A longitudinal study of piece rate and health: evidence and implications for workers in the US gig economy.

OBJECTIVES: The objective of this study was to evaluate health outcomes for workers subject to piece rate historically to better understand the implications of pay type in the modern-day gig economy. While piece rate occurring in the 1980s and 1990s predates recent platform-based employment, it introduced and normalized patterns of economic precariousness that are instrumental in the current gig economy. Evidence suggests that such pay types may result in poor health outcomes; however, cross-sector evidence of its long-term effects on US workers is lacking. This article represents the first longitudinal cross-sector analysis relating health outcomes to this performance pay type in US workers. STUDY DESIGN: This is a longitudinal cohort study. METHODS: Data from six survey waves of the 1979 National Longitudinal Survey of Youth collected between 1988 and 2000 are used in a random-effects logit model to predict self-reported health limitations related to piece rate, while controlling for worker, work environment, lifestyle, time, and location trends. RESULTS: Pay tied to piece rate in current or prior periods significantly increases the odds of self-reported health limitations compared with salaried work (odds ratio [OR]: 1.4-1.8). These effects are elevated for the subgroups of low-wage (OR: 1.5-1.8), female (OR: 1.8-1.9), and non-white (OR: 2.0-2.1) workers compared with their high-wage, male, and white peers. CONCLUSIONS: The results suggest that piece rate pay designed to promote efficiency may have important negative implications for worker health, especially for the most vulnerable members of the US workforce such as women, minority, and low-income workers. Given the growing popularity of performance-based pay to the gig economy, more research is needed to determine if the practice is justified from a public health perspective.

Associations between genetic variants in the promoter region of the insulin-like growth factor-1 (IGF1) gene and blood serum IGF1 concentration in Hanwoo cattle.

In this study, we investigated the associations between genetic variants in the promoter region of the insulin-like growth factor-1 (IGF1) gene and blood serum IGF1 concentration in Hanwoo cattle. Polymerase chain reaction primers were based on GenBank accession No. AF404761 and amplified approximately 533-bp segments. Newly identified sequences were submitted to GenBank (accession No. DQ267493). Sequence analysis revealed that genetic variants were located at a nucleotide position 323 for the nucleotide substitution C/A that was first reported in this study and positions 326-349 for a repeat motif (CA10-11). The allele frequencies of g.323C>A were 0.264 (C) and 0.736 (A) without significant deviation from Hardy-Weinberg equilibrium. Frequencies of the repeat motif CA(10) and CA(11) were 0.604 and 0.396, respectively. Statistical analysis revealed that the genetic variation g.323C>A was significantly associated with blood serum IGF1 concentrations with significant additive genetic effects, whereas no associations were found for the repeat motif. IGF1 concentrations were positively (r = 0.453) and negatively (r = -0.445) correlated with weights in the growing stages (16-21 months) and late fattening stages (22-30 months), respectively. The results of the present study and future genotypic data for Hanwoo beef cattle based on the robust genetic variation of IGF1 will provide critical information for genetic improvement and will have a large impact on commercial markets.

Effects of calpain genotypes on meat tenderness and carcass traits of Angus bulls.

Relationships of the calpain system with meat tenderness and carcass traits were examined for 94 purebred Angus bulls with genotypes of the calpain classified by RFLP (restriction fragment length polymorphism) and SSCP (Single strand conformation polymorphism) analysis. Designing of primers based on the calpain regulatory subunit (CAPNS) and u-calpian (CAPN1) genes. Bulls from 15 months of age were slaughtered, and carcass traits, including fat thickness (FAT); longissimus muscle area (LMA); percentage of kidney, pelvic, and heart fat (KPH); hot carcass weight (HCW); marbling score (MAR); and quality grade (QUL), were analyzed. Measurements regarding meat tenderness involved activities of calpastatin (CAC), u-calpain (UAC), m-calpain (MAC), Warner-Bratzler Shear Force (WBS) and myofibril fragmentation index (MFI). Statistical significances of the calpain genotypes accounted for variations in MAR and QUL at CAPNS locus, and both loci explained variations of UAC and MAC. Significant mean differences in genotypes of CAPNS locus were found for MAR (BB > AB > AA) and QUL (AB > BB > AA). UAC showed significant correlations with MAC, CAC, MFI, FAT, and MAR, and we found that MAC correlated with WBS, FAT, HCW, MAR, and QUL. Strong positive correlation detected between LMA and HCW, and MAR and QUL, and a negative correlation between MFI and MAR was estimated. From the result it may be possible to use the calpain genotypes classified by RFLP and SSCP analysis in marker assisted selection programs to estimate UAC and MAC precisely regardless meat tenderness and to improve MAR and QUL of beef cattle.

