Chemical Tools of Octopus maya during Crab Predation Are Also Active on Conspecifics.

Octopus maya is a major socio-economic resource from the Yucatán Peninsula in Mexico. In this study we report for the first time the chemical composition of the saliva of O. maya and its effect on natural prey, i.e. the blue crab (Callinectes sapidus), the crown conch snail (Melongena corona bispinosa), as well as conspecifics. Salivary posterior glands were collected from octopus caught by local fishers and extracted with water; this extract paralyzed and predigested crabs when it was injected into the third pereiopod. The water extract was fractionated by membrane ultrafiltration with a molecular weight cut-off of 3 kDa leading to a metabolic phase (>3 kDa) and a neurotoxic fraction (<3 kDa). The neurotoxic fraction injected in the crabs caused paralysis and postural changes. Crabs recovered to their initial condition within two hours, which suggests that the effects of the neurotoxic fraction were reversible. The neurotoxic fraction was also active on O. maya conspecifics, partly paralyzing and sedating them; this suggests that octopus saliva might be used among conspecifics for defense and for reduction of competition. Bioguided separation of the neurotoxic fraction by chromatography led to a paralysis fraction and a relaxing fraction. The paralyzing activity of the saliva was exerted by amino acids, while the relaxing activity was due to the presence of serotonin. Prey-handling studies revealed that O. maya punctures the eye or arthrodial membrane when predating blue crabs and uses the radula to bore through crown conch shells; these differing strategies may help O. maya to reduce the time needed to handle its prey.

Complexes of D-type cyclins with CDKs during maize germination.

The importance of cell proliferation in plant growth and development has been well documented. The majority of studies on basic cell cycle mechanisms in plants have been at the level of gene expression and much less knowledge has accumulated in terms of protein interactions and activation. Two key proteins, cyclins and cyclin-dependent kinases (CDKs) are fundamental for cell cycle regulation and advancement. Our aim has been to understand the role of D-type cyclins and type A and B CDKs in the cell cycle taking place during a developmental process such as maize seed germination. Results indicate that three maize D-type cyclins-D2;2, D4;2, and D5;3-(G1-S cyclins by definition) bind and activate two different types of CDK-A and B1;1-in a differential way during germination. Whereas CDKA-D-type cyclin complexes are more active at early germination times than at later times, it was surprising to observe that CDKB1;1, a supposedly G2-M kinase, bound in a differential way to all D-type cyclins tested during germination. Binding to cyclin D2;2 was detectable at all germination times, forming a complex with kinase activity, whereas binding to D4;2 and D5;3 was more variable; in particular, D5;3 was only detected at late germination times. Results are discussed in terms of cell cycle advancement and its importance for seed germination.

Protein phosphorylation regulates in vitro spinach chloroplast petD mRNA 3'-untranslated region stability, processing, and degradation.

RNA-binding proteins (RNPs) participate in diverse processes of mRNA metabolism, and phosphorylation changes their binding properties. In spinach chloroplasts, 24RNP and 28RNP are associated with polynucleotide posphorylase forming a complex on charge of pre-mRNA 3'-end maturation. Here, we tested the hypothesis that the phosphorylation status of 24RNP and 28RNP, present in a spinach chloroplast mRNA 3'-UTR processing extract (CPE), controls the transition between petD precursor stabilization, 3'-UTR processing, and RNA degradation in vitro. The CPE processed or stabilized petD precursor depending on the ATP concentration present in an in vitro 3'-UTR processing (IVP) assay. These effects were also observed when ATP was pre-incubated and removed before the IVP assay. Moreover, a dephosphorylated (DP)-CPE degraded petD precursor and recovered 3'-UTR processing or stabilization activities in an ATP concentration dependent manner. To determine the role 24/28RNP plays in regulating these processes a 24/28RNP-depleted (Δ24/28)CPE was generated. The Δ24/28CPE degraded the petD precursor, but when it was reconstituted with recombinant non-phosphorylated (NP)-24RNP or NP-28RNP, the precursor was stabilized, whereas when Δ24/28CPE was reconstituted with phosphorylated (P)-24RNP or P-28RNP, it recovered 3'-UTR processing, indicating that 24RNP or 28RNP is needed to stabilize the precursor, have a redundant role, and their phosphorylation status regulates the transition between precursor stabilization and 3'-UTR processing. A DP-Δ24/28CPE reconstituted or not with NP-24/28RNP degraded petD precursor. Pre-incubation of DP-Δ24/28CPE with NP-24/28RNP plus 0.03 mM ATP recovered 3'-UTR processing activity, and its reconstitution with P-24/28RNP stabilized the precursor. However, pre-incubation of DP-Δ24/28CPE with 0.03 mM ATP, and further reconstitution with NP-24/28RNP or P-24/28RNP produced precursor stability instead of RNA degradation, and RNA processing instead of precursor stability, respectively. Moreover, in vitro phosphorylation of CPE showed that 24RNP, 28RNP, and other proteins may be phosphorylated. Altogether, these results reveal that phosphorylation of 24RNP, 28RNP, and other unidentified CPE proteins mediates the in vitro interplay between petD precursor stability, 3'-UTR processing, and degradation, and support the idea that protein phosphorylation plays an important role in regulating mRNA metabolism in chloroplast.

