Immunoproliferative small intestinal disease: prolonged 30-year course without development of lymphoma.

Immunoproliferative small intestinal disease (IPSID) is mostly found in young adults of low socioeconomic class in developing countries. This condition is characterized by a dense lymphoplasmacytic infiltrate beneath the epithelium in the duodenal and proximal jejunal mucosa and in the mesenteric lymph nodes. In two thirds of cases, the involved lymphocytes elaborate an anomalous alpha-heavy chain protein. The etiology of this disease is unclear, although various parasitic, genetic, and toxic mechanisms have been proposed. Half of all IPSID patients will be found at diagnosis to have a concurrent intestinal B-cell lymphoma, and most of the remaining patients develop frank lymphoma within a few years. Although most reports of IPSID are from developing nations or indigent immigrant populations within Western countries, four cases of an IPSID-like condition have been documented in white women. Furthermore, although many IPSID patients progress to high grade indeterminate-type lymphoma within a few years of initial presentation, there have been occasional reports of long term survival without lymphomatous conversion. Here, we present an atypical case of IPSID--a California native who, though of Mexican heritage, had resided in the United States his entire life and did not belong to an indigent population. This patient had biopsy-proven IPSID that progressed over 30 yr but never exhibited lymphomatous conversion despite end stage intestinal stasis and recurrent obstruction, culminating in death. Our case calls into question some current assumptions about the prelymphomatous nature of this disease.

Sucrase-alpha-dextrinase in the spontaneously diabetic BioBreed Wistar rat: altered intracellular carbohydrate processing.

Sucrase-alpha-dextrinase (S-D), a glycoprotein hydrolase in the border surface of the enterocyte, is altered in congenitally diabetic BioBreed Wistar (BB(d)) rats. Its intracellular assembly was examined in the current studies. Following pulse-chase experiments, S--D was specifically immuno-isolated from ER-Golgi and brush border membranes, and examined by SDS-PAGE and autoradiography. While synthesis and co-translational glycosylation of an immature species, P(i), in the ER proceeded normally, post-translational maturation of the N-linked carbohydrate chains was altered in the BB(d) rat. The mature species, P(m), was 10 kDa larger in BB(d) than in normal rats, and approximately 25% of its N-linked chains remained immature. The difference in mass was attributed to an appreciably larger mass of the O-linked chains of P(m) in BB(d) rats. In vivo kinetics of intracellular assembly displayed a delay in the appearance of P(m) in Golgi (Wistar, 15 min; BB(d), 30--60 min). These experiments, revealing structural alterations in congenital diabetes suggest an important role for intracellular glycosylation in the orderly assembly and processing of brush border S-D in the enterocyte.

Intestinal aminooligopeptidase in diabetic BioBreed rat: altered posttranslational processing and trafficking.

The structure of aminooligopeptidase (AOP), an intestinal brush-border digestive hydrolase, is abnormal in human diabetes and in the congenitally diabetic BioBreed Wistar (BB(d)) rat. Its assembly in the BB(d) rat was examined. After normal initial synthesis and assembly of immature AOP precursor (AOP(i)) with high-mannose N-linked chains in the endoplasmic reticulum (ER), processing of N-linked glycans in Golgi yielded a smaller than normal mature AOP precursor (AOP(m)) with persistence of some high-mannose N-linked chains. Deglycosylation analyses suggested that the mass difference could be attributed to a lower mass of N-linked with unaltered O-linked glycans in AOP(m) of the diabetic rat. Intrajejunal pulse-chase experiments revealed that the conversion of AOP(i) to AOP(m) occurred at 30 min of chase in normal rats but at 60-90 min in diabetic rats, reflecting delay in ER-to-Golgi transport or a slower processing of high-mannose chains. Once maximal transport to Golgi was achieved, the residence time in Golgi was shortened in diabetes. This altered processing of the precursor accounted for the altered structure of AOP in diabetes.

Risk/risk trade-offs in pesticide regulation: an exploratory analysis of the public health effects of a ban on organophosphate and carbamate pesticides.

