Investigation of competitive antagonist binding to the nicotinic acetylcholine receptor using voltage-jump and light-flash techniques

1. The effect of 2,2’-bis-[α-(trimethylammonium)methyl]azobenzene (2BQ), a photoisomerizable competitive antagonist, was studied at the nicotinic acetycholine receptor of Electrophorus electroplaques using voltage-jump and light-flash techniques.

2. 2BQ, at concentrations below 3 μΜ, reduced the amplitude of voltage-jump relaxations but had little effect on the voltage-jump relaxation time constants under all experimental conditions. At higher concentrations and voltages more negative than -150 mV, 2BQ caused significant open channel blockade.

3. Dose-ratio studies showed that the cis and trans isomers of 2BQ have equilibrium binding constants (K _ᵢ) of .33 and 1.0 μΜ, respectively. The binding constants determined for both isomers are independent of temperature, voltage, agonist concentration, and the nature of the agonist.

4. In a solution of predominantly cis-2BQ, visible-light flashes led to a net cis→trans isomerization and caused an increase in the agonist-induced current. This increase had at least two exponential components; the larger amplitude component had the same time constant as a subsequent voltage-jump relaxation; the smaller amplitude component was investigated using ultraviolet light flashes.

5. In a solution of predominantly trans-2BQ, UV-light flashes led to a net trans→cis isomerization and caused a net decrease in the agonist-induced current. This effect had at least two exponential components. The smaller and faster component was an increase in agonist-induced current and had a similar time constant to the voltage-jump relaxation. The larger component was a slow decrease in the agonist-induced current with rate constant approximately an order of magnitude less than that of the voltage-jump relaxation. This slow component provided a measure of the rate constant for dissociation of cis-2BQ (k_ = 60/s at 20°C). Simple modelling of the slope of the dose-rate curves yields an association rate constant of 1.6 x 10⁸/M/s. This agrees with the association rate constant of 1.8 x 10⁸/M/s estimated from the binding constant (K_i). The Q₁₀ of the dissociation rate constant of cis-2BQ was 3.3 between 6° and 20°C. The rate constants for association and dissociation of cis-28Q at receptors are independent of voltage, agonist concentration, and the nature of the agonist.

6. We have measured the molecular rate constants of a competitive antagonist which has roughly the same K _ᵢ as d-tubocurarine but interacts more slowly with the receptor. This leads to the conclusion that curare itself has an association rate constant of 4 x 10⁹/M/s or roughly as fast as possible for an encounter-limited reaction.

A model for energy and morphology of crystalline grain boundaries with arbitrary geometric character

It has been well-established that interfaces in crystalline materials are key players in the mechanics of a variety of mesoscopic processes such as solidification, recrystallization, grain boundary migration, and severe plastic deformation. In particular, interfaces with complex morphologies have been observed to play a crucial role in many micromechanical phenomena such as grain boundary migration, stability, and twinning. Interfaces are a unique type of material defect in that they demonstrate a breadth of behavior and characteristics eluding simplified descriptions. Indeed, modeling the complex and diverse behavior of interfaces is still an active area of research, and to the author's knowledge there are as yet no predictive models for the energy and morphology of interfaces with arbitrary character. The aim of this thesis is to develop a novel model for interface energy and morphology that i) provides accurate results (especially regarding "energy cusp" locations) for interfaces with arbitrary character, ii) depends on a small set of material parameters, and iii) is fast enough to incorporate into large scale simulations.

In the first half of the work, a model for planar, immiscible grain boundary is formulated. By building on the assumption that anisotropic grain boundary energetics are dominated by geometry and crystallography, a construction on lattice density functions (referred to as "covariance") is introduced that provides a geometric measure of the order of an interface. Covariance forms the basis for a fully general model of the energy of a planar interface, and it is demonstrated by comparison with a wide selection of molecular dynamics energy data for FCC and BCC tilt and twist boundaries that the model accurately reproduces the energy landscape using only three material parameters. It is observed that the planar constraint on the model is, in some cases, over-restrictive; this motivates an extension of the model.

In the second half of the work, the theory of faceting in interfaces is developed and applied to the planar interface model for grain boundaries. Building on previous work in mathematics and materials science, an algorithm is formulated that returns the minimal possible energy attainable by relaxation and the corresponding relaxed morphology for a given planar energy model. It is shown that the relaxation significantly improves the energy results of the planar covariance model for FCC and BCC tilt and twist boundaries. The ability of the model to accurately predict faceting patterns is demonstrated by comparison to molecular dynamics energy data and experimental morphological observation for asymmetric tilt grain boundaries. It is also demonstrated that by varying the temperature in the planar covariance model, it is possible to reproduce a priori the experimentally observed effects of temperature on facet formation.

