Crop production in the USA is frequently limited by a lack of pollinators.

Most of the world's crops depend on pollinators, so declines in both managed and wild bees raise concerns about food security. However, the degree to which insect pollination is actually limiting current crop production is poorly understood, as is the role of wild species (as opposed to managed honeybees) in pollinating crops, particularly in intensive production areas. We established a nationwide study to assess the extent of pollinator limitation in seven crops at 131 locations situated across major crop-producing areas of the USA. We found that five out of seven crops showed evidence of pollinator limitation. Wild bees and honeybees provided comparable amounts of pollination for most crops, even in agriculturally intensive regions. We estimated the nationwide annual production value of wild pollinators to the seven crops we studied at over $1.5 billion; the value of wild bee pollination of all pollinator-dependent crops would be much greater. Our findings show that pollinator declines could translate directly into decreased yields or production for most of the crops studied, and that wild species contribute substantially to pollination of most study crops in major crop-producing regions.

Protein levels and colony development of Africanized and European honey bees fed natural and artificial diets.

Pollen substitute diets are a valuable resource for maintaining strong and health honey bee colonies. Specific diets may be useful in one region or country and inadequate or economically unviable in others. We compared two artificial protein diets that had been formulated from locally-available ingredients in Brazil with bee bread and a non-protein sucrose diet. Groups of 100 newly-emerged, adult workers of Africanized honey bees in Brazil and European honey bees in the USA were confined in small cages and fed on one of four diets for seven days. The artificial diets included a high protein diet made of soy milk powder and albumin, and a lower protein level diet consisting of soy milk powder, brewer's yeast and rice bran. The initial protein levels in newly emerged bees were approximately 18-21 µg/µL hemolymph. After feeding on the diets for seven days, the protein levels in the hemolymph were similar among the protein diet groups (~37-49 µg/µL after seven days), although Africanized bees acquired higher protein levels, increasing 145 and 100% on diets D1 and D2, respectively, versus 83 and 60% in the European bees. All the protein diets resulted in significantly higher levels of protein than sucrose solution alone. In the field, the two pollen substitute diets were tested during periods of low pollen availability in the field in two regions of Brazil. Food consumption, population development, colony weight, and honey production were evaluated to determine the impact of the diets on colony strength parameters. The colonies fed artificial diets had a significant improvement in all parameters, while control colonies dwindled during the dearth period. We conclude that these two artificial protein diets have good potential as pollen substitutes during dearth periods and that Africanized bees more efficiently utilize artificial protein diets than do European honey bees.

Screening for diabetic retinopathy: a comparative trial of photography and scanning laser ophthalmoscopy.

AIMS: To evaluate the sensitivity and specificity of wide-field scanning laser ophthalmoscopy (WSLO) in the detection of referable diabetic eye disease, and to compare its performance with digital retinal photography. METHODS: Patients enrolled into the study underwent non-mydriatic WSLO imaging, then single- and dual-field mydriatic digital retinal photography, and examination with slit lamp biomicroscopy, the reference standard. Grading of retinopathy was performed in a masked fashion. RESULTS: A total of 380 patients (759 eyes) were recruited to the study. Technical failure rates for dilated single-field retinal photography, dual-field retinal photography and undilated WSLO were 6.3, 5.8 and 10.8%, respectively (0.005 < p < 0.02 for photography vs. WSLO). The respective indices for screening sensitivity were 82.9, 82.9 and 83.6% (p > 0.2). Specificity was 92.1, 91.1 and 89.5%, respectively (p > 0.2). CONCLUSIONS: Sensitivity and specificity for WSLO were similar to retinal photography. The technical failure rate was greater for the WSLO used in this study.

Susceptibility of Aethina tumida (Coleoptera: Nitidulidae) larvae and pupae to entomopathogenic nematodes.

