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Fluctuation in the physical environment as a mechanism for reinforcing evolutionary transitions.

We hypothesize a mechanism for reinforcing transitions between levels of selection, involving physiological homeostasis and amplification of variation in the physical environment. Groups experience a stronger selection pressure than individuals for homeostasis with respect to reproductively limiting variables, because their greater longevity exposes them more often to suboptimal physical conditions, and greater physical size means they encompass a larger fraction of any resource/nutrient gradient. Groups achieve homeostasis by differentiation into microcosms with specialist functions, e.g. cell types. Such differentiation is more limited in individuals due to their smaller size and shorter lifespan. Hence tolerance of fluctuation in certain physical variables is proposed to be weaker in individuals than in groups. We show that a trait providing increased tolerance (alpha) to fluctuation (V-V(opt)) in a limiting abiotic variable (V), at relative fitness cost (C), can increase from rarity if the condition alpha.mid R:V-V(opt)|>C is met. Groups also sequester larger absolute quantities of resource than individuals, and group death is less frequent, hence the population dynamics of groups cause resource/nutrient availability to fluctuate with greater amplitude than that of individuals. Increasing the amplitude of fluctuation in a reproductively limiting environmental variable is proposed as a mechanism by which a group can limit reproduction of parasitic "cheat" individuals. Enhancing physical fluctuation is frequency dependent, hence only an increase in tolerance to fluctuation can explain the group's increase from rarity. However, once groups reach intermediate frequencies, a positive feedback process can be initiated in which a differentiated group enhances physical fluctuation beyond the tolerance of any "cheat", and in so doing enhances the selection pressure it experiences for homeostasis. This may help explain the persistence of transitions in individuality, and the coincidence of some such transitions with periods of change and oscillation in global scale environmental variables.

Altruism↗

Evolution of a small-muscle polymorphism in lines of house mice selected for high activity levels.

To study the correlated evolution of locomotor behavior and exercise physiology, we conducted an artificial selection experiment. From the outbred Hsd:ICR strain of Mus domesticus, we began eight separate lines, each consisting of 10 breeding pairs. In four of the lines, we used within-family selection to increase voluntary wheel running. The remaining four lines were random-bred (within lines) to serve as controls. Various traits have been monitored to test for correlated responses. Here, we report on organ masses, with emphasis on the triceps surae muscle complex, an important extensor of the ankle. Mice from the selected lines exhibit reduced total body mass, increased relative (mass-corrected) kidney mass, and reduced relative triceps surae mass. In addition, a discrete muscle-mass polymorphism was observed: some individuals had triceps surae that were almost 50% lighter than normal for their body mass. This small-muscle phenotype was observed in only three of the eight lines: in one control line, it has fluctuated in frequency between zero and 10%, whereas in two of the selected lines it has increased in frequency to approximately 50% by generation 22. Data from a set of parents and offspring (generations 23 and 24) are consistent with inheritance as a single autosomal recessive allele. Evidence for the adaptive significance of the small-muscle allele was obtained by fitting multiple-generation data to hierarchical models that include effects of genetic drift and/or selection. The small-muscle allele is estimated to have been present at low frequency (approximately 7%) in the base population, and analysis indicates that strong selection favors the allele in the selected but not control lines. We hypothesize that the small muscles possess functional characteristics and/or that the underlying allele causes pleiotropic effects (e.g., reduced total body mass; increased relative heart, liver, and kidney mass) that facilitate high levels of wheel running. Nevertheless, at generation 22, wheel running of affected individuals did not differ significantly from those with normal-sized muscles, and the magnitude of response to selection has been similar in all four selected lines, indicating that multiple genetic "solutions" are possible in response to selection for high activity levels.

Animals↗

[High dosage therapy of Parkinson disease. New dopamine agonist pramipexol allows reduction of L-dopa dose down to zero].

We report a 63-years old patient with Parkinson's disease, who experienced slight fluctuations 10 years after first signs of the disease and two years after initiation of levodopa/carbidopa and biperiden therapy. He was enrolled in a double blind, placebo controlled study with the dopamine agonist pramipexole. This study was extended to an open phase. During this phase we were able to reduce levodopa/carbidopa down to zero, while fluctuations and peak dose dyskinesias ceased. The most important adverse event were hypersomnia and very slight persistent dyskinesias, which were reported tolerable by the patient. Treatment without levodopa/carbidopa could be maintained for 31 months. We conclude that in selected cases patients in Hoehn and Yahrs stages II to III and with mild fluctuations might be treated with pramipexole without levodopa.

