Hybridization and adaptation to introduced balloon vines in an Australian soapberry bug.

Contemporary adaptation of plant feeding insects to introduced hosts provides clear cases of ecologically based population divergence. In most cases the mechanisms permitting rapid differentiation are not well known. Here we study morphological and genetic variation associated with recent shifts by the Australian soapberry bug Leptocoris tagalicus onto two naturalized Neotropical balloon vines, Cardiospermum halicacabum and C. grandiflorum that differ in time since introduction. Our results show that these vines have much larger fruits than the native hosts (Whitewood tree -Atalaya hemiglauca- and Woolly Rambutan -Alectryon tomentosus-) and that bugs living on them have evolved significantly longer beaks and new allometries. Genetic analyses of mitochondrial haplotypes and amplified fragment length polymorphic (AFLP) markers indicate that the lineage of bugs on the annual vine C. halicacabum, the older introduction, is intermediate between the two subspecies of L. tagalicus found on native hosts. Moreover, where the annual vine and Whitewood tree co-occur, the morphology and genomic composition of the bugs are similar to those occurring in allopatry. These results show that hybridization provided the genetic elements underlying the strongly differentiated 'Halicacabum bugs'. In contrast, the bugs feeding on the recently introduced perennial balloon vine (C. grandiflorum) showed no evidence of admixture, and are genetically indistinguishable from the nearby populations on a native host.

Post-mating prezygotic barriers to gene exchange between hybridizing field crickets.

Studies of sexual selection in speciation have traditionally focused on mate preference, with less attention given to traits that act between copulation and fertilization. However, recent work suggests that post-mating prezygotic barriers may play an important role in speciation. Here, we evaluate the role of such barriers in the field crickets, Gryllus firmus and Gryllus pennsylvanicus. Gryllus pennsylvanicus females mated with G. firmus males produce viable, fertile offspring, but when housed with both species produce offspring sired primarily by conspecifics. We evaluate patterns of sperm utilization in doubly mated G. pennsylvanicus females and find no evidence for conspecific sperm precedence. The reciprocal cross (G. firmus female × G. pennsylvanicus male) produces no progeny. Absence of progeny reflects a barrier to fertilization rather than reduced sperm transfer, storage or motility. We propose a classification scheme for mechanisms underlying post-mating prezygotic barriers similar to that used for premating barriers.

Population genetic structure of sexual and parthenogenetic damselflies inferred from mitochondrial and nuclear markers.

It has been postulated that obligate asexual lineages may persist in the long term if they escape from negative interactions with either sexual lineages or biological enemies; and thus, parthenogenetic populations will be more likely to occur in places that are difficult for sexuals to colonize, or those in which biological interactions are rare, such as islands or island-like habitats. Ischnura hastata is the only known example of natural parthenogenesis within the insect order Odonata, and it represents also a typical example of geographic parthenogenesis, as sexual populations are widely distributed in North America, whereas parthenogenetic populations of this species have only been found at the Azores archipelago. In order to gain insight in the origin and distribution of parthenogenetic I. hastata lineages, we have used microsatellites, mitochondrial and nuclear DNA sequence data, to examine the population genetic structure of this species over a wide geographic area. Our results suggest that sexual populations of I. hastata in North America conform to a large subdivided population that has gone through a recent spatial expansion. A recent single long distance dispersal event, followed by a demographic expansion, is the most parsimonious hypothesis explaining the origin of the parthenogenetic population of this species in the Azores islands.

Population assignment tests uncover rare long-distance marine larval dispersal events.

