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•Passive sampling allowed for largely matrix-free transfer of pollutants from biota tissues.•Non-destructive cleanup of extracts still changed the composition of the extract.•Cleanup ...improved the bioanalytical measurement thanks to lower reduction of bioavailability.•Effects were highest in extracts from liver > kidney and brain > blubber.
We used in-tissue passive equilibrium sampling using the silicone polydimethylsiloxane (PDMS) to transfer chemical mixtures present in organs from marine mammals with lipid contents between 2.3 and 99%into in vitro bioassays. Tissues from five harbor porpoises (Phocoena phocoena), one harbor seal (Phoca vitulina) and one orca (Orcinus orca) from the North and Baltic Seas were sampled until thermodynamic equilibrium was reached. Mixture effects were quantified with cellular reporter gene bioassays targeting the activation of the aryl hydrocarbon receptor (AhR-CALUX), the peroxisome proliferator-activated receptor gamma (PPARγ-bla) and the oxidative stress response (AREc32), with parallel cytotoxicity measurements in all assays. After removing co-extracted lipids and other matrix residues with a non-destructive cleanup method (freeze-out of acetonitrile extract followed by a primary secondary amine sorbent extraction), the activation of the PPARγ and AREc32 were reduced by factors of on average 4.3 ± 0.15 (n = 22) and 2.5 ± 0.23 (n = 18), respectively, whereas the activation of the AhR remained largely unaltered: 1.1 ± 0.075 (n = 6). The liver extracts showed the highest activation, followed by the corresponding kidney and brain extracts, and the blubber extracts of the animals were the least active ones. The activation of the PPARγ by the liver extracts was reduced after cleanup by a factor of 11 ± 0.26 (n = 7) and the AREc32 activity by a factor of 1.9 ± 0.32 (n = 4). The blubber extracts did not activate the AhR up to concentrations where cytotoxicity occurred or up to an acceptable lipid volume fraction of 0.27% as derived from earlier work, whereas all liver extracts that had undergone cleanup activated the AhR. The developed in-tissue passive sampling approach allows a direct comparison of the bioassay responses between different tissues without further normalization and serves as a quantitative method suitable for biomonitoring of environmental biota samples.
The present study complements work on mixture effects measured with
bioassays of passive equilibrium sampling extracts using the silicone polydimethylsiloxane (PDMS) in organs from marine mammals ...with chemical profiling. Blubber, liver, kidney and brain tissues of harbor porpoise (
), harbor seal (
), ringed seal (
) and orca (
) from the North and Baltic Seas were investigated. We analyzed 117 chemicals including legacy and emerging contaminants using gas chromatography-high resolution mass spectrometry and quantified 70 of those chemicals in at least one sample. No systematic differences between the organs were found. Only for single compounds a clear distribution pattern was observed. For example, 4,4'-dichlorodiphenyltrichloroethane, enzacamene and etofenprox were mainly detected in blubber, whereas tonalide and the hexachlorocyclohexanes were more often found in liver. Furthermore, we compared the chemical profiling with the bioanalytical results using an iceberg mixture model, evaluating how much of the biological effect could be explained by the analyzed chemicals. The mixture effect predicted from the quantified chemical concentrations explained 0.014-83% of the aryl hydrocarbon receptor activating effect (AhR-CALUX), but less than 0.13% for the activation of the oxidative stress response (AREc32) and peroxisome-proliferator activated receptor (PPARγ). The quantified chemicals also explained between 0.044-45% of the cytotoxic effect measured with the AhR-CALUX. The largest fraction of the observed effect was explained for the orca, which was the individuum with the highest chemical burden. This study underlines that chemical analysis and bioassays are complementary to comprehensively characterize the mixture exposome of marine mammals.