Antiparkinson Agents↗

The maintenance of genetic polymorphism in small island populations: large mammals in the Hebrides.

Conventionally, small populations living on islands are expected to lose genetic variation by drift. Fluctuations in population size, combined with polygynous mating systems, are expected to contribute to the process by increasing sampling effects on genetic variation. However, in individually monitored populations of Red deer on Rum and Soay sheep on St. Kilda, which experience fluctuations in population size, two processes have been identified which mitigate loss of genetic variation. First, in a number of examples, population reductions are associated with selection. Selection may be in favour of heterozygotes, or, as we have documented in several cases, it may fluctuate in direction temporally. Second, in Soay sheep, in which mortality over population crashes is male-biased, ostensibly leading to low effective numbers of males, molecular studies show that there are systematic changes in the reproductive success of young males, and in variance in male success, that broaden genetic representation compared with expectation.

Adenosine Deaminase↗

Occurrence of leu+ revertants under starvation cultures in Escherichia coli is growth-dependent.

BACKGROUND: Many investigations have reported that advantageous mutations occurred more frequently under selective conditions than those under non-selective conditions. This phenomenon is referred to as adaptive mutation. Their characteristics are that adaptive mutations are directed and growth-independent. The idea of directed adaptive mutation had been objected by some reports, however, the idea of growth-independent adaptive mutation has been held till today. RESULTS: In this paper, we have observed that under leucine starvation conditions, leu+ revertants accumulated as a function of time; leu- to leu+ reverse mutation rates and frequencies were higher than those under non starvation conditions; and no divided cells could be monitored by the penicillin method. These results were similar to the time-dependent manner of adaptive mutation from previous reports. However, leucine concentration determinate experiments revealed that certain traces of leucine, which leaked from the E. coli cells, was almost always present in the culture. More numbers of leu+ revertants appeared when the similar cultures were dropped in small areas on the selective plates than when spread on the whole selective plates. These results have shown that mutations under leucine starving conditions are growth-dependent. Fluctuation analysis of leu+ revertants indicated that leu-leu+ mutation occurred spontaneously and randomly. In addition, the spectra of leuB gene in the revertants proved that mutations under selective conditions were not specific or directed. CONCLUSIONS: The above investigations led to the conclusion (1) that the occurrence of leu+ mutations under starvation conditions was growth-dependent. The occurrence mutations was also similar to that under non-starvation conditions (2). Under starvation conditions the mutation rates were higher, and was not constant during the long process.

3-Isopropylmalate Dehydrogenase↗

Extending the coalescent to multilocus systems: the case of balancing selection.

Natural populations are structured spatially into local populations and genetically into diverse 'genetic backgrounds' defined by different combinations of selected alleles. If selection maintains genetic backgrounds at constant frequency then neutral diversity is enhanced. By contrast, if background frequencies fluctuate then diversity is reduced. Provided that the population size of each background is large enough, these effects can be described by the structured coalescent process. Almost all the extant results based on the coalescent deal with a single selected locus. Yet we know that very large numbers of genes are under selection and that any substantial effects are likely to be due to the cumulative effects of many loci. Here, we set up a general framework for the extension of the coalescent to multilocus scenarios and we use it to study the simplest model, where strong balancing selection acting on a set of n loci maintains 2n backgrounds at constant frequencies and at linkage equilibrium. Analytical results show that the expected linked neutral diversity increases exponentially with the number of selected loci and can become extremely large. However, simulation results reveal that the structured coalescent approach breaks down when the number of backgrounds approaches the population size, because of stochastic fluctuations in background frequencies. A new method is needed to extend the structured coalescent to cases with large numbers of backgrounds.

Alleles↗

Leptin increases energy expenditure and selectively promotes fat metabolism in ob/ob mice.

Obesity occurs whenever energy intake exceeds energy expenditure. The ob gene product leptin is a potent anorectic agent when administered to ob/ob mice, but its effects on energy expenditure have not been investigated in detail. The present study was designed to analyze the acute metabolic effects of leptin in vivo. Analysis of oxygen consumption in ob/ob mice demonstrated a reduction in energy expenditure compared with lean controls; this reduction showed a diurnal fluctuation and was most evident during the light cycle. A single intraperitoneal dose of leptin increased oxygen consumption during the light cycle in ob/ob mice, ablating the circadian fluctuation in this parameter. In addition, leptin had a profound effect on fuel selection: the respiratory quotient was markedly reduced, indicating a reduction in carbohydrate oxidation and an increase in fat oxidation. These acute effects of leptin on metabolic parameters are consistent with the selective loss of body fat observed on chronic leptin treatment and suggest that increased energy utilization plays an important role in the anti-obese actions of leptin.

