The role of lateral and vertical herkogamy in the divergence of the blue- and red-flowered lineages of Lysimachia arvensis.

BACKGROUND AND AIMS: Herkogamy, or anther-stigma separation, is known to reduce self-pollen deposition, but little is known about the relative efficacy of different modes or conformations of herkogamy. We assessed the effectiveness of vertical versus lateral herkogamy in preventing or promoting self-pollen deposition in the annual herb Lysimachia arvensis, a plant with lineages that differ in flower colour, and in which flowers first display lateral and then vertical herkogamy. Because mating between the two lineages compromises fitness through the production of low-quality hybrid offspring, we tested the prediction that individuals sampled from sites occupied by both lineages should have flowers that promote autonomous self-pollen deposition and self-fertilization as a result of selection to reduce deleterious reproductive interference. METHODS: We characterized variation in herkogamy within and among 25 pure and mixed populations of L. arvensis in its European range and assessed the effectiveness of lateral versus vertical herkogamy in avoiding self-pollen deposition. RESULTS: Lateral herkogamy was more effective than vertical herkogamy in limiting self-pollen deposition. In the case of vertical herkogamy, only approach herkogamy was effective. Lineages showed consistent differences in herkogamy traits. In general, angles were smaller for blue than red flowers in most populations, and blue flowers showed approach herkogamy, while red flowers showed predominantly reverse herkogamy. In sympatry, the red lineage showed a reduction of both herkogamy traits while for the blue lineage only lateral herkogamy was reduced. CONCLUSIONS: Our results demonstrate that pollen deposition is affected not only by the degree but also the spatial conformation of herkogamy. They also highlight reduced herkogamy as a potential mechanism for promoting reproductive assurance under pollen limitation, as well as for avoiding reproductive interference between genetically divergent lineages.

Uncertain pollination environment promotes the evolution of a stable mixed reproductive system in the self-incompatible Hypochaeris salzmanniana (Asteraceae).

Background and aims: The transition from outcrossing to selfing is a repeated pattern in angiosperm diversification and according to general theory this transition should occur quickly and mixed reproductive systems should be infrequent. However, a large proportion of flowering plants have mixed reproductive systems, even showing inbreeding depression. Recently, several theoretical studies have shown that mixed mating systems can be stable, but empirical studies supporting these assumptions are still scarce. Methods: Hypochaeris salzmanniana, an annual species with populations differing in their self-incompatibility expression, was used as a study case to assess the stability of its mixed reproductive system. Here a descriptive study of the pollination environment was combined with measurements of the stability of the self-incompatibility system, outcrossing rate, reproductive assurance and inbreeding depression in four populations for two consecutive years. Key Results: The reproductive system of populations exhibited a geographical pattern: the proportion of plants decreased from west to east. Pollinator environment also varied geographically, being less favourable from west to east. The self-incompatibility expression of some populations changed markedly in only one year. After selfing, progeny was mainly self-compatible, while after outcrossing both self-incompatible and self-compatible plants were produced. In general, both reproductive assurance and high inbreeding depression were found in all populations and years. The lowest values of inbreeding depression were found in 2014 in the easternmost populations, which experienced a marked increase in self-compatibility in 2015. Conclusions: The mixed reproductive system of H. salzmanniana seems to be an evolutionarily stable strategy, with selfing conferring reproductive assurance when pollinator attendance is low, but strongly limited by inbreeding depression. The fact that the highest frequencies of self-compatible plants appeared in the environments most unfavourable to pollination suggests that these plants are selected in these sites, although high rates of inbreeding depression should impede the complete loss of self-incompatibility. In H. salzmanniana, year-to-year changes in the frequency of self-incompatible individuals are directly derived from the balance between reproductive assurance and inbreeding depression.

Pollinator response to yellow UV-patterned versus white UV-patternless flower dimorphism in Anemone palmata.