Extraction of chemicals from biota leads to co-extraction of lipids. When dosing such extracts into in vitro bioassays, co-dosed lipids act as an additional phase that can reduce the bioavailability ...of the chemicals and the apparent sensitivity of the assay. Equilibrium partitioning between medium, cells, and co-dosed lipids was described with an existing equilibrium partitioning model for cell-based bioassays extended by an additional lipid phase. We experimentally investigated the influence of co-dosed lipids on the effects elicited by four test chemicals of different hydrophobicity in two bioassays, indicative of the aryl hydrocarbon receptor and oxidative stress response (AREc32). The partitioning model explained the effect of the test chemicals in the presence of spiked triolein within a factor of 0.33–5.83 between the measured and predicted effect concentration (EC). We applied the model to marine mammal blubber extracted with silicone. Extracts dosed in the AREc32 bioassay showed a linear increase of apparent EC with increasing lipid fraction. The partitioning model was used to interpret the role of the co-extracted lipid. A quantitative lipid correction of bioassay results in the presence of co-dosed lipids was possible for known compounds and defined mixtures, while we could only estimate a range for mixtures of unknown chemicals.
Detailed post-mortem investigations including the auditory pathway are needed to advance our understanding of how underwater noise and other stressors affect hearing in cetaceans. A 12-year-old ...female porpoise (
Phocoena phocoena
) stranded alive in June 2021 at the German Baltic Sea coast and died some hours later. The most significant pathological findings were lesions caused by a severe aspergillosis that spread from the lung and pulmonary lymph node to the cerebellum. Based on molecular sequencing, the fungus was identified as
Aspergillus fumigatus.
Severe pyogranulomatous and necrotizing inflammation was diagnosed in the lung and the associated lymph node. In the left part of the cerebellum, focal, severe purulent and necrotizing meningoencephalitis with intralesional fungal structures was confirmed histologically. In addition, multifocal, severe, chronic, granulomatous, and eosinophilic gastritis with intralesional parasite structures was found in the stomach. Parallel stripes (linear skin markings) were detected along the caudal part of both body sides, which have not been previously described for harbor porpoises. Inner ear analysis revealed evidence of focal loss of outer hair cells in several regions from 120 to 580 µm from the apex of the right cochlea using immunofluorescence. The evidence of low-frequency hearing impairment was compatible with noise-induced hearing loss. This is the first case of concurrent presumptive noise-induced hearing loss and unrelated aspergillosis in a free-ranging harbor porpoise.
The present study complements work on mixture effects measured with
in vitro
bioassays of passive equilibrium sampling extracts using the silicone polydimethylsiloxane (PDMS) in organs from marine ...mammals with chemical profiling. Blubber, liver, kidney and brain tissues of harbor porpoise (
Phocoena phocoena
), harbor seal (
Phoca vitulina
), ringed seal (
Phoca hispida
) and orca (
Orcinus orca
) from the North and Baltic Seas were investigated. We analyzed 117 chemicals including legacy and emerging contaminants using gas chromatography-high resolution mass spectrometry and quantified 70 of those chemicals in at least one sample. No systematic differences between the organs were found. Only for single compounds a clear distribution pattern was observed. For example, 4,4′-dichlorodiphenyltrichloroethane, enzacamene and etofenprox were mainly detected in blubber, whereas tonalide and the hexachlorocyclohexanes were more often found in liver. Furthermore, we compared the chemical profiling with the bioanalytical results using an iceberg mixture model, evaluating how much of the biological effect could be explained by the analyzed chemicals. The mixture effect predicted from the quantified chemical concentrations explained 0.014-83% of the aryl hydrocarbon receptor activating effect (AhR-CALUX), but less than 0.13% for the activation of the oxidative stress response (AREc32) and peroxisome-proliferator activated receptor (PPARγ). The quantified chemicals also explained between 0.044-45% of the cytotoxic effect measured with the AhR-CALUX. The largest fraction of the observed effect was explained for the orca, which was the individuum with the highest chemical burden. This study underlines that chemical analysis and bioassays are complementary to comprehensively characterize the mixture exposome of marine mammals.
The analysis of mixtures of environmental contaminants from marine mammal organs revealed distinct distribution patterns for single compounds. The combination of chemical analysis and bioassays can comprehensively characterize the mixture exposome.
The aims of this study were to develop a comprehensive cardiovascular magnetic resonance (CMR) approach to diastolic dysfunction (DD) grading and to evaluate the accuracy of CMR in the diagnosis of ...DD compared with echocardiography.
