Problems in determination of skeletal lead burden in archaeological samples: an example from the First African Baptist Church population.

Human bone lead content has been demonstrated to be related to socioeconomic status, occupation and other social and environmental correlates. Skeletal tissue samples from 135 individuals from an early nineteenth century Philadelphia cemetery (First African Baptist Church) were studied by electrothermal atomic absorption spectrometry and X-ray fluorescence for lead content. High bone lead levels led to investigation of possible diagenetic effects. These were investigated by several different approaches including distribution of lead within bone by X-ray fluorescence, histological preservation, soil lead concentration and acidity as well as location and depth of burial. Bone lead levels were very high in children, exceeding those of the adult population that were buried in the cemetery, and also those of present day adults. The antemortem age-related increase in bone lead, reported in other studies, was not evidenced in this population. Lead was evenly deposited in areas of taphonomic bone destruction. Synchrotron X-ray fluorescence studies revealed no consistent pattern of lead microdistribution within the bone. Our conclusions are that postmortem diagenesis of lead ion has penetrated these archaeological bones to a degree that makes their original bone lead content irretrievable by any known method. Increased bone porosity is most likely responsible for the very high levels of lead found in bones of newborns and children.

Lead in bone. I. Direct analysis for lead in milligram quantities of bone ash by graphite furnace atomic absorption spectroscopy.

A method for direct lead content analysis of milligram quantities of bone ash by flameless atomic absorption spectroscopy is described. Bone ash (25 mg) is dissolved with HNO3 and diluted with H2O and La2O3 (1,000 micrograms/ml) solution. Lanthanum ion is used to suppress matrix interferences possibly arising in part from sulfate components of the bone ash. Two bulk bone samples (about 14 and 60 micrograms Pb/g ash, respectively) were used to determine daily, within-day, and overall variability of the method. Values for "low lead" bone samples were 14.08 +/- 1.74 (SD) microgram Pb/g ash and for "high lead" bone samples were 60.85 +/- 5.24 (SD) microgram Pb/b ash. The overall value of 58 lead recovery determinations from bone ash analysis was 103.5% (+/- 12.9% SD). These values compare favorably with results previously reported using gram amounts of sample.

Effect of adrenalectomy on the metabolism of glucose in obese (C57 Bl/6J ob/ob) mice.

The genetically obese mouse, C57 B1/6J ob/ob, has been suggested as an appropriate model for the study of obesity associated with diabetes mellitus. Employing glucose 14C(microliter) as a tracer, the data presented here indicate that obese mice are able to clear glucose from the blood compartment at the same rate as their lean littermates. This was demonstrated with or without an associated cold glucose load. The abnormal glucose tolerance curves observed in the obese animals may be a result of secretion of glucose into the blood. Removal of the adrenal glands from the obese mice and their lean littermate does not impair their ability to clear a glucose load from the vascular compartment. The capacity for endogenous glucose secretion of ob/ob mice is severely curtailed by adrenalectomy, in that the glucose tolerance curves of these adrenalectomized animals become similar to those of sham-operated lean littermates. Thus, it appears that a considerable component of the hyperglycemia in ob/ob mice reflects major adrenal involvement that is activated by stress, ie, ether anesthesia and blood sampling. The hyperglycemia in ob/ob mice may reflect glucocorticoid-dependent gluconeogenesis.

Gastric mucosal damage due to aspirin and copper aspirinate assessed by gastric mucosal potential difference changes.

When the gastric mucosa is damaged by antiinflammatory agents, such as aspirin, gastric mucosal potential difference (GPD) decreases and may or may not return to predamage values after the agent is removed. The magnitude and time course of the gastric potential difference changes have been suggested as a measure of mucosal damage. Male Sprague-Dawley rats were fasted, anesthetized, and surgically prepared for measurement of GPD by placement of electrodes in the gastric lumen and spleen. Test mixtures of aspirin, copper aspirinate, copper sulfate, or mixtures of aspirin and copper sulfate were administered by gavage, and subsequent changes in GPD were recorded. The area between the extrapolated control (baseline) GPD and the damage GPD was determined. The product of this area and the maximum change in GPD, the Reizindex (RI), was calculated. Values for all copper (II)-containing systems, including those with no aspirin, were significantly greater than aspirin alone. It was concluded that factors other than mucosal damage may contribute to a reduction in GPD and, therefore, an increase in Reizindex.

Ethanol-induced hypothermia and hyperglycemia in genetically obese mice.

