Genome-wide analysis identifies gain and loss/change of function within the small multigenic insecticidal Albumin 1 family of Medicago truncatula.

BACKGROUND: Albumin 1b peptides (A1b) are small disulfide-knotted insecticidal peptides produced by Fabaceae (also called Leguminosae). To date, their diversity among this plant family has been essentially investigated through biochemical and PCR-based approaches. The availability of high-quality genomic resources for several fabaceae species, among which the model species Medicago truncatula (Mtr), allowed for a genomic analysis of this protein family aimed at i) deciphering the evolutionary history of A1b proteins and their links with A1b-nodulins that are short non-insecticidal disulfide-bonded peptides involved in root nodule signaling and ii) exploring the functional diversity of A1b for novel bioactive molecules. RESULTS: Investigating the Mtr genome revealed a remarkable expansion, mainly through tandem duplications, of albumin1 (A1) genes, retaining nearly all of the same canonical structure at both gene and protein levels. Phylogenetic analysis revealed that the ancestral molecule was most probably insecticidal giving rise to, among others, A1b-nodulins. Expression meta-analysis revealed that many A1b coding genes are silent and a wide tissue distribution of the A1 transcripts/peptides within plant organs. Evolutionary rate analyses highlighted branches and sites with positive selection signatures, including two sites shown to be critical for insecticidal activity. Seven peptides were chemically synthesized and folded in vitro, then assayed for their biological activity. Among these, AG41 (aka MtrA1013 isoform, encoded by the orphan TA24778 contig.), showed an unexpectedly high insecticidal activity. The study highlights the unique burst of diversity of A1 peptides within the Medicago genus compared to the other taxa for which full-genomes are available: no A1 member in Lotus, or in red clover to date, while only a few are present in chick pea, soybean or pigeon pea genomes. CONCLUSION: The expansion of the A1 family in the Medicago genus is reminiscent of the situation described for another disulfide-rich peptide family, the "Nodule-specific Cysteine-Rich" (NCR), discovered within the same species. The oldest insecticidal A1b toxin was described from the Sophorae, dating the birth of this seed-defense function to more than 58 million years, and making this model of plant/insect toxin/receptor (A1b/insect v-ATPase) one of the oldest known.

Severe systemic immune response syndrome, low plasma paraoxonase activity, and a new albumin species in a traumatized patient with Gaucher's disease.

INTRODUCTION: Gaucher's disease (GD) is an inborn error, autosomal recessive lysosomal lipid storage disorder characterized by the lack of the enzyme glucocerebrosidase. We observed some abnormalities in the plasma of a traumatized patient with GD. CASE REPORT: We report of a traumatized patient with GD that developed a severe systemic immune response during the course of an extended hospital stay. Plasma paraoxonase (PON) activity was assayed and found to be extremely low possibly due to the existence of GD in this particular patient. Also, a potentially novel post-translational modification (PTM) of albumin was noticed in the patient's plasma that coincided with enzyme replacement therapy (ERT) with Cerezyme. CONCLUSIONS: The decreased plasma PON activity measured might be a contributive factor in the development of an accentuated systemic immune response in a traumatized patient with GD. A modified albumin species could serve as a biomarker for ERT in Gaucher patients.

Pattern of ovarian protein synthesis and secretion during the reproductive cycle of Scotophilus heathi: synthesis of an albumin-like protein.

Ovarian proteins synthesized de novo and secreted in vitro were investigated during different stages of the reproductive cycle of Scotophilus heathi. We found increased ovarian protein synthesis and secretion during the recrudescence and the preovulatory periods, coinciding with two peaks of follicular development and steroidogenesis. The ovaries synthesized protein at a low rate during quiescence and the late phase of delayed ovulation. In vitro analysis using 35 S-methionine incorporation showed increased synthesis of 66 kDa protein during delayed ovulation, but secretion of this protein declined markedly during the preovulatory period. N-terminal sequencing of the 66 kDa protein showed that it shares 70% homology with human serum albumin. Immunocytochemistry indicated that this protein is found primarily in the granulosa cells of the follicles. Whether the increase in albumin production by the ovaries during delayed ovulation is associated with a hyperinsulinemic and hyperandrogenic condition requires further investigation.