Finally, the range and scope of the covariance and relaxation models, having been demonstrated by means of extensive MD and experimental comparison, future applications and implementations of the model are explored.

Formal methods in the foundations of science

The intent of this study is to provide formal apparatus which facilitates the investigation of problems in the methodology of science. The introduction contains several examples of such problems and motivates the subsequent formalism.

A general definition of a formal language is presented, and this definition is used to characterize an individual’s view of the world around him. A notion of empirical observation is developed which is independent of language. The interplay of formal language and observation is taken as the central theme. The process of science is conceived as the finding of that formal language that best expresses the available experimental evidence.

To characterize the manner in which a formal language imposes structure on its universe of discourse, the fundamental concepts of elements and states of a formal language are introduced. Using these, the notion of a basis for a formal language is developed as a collection of minimal states distinguishable within the language. The relation of these concepts to those of model theory is discussed.

An a priori probability defined on sets of observations is postulated as a reflection of an individual’s ontology. This probability, in conjunction with a formal language and a basis for that language, induces a subjective probability describing an individual’s conceptual view of admissible configurations of the universe. As a function of this subjective probability, and consequently of language, a measure of the informativeness of empirical observations is introduced and is shown to be intuitively plausible – particularly in the case of scientific experimentation.

The developed formalism is then systematically applied to the general problems presented in the introduction. The relationship of scientific theories to empirical observations is discussed and the need for certain tacit, unstatable knowledge is shown to be necessary to fully comprehend the meaning of realistic theories. The idea that many common concepts can be specified only by drawing on knowledge obtained from an infinite number of observations is presented, and the problems of reductionism are examined in this context.

A definition of when one formal language can be considered to be more expressive than another is presented, and the change in the informativeness of an observation as language changes is investigated. In this regard it is shown that the information inherent in an observation may decrease for a more expressive language.

The general problem of induction and its relation to the scientific method are discussed. Two hypotheses concerning an individual’s selection of an optimal language for a particular domain of discourse are presented and specific examples from the introduction are examined.

Influence of temperature and salinity fluctuations on propagation behaviour of partially coherent beams in oceanic turbulence

A theoretical study of the behaviour of partially coherent beams propagating through oceanic turbulence has been performed. Based on the previously developed knowledge of beam spreading of a partially coherent beam in the atmosphere and the spatial power spectrum of the refractive index of ocean water, we study the normalized root-mean-square width of a partially coherent beam on propagation through oceanic turbulence and its turbulence distance which may be a measure of turbulence resistance. Our analysis indicates that the behaviour of partially coherent beams on propagation may be described by the rate of dissipation of the mean-squared temperature chi(T) and that of salinity chi(S). In terms of a quantity w that defines the contributions of the temperature and salinity distributions to the distribution of the refractive index, chi(S) could be written as a function of chi(T) and w. Therefore, the behaviour of partially coherent beams on propagation can be characterized only by chi(T) for a given w. The results are shown for curved surfaces, from which one can see that partially coherent beams exhibit robust turbulence resistance when the water volume has a smaller chi(T).

An analysis of migratory salmonid catch effort data, derived from anglers' log books, 1991

1000 log books were issued to anglers of which 236 were returned, those from the rivers Derwent, Kent, Lune and Ribble accounted for the vast majority. The Derwent had the highest catch rate of these rivers: one salmon every 13.89 hours followed by the Lune, Kent and Ribble at 16.39, 18.87 and 35.71 hours, respectively. For sea trout the Lune, Derwent and Ribble had a catch rate of approximately one fish every 10.0 hours (9.8, 10.0 and 10.64 hours),and for the Kent one fish per 16.1 hours fished. Salmon angling visits were, in general,longer than those for sea trout being between 2 and 6 hours as opposed to 2 to 4 hours. On the majority of visits (>80%) no fish were caught and was the same for salmon and sea trout. For salmon the majority of fish were caught on fly, spinner or worm, and the least on prawn. For sea trout fly predominated. The majority of salmon caught were less than 91b in weight and were presumed to be grilse (1 sea winter). The majority of the sea trout caught weighed between 1 and 31b. The pattern of catch, effort, CPUE, abundance and catchability for salmon and sea trout were modelled using the data from the rivers Derwent, Kent and Lune. Flow significantly influenced catch, effort and catchability of salmon which had entered in a particular month. For sea trout flow was not significantly correlated with any of the dependent variables. The catchability coefficient for salmon, determined from the total number of fish, remained relatively constant over the period June to October indicating that CPUE was a reasonable measure of within season abundance. This was not found to be the case for sea trout.