Late glacial stage and holocene tropical ice core records from huascaran, peru.

Two ice cores from the col of Huascarán in the north-central Andes of Peru contain a paleoclimatic history extending well into the Wisconsinan (Würm) Glacial Stage and include evidence of the Younger Dryas cool phase. Glacial stage conditions at high elevations in the tropics appear to have been as much as 8 degrees to 12 degrees C cooler than today, the atmosphere contained about 200 times as much dust, and the Amazon Basin forest cover may have been much less extensive. Differences in both the oxygen isotope ratio zeta(18)O (8 per mil) and the deuterium excess (4.5 per mil) from the Late Glacial Stage to the Holocene are comparable with polar ice core records. These data imply that the tropical Atlantic was possibly 5 degrees to 6 degrees C cooler during the Late Glacial Stage, that the climate was warmest from 8400 to 5200 years before present, and that it cooled gradually, culminating with the Little Ice Age (200 to 500 years before present). A strong warming has dominated the last two centuries.

Holocene--late pleistocene climatic ice core records from qinghai-tibetan plateau.

Three ice cores to bedrock from the Dunde ice cap on the north-central Qinghai-Tibetan Plateau of China provide a detailed record of Holocene and Wisconsin-Würm late glacial stage (LGS) climate changes in the subtropics. The records reveal that LGS conditions were apparently colder, wetter, and dustier than Holocene conditions. The LGS part of the cores is characterized by more negative delta(18)O ratios, increased dust content, decreased soluble aerosol concentrations, and reduced ice crystal sizes than the Holocene part. These changes occurred rapidly approximately 10,000 years ago. In addition, the last 60 years were apparently one of the warmest periods in the entire record, equalling levels of the Holocene maximum between 6000 and 8000 years ago.

SSZ-26 and SSZ-33: Two Molecular Sieves with Intersecting 10- and 12-Ring Pores.

The framework structures of two closely related molecular sieves, SSZ-26 and SSZ-33, are described. These materials possess a previously missing but desired structural feature in a group of industrially significant zeolites. They contain a three-dimensional pore system that provides access to the crystal interior through both 10- and 12-rings. This property is a consequence of the organic structure-directing agents used in the synthesis of these materials. These materials are examples of the purposeful design of a micropore architecture. Both SSZ-26 and SSZ-33 contain the 4=4-1 building unit that had been previously found only in natural zeolites.

Life cycle efficiency of beef production: IX. Relationship between residual feed intake of heifers and cow efficiency ratios based on harvest, carcass, and wholesale cut weight outputs.