Long-distance dispersal (LDD) is consequential to metapopulation ecology and evolution. In systems where dispersal is undertaken by small propagules, such as larvae in the ocean, documenting LDD is especially challenging. Genetic parentage analysis has gained traction as a method for measuring larval dispersal, but such studies are generally spatially limited, leaving LDD understudied in marine species. We addressed this knowledge gap by uncovering LDD with population assignment tests in the coral reef fish Elacatinus lori, a species whose short-distance dispersal has been well-characterized by parentage analysis. When adults (n = 931) collected throughout the species' range were categorized into three source populations, assignment accuracy exceeded 99%, demonstrating low rates of gene flow between populations in the adult generation. After establishing high assignment confidence, we assigned settlers (n = 3,828) to source populations. Within the settler cohort, <0.1% of individuals were identified as long-distance dispersers from other populations. These results demonstrate an exceptionally low level of connectivity between E. lori populations, despite the potential for ocean currents to facilitate LDD. More broadly, these findings illustrate the value of combining genetic parentage analysis and population assignment tests to uncover short- and long-distance dispersal, respectively.

MC1R-dependent, melanin-based colour polymorphism is associated with cell-mediated response in the Eleonora's falcon.

Colour polymorphism in vertebrates is usually under genetic control and may be associated with variation in physiological traits. The melanocortin 1 receptor (Mc1r) has been involved repeatedly in melanin-based pigmentation but it was thought to have few other physiological effects. However, recent pharmacological studies suggest that MC1R could regulate the aspects of immunity. We investigated whether variation at Mc1r underpins plumage colouration in the Eleonora's falcon. We also examined whether nestlings of the different morphs differed in their inflammatory response induced by phytohemagglutinin (PHA). Variation in colouration was due to a deletion of four amino acids at the Mc1r gene. Cellular immune response was morph specific. In males, but not in females, dark nestling mounted a lower PHA response than pale ones. Although correlative, our results raise the neglected possibility that MC1R has pleiotropic effects, suggesting a potential role of immune capacity and pathogen pressure on the maintenance of colour polymorphism in this species.

[Structure of the arachnoid layer of the human spinal meninges: a barrier that regulates dural sac permeability]. / Morfología de la lámina aracnoidea espinal humana. Barrera que limita la permeabilidad del saco dural.

OBJECTIVES: Drugs injected into the epidural space are known to penetrate the subarachnoid space by simple diffusion through the dural sac. We aimed to study the cellular ultrastructure of the arachnoid membrane and the type of intercellular junctions responsible for creating the barrier that regulates the passage of drugs through the dural sac in humans. MATERIAL AND METHODS: Fourteen tissue samples of arachnoid membrane were taken from 2 patients during procedures that required opening the lumbar dural sac. The samples were treated with glutaraldehyde, osmium tetroxide, ferrocyanide and acetone, and then embedded in resin. Ultrathin sections were stained with lead citrate for examination by transmission electron microscopy. RESULTS: The arachnoid membrane was 35 to 40 microm thick. The outer surface contained neurothelial cells (dural border cells) along the subdural compartment, while the internal portion was made up of a plane 5 to 8 microm thick with 4 to 5 arachnoid cells overlapping to form a barrier layer. The intercellular spaces on this plane were 0.02 to 0.03 microm wide; the arachnoid cells were bridged by specialized junctions (desmosomes and other tight junctions). CONCLUSIONS: Structural features of the arachnoid cells provide a barrier within the human dural sac. They occupy only the internal portion of the arachnoid membrane. Specialized intercellular junctions explain the selective permeability of this membrane.

Clinical implications of epidural fat in the spinal canal. A scanning electron microscopic study.

