Modelos multivariados en la búsqueda de regiones genómicas para resistencia a mal de río cuarto y bacteriosis en maíz / Multi-trait models for genomic regions associated with mal de río cuarto and bacterial disease in maize

RESUMEN La producción de maíz (Zea Mays L.) ha sido ampliamente beneficiada con la mejora de líneas endocriadas respecto a la resistencia a enfermedades causadas por virus y hongos. Sin embargo, es notable la ausencia de genotipos resistentes a bacteriosis. El objetivo del presente estudio fue identificar regiones genómicas para la mejora de resistencia a Mal de Río Cuarto (MRC) y a bacteriosis (BD) en un germoplasma diverso de maíz. Se evaluó, para ambas enfermedades, una población diversa de líneas de maíz en el ciclo de cultivo 2019-2020 en la región argentina donde la virosis MRC es endémica. Se estimó incidencia y severidad de MRC y BD en cada línea y se realizó un estudio de mapeo por asociación (GWAS) con 78.376 marcadores SNPs. Un modelo multicarácter se utilizó para evaluar simultáneamente la resistencia a MRC y BD en las líneas evaluadas. El germoplasma evidenció alta variabilidad genética tanto para la mejora de la resistencia a MRC como a BD, pero no se observó correlación genética significativa entre la respuesta a ambas enfermedades. Se identificaron regiones genómicas promisorias para resistencia a MRC y a BD, que serán confirmadas en evaluaciones en nuevos ambientes.

ABSTRACT Maize (Zea Mays L.) production has been greatly benefited from the improvement of inbred lines in regard to the resistance to diseases. However, the absence of resistant genotypes to bacteriosis is remarkable. The aim of the study was to identify genomic regions for resistance to Mal de Río Cuarto (MRC) and to bacterial disease (BD) in a diverse maize germplasm evaluated in the Argentinian region where MRC virus is endemic. A maize diverse population was assessed for both diseases during the 2019-2020 crop season. Incidence and severity of MRC and BD were estimated for each line and a genome wide association study (GWAS) was conducted with 78,376 SNP markers. A multi-trait mixed linear model was used for simultaneous evaluation of resistance to MRC and BD in the scored lines. The germplasm showed high genetic variability for both MRC and BD resistance. No significant genetic correlation was observed between the response to both diseases. Promising genomic regions for resistance to MRC and BD were identified and will be confirmed in further trials.

Alelos asociados al índice de severidad de la enfermedad Mal de Río Cuarto en germoplasma exótico de maíz / Alleles associated to disease severity index of Mal de Río Cuarto disease in maize exotic germplasm

RESUMEN El Mal de Río Cuarto (MRC) es una de las enfermedades virales más importantes del maíz en Argentina. El índice de severidad de enfermedad (ISE) permite combinar la incidencia y la severidad de una enfermedad en una métrica única. La reacción genotípica a MRC ha sido muy estudiada en poblaciones biparentales, sin embargo este carácter complejo no se ha analizado mediante estudios de mapeo por asociación. El objetivo del presente trabajo es identificar nuevos alelos de resistencia asociados con el ISE de la enfermedad MRC de maíz en un germoplasma exótico del Centro Internacional de Mejoramiento de Maíz y Trigo (CIMMYT). Una población de líneas de maíz del CIMMYT se evaluó fenotípicamente en ambientes donde la enfermedad es endémica. Los predictores del efecto genotípico (BLUP, best linear unbiased predictor) del ISE de MRC y 78.376 marcadores SNP (Single Nucleotide Polymorphism) se usaron para realizar el mapeo por asociación en 186 líneas de maíz. Los componentes de varianza y los valores de heredabilidad sugieren una amplia variabilidad genotípica en la población de líneas. El mapeo por asociación permitió identificar 11 posibles QTL de resistencia a MRC. La incorporación de germoplasma exótico en los programas de mejoramiento de maíz locales podría contribuir favorablemente a la creación de genotipos híbridos con mayor nivel de resistencia a MRC. La capacidad predictiva de los marcadores asociados con la resistencia a MRC indican que la selección asistida por marcadores es una herramienta recomendable para seleccionar genotipos resistentes a MRC.