In this study, we evaluated the potential use of entomopathogenic nematodes as a control for the beetle Aethina tumida Murray (Coleoptera: Nitidulidae). In particular, we conducted 1) four screening bioassays to determine nematode (seven species, 10 total strains tested) and application level effects on A. tumida larvae and pupae, 2) a generational persistence bioassay to determine whether single inoculations with nematodes would control multiple generations of A. tumida larvae in treated soil, and 3) a field bioassay to determine whether the nematodes would remain efficacious in the field. In the screening bioassays, nematode efficacy varied significantly by tested nematode and the infective juvenile (IJ) level at which they were applied. Although nematode virulence was moderate in screening bioassays 1-3 (0-68% A. tumida mortality), A. tumida mortality approached higher levels in screening bioassay 4 (nearly 100% after 39 d) that suggest suitable applicability of some of the test nematodes as field controls for A. tumida. In the generational persistence bioassay, Steinernema riobrave Cabanillas, Poinar & Raulston 7-12 strain and Heterorhabditis indica Poinar, Karunaka & David provided adequate A. tumida control for 19 wk after a single soil inoculation (76-94% mortality in A. tumida pupae). In the field bioassay, the same two nematode species also showed high virulence toward pupating A. tumida (88-100%) mortality. Our data suggest that nematode use may be an integral component of an integrated pest management scheme aimed at reducing A. tumida populations in bee colonies to tolerable levels.

The efficacy of small cell foundation as a varroa mite (Varroa destructor) control.

Due to a continuing shift toward reducing/minimizing the use of chemicals in honey bee colonies, we explored the possibility of using small cell foundation as a varroa control. Based on the number of anecdotal reports supporting small cell as an efficacious varroa control tool, we hypothesized that bee colonies housed on combs constructed on small cell foundation would have lower varroa populations and higher adult bee populations and more cm(2) brood. To summarize our results, we found that the use of small cell foundation did not significantly affect cm(2) total brood, total mites per colony, mites per brood cell, or mites per adult bee, but did affect adult bee population for two sampling months. Varroa levels were similar in all colonies throughout the study. We found no evidence that small cell foundation was beneficial with regard to varroa control under the tested conditions in Florida.

Macular oedema is not predicted by perifoveal single blot haemorrhages.

AIMS: To ascertain which perifoveal changes on digital retinal screening in diabetes predict the need for subsequent macular grid or focal laser therapy. METHODS: Between 1 January 2004 and 31 December 2005, all consecutive retinal images where any lesion was within one disc diameter of the fovea were reviewed. Patients were categorized by lesion at screening as having microaneurysm, single blot haemorrhage, multiple blot haemorrhages and exudates or circinate exudates within one disc diameter of the fovea. We compared these retinal images with the findings on slit lamp examination and the related decision for laser photocoagulation. RESULTS: Four hundred and twenty-four retinal images were identified. Of these, 52 were excluded, principally because of an interval between photography and clinic attendance of greater than 120 days, leaving 372 retinal images in the study group (313 patients). No patients with a single blot haemorrhage required immediate laser therapy at ophthalmology review compared with 13 (23%) of those with multiple blot haemorrhages and 36 (16%) of those with exudates or circinate lesions (P < 0.001). Thirty-nine patients with a single blot haemorrhage who did not require laser therapy underwent ongoing follow-up. None of these underwent laser therapy for maculopathy within the study time frame (9 months from initial screening event). CONCLUSIONS: In this study, no patients with a single blot haemorrhage within one disc diameter of the fovea on digital retinal screening required laser treatment.

Managed and Wild Bee Flower Visitors and Their Potential Contribution to Pollination Services of Low-Chill Highbush Blueberry (Vaccinium corymbosum L.; Ericales: Ericaceae).

