Hazardous chemicals in outdoor and indoor surfaces: artificial turf and laminate flooring.

BACKGROUND: Synthetic materials, increasingly used for indoor and outdoor surfaces including homes and playgrounds, may contain toxic chemicals. Infants have a higher potential of exposure to chemicals in these materials, which may pose a risk to their health. OBJECTIVE: To understand potential risks related to outdoor surface coverings, based on a review of the literature and regulations, and to assess levels of hazardous chemicals in surface coverings in Israel. METHODS: We reviewed the literature and regulations on artificial turf. We tested 46 samples of surfaces for trace metals in synthetic playground surfaces; trace metals, phthalates, and di(2-ethylhexyl) terephthalate (DEHT) in synthetic grass, and phthalates, DEHT and formaldehyde in laminate flooring. RESULTS: Twelve studies reporting high levels of polycyclic aromatic hydrocarbons (PAH), and varying levels of trace metals in synthetic playground surfaces were identified, as well as five international regulations on lead with maximum acceptable concentrations in the range 40-500 mg/kg. Surface tests showed that 20 out of 30 samples of synthetic playground surfaces exceeded relevant standards for trace metals, of which five had cadmium levels ≥30 mg/kg and four had chromium levels ≥510 mg/kg. In synthetic grass, three out of eight samples exceeded relevant standards, with lead levels ≥1200 mg/kg. In Laminate flooring (n = 8) formaldehyde levels were in the range of 0.7-1.2 mg/m2 formaldehyde, and five samples contained ~5% DEHT. SIGNIFICANCE: The literature on chemicals in surfaces is limited, but indicates some exceedance of regulatory limits. Trace metals in synthetic playground surfaces and synthetic grass, not regulated in Israel, exceeded relevant international standards in 72% of samples. Laminate flooring, regulated for formaldehyde, did not exceed the 3.5 mg/m2 standard, but contained DEHT, a replacement for ortho-substituted phthalates. The results of this preliminary study show that flooring surfaces may be a source of children's exposure to toxic chemicals. IMPACT STATEMENT: Synthetic surfaces are increasingly being used in, for example, children's playgrounds and sports fields. Exceedances of regulatory limits from other jurisdictions, of heavy metal levels in most outdoor surfaces sampled in Israel indicates the potential for children's exposure. Domestic regulations should be implemented to reduce the risk to children from exposure to these surfaces.

Lead in children's jewelry: the impact of regulation.

BACKGROUND: In 2016 we identified a regulatory gap in Israel reflecting a lack of restrictions on lead in children's jewelry. We conducted surveys that found high levels of lead in children's jewelry. Following the findings, a new standard restricting lead content was introduced in 2018. OBJECTIVE: The goal of this study is to assess whether the new standard had an impact on lead concentrations in children's jewelry on the market 4 months after entry into force, and to examine factors that influenced the standard's effectiveness. METHODS: Thirty-five items of children's jewelry were sampled from stores in Israel in 2018. Lead content of 130 subsamples of these items was tested with X-ray Fluorescence Spectrometry (XRF). We compared these findings to those before the introduction of the standard. We also conducted five structured interviews with professionals from government ministries, industry and expert bodies to examine factors influencing the standard's enactment and implementation. RESULTS: 17% of jewelry samples exceeded the ASTM lead standard in 2018 compared with 50% in 2016. The mean of the jewelry subsamples analyzed in 2018 was 936 (±1700) compared to 1420 (±5740) ppm in 2016. Scientific and regulatory consensus among those setting the new standard led to its swift enactment. However, enforcement challenges may reduce the standard's impact. SIGNIFICANCE: A new standard on lead in children's jewelry was followed by a decline in lead concentrations, but some products with lead exceedances remained available on the market. Public health campaigns, standards and compliance monitoring, and higher penalties will all help to reach the goal of protecting public health.

Nationwide seroprevalence of antibodies against SARS-CoV-2 in Israel.

