Regulation of Persistent Chemicals in Hazardous Waste: A Case Study of Washington State, USA.

Despite ongoing controversy, several strategic frameworks for defining chemicals of concern (e.g., persistent, bioaccumulative, toxic [PBT]; persistent, mobile, toxic [PMT]; persistent organic pollutant [POP]) share persistence as a key criterion. Persistence should be considered over the entire chemical life cycle from production to disposal, including hazardous waste management. As a case study, we evaluate persistence criteria in hazardous waste regulations in Washington state, USA, illustrate impacts on reported waste, and propose refinements in these criteria. Although Washington state defines persistence based on half-life (>1 y) and specific chemical groups that exceed summed concentration thresholds in waste (i.e., >0.01% halogenated organic compounds [HOCs] and >1.0% polycyclic aromatic hydrocarbons [PAHs]), persistence is typically addressed with HOC and PAH evaluation but seldom with half-life estimation. Notably, persistence is considered (with no specific criteria) in corresponding federal regulations in the United States (Resource Conservation and Recovery Act). Consequently, businesses in Washington state report annual amounts of state hazardous waste (including persistent waste) separately from federal hazardous waste. Total state-only waste, and total state and federal waste combined, nearly doubled (by weight) from 2008 to 2018. For the period 2016 to 2018, persistence criteria captured 17% of state-only waste and 2% of total state and federal waste combined. Two recommendations are proposed to improve persistence criteria in hazardous waste regulations. First, Washington state should consider aligning its half-life criterion with federal and European Union PBT definitions (e.g., 60-120 d) for consistency and provide specific methods for half-life estimation. Second, the state should consider expanding its list of persistent chemical groups (e.g., siloxanes, organometallics) with protective concentration thresholds. Ultimately, to the extent possible, Washington state should strive toward harmonizing persistence in hazardous waste regulations with corresponding criteria in global PBT, PMT, and POP frameworks. Integr Environ Assess Manag 2021;17:455-464. © 2020 SETAC.

Experimental IgA nephropathy.

An animal model for IgA immune complex nephritis was developed. IgA immune complexes formed in vitro with an IgA anti-dinitrophenyl (DNP) derived from MOPC-315 plasmacytoma, and dinitrophenylated bovine serum albumin (DNP-BSA) produced mild focal glomerulonephritis in mice. Similar, but more severe pathological changes were produced with complexes formed in vivo either in normal mice or MOPC-315 tumor-bearing mice. In contrast to the focal nature of the PAS-positive glomerular lesions observed by light microscopy, immunofluorescent examination revealed IgA deposits in all glomeruli. This discrepancy between immunofluorescent and histopathologic findings as well as the distribution of the immune complexes within the affected glomeruli, are some of the features which bear resemblance between this experimental model and human IgA nephropathy. Fixation of complements by DNP-BSA-IgA immune complexes, formed in vitro or in vivo, was shown to occur in the glomeruli of mice with IgA immune complex nephropathy. The pattern of C3 glomerular deposits was similar to that of IgA. However, complement proved to be nonessential for complex deposition. This conclusion is based on the observation that decomplemented mice, although showing no deposition of C3 in their glomerulus, developed glomerular immunohistological changes similar to those observed in experimental mice that were not decomplemented. Polymeric IgA was observed to be critical for renal deposition of complexes and induction of nephritic histological changes. In contrast, monomeric IgA immune complexes failed to produce glomerular deposits. This finding raises the possibility that secretory IgA, which is predominantly polymeric, may play a role in human IgA-associated glomerulonephritis.

Differences in the localization and morphology of chromosomes in the human nucleus.

Using fluorescence in situ hybridization we show striking differences in nuclear position, chromosome morphology, and interactions with nuclear substructure for human chromosomes 18 and 19. Human chromosome 19 is shown to adopt a more internal position in the nucleus than chromosome 18 and to be more extensively associated with the nuclear matrix. The more peripheral localization of chromosome 18 is established early in the cell cycle and is maintained thereafter. We show that the preferential localization of chromosomes 18 and 19 in the nucleus is reflected in the orientation of translocation chromosomes in the nucleus. Lastly, we show that the inhibition of transcription can have gross, but reversible, effects on chromosome architecture. Our data demonstrate that the distribution of genomic sequences between chromosomes has implications for nuclear structure and we discuss our findings in relation to a model of the human nucleus that is functionally compartmentalized.