Analysis of Variance↗

Reduction in intraventricular hemorrhage by elimination of fluctuating cerebral blood-flow velocity in preterm infants with respiratory distress syndrome.

In a previous study of preterm infants requiring mechanical ventilation for the respiratory distress syndrome, we demonstrated a striking association of fluctuating cerebral blood-flow velocity in the first day of life with the subsequent occurrence of intraventricular hemorrhage. Because this fluctuating pattern could be eliminated by muscle paralysis, we conducted a prospective study of preterm infants receiving mechanical ventilation for the respiratory distress syndrome in which we evaluated the effect of paralysis and this flow-velocity pattern on the incidence and severity of intraventricular hemorrhage. Twenty-four infants with the fluctuating pattern in the first hours of life were identified and randomly selected to serve as controls (10) or to be subjected to muscle paralysis (14). Intraventricular hemorrhage developed in all 10 control infants but in only 5 of the 14 infants subjected to muscle paralysis. Moreover, in 4 of the 5 paralyzed infants in whom hemorrhage developed, it did so after cessation of the paralysis. Seven of the 10 control infants had Grade III hemorrhage, the most severe variety of intraventricular hemorrhage, whereas none of the paralyzed infants had Grade III hemorrhage. We conclude that elimination of fluctuating cerebral blood-flow velocity in preterm infants with respiratory distress syndrome markedly reduces the incidence and severity of intraventricular hemorrhage.

Blood Flow Velocity↗

Interaction between selection and genetic drift in laboratory populations of Drosophila bipectinata.

To study the effect of selection and random genetic drift in laboratory populations of D. bipectinata, cut wing mutant and wild type stocks were crossed in several bottles. All the lines were maintained for several generations and in each generation the number of wild type and mutant flies was scored. Based on the number of two types of males, the frequency of cut gene and its normal allele was calculated for different bottles since ct is a sex-linked mutation and males are hemizygous. Interestingly, the cut gene was quickly eliminated in all the populations which is due to selection affecting fitness of two types of flies. However, there was considerable fluctuation in gene frequency among different populations and also there was increase in the cut gene frequency in certain generations as compared to previous generation which is caused due to effect of genetic drift which operates in small populations.

Animals↗

Rapid feature selective neuronal synchronization through correlated latency shifting.

Spontaneous brain activity could affect processing if it were structured. We show that neuron pairs in cat primary visual cortex exhibited correlated fluctuations in response latency, particularly when they had overlapping receptive fields or similar orientation preferences. Correlations occurred within and across hemispheres, but only when local field potentials (LFPs) oscillated in the gamma-frequency range (40-70 Hz). In this range, LFP fluctuations preceding response onset predicted response latencies; negative (positive) LFPs were associated with early (late) responses. Oscillations below 10 Hz caused covariations in response amplitude, but exhibited no columnar selectivity or coordinating effect on latencies. Thus, during high gamma activity, spontaneous activity exhibits distinct, column-specific correlation patterns. Consequently, cortical cells undergo coherent fluctuations in excitability that enhance temporal coherence of responses to contours that are spatially contiguous or have similar orientation. Because synchronized responses are more likely than dispersed responses to undergo rapid and joint processing, spontaneous activity may be important in early visual processes.

Action Potentials↗

Geographical variation in the rate of evolution: effect of available energy or fluctuating environment?

In a recent paper, Wright et al. (2003) argue for the hypothesis that greater biologically available energy elevates the rate of molecular evolution. However, their results are also consistent with alternative hypotheses that invoke either environmentally driven variation in effective population sizes, or natural selection, or both. The available energy gradient cited by Wright et al. is linearly correlated with temperature fluctuations, and the observed rate heterogeneity could be a consequence of this environmental variability. The distribution of phylogenetic branch lengths alone is insufficient to distinguish between the hypotheses, and complementary approaches are suggested.

DNA, Plant↗

Decreased mutation rate for cellular resistance to doxorubicin and suppression of mdr1 gene activation by the cyclosporin PSC 833.