The first local spread of COVID-19 in Israel was detected in March 2020. Due to the diversity in clinical presentations of COVID-19, diagnosis by RT-PCR alone might miss patients with mild or no symptoms. Serology testing may better evaluate the actual magnitude of the spread of infection in the population. This is the first nationwide seroprevalence study conducted in Israel. It is one of the most widespread to be conducted thus far, and the largest per-country population size. The survey was conducted between June 28 and September 14, 2020 and included 54,357 patients who arrived at the Health Maintenance Organizations to undergo a blood test for any reason. A patient was considered seropositive after two consecutive positive results with two different kits (Abbott and DiaSorin).The overall seroprevalence was 3.8% (95%CI 3.7-4.0), males higher than females [4.9% (95%CI 4.6-5.2) vs. 3.1% (95%CI 2.9-3.3) respectively]. Adolescents had the highest prevalence [7.8% (95%CI 7.0-8.6)] compared to other age groups. Participants who had undergone RT-PCR testing had a tenfold higher risk to be seropositive. The prevalence-to-incidence ratio was 4.5-15.7. Serology testing is an important complimentary tool for assessing the actual magnitude of infection and thus essential for implementing policy measures to control the pandemic. A positive serology test result was recently accepted in Israel as being sufficient to define recovery, with possible far-reaching consequences, such as the deploying of employees to ensure the maintenance of a functional economy.

Multi-center nationwide comparison of seven serology assays reveals a SARS-CoV-2 non-responding seronegative subpopulation.

BACKGROUND: An Israeli national taskforce performed a multi-center clinical and analytical validation of seven serology assays to determine their utility and limitations for SARS-CoV-2 diagnosis. METHODS: Serology assays from Roche, Abbott, Diasorin, BioMerieux, Beckman-Coulter, Siemens, and an in-house RBD ELISA were included. Negative samples from 2391 individuals representative of the Israeli population, and 698 SARS-CoV-2 PCR positive patients, collected between March and May 2020, were analyzed. FINDINGS: Immunoassays sensitivities between 81.5%-89.4% and specificities between 97.7%-100% resulted in a profound impact on the expected Positive Predictive Value (PPV) in low (<15%) prevalence scenarios. No meaningful increase was detected in the false positive rate in children compared to adults. A positive correlation between disease severity and antibody titers, and no decrease in antibody titers in the first 8 weeks after PCR positivity was observed. We identified a subgroup of symptomatic SARS-CoV-2 positive patients (~5% of patients), who remained seronegative across a wide range of antigens, isotypes, and technologies. INTERPRETATION: The commercially available automated immunoassays exhibit significant differences in performance and expected PPV in low prevalence scenarios. The low false-positivity rate in under 20's suggests that cross-reactive immunity from previous CoV strains is unlikely to explain the milder disease course in children. Finding no decrease in antibody titers in the first 8 weeks is in contrast to some reports of short half-life for SARS-CoV-2 antibodies. The ~5% who were seronegative non-responders, using multiple assays in a population-wide manner, represents the proportion of patients that may be at risk for re-infection. FUNDING: Israel Ministry of Health.

Lead in spray paint and painted surfaces in playgrounds and public areas in Israel: Results of a pilot study.

Exposure to low levels of lead in children can cause cognitive deficits (reduced IQ) and behavioral changes such as reduced attention span and increased antisocial behavior. There are no mandatory limits on lead in paint in Israel, with the exception of paints used on toys and children's furniture. However, paints in playgrounds and public areas may be a source of exposure to lead in young children. In this study, we quantified lead concentrations in painted surfaces in public playgrounds and public areas, and in spray paints in Israel, using X-ray fluorescence (XRF). Lead was detected in 43/48 (90%) of surfaces; concentrations exceeded the US regulatory limit for lead in paint (90 ppm) in 4/5 (80%) of tested surfaces on wooden picnic tables and benches, and 22/25 (88%) of surfaces in playgrounds. In 15 surfaces, lead concentrations were 10-700 times higher than the US limit. Out of 11 spray paints sampled, three (27%) had concentrations above the US standard for residential paints. In order to prevent exposure of the general public to lead in playgrounds and public areas and in spray paints, we recommend an integrated approach, including regulatory restrictions (mandatory lead limit of 90 ppm in all paints except labeled industrial paints); awareness raising in the public and in national agencies and local authorities responsible for maintenance in playgrounds, schools and public areas; and encouragement of voluntary measures by industry to prevent sale of industrial paints for use in areas accessible to the public.