Energy analysis of the ring seine operations, off Cochin, Kerala

Ring seines are lightly constructed purse seines adapted for operation in the traditional sector. Fish production and energy requirement in the ring seine operations, off Cochin, Kerala, India are discussed in this paper, based on data collected during 1997- 1998. The results reflect the Gross Energy Requirement (GER) situation that existed during 1997-1998. Mean catch per ring seiner per year worked out to be 211.9 t of which sardines (Sardinella spp.) constituted 44.3%, followed by Indian mackerel (Rastrelliger kanagurta) 29.7%, carangids 11.4%, penaeid prawns 2.2%, pomfrets 1.1% and miscellaneous fish 11.3%. Total energy inputs into the ring seine operations were estimated to be 1300.8 GJ. Output by way of fish production was determined to be 931.85 GJ. GER is the sum of all non-renewable energy resources consumed in making available a product or service and is a measure of intensity of non-renewable resource use. GER per tonne of fish landed by ring seiners was estimated to be 6.14. Among the operational inputs, kerosene constituted 73.4% of the GER, followed by petrol (12.7%), diesel (6.7%) and lubricating oil (2.4%). Fishing gear contributed 3.8%, engine 0.8% and fishing craft 0.3% of the GER. Energy ratio for ring seining was 0.72 and energy intensity 1.40.

Relative pleopod length as an indicator of size at sexual maturity in slipper (Scyllarides squammosus) and spiny Hawaiian (Panulirus marginatus) lobsters

Body size at gonadal maturity is described for females of the slipper lobster (Scyllarides squammosus) (Scyllaridae) and the endemic Hawaiian spiny lobster (Panulirus marginatus) (Palinuridae) based on microscopic examination of histological preparations of ovaries. These data are used to validate several morphological metrics (relative exopodite length, ovigerous condition) of functional sexual maturity. Relative exopodite length (“pleopod length”) produced consistent estimates of size at maturity when evaluated with a newly derived statistical application for estimating size at the morphometric maturation point (MMP) for the population, identified as the midpoint of a sigmoid function spanning the estimated boundaries of overlap between the largest immature and smallest adult animals. Estimates of the MMP were related to matched (same-year) characterizations of sexual maturity based on ovigerous condition — a more conventional measure of functional maturity previously used to characterize maturity for the two lobster species. Both measures of functional maturity were similar for the respective species and were within 5% and 2% of one another for slipper and spiny lobster, respectively. The precision observed for two shipboard collection series of pleopod-length data indicated that the method is reliable and not dependent on specialized expertise. Precision of maturity estimates for S. squammosus with the pleopod-length metric was similar to that for P. marginatus with any of the other measures (including conventional evidence of ovigerous condition) and greatly exceeded the precision of estimates for S. squammosus based on ovigerous condition alone. The two measures of functional maturity averaged within 8% of the estimated size at gonadal maturity for the respective species. Appendage-to-body size proportions, such as the pleopod length metric, hold great promise, particularly for species of slipper lobsters like S. squammosus for which there exist no other reliable conventional morphological measures of sexual maturity. Morphometric proportions also should be included among the factors evaluated when assessing size at sexual maturity in spiny lobster stocks; previously, these proportions have been obtained routinely only for brachyuran crabs within the Crustacea.

Simulation of Tail Weight Distributions in Biological Year 1986–2006 Landings of Brown Shrimp, Farfantepenaeus aztecus, from the Northern Gulf of Mexico Fishery

