El presente trabajo resume los resultados del ejercicio realizado para estudiar la variabilidad interlaboratorio del ensayo con estadios larvarios del erizo de mar Paracentrotus lividus. Este ...ejercicio, que se desarrolló en dos fases distintas, incluyó cuatro laboratorios cada uno de los cuales estudió la toxicidad de las seis muestras de sedimento distribuidas. Las muestras, provenientes de distintos puertos de la costa española, se caracterizaron mediante la exposición de embriones del erizo de mar Paracentrotus lividus durante 48 h a los lixiviados de los sedimentos. La Fase I se utilizó para rediseñar las condiciones del ensayo y evitar posibles factores de confusión al interpretar los resultados. Los resultados de la Fase II fueron más homogéneos al clasificar las muestras según la toxicidad registrada, a pesar de la variabilidad en los protocolos de obtención y ensayo de los lixiviados. De acuerdo con estos resultados, el ensayo es adecuado para la caracterización de este tipo de muestras con una variabilidad interlaboratorio similar a la encontrada para otros bioensayos en estudios interlaboratorio previos.
To examine the clinical characteristics and outcomes of cancer patients with venous thromboembolism (VTE) in order to identify factors that place these patients at an increased risk for fatal ...pulmonary embolism (PE) or fatal bleeding.
Registro Informatizado de la Enfermedad Trombo Embólica (RIETE) is a prospective registry of consecutive patients with symptomatic, objectively confirmed, acute VTE.
Up to January 2006, a total of 14 391 patients with symptomatic acute VTE were enrolled in RIETE, of whom 2945 (20%) had cancer. During the 3-month follow-up period the frequency of fatal PE in cancer patients was 2.6%, and that of fatal bleeding 1.0%. These frequencies were significantly higher than in VTE patients without cancer (1.4% and 0.3%, respectively). In patients with cancer, abnormal renal function, metastatic disease, recent major bleeding and recent immobility for >or= 4 days (42% of the 108 patients who died from PE or bleeding had recent immobility) were factors independently associated with an increased risk for both fatal PE and fatal bleeding. In addition, PE diagnosis on admission was an independent risk factor for fatal PE, while body weight < 60 kg was an independent risk factor for fatal bleeding.
Both fatal PE and fatal bleeding are more common in cancer patients with VTE than in those patients without cancer. In cancer patients, abnormal renal function, metastatic disease, recent major bleeding and recent immobility for >or= 4 days are associated with an increased risk for both fatal PE and fatal bleeding.
Abstract
Study question
Does the type of culture medium and its supplementation with protein affect the sensitivity of the Mouse Embryo Assay (MEA) to detect Triton X-100 (TX-100)?
Summary answer
HTF ...medium showed a higher sensitivity than KSOM’s complex formulation, whilst the concentration of protein tested did not have a significant impact on MEA results.
What is known already
TX-100 is a detergent commonly used in the cleaning of production material. Occasionally, small TX-100 traces can remain in the final products, having sublethal effects on embryo development, potentially affecting their viability and implantation potential. The effectiveness of the MEA in detecting low-toxicity levels is not well described due to the lack of methodological standardization or regulation on the appropriateness of different testing protocols.
It is unknown whether media formulation modifies the sensitivity of the MEA. Similarly, media supplementation with albumin is suspected to chelate toxins and decrease the sensitivity of the assay, but no clear relation has been described.
Study design, size, duration
KSOM and HTF were selected and compared as media with complex vs. simple-formulation, respectively. The effect of protein concentration was assessed with both media at 0.5 and 5mg/ml HSA. Both variables were combined to culture embryos in the presence of sublethal concentrations of TX-100, previously established at 0.0005% and 0.0010%. Sensitivity was established by comparing blastocyst formation rates and total cell counts, which were statistically compared between groups to determine the conditions of greatest sensitivity.
Participants/materials, setting, methods
Experiments were performed with freshly-collected mouse zygotes (n = 783). Each TX-100 concentration (0% control, 0.0005%, 0.0010%) was tested with all the proposed medium-protein combinations: KSOM 0.5mg/ml HSA, KSOM 5mg/ml HSA, HTF 0.5 HSA and HTF 5mg/ml HSA. All tests were performed in triplicate in a big-box humidified trigas incubator. Blastocyst formation was assessed after 96h of culture, and the obtained blastocysts were fixed and stained for cell counting.
Main results and the role of chance
No difference was observed in the blastocyst formation rate, nor the total cell counts between any of the control groups, showing that both studied media and protein concentrations were able to successfully support embryo development in vitro.
Based on the expanded blastocyst formation rates, HTF conferred greater sensitivity than KSOM with 5mg/ml HSA at 0.0010% TX-100 (p = 0.0024), and a similar but non-significant trend was observed with 0.5mg/ml (p = 0.0525). Interestingly, increasing HSA to 5mg/ml in HTF reported a higher sensitivity than 0.5mg/ml (p = 0.0399), which was not observed in KSOM. No medium-protein combination was able to detect TX-100 at 0.0005%.
Regarding the cell counting results, each test group cultured with 0.0005% or 0.0010% TX-100 was compared to its own medium-protein combination control. A statistically lower cell number (p < 0.05) was obtained with 0.0010% TX-100 compared to the control groups, regardless of the culture medium or protein concentration used. By contrast, no statistical differences were detected in any of the groups when reducing TX-100 to 0.0005%. Interestingly, with both media and all TX-100 concentrations, the mean cell number per blastocyst was higher when increasing the concentration of HSA from 0.5 to 5mg/ml, whilst this had no effect on the result of the assays.