Data were collected from 1953 through 1980 from identical and fraternal twin beef and dairy females born in 1953, 1954, 1959, 1964, and 1969, from crossbred females born as singles in 1974, and their progeny. Numbers of dams that weaned at least one calf and were included in the first analysis were 37, 45, and 56 in the 1964, 1969, and 1974 data sets, respectively. Respective numbers of dams that weaned three calves and were included in a second analysis were 6, 8, 8, 22, 33, and 33 in the 1953, 1954, 1959, 1964, 1969, and 1974 experiments. Individual feed consumption was measured at 28-d intervals from the time females were placed on the experiment at 240 d of age until three calves were weaned or the dams had reached 5 yr of age. Residual feed intake (RFI) and residual BW gain (RG) of heifers that subsequently became dams were determined based on ADG and DMI from 240 d of age to first calving. Various measures of cow efficiency were calculated on either a life cycle or actual lifetime basis using ratios of progeny and dam weight outputs to progeny and dam feed inputs. The correlation between RFI and DMI was large and positive (r = 0.67; P < 0.0001), and RG was highly correlated with ADG (r = 0.75; P < 0.0001). Correlations of RFI with cow efficiency ratios that included harvest weight, carcass weight, or weight of trimmed wholesale cuts as measures of output ranged from -0.05 (P > 0.10) to -0.17 (P < 0.10), indicating that heifers with better (i.e., more negative) RFI values tended to become slightly more efficient cows. Correlations of RG with life cycle and actual lifetime cow efficiency ratios ranged from 0.08 (P > 0.10) to 0.23 (P < 0.05), demonstrating that heifers with better (i.e., more positive) values for RG were somewhat more efficient as cows. The correlations were stronger when cow salvage value was included in the measures of cow efficiency. Correlations of DMI and mid-test metabolic BW (MMW) with life cycle cow efficiency ratios that did not include cow salvage value as output ranged from -0.15 (P < 0.10) to -0.22 (P < 0.01). Correlations of DMI and MMW with actual lifetime cow efficiency ratios varied from -0.20 (P < 0.05) to -0.36 (P < 0.001). Therefore, smaller heifers that consumed less feed had superior cow efficiency ratios. Correlations of RFI with carcass grade, backfat thickness, marbling score, and kidney fat of progeny indicated that heifers with superior RFI would tend to produce leaner offspring.

Shear stress regulates endothelial nitric oxide synthase expression through c-Src by divergent signaling pathways.

In this study, we defined the signaling cascade responsible for increased eNOS mRNA expression in response to laminar shear stress. This pathway depends on the tyrosine kinase c-Src because shear induction of eNOS mRNA is blocked by the c-Src inhibitors PP1 and PP2, as well as an adenovirus encoding kinase inactive c-Src. After activation of c-Src, this pathway diverges. One arm is responsible for the short-term (6 hour) increase in eNOS mRNA. This involves a transient, 1-hour increase in eNOS transcription, as detected by nuclear run-on, that is dependent on activation of Ras and is blocked by adenoviral infection with dominant negative Ras. Downstream of Ras, MEK1/2 and ERK1/2 are important in this pathway, as 2 inhibitors of MEK1/2, PD98059 and UO126, completely prevented this early increase in eNOS mRNA. ERK1/2 was rapidly phosphorylated in response to shear, and this was prevented by c-Src and Ras inhibition. Further, Raf is phosphorylated in response to shear stress, and this is prevented by c-Src inhibition, suggesting that Raf may transduce the signal between Ras and ERK1/2. The second arm of the pathway linking activation of c-Src to eNOS expression involves stabilization of eNOS mRNA by shear stress. This response to shear is completely abrogated by the c-Src inhibitor PP1 but not altered by Ras or MEK1/2 inhibition. Thus, c-Src plays a central role in modulation of eNOS expression in response to shear stress via divergent pathways involving a short-term increase in eNOS transcription and a longer-term stabilization of eNOS mRNA.

Transcriptional and posttranscriptional regulation of endothelial nitric oxide synthase expression by hydrogen peroxide.