Efforts to reduce pesticide-related risks to consumers and farmworkers often neglect the possibility that measures to reduce the target risk may introduce or enhance countervailing risks. These may arise from substitute pesticides or pest-control practices, from increased levels of pests or pest-related hazards, from increased levels of toxic natural pesticides in plants, from increased costs and decreased consumption of health-enhancing fruits and vegetables, or from direct income effects on consumers and farmers. The effect of the countervailing risks may partially or completely offset the reduction in the target risk. A risk-trade-off analysis was conducted of a potential ban on the use of organophosphate and carbamate (OP/Carbamate) insecticides in U.S. agriculture. Although this scenario is extreme, it has the analytic virtue of dispensing with the infinite number of "next-best" OP/Carbamates that might be substituted for specific combinations of crops and pests should only selected uses be banned. The analysis relies on detailed descriptions of the alternative pesticides and pest-control measures that would be used for each of 14 major crops. The effects of pest-control cost changes on prices and consumption and effects on consumer and producer incomes are projected using a general-equilibrium economic model. Several countervailing risks that may be significant were found, including acute toxicity to farmworkers from substitute pesticides, cancer and noncancer risks from substitute pesticides, and mortality induced by changes in disposable income. Other countervailing risks are more difficult to estimate or weigh. Potential increases in natural plant pesticides following an OP/Carbamate ban are discussed but data are lacking to quantify the effects. Changes in diet following the ban have both positive and negative effects, and the ultimate change is difficult to estimate. Although a net risk cannot be estimated, several approaches were illustrated that would be useful in risk-trade-off analyses. Key factors complicating comprehensive analysis of risk/risk trade-offs for pesticides were also identified, including data gaps and shortcomings of current risk assessment methods.

Correlations among tumor types in mouse cancer bioassays: liver adenomas, liver carcinomas, leukemias and lymphomas.

In an examination of rodent bioassays Young and Gries [Young S.S., and Gries C.L. Exploration of the negative correlation between proliferative hepatocellular lesions and lymphoma in rats and mice--establishment and implications. Fundam. Appl. Toxicol. 1984: 4: 632-6401 and Haseman et al. [Haseman J.K., et al. Body weight-tumor incidence correlations in long-term rodent carcinogenicity studies. Toxicol. Pathol. 1997: 25: 256-263] noticed that there is a negative correlation (anticorrelation) between development of liver tumors and leukemia or lymphomas. If an animal has a lymphoma or leukemia it is less likely to develop liver tumors. These studies noted that this applies to several strains of animals. The anticorrelation appeared in control animals. In this paper we study this anticorrelation in the quarter of a million rodents exposed in the Carcinogenesis Bioassay Database System (CBDS) database of the National Toxicology program in both control and dosed animals. We failed to completely replicate Young and Gries or Haseman et al. However, when benign liver tumors (adenomas or nodules) and malignant liver tumors (carcinomas) are considered separately AND different leukemia and lymphoma types are considered, a strong anticorrelation appears. We identify survival and the time period (in years) during which the bioassay was completed as an important factor in interpreting correlations and anticorrelations. Differences between liver adenomas and carcinomas and lymphoma types contribute to a more general question of grouping of the individual tumor types. In our classification scheme (originally developed about 1986 by Dr. Bailar) an effort is made to distinguish benign and malignant neoplasms, while other investigators group all tumors at a specific site. For many analyses liver adenomas and carcinomas have been lumped together. This is because it is suspected that the diagnoses by pathologists may not distinguish, or ignore the (benign) adenoma when a (malignant) carcinoma is present. Possible biological differences in tumor mechanisms may require separate evaluation of these tumors with all the dangers of "pathologist bias" that this introduces.

Anticarcinogenic responses in rodent cancer bioassays are not explained by random effects.