ABSTRACT Mal de Río Cuarto (MRC) is one of the most important viral diseases of maize in Argentina. The disease severity index (DSI) allows to combine the incidence and severity of a disease in a single metric. The genotypic reaction to MRC has been extensively studied in biparental populations. However, this complex trait has not been analyzed by genome-wide association studies (GWAS). The aim of this work is to identify new resistance alleles associated with DSI of MRC in an exotic germplasm from the International Maize and Wheat Improvement Center (CIMMYT). A population of maize lines from CIMMYT was phenotypically evaluated in environments in the area where the disease is endemic. The predictors of genetic effects (BLUP, best linear unbiased predictor) and 78,376 SNP markers (Single Nucleotide Polymorphism) were used to perform the GWAS in 186 maize lines. The values of variance components and mean-basis heritability suggest a wide genotypic variability in the population. The GWAS allowed to identify 11 putative QTL of resistance to MRC. The incorporation of exotic germplasm into local maize breeding programs could contribute favorably to the creation of hybrids with a higher level of resistance to MRC. The predictive ability of associated markers with MRC resistance indicates that marker-assisted selection is an advisable tool for selecting MRC resistant genotypes.

Diversidad genotípica de 291 líneas de maíz de CIMMYT y caracterización fenotípica en el sur de Córdoba, Argentina / Genotypic diversity in 291 maize lines from CIMMYT and phenotypic characterization in southern Córdoba, Argentina

El maíz (Zea mays L.) posee un genoma complejo y una amplia diversidad genética. La información de caracteres fenotípicos y marcadores moleculares en su conjunto provee una mejor descripción e interpretación de la variabilidad genética. El objetivo del trabajo fue estimar la diversidad genética y caracterizar fenotípicamente un panel de 291 líneas de maíz de CIMMYT. Las líneas corresponden a ocho grupos establecidos de acuerdo a su adaptación ambiental y origen geográfico. Éstas se evaluaron fenotípicamente por medio de nueve caracteres agro-morfológicos en tres ambientes del sur de la provincia de Córdoba, Argentina. El intervalo antesis-estigma (IAE) presentó la mayor variabilidad fenotípica. El 40% de las líneas de maíz registraron un IAE menor a cinco días, indicando buena adaptación a los ambientes de evaluación. La estructura poblacional dada por los subgrupos de adaptación ambiental es sólo un factor menor que contribuye a la variabilidad fenotípica del panel estudiado. El análisis de componentes principales (ACP) permitió obtener el ordenamiento fenotípico y el genotípico, mientras que el análisis de procrustes generalizado indicó un consenso del 60% entre ambos ordenamientos para el total de líneas. El consenso entre el ordenamiento obtenido con caracteres agro-morfológicos y con marcadores moleculares indica desequilibrio de ligamiento entre los SNPs y los genes que controlan los caracteres agro-morfológicos. Los resultados muestran una amplia diversidad genética en el germoplasma evaluado, lo que sugiere que esta colección de líneas es un recurso importante para impulsar ganancias genéticas futuras en los programas de mejoramiento de maíz de Argentina.

