Three Halloween genes from the Varroa mite, Varroa destructor (Anderson & Trueman) and their expression during reproduction.

The ecdysteroid biosynthetic pathway involves sequential enzymatic hydroxylations by a group of enzymes collectively known as Halloween gene proteins. Complete sequences for three Halloween genes, spook (Vdspo), disembodied (Vddib) and shade (Vdshd), were identified in varroa mites and sequenced. Phylogenetic analyses of predicted amino acid sequences for Halloween orthologues showed that the acarine orthologues were distantly associated with insect and crustacean clades indicating that acarine genes had more ancestral characters. The lack of orthologues or pseudogenes for remaining genes suggests these pathway elements had not evolved in ancestral arthropods. Vdspo transcript levels were highest in gut tissues, while Vddib transcript levels were highest in ovary-lyrate organs. In contrast, Vdshd transcript levels were lower overall but present in both gut and ovary-lyrate organs. All three transcripts were present in eggs removed from gravid female mites. A brood cell invasion assay was developed for acquiring synchronously staged mites. Mites within 4 h of entering a brood cell had transcript levels of all three that were not significantly different from mites on adult bees. These analyses suggest that varroa mites may be capable of modifying 7-dehydro-cholesterol precursor and hydroxylations of other steroid precursors, but whether the mites directly produce ecdysteroid precursors and products remains undetermined.

Precocene-I inhibits juvenile hormone biosynthesis, ovarian activation, aggression and alters sterility signal production in bumble bee (Bombus terrestris) workers.

Juvenile hormone (JH) is an important regulator of development and physiology in insects. While in many insect species, including bumble bees, JH functions as gonadotropin in adults, in some highly eusocial insects its role has shifted to regulate social behavior including division of labor, dominance and aggression. Studying JH functions across social insect species is important for understanding the evolution of sociality; however, these studies have been limited because of the inability to reduce JH levels without surgically removing its glandular source, the corpora allata. Precocene is known to inhibit JH biosynthesis in several non-social insects, but has been poorly studied in social insects. Here, we tested whether precocene-I can effectively reduce JH levels in Bombus terrestris workers, and examined its effects on their physiology and behavior. Precocene-I treatment of three-worker groups decreased JH titer and ovarian activation, irrespective of the bees' dominance rank within the group, and was remedied by JH replacement therapy. Precocene-I also decreased aggressiveness and increased ester-sterility signal production; these changes were rank-dependent, and affected mainly the most reproductive and the least aggressive workers, respectively, and could not be remedied by JH replacement therapy. These results clearly confirm the role of JH as a gonadotropin and mediator of aggression in B. terrestris, and indicate that JH effects are associated with worker dominance rank. The ability to chemically reduce JH titer provides us with a non-intrusive method to probe the evolutionary changes associated with JH and the hormonal mechanisms that are associated with reproduction and behavior in social insects.

Bayesian NMR petrophysical characterization.

Identification of reservoir rock types is necessary for the exploration and recovery of oil and gas. It involves determining the petrophysical properties of rocks such as porosity and permeability which play a significant role in developing reservoir models, estimating the volumes of oil and gas reserves, and planning production methods. Nuclear magnetic resonance (NMR) technology is a fast and accurate tool for petrophysical rock characterization. The distributions of relaxation times (T2 distributions) offer valuable insights into the distribution of pore sizes in rocks, and these distributions are closely linked to important petrophysical parameters like porosity, permeability, and bound fluid volume (BFV). This work introduces a Bayesian estimation method for analyzing NMR data. The Bayesian approach uses prior knowledge of T2 distributions in the form of the prior mean and covariance. The Bayesian approach combines prior knowledge with observed data to obtain improved estimation. We use the Bayesian estimation method where prior information regarding the rock sample type, for example shale, is available. The estimators were evaluated on decay data simulated from synthesized distributions that replicate the features of experimental T2 distributions of three types of reservoir rocks. We compared the performance of the Bayesian method with two existing methods using porosity, bound fluid volume (BFV) geometric mean (T2LM) and root mean square error (RMSE) of the estimated T2 distribution as evaluation criteria. Additional experiments were carried out using experimental T2 distributions to validate the results. The performance of the Bayesian methods was also tested using mismatched priors. The experimental results illustrate that the Bayesian estimator outperforms other estimators in estimating the T2 distribution. The Bayesian method also outperforms the ILT method in estimating derived petrophysical properties except in cases where the noise level is below 0.1 and the T2 distributions are associated with short relaxation times.