Left ventricular DD is routinely assessed using echocardiography.
Consecutive clinically referred patients (n = 46; median age 59 years; interquartile range: 46 to 68 years; 33% women) underwent both conventional echocardiography and CMR. CMR diastolic transmitral velocities (E and A) and myocardial tissue velocity (e′) were measured during breath-hold using a validated high–temporal resolution radial sector-wise golden-angle velocity-encoded sequence. CMR pulmonary artery pressure was estimated from 4-dimensional flow analysis of blood flow vortex duration in the pulmonary artery. CMR left atrial volume was measured using the biplane long-axis area-length method. Both CMR and echocardiographic data were used to perform blinded grading of DD according to the 2016 joint American and European recommendations.
Grading of DD by CMR agreed with that by echocardiography in 43 of 46 cases (93%), of which 9% were normal, 2% indeterminate, 63% grade 1 DD, 4% grade 2 DD, and 15% grade 3 DD. There was a very good categorical agreement, with a weighted Cohen kappa coefficient of 0.857 (95% confidence interval: 0.73 to 1.00; p < 0.001).
A comprehensive CMR protocol for grading DD encompassing diastolic blood and myocardial velocities, estimated pulmonary artery pressure, and left atrial volume showed very good agreement with echocardiography.
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Pulmonary hypertension is definitively diagnosed by the measurement of mean pulmonary artery (PA) pressure (mPAP) using right heart catheterization. Cardiovascular magnetic resonance (CMR) ...four-dimensional (4D) flow analysis can estimate mPAP from blood flow vortex duration in the PA, with excellent results. Moreover, the peak systolic tricuspid regurgitation (TR) pressure gradient (TRPG) measured by Doppler echocardiography is commonly used in clinical routine to estimate systolic PA pressure. This study aimed to compare CMR and echocardiography with regards to quantitative and categorical agreement, and diagnostic yield for detecting increased PA pressure.
Consecutive clinically referred patients (n = 60, median interquartile range age 60 48-68 years, 33% female) underwent echocardiography and CMR at 1.5 T (n = 43) or 3 T (n = 17). PA vortex duration was used to estimate mPAP using a commercially available time-resolved multiple 2D slice phase contrast three-directional velocity encoded sequence covering the main PA. Transthoracic Doppler echocardiography was performed to measure TR and derive TRPG. Diagnostic yield was defined as the fraction of cases in which CMR or echocardiography detected an increased PA pressure, defined as vortex duration ≥15% of the cardiac cycle (mPAP ≥25 mmHg) or TR velocity > 2.8 m/s (TRPG > 31 mmHg).
Both CMR and echocardiography showed normal PA pressure in 39/60 (65%) patients and increased PA pressure in 9/60 (15%) patients, overall agreement in 48/60 (80%) patients, kappa 0.49 (95% confidence interval 0.27-0.71). CMR had a higher diagnostic yield for detecting increased PA pressure compared to echocardiography (21/60 (35%) vs 9/60 (15%), p < 0.001). In cases with both an observable PA vortex and measurable TR velocity (34/60, 56%), TRPG was correlated with mPAP (R
= 0.65, p < 0.001).
There is good quantitative and fair categorical agreement between estimated mPAP from CMR and TRPG from echocardiography. CMR has higher diagnostic yield for detecting increased PA pressure compared to echocardiography, potentially due to a lower sensitivity of echocardiography in detecting increased PA pressure compared to CMR, related to limitations in the ability to adequately visualize and measure the TR jet by echocardiography. Future comparison between echocardiography, CMR and invasive measurements are justified to definitively confirm these findings.
