Distribution of catch-per-unit-of-effort and fishing effort for tuna in the Eastern Tropical Pacific Ocean by months of the year 1951-1960

ENGLISH: Comparison of physical and biological environmental factors affecting the aggregation of tunas with the success of fishing by the commercial fleets, requires that catch and effort data be examined in greater detail than has been presented in these publications. Consequently, the United States Bureau of Commercial Fisheries Biological Laboratory, San Diego, to serve the needs of its program of research on causes of variations in tuna abundance, made arrangements with the Tuna Commission to summarize these catch and effort data by month, by one-degree area, by fishing vessel size-class, for the years 1951-1960 for bait boats and 1953-1960 for purse-seiners. The present paper describes the techniques employed in summarizing these data by automatic data processing methods. It also presents the catch and effort information by months, by five-degree areas and certain combinations of five-degree areas for use by fishermen, industry personnel, and research agencies. Because of space limitations and other considerations, the one-degree tabulations are not included but are available at the Tuna Commission and Bureau laboratories. SPANISH: La comparación de los factores ambientales físicos y biológicos que afectan la agrupación del atún, con el éxito obtenido en la pesca por las flotas comerciales, requiere que los datos sobre la captura y el esfuerzo sean examinados con mayor detalle de lo que han sido presentados en estas publicaciones. En consecuencia, el Laboratorio Biológico del Buró de Pesquerías Comerciales de los Estados Unidos, situado en San Diego, a fin de llenar los requisitos de su programa de investigación sobre las causas de las variaciones en la abundancia del atún, hizo arreglos con la Comisión del Atún para sumarizar esos datos sobre la captura y el esfuerzo por meses, por áreas de un grado, por clases de tamaño de las embarcaciones de pesca durante los años 1951-1960 en lo que concierne a los barcos de carnada y durante el período 1953-1960 en lo que respecta a los barcos rederos. El presente trabajo describe la técnica empleada en la sumarización de dichos datos mediante métodos automáticos de manejo de datos. También se da aquí la información sobre la captura y el esfuerzo por meses, por áreas de cinco grados y ciertas combinaciones de áreas de cinco grados para el uso de los pescadores, del personal de la industria y de las oficinas de investigación. Por falta de espacio y otras razones, las tabulaciones de las áreas de un grado no han sido incluídos en este trabajo, pero están a la disposición de quien tenga interés en los laboratorios de la Comisión del Atún y del Buró.

Framework for the Shape Optimization of Aerodynamic Profiles Using Genetic Algorithms

This study developed a framework for the shape optimization of aerodynamics profiles using computational fluid dynamics (CFD) and genetic algorithms. Agenetic algorithm code and a commercial CFD code were integrated to develop a CFD shape optimization tool. The results obtained demonstrated the effectiveness of the developed tool. The shape optimization of airfoils was studied using different strategies to demonstrate the capacity of this tool with different GA parameter combinations.

Auswirkungen der Wassertemperatur auf die Missbildungshäufigkeit pelagischer Fischembryonen

Malformation rates in embryos of dab, whiting, cod, flounder and plaice have been monitored for several years (1984-2006) in the Southern North Sea. For embryos of all species investigated trends for the fluctuation of malformation rates over the time were registered in the areas showing intermediate prevalences at the beginning of the studies in 1984 and maxima in 1987. Thereafter for all species a decrease of malformation rates was found until 2006 excepting an increase in 1996. A significant negative correlation existed between surface water temperature and prevalences of malformed embryos of dab and other species.

Cost effectiveness of a combination of instruments for global warming: a quantitative approach for Spain

Climate change is an important environmental problem and one whose economic implications are many and varied. This paper starts with the presumption that mitigation of greenhouse gases is a necessary policy that has to be designed in a cost effective way. It is well known that market instruments are the best option for cost effectiveness. But the discussion regarding which of the various market instruments should be used, how they may interact and what combinations of policies should be implemented is still open and very lively. In this paper we propose a combination of instruments: the marketable emission permits already in place in Europe for major economic sectors and a CO(2) tax for economic sectors not included in the emissions permit scheme. The study uses an applied general equilibrium model for the Spanish economy to compute the results obtained with the new mix of instruments proposed. As the combination of the market for emission permits and the CO(2) tax admits different possibilities that depend on how the mitigation is distributed among the economic sectors, we concentrate on four possibilities: cost-effective, equalitarian, proportional to emissions, and proportional to output distributions. Other alternatives to the CO(2) tax are also analysed (tax on energy, on oil and on electricity). Our findings suggest that careful, well designed policies are needed as any deviation imposes significant additional costs that increase more than proportionally to the level of emissions reduction targeted by the EU.

