High genetic diversity in the harvestman Geraeocormobius sylvarum (Arachnida,Opiliones, Gonyleptidae) from subtropical forests in north‐eastern Argentina revealed by mitochondrial DNA sequences

This research was aimed to analyse the genetic diversity of Geraeocormobius sylvarum, a forest‐dwelling Neotropical harvestman with a disjunct distribution, separated by approximately 630 km of semi‐arid environments. The usefulness of a fragment of the cytochrome c oxidase subunit I (COI) mitochondrial gene as molecular marker was tested in 109 individuals. Results showed high levels of both haplotype and nucleotide diversity in populations corresponding to north‐eastern Argentina, the core area of the species range. A strong genetic structuring was detected, supported by both the phylogenetic trees and the haplotype network, with six identifiable haplogroups. Populations of the Yungas ecoregion did not show significant diversity levels, suggesting a putative recent introduction of the species into that region. The overall results suggest that the present genetic diversity of the species is consistent with past fragmentation events of the species range (in refuges?), probably during the Last Glacial Maximum. The COI gene was concluded to be a well‐suited marker to associate past environmental events with the high genetic diversity observed in this species.     

Structural characterization of ferric hemoglobins from three antarctic fish species of the suborder notothenioidei

Spontaneous autoxidation of tetrameric Hbs leads to the formation of Fe (III) forms, whose physiological role is not fully understood. Here we report structural characterization by EPR of the oxidized states of tetrameric Hbs isolated from the Antarctic fish species Trematomus bernacchii, Trematomus newnesi, and Gymnodraco acuticeps, as well as the x-ray crystal structure of oxidized Trematomus bernacchii Hb, redetermined at high resolution. The oxidation of these Hbs leads to formation of states that were not usually detected in previous analyses of tetrameric Hbs. In addition to the commonly found aquo-met and hydroxy-met species, EPR analyses show that two distinct hemichromes coexist at physiological pH, referred to as hemichromes I and II, respectively. Together with the high-resolution crystal structure (1.5 A) of T. bernacchii and a survey of data available for other heme proteins, hemichrome I was assigned by x-ray crystallography and by EPR as a bis-His complex with a distorted geometry, whereas hemichrome II is a less constrained (cytochrome b5-like) bis-His complex. In four of the five Antartic fish Hbs examined, hemichrome I is the major form. EPR shows that for HbCTn, the amount of hemichrome I is substantially reduced. In addition, the concomitant presence of a penta-coordinated high-spin Fe (III) species, to our knowledge never reported before for a wild-type tetrameric Hb, was detected. A molecular modeling investigation demonstrates that the presence of the bulkier Ile in position 67beta in HbCTn in place of Val as in the other four Hbs impairs the formation of hemichrome I, thus favoring the formation of the ferric penta-coordinated species. Altogether the data show that ferric states commonly associated with monomeric and dimeric Hbs are also found in tetrameric Hbs.     

Correlation of Cold Tolerance at Different Growth Stages in Rice

Cold injury which influence rice production in different rice growing countries occurs at various growth stages. The degree of cold injury depends on the air or water temperature, the cropping pattern, the growth stage of the rice and other factors. It is generally accepted that cold tolerance of rice at one stage is different from another stage. However, Okabe and Toriyama reported that varieties seem to respond similarly to cold temperature at different growth stages. Some varieties have been found to be tolerant at different growth stages. The purpose of this experiment is to find whether cold tolerance scores at different growth stages of different kinds of rice are correlated or not.     

Morphological and physiological differences between two morphotypes of Zostera noltii Hornem. from the south-western Iberian Peninsula

The morphological and physiological differences between two morphotypes of Z. noltii Hornem. were studied in the intertidal meadows on the south-western Iberian Peninsula (Palmones river estuary and Ria Formosa). A small-leaved morphotype (SM) grows mainly at high intertidal sites, meadow edges or in recently deposited sandbanks, whereas a large-leaved morphotype (LM) generally thrives in well-structured beds or in deeper places. This study deals with the morphological, biochemical and physiological differences between these morphotypes as well as the ecological implications of the occurrence of different morphotypes in the same meadow. Shoot length, leaf width, rhizome internode length, roots per node, root length, leaf nutrient and pigment contents, and photosynthetic rates of both morphotypes were compared. The below-ground architecture (root and rhizome complex) of both morphotypes was more developed in sites characterized by higher hydrodynamics and/or a lower nitrogen content in sediments. Both morphotypes showed similar values for photosynthetic efficiency, dark respiration rate and compensation irradiance. On the other hand, the net photosynthetic capacity was much greater (5-fold) for the SM. This difference could explain the greater growth rate and faster leaf turnover rate of the SM compared with the LM. The occurrence of the SM in newly settled areas (and in the meadow edges) could be explained on the basis of its higher growth rate, which would allow a faster spreading of the meadow and/or better recovery after burial resulting from stormy weathers. Received: 20 December 1998 / Received in revised form: 21 April 1999 / Accepted: 23 April 1999     

Both afferent and efferent nerves are implicated in cholecytokinin motor actions in the small intestine of the rat.