Concentrations of trace metals, phthalates, bisphenol A and flame-retardants in toys and other children's products in Israel.

Trace metals and synthetic chemicals including phthalates, bisphenol A and flame retardants, are widely used in toys and childcare products, and may pose acute or chronic adverse health effects in children. In Israel, certain chemicals are regulated in childcare products, but there are still regulatory gaps. We tested regulated and unregulated contaminants in 174 item parts from 70 childcare items with potentially high oral or dermal exposure, including 22 children's jewelry items, 14 toys, 7 diaper-changing mats, 6 baby mattresses, 7 baby textiles and 14 feeding and bathing items. In children's jewelry, an unregulated product in Israel, 23% of samples exceeded the US standard for lead. In toys, a regulated product, we did not detect trace metals above the Israeli standard. In textiles, baby mattresses and diaper-changing mats, phthalates exceeded the European Union standard in 14-45% of tests with a mean of 6.74% by mass for diisononyl phthalate, and 1.32% by mass for di(2-ethylhexyl) phthalate. BPA migration exceeded the EU standard in 14-45% of tests with a mean of 1.03 ppm. The flame retardants polybrominated biphenyls, pentabromodiphenyl, octabromodiphenyl ether, tris(2,3-dibromopropyl) phosphate and tris-(aziridinyl)-phosphine oxide were not detected. For products regulated in Israel, our findings suggest general compliance with mandatory standards. However, a lack of comprehensive chemical regulation means that there are regulatory gaps, and products not regulated in Israel may contain high levels of chemical contamination, exceeding US or EU regulations. The results of this study have prompted the development of an Israeli safety standard for children's jewelry.

Regulation of chemicals in children's products: How U.S. and EU regulation impacts small markets.

Toys and children's products may contain trace metals and organic compounds that are potentially harmful to the health and development of infants and young children. Intergovernmental organizations and individual countries regulate chemicals in consumer products, but a coordinated international approach is lacking. This paper examines the implications of chemical regulation in children's products in large markets for a smaller market, namely Israel. We compared chemical regulations in children's products in the U.S., EU and Israel, and conducted in-depth interviews with diverse stakeholders in the Israeli product standardization process. Israel adopted EU chemical standards for certain chemicals (e.g., trace metals, phthalates) but not others (e.g., bisphenol A, flame retardants, trace metals in children's jewelry). Israeli regulation of chemicals in consumer products relies on regulations in large markets such as the U.S. and EU, which therefore have impacts beyond their territories. However, Israel adopts only product-specific standards and has regulatory gaps due to the lack of an overarching regulatory approach that exists in the U.S. and the EU. Furthermore, Israeli policy is to adopt parallel standards from large markets in order to remove trade barriers, despite their different approaches to chemical regulation, an approach which prioritizes trade considerations over health considerations. We conclude with policy recommendations for Israel, which have relevance for other small markets.

Gender and sexual behavior modulate the composition of serum lipocalins in Nile tilapia (Oreochromis niloticus).

In tilapia species, plasma lipoproteins with high electrophoretic mobility function in intra- and intergender communication. Blood samples taken at onset and peak of daily sexual activity from dominant and subordinate Oreochromis niloticus males and females were fractionated by native gel electrophoresis and the fast-migrating proteins were subjected to mass spectrometry. Mining the sequence data of the Cichlid Genome Consortium, we identified 11 proteins from the lipocalin super-family and characterized their genes' structures. Phylogenetic and structural analyses subdivided these genes into two classes: (I) 3-coding-exon apolipoproteins and (II) more complex 6-coding-exon sulfide-bond-containing lipocalins. Five apolipoproteins and PTGDSL1, TBTBP, and MSP proteins were modulated by gender and sexual behavior. PTGDSL1 protein was only observed in the plasma serum of dominant males. However, the cysteine residue in the position that is crucial for synthetase activity in mammalian prostaglandin D synthetases was not conserved in PTGDSL1 or PTGDSL2 proteins. In line with previous reports suggesting their involvement in male functions as pheromone transporters, TBTBP and MSP proteins were not detected in females at the onset of daily activity. Their increasing amount in males was concordant with the increase in apolipoproteins AFP4L, APOA4a, APOA4b, APO14kD and APOC2, which were detected exclusively in dominant males, indicating a possible role in mobilization of the energy required to maintain their social hierarchy.