Design and synthesis of quasi-diastereomeric molecules with unchanging central, regenerating axial and switchable helical chirality via cleavage and formation of Ni(II)–O and Ni(II)–N coordination bonds

9 p.

Materials and methods for tagging purse seine and baitboat caught tunas

ENGLISH: The percentages of return of purse seine-tagged yellowfin and baitboat- tagged yellowfin and skipjack subjected to various combinations of treatments were tested statistically to determine the effects of the treatments. For yellowfin the precentages of return differed among sets or dates of release, and were higher for double-tagged fish, larger fish, and fish released soonest during each set. For skipjack the percentages of return differed among dates of release, and were higher for single-tagged fish and fish tagged in plastic-covered cradles. SPANISH: Los porcentajes de retorno de aleta amarilla marcado en barcos cerqueros y los de aleta amarilla y barrilete marcados en los de carnada sometidos a varias combinaciones de tratamientos fueron probados estadísticamente para determinar los efectos de los tratamientos. Con respecto al aleta amarilla los porcentajes de retorno fueron diferentes entre los lances o fechas de liberación, y fueron más altos para los peces con doble marca, peces de más talla y peces liberados lo más pronto durante cada lance. Los porcentajes de retorno del barrilete fueron diferentes entre las fechas de liberación, y fueron más altos para peces con una sola marca y peces marcados en cunas cubiertas con plástico. (PDF contains 41 pages.)

Zusammenhang zwischen Schadstoffen aus der Elbe und missgebildeten Fischembryonen in der Deutschen Bucht.

The trends of malformation prevalence in embryos of dab, Limanda limanda, in the southern North Sea after the year 1990 mirrored the drop in major pollutants in the rivers draining into the German Bight. Despite this general decline we detected a pollution event in the southern North Sea in winter 1995/1996 employing the prevalence of malformations in dab embryos as an indicator. An abrupt rise in malformation prevalence in the embryos of dab, corresponded to a dramatic increase in DDT levels in parent fish from the same area, indicating a hitherto unnoticed introduction of considerable quantities of DDT into the system. This input could be traced back to discharges of unknown origen into the River Elbe.

Mateda-2.0: Estimation of Distribution Algorithms in MATLAB

This paper describes Mateda-2.0, a MATLAB package for estimation of distribution algorithms (EDAs). This package can be used to solve single and multi-objective discrete and continuous optimization problems using EDAs based on undirected and directed probabilistic graphical models. The implementation contains several methods commonly employed by EDAs. It is also conceived as an open package to allow users to incorporate different combinations of selection, learning, sampling, and local search procedures. Additionally, it includes methods to extract, process and visualize the structures learned by the probabilistic models. This way, it can unveil previously unknown information about the optimization problem domain. Mateda-2.0 also incorporates a module for creating and validating function models based on the probabilistic models learned by EDAs.

Prioritization of candidate cancer genes-an aid to oncogenomic studies

The development of techniques for oncogenomic analyses such as array comparative genomic hybridization, messenger RNA expression arrays and mutational screens have come to the fore in modern cancer research. Studies utilizing these techniques are able to highlight panels of genes that are altered in cancer. However, these candidate cancer genes must then be scrutinized to reveal whether they contribute to oncogenesis or are coincidental and non-causative. We present a computational method for the prioritization of candidate (i) proto-oncogenes and (ii) tumour suppressor genes from oncogenomic experiments. We constructed computational classifiers using different combinations of sequence and functional data including sequence conservation, protein domains and interactions, and regulatory data. We found that these classifiers are able to distinguish between known cancer genes and other human genes. Furthermore, the classifiers also discriminate candidate cancer genes from a recent mutational screen from other human genes. We provide a web-based facility through which cancer biologists may access our results and we propose computational cancer gene classification as a useful method of prioritizing candidate cancer genes identified in oncogenomic studies.

