Variability studies of allochthonous stone pine (Pinus pinea L.) plantations in Chile through nut protein profiling.

Stone pine (Pinus pinea) is characterized by low differentiation of growth parameters, high phenotypic plasticity and low genetic variability; detecting its diversity in introduced Chilean populations is therefore relevant for conservation and breeding programs. Here, variability among allochthonous Stone pine populations in Chile was explored using electrophoresis-based proteomic analysis of pine nuts. Cones from 30 populations distributed along a climatic gradient in Chile were surveyed and sampled, and proteins were extracted from seed flour using the TCA-acetone precipitation protocol. Extracts were subjected to SDS-PAGE and 2-DE for protein resolution, gel images captured, and spot or bands intensity quantified and subjected to statistical analysis (ANOVA, unsupervised Hierarchical Analysis Clustering and PLS regression). Protein yield ranged among populations from 161.7 (North populations) to 298.7 (South populations) mg/g dry weight. A total of 50 bands were resolved by SDS-PAGE in the 6.5-200 kDa Mr. range, of which 17 showed quantitative or qualitative differences, with 12 proteins identified. Pine nut extracts from the most distant populations were analyzed by 2-DE and a total of 129 differential spots were observed, out of which 13 were proposed as putative protein markers of variability. Out of the 129 spots, 118 proteins were identified after MALDI-TOF/TOF analysis. Identified proteins were classified into two principal categories: reserve and stress related. We provide the first protein map of P. pinea nuts. The use of a proteomic approach was useful to detect variability of Stone pine across three Chilean macrozones, with correlations between protein profiles and geoclimatic parameters, suggesting a new approach to study the variability of this species. BIOLOGICAL SIGNIFICANCE: This study presents the first protein map of Stone pine nuts, relevant for the advancement of protein characterization in pine nuts. Putative protein markers are proposed, evidencing that a proteomic approach may be useful to detect variability of Stone pine across Chilean macrozones, suggesting a new approach to study the variability of this species, which may also be extrapolated to other forest fruit species.

Generation of polyclonal antibodies against recombinant human glucocerebrosidase produced in escherichia coli

Deficiency of the lysosomal glucocerebrosidase (GCR) enzyme results in Gaucher's disease, the most common inherited storage disorder. Treatment consists of enzyme replacement therapy by the administration of recombinant GCR produced in Chinese hamster ovary cells. The production of anti-GCR antibodies has already been described with placenta-derived human GCR that requires successive chromatographic procedures. Here, we report a practical and efficient method to obtain anti-GCR polyclonal antibodies against recombinant GCR produced in Escherichia coli and further purified by a single step through nickel affinity chromatography. The purified GCR was used to immunize BALB/c mice and the induction of anti-GCR antibodies was evaluated by enzyme-linked immunosorbent assay. The specificity of the antiserum was also evaluated by western blot analysis against recombinant GCR produced by COS-7 cells or against endogenous GCR of human cell lines. GCR was strongly recognized by the produced antibodies, either as cell-associated or as secreted forms. The detected molecular masses of 59-66 kDa are in accordance to the expected size for glycosylated GCR. The GCR produced in E. coli would facilitate the production of polyclonal (shown here) and monoclonal antibodies and their use in the characterization of new biosimilar recombinant GCRs coming in the near future.

[Changes in the surgical approach of impacted lithiasis in the terminal choledochus]. / Cambios en la táctica quirúrgica de la litiasis impactada en el coledoco terminal.

Transduodenal sphincterotomy used to be was a common procedure when the surgeon couldn't remove stones from the terminal choledochus. We treated 6 patients in whom instead of a sphincterotomy, the stone was left in place and the duct drained with a T-tube. The first three cases were so managed because an emergency left no other option in the other three the decision was elective. Operative cholangiographies showed the impacted stones and in the postoperative cholangiographies, the stones had passed down to the stones had moved upward because of the decompression with the T-tube. The problem in all five patients with residual stone was easily solved. In one patient the stone was moshed down to the duodenum with biliary irrigation. The other four underwent transfistular extraction. Reviewing the morbility of transduodenal sphincterotomy we concluded that in an impacted stone it has currently few indications. In this era of "Biliary Perestroika" and specifically in emergencies or surgeons not fully experienced in transduodenal sphincterotomies, to put a T-tube and in the postoperative period complete others non surgical methods is an acceptable criteria. Also this approach can be applied to laparoscopic surgery when the operative cholangiography shows an unexpected biliary duct stone. The technique of leaving a transcistic catheter followed in the postoperative period by other therapeutic methods maintained in the discussion, could avoid a conversion to laparatomy or a laparoscopic choledochotomy, technique still in a developing period.