BACKGROUND AND OBJECTIVES: This review of articles summarizes recent developments in relation to fat located in the epidural space and also in dural sleeves of spinal nerve roots in order to improve our understanding of the clinical effects of the epidural blockade. METHOD: Medline search was carried cross-matching of the following words: "epidural fat", "epidural space", "adipose tissue" and "fat cells" from 1966 to 2008 in which articles referring to different pathologies that alter the epidural fat were also reviewed. Techniques used by different authors included the use of samples from dissections, cryomicrotome sections, as well as light and electron microscopy. RESULTS: Fat in the epidural space has a metameric distribution along the spinal canal that can be altered in some pathological conditions. Epidural fat is not evenly distributed. At cervical level fat is absent while in the lumbar region, fat in the anterior and posterior aspects of the epidural space forms two unconnected structures. Fat cells are found also in the thickness of dural sleeves enveloping spinal nerve roots but not in the region of the dural sac. Epidural lipomatosis is characterized by an increase in epidural fat content. When a patient has a combination of kyphosis and scoliosis of the spine, the epidural fat distributes asymmetrically. Spinal stenosis is frequently accompanied by a reduction in the amount of epidural fat around the stenotic area. CONCLUSIONS: The epidural space contains abundant epidural fat that distributes along the spinal canal in a predictable pattern. Fat cells are also abundant in the dura that forms the sleeves around spinal nerve roots but they are not embedded within the laminas that form the dura mater of the dural sac. Drugs stored in fat, inside dural sleeves, could have a greater impact on nerve roots than drugs stored in epidural fat, given that the concentration of fat is proportionally higher inside nerve root sleeves than in the epidural space, and that the distance between nerves and fat is shorter. Similarly, changes in fat content and distribution caused by different pathologies may alter the absorption and distribution of drugs injected in the epidural space.

[Discrepancy between clinical and radiologic manifestations of an epidural hematoma after catheterization]. / Discordancia clínico-radiológica y resolución espontánea de un hematoma postcateterización epidural.

Neuraxial techniques are considered safe if certain guidelines are followed, but they are not risk free. We report the case of an 81-year-old woman with an invasive bladder tumor who underwent radical cystectomy with a Bricker-type procedure. General anesthesia was used and epidural analgesia was also provided for surgical and postoperative pain management. Late in the postoperative recovery period a large epidural hematoma was diagnosed based on radiologic signs of spinal cord compression, in the absence of symptoms other than mild and progressive back pain that developed after extubation. The surgeon decided against emergency surgery to reduce compression. Symptoms resolved gradually, and a magnetic resonance image 45 days after discharge confirmed that the hematoma was smaller. In addition to the usual safety recommendations for epidural anesthesia with regard to drugs that alter hemostasis, it is important to bear in mind circumstances that have pharmacokinetic repercussions and that increase risk. Lower back pain can be a warning sign. Some cases may resolve spontaneously.

[Ziconotide: an innovative alternative for intense chronic neuropathic pain]. / Ziconotide: una alternativa innovadora en el dolor crónico neuropático intenso.

INTRODUCTION: Intense chronic pain is a very important health problem, as it has a high prevalence (5-10%), a multifactorial aetiology and its management is very often a very complex affair. Treatment of severe cases sometimes requires interventional approaches, such as continuous intrathecal infusion of opioids. CASE REPORT: We report the case of a 38-year-old female with intense neuropathic pain in the lower back and the lower limbs secondary to three operations on the L5-S1 lumbar segment. After implementing several different pharmacological regimes involving both oral and implanted systems (spinal cord stimulation and subarachnoid infusion pump with different pharmacological combinations) with no clinical improvement, intrathecal infusion with ziconotide was included in the protocol. CONCLUSIONS: Ziconotide is the first specific neuronal blocker that acts on the calcium channel by blocking the N-type voltage-dependent calcium channels. It is a new non-opioid analgesic with approved indication in the treatment of intense chronic pain, in patients who require intrathecal analgesics and are refractory to other analgesic treatments. Therefore, we shall have to consider this drug as a therapeutic alternative in patients do not experience sufficient relief with the pharmacological agents and means currently available to treat them.

[Fat inside the dural sheath of lumbar nerve roots in humans]. / Grasa dentro de los manguitos durales de las raíces nerviosas de la columna lumbar humana.