Cholecystokinin (CCK) regulates intestinal motility after being released by several luminal nutrients. However the mechanism of action of CCK is still not well known. The aim of our study was to establish the mechanism of action of CCK in the rat intestine using an in vivo model and focusing on the nervous pathways involved in the response as well as type of receptors. Anesthetized rats were prepared with two strain-gauges, in duodenum and jejunum, to record circular muscle motor activity. A group of animals was also prepared with a catheter to infuse capsaicin inside the duodenum. Responses to CCK-octapeptide (CCK-8) as well as to CCK agonists were studied. CCK-8 was also infused after CCK antagonists, atropine, hexamethonium or L-nitroarginine. Results show that duodenal response to CCK-8 is excitatory although inhibitory responses can be induced by gastrin. In the jejunum, CCK-8 induces an inhibitory response that is mediated by both CCK-A and -B receptors. Excitatory responses to CCK-8 are due to stimulation of preganglionic receptors while inhibitory responses are NO mediated through stimulation of postganglionic CCK-B receptors. Capsaicin locally applied in duodenal mucosa significantly decreased CCK-8 response, whereas mucosal exposure to lidocaine completely blocked CCK-8 response. In conclusion our results show that CCK response varies along the intestine according to the predominance of excitatory or inhibitory efferent innervation. Moreover, CCK-8 actions are mediated through both extrinsic and intrinsic afferent fibres.     

Cell cycle-dependent subcellular localization of exchange factor directly activated by cAMP

Epac belongs to a new family of proteins that can directly mediate the action of the intracellular second messenger cAMP by activating a downstream small GTPase Rap1. The Epac/Rap1 pathway represents a novel cAMP-signaling cascade that is independent of the cAMP-dependent protein kinase (PKA). In this study, we have used fluorescence microscopy to probe the intracellular targeting of Epac during different stages of the cell division cycle and the structural features that are important for Epac localization. Our results suggest Epac, endogenous or expressed as a green fluorescent protein fusion protein, is mainly localized to the nuclear membrane and mitochondria during interphase in COS-7 cells. Deletion mutagenesis analysis reveals that whereas the DEP domain is responsible for membrane association, the mitochondrial-targeting sequence is located at the N terminus. Although Epac predominantly exhibits perinuclear localization in interphase, the subcellular localization of Epac is cell cycle-dependent. Epac disassociates from the nuclear membrane and localizes to the mitotic spindle and centrosomes in metaphase. At the end of the cell cycle, Epac is observed to reassociate with the nuclear envelope and concentrate around the contractile ring. Furthermore, overexpression of Epac in COS-7 cells leads to an increase in multinuclear cell populations. These results suggest that Epac may play an important role in mitosis.     

Proteomic map of peripheral blood mononuclear cells

In the field of proteomics extensive efforts have been focused on the knowledge of proteins expressed by different cell types. In particular, enormous progress has been done in the characterization of blood cellular components. In this work, we have established a public 2-DE database for human peripheral blood mononuclear cells (PBMCs) proteins. Two hundred and forty-six spots corresponding to 174 different proteins have been identified on 2-DE gels from PBMCs isolated from six healthy individuals. All the identified proteins have been classified in thirteen categories on the basis of their differential functions or cellular localization and annotated at the http://physiology.unile.it/proteomics. The role of several proteins has been discussed in relation to their biological function. We intend to show the potentiality of PBMCs to investigate the proteomics changes possibly associated with a large number of pathologies such as autoimmune, neurodegenerative and cancer diseases.     

Conserved group-specific epitopes of the circumsporozoite proteins revealed by antibodies to synthetic peptides

The immunogenic properties of sporozoites are associated mainly with the circumsporozoite (CS) protein that covers the surface of mature sporozoites. This stage-specific protein has an immunodominant region with repetitive epitopes. Rabbits that are repeatedly immunized with sporozoites of Plasmodium knowlesi, a monkey malaria parasite, also recognize two synthetic peptides (N2 and C2) representing other polar domains of the CS protein. We show in this report that antibodies to the N2 and C2 synthetic peptides react not only with P. knowlesi but also with conserved regions of the surface membrane of other human, monkey, and rodent (but not avian) malaria sporozoites. Moreover, antibodies to N2 partially neutralize the infectivity of sporozoites of P. berghei, a rodent malaria parasite. In contrast, antibodies to synthetic peptides representing the repetitive epitope of P. knowlesi were strictly species specific.