Variable induction of vitellogenin genes in the varroa mite, Varroa destructor (Anderson & Trueman), by the honeybee, Apis mellifera L, host and its environment.

Transcript levels of vitellogenins (Vgs) in the varroa mite, Varroa destructor (Anderson & Trueman), were variably induced by interactions between the developing honeybee, Apis mellifera L, as a food source and the capped honeybee cell environment. Transcripts for two Vgs of varroa mites were sequenced and putative Vg protein products characterized. Sequence analysis of VdVg1 and VdVg2 proteins showed that each had greater similarity with Vg1 and Vg2 proteins from ticks, respectively, than between themselves and were grouped separately by phylogenetic analyses. This suggests there was a duplication of the ancestral acarine Vg gene prior to the divergence of the mites and ticks. Low levels of transcript were detected in immature mites, males and phoretic females. Following cell invasion by phoretic females, VdVg1 and VdVg2 transcript levels were up-regulated after cell capping to a maximum at the time of partial cocoon formation by the honeybee. During oviposition the two transcripts were differentially expressed with higher levels of VdVg2 being observed. A bioassay based on assessing the transcript levels was established. Increases in VdVg1 and VdVg2 transcripts were induced experimentally in phoretic females when they were placed inside a cell containing an early metamorphosing last instar bee but not when exposed to the metamorphosing bee alone. The variable response of Vg expression to the food source as well as environmental cues within the capped cell demonstrates that perturbation of host-parasite interactions may provide avenues to disrupt the reproductive cycle of the varroa mites and prevent varroasis.

Genomic organization and reproductive regulation of a large lipid transfer protein in the varroa mite, Varroa destructor (Anderson & Trueman).

The complete genomic region and corresponding transcript of the most abundant protein in phoretic varroa mites, Varroa destructor (Anderson & Trueman), were sequenced and have homology with acarine hemelipoglycoproteins and the large lipid transfer protein (LLTP) super family. The genomic sequence of VdLLTP included 14 introns and the mature transcript coded for a predicted polypeptide of 1575 amino acid residues. VdLLTP shared a minimum of 25% sequence identity with acarine LLTPs. Phylogenetic assessment showed VdLLTP was most closely related to Metaseiulus occidentalis vitellogenin and LLTP proteins of ticks; however, no heme binding by VdLLTP was detected. Analysis of lipids associated with VdLLTP showed that it was a carrier for free and esterified C12 -C22 fatty acids from triglycerides, diacylglycerides and monoacylglycerides. Additionally, cholesterol and ß-sitosterol were found as cholesterol esters linked to common fatty acids. Transcript levels of VdLLTP were 42 and 310 times higher in phoretic female mites when compared with males and quiescent deutonymphs, respectively. Coincident with initiation of the reproductive phase, VdLLTP transcript levels declined to a third of those in phoretic female mites. VdLLTP functions as an important lipid transporter and should provide a significant RNA interference target for assessing the control of varroa mites.

Precocious sexual signalling and mating in Anastrepha fraterculus (Diptera: Tephritidae) sterile males achieved through juvenile hormone treatment and protein supplements.

Sexual maturation of Anastrepha fraterculus is a long process. Methoprene (a mimic of juvenile hormone) considerably reduces the time for sexual maturation in males. However, in other Anastrepha species, this effect depends on protein intake at the adult stage. Here, we evaluated the mating competitiveness of sterile laboratory males and females that were treated with methoprene (either the pupal or adult stage) and were kept under different regimes of adult food, which varied in the protein source and the sugar:protein ratio. Experiments were carried out under semi-natural conditions, where laboratory flies competed over copulations with sexually mature wild flies. Sterile, methoprene-treated males that reached sexual maturity earlier (six days old), displayed the same lekking behaviour, attractiveness to females and mating competitiveness as mature wild males. This effect depended on protein intake. Diets containing sugar and hydrolyzed yeast allowed sterile males to compete with wild males (even at a low concentration of protein), while brewer´s yeast failed to do so even at a higher concentration. Sugar only fed males were unable to achieve significant numbers of copulations. Methoprene did not increase the readiness to mate of six-day-old sterile females. Long pre-copulatory periods create an additional cost to the management of fruit fly pests through the sterile insect technique (SIT). Our findings suggest that methoprene treatment will increase SIT effectiveness against A. fraterculus when coupled with a diet fortified with protein. Additionally, methoprene acts as a physiological sexing method, allowing the release of mature males and immature females and hence increasing SIT efficiency.