OBJECTIVES: Epidural fat deposition, which varies at different levels of the vertebral column, generates a reservoir from which retained lipophilic substances could be redistributed. The aim of the study was to determine whether fat is deposited within or underneath the dural sheath or whether it is only found within the epidural space, outside the sheath. MATERIAL AND METHODS: Samples of dural sheath from the lumbar spine of human cadavers aged 65 to 72 years were analyzed by scanning electron microscopy. RESULTS: The dural sheaths were made up of an arachnoid layer and a dura mater with a thickness of 100 to 150 microm. A large number of adipocytes were observed between the layers of the dura mater as well as beneath it. CONCLUSIONS: Fat similar to that found in the peripheral nerves is present within and underneath the dural sheaths. The fat found in the dural sheaths would be in close contact with the axons of the nerve roots, unlike the fat contained in the epidural space. The release of lipophilic substances from the fat in the dural sheath could have a greater effect on the nerve roots due to the limited distance that separates the fat from the axons as well as to the poor vascular clearance.

[Epidural fat in various diseases: contribution of magnetic resonance imaging and potential implications for neuro axial anesthesia]. / La grasa epidural en diferentes patologías. Aportaciones de la resonancia magnética y posibles implicaciones en la anestesia neuroaxial.

Epidural fat is a reservoir of lipophilic substances that cushions the pulsatile movements of the dural sac, protects nerve structures, and facilitates the movement of the dural sac over the periosteum of the spinal canal during flexion and extension. Excessive epidural fat can compress the underlying structures, however, and affect the placement of catheters and the distribution of injected solutions. This review discusses changes in epidural fat related to various diseases and events: lipomatosis, epidural lymphoma, arachnoid cysts, epidural hematoma, meningiomas, angiolipomas, spondylolysis, scoliosis, spinal stenosis, and liposarcoma. Also discussed are the sequencing and protocols for magnetic resonance imaging that enable epidural fat to be observed and distinguished from neighboring structures. The relevance of epidural fat in spinal surgery is considered. Finally, we discuss the possible anesthetic implications of the abnormal deposition of epidural fat, to explain the unexpected complications that can arise during performance of epidural anesthesia.

[Bilateral axillary brachial plexus block guided by multiple nerve stimulation and ultrasound in a multiple trauma patient]. / Bloqueo axilar bilateral del plexo braquial guiado por neuroestimulación múltiple y ultrasonidos en una paciente con politraumatismo.

We present the case of a woman with multiple wounds and injuries after attempted suicide by jumping from a high place. She had multiple craniofacial injuries and fractures of both forearms requiring emergency osteosynthesis. The neurosurgeons requested that a level of consciousness be maintained for frequent assessment; therefore it was decided to provide a bilateral axillary brachial plexus block. The procedure was carried out with the aid of a nerve stimulator to locate a triple response in the left arm (radial, medial and musculocutaneous nerves) and with both ultrasound and double nerve stimulation in the right arm (medial and radial nerves). Surgery proceeded without adverse events. The location of nerves or nerve roots with both ultrasound and stimulators was highly useful in this patient in need of bilateral brachial plexus blockade. This combination, and ultrasound in particular, might be the technique of choice because it offers an image in real time and assessment of the least amount of anesthetic that seems to be needed for achieving a block.

[Possibility of cauda equina nerve root damage from lumbar punctures performed with 25-gauge Quincke and Whitacre needles]. / Posibilidad de lesión en las raíces nerviosas de la cola de caballo en relación con las punciones lumbares realizadas con agujas 25-G Quincke y Whitacre.

OBJECTIVE: To assess the possibility of puncturing nerve roots in the cauda equina with spinal needles with different point designs and to quantify the number of axons affected. MATERIAL AND METHODS: We performed in vitro punctures of human nerve roots taken from 3 fresh cadavers. Twenty punctures were performed with 25-gauge Whitacre needles and 40 with 25-gauge Quincke needles; half the Quincke needle punctures were carried out with the point perpendicular to the root and the other half with the point parallel to it. The samples were studied by optical and scanning electron microscopy. The possibility of finding the needle orifece inserted inside the nerve was assessed. On a photographic montage, we counted the number of axons during a hypothetical nerve puncture. RESULTS: Nerve roots used in this study were between 1 and 2.3 mm thick, allowing the needle to penetrate the root in the 52 samples studied. The needle orifice was never fully located inside the nerve in any of the samples. The numbers of myelinized axons affected during nerve punctures 0.2 mm deep were 95, 154, and 81 for Whitacre needles, Quincke needles with the point held perpendicular, or the same needle type held parallel, respectively. During punctures 0.5 mm deep, 472, 602, and 279 were affected for each puncture group, respectively. The differences in all cases were statistically significant. CONCLUSIONS: It is possible to achieve intraneural puncture with 25-gauge needles. However, full intraneural placement of the orifice of the needle is unlikely. In case of nerve trauma, the damage could be greater if puncture is carried out with a Quincke needle with the point inserted perpendicular to the nerve root.