BACKGROUND: Various mechanisms can contribute to cellular resistance to doxorubicin. These include expression of the multidrug transporter P-glycoprotein (product of the mdr1 gene [also known as PGY1], Mrp (multidrug resistance-associated protein), the p110 major vault protein, altered glutathione metabolism, and altered levels or activity of topoisomerase II (Topo II). We reported recently that single-step treatment of human MES-SA sarcoma cells with 40 nM doxorubicin resulted in selection of spontaneous mutants at a rate of 1.8 x 10(-6) per cell generation. All individually selected mutants manifested the multidrug-resistant phenotype, related to activation of the mdr1 gene. PURPOSE: Luria and Delbrück fluctuation analysis was performed with MES-SA cells to determine the mutation rate and the nature and mechanisms of resistance after single-step selection with doxorubicin in the presence of the cyclosporin PSC 833, a potent modulator of multidrug resistance. METHODS: Ten flasks were seeded with 2000 cells/flask and grown to confluent populations of approximately 8 x 10(6) cells. After reseeding in 96-well plates, the populations were treated with 40 nM doxorubicin and 2 microM PSC 833 for 3 weeks. Surviving colonies were scored, individually harvested, and propagated. The drug-resistant phenotype was assessed by the tetrazolium dye (MTT) cytotoxicity assay and by monitoring cellular glutathione content and radiolabeled drug accumulation. Coupled reverse transcription-polymerase chain reaction (RT-PCR) was used to evaluate mdr1, MRP, Topo II alpha, and Topo II beta gene expression. Topo II, P-glycoprotein, and p110 levels were examined by immunoblotting or immunocytochemistry. Topo II activity was assessed by decatenation of kinetoplast DNA, and etoposide-induced cleavable complex formation was studied by the potassium-sodium dodecyl sulfate precipitation assay. RESULTS: Mutations were detected at a rate of 2.5 x 10(-7) per cell generation. Analysis of variance indicates that spontaneous mutations, rather than changes in cellular function, conferred resistance to doxorubicin and PSC 833. None of the isolated clones expressed mdr1 messenger RNA or P-glycoprotein, and none exhibited an increase in MRP expression. No alterations were found in cellular glutathione content, intracellular accumulations of daunorubicin and etoposide, levels of p110 protein, or levels of Topo II beta transcripts. However, a significant decrease in Topo II alpha messenger RNA and protein was found in all examined clones, as well as decreased Topo II catalytic activity and reduced cleavable complex formation in the presence of etoposide. CONCLUSIONS: PSC 833 co-selection reduced the mutation rate for doxorubicin-selected resistance by 10-fold and suppressed the emergence of mdr1 mutants. Survival of cells exposed to doxorubicin and PSC 833 occurs by selection of spontaneously arising mutants that exhibit altered Topo II alpha expression. IMPLICATIONS: Our results suggest that treatment with multidrug resistance modulators such as PSC 833 together with multidrug resistance-related cytotoxins may suppress the activation of mdr1 and prevent the emergence of resistant cancer cell clones with the multidrug-resistant phenotype.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Investigations into neuropeptide Y-mediated presynaptic inhibition in cultured hippocampal neurones of the rat.

1. We have examined the effects of neuropeptide Y (NPY) on synaptic transmission and [Ca2+]i signals in rat hippocampal neurones grown in culture. [Ca2+]i in individual neurones displayed frequent spontaneous fluctuations often resulting in an elevated plateau [Ca2+]i. These fluctuations were reduced by tetrodotoxin (1 microM) or combinations of the excitatory amino acid antagonists 6-cyano-7-dinitro-quinoxaline (CNQX) (10 microM) and aminophosphonovalerate (APV) (50 microM), indicating that they were the result of glutamatergic transmission occurring between hippocampal neurones. 2. [Ca2+]i fluctuations were also prevented by Ni2+ (200 microM), by the GABAB receptor agonist, baclofen (10 microM) and by NPY (100 nM) or Y2 receptor-selective NPY agonists. Following treatment of cells with pertussis toxin, NPY produced only a brief decrease in [Ca2+]i fluctuations which rapidly recovered. 3. Perfusion of hippocampal neurones with 50 mM K+ produced a large rapid increase in [Ca2+]i. This increase was slightly reduced by NPY or by a combination of CNQX and APV. The effects of CNQX/APV occluded those of NPY. NPY had no effect on Ba2+ currents measured in hippocampal neurones under whole cell voltage-clamp even in the presence of intracellular GTP-gamma-S. On the other hand, Ba2+ currents were reduced by both Cd2+ (200 microM) and baclofen (10 microM). 4. Current clamp recordings from hippocampal neurones demonstrated the occurrence of spontaneous e.p.s.ps and action potential firing which were accompanied by increases in [Ca2+]i. This spontaneous activity and the accompanying [Ca2+]i signals were prevented by application of NPY (100 nM). When hippocampal neurones were induced to fire trains of action potentials in the absence of synaptic transmission, these were accompanied by an increase in cell soma [Ca2+]j. NPY (100 nM) had no effect on these cell soma [Ca2+], signals. NPY (100 nM) also had no effect on inward currents generated in hippocampal neurones by micropipette application of glutamate (50 microM).5. Thus, NPY is able to abolish excitatory neurotransmission in hippocampal cultures through a pertussis toxin-sensitive mechanism. However, no effect of NPY on Ca2+ influx into the cell soma of these hippocampal neurones could be discerned. These results are consistent with a localized presynaptic inhibitory effect of NPY on glutamate release in hippocampal neurones in culture.

