Physicochemical Characterization of the Oral Biotherapeutic Drug IMUNOR®.

IMUNOR is an oral biotherapeutic drug that had been developed, registered, and approved in 1997 in the Czech Republic and Slovakia. IMUNOR is a dialyzable leukocyte extract (DLE) prepared from swine leukocytes. It is characterized as a mixture of small peptides with molecular weights smaller than 12 kDa and a specific portion of nucleotides. The medical uses of IMUNOR include therapeutic applications within its registered range of indications, primarily for the treatment of immunodeficiencies, allergies, and certain acute or relapsing bacterial infections in adults and children. Despite the long-term clinical application of DLE, with strong evidence of positive therapeutic effects and no serious side effects, a detailed physicochemical specification of this mixture was lacking. We developed several methods for more in-depth physicochemical characterization of IMUNOR, including a spectrophotometric method for quantification of the total protein concentration and total DNA concentration in a mixture, several chromatographic methods for identification of individual components present in significant concentrations in IMUNOR, such as HPLC methods and the Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis method, and characterization of amino acid composition of this mixture. For the investigation of the variability among different batches of IMUNOR, five to nine representative batches from a standard manufacturing process on an industrial scale were utilized. Using the analytical methods, we verified and confirmed the batch-to-batch reproducibility of the biological product IMUNOR.

Cross sectional study on exposure to BPA and its analogues and semen parameters in Czech men.

Exposure to bisphenols has been found to have adverse effects on male reproductive function in animals. Human exposure to bisphenols is widespread. Bisphenol A (BPA) and its analogues, including bisphenol S (BPS), bisphenol F (BPF), and bisphenol AF (BPAF) are utilized in various consumer products such as food contact materials and dental resins. The effects of these compounds on male fertility and spermatogenesis are unclear and findings from human studies are inconsistent. In this cross-sectional study, we evaluated the influence of BPA, BPS, BPF, BPAF (BPs) measured in semen on number of spermatozoa, total motility, progressive motility, morphology, and DNA fragmentation. We also examined the association of bisphenols (BPs) exposure with patients' occupation. A total of 358 patients aged 17-62 years with BMI 18-42 were included in the study from 2019 to 2021. BPs were extracted using solvent extraction followed by preconcentration step and determined by high-performance liquid chromatography and tandem mass spectrometry (LC/MSMS). Bisphenols were detected in 343 from 349 analysed samples (98.3% of all the samples). In 6 samples, the concentration of all BPs was under the limit of detection and in 20 samples under the limit of quantification. We did not find a statistically significant relationship between occupation and BPs. However, we observed significant correlations between the concentration of BPA and a lower motility and normal morphology. For BPS, a significant correlation with a lower ejaculate volume and a lower total sperm count was found. BPF and BPAF were detected only in 14.3% and 23.9% of samples, respectively. For BPF and BPAF, no significant correlations with spermiogram parameters were observed. Our results show that BPs are widespread in the male population (more than 90% of analysed samples), independently of an occupation and in case of BPA and BPS having a negative impact on spermiogram parameters.

Current-use pesticide exposure pathways in Czech adults and children from the CELSPAC-SPECIMEn cohort.

INTRODUCTION: In this study, we aimed to characterise exposure to pyrethroids, organophosphates, and tebuconazole through multiple pathways in 110 parent-child pairs participating in the CELSPAC-SPECIMEn study. METHODS: First, we estimated the daily intake (EDI) of pesticides based on measured urinary metabolites. Second, we compared EDI with estimated pesticide intake from food. We used multiple linear regression to identify the main predictors of urinary pesticide concentrations. We also assessed the relationship between urinary pesticide concentrations and organic and non-organic food consumption while controlling for a range of factors. Finally, we employed a model to estimate inhalation and dermal exposure due to spray drift and volatilization after assuming pesticide application in crop fields. RESULTS: EDI was often higher in children in comparison to adults, especially in the winter season. A comparison of food intake estimates and EDI suggested diet as a critical pathway of tebuconazole exposure, less so in the case of organophosphates. Regression models showed that consumption per g of peaches/apricots was associated with an increase of 0.37% CI [0.23% to 0.51%] in urinary tebuconazole metabolite concentrations. Consumption of white bread was associated with an increase of 0.21% CI [0.08% to 0.35%], and consumption of organic strawberries was inversely associated (-61.52% CI [-79.34% to -28.32%]), with urinary pyrethroid metabolite concentrations. Inhalation and dermal exposure seemed to represent a relatively small contribution to pesticide exposure as compared to dietary intake. CONCLUSION: In our study population, findings indicate diet plays a significant role in exposure to the analysed pesticides. We found an influence of potential exposure due to spray drift and volatilization among the subpopulation residing near presumably sprayed crop fields to be minimal in comparison. However, the lack of data indicating actual spraying occurred during the critical 24-hour period prior to urine sample collection could be a significant contributing factor.