Diverse stimuli, including shear stress, cyclic strain, oxidized LDL, hyperglycemia, and cell growth, modulate endothelial nitric oxide synthase (eNOS) expression. Although seemingly unrelated, these may all alter cellular redox state, suggesting that reactive oxygen intermediates might modulate eNOS expression. The present study was designed to test this hypothesis. Exposure of bovine aortic endothelial cells for 24 hours to paraquat, a superoxide (O(2)(-*))-generating compound, did not affect eNOS mRNA levels. However, cotreatment with paraquat and either Cu(2+)/Zn(2+) superoxide dismutase or the superoxide dismutase mimetic tetrakis(4-benzoic acid)porphyrin chloride increased eNOS mRNA by 2.3- and 2.2-fold, respectively, implicating a role for H(2)O(2). Direct addition of 100 and 150 micromol/L H(2)O(2) caused increases in bovine aortic endothelial cell eNOS mRNA that were dependent on concentration (ie, 3.1- and 5.2-fold increases) and time, and elevated eNOS protein expression and enzyme activity, accordingly. Nuclear run-on and 5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole-chase studies showed that H(2)O(2) caused a 3.0-fold increase in eNOS gene transcription and a 2.8-fold increase in eNOS mRNA half-life. Induction of eNOS by H(2)O(2) was not affected by the hydroxyl radical scavenger DMSO, mannitol, or N-tert-butyl-alpha-phenylnitrone, but it was inhibited by the antioxidants N-acetylcysteine, ebselen, and exogenously added catalase. Unlike H(2)O(2), the 4.0-fold induction of eNOS by shear stress (15 dyne/cm(2) for 6 hours) was not inhibited by N-acetylcysteine or exogenous catalase. In conclusion, H(2)O(2) increases eNOS expression through transcriptional and post-transcriptional mechanisms. Although H(2)O(2) does not mediate shear-dependent eNOS regulation, it is likely to be involved in regulation of eNOS expression in response to other physiological and/or pathophysiological stimuli.

The influence of different management systems and age on intestinal morphology, immune cell numbers and mucin production from goblet cells in post-weaning pigs.

At time of weaning, the immune system in piglets is not fully mature resulting in reduced growth and increased mortality. Early-weaned pigs transported to a segregated early weaning (SEW) facility have enhanced performance and gut development compared to conventional (CONV) pigs which may be due, in part, to decreased pathogen challenge. To gain further insight into SEW enhanced performance and gut development, gut samples from pigs weaned at 19+/-2 days were assessed during the post-weaning (PW) period. The numbers of cells expressing CD2, CD4, CD8, and CD172 (the 74-22-15 (SWC3) antibody is now known to be specific for CD172), MHC class II, and CD25 were quantified using immunohistochemistry. Additionally, samples of duodenum, jejunum, and ileum were evaluated for the production of neutral, acidic, and sulfuric mucins from goblet cells and morphological measurements were also made. No effects due to the management systems alone were observed for any of the parameters. However, there were interactive effects of age/time post-weaning and management system on the immune cells as well as on the mucin secreting goblet cells. There were no differences in gut morphology between SEW and CONV reared pigs.

Analysis of female reproductive traits in Angus beef cattle divergently selected for blood serum insulin-like growth factor I concentration.

Insulin-like growth factor-I (IGF-I) is an anabolic polypeptide involved in reproductive performance in several species. The objectives of this study were to determine relationships of pregnancy rate, and age of heifers at puberty and at first calving with serum IGF-I concentration in Angus beef cattle. Data were obtained from an ongoing divergent selection experiment for IGF-I concentration involving purebred Angus cows. The IGF-I concentrations measured at Days 28, 42, and 56 of the 140-day postweaning test are abbreviated as IGF28, IGF42, and IGF56, respectively. Pregnancy rate did not differ between high and low IGF-I line females (P=0.95; n=2618), but high line heifers tended to be 4.02+/-2.18 days younger (P=0.07; n=281) at first calving. Residual correlations of age of heifers at first calving (AFC) with IGF-I measurements were not significant. The linear and quadratic terms for regression of AFC on IGF-I concentrations were also non-significant. Contrast analysis showed no difference in age at puberty between the high and low IGF-I line heifers (5.3+/-6.4 days earlier in the high line; P=0.43; n=51). Residual correlations of age of heifers at puberty with IGF28, IGF42, IGF56, and mean IGF-I were -0.30 (P=0.03), -0. 22 (P=0.12), -0.35 (P=0.01), and -0.34 (P=0.01), respectively. The observed relationships between female reproductive traits and IGF-I concentration in Angus beef cattle suggest complex and multiple roles for IGF-I in reproduction.