2-Amino-5-phosphonovalerate↗

Screening for micro-albuminuria with Micro-Bumintest tablets and albumin/creatinine ratio.

The value of Micro-Bumintest tablets and the albumin/creatinine ratio in first morning (FM) urine samples to screen for micro-albuminuria in diabetic patients was evaluated. The A/C ratio in FM urine was highly correlated with the urinary albumin excretion (UAE) rate in micrograms/min in timed overnight (0) urine (R = 0.95, p < 0.001). The sensitivity, specificity and positive and negative predictive values of an A/C ratio > 3.0 to predict micro-albuminuria defined as an UAE > 20 micrograms/min were 94, 92, 92 and 94% respectively. The day-to-day fluctuation of albumin excretion varied between 10 and 105%. The sensitivity, specificity and positive and negative predictive values of micro-bumintest tablet reactions on FM urine samples to predict an A/C ratio > 3.0 were 93, 59, 63 and 91% respectively. It is concluded that for screening of micro-albuminuria the A/C ratio in FM urine is as precise as timed 0 urine collection, but with more comfort for the patient. Micro-Bumintest tablets are useful as an initial screening test and will decrease the number of quantitative albumin measurements in the screening programme by about 30% if only urine samples with positive tablet test are selected for a further investigation. Because of considerable day-to-day fluctuation of urinary albumin excretion several urine samples should be investigated before presence of incipient diabetic nephropathy is assumed.

Albuminuria↗

Coordination topology and stability for the native and binding conformers of chymotrypsin inhibitor 2.

We demonstrate that the stabilization of the binding region is accomplished at the expense of a loss in the stability of the rest of the protein. A novel molecular mechanics (MM) approach is introduced to distinguish residue stabilities of proteins in a given conformation. As an example, the relative stabilities of folded chymotrypsin inhibitor 2 (CI2) in unbound form, and CI2 in complex with subtilisin novo is investigated. The conformation of the molecule in the two states is almost identical, with an approximately 0.6-A root-mean-square deviation (RMSD) of the Calpha atoms. On binding, the packing density changes only at the binding loop. However, residue fluctuations in the rest of the protein are greatly altered solely due to those contacts, indicating the effective propagation of perturbation and the presence of remotely controlling residues. To quantify the interplay between packing density, packing order, residue fluctuations, and residue stability, we adopt an MM approach whereby small displacements are inserted at selected residues, followed by energy minimization; the displacement of each residue in response to such perturbations are organized in a perturbation-response matrix L. We define residue stability lambda(i) = summation operator((j)L(ij))/ summation operator((j) L(ji)) as the ratio of the amount of change to which the residue is amenable, to the ability of a given residue to induce change. We then define the free energy associated with residue stability, DeltaG(lambda) = -RT ln lambda. DeltaG(lambda) intrinsically selects the residues that are in the folding core. Upon complexation, the binding loop becomes more resistant to perturbation, in contrast to the alpha-helix that favors change. Although the two forms of CI2 are structurally similar, residue fluctuations differ vastly, and the stability of many residues is altered upon binding. The decrease in entropy introduced by binding is thus compensated by these changes.

Biomechanical Phenomena↗

Effects of reproductive compensation and genetic drift on X-linked lethals.