Has Regulatory Action Reduced Human Exposure to Flame Retardants?

Flame retardant (FR) exposure has been linked to several environmental and human health effects. Because of this, the production and use of several FRs are regulated globally. We reviewed the available records of polybrominated diphenyl ethers (PBDEs) and hexabromocyclododecanes (HBCDDs) in human breast milk from literature to evaluate the efficacy of regulation to reduce the exposure of FRs to humans. Two-hundred and seven studies were used for analyses to determine the spatial and temporal trends of FR exposure. North America consistently had the highest concentrations of PBDEs, while Asia and Oceania dominated HBCDD exposure. BDE-49 and -99 indicated decreasing temporal trends in most regions. BDE-153, with a longer half-life than the aforementioned isomers, typically exhibited a plateau in breast milk levels. No conclusive trend could be established for HBCDD, and insufficient information was available to determine a temporal trend for BDE-209. Breakpoint analyses indicated a significant decrease in BDE-47 and -99 in Europe around the time that regulation has been implemented, suggesting a positive effect of regulation on FR exposure. However, very few studies have been conducted globally (specifically in North America) after 2013, during the time when the most recent regulations have been implemented. This meta-analysis provides insight into global trends in human exposure to PBDEs and HBCDD, but the remaining uncertainty highlights the need for ongoing evaluation and monitoring, even after a compound group is regulated.

Spatial and Temporal Trends of Persistent Organic Pollutants across Europe after 15 Years of MONET Passive Air Sampling.

The Global Monitoring Plan of the Stockholm Convention on Persistent Organic Pollutants (POPs) was established to generate long-term data necessary for evaluating the effectiveness of regulatory measures at a global scale. After 15 years of passive air monitoring (2003-2019), MONET is the first network to produce sufficient data for the analysis of continuous long-term temporal trends of POPs in air across the entire European continent. This study reports long-term concentrations of 20 POPs monitored at 32 sites in 27 European countries. As of January 1, 2019, the concentration ranges (pg/m3) were 1.1-52.8 (∑6PCB), 0.3-8.5 (∑12dl-PCB), 0.007-0.175 (∑17PCDD/F), 0.02-2.2 (∑9PBDE), 0.4-24.7 (BDE 209), 0.5-247 (∑6DDT), 1.7-818 (∑4HCH), 15.8-74.7 (HCB), and 5.9-21.5 (PeCB). Temporal trends indicate that concentrations of most POPs have declined significantly over the past 15 years, with median annual decreases ranging from -8.0 to -11.5% (halving times of 6-8 years) for ∑6PCB, ∑17PCDD/F, HCB, PeCB, and ∑9PBDE. Furthermore, no statistically significant differences were observed in either the trends or the concentrations of specific POPs at sites in Western Europe (WEOG) compared to sites in Central and Eastern Europe (CEE), which suggests relatively uniform compound-specific distribution and removal at the continental scale.

Impact of PFAS exposure on prevalence of immune-mediated diseases in adults in the Czech Republic.