CIMMYT maize inbred lines (CMLs) are freely distributed to breeding programs around the world. Better information on phenotypic and genotypic diversity may provide guidance to breeders on how to use more efficiently the CMLs in their breeding programs. In this study a group of 291 CIMMYT maize inbred lines, was phenotyped by nine agro-morphological traits in south Córdoba, Argentina and genotyped using 18,082 SNPs. Based on the geographic information and the environmental adaptation, 291 CMLs were classified into eight subgroups. Anthesis-silking interval (IAE) was the trait with higher phenotypic diversity. A 40% of maize inbred lines, with IAE less than five days, show a good adaptation to growing conditions in south Córdoba, Argentina. The low phenotypic variation explained by environmental adaptation subgroups indicates that population structure is only a minor factor contributing to phenotypic diversity in this panel. Principal component analysis (ACP) allowed us to obtain phenotypic and genotypic orderings. Generalized procrustes analysis (APG) indicated a 60% consensus between both data type from the total panel of maize lines. In each environmental adaptation subgroup, the APG consensus was higher. This result, which might indicate linkage disequilibrium between SNPs markers and the genes controlling these agro-morphological traits, is promising and could be used as an initial tool in the identification of Quantitative Trait Loci (QTL). Information on genetic diversity, population structure and phenotypic diversity in local environments will help maize breeders to better understand how to use the current CIMMYT maize inbred lines group.

Potentially synbiotic fermented beverage with aqueous extracts of quinoa (Chenopodium quinoa Willd) and soy.

The aim of this study was to develop a potentially synbiotic beverage fermented with Lactobacillus casei LC-1 based on aqueous extracts of soy and quinoa with added fructooligosaccharides (FOS). Five formulations with differing proportions of soy and quinoa extracts were tested. The viability of the microorganism, the pH, and the acidity of all formulations were monitored until the 28th day of storage at 5 ℃. The chemical composition of the extracts and beverages and the rheological and sensory properties of the final products were analyzed. Although an increase in acidity and a decrease in pH were observed during the 28 days of storage, the viability of the probiotic microorganism was maintained at 10(8) CFU·mL(-1) in all formulated beverages throughout the storage period. An increase in viscosity and consistency in the formulations with higher concentrations of quinoa (F1 and F2) was observed. Formulation F4 (70% soy and 30% quinoa extracts) showed the least hysteresis. Formulations F4 and F5 (100% soy extract) had the best sensory acceptance while F4 resulted in the highest intention to purchase from a group of 80 volunteers. For chemical composition, F3 (50% soy and 50% quinoa extracts) and F4 showed the best results compared to similar fermented beverages. The formulation F4 was considered the best beverage overall.

Anti-CEA antibody fragments labeled with [(18)F]AlF for PET imaging of CEA-expressing tumors.

A facile and rapid method to label peptides with (18)F based on chelation of [(18)F]AlF has been developed recently. Since this method requires heating to 100 °C, it cannot be used to label heat-sensitive proteins. Here, we used a two-step procedure to prepare (18)F-labeled heat-labile proteins using the [(18)F]AlF method based on hot maleimide conjugation. 1,4,7-Triazacyclononae-1,4-diacetate (NODA) containing a methyl phenylacetic acid group (MPA) functionalized with N-(2-aminoethyl)maleimide (EM) was used as a ligand which was labeled with [(18)F]AlF and then conjugated to the humanized anti-CEA antibody derivatives hMN-14-Fab', hMN-14-(scFv)2 (diabody), and a Dock-and-Lock engineered dimeric fragment hMN-14 Fab-AD2 at room temperature. The in vivo tumor targeting characteristics of the (18)F-labeled antibody derivatives were determined by PET imaging of mice with s.c. xenografts. NODA-MPAEM was radiolabeled with [(18)F]AlF at a specific activity of 29-39 MBq/nmol and a labeling efficiency of 94 ± 2%. The labeling efficiencies of the maleimide conjugation ranged from 70% to 77%, resulting in [(18)F]AlF-labeled hMN14-Fab', hMN14-Fab-AD2, or hMN14-diabody with a specific activity of 15-17 MBq/nmol. The radiolabeled conjugates were purified by gel filtration. For biodistribution and microPET imaging, antibody fragments were injected intravenously into BALB/c nude mice with s.c. CEA-expressing LS174T xenografts (right flank) and CEA-negative SK-RC-52 xenografts (left flank). All [(18)F]AlF-labeled conjugates showed specific uptake in the LS174T xenografts with a maximal tumor uptake of 4.73% ID/g at 4 h after injection. Uptake in CEA-negative SK-RC-52 xenografts was significantly lower. Tumors were clearly visualized on microPET images. Using a [(18)F]AlF-labeled maleimide functionalized chelator, antibody fragments could be radiofluorinated within 4 h at high specific activity. Here, we translated this method to preclinical PET imaging studies and showed feasibility of [(18)F]AlF-fluorinated hMN-14-Fab', [(18)F]AlF-hMN-14-Fab-AD2, and [(18)F]AlF-hMN-14-diabody for microPET imaging of CEA-expressing colonic cancer.