Life cycle efficiency of beef production: VIII. Relationship between residual feed intake of heifers and subsequent cow efficiency ratios.

Data were collected from 1953 through 1980 from identical and fraternal twin beef and dairy females born in 1953, 1954, 1959, 1964, and 1969, and from crossbred females born as singles in 1974, and their progeny. Numbers of dams that weaned at least 1 calf and were included in the first analysis were 37, 45, and 56 in the 1964, 1969, and 1974 data sets, respectively. Respective numbers of dams that weaned 3 calves and were included in a second analysis were 6, 8, 8, 22, 33, and 33 in the 1953, 1954, 1959, 1964, 1969, and 1974 experiments. Individual feed consumption was measured at 28-d intervals from the time females were placed on the experiment until 3 calves were weaned or the dams had reached 5 yr of age. Residual feed intake (RFI) and residual gain (RG) of the heifers that subsequently became the dams in this study were determined based on ADG and DMI from 240 d of age to first calving. Various measures of cow efficiency were calculated on either a life cycle or actual lifetime basis using ratios of progeny and dam weight outputs to progeny and dam feed inputs. Residual feed intake was phenotypically independent of ADG and metabolic midweight (MMW), whereas the correlation between RFI and DMI was positive and highly significant ( = 0.67; < 0.0001). Residual gain was highly correlated with ADG ( = 0.75; < 0.0001) and had near 0 correlations with DMI and MMW. Correlations indicated that heifers that ate less and had smaller metabolic midweights from 240 d of age to first calving had superior efficiency ratios as cows. Residual feed intake was not significantly correlated with age at puberty, age at calving, or milk production. Results of this study do not indicate any serious antagonisms of postweaning heifer RFI with subsequent cow and progeny performance traits or with life cycle or actual lifetime cow efficiency. In addition, selection for increased RG would result in earlier ages at calving, but would also tend to result in taller and heavier cows.

Combined conformational search and finite-difference Poisson-Boltzmann approach for flexible docking. Application to an operator mutation in the lambda repressor-operator complex.

The N-terminal domain of the phage lambda repressor binds as a dimer to its palindromic DNA operator sequence. In addition to a helix-turn-helix DNA recognition motif, the first six amino acids of the phage lambda repressor form a flexible peptide segment which wraps around DNA. Site-directed mutagenesis studies have shown that amino acid replacements or partial removal of the arm structure, or changes in the DNA sequence contacting the N-terminal arm, can lower the repressor-operator binding affinity by several orders of magnitude. The finite-difference Poisson-Boltzmann approach in combination with a conformational search procedure was used to study energetic contributions of the lambda arm to repressor-operator recognition based on the high resolution X-ray structure. It allows for the local relaxation of the structure upon changing the DNA sequence in the lambda arm binding region. A simplified potential energy function including torsional, truncated Lennard-Jones and approximate electrostatic terms is used in the initial step to screen out energetically unfavorable structures. The electrostatic energy of selected conformations is subsequently calculated more accurately using the finite-difference Poisson-Boltzmann approach. The method was applied to study the effect of a C-->T mutation at position 6 of the consensus half-site of the operator. This base-pair contacts Lys4 which is part of the arm segment. Keeping only the Lys4 side-chain mobile and with the wild-type DNA operator sequence, several conformations close to the X-ray structure were identified as those with lowest energy. In the case of the DNA mutation, lowest energy conformations differed significantly from those selected for the wild-type sequence. These initial calculations indicate that the approach might be a useful tool to estimate conformational and energetic effects upon mutagenesis of protein-DNA complexes.

Induction of endothelial NO synthase by hydrogen peroxide via a Ca(2+)/calmodulin-dependent protein kinase II/janus kinase 2-dependent pathway.