Correlaçäo dos aspectos ultra-sonográficos com a classificaçäo clínica de child na cirrose hepática / Analyzing the echografpic alterations of the hepatic cirrhosis and matching them with the clinical classification of Child

Analisamos 165 pacientes com diagnóstico de cirrose hepática, avaliados clinicamente segundo a classificaçäo de Child, submetidos à ultra-sonografia abdominal e a dosagens séricas de alfa-fetoproteína (AFP), no período entre janeiro de 1994 e dezembro de 1995. O interesse deste estudo consiste em analisar as alteraçöes ecográficas da cirrose hepática e correlacioná-las com a classificaçäo clínica de Child. foi feita, também, análise da taxa de incidência do carcinoma hepatocelular e dos nódulos de regeneraçäo, correlacionando-os com os níveis séricos de AFP. Os resultados obtidos a partir deste estudo foram: do total dos pacientes examinados, 96 (58,2 por cento) pertenciam ao estádio Chid A, 61 (37.0 p/cento) ao Child B e 8 (4,8 por cento) ao Child C. Os níveis

Estudio comparativo de las técnicas de Machado Guerreiro y Elisa en el diagnóstico de infección chagásica / Compartive assay of Machado Guerreiro and Elisa techniques in diagnosis of chagas disease

Se procesaron 241 muestras de donantes voluntarios del banco de Sangre del Estado Zulia de sexo masculino y femenino, en edades comprendidas entre 18 y 30 años; a cada una de las cuales se le practicaron las pruebas de Elisa realizadas en el IVIC utilizando cepas puras del antígeno de T.cruzi, ELISA utilizando un kit comercial y Machado Guerreiro. Ambas técnicas realizadas en el Banco de Sangre del Estado Zulia. De los resultados obtenidos, con la técnica de Machado Guerreiro, 7 sueros resultaron positivos y 234 negativos, con ELISA (Banco de Sangre) 21 positivos y 220 negativos; con ELISA (IVIC) 32 positivos y 209 negativos. La comparación de Machado Guerreiro con ambas ELISA empleando el método de análisis Ji-Cuadrado modificado por Brand-Snedecor, reveló que las diferencias observadas entre los resultados de las técnicas fueron significativas. Evaluando los resultados ELISA (Banco de Sangre) y ELISA (IVIC) se demostró que no hubo diferencias significativas entre ambas pruebas. Los resultados de este estudio demostraron que ELISA es más sensible que Machado Guerreiro ya que detectó cantidades mínimas de anticuerpos específicos para antígenos chagásicos que no fueron detectados por Machado Guerreiro. El uso de la técnica ELISA amplía el rango de seguridad para las transfunsiones sanguíneas en relación a pacientes con infección chagásica

Experience with the use of human albumin in renal patients at the Kenyatta National Hospital.

In a 20-month period (May 1990 to December 1991), 34 patients with diuretic resistant oedema due to nephrotic syndrome, 4 patients with severe hypotension during haemodialysis and 3 patients with refractory ascites on chronic haemodialysis were treated with infusion of highly purified human placental albumin (HPHPA). In the 34 patients with resistant oedema due to nephrotic syndrome, infusion with HPHPA resulted in prompt diuresis, weight loss and normalisation of the blood urea nitrogen. These effects were maintained for three weeks after stopping the daily infusions, but started reverting to the pretreatment levels thereafter. Twelve of the 34 patients in whom prior attempts or renal biopsy had failed because of gross oedema, had this done following the resolution of oedema. In the 4 patients with severe hypotension (blood pressure, Systolic/Diastolic mmHg; Mean SD +/- 792 +/- 50 +/- 9) during haemodialysis the blood pressure rose to 135 +/- 11/87 +/- 8 following a rapid infusion of 100mls of 25% HPHPA and remained at that level during the course of haemodialysis. Three patients on chronic haemodialysis with refractory ascites were treated with four sessions of large-volume paracentesis. At each session they were infused with 200mls of 25% HPHPA over one hour. No clinically overt haemodynamic disturbance was noted thereafter and the patients were relieved of abdominal discomfort and respiratory difficulties for a period varying between 4 and 11 weeks before the ascites reaccumulated.

Human albumin genetic variants: an attempt at a classification of European allotypes.

The relative mobility of albumin and proalbumin genetic variants was estimated by means of cellulose acetate electrophoresis performed with three buffer systems at different pH (8.6, 5.0, and 6.9) after addition of a reference protein and dilution of sera. Numerous experiments using samples of reference variants corroborated the accuracy and reproducibility of this technique. The estimation of the variants' relative mobility at three pH allowed us to distinguish three fast-moving variants (Gent, Vanves, and Reading) and five slow-moving variants (Sondrio, Roma, Christchurch, Lille, and B) in the French population. The frequency of alloalbuminemia in this population is .0004 and is characterized by the high occurrence of albumin B and of the two proalbumin variants, Christchurch and Lille. In order to classify the variants of European origin, the methodology that we developed, owing to its more resolutive possibilities, should be employed as a first step in their identification until establishment of a structural nomenclature making mention of the amino acid substitution characterizing each variant.