BACKGROUND: Per- and polyfluoroalkyl substances (PFASs) are emerging environmental contaminants with multiple hazardous properties including immunomodulation potency. Human exposure to PFASs has been associated with various immune-mediated diseases and outcomes. This study aimed to investigate the association between PFAS exposure and immune-mediated diseases such as allergies, eczemas, and autoimmune diseases in a population of adults in the Czech Republic. METHODS: This study included 309 adults from the Central European Longitudinal Study of Parents and Children: Young Adults (CELSPAC: YA). 12 PFASs were measured in participants' serum by HPLC-MS/MS, 3 PFASs were removed from the subsequent analyses due to low detection frequency. The associations of 9 PFASs with 9 immune-mediated diseases were assessed by logistic regression. Furthermore, Bayesian kernel machine regression (BKMR) was used to estimate the effect of the PFAS mixture on immune-mediated diseases. All analyses were adjusted for sex, age, BMI, smoking, education, and family history of immune-mediated diseases. In cases of a statistically significant interaction of PFASs and sex, stratified analyses were performed for men and women. RESULTS: Perfluorooctanoate (PFOA) and perfluorooctane sulfonate (PFOS) were negatively associated with both atopic eczema (OR per IQR increase 0.58 (95% CI 0.37-0.90) for PFOA and 0.56 (0.32-0.95) for PFOS) and contact dermatitis (0.37 (0.16-0.85) for PFOA and 0.33 (0.11-0.94) for PFOS). Perfluoroundecanoate (PFUnDA) was negatively associated with pollen, dust, and mite allergy (0.62 (0.43-0.89)). BKMR modelling showed a negative tendency in the overall effect of PFAS mixture on immune-health outcomes. Based on the stratified analysis, sex was suggested to be an effect modifier in the association of PFOS and atopic eczema. CONCLUSION: Our results contribute to the body of literature that observes the immunosuppressive effect of PFAS exposure during eczemas and allergies, both for PFASs individually and as a mixture.

Time-trends in human urinary concentrations of phthalates and substitutes DEHT and DINCH in Asian and North American countries (2009-2019).

BACKGROUND: Many phthalates are environmental pollutants and toxic to humans. Following phthalate regulations, human exposure to phthalates has globally decreased with time in European countries, the US and Korea. Conversely, exposure to their substitutes DEHT and/or DINCH has increased. In other countries, including China, little is known on the time-trends in human exposure to these plasticizers. OBJECTIVE: We aimed to estimate time-trends in the urinary concentrations of phthalates, DEHT, and DINCH metabolites, in general population from non-European countries, in the last decade. METHODS: We compiled human biomonitoring (HBM) data from 123 studies worldwide in a database termed "PhthaLit". We analyzed time-trends in the urinary concentrations of the excreted metabolites of various phthalates as well as DEHT and DINCH per metabolite, age group, and country/region, in 2009-2019. Additionally, we compared urinary metabolites levels between continents. RESULTS: We found solid time-trends in adults and/or children from the US, Canada, China and Taiwan. DEHP metabolites decreased in the US and Canada. Conversely in Asia, 5oxo- and 5OH-MEHP (DEHP metabolites) increased in Chinese children. For low-weight phthalates, the trends showed a mixed picture between metabolites and countries. Notably, MnBP (a DnBP metabolite) increased in China. The phthalate substitutes DEHT and DINCH markedly increased in the US. SIGNIFICANCE: We addressed the major question of time-trends in human exposure to phthalates and their substitutes and compared the results in different countries worldwide. IMPACT: Phthalates account for more than 50% of the plasticizer world market. Because of their toxicity, some phthalates have been regulated. In turn, the consumption of non-phthalate substitutes, such as DEHT and DINCH, is growing. Currently, phthalates and their substitutes show high detection percentages in human urine. Concerning time-trends, several studies, mainly in Europe, show a global decrease in phthalate exposure, and an increase in the exposure to phthalate substitutes in the last decade. In this study, we address the important question of time-trends in human exposure to phthalates and their substitutes and compare the results in different countries worldwide.

Exposure to flame retardants in European children - Results from the HBM4EU aligned studies.

Many legacy and emerging flame retardants (FRs) have adverse human and environmental health effects. This study reports legacy and emerging FRs in children from nine European countries from the HBM4EU aligned studies. Studies from Belgium, Czech Republic, Germany, Denmark, France, Greece, Slovenia, Slovakia, and Norway conducted between 2014 and 2021 provided data on FRs in blood and urine from 2136 children. All samples were collected and analyzed in alignment with the HBM4EU protocols. Ten halogenated FRs were quantified in blood, and four organophosphate flame retardants (OPFR) metabolites quantified in urine. Hexabromocyclododecane (HBCDD) and decabromodiphenyl ethane (DBDPE) were infrequently detected (<16% of samples). BDE-47 was quantified in blood from Greece, France, and Norway, with France (0.36 ng/g lipid) having the highest concentrations. BDE-153 and -209 were detected in <40% of samples. Dechlorane Plus (DP) was quantified in blood from four countries, with notably high median concentrations of 16 ng/g lipid in Slovenian children. OPFR metabolites had a higher detection frequency than other halogenated FRs. Diphenyl phosphate (DPHP) was quantified in 99% of samples across 8 countries at levels ∼5 times higher than other OPFR metabolites (highest median in Slovenia of 2.43 ng/g lipid). FR concentrations were associated with lifestyle factors such as cleaning frequency, employment status of the father of the household, and renovation status of the house, among others. The concentrations of BDE-47 in children from this study were similar to or lower than FRs found in adult matrices in previous studies, suggesting lower recent exposure and effectiveness of PBDE restrictions.