Flower colour polymorphisms are uncommon but widespread among angiosperms and can be maintained by a variety of balancing selection mechanisms. Anemone palmata is mostly yellow-flowered, but white-flowered plants coexist in some populations. We analysed the distribution of colour morphs of A. palmata across its range. We also characterised their colours and compared their vegetative and sexual reproductive traits, pollinator attention and fitness. The range of A. palmata is limited to the Western Mediterranean, while white-flowered plants are restricted to Portugal and SW Spain, where they occur at low proportions. Yellow flowers have a characteristic UV pattern, with a UV-absorbing centre and UV-reflecting periphery, which is absent in the white morph. Colour features of both morphs were highly delineated, making it easy for pollinators to distinguish them. Both morphs were protogynous, with the same duration of sexual stages, and the main floral traits related to pollinator attraction, apart from flower colour, were similar. Hymenoptera and Diptera were the main pollinators, showing preference for the yellow morph, clear partitioning of pollinator groups between the two colour morphs and a marked constancy to flower colour during foraging. Both morphs combined clonal propagation with sexual reproduction, but sexual reproductive potential was lower in white-flowered plants. Finally, female fitness was higher in the yellow morph. Pollinator partitioning and colour constancy could maintain this polymorphism, despite the lower visitation rate and fitness of white-flowered plants, which could facilitate their clonal propagation.

Isolation of a new Mexican strain of Bacillus subtilis with antifungal and antibacterial activities.

Although several strains of B. subtilis with antifungal activity have been isolated worldwide, to date there are no published reports regarding the isolation of a native B. subtilis strain from strawberry plants in Mexico. A native bacterium (Bacillus subtilis 21) demonstrated in vitro antagonistic activity against different plant pathogenic fungi. Under greenhouse conditions, it was shown that plants infected with Rhizoctonia solani and Fusarium verticillioides and treated with B. subtilis 21 produced augment in the number of leaves per plant and an increment in the length of healthy leaves in comparison with untreated plants. In addition, B. subtilis 21 showed activity against pathogenic bacteria. Secreted proteins by B. subtilis 21 were studied, detecting the presence of proteases and bacteriocin-like inhibitor substances that could be implicated in its antagonistic activity. Chitinases and zwittermicin production could not be detected. Then, B. subtilis 21 could potentially be used to control phytopathogenic fungi that infect strawberry plants.

Molecular phylogeny and systematics of the highly polymorphic Rumex bucephalophorus complex (Polygonaceae).

Rumex bucephalophorus is a very polymorphic species that has been subjected to various taxonomic studies in which diverse infraspecific taxa have been recognised on the basis of diaspore traits. In this study we used molecular markers (ITS and AFLP) to explore this remarkable diversity, to test previous hypotheses of classification, and attempt to explain biogeographic patterns. Results show that R. bucephalophorus forms a monophyletic group in which diversification began around 4.2 Mya, at the end of Messinian Salinity Crisis. The two molecular markers clearly show a deep divergence separating subsp. bucephalophorus from all other subspecific taxa, among which subsp. canariensis also constitutes a separate and well distinguishable unit. In contrast, subspecies hispanicus and subsp. gallicus constitute a monophyletic group in which three subgroups can be recognised: subsp. hispanicus, subsp. gallicus var. gallicus and subsp. gallicus var. subaegeus. However, these three subgroups are not clearly distinguished genetically or morphologically, so that in formal classification it would be preferable to treat them at the varietal level.

Pollinators, flowering phenology and floral longevity in two Mediterranean Aristolochia species, with a review of flower visitor records for the genus.

The pollination of Aristolochia involves the temporary confinement of visitors inside the flower. A literature review has shown that some species are visited by one or a few dipteran families, while others are visited by a wider variety of dipterans, but only some of these are effective pollinators. We observed flowering phenology and temporal patterns of pollinator attendance in diverse populations of Aristolochia baetica and A. paucinervis, two species that grow in SW Spain, frequently in mixed populations. The two species had overlapping floral phenologies, extended flowering periods and long-lived flowers. A. baetica attracted a higher number of visitors than A. paucinervis. Drosophilids and, to a lesser extent, phorids, were the main pollinators of A. baetica, whereas in A. paucinervis, phorids were the only pollinators. Attendance to A. paucinervis flowers by phorids in mixed populations was markedly lower than in pure populations. This effect was more evident in years with lower pollinator density. Our results suggest that A. baetica and A. paucinervis may compete for pollinators in mixed populations.

The interaction between Cistaceae and a highly specific seed-harvester ant in a Mediterranean scrubland.