Anticarcinogenicity in a long-term rodent bioassay is defined as a statistically significant decrease of a specific tumor type in a dosed group following chemical exposure. About 92% of chemicals tested by the National Toxicology Program prior to 1983 reveal at least one site with a significant (p < or = 0.05) tumor rate decrease in one or more tested groups, a result consistent with those of J. K. Haseman and F. M. Johnson (1996, Mutat. Res. 350, 131-141) for a database of recently tested chemicals. Detection of tumor decreases in a specific site can be explained not only by biological effects, but also as a result of random variability in the background tumor rates, decreases in body weight, or decreases in survival of treated animals. This paper evaluates the rate of false-positive anticarcinogenic findings due to random effects (variations in tumor rates and the multiple comparisons undertaken in evaluating a bioassay), while a companion paper addresses the influence of weight and survival depression. Monte-Carlo simulation was conducted to assess the contribution of random effects. This contribution was found to be important even when a statistical significance cutoff of p0 < or = 0.05 was chosen. If a more stringent statistical criterion was used (p0 < or = 0.01 or p < or = 0.005), the proportion of false positive determinations diminishes. The number of anticarcinogens in the database remains substantially higher than predicted by the stimulations. An examination of the distribution of all p values (T. Schweder and E. Spjøtvoll, 1982, Biometrika 69, 493-502) also indicates that statistically significant anticarcinogenic responses are found in the database at a higher rate than would result from purely random responses. Finally, the cross-species prediction of anticarcinogenic responses was examined in a manner similar to a study of cross-species prediction of carcinogenic responses (G. M. Gray et al., 1995, Reg. Toxicol. Pharmacol. 20, 281-301). The analyses show that anticarcinogenic effects in one rodent species predict well anticarcinogenic effect in another rodent species. It seems likely that biological factors are involved in anticarcinogenic responses observed in rodent cancer bioassays.

Weight and survival depression in rodent bioassays with and without tumor decreases.

It has been suggested that the decreased tumor rates (anticarcinogenicity) commonly observed in the National Toxicology Program (NTP) rodent bioassays may be caused by compound-induced decreases in body weight or decreases in survival of treated animals. In this study, weight decrement and survival depression following chemical treatment was studied for those chemicals which induce site specific decreases in tumor rates (anticarcinogens) and those which do not (non-anticarcinogens) in a database of 312 chemicals tested in the NTP bioassay program prior to 1983. There is an evident difference in weight depression and animal survival between anticarcinogens and non-anticarcinogens but it is small relative to the variability between chemicals in the two groups. We argue that weight or survival depression cannot, in a simple way, explain the difference between anticarcinogenic and non-anticarcinogenic chemicals. In fact, there are a number of chemicals that are anticarcinogenic with no evidence of weight or survival depression and many chemicals that cause significant weight or survival decreases with no apparent anticarcinogenic effects. There is a small but statistically insignificant relationship between the degree of weight depression and the number of tumor sites found to have lower tumor rates in treated animals. These analyses suggest that biological factors other than weight and survival depression are involved in decreased tumor rates in rodent bioassays. These results, and those of the companion paper (I. Linkov et al., this issue), suggest that the anticarcinogenic responses observed in rodent cancer bioassays should be carefully considered in evaluations of the overall carcinogenic potential of chemicals.

An empirical examination of factors influencing prediction of carcinogenic hazard across species.

This study was stimulated by a recent U.S. Environmental Protection Agency (EPA, 1994) statement in draft environmental carcinogen risk assessment guidelines: "Several kinds of observations from animal studies can contribute to the judgment whether animal responses indicate a significant carcinogenic hazard to humans." We have investigated each of these kinds of observation using the cancer bioassay data system database. We obtained concordances from rat to mouse (and vice versa) for various subgroups of chemicals as follows: chemicals that induced tumors at multiple sites, chemicals that induce cancer in both sexes, chemicals that display reduced latency, and chemicals increasing the rates of rare tumors. The concordances are much higher for these chemical subgroups than the chemical groups that induce tumor at a single site, in only one sex, or without reduced latency, respectively. Thus, our findings support some of the EPA's suggested factors.

The ability of predicted internal dose measures to reconcile tumor bioassay data for chloroform.