Cambios en la táctica quirurgica de la litiasis impactada en el colédoco terminal / Changes in the surgical technic of impacted lithiasis in the terminal choledochus

La papilotomía transduodenal era la técnica clásica cuando, por una coledocotomía no se podía extraer un cálculo del colédoco terminal. En este trabajo se analizó la evolución de 6 enfermos en los que en lugar de una papilotomía se dejó el cálculo y se colocó un tubo de Kehr. En los 3 primeros casos fue por necesidad y en los otros 3, una decisión electiva. Las colangiografías operatorias mostraron el cálculo impactado mientras que en las postoperatorias se había movilizado, tal vel por la descompensación y en un caso se había eliminado espontáneamente. De los 5 enfermos con la litiasis residual, en uno pasó al duodeno con una coledococlisis y en los otros 4 se extrajo por vía transfistular, sin morbilidad ni mortalidad. Después de analizar las complicaciones de la papilotomía transduodenal, creemos que en la actualidad esta operación tiene indicaciones muy restringidas frente a una litiasis coledociana impactada. En la época de la "Perestroika Biliar" y específicamente en casos de urgencia o con cirujanos poco entrenados, dejar un tubo de Kehr y luego completar el tratamiento con medios no quirúrgicos es una conducta muy aceptable. También esta táctica se puede extrapolar al hallazgo colangiográfico de un cálculo coledociano durante una colecistectomía laparoscópica. La conducta de un drenaje transcístico permite aplicar los tratamientos mencionados en la discusión y así evitar una conversión o tener que hacer una coledocotomía, procedimiento laparoscópico aún en desarrollo

Cambios en la táctica quirúrgica de la litiasis impactada en el coledoco terminal. / [Changes in the surgical approach of impacted lithiasis in the terminal choledochus]

Transduodenal sphincterotomy used to be was a common procedure when the surgeon couldnt remove stones from the terminal choledochus. We treated 6 patients in whom instead of a sphincterotomy, the stone was left in place and the duct drained with a T-tube. The first three cases were so managed because an emergency left no other option in the other three the decision was elective. Operative cholangiographies showed the impacted stones and in the postoperative cholangiographies, the stones had passed down to the stones had moved upward because of the decompression with the T-tube. The problem in all five patients with residual stone was easily solved. In one patient the stone was moshed down to the duodenum with biliary irrigation. The other four underwent transfistular extraction. Reviewing the morbility of transduodenal sphincterotomy we concluded that in an impacted stone it has currently few indications. In this era of [quot ]Biliary Perestroika[quot ] and specifically in emergencies or surgeons not fully experienced in transduodenal sphincterotomies, to put a T-tube and in the postoperative period complete others non surgical methods is an acceptable criteria. Also this approach can be applied to laparoscopic surgery when the operative cholangiography shows an unexpected biliary duct stone. The technique of leaving a transcistic catheter followed in the postoperative period by other therapeutic methods maintained in the discussion, could avoid a conversion to laparatomy or a laparoscopic choledochotomy, technique still in a developing period.

Cambios en la táctica quirurgica de la litiasis impactada en el colédoco terminal / Changes in the surgical technic of impacted lithiasis in the terminal choledochus

La papilotomía transduodenal era la técnica clásica cuando, por una coledocotomía no se podía extraer un cálculo del colédoco terminal. En este trabajo se analizó la evolución de 6 enfermos en los que en lugar de una papilotomía se dejó el cálculo y se colocó un tubo de Kehr. En los 3 primeros casos fue por necesidad y en los otros 3, una decisión electiva. Las colangiografías operatorias mostraron el cálculo impactado mientras que en las postoperatorias se había movilizado, tal vel por la descompensación y en un caso se había eliminado espontáneamente. De los 5 enfermos con la litiasis residual, en uno pasó al duodeno con una coledococlisis y en los otros 4 se extrajo por vía transfistular, sin morbilidad ni mortalidad. Después de analizar las complicaciones de la papilotomía transduodenal, creemos que en la actualidad esta operación tiene indicaciones muy restringidas frente a una litiasis coledociana impactada. En la época de la "Perestroika Biliar" y específicamente en casos de urgencia o con cirujanos poco entrenados, dejar un tubo de Kehr y luego completar el tratamiento con medios no quirúrgicos es una conducta muy aceptable. También esta táctica se puede extrapolar al hallazgo colangiográfico de un cálculo coledociano durante una colecistectomía laparoscópica. La conducta de un drenaje transcístico permite aplicar los tratamientos mencionados en la discusión y así evitar una conversión o tener que hacer una coledocotomía, procedimiento laparoscópico aún en desarrollo (AU)

Brida amniotica envolvendo o cordao umbilical em gestacao de termo. Revisao da literatura e apresentacao de um caso. / Amniotic bands round umbilical cord in full-term pregnancy. A review of literature and case report

O autor reviu a literatura de brida amniotica envolvendo o cordao umbilical na gestacao de termo, apresentando novo caso.Analisando os 42 casos relatados, com cinco sobrevidas fetais, conclui ser entidade rara, porem de elevado indice de obituario fetal

Maturidade fetal: estudo das celulas orangiofilas no liquido amniotico. / Fetal maturity: study of orangestaining cells in the amniotic fluid

Os autores estudaram 95 amostras de liquido amniotico obtidas de gestantes de termo, por amniocentese transvaginal ou transabdominal, e determinaram a porcentagem de celulas orangiofilas no liquido amniotico com o azul-de-nilo. Estabeleceram que em seu Servico, 20% (ou mais) indicaram maturidade fetal. A idade da gestacao e o peso fetal foram corretamente indicados, respectivamente, em 88% e 85% dos casos estudados