We have recently demonstrated that hydrogen peroxide (H(2)O(2)) is an extremely potent stimulus of endothelial NO synthase (eNOS) gene expression. The present study was designed to identify the signaling mechanisms mediating this response. Induction of eNOS expression by H(2)O(2) was found to be Ca(2+) dependent, inasmuch as it was blocked by BAPTA-AM. Further studies have indicated that Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) plays a critical role in mediating this response. Immunocytochemical staining with an anti-CaM kinase II antibody confirmed the expression of CaM kinase II in cultured bovine aortic endothelial cells. H(2)O(2) induced autophosphorylation of CaM kinase II and increased the activity of the enzyme, as assessed by an in-gel kinase assay. A specific inhibitor for CaM kinase II, KN93, and a calmodulin antagonist, W-7, attenuated eNOS induction by H(2)O(2). Further studies have indicated that janus kinase 2 is important in mediating increased eNOS expression in response to H(2)O(2) and likely is downstream from CaM kinase II. In conclusion, these data provide the first evidence that CaM kinase II plays a critical role in endothelial redox signaling. Regulation of eNOS via this pathway may represent an important vascular adaptation to oxidant stress.

The histone deacetylase inhibitor MS-275 induces caspase-dependent apoptosis in B-cell chronic lymphocytic leukemia cells.

MS-275 is a histone deacetylase (HDAC) inhibitor that has been reported to mediate its cytotoxic effect through generation of reactive oxygen species (ROS) in proliferating hematopoietic cell lines. We examined efficacy of MS-275 in nonproliferating chronic lymphocytic leukemia (CLL) cells from patients. In these cells, MS-275 demonstrated an in vitro LC(50) that was one log lower than for normal mononuclear cells. Following MS-275 treatment, histones H3 and H4 showed increased acetylation and HDAC enzymatic activity was reduced. Caspase-8, -9, and -3 were activated, and caspase substrates PARP and BID were cleaved. Additionally, FLICE-inhibitory protein (FLIP) was downmodulated following MS-275 incubation. MS-275 treatment caused detectable ROS generation after 15 h of incubation, which was blocked by the caspase inhibitor Z-VAD-fmk. Overexpression of Bcl-2 protein protected against MS-275-induced apoptosis. These data demonstrate that MS-275 is a promising therapy for the treatment of CLL, but that in contrast to previous reports, ROS generation does not precede commitment to apoptosis. Similar to many other therapeutic targets, MS-275-mediated apoptosis is reduced by overexpression of Bcl-2, justifying strategies to combine HDAC inhibitors with Bcl-2 antagonists.

Differential Expression of Cell Cycle Regulators During Hyperplastic and Hypertrophic Growth of Broiler Subcutaneous Adipose Tissue.

Hyperplastic growth and hypertrophic growth within adipose tissue is tightly associated with cell cycle activity. In this study, CCNG2 and CDKN2C were found to be correlated with cell cycle inhibition during fat cell differentiation, whereas CCND3, CCNA1, and ANAPC5 were positively associated with cell cycle activity during fat cell proliferation after selection based on GEO datasets available on the NCBI website. The findings were validated through comparison of expressions of these genes among different tissues/fractions in broiler chickens and time points during primary cell culture using quantitative real-time PCR. Development of broiler subcutaneous adipose tissue was investigated on embryonic days 15 and 17 and on post-hatch days 0, 5, 11, and 33 using H&E staining and PCNA immunostaining with DAPI counter stain. In addition, mRNA expressions of five cell cycle regulators as well as precursor cell and adipocyte markers were measured at those time points. The results suggest that cellular proliferation activity decreased as the fat pad grows, but a population of precursor cells seemed to be maintained until post-hatch day 5 despite increasing differentiation activity. Hypertrophic growth gradually intensified despite a slight cessation on post-hatch day 0 due to increased energy expenditure during hatching and delayed food access. From post-hatch day 5 to day 11, most of the precursor cells may become differentiated. After post-hatch day 11, hyperplastic growth seemed to slow, while hypertrophic growth may become dominant. This study provides further understanding about broiler fat tissue development which is imperative for effective control of fat deposition.