Development of an imaging-guided CEA-pretargeted radionuclide treatment of advanced colorectal cancer: first clinical results.

BACKGROUND: Radiolabelled antibody targeting of cancer is limited by slow blood clearance. Pretargeting with a non-radiolabelled bispecific monoclonal antibody (bsMAb) followed by a rapidly clearing radiolabelled hapten peptide improves tumour localisation. The primary goals of this first pretargeting study in patients with the anti-CEACAM5 × anti-hapten (HSG) bsMAb, TF2, and the radiolabelled hapten-peptide, IMP288, were to assess optimal pretargeting conditions and safety in patients with metastatic colorectal cancer (CRC). METHODS: Different dose schedules were studied in four cohorts of five patients: (1) shortening the interval between the bsMAb and peptide administration (5 days vs 1 day), (2) escalating the TF2 dose (from 75 to 150 mg), and (3) reducing the peptide dose (from 100 to 25 µg). After confirmation of tumour targeting by (111)In-IMP288, patients were treated with a bsMAb/(177)Lu-IMP288 cycle. RESULTS: Rapid and selective tumour targeting of the radiolabelled peptide was visualised within 1 h, with high tumour-to-tissue ratios (>20 at 24 h). Improved tumour targeting was achieved with a 1-day interval between the administration of the bsMAb and the peptide and with the 25-µg peptide dose. High (177)Lu-IMP288 doses (2.5-7.4 GBq) were well tolerated, with some manageable TF2 infusion reactions, and transient grades 3-4 thrombocytopaenia in 10% of the patients who received (177)Lu-IMP288. CONCLUSION: This phase I study demonstrates for the first time that pretargeting with bsMAb TF2 and radiolabelled IMP288 in patients with CEA-expressing CRC is feasible and safe. With this pretargeting method, tumours are specifically and rapidly targeted.

Imaging retinal mosaics in the living eye.

Adaptive optics imaging of cone photoreceptors has provided unique insight into the structure and function of the human visual system and has become an important tool for both basic scientists and clinicians. Recent advances in adaptive optics retinal imaging instrumentation and methodology have allowed us to expand beyond cone imaging. Multi-wavelength and fluorescence imaging methods with adaptive optics have allowed multiple retinal cell types to be imaged simultaneously. These new methods have recently revealed rod photoreceptors, retinal pigment epithelium (RPE) cells, and the smallest retinal blood vessels. Fluorescence imaging coupled with adaptive optics has been used to examine ganglion cells in living primates. Two-photon imaging combined with adaptive optics can evaluate photoreceptor function non-invasively in the living primate retina.

Deficiência de ferro: conseqüências biológicas e propostas de prevenção / Iron deficiency: biological consequences and prevention strategies

A deficiência de ferro é, isoladamente, a deficiência nutricional mais comum no mundo. Embora afete adultos, particularmente mulheres em idade reprodutiva e durante a gestação, as maiores vítimas são as crianças menores de dois anos de idade. Atinge principalmente a população de países em desenvolvimento que possui menor acesso a dietas equilibradas e a serviços de saúde e é mais exposta a condições sanitárias precárias. A deficiência de ferro tem impacto sobre a resposta imune, a capacidade de trabalho de adultos e o desenvolvimento cognitivo de crianças. O combate e a prevenção da deficiência de ferro é uma das prioridades na promoção da saúde. As estratégias para se alcançar este objetivo incluem a avaliação e correção da deficiência de ferro em gestantes, a estimulação ao aleitamento materno, a realização de profilaxia com sulfato ferroso oral em prematuros e recém nascidos de baixo peso, enriquecimento de alimentos e o envolvimento da comunidade.