Comparison of methods for calculating the sound field due to a rotating monopole.

A spherical harmonic expansion for the sound field due to a rotating oscillating point source has recently been derived. This paper provides further confirmation of the expansion results by comparing it with two known numerical approaches to determining the sound field. In the advanced time approach-applicable for Mach numbers below 1-the sound at transmission time determines the field at an observation point from the distance from source to observation point at the transmission time. In the retarded time approach the field at the observation point at the observation time is determined by solving for the retarded transmission times. The results from all three approaches are shown to be in good agreement. Expressions for the far-field instantaneous frequency are also derived and shown to agree with previous work.

Evaluating light attraction to increase trap efficiency for Tribolium castaneum (Coleoptera: Tenebrionidae).

The red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), is a major coleopteran pest in flour mills and storage facilities. An aggregation pheromone has been identified for this pest; however, the pheromone is of limited value for population monitoring. To develop more efficient methods to monitor this pest, experiments were conducted to determine whether light functioned as an attractant for the red flour beetle. Light-emitting diodes (LEDs) of various wavelengths were examined as light sources because they produce bright, narrow light spectra. A comparison of responses to light spectra across the visible and UV regions of the electromagnetic spectrum indicated that the beetle was most attracted to near UV LED at a 390 nm dominant wavelength. The use of LEDs in competitive laboratory experiments resulted in a 20% capture of released beetles, compared with a 1% capture with the aggregation pheromone alone. Even more beetles were captured with a combination of LEDs and commercially available chemical lures in traps. LEDs can easily be added onto existing trap designs or new traps can be designed to take full advantage of positive phototaxis.

Effects of Focal Ionizing Radiation of the Squid Stellate Ganglion on Synaptic and Axonal Transmission in the Giant-Fiber Pathway.

Ionizing radiation is clinically used to treat neurological problems and reduce pathological levels of neural activity in the brain, but its cellular-level mechanisms are not well understood. Although spontaneous and stimulated synaptic activity has been produced in rodents by clinically and environmentally relevant doses of radiation, the effects on basic excitability properties of neurons have seldom been reported. This study examined the effects of focused ionizing radiation on synaptic transmission and action potential generation in the squid giant-fiber system, which includes the giant synapse between a secondary interneuron and the tertiary giant motor axons. Radiation of 140-300 Gy was delivered to a stellate ganglion of a living squid over several minutes, with the contralateral stellate ganglion serving as an internal control. No qualitative changes in the efficacy of synaptic transmission were noted in conjunction with stimulation of the input to the giant synapse, although in one irradiated ganglion, the refractory period increased from about 5 ms to more than 45 seconds. Small but significant changes in the action potential recorded from the giant motor axon in response to electrical stimulation were associated with an increased maximum rate of fall and a shortened action potential duration. Other action-potential parameters, including resting potential, overshoot, the maximum rate of the rise, and the refractory period were not significantly changed. Attempts to account for the observed changes in the action potential were carried through a Hodgkin-Huxley model of the action potential. This approach suggests that an increase in the maximum voltage-gated potassium conductance of about 50% mimics the action potential shortening and increased rate of fall that was experimentally observed. We propose that such an effect could result from phosphorylation of squid potassium channels.

Rationale and design of INTERSTROKE: a global case-control study of risk factors for stroke.