Myeloid neoplasms and acute leukemias derive from the clonal expansion of hematopoietic cells driven by somatic gene mutations. Although assessment of morphology plays a crucial role in the ...diagnostic evaluation of patients with these malignancies, genomic characterization has become increasingly important for accurate diagnosis, risk assessment, and therapeutic decision making. Conventional cytogenetics, a comprehensive and unbiased method for assessing chromosomal abnormalities, has been the mainstay of genomic testing over the past several decades and remains relevant today. However, more recent advances in sequencing technology have increased our ability to detect somatic mutations through the use of targeted gene panels, whole-exome sequencing, whole-genome sequencing, and whole-transcriptome sequencing or RNA sequencing. In patients with myeloid neoplasms, whole-genome sequencing represents a potential replacement for both conventional cytogenetic and sequencing approaches, providing rapid and accurate comprehensive genomic profiling. DNA sequencing methods are used not only for detecting somatically acquired gene mutations but also for identifying germline gene mutations associated with inherited predisposition to hematologic neoplasms. The 2022 International Consensus Classification of myeloid neoplasms and acute leukemias makes extensive use of genomic data. The aim of this report is to help physicians and laboratorians implement genomic testing for diagnosis, risk stratification, and clinical decision making and illustrates the potential of genomic profiling for enabling personalized medicine in patients with hematologic neoplasms.
Complementing the recently published Blood articles outlining the 2022 International Consensus Classifications for hematological malignancies (Vol. 140, Issue 11), this pair of Special Reports illustrates how molecular pathology can be applied to precision medicine. de Leval and colleagues summarize the potential of DNA sequencing of tumors and cell-free plasma, epigenetic profiling, and single-cell analyses to inform clinical decision-making about diagnosis, prognosis, and treatment for patients with lymphoid neoplasms. Similarly, Duncavage and colleagues cover genomic profiling for myeloid neoplasms and the acute leukemias, focusing principally on somatic changes but also with emphasis on the emerging importance of germline gene mutations in certain diseases. Both articles provide up-to-date references for how to apply genomic information to practice.
Abstract
Background
the European Union of Medical Specialists (UEMS-GMS) recommendations for training in Geriatric Medicine were published in 1993. The practice of Geriatric Medicine has developed ...considerably since then and it has therefore become necessary to update these recommendations.
Methods
under the auspices of the UEMS-GMS, the European Geriatric Medicine Society (EuGMS) and the European Academy of Medicine of Ageing (EAMA), a group of experts, representing all member states of the respective bodies developed a new framework for education and training of specialists in Geriatric Medicine using a modified Delphi technique. Thirty-two expert panel members from 30 different countries participated in the process comprising three Delphi rounds for consensus. The process was led by five facilitators.
Results
the final recommendations include four different domains: ‘General Considerations’ on the structure and aim of the syllabus as well as quality indicators for training (6 sub-items), ‘Knowledge in patient care’ (36 sub-items), ‘Additional Skills and Attitude required for a Geriatrician’ (9 sub-items) and a domain on ‘Assessment of postgraduate education: which items are important for the transnational comparison process’ (1 item).
Conclusion
the current publication describes the development of the new recommendations endorsed by UEMS-GMS, EuGMS and EAMA as minimum training requirements to become a geriatrician at specialist level in EU member states.
The analysis of rare chromosomal translocations in myeloproliferative disorders has highlighted the importance of aberrant tyrosine kinase signaling in the pathogenesis of these diseases. Here we ...have investigated samples from 679 patients and controls for the nonreceptor tyrosine kinase JAK2 V617F mutation. Of the 480 myeloproliferative disorder (MPD) samples, the proportion of positive cases per disease subtype was 30 (20%) of 152 for atypical or unclassified MPD, 2 of 134 (2%) for idiopathic hypereosinophilic syndrome, 58 of 72 (81%) for polycythemia vera, 24 of 59 (41%) essential thrombocythemia (ET), and 15 of 35 (43%) for idiopathic myelofibrosis. V617F was not identified in patients with systemic mastocytosis (n = 28), chronic or acute myeloid leukemia (n = 35), secondary erythrocytosis (n = 4), or healthy controls (n = 160). Homozygosity for V617F was seen in 43% of mutant samples and was closely correlated with chromosome 9p uniparental disomy. Homozygosity was significantly less common in ET compared with other MPD subtypes. In 53 cases analyzed, the median level of PRV1 expression was significantly higher in V617F-positive cases compared with cases without the mutation. We conclude that V617F is widespread in MPDs. Detection of this acquired mutation is likely to have a major impact on the way patients with MPD are diagnosed, as well as serving as an obvious target for signal transduction therapy. (Blood. 2005;106:2162-2168)
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