=0.3) and lower (<0.3) viability indices, and within each morphology grade. MAIN OUTCOME MEASURE(S): Viability index and embryo viability. RESULT(S): Mean viability indices of embryos that resulted in positive fetal cardiac activity were significantly higher compared with embryos that did not for both day 2 and day 3 embryos. Blinded validation of the day 2 model proved to be significant. Increasing viability index values correlated with an increase in pregnancy. Viability indices were found to be independent of morphology for both day 2 and day 3 embryos. Implantation rates were significantly higher among embryos with viability indices >or=0.3. CONCLUSION(S): Metabolomic profiling of human embryo culture media using near infrared spectroscopy is independent of morphology and correlates with reproductive potential of embryos.
Assuntos
Blastocisto/metabolismo , Meios de Cultura/metabolismo , Metaboloma , Resultado da Gravidez , Transferência de Embrião Único/métodos , Algoritmos , Blastocisto/citologia , Sobrevivência Celular , Técnicas de Cultura Embrionária/métodos , Técnicas de Cultura Embrionária/normas , Feminino , Fertilização in vitro/métodos , Fertilização in vitro/normas , Humanos , Modelos Biológicos , Valor Preditivo dos Testes , Gravidez , Reprodutibilidade dos Testes , Estudos Retrospectivos , Transferência de Embrião Único/normas , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Espectroscopia de Luz Próxima ao Infravermelho/normasRESUMO
The biotechnology industry has undergone rapid growth in recent years largely due to the development and success of protein-based therapeutics for a wide range of disorders. Similar to traditional pharmaceuticals, characterization of a therapeutic protein for its physicochemical properties, process monitoring and lot release is crucial. Electrophoresis in the slab-gel format has and continues to be a mainstay of the protein laboratory; and more recently, CE has begun to make significant inroads for protein analysis in industrial settings. This review focuses on the electrophoresis of proteins with an emphasis on protein-based therapeutics in the capillary, slab-gel and to a lesser extent, the microchip format. Reported applications of electrophoresis at several stages of the biopharmaceutical industry covering the period of 2000-2005 will be discussed.
Assuntos
Eletroforese Capilar/métodos , Eletroforese em Microchip/métodos , Preparações Farmacêuticas/análise , Proteínas Recombinantes/análise , BiotecnologiaRESUMO
Recently it was demonstrated that a collimated Gaussian beam can be converted to a flattop beam using a Keplerian beam reshaper consisting of two aspheric lenses. Here, using the same optical system, we demonstrate that this flattop profile can be maintained when used in a confocal detection system that focuses the laser beam into a diameter of only 33 microm. The intensity profile of the reshaped beam was determined by imaging the excitation of a constant stream of fluorescein inside a microfluidic device. The resulting shape had a 6.38% RSD intensity across the flat profile when imaged with a CCD camera. This flat illumination profile was then used in a two bead multiplex immunoassay where the colour combination and/or the intensities can be used to determine the presence of an antigen. Detection limits of TNF-alpha and IL-6 were comparable with standard ELISA immunoassays.