Location of Ge and extra-framework species in the zeolite ITQ-24.

The germanosilicate ITQ-24 (IWR framework type) was synthesized in fluoride medium using 1,3,5-tris(1,2-dimethylimidazolium) benzene as the structure directing agent (SDA). A structure analysis of the as-synthesized ITQ-24 material using synchrotron powder diffraction data and difference electron density calculations have allowed the fluoride ions and the germanium atoms to be located and the conformation of the SDA to be determined. The benzyl ring is perpendicular to the b axis with the three imidazolium moieties forming a "T-shaped" arrangement. Ge atoms replace some of the Si in the double-4-ring (d4r) and in one of the single-4-rings (s4r). The other s4r contains only Si. Fluoride ions are in the d4r units. Initially, the space group Cmmm (highest possible symmetry) was assumed, but the framework geometry was strained. An independent evaluation of the symmetry using the powder charge flipping algorithm in Superflip led to a successful refinement with reasonable geometry and a refined composition of |[(C6H3)(C7H10N2)3]2F2|[Si(40.2)Ge(15.8)O112] in the space group Pban.

Self-assembling nucleic acid delivery vehicles via linear, water-soluble, cyclodextrin-containing polymers.

Non-viral (synthetic) nucleic acid delivery systems have the potential to provide for the practical application of nucleic acid-based therapeutics. We have designed and prepared a tunable, non-viral nucleic acid delivery system that self-assembles with nucleic acids and centers around a new class of polymeric materials; namely, linear, water-soluble cyclodextrin-containing polymers. The relationships between polymer structure and gene delivery are illustrated, and the roles of the cyclodextrin moieties for minimizing toxicity and forming inclusion complexes in the self-assembly processes are highlighted. This vehicle is the first example of a polymer-based gene delivery system formed entirely by self-assembly.

Transplacental uptake of WR 2721 by the rat embryo.

On day 14 post-conception, near the end of the period of major organogenesis, pregnant rats were injected intravenously or intraperitoneally with WR 2721 spiked with 14C-WR 2721. The radioprotectant was shown to cross the placenta rapidly when administered by either route, and the concentration of WR 2721 in the embryos, placentae, and maternal blood plasma was determined during the period 5 to 90 minutes following administration. The concentration of WR 2721 increased continuously in the embryos during this period and did so against a decreasing concentration in the maternal blood. Injection of WR 2721 at 100 mg/kg of maternal body weight resulted in the presence of 8-9-mg/kg embryo weight; this embryo level is about 1/2 the injected dose of WR 2721 currently being used in human radiotherapy trials, that is, 20 mg/kg (740 mg/m2) body weight. Previous toxicity studies of 9, 11, and 14 day rat embryos have shown that this 100 mg/kg dose is much below the level which produces embryotoxic effects.

Circumvention of the tumor membrane barrier to WR-2721 absorption by reduction of drug hydrophilicity.

In attempting to account for the ability of most solid tumors to restrict the absorption of WR-2721, aminopropyl-aminoethylphosphorothioate, we examined a number of drug characteristics which might allow for this restriction, and observed that drug hydrophilicity was a major contributing factor. When the highly hydrophilic WR-2721 was dephosphorylated, the drug became less hydrophilic and could readily cross tumor cell membranes. In addition, conventional radioprotectants, such as cysteine and mercaptoethylamine, were shown to be less hydrophilic than WR-2721 and also to cross tumor membranes readily. Therefore, drug hydrophilicity would appear to be the factor underlying the ability of WR-2721 to selectively protect normal tissues while most other protectors alter the radiation resistance of normal and tumor tissue alike. A red blood cell model for studying this problem in greater detail is described.