City-scale assessment of long-term air quality impacts on the respiratory and cardiovascular health.

Background: The impact of the urban environment on human health is a contemporary subject of environmental research. Air pollution is often considered a leading environmental driver. However, a plethora of other factors within the urban exposome may be involved. At the same time, the resolution of spatial data is also an important facet to consider. Generally, systematic tools for accurate health risk assessment in the urban environment are missing or are not implemented. Methods: The long-term impact of air quality (PM10, PM2.5, NO2, benzene, and SO2) on respiratory and cardiovascular health was assessed with a log-linear model. We used the most accurate health data in high city scale spatial resolution over the period 2010 to 2018. Selected external exposome parameters were also included in the analysis. Results: Statistically significant associations between air pollution and the health of the urban population were found. The strongest association was between benzene and the incidence of bronchitis in the adult population [RR 1.552 95% CI (1.415-1.704) per 0.5 µg/m3 change in benzene concentration]. A similar relation was observed between NO2 and the same health condition [RR 1.483 95% CI (1.227-1.792) per 8.9 µg/m3 of change in NO2]. Other weaker associations were also found between asthma in children and PMs, NO2, or benzene. Cardiovascular-related hospitalizations in the general population were linked with NO2 [RR 1.218 95% CI (1.119-1.325) per 9.7 µg/m3 change in NO2]. The remaining pollutants were slightly less but still significantly associated with cardiovascular-related hospitalizations. Conclusion: Our findings are mostly highly statistically significant (p ≤ 0.001) and are in line with current literature on the adverse effects of air pollution on the human population. The results highlight the need for continual improvements in air quality. We propose the implementation of this approach as a systematic tool for the investigation of possible health risks over a long period of time. However, further research involving other variables is an essential step toward understanding the complex urban exposome and its implications for human health. An increase in data spatial resolution is especially important in this respect as well as for improving city health risk management.

Central Venous Catheter Cannulation in Pediatric Anesthesia and Intensive Care: A Prospective Observational Trial.

Currently, ultrasound-guided central venous catheter (CVC) insertion is recommended in pediatric patients. However, the clinical practice may vary. The primary aim of this study was the overall success rate and the first attempt success rate in ultrasound-guided CVC insertion versus anatomic-based CVC insertion in pediatric patients. The secondary aim was the incidence of associated complications and the procedural time. The physician could freely choose the cannulation method and venous approach. Data were collected for 10 months. Overall, 179 patients were assessed for eligibility and 107 patients were included. In almost half of the patients (48.6%), the percutaneous puncture was performed by real-time ultrasound navigation. In 51.4% of the patients, the puncture was performed by the landmark method. The overall success rate was 100% (n = 52) in the real-time ultrasound navigation group, 96.4% (n = 53) in the landmark insertion group, (p = 0.496). The first percutaneous puncture success rate was 57.7% (n = 30) in the real-time ultrasound navigation group and 45.5% (n = 25) in the landmark insertion group, (p = 0.460). The data show a higher overall success rate and the first success rate in the US-guided CVC insertion group, but the difference was not statistically significant.

The Bisphenols Found in the Ejaculate of Men Does Not Pass through the Testes.

Exposure to bisphenols is related to negative effects on male reproduction. The bisphenols exposure is associated with several modes of action including negative impact on the blood-testis barrier (BTB) in testes or direct effect on spermatozoa. Bisphenols have been detected in human seminal plasma, but the possible mechanism of seminal transfer of bisphenols is not clear. Some authors consider the transfer through the blood-testis barrier to be crucial. Therefore, in this work, we compared normozoospermic men and men after vasectomy who have interrupted vas deferens and their ejaculate does not contain testicular products. We measured the concentration of bisphenol A (BPA), bisphenol S (BPS) and bisphenol F (BPF) in the urine and seminal plasma of these men using liquid chromatography tandem mass spectrometry (LC/MSMS). We found that the ratio of urinary and seminal plasma content of bisphenols did not differ in normozoospermic men or men after vasectomy. From the obtained data, it can be concluded that the pathways of transport of bisphenols into seminal plasma are not primarily through the testicular tissue, but this pathway is applied similarly to other routes of transmission by a corresponding ejaculate volume ratio. To a much greater extent than through testicular tissue, bisphenols enter the seminal plasma mainly as part of the secretions of the accessory glands.