Analysis of two purified mutants of Escherichia coli aspartate transcarbamylase with single amino acid substitutions.

Two mutant versions of Escherichia coli aspartate transcarbamylase have been purified and analyzed kinetically. Each of these mutant enzymes contains a single amino acid different from the wild-type enzyme, which was introduced by suppression of a nonsense codon within the E. coli pyrB gene. These enzymes exhibited alterations in both homotropic and heterotropic interactions with little change in specific activity. Depending upon the site of the substitution, the allosteric interactions have been either enhanced or diminished over the wild-type enzyme. Carbamyl phosphate saturation curves indicate that aspartate and carbamyl phosphate homotropic co-operativity are separable. Experiments employing the allosteric effectors indicate that the transmission of the regulatory effect is dependent upon the structure of the catalytic subunit, and that CTP inhibition can be partially decoupled from ATP activation. The kinetics of one of the mutants is unusually sensitive to dissociation at elevated temperatures. This sensitivity may be due to weakened interactions between the subunits of the enzyme.

Thyroid antigen associated immune complex glomerulonephritis in Graves' disease.

A 60 year old hyperthyroid black woman with long-standing Graves' disease treated with methimazole presented with anasarca and congestive heart failure. She was found to have the nephrotic syndrome with a urinary protein excretion of 32 g/day. Light and electron microscopy revealed a stage II membranous glomerulopathy. Direct and indirect immunofluorescence demonstrated immunoglobulin G (IgG), immunoglobulin M (IgM), third component of complement and thyroglobulin in a granular diffuse pattern consistent with an immune complex glomerulonephritis. Total thyroidectomy led to a decrease in proteinuria with little change in glomerular filtration rate during an 11 month follow-up period. We believe this to be the first report of immune complex glomerulonephritis associated with thyroid antigen in Graves' disease.

Psychiatric disorder and cognitive function in a family with an inherited novel mutation of the developmental control gene PAX6.

The PAX family of developmental control genes are known to play important roles in the early patterning of the central nervous system. One member of this family, PAX6, is involved in eye development in invertebrates as well as in mouse and man, but is also widely expressed in the developing forebrain. Humans with a mutation in this gene have abnormalities of eye development, and the results presented here suggest, for the first time, that this mutation may also be associated with subtle abnormalities of frontal lobe function in the family studied. We carried out genotyping of individuals within a single family, with and without the characteristic eye abnormalities of PAX6 mutation, and only those individuals with the mutation showed significant abnormalities on tests of frontal lobe function. These individuals also had higher rates of psychiatric disorder. PAX6 is highly conserved between mouse and man, and although the neuroanatomical phenotype associated with PAX6 heterozygosity has only been studied in mice, the resultant cellular disorganization seen in mice is likely to be present in the human forebrain. Although these mice have no obvious behavioural phenotype, the results presented here suggest that humans with the equivalent mutation display a neurobehavioural phenotype.

Recombination patterns around the breakpoints of a balanced 1;11 autosomal translocation associated with major mental illness.

The frequency and extent of pairing failure around human translocations are unknown. We have examined the pattern of recombination around the breakpoints of a balanced autosomal translocation t(1;11) (q43;q21) associated with major mental illness. DNA was available from 17 carriers and 10 non-translocation carriers with meioses involving four generations. The derivative 1 and 11 chromosomes were also isolated in somatic cell hybrids and used to confirm phase. We have genotyped pedigree members using 20 polymorphic markers within 10 cM on either side of both chromosome 1 and 11 breakpoints. We find no significant reduction of recombination in the vicinity of either breakpoint. However we estimate that there are insufficient meioses even in this large family to make a meaningful interpretation and suggest that sperm typing alone can answer these interesting questions.

Identification of polymorphisms within Disrupted in Schizophrenia 1 and Disrupted in Schizophrenia 2, and an investigation of their association with schizophrenia and bipolar affective disorder.