Characterization of Melanoplus sanguinipes oviposition stimulating protein expression and re-examination of its potential role in stimulating oviposition.

Melanoplussanguinipes oviposition stimulating protein (MsOSP) was characterized and its role in stimulating oviposition in virgin females was examined. A 967nt MsOSP mRNA sequence with homology to previously characterized N-terminal amino acid sequence data for MsOSP was identified in a RNAseq library generated from an mRNA pool from the long hyaline tubule (LHT) of the male accessory gland complex. This transcript contained a predicted 729nt open reading frame encoding the 242aa putative MsOSP protein and had the second highest read abundance in the library. The MsOSP transcript was detected exclusively in the LHT tissue of adult males and its abundance increased with time until 7 days post-eclosion. Western blot analysis using an anti-MsOSP antibody showed high levels of MsOSP protein in the LHT luminal secretions of virgin males and to a lesser degree was associated with the aedeagus and ejaculatory duct. MsOSP was shown to be a major protein component of the spermatophore packet transferred from the male to female during copulation. However, only minor amounts of MsOSP could be detected in the female bursa, spermatheca and oviduct. Intrahemocoelic injection of LHT luminal protein into mature virgin females stimulated oviposition in ∼ 65% of females. A similar but non-significant trend was observed upon injection of purified recombinant MsOSP protein, and immunoprecipitation of LHT protein with anti-MsOSP antibody led to abrogation of oviposition stimulation upon injection of mature virgin females. Despite the demonstration of stimulation of oviposition upon intrahemocoelic injection of LHT-derived-MsOSP into mature virgin females, the potential mode of action of MsOSP in this process remains to be determined. MsOSP cannot be detected in the tissues other than the bursa, spermatheca and oviduct of female grasshoppers and relatively large quantities of MsOSP are required to stimulate oviposition upon injection.

Molecular evidence for selection on female color polymorphism in the damselfly Ischnura graellsii.

The significance of female color polymorphism in Odonata remains controversial despite many field studies. The importance of random factors (founder effects, genetic drift and migration) versus selective forces for the maintenance of this polymorphism is still discussed. In this study, we specifically test whether the female color polymorphism of Ischnura graellsii (Odonata, Coenagrionidae) is under selection in the wild. We compared the degree of genetic differentiation based on RAPD markers (assumed to be neutral) with the degree of differentiation based on color alleles. Weir and Cockerham's theta values showed a significant degree of population differentiation for both sets of loci (RAPD and color alleles) but the estimated degree of population differentiation (theta) was significantly greater for the set of RAPD loci. This result shows that some sort of selection contributes to the maintenance of similar color morph frequencies across the studied populations. Our results combined with those of previous field studies suggest that at least in some I. graellsii populations, density-dependent mechanisms might help to prevent the loss of this polymorphism but cannot explain the similarity in morph frequencies among populations.

Postmating sexual selection: allopatric evolution of sperm competition mechanisms and genital morphology in calopterygid damselflies (Insecta: Odonata).