Blood glucose and rectal temperatures were monitored in two strains of genetically obese mice (C57 BL/6J ob/ob) prior to and following intragastric ethanol administration in an attempt to relate the hypothermic response to ethanol to extracellular glucose concentration. In contrast to expectation, ethanol administration was typically associated with a hyperglycemia and a hypothermic response. In the ob/ob genotype, the hypothermic response was associated with pronounced hyperglycemia which was more emphatic in older animals. The data support the conclusion that ethanol-induced hypothermia is independent of blood glucose levels. In light of the known sensitivity of ob/ob mice to insulin, it is suggested further that the observed hypothermic response was not a function of the animals' ability to transport glucose into peripheral cells. The observed hyperglycemia of the obese animals was most likely stress-related.

Selected aspects of the spatial distribution of lead in bone.

The macrodistribution of lead within the skeletal system is not uniform, and is clearly dependent on bone type (compact or trabecular), age and (to a lesser extent) gender. During active growth, lead deposition favors trabecular bones sites but in adulthood lead accumulates more actively in compact bone. The extent and patterns of the higher variability in trabecular bone sites require better definition through further study. Lead tends to be deposited in bone at sites of most active calcification at the time it is absorbed. During the first 15 years of life the bone growth centers dominate the process of calcification. In adulthood osteon remodelling and trabecular plate mineral exchanges are the principal calcifying sites. Both patterns probably deteriorate in old age. Microdeposition patterns of lead could be expected to conform to these changes. However, it is difficult to integrate the reported information into a clearly defined chronological sequence, because many of the studies did not control for age and other variables with sufficient precision. Furthermore, kinetic studies suggest lead is deposited in multiple sites not only among soft tissues but also within bone in molecular forms possessing varying affinities. Current efforts to design instrumentation for in vivo diagnostic lead measurements as well as therapeutic methods to mobilize and remove lead in bone require more precise knowledge than is now available concerning lead macrodistribution, microdistribution and anatomic sites of kinetic compartments. Appropriately designed studies employing contemporary, sensitive techniques involving isotopes, microradiographs, autoradiographs, histology, tomography, and high energy x-ray fluorescence (synchrotron radiation-induced x-ray emission-SRIXE) are now capable of generating the needed data.

Comparison of two in vitro methods of bone lead analysis and the implications for in vivo measurements.

Atomic absorption spectrometry and x-ray fluorescence have been used to determine the lead content of metatarsal and tibia bone samples. For a range of bone lead levels from 6.5 to 83 micrograms g-1 of ashed bone there is no evidence of a systematic difference between the two techniques of more than 1 microgram g-1. There is, however, some evidence that random differences between the two in vitro analyses applied to the same bone sample are larger than can be accounted for by known measurement uncertainties. Variations in bone composition could account for these differences. Because the x-ray fluorescence technique is applied in an identical way to in vivo analysis, it is concluded that the uncertainties in in vivo measurements are small.

Hydrolysis of tetrakis-mu-acetylsalicylato-dicopper(II).

Hydrolysis rates of acetylsalicylate in the free acid and anion forms and in the dilute solution of the copper complex, tetrakis-mu-acetylsalicylato-dicopper(II), were compared. The hydrolysis rate was unaffected by the presence of copper(II). The pH-dependent rate was 3.64 x 10(-4) mmole/liter/min per pH unit at 37 degrees in Ringer's solution. The synthetic procedures commonly used for the preparation of this compound yield a product contaminated with salicylate. This contamination is avoided by synthesis of the compound in methanol, which yields a complex containing no measurable salicylate.

Comparison of aspirin and copper aspirinate with respect to gastric mucosal damage in the rat.

The copper salt of aspirin has been compared with aspirin in terms of damage to mucosal tissue. Using a protein-bound dye to highlight erosions, it has been found that copper aspirinate is at least as damaging as aspirin itself. This finding is not in agreement with previously published claims. Copper aspirinate produced more widespread superficial erosions and slightly less deep erosions than aspirin alone. Mixtures of the copper(II) ion and aspirin produced results similar to copper aspirinate, suggesting that the hydrolysis products of copper aspirinate, copper(II) ion and aspirin, together may be especially damaging to the mucosa. Copper alone was not damaging, but aspirin alone yielded intermediate results. Short incubation times produced only erosions (no ulcers), which were clearly differentiated into two classes by depth of color. Histological examination verified this classification into superficial and deep erosions.

Core temperature response to immersed bicycle ergometer exercise at water temperatures of 21 degrees, 25 degrees, and 29 degrees C.