Änderung wissenschaftlicher Gattungsnamen der Echten Rochen (Raja spp.), Familie Rajidae

Based on a comprehensive compilation and evaluation of hitherto published revisions by international rajid systematists, the U.S. authors McEachran and Dunn (1998) published a summarizing revision on phylogenetic interrelationships within the family Rajidae. The family has been subdivided into the two subfamilies Rajinae and Arhynchobatinae comprising 15 and 11 rajid genera, respectively. Within the Rajinae, seven former subgenera of Raja and Gurgesiella have been newly elevated to generic rank, likewise two former subgenera of Raja within the Arhynchobatinae. As a consequence, generic names have changed for quite a number of mainly former Raja species not only in European and North Atlantic waters. Table 1 presents for comparison a selection of European skate species on the shelf with the former and new generic assignment, as well as their German names. As Fishery Conventions and Societies, as well as regional (e.g., ICES) and international organizations (e.g., FAO) are increasingly using the new scientific name combinations for fishery statistics and in relevant publications, all involved in fisheries should be aware of these nomenclatorial alterations.

Estimation of the abundance of yellowfin tuna, Thunnus albacares, by age groups and regions within the Eastern Pacific Ocean

ENGLISH: Monthly estimates of the abundance of yellowfin tuna by age groups and regions within the eastern Pacific Ocean during 1970-1988 are made, using purse-seine catch rates, length-frequency samples, and results from cohort analysis. The numbers of individuals caught of each age group in each logged purse-seine set are estimated, using the tonnage from that set and length-frequency distribution from the "nearest" length-frequency sample(s). Nearest refers to the closest length frequency sample(s) to the purse-seine set in time, distance, and set type (dolphin associated, floating object associated, skipjack associated, none of these, and some combinations). Catch rates are initially calculated as the estimated number of individuals of the age group caught per hour of searching. Then, to remove the effects of set type and vessel speed, they are standardized, using separate weiznted generalized linear models for each age group. The standardized catch rates at the center of each 2.5 0 quadrangle-month are estimated, using locally-weighted least-squares regressions on latitude, longitude and date, and then combined into larger regions. Catch rates within these regions are converted to numbers of yellowfin, using the mean age composition from cohort analysis. The variances of the abundance estimates within regions are large for 0-, 1-, and 5-year-olds, but small for 1.5- to 4-year-olds, except during periods of low fishing activity. Mean annual catch rate estimates for the entire eastern Pacific Ocean are significantly positively correlated with mean abundance estimates from cohort analysis for age groups ranging from 1.5 to 4 years old. Catch-rate indices of abundance by age are expected to be useful in conjunction with data on reproductive biology to estimate total egg production within regions. The estimates may also be useful in understanding geographic and temporal variations in age-specific availability to purse seiners, as well as age-specific movements. SPANISH: Se calculan estimaciones mensuales de la abundancia del atún aleta amarilla por grupos de edad y regiones en el Océano Pacífico oriental durante 1970-1988, usando tasas de captura cerquera, muestras de frecuencia de talla, y los resultados del análisis de cohortes. Se estima el número de individuos capturados de cada grupo de edad en cada lance cerquero registrado, usando el tonelaje del lance en cuestión y la distribución de frecuencia de talla de la(s) muestra(s) de frecuencia de talla "más cercana/s)," "Más cercana" significa la(s) muestra(s) de frecuencia de talla más parecida(s) al lance cerquero en cuanto a fecha, distancia, y tipo de lance (asociado con delfines, con objeto flotante, con barrilete, con ninguno de éstos, y algunas combinaciones). Se calculan inicialmente las tasas de captura como el número estimado de individuos del grupo de edad capturado por hora de búsqueda. A continuación, para eliminar los efectos del tipo de lance y la velocidad del barco, se estandardizan dichas tasas, usando un modelo lineal generalizado ponderado, para cada grupo por separado. Se estima la tasa de captura estandardizada al centro de cada cuadrángulo de 2.5°-mes, usando regresiones de mínimos cuadrados ponderados localmente por latitud, longitud, y fecha, y entonces combinándolas en regiones mayores. Se convierten las tasas de captura dentro de estas regiones en números de aletas amarillas individuales, usando el número promedio por edad proveniente del análisis de cohortes. Las varianzas de las estimaciones de la abundancia dentro de las regiones son grandes para los peces de O, 1, Y5 años de edad, pero pequeñas para aquellos de entre 1.5 Y4 años de edad, excepto durante períodos de poca actividad pesquera. Las estimaciones de la tasa de captura media anual para todo el Océano Pacífico oriental están correlacionadas positivamente de forma significativa con las estimaciones de la abundancia media del análisis de las cohortes para los grupos de edad de entre 1.5 y 4 años. Se espera que los índices de abundancia por edad basados en las tasas de captura sean útiles, en conjunto con datos de la biología reproductiva, para estimar la producción total de huevos por regiones. Las estimaciones podrían asimismo ser útiles para la comprensión de las variaciones geográficas y temporales de la disponibilidad específica por edad a los barcos cerqueros, y también las migraciones específicas por edad. (PDF contains 35 pages.)