Biochemical and in vitro biological significance of natural sequence variation in the ovine leptin gene.

The hormone leptin is involved in diverse biological processes, including regulation of food intake, body-weight homeostasis and energy balance. Sequence variation in the bovine leptin gene has been found to be associated with variations in carcass fat content and average daily gain, as well as in milk yield, milk somatic cell count and several traits governing reproduction. We sequenced genomic DNA and cDNA samples of individuals from three divergent sheep breeds and revealed synonymous as well as novel non-synonymous allelic variation at the third exon of the ovine leptin gene (oLEP) as compared to the sequence published at Accession No. U84247 (reference sequence). In addition, two alternatively spliced oLEP transcripts were found in the abdominal fat tissue. The biochemical and the in vitro biological significance of the sequence variation in the oLEP was examined by generating recombinant oLEP-protein variants namely: p.Q28del, p.N78S, p.R84Q, p.P99Q, p.V123L and p.R138Q, carrying the corresponding sequence variation. Surface plasmon resonance experiments revealed, in most cases, reduced affinity of the oLEP protein variants examined, to human leptin-binding domain (hLBD), relative to the reference variant, being 0.75, 0.60, 0.60, 0.89, 0.92 and 1.03, respectively. In competitive binding assays between biotinylated oLEP and the recombinant leptin protein variants, p.N78S and p.R84Q variants exhibited the lowest affinity to hLBD (0.18 and 0.41, respectively) as compared to the reference hormone. We then tested the protein variants' ability to induce proliferation in Baf-3 cells stably expressing the long form of the human leptin receptor: significant differences in proliferative activity were only found for p.N78S (1.8-fold higher) and p.R138Q (4.2-fold lower) relative to the reference oLEP variant.

A novel day blindness in sheep: epidemiological, behavioural, electrophysiological and histopathological studies.

Four genetically related Improved Awassi sheep flocks had sporadic births of lambs with congenital visual impairments that differed from other known forms of sheep blindness. Pedigree analysis suggested an autosomal recessive mode of inheritance. Behavioural studies of 4-month old affected lambs showed that their day vision (but not night vision) was impaired. Electrophysiological results at this age demonstrated diminished function of cones but not rods. Histopathological and immunohistochemical evaluation of affected retinas from 5-month old lambs revealed both red-green and blue cones, suggesting that the behavioural day blindness and reduced cone electroretinograms reflect cone dysfunction rather than severe cone photoreceptor loss. Awassi day blindness may be a form of achromatopsia.

A mutation in gene CNGA3 is associated with day blindness in sheep.

Lambs with congenital day blindness show diminished cone function, which is characteristic of achromatopsia, a congenital disorder described in humans and dogs. To identify gene(s) associated with sheep day blindness, we investigated mutations in the CNGA3, CNGB3, and GNAT2 genes which have been associated with achromatopsia. Sequencing the coding regions of those genes from four affected and eight non-affected lambs showed that all affected lambs were homozygous for a mutation in the CNGA3 gene that changes amino acid R236 to a stop codon. By PCR-RFLP-based testing, homozygosity for the stop codon mutation was detected in another 19 affected lambs. Non-affected individuals (n=386) were non-carriers or heterozygous for the mutation. While a selection program has been launched to eradicate the day blindness mutation from Improved Awassi flocks, a breeding nucleus of day-blind sheep has been established to serve as animal models for studying human achromatopsia.

Pegylated leptin antagonist is a potent orexigenic agent: preparation and mechanism of activity.