UNLABELLED: Stroke is a major global health problem. It is the third leading cause of death and the leading cause of adult disability. INTERHEART, a global case-control study of acute myocardial infarction in 52 countries (29,972 participants), identified nine modifiable risk factors that accounted for >90% of population-attributable risk. However, traditional risk factors (e.g. hypertension, cholesterol) appear to exert contrasting risks for stroke compared with coronary heart disease, and the etiology of stroke is far more heterogeneous. In addition, our knowledge of risk factors for stroke in low-income countries is inadequate, where a very large burden of stroke occurs. Accordingly, a similar epidemiological study is required for stroke, to inform effective population-based strategies to reduce the risk of stroke. METHODS: INTERSTROKE is an international, multicenter case-control study. Cases are patients with a first stroke within 72 h of hospital presentation in whom CT or MRI is performed. Proxy respondents are used for cases unable to communicate. Etiological and topographical stroke subtype is documented for all cases. Controls are hospital- and community-based, matched for gender, ethnicity and age (+/-5 years). A questionnaire (cases and controls) is used to acquire information on known and proposed risk factors for stroke. Cardiovascular (e.g. blood pressure) and anthropometric (e.g. waist-to-hip ratio) measurements are obtained at the time of interview. Nonfasting blood samples and random urine samples are obtained from cases and controls. Study Significance: An effective global strategy to reduce the risk of stroke mandates systematic measurement of the contribution of the major vascular risk factors within defined ethnic groups and geographical locations.

Myocardial infarction following t-PA for acute stroke.

BACKGROUND AND PURPOSE: Complications of intravenous (IV) thrombolysis with tissue plasminogen activator (t-PA) for acute stroke are commonly related to hemorrhage, anaphylaxis, or arterial re-occlusion. Embolic complications of t-PA are beginning to be recognized with increased use of t-PA for acute ischemic stroke. We hypothesize that disruption of intra-cardiac thrombus may result in myocardial infarction (MI) after use of t-PA for acute ischemic stroke. SUMMARY OF CASES: We describe three cases of acute MI immediately following IV t-PA infusion for acute stroke. In patient #1 apical thrombus was visualized on cardiac echocardiogram accounting for the MI after t-PA for acute stroke. Patient #2 had fresh thrombus seen on cardiac catherization after use of t-PA for acute stroke. Patient #3 developed a significant troponin rise 15 hours after the t-PA for stroke infusion with an echocardiogram revealing new wall motion abnormalities. Patient # 1 and #2 died secondary to multi-organ failure. DISCUSSION: Acute MI immediately following t-PA treatment for stroke is a rare but serious complication. The disruption of intra-cardiac thrombus and subsequent embolization to the coronary arteries may be an important mechanism in the development of MI after t-PA treatment for acute ischemic stroke.

Uveitis associated with Primary Angiitis of the Central Nervous System.

BACKGROUND: In Primary Angiitis of the Central Nervous System (PACNS), disease is typically limited to the brain and spinal cord although other organs may be affected. Uveitis is occasionally seen in systemic vasculitides but is not a recognized manifestation of PACNS. We describe two patients who developed PACNS following the onset of uveitis. CASE DESCRIPTIONS: Case 1--a 47-year-old male suffered multiple TIAs and left pontine stroke shortly after two episodes of diffuse uveitis. A cerbral angiogram demonstrated multiple caliber changes within several intracranial vessels. Cyclophosphamide was added after his stroke occurred during pulse methylprednisolone therapy. Case 2--a 35-year-old male suffered a spinal cord TIA followed by hemispheric and brainstem infarctions two months after an episode of uveitis and Bell's palsy treated with oral prednisone. A cerebral angiogram demonstrated multiple caliber changes within several intracranial vessels. He was successfully treated with oral prednisone and cyclophosphamide. CONCLUSIONS: Uveitis should be considered a recognized feature of PACNS. Combination immunosuppressive therapy with prednisone and cyclophosphamide may be necessary for successful treatment.

Male sexual enhancement after methoprene treatment in Anastrepha fraterculus (Diptera: Tephritidae): A sustained response that does not fade away after sexual maturation.