We studied the interaction between the ant Goniomma kugleri and Cistaceae in a Cistus ladanifer-dominated scrubland, in southwestern Spain. We monitored seed harvesting, and studied ant preferences among Cistaceae seeds and their capture efficiencies for preferred seeds. For the stand of C. ladanifer, we estimated seed losses due to the ants. Harvesting was restricted to two seasons: mid-autumn to late winter, and late spring. Ant diet relied on Cistaceae seeds: during autumn and winter 90% of seeds returned to nests were of C. ladanifer, and the remaining fraction also comprised Cistaceae seeds. At this time, the ants harvested seeds directly from the plants. In late spring, the ant diet consisted of Tuberaria guttata s.l. seeds. Goniomma kugleri selectively collected Cistaceae seeds. For preferred species, seed removal rates at the colony level and seed capture times invested by individual workers were correlated with seed size. Because of shorter capture time and higher success frequency, capture efficiency in terms of number of seeds captured per unit time was higher for small-seeded species. Although each ant colony collected large numbers (up to 10(5)) of C. ladanifer seeds over the autumn-winter season, the impact of ant removal on the annual seed output was moderate, at around 20%. It is likely that, in C. ladanifer, the staggered seed release period, and the pulsed exposure of seed clumps in capsules through progressive locule dehiscence, effectively minimise seed losses to the ants.

Perianth organization and intra-specific floral variability.

Floral symmetry and fusion of perianth parts are factors that contribute to fine-tune the match between flowers and their animal pollination vectors. In the present study, we investigated whether the possession of a sympetalous (fused) corolla and bilateral symmetry of flowers translate into decreased intra-specific variability as a result of natural stabilizing selection exerted by pollinators. Average size of the corolla and intra-specific variability were determined in two sets of southern Spanish entomophilous plant species. In the first set, taxa were paired by family to control for the effect of phylogeny (phylogenetically independent contrasts), whereas in the second set species were selected at random. Flower size data from a previous study (with different species) were also used to test the hypothesis that petal fusion contributes to decrease intra-specific variability. In the phylogenetically independent contrasts, floral symmetry was a significant correlate of intra-specific variation, with bilaterally symmetrical flowers showing more constancy than radially symmetrical flowers (i.e. unsophisticated from a functional perspective). As regards petal fusion, species with fused petals were on average more constant than choripetalous species, but the difference was not statistically significant. The reanalysis of data from a previous study yielded largely similar results, with a distinct effect of symmetry on variability, but no effect of petal fusion. The randomly-chosen species sample, on the other hand, failed to reveal any significant effect of either symmetry or petal fusion on intra-specific variation. The problem of low-statistical power in this kind of analysis, and the difficulty of testing an evolutionary hypothesis that involves phenotypic traits with a high degree of morphological correlation is discussed.

Flower colour polymorphism in the Mediterranean Basin: occurrence, maintenance and implications for speciation.

Flower colour polymorphism (FCP) is the occurrence of at least two discrete flower colour variants in the same population. Despite a vast body of research concerning the maintenance and evolutionary consequences of FCP, only recently has the spatial variation in morph frequencies among populations been explored. Here we summarise the biochemical and genetic basis of FCP, the factors that have been proposed to explain their maintenance, and the importance of FCP and its geographic variation in the speciation process. We also review the incidence of FCP in the environmentally heterogeneous Mediterranean Basin. Nearly 88% of Mediterranean FCP species showed anthocyanin-based polymorphisms. Concerning the evolutionary mechanisms that contribute to maintain FCP, selection by pollinators is suggested in some species, but in others, selection by non-pollinator agents, genetic drift or gene flow are also found; in some cases different processes interact in the maintenance of FCP. We emphasise the role of both autonomous selfing and clonal reproduction in FCP maintenance. Mediterranean polymorphic species show mainly monomorphic populations with only a few polymorphic ones, which generate clinal or mosaic patterns of variation in FCP. No cases of species with only polymorphic populations were found. We posit that different evolutionary processes maintaining polymorphism the Mediterranean Basin will result in a continuum of geographic patterns in morph compositions and relative frequencies of FCP species.

[Factors associated with delayed hospital treatment for patients with acute cerebrovascular disease]. / Factores asociados a la demora del tratamiento hospitalario de los pacientes con enfermedad cerebrovascular aguda.

INTRODUCTION: Cerebrovascular diseases are an important cause of morbidity and mortality in the world and the effectiveness of thrombolytic therapy depends on early intervention. AIMS: To determine the time taken for patients with acute cerebrovascular events to reach hospital and to observe the variables associated to a longer delay. PATIENTS AND METHODS: We conducted a retrospective, descriptive study of 151 patients treated for acute cerebrovascular events between September 2003 and September 2004 at the Hospital Nacional Cayetano Heredia (Lima, Peru). We searched the records for the time taken to reach hospital, demographic variables and cerebrovascular risk factors, and we observed whether they exerted a significant effect on the arrival time. RESULTS: It was found that 29.8% of patients with an acute cerebrovascular event go to hospital within the first three hours; the mean arrival time is 51.11 hours. In contrast, 51% of patients go to hospital after more than 12 hours. Taking three and six hours as the cut-off points, uni and multivariate analyses showed ischemic stroke to be linked to a longer delay; a diagnosis of atrial fibrillation is associated to a shorter delay in arrival time in the multivariate analysis if a cut-off time of three hours is taken into account. CONCLUSIONS: Despite the high percentage of patients who take a long time to visit hospital following a cerebrovascular event, approximately a third of the patients would benefit from early treatment with thrombolytic therapy.