Efeito de um novo produto fermentado de soja sobre os lípides séricos de homens adultos normocolesterolêmicos / Effect of a new fermented soy milk product on serum lipid levels in normocholesterolemic adult men

This study was undertaken to verify the effect of a daily intake of a new fermented soy milk produced with Enterococcus faecium and Lactobacillus jugurti on the serum lipid levels in normocholesterolemic middle-aged men. The study was randomized, double-blind and placebo-controlled and was performed for a period of 6 weeks. Forty-four normocholesterolemic healthy, male volunteers, aged 40-55 years old were randomly separated in two groups: The F-group received 200 ml of the fermented product daily and the P-group received 200 ml of placebo (chemically fermented). The blood samples were drawn initially and after 3 and 6 weeks and serum values for total cholesterol, HDL-cholesterol and triglyceride were determined. The LDL-cholesterol value was estimated. No significant changes in the fermented group (F) were observed for total cholesterol, LDL-cholesterol or triglyceride levels, while the HDL-cholesterol level was significantly higher (p < or = 0.05) after 6 weeks. The total cholesterol and LDL-cholesterol levels were significantly higher (p < or = 0.05) in the placebo group (P), but no changes were found for the HDL-cholesterol and triglyceride levels during the experimental period. In conclusion, the intake of 200 ml/day of the fermented soy milk, produced with E. faecium and L. jugurti, for 6 weeks, did not affect the serum total cholesterol and LDL-cholesterol, and led an increase of 10 per cent in the HDL-cholesterol level.

[Effect of iron salts addition on the sensory characteristics of soy yogurt]. / Efeito de diferentes sais de ferro sobre as características sensoriais do "iogurte" de soja.

The soy-yogurt was used as food vehicle due to its therapeutic and nutritional properties and lower cost. The aim of this work was to develop an enriched soy-yogurt with 12 mg of elementary iron/l, with suitable sensory and technological properties. Four iron sources were tested: FeSO4.7H2O, NaFeEDTA, Ferrochel and microencapsulated FeSO4.7H2O. The products were evaluated by fermentation time, pH, titratable acidity, viscosity, consistency, iron concentration and sensory properties (difference from the control and acceptance tests). Viscosity and consistency data were submitted to analysis of variance and Tukey's test. Difference from the control data was evaluated by analysis of variance and Dunnett's test and the acceptance test was evaluated by analysis of variance and Tukey's test. For all iron salts used in the enrichment process, only the FeSO4.7H2O did not work out because of the undesirable sensorial characteristics of the final products. The others sources used in the enrichment process (NaFeEDTA, Ferrochel and microencapsulated FeSO4.7H2O) did not alter the fermentation time, titratable acidity and sensory and reologics properties of the soy-yogurt.

Effects of a novel fermented soy product on the serum lipids of hypercholesterolemic rabbits.

OBJECTIVE: To assess the effect of a new feed soy product fermented by Enterococcus faecium and Lactobacillus jugurti on the serum lipid levels of rabbits with induced hypercholesterolemia. METHODS: Thirty-two rabbits were divided into 4 groups as follows: 1) control (C); 2) hypercholesterolemic (H); 3) hypercholesterolemic + fermented product (HPF); and 4) control + fermented product (CPF). The H and HPF groups were fed with a diet with 0.15% (p/p) cholesterol in the first 15 days. C and CPF groups received regular food preparation. The HPF and CPF groups received 10 mL daily of the fermented 30 days. Blood samples were drawn at the beginning of the study and at the 15th and 30th days. Concentrations of total cholesterol, HDL-cholesterol, and triglycerides were analyzed. RESULTS: After 15 days, the HPF group showed a total cholesterol concentration lower (18.4%) than that of the H group (p = 0.05), but this difference disappeared after 30 days. No change was observed in total cholesterol levels of C and CPF groups. After 15 days, the HDL-cholesterol was higher (17.8%) in the HPF group, but the triglyceride levels remained unchanged in all groups during the same period of time. CONCLUSION: The soy fermented product caused an 18.4% reduction in total cholesterol and a 17.8% increase in the HDL-fraction. It may, therefore, be a possible coadjutor in the treatment of hypercholesterolemia.