We have undertaken a search for polymorphic sequence variation within Disrupted in Schizophrenia 1 and Disrupted in Schizophrenia 2 (DISC1 and DISC2), which are both novel genes that span a translocation breakpoint strongly associated with schizophrenia and related psychoses in a large Scottish family. A scan of the coding sequence, intron/exon boundaries, and part of the 5' and 3' untranslated regions of DISC1, plus 2.7 kb at the 3' end of DISC2, has revealed a novel microsatellite and 15 novel single nucleotide polymorphisms (SNPs). We have tracked the inheritance of four of the SNPs through multiply affected families, and carried out case-control association studies using the microsatellite and four common SNPs on populations of patients with schizophrenia or bipolar affective disorder versus normal control subjects. Neither co-segregation with disease status nor significant association was detected; however, we could not detect linkage disequilibrium between all these markers in the control population, arguing that an even greater density of informative markers is required to test rigorously for association in this genomic region.

Suture resistance to infection.

The emergence of both absorbable and nonabsorbable synthetic materials has been primary among the many new developments in suture materials. This is a study of 16 types of natural and synthetic suture materials and their resistance to both gram-positive and gram-negative infections. Four hundred and twenty suture inplants were made in the Edlich mouse model. The 16 different sutures were compared to appropriate controls and graded by the degree of infectibility. Synthetic sutures were superior in all areas. The monofilament sutures performed better than the multifilament sutures. Lubricating coatings had no effect on infectibility. Natural sutures performed poorly and should not be used in wounds that are potentially susceptible to infection.

Arterial occlusive disease: a function of vessel bifurcation angle.

A clinical observation that patients with aortoiliac occlusive disease have a high aortic bifurcation serves as the stimulus for this study. A review of 100 consecutive abdominal aortograms revealed that patients with occlusive disease had an average bifurcation angle of 38 degrees and those with normal studies or aneurysmal disease had an average angle of 52 degrees. A flow visualization system was constructed to study shear stress using eight Pyrex models with varying bifurcation angles (20 to 90 degrees). The perfusate consisted of a 35% sucrose solution with anion-exchange beads to serve as flow particles. The bifurcations were photographed at three different flow rates. The length of each tracer particle was measured to determine its velocity. Velocity profile curves were constructed and shear stress calculated by the formula shear stress = mu x dv/dr, where mu is viscosity, v is velocity, and r is radius. Results of the study suggest that more acute angles cause a greater shear stress on the inner wall of the bifurcation and decreased shear stress on the outer wall. Two theories of atheroma predilection, high shear stress and low shear stress, are applied to patients with high acute aortic bifurcations.

Clinical and genetic analysis of a large North European Caucasian family affected by early-onset periodontitis.

Genetic studies of early-onset periodontitis (EOP) are hampered by several factors. These include delayed onset of the trait, an upper age limit of expression of the disease, and lack of phenotypic information for edentulous family members. Segregation analyses of families with EOP support a major locus hypothesis but fail to define clearly the criteria used for diagnosis of the relatives. Confirmation of a proposed mode of inheritance and the identification of risk genes is awaited by means of family linkage studies. It is suggested that a system can be developed for the current and retrospective diagnosis of relatives of EOP probands. In addition, it is hypothesized that the large family presented here is suitable for a linkage study. Relatives of the proband who were unavailable for a full periodontal examination, were edentulous, or were deceased, were diagnosed by means of documented clinical evidence of periodontal disease or from reported case histories. Segregation analysis was performed. Analysis of the power of the pedigree to detect linkage was carried out by means of the SIMLINK program. Three different categories were defined according to the reliability of diagnosis of EOP. Segregation analysis indicated either autosomal-dominant or X-linked-dominant inheritance in this family. The simulations showed lod scores above 3.0 for all locations of the disease gene, and for each category of diagnosis. In conclusion, a method has been developed which can be used for the diagnosis of relatives of EOP probands when ideal clinical data are unavailable. The simulations suggest that this family is suitable for a genetic linkage study with the aim of identifying the location of one or more susceptibility genes.

The effect of dietary fat and protein levels, form and cereal type on fatty liver and kidney syndrome in chicks.