An evaluation of length-frequency sampling procedures and subsequent data analysis for purse seine-caught yellowfin tuna in the Eastern Pacific Ocean

ENGLISH: A two-stage sampling design is used to estimate the variances of the numbers of yellowfin in different age groups caught in the eastern Pacific Ocean. For purse seiners, the primary sampling unit (n) is a brine well containing fish from a month-area stratum; the number of fish lengths (m) measured from each well are the secondary units. The fish cannot be selected at random from the wells because of practical limitations. The effects of different sampling methods and other factors on the reliability and precision of statistics derived from the length-frequency data were therefore examined. Modifications are recommended where necessary. Lengths of fish measured during the unloading of six test wells revealed two forms of inherent size stratification: 1) short-term disruptions of existing pattern of sizes, and 2) transition zones between long-term trends in sizes. To some degree, all wells exhibited cyclic changes in mean size and variance during unloading. In half of the wells, it was observed that size selection by the unloaders induced a change in mean size. As a result of stratification, the sequence of sizes removed from all wells was non-random, regardless of whether a well contained fish from a single set or from more than one set. The number of modal sizes in a well was not related to the number of sets. In an additional well composed of fish from several sets, an experiment on vertical mixing indicated that a representative sample of the contents may be restricted to the bottom half of the well. The contents of the test wells were used to generate 25 simulated wells and to compare the results of three sampling methods applied to them. The methods were: (1) random sampling (also used as a standard), (2) protracted sampling, in which the selection process was extended over a large portion of a well, and (3) measuring fish consecutively during removal from the well. Repeated sampling by each method and different combinations indicated that, because the principal source of size variation occurred among primary units, increasing n was the most effective way to reduce the variance estimates of both the age-group sizes and the total number of fish in the landings. Protracted sampling largely circumvented the effects of size stratification, and its performance was essentially comparable to that of random sampling. Sampling by this method is recommended. Consecutive-fish sampling produced more biased estimates with greater variances. Analysis of the 1988 length-frequency samples indicated that, for age groups that appear most frequently in the catch, a minimum sampling frequency of one primary unit in six for each month-area stratum would reduce the coefficients of variation (CV) of their size estimates to approximately 10 percent or less. Additional stratification of samples by set type, rather than month-area alone, further reduced the CV's of scarce age groups, such as the recruits, and potentially improved their accuracy. The CV's of recruitment estimates for completely-fished cohorts during the 198184 period were in the vicinity of 3 to 8 percent. Recruitment estimates and their variances were also relatively insensitive to changes in the individual quarterly catches and variances, respectively, of which they were composed. SPANISH: Se usa un diseño de muestreo de dos etapas para estimar las varianzas de los números de aletas amari11as en distintos grupos de edad capturados en el Océano Pacifico oriental. Para barcos cerqueros, la unidad primaria de muestreo (n) es una bodega de salmuera que contenía peces de un estrato de mes-área; el numero de ta11as de peces (m) medidas de cada bodega es la unidad secundaria. Limitaciones de carácter practico impiden la selección aleatoria de peces de las bodegas. Por 10 tanto, fueron examinados los efectos de distintos métodos de muestreo y otros factores sobre la confiabilidad y precisión de las estadísticas derivadas de los datos de frecuencia de ta11a. Se recomiendan modificaciones donde sean necesarias. Las ta11as de peces medidas durante la descarga de seis bodegas de prueba revelaron dos formas de estratificación inherente por ta11a: 1) perturbaciones a corto plazo en la pauta de ta11as existente, y 2) zonas de transición entre las tendencias a largo plazo en las ta11as. En cierto grado, todas las bodegas mostraron cambios cíclicos en ta11a media y varianza durante la descarga. En la mitad de las bodegas, se observo que selección por ta11a por los descargadores indujo un cambio en la ta11a media. Como resultado de la estratificación, la secuencia de ta11as sacadas de todas las bodegas no fue aleatoria, sin considerar si una bodega contenía peces de un solo lance 0 de mas de uno. El numero de ta11as modales en una bodega no estaba relacionado al numero de lances. En una bodega adicional compuesta de peces de varios lances, un experimento de mezcla vertical indico que una muestra representativa del contenido podría estar limitada a la mitad inferior de la bodega. Se uso el contenido de las bodegas de prueba para generar 25 bodegas simuladas y comparar los resultados de tres métodos de muestreo aplicados a estas. Los métodos fueron: (1) muestreo aleatorio (usado también como norma), (2) muestreo extendido, en el cual el proceso de selección fue extendido sobre una porción grande de una bodega, y (3) medición consecutiva de peces durante la descarga de la bodega. EI muestreo repetido con cada método y distintas combinaciones de n y m indico que, puesto que la fuente principal de variación de ta11a ocurría entre las unidades primarias, aumentar n fue la manera mas eficaz de reducir las estimaciones de la varianza de las ta11as de los grupos de edad y el numero total de peces en los desembarcos. El muestreo extendido evito mayormente los efectos de la estratificación por ta11a, y su desempeño fue esencialmente comparable a aquel del muestreo aleatorio. Se recomienda muestrear con este método. El muestreo de peces consecutivos produjo estimaciones mas sesgadas con mayores varianzas. Un análisis de las muestras de frecuencia de ta11a de 1988 indico que, para los grupos de edad que aparecen con mayor frecuencia en la captura, una frecuencia de muestreo minima de una unidad primaria de cada seis para cada estrato de mes-área reduciría los coeficientes de variación (CV) de las estimaciones de ta11a correspondientes a aproximadamente 10% 0 menos. Una estratificación adicional de las muestras por tipo de lance, y no solamente mes-área, redujo aun mas los CV de los grupos de edad escasos, tales como los reclutas, y mejoró potencialmente su precisión. Los CV de las estimaciones del reclutamiento para las cohortes completamente pescadas durante 1981-1984 fueron alrededor de 3-8%. Las estimaciones del reclutamiento y sus varianzas fueron también relativamente insensibles a cambios en las capturas de trimestres individuales y las varianzas, respectivamente, de las cuales fueron derivadas. (PDF contains 70 pages)