Leptin, a pleiotropic adipokine, is a central regulator of appetite and weight and a key immunomodulatory protein. Although inborn leptin deficiency causes weight gain, it is unclear whether induced leptin deficiency in adult wild-type animals would be orexigenic. Previous work with a potent competitive leptin antagonist did not induce a true metabolic state of leptin deficiency in mice because of a short circulating half-life. In this study, we increased the half-life of the leptin antagonist by pegylation, which resulted in significantly increased bioavailability and retaining of antagonistic activity. Mice administered the pegylated antagonist showed a rapid and dramatic increase in food intake with weight gain. Resulting fat was confined to the mesenteric region with no accumulation in the liver. Serum cholesterol, triglyceride, and hepatic aminotransferases remained unaffected. Weight changes were reversible on cessation of leptin antagonist treatment. The mechanism of severe central leptin deficiency was found to be primarily caused by blockade of transport of circulating leptin across the blood-brain barrier with antagonisms at the arcuate nucleus playing a more minor role. Altogether we introduce a novel compound that induces central and peripheral leptin deficiency. This compound should be useful in exploring the involvement of leptin in metabolic and immune processes and could serve as a therapeutic for the treatment of cachexia.

Prolificacy and lamb survival at birth in Awassi and Assaf sheep carrying the FecB (Booroola) mutation.

Prolificacy up to the fifth parity and lamb survival at birth were investigated in >or=31/32 Awassi and >or=31/32 Assaf sheep belonging to the ++, B+ and BB genotypes at the FecB locus. In the Awassi, prolificacy of ++, B+ and BB ewes was 1.28, 1.90 and 1.92 lambs born/lambing (LB/L), respectively. In the Assaf, prolificacy of ++, B+ and BB ewes was 1.68, 2.40 and 2.55LB/L, respectively. Lamb survival at birth in the ++ Awassi and the ++ Assaf averaged 0.98 and 0.94, respectively. It declined to 0.93 and 0.86, and 0.85 and 0.78 in the B+ and BB Awassi and B+ and BB Assaf, respectively. For singles, twins, triplets, quadruplets and quintuplets, lamb survival rate at birth was 0.98, 0.92, 0.86, 0.78 and 0.65, respectively. FecB genotype-litter size interactions were not significant (P<0.05). A compilation of study results in which prolificacy of the ++ and B+ genotypes at the FecB locus were investigated in a range of breed-environment combinations revealed that the B allele has a multiplicative effect on prolificacy as B+ prolificacy was significantly (P<0.05) linearly associated with ++ prolificacy.

Preparation and characterization of recombinant chicken growth hormone (chGH) and its putative antagonist chGH G119R mutein.

Synthetic cDNA of chicken GH (chGH) and its G119R mutein was synthesized after being optimized for expression in E. coli. The respective cDNAs were inserted into expression vector, expressed and found almost entirely in the insoluble inclusion bodies (IBs). The IBs were isolated, the proteins solubilized in 4.5 M urea, at pH 11.3 in presence of cysteine, refolded, and purified to homogeneity by anion-exchange chromatography on Q-Sepharose. The overall yields were 400 to 500 mg from 5 L of fermentation. Both proteins were > 98% pure, as evidenced by SDS-PAGE, and contained at least 95% monomers, as documented by gel-filtration chromatography under non-denaturing conditions. Circular dichroism analysis revealed that both proteins have identical secondary structure characteristic of cytokines, namely > 50% of alpha helix content. Chicken GH was capable of forming a 1:2 complex with recombinant oGH receptor extracellular domain, but its affinity, as determined by RRA, was 11-fold lower than that of ovine GH (oGH). Correspondingly, its bioactivity, assessed using FDC-P1 3B9 cells stably transfected with rabbit GHR, was 30-40-fold lower, whereas chGH G119R mutant did not bind to oGHR-ECD and was devoid of any biological activity in FDC-P1 3B9 cells. However, in binding experiments that were carried out using chicken liver membranes, both oGH and chGH showed similar IC(50) values in competition with (125)I-oGH, while the IC(50) of G119R mutein was 10-fold higher. These results emphasize the importance of species specificity and indicate the possibility of antagonistic activity of chGH G119R.