PBPK models are developed in the hope that they will improve our ability to extrapolate from one species to another and from one exposure regime to another. Evidence that a dose measure was successful at reconciling the available animal bioassay data would be encouraging. It would give us some confidence that the dose measure (as evaluated by a PBPK model) might yield reasonable predictions for yet other species (e.g., humans) and other dose routes. We have investigated the ability of a modified version of the Corley et al. (R. A. Corley, A. L. Mendrala, F. A. Smith, D. A. Staats, M. L. Gargas, R. B. Conolly, M. E. Andersen, and R. H. Reitz, 1990, Toxicol. Appl. Pharmacol. 103, 512-527) PBPK model for chloroform to reconcile the available bioassay data. Two rate-dependent dose measures, maximal rate of metabolism in the liver, and percentage of hepatocytes killed per day performed well at reconciling the rodent liver bioassay data, while all rate-independent dose measures performed less well. In contrast, none of the PBPK dose measures were capable of reconciling the rat and mouse kidney tumor response data. Here, administered dose scaled to body surface area performed the best.

Conformationally defined cyclohexyl carnitine analogs.

Three diastereoisomers of racemic (3-carboxy-2-hydroxy-1-cyclohexyl)trimethylammonium chloride [C10H20NO3+.Cl-; (1S,2S,3S) (2), (1R,2S,3S) (3) and (1S,2R,3S) (4)] were designed as rigid analogs for different low-energy conformational states of carnitine [(1), (3-carboxy-2-hydroxy-1-propyl)trimethylammonium chloride]. Structures (2)-(4) all assume a chair conformation in the solid state, in which the bulky trimethylammonio group occupies the equatorial position. As such, the orientations about C2-C3 in (2), (3) and (4) are all essentially the same as that found for (1) in the solid state (torsion angles for C1-C2-C3-N1 near 180 degrees), while the orientations about C1-C2 in (2)-(4) are such that each diastereoisomer contains a different one of the three possible low-energy staggered conformations predicted for (1) in solution. Comparisons between (1) and (2)-(4) in the solid state revealed that diastereoisomers (2), (3) and (4) provide rigid models for the major low-energy conformations of carnitine.

Use of probabilistic expert judgment in uncertainty analysis of carcinogenic potency.

A new approach to characterizing the state of knowledge about carcinogenic potency is described. In this approach, the carcinogenic risk posed by a specific dose is characterized by a probability distribution, indicating the relative likelihood of different risk estimates. The approach utilizes expert judgment and a probability tree and is illustrated in a case study of chloroform exposure. Experts in cancer biology/toxicology, pharmacokinetics, and dose-response modeling were identified by a panel of science-policy specialists. In a workshop, experts reviewed the chloroform data, received training in probability elicitation, and constructed a consensual probability tree based on biological theories of cancer causation. Distributions of carcinogenic risk were developed based on the probability tree, chloroform data, judgmental probabilities provided by the experts, and classical statistical techniques. Risk distributions varied considerably between experts, with some predicting essentially no risk from 100 ppb chloroform in drinking water while other have at least some probability on risks generally considered of regulatory significance. Estimated human risk was much lower when extrapolating from liver tumors in animals than from kidney tumors. Issues of scientific disagreement leading to different risk distributions between experts are discussed. The resulting risk distributions are compared to standard EPA risk calculations for the same exposure scenario as well as to the expert judgement of epidemiologists about cancer risks of chlorinated drinking water. Issues in combining expert judgments are discussed, and several alternative methods are presented. Strengths and weaknesses of the distributional approach are discussed.

A distributional approach to characterizing low-dose cancer risk.

Since cancer risk at very low doses cannot be directly measured in humans or animals, mathematical extrapolation models and scientific judgment are required. This article demonstrates a probabilistic approach to carcinogen risk assessment that employs probability trees, subjective probabilities, and standard bootstrapping procedures. The probabilistic approach is applied to the carcinogenic risk of formaldehyde in environmental and occupational settings. Sensitivity analyses illustrate conditional estimates of risk for each path in the probability tree. Fundamental mechanistic uncertainties are characterized. A strength of the analysis is the explicit treatment of alternative beliefs about pharmacokinetics and pharmacodynamics. The resulting probability distributions on cancer risk are compared with the point estimates reported by federal agencies. Limitations of the approach are discussed as well as future research directions.