Comparability of semivolatile organic compound concentrations from co-located active and passive air monitoring networks in Europe.

Passive air sampling (PAS) has been used to monitor semivolatile organic compounds (SVOCs) for the past 20 years, but limitations and uncertainties persist in the derivation of effective sampling volumes, sampling rates, and concentrations. As a result, the comparability of atmospheric levels measured by PAS and concentrations measured by active air sampling (AAS) remains unclear. Long-term PAS data, without conversion into concentrations, provide temporal trends that are similar to, and consistent with, trends from AAS data. However, for more comprehensive environmental and human health assessments of SVOCs, it is also essential to harmonize and pool air concentration data from the major AAS and PAS monitoring networks in Europe. To address this need, we calculated and compared concentration data for 28 SVOCs (including organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), and polycyclic aromatic hydrocarbons (PAHs)) at the six monitoring sites in Europe with 10 years of co-located AAS (EMEP) and PAS (MONET) data: Birkenes, Kosetice, Pallas, Råö, Stórhöfði, and Zeppelin. Atmospheric SVOC concentrations were derived from PAS data using the two most common computation models. Long-term agreement between the AAS and PAS data was strong for most SVOCs and sites, with 79% of the median PAS-derived concentrations falling within a factor of 3 of their corresponding AAS concentrations. However, in both models it is necessary to set a sampler-dependent correction factor to prevent underestimation of concentrations for primarily particle-associated SVOCs. In contrast, the models overestimate concentrations at sites with wind speeds that consistently exceed 4 m s-1. We present two recommendations that, if followed, allow MONET PAS to provide sufficiently accurate estimates of SVOC concentrations in air so that they can be deployed together with AAS in regional and global monitoring networks.

Knock-Out of Retrovirus Receptor Gene Tva in the Chicken Confers Resistance to Avian Leukosis Virus Subgroups A and K and Affects Cobalamin (Vitamin B12)-Dependent Level of Methylmalonic Acid.

The chicken Tva cell surface protein, a member of the low-density lipoprotein receptor family, has been identified as an entry receptor for avian leukosis virus of classic subgroup A and newly emerging subgroup K. Because both viruses represent an important concern for the poultry industry, we introduced a frame-shifting deletion into the chicken tva locus with the aim of knocking-out Tva expression and creating a virus-resistant chicken line. The tva knock-out was prepared by CRISPR/Cas9 gene editing in chicken primordial germ cells and orthotopic transplantation of edited cells into the testes of sterilized recipient roosters. The resulting tva -/- chickens tested fully resistant to avian leukosis virus subgroups A and K, both in in vitro and in vivo assays, in contrast to their susceptible tva +/+ and tva +/- siblings. We also found a specific disorder of the cobalamin/vitamin B12 metabolism in the tva knock-out chickens, which is in accordance with the recently recognized physiological function of Tva as a receptor for cobalamin in complex with transcobalamin transporter. Last but not least, we bring a new example of the de novo resistance created by CRISPR/Cas9 editing of pathogen dependence genes in farm animals and, furthermore, a new example of gene editing in chicken.

A comprehensive assessment of endocrine-disrupting chemicals in an Indian food basket: Levels, dietary intakes, and comparison with European data.

Endocrine-disrupting chemicals (EDCs) in diet are a health concern and their monitoring in food has been introduced in the European Union. In developing countries, EDC dietary exposure data are scarce, especially from areas perceived as pollution hotspots, including industrialized countries like India. Several persistent organic pollutants (POPs) act as EDCs and pose a pressure to human health mainly through dietary exposure. In the present study, a range of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), dioxins and furans were measured in several food items collected from Indian urban (Delhi) and peri-urban (Dehradun) areas. Food basket contamination data were used to estimate EDC dietary exposure and compare it with that of the average European population estimated from available monitoring data. All the target contaminants were found in most food items, especially in dairies and meat products. OCPs were the main contributers to the measured EDC contamination. Food supplied to Delhi's markets had higher EDC contamination than that supplied to the peri-urban market in Dehradun. Despite lax compliance and control measures, Indian dietary exposure of OCPs and PBDEs were comparable with that of Europe and were lower for PCBs and dioxins. Higher meat consumption in Europe only partly explained this pattern which was driven also by the higher EDC residues in some European food items. A substantial part of endocrine disrupting potential in the diet derives from food and animal feeds internationally traded between developed and developing countries. With increasingly globalized food systems, internationally harmonized policies on EDC content in food can lead to better protection of health in both these contexts.