Chromosomal and histological evidences of infertility in F2 and F2 backcross. Hybrid generations of Clarias anguillaris and Heterobranchus longifilis

Male meiosis was studied in 9 different mating combinations in parental, first, second and backcross generation hybrids of Clarias anguillaris and Heterobranchus longifilis. 27 bivalents were recorded in metaphase I for seven mating combinations. The number of bivalents in F1 hybrid male x C. anguillaris female could not be determined due to a high degree of clumping of the chromosomes. All metaphase I cells observed in female F1 hybrid x male H. longifilis had three complex bivalents consisting of 43.3% giant ring and 56.7% giant rod chromosomes. The number of ring bivalents per cell was higher in parental H. longifilis than parental C. anguillaris. The number of ring bivalents per cell increased from F1 (6.7 and 8.2) to F2 backcross (13.5) hybrid generations indicating increasing chromosomal instability of backcross hybrids over Fl and F2 hybrids

Negative regulators of a growth factor-mediated signaling pathway in the nematode Caenorhabditis elegans

Vulval differentiation in C. elegans is mediated by an Epidermal growth factor (EGF)- EGF receptor (EGFR) signaling pathway. I have cloned unc-101, a negative regulator of vulval differentiation of the nematode C. elegans. unc-101 encodes a homolog of AP47, the medium chain of the trans-Golgi clathrin-associated protein complex. This identity was confirmed by cloning and comparing sequence of a C. elegans homolog of AP50, the medium chain of the plasma membrane clathrin-associated protein complex. I provided the first genetic evidence that the trans-Golgi clathrin-coated vesicles are involved in regulation of an EGF signaling pathway. Most of the unc-101 alleles are deletions or nonsense mutations, suggesting that these alleles severely reduce the unc-101 activity. A hybrid gene that contains parts of unc-101 and mouse AP4 7 rescued at least two phenotypes of unc-101 mutations, the Unc and the suppression of vulvaless phenotype of let-23(sy1) mutation. Therefore, the functions of AP47 are conserved between nematodes and mammals.

unc-101 mutations can cause a greater than wild-type vulval differentiation in combination with certain mutations in sli-1, another negative regulator of the vulval induction pathway. A mutation in a new gene, rok-1, causes no defect by itself, but causes a greater than wild-type vulval differentiation in the presence of a sli-1 mutation. The unc-101; rok-1; sli-1 triple mutants display a greater extent of vulval differentiation than any double mutant combinations of unc-101, rok-1 and sli-1. Therefore, rok-1 locus defines another negative regulator of the vulval induction pathway.