A bicycle ergometer modified for aquatic exercise was used to determine the effects of immersion on core temperature during submaximal exercise at different water temperatures. An exercise intensity (60% of maximal oxygen consumption) and duration (30 minutes) considered appropriate for cardiovascular conditioning were used. These data will be useful in cardiovascular and leg-strengthening hydrotherapy programs. Rectal temperature, skin temperature, and a rating of thermal comfort were studied in five normal men (14.8% +/- 5.6% fat) during headout immersion at water temperatures of 21.1 degrees, 25.3 degrees, and 29.4 degrees C and exercise in air of 21.1 degrees C. Subjects were immersed for 30 minutes during static and exercise (63% +/- 0.6% maximal oxygen consumption) conditions. Data were collected every 5 minutes and analyzed by repeated measured analysis of variance. At water temperatures, rectal temperature fell from control during static immersion (p less than or equal to 0.05) and was lower than control at the end of the 30-minute recovery period (p less than or equal to 0.05). During exercise there was no change in rectal temperature at water temperatures of 21.1 degrees and 25.3 degrees C; however, rectal temperature rose at water temperatures of 29.4 degrees (p less than or equal to 0.05) and air 21.1 degrees C (p less than or equal to 0.05). At the end of recovery rectal temperature was lower than control at water temperatures 21.1 degrees C (p less than or equal to 0.05) and greater than control at water temperatures 29.4 degrees C (p less than or equal to 0.05). There was no change from control in rectal temperatures at water temperatures 25.3 degrees C and air at 21.1 degrees C. These results indicate that immersion in 25.3 degrees and 21.1 degrees C water effectively attenuates the rise in rectal temperature during exercise at 63% of maximal oxygen consumption, whereas immersion in 29.4 degrees C water does not. In addition, both skin and rectal temperatures affect perception of thermal state but do not give the subjects an accurate estimation of thermal balance.

Suppression of shivering by breath holding, relaxation, mental arithmetic, and warm water ingestion.

Four methods of suppressing cold-induced shivering were evaluated in 26 young male volunteers exposed to 0.0 +/- 1.0 degrees C air for 135 min. Voluntary relaxation of musculature (R), breath holding (BH), warm (50 degrees C) water ingestion (W), and performance of a mental arithmetic task (MA) were applied in a counterbalanced order following 2 h of cold exposure. Surface electromyograms of seven muscles were recorded and converted to root mean square voltage (RMS) as a measure of shivering intensity. Mean skin and rectal temperatures decreased significantly, 4.9 degrees C and 0.3 degrees C, respectively (p < or = 0.01). Mean reduction of EMG activity was 35% during R, 24% during BH, 18% during MA, and 5% during W. R was significantly more effective than BH, MA, and W, and BH and MA were significantly more effective than W in reducing shivering. These results indicate that, at small decreases in rectal temperature, shivering can be voluntarily suppressed to some extent during relaxation, breath holding, and mental arithmetic.

Lead in bone. IV. Distribution of lead in the human skeleton.

Flameless atomic absorption spectroscopy was used to measure lead concentrations in samples from 5 selected human skeletal sites (tibia, skull, rib, ilium, and vertebra) obtained from 134 hospital autopsies. Lead was distributed unequally among the different bones in distinct patterns that were age-, and to some extent, sex-dependent. To estimate lead concentration of the entire skeleton, all skeletal bones were divided into 5 groups based on their approximate compact/trabecular bone ratios, considering each of our 5 sampled sites to be the prototype for each such group. Regression analysis of the 10 possible bone site pair values at different ages yielded age-related constants. These constants were incorporated into an equation we developed that can be used both to estimate mean skeletal lead concentration (Pb) of the entire body skeleton and also to predict the lead concentration at any of the other 4 bone sites if any 1 of the 5 is measured. Applications of these data to in vivo bone lead measurements are detailed with respect to selection of the site to be measured, estimation of total skeletal lead burden, anticipated variations or error, and dependence of these factors on age and sex of the sampled population.

Pathophysiology of cold exposure.

The change in core temperature that occurs as a result of exposure to cold air or water affects all body systems. A decrease in core temperature induces shivering, then muscle stiffness; depresses the central nervous and respiratory systems; triggers cardiac arrhythmias and vasoconstriction; and affects body [figure: see text] fluid balance. Cold water immersion presents additional considerations, including the general shock effect on the cardiac and respiratory systems and the triggering of the diving reflex when the entire body is immersed. Basic education and precautions can prevent most cases [figure: see text] of accidental hypothermia; but when they do occur, treatment should include controlled rewarming, especially if the core temperature is below 32 degrees C.

Influence of genetic obesity, food intake and adrenalectomy in mice on selected trace element-dependent protective enzymes.