Polyembryony and apomixis in Eriotheca pubescens (Malvaceae - Bombacoideae).

Apomixis and adventitious polyembryony have been reported for several species of Bombacoideae, including Eriotheca pubescens, a tree species of the Neotropical savanna (Cerrado) areas in Brazil. However, the origin of polyembryonic seeds and their importance for the reproduction of the species remained to be shown. Here, we analyzed the early embryology of this species to establish the apomictic origin of extranumerary embryos. We also observed the geographic distribution of polyembryony in E. pubescens, and tested if apomixis was related to the source of pollen (self or cross) and population density. Moreover, we tested if polyembryonic apomictic embryos would develop normally into seedlings. In the observed seed primordia, after a relatively long quiescent period, the zygote developed into a sexual embryo concurrently with adventitious apomictic embryos which developed from nucellus cells. Adventitious embryos develop faster than sexual ones and are morphologically similar, so that 44 days after anthesis it was virtually impossible to distinguish and trace the fate of the sexual embryo. Polyembryony is widely distributed in populations some 400 km distant, and only one strictly monoembryonic individual was observed during the study. The number of embryos per seed varied between fruits and individuals but was significantly higher in seeds from cross-pollinations than from selfs, although fruit and seed set after crosses were much lower than after selfs. Embryo development into seedlings depended on their weight at germination, but polyembryonic seeds germinated and produced up to seven seedlings per seed in greenhouse conditions. Adventitious embryony and apomictic seedlings would explain the mostly clonal populations suggested by molecular studies.

A metastatic breast carcinoma presenting as autoimmune hemolytic anemia.

The authors describe the case of a 75-year-old female who was hospitalized for anemia of unknown origin. Physical examination revealed a swelling in the right mammary region, where a mastectomy scar was present from surgery for a breast carcinoma. On admission, laboratory tests disclosed anemia (Hb, 8.5 g/dl), with a reticulocyte count of 65,000/mm3 and slightly increased bilirubin. Immunohematologic study revealed the presence of a red cell autoantibody with anti-D specificity in the serum and in the eluate from the patient's erythrocytes. A biopsy of the swelling was performed and histologic examination showed the presence of metastatic cells of breast carcinoma. The patient was given chemotherapy and radiotherapy. At this writing the anemia was absent, the immunohematologic study was negative, the swelling was greatly reduced, and no other metastatic lesions of breast carcinoma were present.

Use of plastic films in grain stores.

A laboratory study of wheat stored at two different levels of moisture content with and without presence of Sitophilus zeamais Motschulsky was performed. Flexible PVC and polythene films as seals were used in order to examine their effectiveness to maintain the wheat quality by comparing changes in moisture content, germination and weight loss. The PVC film showed a high water vapor permeability and it behaved as the permeable muslin control, keeping the grain quality in all tested conditions. The polythene film showed a low water vapor permeability and it allowed the insects development. It seems that plastic films may be useful in grain stores bearing in mind their properties, adequate sealing and protecting them from insect damage, besides they should be selected according to environmental conditions.

Detection of diabetic foot hyperthermia by infrared imaging.

In diabetic foot, the occurrence of an ulcer is often associated with hyperthermia. Hyperthermia is defined as a temperature greater than 2.2°C in a given region of one of the foot compared to the temperature of the same region of the contralateral foot. Unfortunately, hyperthermia is not yet assessed in current diabetic foot therapy. In this paper, we propose an easy way to detect a possible hyperthermia by using an infrared camera. A specific acquisition protocol of the thermal images is proposed. A dedicated image analysis is developed: it is composed of a contour detection of the 2 feet using the Chan and Vese active contour method associated to the ICP rigid registration technique. Among 85 type II diabetes persons recruited in the Dos de Mayo hospital in Lima, Peru, 9 individuals show significant hyperthermia. It is expected that the new possibility of detecting hyperthermia in hospitals or in diabetic health centers which is now available, thanks to the proposed method, will help in reducing foot ulcer occurrence for diabetic persons.