The juvenile hormone (JH) of insects triggers physiological changes related to reproduction in adults of both sexes. Methoprene is a sesquiterpene with some effects that are analogous to those of JH. Treatments with methoprene accelerate sexual maturation in males of the South American fruit fly Anastrepha fraterculus, giving young males a mating advantage over non-treated males of the same age. Here, we evaluated the effects of methoprene treatment on A. fraterculus males after the sexual maturation phase and tested whether this compound provides a long-term mating advantage. Moreover, we took the first step to unravel the mechanisms that underlie male sexual enhancement. We treated males 1day or 8days after adult emergence and compared mate choice between recently matured (young) females and females that had been mature for ca. 10days (aged females). We also addressed methoprene treatment effects on male sexual signalling. We found that methoprene treatment enhanced male sexual competitiveness even after the sexual maturation phase, and the effect did not decrease until males were older than 20days. However, when methoprene treatment was carried out close to sexual maturity, the mating enhancement was no longer observed, suggesting a non-immediate effect and excluding the possibility that methoprene acts as a pheromonal compound. Young and aged females tended to mate more frequently with treated-males. This might indicate that in a context of sexual selection, the potential benefits associated with reproductive success would be similar for females of both ages. Treated males released larger amounts of pheromonal compounds than non-treated males, but their courtship behaviour was not altered to the same extent, suggesting that methoprene treatment may accelerate differently the components of male courtship. We discuss potential benefits of using methoprene to increase the efficiency of the sterile insect technique, which is an environmentally safe method to control this important South American fruit pest.

Reduction in IV t-PA door to needle times using an Acute Stroke Triage Pathway.

OBJECTIVE: To determine the effectiveness of an Acute Stroke Triage Pathway in reducing door to needle times in acute stroke treatment with IV t-PA. BACKGROUND: A previous study at our tertiary referral centre, examining IV t-PA door to needle times, was completed in 2000. The median door to needle time was beyond the recommended National Institute for Neurological Disorders and Stroke (NINDS) standard of 60 minutes. In November 2001, an Acute Stroke Triage Pathway was introduced in the emergency room (ER) to address this issue. The goal of this pathway was to rapidly identify patients eligible for treatment for IV t-PA, so that CT scans and lab studies could be arranged immediately upon ER arrival. Our hypothesis was that the Triage Pathway would shorten door to CT and door to needle times. DESIGN/METHODS: Using retrospective data, pre (n=87) and post (n=47) triage pathway times were compared. The door to CT time was reduced by 11 minutes (p=0.015) and door to needle time was reduced by 18 minutes (p=0.0036) in a subgroup of patients that presented directly to our hospital. CONCLUSIONS: These results indicate that the Acute Stroke Triage Pathway is effective in reducing Door to CT and Door to Needle Times in patients presenting directly to our ER. However, a majority of treatment times were still beyond NINDS recommendations. Stroke Centers require periodic review of their efficiency to ensure that target times are being obtained and may benefit from the use of an Acute Stroke Triage Pathway.

Comparison of Host-Seeking Behavior of the Filth Fly Pupal Parasitoids, Spalangia cameroni and Muscidifurax raptor (Hymenoptera: Pteromalidae).

The pupal parasitoids, Spalangia cameroni Perkins and Muscidifurax raptor Girault and Sanders, can be purchased for biological control of house flies Musca domestica L. and stable flies Stomoxys calcitrans (L.) (Diptera: Muscidae). Little is known about the odors involved in host-seeking behavior of these two species, so odors associated with house flies were investigated in the laboratory using a Y-tube olfactometer. Odor stimuli from house fly host puparia, larvae, pine-shavings bedding with horse manure, and developing flies in the pine-shavings-manure substrate were evaluated in bioassays using the two pteromalid species. In choice tests, naïve female S. cameroni were strongly attracted to odor from the substrate containing house fly larvae and secondarily from the uninfested substrate and substrate with puparia versus humidified and purified air. This species also selected the substrate with larvae versus the substrate with the house fly puparia or uninfested substrate. Muscidifurax raptor was attracted to odor from the substrate containing puparia, washed puparia, and substrate with puparia removed. The data suggest that coexistence between the two pteromalid parasitoids, S. cameroni and M. raptor, might be promoted by different host-seeking behavior.

Intravenous tissue plasminogen activator for acute ischemic stroke: A Canadian hospital's experience.