Highbush blueberry (Vaccinium corymbosum L.; Ericales: Ericaceae) is an important crop grown throughout the eastern United States and Canada. Cross-pollination by insects greatly enhances pollination and fruit set in highbush blueberry. In Florida, low-chill cultivars that flower during the winter when most bees are dormant are used, thus, making it difficult to utilize and depend on unmanaged bees. We investigated flower visitation rates by managed and wild bees and the subsequent berry formation, berry weight, and number of seeds/berry in highbush blueberry fields in north-central Florida. Additionally, we tested three pollinator treatments: 1) pollinator-excluded flowers, 2) open-pollinated treatments that were available to managed and wild bees, and 3) flowers that were hand pollinated. Overall, we found seven native bee species that contribute to highbush blueberry pollination in Florida, but managed honey bees and bumble bees were the main flower visitors. Additionally, 14.5 times more blueberries formed in the open treatments than in the pollinator exclusion treatments, thus illustrating the economic impact bees have on blueberry pollination. Most of the wild bees observed visiting blueberry flowers were ground-nesting species that need uncultivated areas for nesting sites. Therefore, leaving field edges uncultivated and some undisturbed habitat may increase native bee numbers within blueberry farms over time.

Managed Bumble Bees (Bombus impatiens) (Hymenoptera: Apidae) Caged With Blueberry Bushes at High Density Did Not Increase Fruit Set or Fruit Weight Compared to Open Pollination.

Highbush blueberry (Vaccinium corymbosum L.) is an important crop grown throughout Florida. Currently, most blueberry growers use honey bees (Apis mellifera L.) to provide pollination services for highbush blueberries even though bumble bees (Bombus spp.) have been shown to be more efficient at pollinating blueberries on a per bee basis. In general, contribution of bumble bees to the pollination of commercial highbush blueberries in Florida is unknown. Herein, we determined if managed bumble bees could contribute to highbush blueberry pollination. There were four treatments in this study: two treatments of caged commercial bumble bee (Bombus impatiens Cresson) colonies (low and high weight hives), a treatment excluding all pollinators, and a final treatment which allowed all pollinators (managed and wild pollinators) in the area have access to the plot. All treatments were located within a highbush blueberry field containing two cultivars of blooming plants, 'Emerald' and 'Millennia', with each cage containing 16 mature blueberry plants. We gathered data on fruit set, berry weight, and number of seeds produced per berry. When pollinators were excluded, fruit set was significantly lower in both cultivars (<8%) compared to that in all of the other treatments (>58%). Berry weight was not significantly different among the treatments, and the number of seeds per berry did not show a clear response. This study emphasizes the importance of bumble bees as an effective pollinator of blueberries and the potential beneficial implications of the addition of bumble bees in commercial blueberry greenhouses or high tunnels.

In vitro and in vivo evaluation of dihydropyrimidinone C-5 amides as potent and selective alpha(1A) receptor antagonists for the treatment of benign prostatic hyperplasia.

alpha(1) Adrenergic receptors mediate both vascular and lower urinary tract tone, and alpha(1) receptor antagonists such as terazosin (1b) are used to treat both hypertension and benign prostatic hyperplasia (BPH). Recently, three different subtypes of this receptor have been identified, with the alpha(1A) receptor being most prevalent in lower urinary tract tissue. This paper explores 4-aryldihydropyrimidinones attached to an aminopropyl-4-arylpiperidine via a C-5 amide as selective alpha(1A) receptor subtype antagonists. In receptor binding assays, these types of compounds generally display K(i) values for the alpha(1a) receptor subtype <1 nM while being greater than 100-fold selective versus the alpha(1b) and alpha(1d) receptor subtypes. Many of these compounds were also evaluated in vivo and found to be more potent than terazosin in both a rat model of prostate tone and a dog model of intra-urethral pressure without significantly affecting blood pressure. While many of the compounds tested displayed poor pharmacokinetics, compound 48 was found to have adequate bioavailability (>20%) and half-life (>6 h) in both rats and dogs. Due to its selectivity for the alpha(1a) over the alpha(1b) and alpha(1d) receptors as well as its favorable pharmacokinetic profile, 48 has the potential to relieve the symptoms of BPH without eliciting effects on the cardiovascular system.