Limitations, reasons for caution
HTF proved more sensitive than KSOM with 5mg/ml HSA. The limited number of embryos/group could not confirm the statistical significance of this difference with 0.5mg/ml, but a clear trend was observed (p = 0.0525), which might be confirmed by additional replicas. Moreover, the results applicability to other toxicity sources requires further study.
Wider implications of the findings
Numerous culture variables can influence MEA’s sensitivity. Our results suggest that the type of culture medium can have a direct impact on the ability of the MEA to detect certain types of toxins. This information could help detect sublethal amounts of embryotoxic substances, avoiding potential negative clinical events.
Trial registration number
Not applicable
There is evidence for a genetic contribution to chronic periodontitis. In this study, we conducted a genome wide association study among 866 participants of the University of Pittsburgh Dental ...Registry and DNA Repository, whose periodontal diagnosis ranged from healthy (N = 767) to severe chronic periodontitis (N = 99).
Genotypingi of over half-million single nucleotide polymorphisms was determined. Analyses were done twice, first in the complete dataset of all ethnicities, and second including only samples defined as self-reported Whites. From the top 100 results, twenty single nucleotide polymorphisms had consistent results in both analyses (borderline p-values ranging from 1E-05 to 1E-6) and were selected to be tested in two independent datasets derived from 1,460 individuals from Porto Alegre, and 359 from Rio de Janeiro, Brazil. Meta-analyses of the Single nucleotide polymorphisms showing a trend for association in the independent dataset were performed.
The rs1477403 marker located on 16q22.3 showed suggestive association in the discovery phase and in the Porto Alegre dataset (p = 0.05). The meta-analysis suggested the less common allele decreases the risk of chronic periodontitis.
Our data offer a clear hypothesis to be independently tested regarding the contribution of the 16q22.3 locus to chronic periodontitis.
Caso Clínico: Mujer de 33 años de edad con masa pigmentada, asintomática en ángulo iridocorneal de ojo derecho originada en cuerpo ciliar. Diagnóstico de sospecha de melanocitoma de cuerpo ciliar se ...decidió observación. Después de 36 meses de seguimiento presenta dolor, reacción inflamatoria y aumento incontrolable de presión intraocular que se diagnostica de glaucoma melanocitomalítico. Se realiza escisión del tumor mediante iridociclectomía «ab-externo» y el estudio histopatológico demostró melanocitoma necrótico. Discusión: El melanocitoma de cuerpo ciliar es un tumor pigmentado benigno, poco frecuente que puede presentar extensión a cámara anterior. El diagnóstico diferencial se realiza fundamentalmente con melanoma de cuerpo ciliar, que implica un diferente tratamiento y pronóstico.
A full comprehension of colorimetric relationships within and between teeth is key for aesthetic success of a dental restoration. In this sense, hyperspectral imaging can provide point-wise reliable ...measurements of the tooth surface, which can serve for this purpose. The aim of this study was to use a hyperspectral imaging system for the colorimetric characterization of 4 in-vivo maxillary anterior teeth and to cross-check the results with similar studies carried out with other measuring systems in order to validate the proposed capturing protocol. Hyperspectral reflectance images (Specim IQ), of the upper central (UCI) and lateral incisors (ULI), were captured on 30 participants. CIE-L*a*b* values were calculated for the incisal (I), middle (M) and cervical (C) third of each target tooth. ΔEab* and ΔE00 total color differences were computed between different tooth areas and adjacent teeth, and evaluated according to the perceptibility (PT) and acceptability (AT) thresholds for dentistry. Non-perceptible color differences were found between UCIs and ULIs. Mean color differences between UCI and ULI exceeded AT (ΔEab* = 7.39–7.42; ΔE00 = 5.71–5.74) in all cases. Large chromatic variations between I, M and C areas of the same tooth were registered (ΔEab* = 5.01–6.07 and ΔE00 = 4.07–5.03; ΔEab* = 5.80–8.16 and ΔE00 = 4.37–5.15; and ΔEab* = 5.42–5.92 and ΔE00 = 3.87–4.16 between C and M, C and I and M and I, respectively). The use of a hyperspectral camera has proven to be a reliable and effective method for color evaluation of in-vivo natural teeth.
Growth hormone-releasing hormone (GHRH) has a crucial role in growth hormone (GH) secretion, but little is known about its production by adipocytes and its involvement in adipocyte metabolism.
To ...determine whether GHRH and its receptor (GHRH-R) are present in human adipocytes and to study their levels in obesity. Also, to analyze the effects of GHRH on human adipocyte differentiation and lipolysis.
GHRH/GHRH-R and GH/GH-R mRNA expression levels were analyzed in human mature adipocytes from non-obese and morbidly obese subjects. Human mesenchymal stem cells (HMSC) were differentiated to adipocytes with GHRH (10
-10
M). Adipocyte differentiation, lipolysis and gene expression were measured and the effect of GH-R silencing was determined.
Mature adipocytes from morbidly obese subjects showed a higher expression of GHRH and GH-R, and a lower expression of GHRH-R and GH than non-obese subjects (P<0.05). A total of 10
-10
M GHRH induced an inhibition of lipid accumulation and PPAR-γ expression (P<0.05), and an increase in glycerol release and HSL expression (P<0.05) in human differentiated adipocytes. A total of 10
-10
M GHRH decreased GHRH-R expression in human differentiated adipocytes (P<0.05). A total of 10
-10
M GHRH increased GH and GH-R expression in human differentiated adipocytes (P<0.05). The effects of GHRH at 10
M on adipocyte differentiation and lipolysis were blocked when GH-R expression was silenced.
GHRH and GHRH-R are expressed in human adipocytes and are negatively associated. GHRH at low doses may exert an anti-obesity effect by inhibiting HMSC differentiation in adipocytes and by increasing adipocyte lipolysis in an autocrine or paracrine pathway. These effects are mediated by GH and GH-R.
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