I analyzed a second gene encoding an AP47 homolog in C. elegans. This gene, CEAP47, encodes a protein 72% identical to both unc-101 and mammalian AP47. A hybrid gene containing parts of unc-101 and CEAP47 sequences can rescue phenotypes of unc-101 mutants, indicating that UNC- 101 and CEAP47 proteins can be redundant if expressed in the same set of cells.

Structural requirements for catalysis: studies on RTEM-1 β-lactamase

β-lactamases are a group of enzymes that confer resistance to penam and cephem antibiotics by hydrolysis of the β-lactam ring, thereby inactivating the antibiotic. Crystallographic and computer modeling studies of RTEM-1 β-lactamase have indicated that Asp 132, a strictly conserved residue among the class A β-lactamases, appears to be involved in substrate binding, catalysis, or both. To study the contribution of residue 132 to β-lactamase function, site saturation mutagenesis was used to generate mutants coding for all 20 amino acids at position 132. Phenotypic screening of all mutants indicated that position 132 is very sensitive to amino acid changes, with only N132C, N132D, N132E, and N132Q showing any appreciable activity. Kinetic analysis of three of these mutants showed increases in K_M, along with substantial decreases in k_(cat). Efforts to trap a stable acyl-enzyme intermediate were unsuccessfuL These results indicate that residue 132 is involved in substrate binding, as well as catalysis, and supports the involvement of this residue in acylation as suggested by Strynadka et al.

Crystallographic and computer modeling studies of RTEM-1 β-lactamase have indicated that Lys 73 and Glu 166, two strictly conserved residues among the class A β-lactamases, appear to be involved in substrate binding, catalysis, or both. To study the contribution of these residues to β-lactamase function, site saturation mutagenesis was used to generate mutants coding for all 20 amino acids at positions 73 and 166. Then all 400 possible combinations of mutants were created by combinatorial mutagenesis. The colonies harboring the mutants were screened for growth in the presence of ampicillin. The competent colonys' DNA were sequenced, and kinetic parameters investigated. It was found that lysine is essential at position 73, and that position 166 only tolerated fairly conservative changes (Aspartic acid, Histidine, and Tyrosine). These functional mutants exhibited decreased kcat's, but K_M was close to wild-type levels. The results of the combinatorial mutagenesis experiments indicate that Lysis absolutely required for activity at position 73; no mutation at residue 166 can compensate for loss of the long side chain amine. The active mutants found--K73K/E166D, K73KIE166H, and K73KIE166Y were studied by kinetic analysis. These results reaffirmed the function of residue 166 as important in catalysis, specifically deacylation.

The identity of the residue responsible for enhancing the active site serine (Ser 70) in RTEM-1 β-lactamase has been disputed for some time. Recently, analysis of a crystal structure of RTEM-1 β-lactamase with covalently bound intermediate was published, and it was suggested that Lys 73, a strictly conserved residue among the class A β-lactamases, was acting as a general base, activating Ser 70. For this to be possible, the pK_a of Lys 73 would have to be depressed significantly. In an attempt to assay the pK_a of Lys 73, the mutation K73C was made. This mutant protein can be reacted with 2-bromoethylamine, and activity is restored to near wild type levels. ^(15)N-2-bromoethylamine hydrobromide and ^(13)C-2-bromoethylamine hydrobromide were synthesized. Reacting these compounds with the K73C mutant gives stable isotopic enrichment at residue 73 in the form of aminoethylcysteine, a lysine homologue. The pK_a of an amine can be determined by NMR titration, following the change in chemical shift of either the ^(15)N-amine nuclei or adjacent Be nuclei as pH is changed. Unfortunately, low protein solubility, along with probable label scrambling in the Be experiment, did not permit direct observation of either the ^(15)N or ^(13)C signals. Indirect detection experiments were used to observe the protons bonded directly to the ^(13)C atoms. Two NMR signals were seen, and their chemical shift change with pH variation was noted. The peak which was determined to correspond to the aminoethylcysteine residue shifted from 3.2 ppm down to 2.8 ppm over a pH range of 6.6 to 12.5. The pK_a of the amine at position 73 was determined to be ~10. This indicates that residue 73 does not function as a general base in the acylation step of the reaction. However the experimental measurement takes place in the absence of substrate. Since the enzyme undergoes conformational changes upon substrate binding, the measured pK_a of the free enzyme may not correspond to the pK_a of the enzyme substrate complex.