Size distribution within re- ported landings is an important aspect of northern Gulf of Mexico penaeid shrimp stock assessments. It reflects shrimp population characteristics such as numerical abundance of various sizes, age structure, and vital rates (e.g. recruitment, growth, and mortality), as well as effects of fishing, fishing power, fishing practices, sampling, size-grading, etc. The usual measure of shrimp size in archived landings data is count (C) the number of shrimp tails (abdomen or edible portion) per pound (0.4536 kg). Shrimp are marketed and landings reported in pounds within tail count categories. Statistically, these count categories are count class intervals or bins with upper and lower limits expressed in C. Count categories vary in width, overlap, and frequency of occurrence within the landings. The upper and lower limits of most count class intervals can be transformed to lower and upper limits (respectively) of class intervals expressed in pounds per shrimp tail, w, the reciprocal of C (i.e. w = 1/C). Age based stock assessments have relied on various algorithms to estimate numbers of shrimp from pounds landed within count categories. These algorithms required un- derlying explicit or implicit assumptions about the distribution of C or w. However, no attempts were made to assess the actual distribution of C or w. Therefore, validity of the algorithms and assumptions could not be determined. When different algorithms were applied to landings within the same size categories, they produced different estimates of numbers of shrimp. This paper demonstrates a method of simulating the distribution of w in reported biological year landings of shrimp. We used, as examples, landings of brown shrimp, Farfantepenaeus aztecus, from the northern Gulf of Mexico fishery in biological years 1986–2006. Brown shrimp biological year, Ti, is defined as beginning on 1 May of the same calendar year as Ti and ending on 30 April of the next calendar year, where subscript i is the place marker for biological year. Biological year landings encompass most if not all of the brown shrimp life cycle and life span. Simulated distributions of w reflect all factors influencing sizes of brown shrimp in the landings within a given biological year. Our method does not require a priori assumptions about the parent distributions of w or C, and it takes into account the variability in width, overlap, and frequency of occurrence of count categories within the landings. Simulated biological year distributions of w can be transformed to equivalent distributions of C. Our method may be useful in future testing of previously applied algorithms and development of new estimators based on statistical estimation theory and the underlying distribution of w or C. We also examine some applications of biological year distributions of w, and additional variables derived from them.

Patterns of orographic uplift in the Sierra Nevada and their relationship to upper-level atmospheric circulation

We examine monthly and seasonal patterns of precipitation across various elevations of the eastern Central Valley of California and the Sierra Nevada. A measure of the strength of the orographic effect called the “precipitation ratio” is calculated, and we separate months into four groups based on being wet or dry and having low or high precipitation ratios. Using monthly maps of mean 700-mb height anomalies, we describe the northern hemisphere mid-tropospheric circulation patterns associated with each of the four groups. Wet months are associated with negative height anomalies over the eastern Pacific, as expected. However, the orientation of the trough is different for years with high and low precipitation ratios. Wet months with high ratios typically have circulation patterns factoring a west-southwest to east-northeast storm track from around the Hawaiian Islands to the Pacific Northwest of the United States. Wet months with low precipitation ratios are associated with a trough centered near the Aleutians and a northwest to southeast storm track. Dry months are marked by anticyclones in the Pacific, but this feature is more localized to the eastern Pacific for months with low precipitation ratios than for those with high ratios. Using precipitation gauge and snow course data from the American River and Truckee-Tahoe basins, we determined that the strength of the orographic effect on a seasonal basis is spatially coherent at low and high elevations and on opposite sides of the Sierra Nevada crestline.

Review of size- and age-dependence in batch spawning: implications for stock assessment of fish species exhibiting indeterminate fecundity

Most assessments of fish stocks use some measure of the reproductive potential of a population, such as spawning biomass. However, the correlation between spawning biomass and reproductive potential is not always strong, and it likely is weakest in the tropics and subtropics, where species tend to exhibit indeterminate fecundity and release eggs in batches over a protracted spawning season. In such cases, computing annual reproductive output requires estimates of batch fecundity and the annual number of batches—the latter subject to spawning frequency and duration of spawning season. Batch fecundity is commonly measured by age (or size), but these other variables are not. Without the relevant data, the annual number of batches is assumed to be invariant across age. We reviewed the literature and found that this default assumption lacks empirical support because both spawning duration and spawning frequency generally increase with age or size. We demonstrate effects of this assumption on measures of reproductive value and spawning potential ratio, a metric commonly used to gauge stock status. Model applications showed substantial sensitivity to age dependence in the annual number of batches. If the annual number of batches increases with age but is incorrectly assumed to be constant, stock assessment models would tend to overestimate the biological reference points used for setting harvest rates. This study underscores the need to better understand the age- or size-dependent contrast in the annual number of batches, and we conclude that, for species without evidence to support invariance, the default assumption should be replaced with one that accounts for age- or size-dependence.

Hydrodynamic entrapment of bacteria swimming near a solid surface.