The family of maize D-type cyclins: genomic organization, phylogeny and expression patterns.

Cyclin proteins, associated to cyclin-dependent kinases (CDKs), play fundamental roles in cell cycle control as they constitute a very important driving force to allow cell cycle progression. D-type cyclins (CycDs) are important both for interpreting external mitogenic signals and in the control of the G1 phase. The maize (Zea mays) genome appears to contain at least 17 different CycD genes, and they fall into the subgroups previously described for other plants. Maize CycDs have been named according to identity percentages of the corresponding orthologs in rice and Arabidopsis. In silico analysis confirmed the presence of characteristic cyclin domains in each maize CycD gene and showed that their genomic organization is similar to their orthologs in rice and Arabidopsis. The expression of maize CycD genes was followed in seeds, during germination in the presence/absence of exogenously added hormones, and also in different plantlet tissues (mesocotyl, root tips and first leaf). Most cyclins were expressed in germinating seeds and at least in one of the plantlet tissues tested; almost all of the detected cyclins show an accumulating pattern of mRNA along germination (0-24 h) and higher levels in root tissue. Interestingly, some cyclins show high levels in non-proliferating tissues as leaf. Addition of auxins or cytokinins does not seem to importantly modify transcript levels; on the other hand, addition of abscisic acid repressed the expression of several cyclins. The role of each CycD during germination and plant growth and its interaction with other cell cycle proteins becomes a topic of the highest interest.

Colostomía perineal en el carcinoma recto vaginal / Perineal colostomy in recto-vaginal carcinoma

Se realizó una variante quirúrgica en una paciente con cáncer recto-vaginal, con infiltración del tercio inferior y cara lateral derecha del recto, muy próximo al esfínter del ano. La técnica consistió, previo diagnóstico por biopsia, en realizar excérecis del tumor que comprometió al parapeto derecho y el tercio inferior de vagina, terminando con una colostomía perineal, conservando algunas fibras del esfínter del ano. Se dejó drenaje en ambos lados y se reconstruyó resto de la vagina. Se logró buena continencia rectal del paciente, con buen estado de satisfacción. No hubo complicaciones y actualmente defeca sin dificultad. Ha tenido seguimiento por consulta de Oncoginecología del Hospital III Congreso de Pinar del Río. La intervención se llevó a cabo en una hora.

A surgical alternative was performed in a patient suffering from rectovaginal cancer infiltrating the lower third and lateral right aspect of the rectum, so closed to anal sphincter. Operative technique consisted of a previous diagnosis by biopsy, besides, exceresis of the tumor that compromised right pararectum and lower third of vagina, ending in perineal colostomy preserving some fibers of anal sphincter. Drainage was performed in both sides and the rest of the vagina was reconstructed. A good rectal continence and well being were obtained, observing no complications, the patient defecates without difficulties, this case is followed up at "III Congreso Hospital" in Pinar del Rio. Operative procedures lasted an hour.

Maize replicative alpha-type DNA polymerase: separation of polymerase and primase activities and recognition of primase subunits.

DNA polymerase and DNA primase activities in the maize alpha-type DNA polymerase 2 were dissociated and DNA polymerase-free DNA primase was studied. DNA primase synthesized primers that were 8-34 nucleotides long, with more intense bands at 15-17 nucleotides in length. DNA polymerase 1 (a putative delta-type enzyme) or DNA polymerase 2 were assayed after template-priming with purified DNA primase and showed a differential use of templates: whereas DNA polymerase 2 used a polydT template more efficiently than a natural template, DNA polymerase 1 used both of them poorly. The molecular size of DNA primase was estimated to be 68 kDa by gel filtration, western blotting and by a DNA primase 'trapping' assay.