Predictive modelling as a tool for effective municipal waste management policy at different territorial levels.

Nowadays, the European municipal waste management policy based on the circular economy paradigm demands the closing of material and financial loops at all territorial levels of public administration. The effective planning of treatment capacities (especially sorting plants, recycling, and energy recovery facilities) and municipal waste management policy requires an accurate prognosis of municipal waste generation, and therefore, the knowledge of behavioral, socio-economic, and demographic factors influencing the waste management (and recycling) behavior of households, and other municipal waste producers. To enable public bodies at different territorial levels to undertake an effective action resulting in circular economy we evaluated various factors influencing the generation of municipal waste fractions at regional, micro-regional and municipal level in the Czech Republic. Principal components were used as input for traditional models (multivariable linear regression, generalized linear model) as well as tree-based machine learning models (regression trees, random forest, gradient boosted regression trees). Study results suggest that the linear regression model usually offers a good trade-off between model accuracy and interpretability. When the most important goal of the prediction is supposed to be accuracy, the random forest is generally the best choice. The quality of developed models depends mostly on the chosen territorial level and municipal waste fraction. The performance of these models deteriorates significantly for lower territorial levels because of worse data quality and bigger variability. Only the age structure seems to be important across territorial levels and municipal waste fractions. Nevertheless, also other factors are of high significance in explaining the generation of municipal waste fractions at different territorial levels (e.g. number of economic subjects, expenditures, population density and the level of education). Therefore, there is not one single effective public policy dealing with circular economy strategy that fits all territorial levels. Public representatives should focus on policies effective at specific territorial level. However, performance of the models is poor for lower territorial levels (municipality and micro-regions). Thus, results for municipalities and micro-regions are weak and should be treated as such.

Towards guidelines for time-trend reviews examining temporal variability in human biomonitoring data of pollutants.

In the last few decades, a plethora of studies have focused on human biomonitoring (HBM) of chemical pollutants. Reviewing the copious HBM data reported in these studies is essential for evaluating the effectiveness of pollution management efforts, for example by evaluating time-trends. Nevertheless, guidance to systematically evaluate time trends in published HBM data has never been developed. In this study, we therefore present a proposal for guidelines to conduct "time-trend reviews" (TTRs) that examine time trends in published large HBM datasets of chemical pollutant concentrations. We also demonstrate the applicability of these guidelines through a case study that assesses time-trends in global and regional HBM data on mercury. The recommended TTR guidelines in this study are divided into seven steps: formulating the objective of the TTR, setting up of eligibility criteria, defining search strategy and screening of literature, screening results of search, extracting data, analysing data, and assessing certainty, including the potential for bias in the evidence base. The TTR guidelines proposed in this study are straightforward and less complex than those for conducting systematic reviews assessing datasets on potential human health effects of exposure to pollutants or medical interventions. These proposed guidelines are intended to enable the credible, transparent, and reproducible conduct of TTRs.

Temporal Trends of Persistent Organic Pollutants across Africa after a Decade of MONET Passive Air Sampling.

The Global Monitoring Plan of the Stockholm Convention on Persistent Organic Pollutants (POPs) was established to generate long-term data necessary for evaluating the effectiveness of regulatory measures at a global scale. After a decade of passive air monitoring (2008-2019), MONET is the first network to produce sufficient data for the analysis of long-term temporal trends of POPs in the African atmosphere. This study reports concentrations of 20 POPs (aldrin, chlordane, chlordecone, DDT, dieldrin, endrin, endosulfan, HBCDD, HCB, HCHs, heptachlor, hexabromobiphenyl, mirex, PBDEs, PCBs, PCDDs, PCDFs, PeCB, PFOA, and PFOS) monitored in 9 countries (Congo, Ghana, Ethiopia, Kenya, Mali, Mauritius, Morocco, Nigeria, and Sudan). As of January 1, 2019, concentrations were in the following ranges (pg/m3): 0.5-37.7 (∑6PCB), 0.006-0.724 (∑17PCDD/F), 0.05-5.5 (∑9PBDE), 0.6-11.3 (BDE 209), 0.1-1.8 (∑3HBCDD), 1.8-138 (∑6DDT), 0.1-24.3 (∑3endosulfan), 0.6-14.6 (∑4HCH), 9.1-26.4 (HCB), 13.8-18.2 (PeCB). Temporal trends indicate that concentrations of many POPs (PCBs, DDT, HCHs, endosulfan) have declined significantly over the past 10 years, though the rate was slow at some sites. Concentrations of other POPs such as PCDD/Fs and PBDEs have not changed significantly over the past decade and are in fact increasing at some sites, attributed to the prevalence of open burning of waste (particularly e-waste) across Africa. Modeled airflow back-trajectories suggest that the elevated concentrations at some sites are primarily due to sustained local emissions, while the low concentrations measured at Mt. Kenya represent the continental background level and are primarily influenced by long-range transport.