The near-surface motility of bacteria is important in the initial formation of biofilms and in many biomedical applications. The swimming motion of Escherichia coli near a solid surface is investigated both numerically and experimentally. A boundary element method is used to predict the hydrodynamic entrapment of E. coli bacteria, their trajectories, and the minimum separation of the cell from the surface. The numerical results show the existence of a stable swimming distance from the boundary that depends only on the shape of the cell body and the flagellum. The experimental validation of the numerical approach allows one to use the numerical method as a predictive tool to estimate with reasonable accuracy the near-wall motility of swimming bacteria of known geometry. The analysis of the numerical database demonstrated the existence of a correlation between the radius of curvature of the near-wall circular trajectory and the separation gap. Such correlation allows an indirect estimation of either of the two quantities by a direct measure of the other without prior knowledge of the cell geometry. This result may prove extremely important in those biomedical and technical applications in which the near-wall behavior of bacteria is of fundamental importance.

Integration of fisheries acoustics surveys and bathymetric mapping to characterize midwater-seafloor habitats of US Virgin Islands and Puerto Rico (2008-2010)

NOAA’s Coral Reef Conservation program (CRCP) develops coral reef management priorities by bringing together various partners to better understand threats to coral reef ecosystems with the goal of conserving, protecting and restoring these resources. Place-based and ecosystem-based management approaches employed by CRCP require that spatially explicit information about benthic habitats and fish utilization are available to characterize coral reef ecosystems and set conservation priorities. To accomplish this, seafloor habitat mapping of coral reefs around the U.S. Virgin Islands (USVI) and Puerto Rico has been ongoing since 2004. In 2008, fishery acoustics surveys were added to NOAA survey missions in the USVI and Puerto Rico to assess fish distribution and abundance in relation to benthic habitats in high priority conservation areas. NOAA’s National Centers for Coastal Ocean Science (NCCOS) have developed fisheries acoustics survey capabilities onboard the NOAA ship Nancy Foster to complement the CRCP seafloor habitat mapping effort spearheaded by the Center for Coastal Monitoring and Assessment Biogeography Branch (CCMA-BB). The integration of these activities has evolved on the Nancy Foster over the three years summarized in this report. A strategy for improved operations and products has emerged over that time. Not only has the concurrent operation of multibeam and fisheries acoustics surveys been beneficial in terms of optimizing ship time and resources, this joint effort has advanced an integrated approach to characterizing bottom and mid-water habitats and the fishes associated with them. CCMA conducts multibeam surveys to systematically map and characterize coral reef ecosystems, resulting in products such as high resolution bathymetric maps, backscatter information, and benthic habitat classification maps. These products focus on benthic features and live bottom habitats associated with them. NCCOS Centers (the Center for Coastal Fisheries and Habitat Research and the Center for Coastal Environmental Health and Biomolecular Research) characterize coral reef ecosystems by using fisheries acoustics methods to capture biological information through the entire water column. Spatially-explicit information on marine resources derived from fisheries acoustics surveys, such as maps of fish density, supports marine spatial planning strategies and decision making by providing a biological metric for evaluating coral reef ecosystems and assessing impacts from pollution, fishing pressure, and climate change. Data from fisheries acoustics surveys address management needs by providing a measure of biomass in management areas, detecting spatial and temporal responses in distribution relative to natural and anthropogenic impacts, and identifying hotspots that support high fish abundance or fish aggregations. Fisheries acoustics surveys conducted alongside multibeam mapping efforts inherently couple water column data with information on benthic habitats and provide information on the heterogeneity of both benthic habitats and biota in the water column. Building on this information serves to inform resource managers regarding how fishes are organized around habitat structure and the scale at which these relationships are important. Where resource managers require place-based assessments regarding the location of critical habitats along with high abundances of fish, concurrent multibeam and fisheries acoustics surveys serve as an important tool for characterizing and prioritizing coral reef ecosystems. This report summarizes the evolution of fisheries acoustics surveys onboard the NOAA ship Nancy Foster from 2008 to 2010, in conjunction with multibeam data collection, aimed at characterizing benthic and mid-water habitats in high priority conservation areas around the USVI and Puerto Rico. It also serves as a resource for the continued development of consistent data products derived from acoustic surveys. By focusing on the activities of 2010, this report highlights the progress made to date and illustrates the potential application of fisheries data derived from acoustic surveys to the management of coral reef ecosystems.