Postmating sexual selection theory predicts that in allopatry reproductive traits diverge rapidly and that the resulting differentiation in these traits may lead to restrictions to gene flow between populations and, eventually, reproductive isolation. In this paper we explore the potential for this premise in a group of damselflies of the family Calopterygidae, in which postmating sexual mechanisms are especially well understood. Particularly, we tested if in allopatric populations the sperm competition mechanisms and genitalic traits involved in these mechanisms have indeed diverged as sexual selection theory predicts. We did so in two different steps. First, we compared the sperm competition mechanisms of two allopatric populations of Calopteryx haemorrhoidalis (one Italian population studied here and one Spanish population previously studied). Our results indicate that in both populations males are able to displace spermathecal sperm, but the mechanism used for sperm removal between both populations is strikingly different. In the Spanish population males seem to empty the spermathecae by stimulating females, whereas in the Italian population males physically remove sperm from the spermathecae. Both populations also exhibit differences in genital morphometry that explain the use of different mechanisms: the male lateral processes are narrower than the spermathecal ducts in the Italian population, which is the reverse in the Spanish population. The estimated degree of phenotypic differentiation between these populations based on the genitalic traits involved in sperm removal was much greater than the differentiation based on a set of other seven morphological variables, suggesting that strong directional postmating sexual selection is indeed the main evolutionary force behind the reproductive differentiation between the studied populations. In a second step, we examined if a similar pattern in genital morphometry emerge in allopatric populations of this and other three species of the same family (Calopteryx splendens, C. virgo and Hetaerina cruentata). Our results suggest that there is geographic variation in the sperm competition mechanisms in all four studied species. Furthermore, genitalic morphology was significantly divergent between populations within species even when different populations were using the same copulatory mechanism. These results can be explained by probable local coadaptation processes that have given rise to an ability or inability to reach and displace spermathecal sperm in different populations. This set of results provides the first direct evidence of intraspecific evolution of genitalic traits shaped by postmating sexual selection.

Genetic divergence of the seminal signal-receptor system in houseflies: the footprints of sexually antagonistic coevolution?

To understand fully the significance of cryptic female choice, we need to focus on each of those postmating processes in females which create variance in fitness among males. Earlier studies have focused almost exclusively on the proportion of a female's eggs fertilized by different males (sperm precedence). Yet, variance in male postmating reproductive success may also arise from differences in ability to stimulate female oviposition and to delay female remating. Here, we present a series of reciprocal mating experiments among genetically differentiated wild-type strains of the housefly Musca domestica. We compared the effects of male and female genotype on oviposition and remating by females. The genotype of each sex affected both female oviposition and remating rates, demonstrating that the signal-receptor system involved has indeed diverged among these strains. Further, there was a significant interaction between the effects of male and female genotype on oviposition rate. We discuss ways in which the pattern of such interactions provides insights into the coevolutionary mechanism involved. Females in our experiments generally exhibited the weakest, rather than the strongest, response to males with which they are coevolved. These results support the hypothesis that coevolution of male seminal signals and female receptors is sexually antagonistic.

[In vitro activity of new quinolones against clinical strains of Staphylococcus aureus of the wild type and with mutations characterized by gyrA, gyrB and grlA]. / Actividad in vitro de nuevas quinolonasfrente a cepas clínicas de Staphylococcus aureussilvestres y con mutaciones caracterizadasen gyrA, gyrB y grlA.

The aim of this study was to determine the in vitro activity of five quinolones against clinical strains of methicillin-susceptible and -resistant Staphylococcus aureus clinical isolates characterized at the molecular level with respect to the presence of mutations in genes coding for resistance to quinolones (grlA, gyrA and gyrB). The relationship between the mutations found and the activities of these quinolones was also analyzed. Trovafloxacin was the most active against methicillin-susceptible S. aureus and showed good activity against methicillin-resistant S. aureus, with a MIC90 of 2 mg/l. The grlA-gyrA double mutation was the most frequent (55% of the strains). Single mutation in grlA was detected only in 5% of strains; 39% of strains showed a wild-type genotype. The grlA-gyrA double mutants presented a high level of resistance against the fluoroquinolones tested except for trovafloxacin, with the MIC ranging between 0.5 and 4 mg/l. Wild-type strains were susceptible to all the fluoroquinolones tested and the single grlA mutants had a low level of quinolone resistance but were still below the breakpoint for resistance. Trovafloxacin and sparfloxacin were less affected by this mutation.