BACKGROUND AND PURPOSE: In the United States, tissue plasminogen activator (tPA) was approved for treatment of acute ischemic stroke in 1996. Its use has only recently been approved in Canada. We sought to evaluate the safety, feasibility, and efficacy of treatment in a Canadian hospital setting. METHODS: A combined retrospective and prospective review is presented of 46 consecutive patients treated with intravenous tPA at our hospital with a treatment protocol similar to that of the National Institute of Neurological Disorders and Stroke (NINDS) trial. RESULTS: Symptomatic intracranial hemorrhage at 36 hours occurred in 1 patient (2.2%). The median time to treat was 165 minutes, with a median "door-to-needle" time of 84 minutes. Compared with patients presenting initially at our hospital, patients transferred from another institution for tPA therapy were treated closer to the 3-hour time window (mean 173 versus 148 minutes, P:<0.001) but had a shorter door-to-needle time (43 versus 102 minutes, P:<0.001). For every 10 minutes closer to the 3-hour time window that any patient arrived at the hospital, 7 minutes was saved in the door-to-needle time (correlation coefficient 0.9, P:<0.001). Patient outcome did not differ from that in the NINDS trial (P:>0.75). CONCLUSIONS: Our safety and patient outcome data compare favorably with NINDS and Phase IV data. Although a 3-hour treatment window was feasible, the median door-to-needle time lengthened as more treatment time was available and the door-to-needle time was beyond recommended standards. This review has prompted changes in our community to improve treatment efficiency.

Cerebellar infarcts in the New England Medical Center Posterior Circulation Stroke Registry.

We report the clinical findings and stroke mechanisms of 63 patients with cerebellar infarcts. We divided the intracranial vertebrobasilar circulation into the proximal territory (P), fed by the intracranial vertebral arteries and their branches; the middle territory (M), fed by the proximal and middle basilar artery and its branches; and the distal territory (D), fed by the rostral basilar artery and its branches. Cerebellar infarcts were classified by vascular territories P, M, D, P&D, and middle-plus (P&M, M&D, and P&M&D). Patients with P infarcts (11 patients) frequently had vertigo, gait instability, limb ataxia, and headache, whereas patients with D infarcts (15 patients) most often had limb ataxia, gait instability, and dysarthria. Patients with P&D infarcts (17 patients) had signs and symptoms of both groups combined. Infarcts in which the middle territory was involved, either alone (three patients) or combined with other territories (17 patients) were dominated by brainstem signs and symptoms. The predominant stroke mechanisms in the P, D, and P&D groups were embolic due to intra-arterial or cardiac embolism. When the M territory was involved, either alone or with P, D, or P&D territories, stroke mechanisms were more varied, and there was often large-artery occlusion with hemodynamic ischemia.

Intermittent downbeat nystagmus due to vertebral artery compression.

Downbeat nystagmus (DBN) uncommonly occurs as a transient phenomenon, and it rarely occurs in patients with cerebrovascular disease. We observed a patient with intermittent DBN and lightheadedness due to transient obstruction of his dominant vertebral artery when he turned his head to his left side. Surgical removal of an osteophyte at the site of the angiographically demonstrated lesion relieved his symptoms.

Embolism from vertebral artery origin occlusive disease.

We report 10 patients with severe occlusive disease of the vertebral artery (VA) origin in the neck with intra-arterial embolism to the posterior circulation. The VA lesions in seven patients were complete occlusions, and three patients had severe atherostenosis. All patients had strokes in the vertebrobasilar territory. The most frequent recipient sites of intra-arterial embolism were the intracranial VA-posterior inferior cerebellar artery region (8), and the distal basilar artery (BA) and its superior cerebellar and posterior cerebral artery branches (7). Two patients had pontine infarction due to BA embolism. The most common clinical signs were due to cerebellar infarction. Atherosclerotic disease of the VA origin has features in common with disease of the internal carotid artery origin. Both have similar risk factors and demography, and each can cause strokes by intracranial intra-arterial embolism.

Clozapine in the treatment of parkinsonian patients with dopaminomimetic psychosis.

In a double-blind placebo-controlled study, we evaluated the effects of clozapine (75 to 250 mg/day, mean 170.8) on dopaminomimetic psychosis and parkinsonian disability. Clozapine prevented deterioration of psychosis during the increase of dopaminomimetics in the 3 patients who completed the study. Worsening of parkinsonism occurred in 3 of the 6 patients. In the dosage used, clozapine's usefulness was limited by its propensity to produce sedation, confusion, and increased parkinsonism.