A revival of interest in Haldane's equilibrium theory for X-linked lethals has been stimulated by the introduction of accurate tests for the detection of female heterozygotes in Lesch-Nyhan disease. Application of these tests appears to indicate an excess of familial cases. This excess can be attributed to ascertainment bias, a difference in female and male mutation rates, genetic drift, and reproductive compensation. Reproductive compensation will be particularly effective in increasing the proportion of familial cases if (1) birth control is widespread; (2) selection against affected males acts in utero; (3) affected sons show symptoms at an early age; and (4) sons are more highly valued than daughters. We demonstrate how only a few generations of reproductive compensation are sufficient to achieve an approximate equilibrium between selection and mutation showing a high proportion of familial cases. We also discuss the random fluctuations around equilibrium caused by genetic drift.

Contraception↗

Anti-Müllerian hormone levels in the spontaneous menstrual cycle do not show substantial fluctuation.

CONTEXT: Anti-Müllerian hormone (AMH), a quantitative marker for ovarian reserve, has been suggested to be independent of the classical endocrine fluctuations of the menstrual cycle. OBJECTIVE: The objective of the study was to determine whether AMH levels are constant throughout the menstrual cycle, compared with those of FSH, LH, and estradiol. DESIGN/PATIENTS: Frequent blood sampling was performed in 44 fertile, regularly cycling, female volunteers during one full menstrual cycle. SETTING: The study was conducted at a university hospital. MAIN OUTCOME MEASURES: AMH, FSH, LH, and estradiol measurements were allocated to one of seven cycle phases, and a multilevel analysis was performed. Consistent fluctuation patterns were tested by fitting sine patterns to the data. Finally, the frequency in which randomly selected individual samples would remain in one of five preset level categories (quintiles) for each of the variables was studied. RESULTS: A sine pattern fitted to the AMH data was not statistically significant (P = 0.40). In contrast, sine patterns for FSH, LH, and estradiol were highly significant. Comparing the seven cycle phases, no significant differences could be observed between phase-specific AMH levels (P = 0.06). Repeated selection of AMH samples for each individual showed that in 71.5% of selections, AMH values remained in the same quintile, whereas in 27.9% values fell in an adjacent quintile. CONCLUSIONS: AMH levels measured through a full menstrual cycle did not show consistent fluctuation patterns in contrast to levels of FSH, LH, and estradiol. Furthermore, random fluctuations were small, indicating that AMH can be relied on as a cycle-independent marker for ovarian reserve.

Adult↗

Use of environmental TLD data at a nuclear power station to estimate detection limits for radiation exposure due to station operation.

The environmental radiation monitoring network maintained by the Georgia Department of Natural Resources at the Hatch Nuclear Plant, Baxley, GA, was used to determine the variability in measured background exposures in order to estimate the detection limits for radiation due to station operation. Data were obtained from LiF thermoluminescent dosimeters (TLDs) exposed in sets of four at 46 locations for 16 consecutive quarters from 1982 to 1985. The average exposure for 695 values was 11.8 microC kg-1 y-1 (mR/quarter). At individual locations, the averages of quarterly exposures ranged from 9.3 to 14.3 microC kg-1 y-1 and had standard deviations from 0.8 to 2.2 microC kg-1 y-1 with a typical standard deviation of 1.5 microC kg-1 y-1. The standard deviation due to reading the TLDs and subtracting the transit plus storage exposure, estimated as 0.4 microC kg-1 y-1, contributed only a small fraction of the uncertainty of the mean background; the greater contribution is from temporal fluctuations. There is a spatial pattern of higher background along the Altamaha River but no distinct seasonal pattern. The observation that exposure values at most locations rose and fell in unison suggested a calculational model that adjusts the average exposure level at a location by an increment determined each quarter at control locations. This model reduced the standard deviation for a predicted quarterly background exposure to 0.9 microC kg-1 y-1, with a range of 0.5 to 1.3 microC kg-1 y-1. If the standard deviation of the net value is multiplied by an error probability constant of 3.3 (type I and II errors not greater than 5%), the detection limits range from 2 to 5 microC kg-1 y-1 at the individual locations, with an average of 3 microC kg-1 y-1. This approach or two similar ones described earlier are recommended for improving the detection limit where needed. The standard deviation, and hence the detection limit, can be further reduced by selecting a subset of indicator and control locations with more closely matched background exposure fluctuations. This was demonstrated with five indicator and five control locations, but further study is needed to determine selection guides.

Cesium Radioisotopes↗