Generation of a novel A kinase anchor protein and a myristoylated alanine-rich C kinase substrate-like analog from a single gene.

A unique Drosophila gene encodes two novel signaling proteins. Drosophila A kinase anchor protein 200 (DAKAP200) (753 amino acids) binds regulatory subunits of protein kinase AII (PKAII) isoforms in vitro and in intact cells. The acidic DAKAP200 polypeptide (pI approximately 3.8) contains an optimal N-terminal myristoylation site and a positively charged domain that resembles the multifunctional phosphorylation site domain of vertebrate myristoylated alanine-rich C kinase substrate proteins. The 15-kilobase pair DAKAP200 gene contains six exons and encodes a second protein, DeltaDAKAP200. DeltaDAKAP200 is derived from DAKAP200 transcripts by excision of exon 5 (381 codons), which encodes the PKAII binding region and a Pro-rich sequence. DeltaDAKAP200 appears to be a myristoylated alanine-rich C kinase substrate analog. DAKAP200 and DeltaDAKAP200 are evident in vivo at all stages of Drosophila development. Thus, both proteins may play important physiological roles throughout the life span of the organism. Nevertheless, DAKAP200 gene expression is regulated. Maximal levels of DAKAP200 are detected in the pupal phase of development; DeltaDAKAP200 content is elevated 7-fold in adult head (brain) relative to other body parts. Enhancement or suppression of exon 5 excision during DAKAP200 pre-mRNA processing provides potential mechanisms for regulating anchoring of PKAII and targeting of cAMP signals to effector sites in cytoskeleton and/or organelles.

Characterization of the targeting, binding, and phosphorylation site domains of an A kinase anchor protein and a myristoylated alanine-rich C kinase substrate-like analog that are encoded by a single gene.

A novel Drosophila A kinase anchor protein, Drosophila A kinase anchor protein 200 (DAKAP200), is predicted to be involved in routing, mediating, and integrating signals carried by cAMP, Ca(2+), and diacylglycerol (Li, Z., Rossi, E. A., Hoheisel, J. D., Kalderon, D., and Rubin, C. S. (1999) J. Biol. Chem. 274, 27191-27200). Experiments designed to assess this hypothesis now (a) establish the function, boundaries and identity of critical amino acids of the protein kinase AII (PKAII) tethering site of DAKAP200; (b) demonstrate that residues 119-148 mediate binding with Ca(2+)-calmodulin and F-actin; (c) show that a polybasic region of DAKAP200 is a substrate for protein kinase C; (d) reveal that phosphorylation of the polybasic domain regulates affinity for F-actin and Ca(2+)-calmodulin; and (e) indicate that DAKAP200 is myristoylated and that this modification promotes targeting of DAKAP200 to plasma membrane. DeltaDAKAP200, a second product of the DAKAP200 gene, cannot tether PKAII. However, DeltaDAKAP200 is myristoylated and contains a phosphorylation site domain that binds Ca(2+)-calmodulin and F-actin. An atypical amino acid composition, a high level of negative charge, exceptional thermostability, unusual hydrodynamic properties, properties of the phosphorylation site domain, and a calculated M(r) of 38,000 suggest that DeltaDAKAP200 is a new member of the myristoylated alanine-rich C kinase substrate protein family. DAKAP200 is a potentially mobile, chimeric A kinase anchor protein-myristoylated alanine-rich C kinase substrate protein that may facilitate localized reception and targeted transmission of signals carried by cAMP, Ca(2+), and diacylglycerol.