Persistently low fruiting success in the Mediterranean pipevine Aristolochia baetica (Aristolochiaceae): a multi-year study.

Low fruit set is common in many plant species and may be caused by a variety of factors, such as predation, resource limitation or deficient pollination, or it may be an evolutionary strategy. In this paper, we investigate factors that affect fruit set in Aristolochia baetica (Aristolochiaceae), a Mediterranean pipevine found in southwest Spain. Fruit production was monitored in two populations over 4 years (2002-2005), and the causes of flower or fruit loss were determined. Experimental hand-pollinations were performed, and germinated pollen grains on the stigmas of open-pollinated flowers were quantified. Fruit set was always very low (4-14%). Floral abscission initially reduced reproductive output by more than 50%; then herbivory (6-12%) and fruit abortion (8-26%) caused further reductions. Given that the number of efficiently pollinated flowers was always higher than that of ripe fruits, and that xenogamous hand-pollination did not increase fruit set in relation to open-pollination, the final fruit production of A. baetica seems not to be pollen-limited. Fruit abortion of effectively pollinated flowers supports the idea that resource availability limits fruit set. In A. baetica, fruit abortion could lead to mate selection of the best quality fruits. Moreover, the initiated fruits that finally abort could also satiate predators, contributing to increase progeny fitness.

Functional andromonoecy in Euphorbia (Euphorbiaceae).

The occurrence of cyathia containing staminate flowers but lacking a pistillate flower was studied in 17 species of Euphorbia. Male cyathia were found in the majority of species studied (88.2%) giving functional andromonoecy. In the male cyathia, the pistillate flower is generally totally absent, but sometimes a vestigial pistillate flower with a non-functional ovary is present. The proportion of male cyathia varied at both the population and species level. The position of male cyathia within the inflorescence showed a constant pattern among species: the proportion of male cyathia decreased from the first to the last levels of the pleiochasia. In general, perennial species had significantly higher proportions of male cyathia than annual species (mean 20 and 2.3%, respectively). In annual species there was a trend for production of male cyathia only in the first level of the inflorescence, whereas in perennials production up to the fourth level of the inflorescence was usual. Functional andromonoecy is common in Euphorbia and represents a new sex segregation in the genus. The selective forces causing this secondary sex segregation in Euphorbia, such as improved pollination or increased outcrossing, are discussed.

Apical pattern of fruit production in the racemes of Ceratonia siliqua (Leguminosae: Caesalpinioideae): role of pollinators.

Fruit production and arrangement within the raceme were studied in two dioecious populations of Ceratonia siliqua (Leguminosae: Caesalpinioideae), an arboreal species that produces caulogenous racemes (emerging only from the old branches) with numerous flowers. Fruit production per raceme was low and similar between years and populations and even between individuals. During flowering, there were considerable flower losses from predation and lack of pollination. A mean of nine flowers per raceme began the transformation into fruits, of which 77% aborted. The final fruit production per raceme increased significantly following hand pollination, but was always very much lower than the availability of flowers in the raceme. The results suggest that fruit production of each raceme is limited by both availability of resources and a deficient pollination. In racemes setting fruit arrangement follows a definite pattern that remains constant between years and populations: fruit production was significantly higher in the apical zone of the raceme and lower in the basal zone. The pollinators of C. siliqua (flies and wasps) showed a clear preference for beginning their visits at the apex of a raceme. As a result, the pollen load deposited on the stigmas decreased from apex to base of the raceme. In most of the flowers situated in the central and basal zone of the raceme, the number of pollen grains deposited on their stigmas was lower than the number of their ovules. The high number of seeds in developed fruits suggests that the plant selectively aborts flowers that receive a smaller pollen load. The results indicate that the final pattern of fruit arrangement within the raceme is a direct result of pollinator activity.

Pernicious anemia, vitiligo and positive antiglobulin test: an unusual association.

The authors present the case of a patient with concomitant pernicious anemia and positive antiglobulin test. After discussing previous cases in the literature and pointing out the probably simultaneous occurrence of the two, the authors comment on the persistence of a positive antiglobulin test after remission of the pernicious anemia and the association with vitiligo, which has been sometimes described as associated with autoimmune hemolytic anemia and more often with pernicious anemia, but not with both. The patient also had diabetes mellitus and a persistently false positive test for syphilis.