Estimation of bycatch in shrimp trawl fisheries: a comparison of estimation methods using field data and simulated data

Bycatch, or the incidental catch of nontarget organisms during fi shing operations, is a major issue in U.S. shrimp trawl fisheries. Because bycatch is typically discarded at sea, total bycatch is usually estimated by extrapolating from an observed bycatch sample to the entire fleet with either mean-per-unit or ratio estimators. Using both field observations of commercial shrimp trawlers and computer simulations, I compared five methods for generating bycatch estimates that were used in past studies, a mean-per-unit estimator and four forms of the ratio estimator, respectively: 1) the mean fish catch per unit of effort, where unit effort was a proxy for sample size, 2) the mean of the individual fish to shrimp ratios, 3) the ratio of mean fish catch to mean shrimp catch, 4) the mean of the ratios of fish catch per time fished (a variable measure of effort), and 5) the ratio of mean fish catch per mean time fished. For field data, different methods used to estimate bycatch of Atlantic croaker, spot, and weakfish yielded extremely different results, with no discernible pattern in the estimates by method, geographic region, or species. Simulated fishing fleets were used to compare bycatch estimated by the fi ve methods with “actual” (simulated) bycatch. Simulations were conducted by using both normal and delta lognormal distributions of fish and shrimp and employed a range of values for several parameters, including mean catches of fish and shrimp, variability in the catches of fish and shrimp, variability in fishing effort, number of observations, and correlations between fish and shrimp catches. Results indicated that only the mean per unit estimators provided statistically unbiased estimates, while all other methods overestimated bycatch. The mean of the individual fish to shrimp ratios, the method used in the South Atlantic Bight before the 1990s, gave the most biased estimates. Because of the statistically significant two- and 3-way interactions among parameters, it is unlikely that estimates generated by one method can be converted or corrected to estimates made by another method: therefore bycatch estimates obtained with different methods should not be compared directly.

Fecundity and spawning season of striped mullet (Mugil cephalus L.) in South Carolina estuaries

Fecundity in striped mullet (Mugil cephalus) from South Carolina correlated highly with length and weight, but not with age. Oocyte counts ranged from 4.47 × 105 to 2.52 × 106 in 1998 for fish ranging in size from 331 mm to 600 mm total length, 2.13 × 105to 3.89 × 106in 1999 for fish ranging in size from 332 mm to 588 mm total length, and 3.89 × 105 to 3.01 × 106 in 2000 for fish ranging in size from 325 mm to 592 mm total length. The striped mullet in this study had a high degree of variability in the size-at-age relation-ship; this variability was indicative of varied growth rates and compounded the errors in estimating fecundity at age. The stronger relationship of fecundity to fish size allowed a much better predictive model for potential fecundity in striped mullet. By comparing fecundity with other measures of reproductive activity, such as the gonadosomatic index, histological examination, and the measurement of mean oocyte diameters, we determined that none of these methods by themselves were adequate to determine the extent of reproductive development. Histological examinations and oocyte diameter measurements revealed that fecundity counts could be made once developing oocytes reached 0.400 μm or larger. Striped mullet are isochronal spawners; therefore fecundity estimates for this species are easier to determine because oocytes develop at approximately the same rate upon reaching 400 μm. This uniform development made oocytes that were to be spawned easier to count. When fecundity counts were used in conjunction with histological examination, oocyte diameter measurements, and gonadosomatic index, a more complete measure of reproductive potential and the timing of the spawning season was possible. In addition, it was determined that striped mullet that recruit into South Carolina estuaries spawn from October through April.

The use of agreement measures and latent class models to assess the reliability of classifying thermally marked otoliths

Otolith thermal marking is an efficient method for mass marking hatchery-reared salmon and can be used to estimate the proportion of hatchery fish captured in a mixed-stock fishery. Accuracy of the thermal pattern classification depends on the prominence of the pattern, the methods used to prepare and view the patterns, and the training and experience of the personnel who determine the presence or absence of a particular pattern. Estimating accuracy rates is problematic when no secondary marking is available and no error-free standards exist. Agreement measures, such as kappa (κ), provide a relative measure of the reliability of the determinations when independent readings by two readers are available, but the magnitude of κ can be influenced by the proportion of marked fish. If a third reader is used or if two or more groups of paired readings are examined, latent class models can provide estimates of the error rates of each reader. Applications of κ and latent class models are illustrated by a program providing contribution estimates of hatchery-reared chum and sockeye salmon in Southeast Alaska.