An intramembrane modulator of the ErbB2 receptor tyrosine kinase that potentiates neuregulin signaling.

The ErbB2 receptor tyrosine kinase plays a critical role in a variety of developmental processes, and its aberrant activation may contribute to the progression of some breast and ovarian tumors. ASGP2, a transmembrane glycoprotein found on the surface of the highly metastatic ascites 13762 rat mammary adenocarcinoma cell line, is constitutively associated with ErbB2 in these cells and in mammary tissue from pregnant rats. Expression studies indicate that ASGP2 interacts directly and specifically with ErbB2 through one of its epidermal growth factor-like domains and that the co-expression of the two proteins in the same cell dramatically facilitates their direct stable interaction. Ectopic expression of ASGP2 in human melanoma tumor cells potentiates the response of endogenous ErbB2 to the neuregulin-1 growth factor. These observations point to a novel intramembrane mechanism for the modulation of receptor tyrosine kinase activity.

Sialomucin complex, a heterodimeric glycoprotein complex. Expression as a soluble, secretable form in lactating mammary gland and colon.

Ascites 13762 rat mammary adenocarcinoma cells express abundantly on their cell surfaces a heterodimeric glycoprotein complex composed of a sialomucin ascites sialoglycoprotein (ASGP)-1 and a transmembrane subunit ASGP-2. The latter, which contains two epidermal growth factor-like domains, binds the receptor tyrosine kinase p185(neu), suggesting that the complex is bifunctional as well as heterodimeric. Immunoblot analyses using monoclonal antibodies prepared against the complex demonstrate high levels of expression in rat lactating mammary gland and colon. Immunolocalization studies with anti-ASGP-2 indicate that ASGP-2 is present in these two tissues in the apical regions of secretory epithelial cells. Both mammary gland and colon contain a soluble, secretable form of ASGP-2, which is not found in the ascites cells; milk and mammary gland also have the membrane form. Immunoblot analyses using a COOH-terminal-specific polyclonal antibody indicate that the soluble form of ASGP-2 is missing its COOH-terminal domains. Both the soluble and membrane forms of ASGP-2 are similar to the membrane-associated form from the 13762 adenocarcinoma with respect to Mr, antigenicity, and association with ASGP-1. The presence of ASGP-1 in milk suggests that it is a candidate for the uncharacterized high Mr milk mucin, MUCX. ASGP-2 expression is up-regulated in mammary gland during pregnancy, because it is undetectable in virgin and early pregnant rats but abundant in the gland from late pregnant and lactating animals. However, compared with the lactating mammary gland, the 13762 ascites cells overexpress ASGP-2 by more than 100-fold, which may contribute to their malignancy. These combined results indicate that sialomucin complex is a unique secreted product in the mammary gland and colon, whose behavior is different from that in the mammary ascites tumors, and which may play important roles in mammary and intestinal physiology.

Monoclonal antibodies specific to sailfish serum albumin: development of an assay for the identification of fish species in the field.

Balb/c mice were immunized with albumin purified from sailfish (Istiophorus albicans) serum. Hybridomas were produced and screened by ELISA for reactivity with the purified albumins of sailfish, blue marlin (Makaira nigricans) and white marlin (Tetrapturus albidus). Monoclonal antibodies (MAbs) from 16 different clones exhibited activity against sailfish albumin. Thirteen of the MAbs showed cross-reactivity with the marlin species. Three MAbs exhibited distinct specificity for sailfish albumin. One of these species specific MAbs (M2D1) was conjugated to horseradish peroxidase (HRP) in order to construct an ELISA for identification of sailfish from serum. The ELISA for sailfish correctly identified eight sailfish from 26 billfish serum samples. The MAb-peroxidase conjugate was highly specific toward sailfish in that no reaction against heterologous species was detected.