Experiments were conducted to determine if food intake and adrenalectomy influenced abnormal antioxidant defense mechanisms observed in obese mice. Paired male C57BL/6J mice of two genotypes, obese (ob/ob) and lean (+/?), were fed a nonpurified diet ad libitum or restricted (2.5 g/d) until 3 mo old. Obese mice had larger livers and kidneys but smaller brains than lean mice. Plasma ceruloplasmin activity of obese mice was 240% of that of lean mice. Restricting food intake but not adrenalectomy reduced this difference, but ceruloplasmin activity of obese mice was still 150% of that of restricted-fed lean mice. Glutathione peroxidase (GSH-Px) activity in liver of obese mice was 70% of that in control lean mice; however, in kidney GSH-Px activity was 135% of that in obese mice. Both liver and kidney GSH-Px differences were eliminated by food restriction but not by adrenalectomy. Blood and brain GSH-Px activity was not influenced by the mutation. Liver and kidney copper-zinc superoxide dismutase activity was lower in obese mice than in lean littermates, 30 and 20%, respectively. Food restriction eliminated this difference in liver but not in kidney. Glutathione S-transferase activity using 1-chloro-2,4-dinitrobenzene as substrate was 55% lower in liver (not kidney) of obese mice than in lean mice and this difference was not markedly influenced by food restriction. Obese mice have marked changes in the steady-state activities of a number of protective enzymes that are organ dependent and, in part, due to the hyperphagia associated with this mutation.

Phenol red as volume marker in rat stomach with mucosal damage.

Phenol red, long considered an acceptable volume marker in gastrointestinal experiments, has been tested in rat stomach damaged by exposure to aspirin. Significantly more phenol red was lost from stomachs damaged by aspirin than from control stomachs. Phenol red, therefore, should not be used as a volume marker in damaged stomachs. Volumes of fluid in the stomachs, which increased during the experiment, were calculated from inulin concentrations. Changes in volume of stomach fluid in experimental animals were not significantly different from those in controls.

Intragastric inulin as a measure of mucosal damage caused by aspirin.

In an attempt to find a method of gastric mucosal damage assessment that yields consistent results, the experiments presented here employed the measurement of the movement of inulin out of the gastric contents into the stomach wall and vascular compartment as an estimate of mucosal damage. Anesthetized male Sprague-Dawley rats were functionally nephrectomized and were administered a control or test solution containing 3H-inulin. The test solutions contained one of three doses of aspirin. Blood samples were taken at 15-min intervals over a 90-min exposure period. The stomach was removed from the animal and full-thickness tissue samples taken for measurement of 3H-inulin content. When the gastric mucosa was exposed to the test agents, there was a significantly greater accumulation of inulin in the body and antrum as well as in the plasma when compared to controls. We conclude that intragastric inulin can be employed to estimate gastric mucosal damage.

Cardiovascular responses to face immersion (the diving reflex) in human beings after alcohol consumption.

The diving reflex is a complex cardiovascular-respiratory response to immersion. This multireflex response in animals consists of apnea; bradycardia (the experimental parameter most often measured); peripheral vasoconstriction (resulting in preferential perfusion of the brain and heart); decreased cardiac output (with a maintained or increased stroke volume); and an increase in mean arterial blood pressure. These cardiovascular alterations act in a manner that conserves oxygen for the heart and brain. Because alcohol is often involved in water accidents with associated submersions, these initial experiments dealt with human volunteers who consumed alcohol to a blood level of 0.1 g% (legal level of intoxication). The diving reflex was elicited before and after alcohol consumption. Heart rate, blood pressure, and cardiac output were measured before, during, and after face immersion in 4 C water. In the sober state, immersion decreased heart rate, increased stroke volume, increased blood pressure, and increased total peripheral resistance. In comparison to the sober state, alcohol consumption increased pre-immersion heart rate, but did not significantly alter pre-immersion blood pressure or stroke volume. Following alcohol consumption, facial immersion decreased heart rate, but not to the same low rate as in the sober state. Alcohol consumption also evoked a lesser increase in blood pressure and failed to increase stroke volume significantly.

Human studies concerning thermal-induced shivering, postoperative "shivering," and cold-induced vasodilation.

Human reaction to cold stress and hypothermia involves shivering. Another form of overt shaking, postoperative shivering, has been attributed as a thermoregulatory response to postoperative hypothermia. Analysis of the normal human shivering pattern showed a synchronized, slow amplitude modulation (six to eight cycles/min) over all muscles sampled. In addition, there was a frequency of 8 to 10 Hz associated with each low-frequency amplitude modulation. EMG signals from postoperative patients revealed none of the major patterns seen in thermal-induced shivering. Cold-induced vasodilation also was studied and found to occur simultaneously in all cold-stressed fingers regardless of size or innervation. Thermal shivering and cold-induced vasodilation are considered to be manifestations of central neural oscillators.

Lead in bone II: skeletal-lead content as an indicator of lifetime lead ingestion and the social correlates in an archaeological population.

Measurements of skeletal-lead content (by atomic absorption spectroscopy) were made for 16 individuals recovered from a Colonial (1670-1730) plantation cemetery in Virginia. Archaeological and historical evidence allowed the identification of two social groups (plantation proprietors and laborers) within this small population, each with vastly different estimated lifetime lead exposure, reflecting different living conditions. Measured bone-lead levels confirmed these differences. The character of plantation social organization proved a more important determinant of skeletal-lead content in the individuals studied than age, sex or race.