Precise CRISPR/Cas9 editing of the NHE1 gene renders chickens resistant to the J subgroup of avian leukosis virus.

Avian leukosis virus subgroup J (ALV-J) is an important concern for the poultry industry. Replication of ALV-J depends on a functional cellular receptor, the chicken Na+/H+ exchanger type 1 (chNHE1). Tryptophan residue number 38 of chNHE1 (W38) in the extracellular portion of this molecule is a critical amino acid for virus entry. We describe a CRISPR/Cas9-mediated deletion of W38 in chicken primordial germ cells and the successful production of the gene-edited birds. The resistance to ALV-J was examined both in vitro and in vivo, and the ΔW38 homozygous chickens tested ALV-J-resistant, in contrast to ΔW38 heterozygotes and wild-type birds, which were ALV-J-susceptible. Deletion of W38 did not manifest any visible side effect. Our data clearly demonstrate the antiviral resistance conferred by precise CRISPR/Cas9 gene editing in the chicken. Furthermore, our highly efficient CRISPR/Cas9 gene editing in primordial germ cells represents a substantial addition to genotechnology in the chicken, an important food source and research model.

Comparability of long-term temporal trends of POPs from co-located active and passive air monitoring networks in Europe.

The comparability of data from active (ACT) and passive sampling (PAS) of persistent organic pollutants (POPs) in air is hindered by uncertainties related to the derivation of sampling rates and concentrations, as well as differences in the duration, volume and frequency of sampling. Although data from ACT have been used extensively in short-term PAS calibration studies, no attempts have been made to evaluate the comparability of long-term trends calculated from PAS to established ACT trends. This is crucial, as continuous long-term ACT is unfeasible in most regions of the world. To address these challenges, we calculated and compared trends for organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) at the six sites in Europe with at least 5 years of co-located ACT and PAS data (2012-2016): Birkenes, Kosetice, Pallas, Råö, Stórhöfði and Zeppelin. Strong agreement of ACT and PAS trends was observed for most OCPs and PCBs. Apart from two PCBs at Stórhöfði, all pairs of ACT and PAS trends followed the same direction. However, differences in the magnitude, significance and confidence intervals of their slopes were observed for some compounds and were primarily attributed to the short duration of the PAS time series. Despite some limitations, our results suggest that the comparability of ACT and PAS POP trends will continue to improve with additional years of data. This study confirms the suitability of PAS for the calculation of long-term POP trends in air, and highlights the importance of continuous sampling at established monitoring sites with consistent analytical methods.

Conservation of chicken male germline by orthotopic transplantation of primordial germ cells from genetically distant donors†.

Successful derivation and cultivation of primordial germ cells (PGCs) opened the way to efficient transgenesis and genome editing in the chicken. Furthermore, implantation of male PGCs from non-chicken galliform species into the chicken embryos resulted in cross-species germline chimeras and viable offspring. We have recently improved the PGC technology by demonstrating that chicken male PGCs transplanted into the testes of adult cockerel recipients mature into functional sperms. However, the availability of this orthotopic transplantation for cross-species transfer remains to be explored. Here we tested the capacity of genetically distant male PGCs to mature in the microenvironment of adult testes. We derived PGCs from the Chinese black-bone Silkie and transplanted them into infertile White Leghorn cockerels. Within 15-18 weeks after transplantation, we observed restoration of spermatogenesis in recipient cockerels and production of healthy progeny derived from the transplanted PGCs. Our findings also indicate the possibility of cross-species orthotopic transplantation of PGCs. Thus, our results might contribute to the preservation of endangered avian species and maintaining the genetic variability of the domestic chicken.