Nonlinear-optical studies of molybdenum metal organics.

We have measured the third-order susceptibility, X((3)), versus concentration for a number of molybdenum-based metal-organic complexes in solution, using independent degenerate four-wave mixing and Z-scan techniques. Good agreement was obtained between the degenerate four-wave mixing and Z-scan measurements. The variation of X((3)) with concentration yielded the second-order hyperpolarizability gamma. A close correlation was observed between the number of delocalized pi electrons and the magnitude of gamma.

Intestinal lactase: maturational excess expression of mRNA over enzyme protein.

The mechanism of decline of intestinal lactase during mammalian development remains uncertain. Despite a major loss of catalytic activity, lactase mRNA appears to persist at detectable concentrations in adult rats. We quantified lactase activity, total lactase protein, and lactase mRNA in rats aged 7, 11, 15, 18, 22, 30, and 60 days using the 7S ribosomal RNA as the developmental control. The active lactase fraction was 0.81 of total lactase for all age groups except 60-day-old animals, in which it declined to 0.60 (P = 0.004), indicating that conversion of active lactase to inactive species contributed to the lower activity in the adult. Northern blots revealed a single discrete 6.8-kb message at all ages. Although lactase activity and immunoprotein decreased coordinately to a minimum by day 30 (20% of the 7-day value), lactase mRNA doubled to a maximum at day 22 and was maintained at 7-day concentrations even in 60-day adults. The lactase mRNA-to-protein ratio was low at 7 days (0.19) but more than doubled (0.50) by 22 days, achieved a fivefold increase (1.0) by 30 days, and persisted at 0.77 in adults. The relative excess of lactase message during maturation suggests that translational or post-translational events may be paramount in the developmental regulation of lactase gene expression.

The challenge of risk characterization: current practice and future directions.

Risk characterization is perhaps the most important part of risk assessment. As currently practiced, risk characterizations do not convey the degree of uncertainty in a risk estimate to risk managers, Congress, the press, and the public. Here, we use a framework put forth by an ad hoc study group of industry and government scientists and academics to critique the risk characterizations contained in two risks assessments of gasoline vapor. After discussing the strengths and weaknesses of each assessment's risk characterization, we detail an alternative approach that conveys estimates in the form of a probability distribution. The distributional approach can make use of all relevant scientific data and knowledge, including alternative data sets and all plausible mechanistic theories of carcinogenesis. As a result, this approach facilitates better public health decisions than current risk characterization procedures. We discuss methodological issues, as well as strengths and weaknesses of the distributional approach.

Sucrase-alpha-dextrinase in the rat. Postinsertional conversion to inactive molecular species by a carbohydrate-free diet.

Absence of dietary carbohydrate decreases both activities of intestinal brush border sucrase-alpha-dextrinase. We examined the molecular mechanism causing this decrease. Adult rats were fed chow (70% CHO) or matched carbohydrate-free (CHO-free) diet for 7 d. Sucrase activity decreased by 50% in whole homogenates and brush borders. Enzyme kinetics revealed no change in sucrose affinity (CHO-free Km = 18 mM, chow Km = 21 mM), but fewer active sites (CHO-free Vmax = 2,720, chow Vmax = 5,000 mumol/min per g protein). Intraintestinal pulse-labeling of [35S]methionine in vivo revealed no differences in incorporation into sucrase. Immunoreactive sucrase protein, assayed by ELISA and rocket immunoelectrophoresis, increased twofold per milliunit of sucrase enzymatic activity in CHO-free jejunum. Total immunosucrase (St), the sum of active and inactive enzyme (St = Sa+Si), was unchanged with carbohydrate withdrawal, but > 50% of the sucrase protein became inactive. SDS-PAGE of sucrase immunoprecipitates revealed alteration of alpha, beta, and gamma subunits in CHO-free animals: (a) alpha and beta subunits migrated farther (mass change--2 kD); and (b) the alpha subunit became diffuse or was a doublet and was less abundant than the beta subunit. Rather than representing loss of sucrase protein, the decline in sucrase activity is achieved with structural subunit changes, probably involving postinsertional processing.