A total of 4200 day-old Ross I broiler chicks were subjected to an ambient temperature 2 to 3C degrees higher than normal and were fed various diets to determine the effects of dietary factors and sex on mortality from fatty liver and kidney syndrome (FLKS). In a first experiment mortality was not significantly influenced by cereal type (barley or wheat) but was influenced significantly by fat content of the diet. In a second experiment mortality was increased significantly with a low protein diet and with a pelleted one compared with a mash diet. In both experiments mortality was signigicantly higher in females than in males. High levels of protein and fat in the diet had a protective influence on mortality but the results, especially with pelleted diets, suggested that some other nutrient(s) might be involved in the syndrome.

Estimating human inbreeding coefficients: comparison of genealogical and marker heterozygosity approaches.

We have used genealogies and genomic polymorphisms to estimate individual inbreeding coefficients (F) in 50 subjects with an expected range (based on recent genealogies) of F from 0.0 to 0.0625. The estimates were based on two approaches, using genotypes respectively from 410 microsatellite markers (410-STR panel) and from 10,000 SNPs (10K-SNP panel). The latter was performed in a sub-sample of 15 individuals. We concluded that for both marker panels measures of inbreeding based on the excess of homozygosity over Hardy-Weinberg expectation were not closely correlated with 4-5 generation genealogical F-values. For the 10K-SNP panel we found two alternative measures which correlated more closely with F, based respectively on standard errors and on paired homozygosity of nearby SNPs over distances of 2-4 cM. We propose an empirical method for estimating standard errors and hence individual F-values, based on the variation between individual autosomes. This method could provide useful estimates of average F-values for groups of individuals in population-based studies of the effects of inbreeding/homozygosity on quantitative traits.

Different responses of two mouse strains to 650 rads and protection by surgical stress.

Radiation survival of C57B1/6J mice after 650 rads improved from 1.7% to 66% when mice were stressed surgically one week before irradiation. Surgical stress one or three weeks before irradiation increased endogenous CFUs and decreased exogenous CFUs in C57B1/6J and DBA/1J mice. Survival may be related to the proportion of stem cells in different phases of the cell cycle, which was abnormal in C57B1/6J mice and compensated for by a greater number of nucleated cells per femur.

Age-associated involution of cellular immune function. I. Accelerated decline of mitogen reactivity of spleen cells in adult thymectomized mice.

Autoimmune-susceptible (A/J and C57BL/6J) and nonautoimmune-susceptible (DBA/1J) strains of mice were adult thymectomized (aTx). Groups of mice treated by aTx, aTx and 330 rads (R), aTx and 650 R, and 330 R OR 650 R only, were compared to normal age-matched control (normal and sham Tx) mice. T cell functions were then followed as measured by in vitro spleen cell responsiveness to the T cell mitogens phytohemagglutinin (PHA) and staphylococcal enterotoxin B (SEB) in a long-term study. A decline in spleen cell responsiveness with natural aging up to 18 months of age was seen to PHA in all three strains and to SEB in A/J and C57BL/J mice. Adult Tx alone was followed by an accelerated decline in spleen cell responsiveness to PHA and SEB in all three strains of mice. Irradiation combined with aTx-reduced spleen cell responsiveness to SEB in A/J and DBA/1J mice. Irradiation with or without aTx diminished the bone marrow stem cell pool as measured by exogenous colony-forming units, and the hematocrit of all three strains of mice. Thus, the natural decline in cellular immunity with aging was accelerated by aTx in both autoimmune-susceptible and non-autoimmune-susceptible strains of mice. These results emphasize that physiologic thymic function(s) must continue throughout life in order to maintain T cell function.

Pyrolytic carbon-coated Dacron grafts.

A new pyrolytic carbon surface (ULTI) was tested on a loosely knitted dacron substrate in dogs. Patency was 97.5% as compared with a control of 33%. Physical properties of the dacron were not altered by the ULTI carbon deposition process. This modified pyrolytic carbon on a dacron substrate is a very promising vascular biomaterial for small vessel replacement.

The adrenal cortico-medullary ratio in the fowl.

1. A new integrating method of assessing the proportions of cortical and medullary tissue in the avian adrenal gland is described and statistically evaluated. 2. It is shown that a single "central" section of the adrenal gland will suffice to obtain accurate results. 3. Each section was projected on to a grid of about 2500 intersection points and the ratio of points falling on cortical or medullary tissue was determined. 4. Applications of this method to normal fowls showed that there were significant breed, sex and, particularly, age differences in the cortico-medullary ratio of the fowl's adrenal glands.