A controlled assessment of direct intraperitoneal insemination.

OBJECTIVE: To test the efficacy of direct intraperitoneal (IP) insemination and its effect on cell-mediated immunity. DESIGN: A prospective trial with each couple having one treatment (insemination) cycle and one control (timed intercourse) cycle performed in random order with the same ovulation stimulation in both cycles. SETTING: Secondary and tertiary referral fertility clinics; university teaching hospital. PATIENTS: Twenty-six infertile couples. Excluding pregnancy, only one couple did not complete the two cycles. INTERVENTIONS: Ovulation induction in both cycles. Intraperitoneal insemination in the insemination cycles. MAIN OUTCOME MEASURES: These were pregnancy rate (treatment versus control) and mixed lymphocyte response (MLR) sensitivity (before and after direct IP insemination treatment). RESULTS: There were four control and no treatment pregnancies. This was not a significant difference (odds ratio). Mixed lymphocyte responses in fertile subjects did not change during the menstrual cycle (Wilcoxon). There was no significant increase in MLR sensitivity to partner's cells after direct IP insemination treatment. CONCLUSIONS: This controlled study found no benefit from direct IP insemination in terms of pregnancies over control cycles. There was no evidence that direct IP insemination had increased cell-mediated immune response sensitivity to husband's cells.

Effect of mydriasis and different field strategies on digital image screening of diabetic eye disease.

AIMS: To assess the effects of (1) mydriasis and (2) single versus three field photography on screening for diabetic eye disease using digital photography METHOD: Slit lamp examination findings were compared to digital fundal photographs for the detection of any retinopathy and for referable retinopathy in 398 patients (794 eyes). A Topcon TRC-NW6S digital non-mydriatic fundus camera was used. Three photographic strategies were used: undilated single field, dilated single field, and dilated multiple fields. The photographs were presented in random order to one of two retinal screeners. For the single field photographs the screeners were masked to the use of mydriatics. In 13% of fundal photographs, grading was performed by both, rather than just one grader. RESULTS: Mydriasis reduced the proportion of ungradable photographs from 26% to 5% (p<0.001). Neither mydriasis nor three field photography improved the sensitivity or specificity for the detection of any retinopathy or of referable retinopathy when compared with undilated single field photography. The sensitivity and specificity for detecting referable retinopathy using undilated single field photography was 77% (95% CI 71 to 84) and 95 % (95% CI 93 to 97) respectively. Using dilated single field photography the figures were 81% (95% CI 76 to 87) and 92% (95% CI 90 to 94) respectively. Using dilated three field photography the figures were 83% (95% CI 78 to 88) and 93% (95% CI 91 to 96) respectively. Intergrader reliability for the detection of referable retinopathy in gradable photographs was excellent (Kappa values 0.86-1.00). CONCLUSIONS: Mydriasis reduces the technical failure rate. Mydriasis and the three field photography as used in this study do not increase the sensitivity or specificity of detecting diabetic retinopathy.

Glaucoma incidence in an unselected cohort of diabetic patients: is diabetes mellitus a risk factor for glaucoma? DARTS/MEMO collaboration. Diabetes Audit and Research in Tayside Study. Medicines Monitoring Unit.

AIMS: To evaluate whether diabetes mellitus is a risk factor for the development of primary open angle glaucoma or ocular hypertension (OHT). METHODS: A historical cohort study of an unselected population comprising all residents of the Tayside region of Scotland was performed using record linkage techniques followed by case note review. Ascertainment of prevalent diabetes was achieved using the Diabetes Audit and Research in Tayside Study (DARTS) validated regional diabetes register. Glaucoma and treated OHT were defined by encashment of community prescriptions and the statutory surgical procedure coding database. RESULTS: The study population comprised 6631 diabetic subjects and 166 144 non-diabetic subjects aged >40 years without glaucoma or OHT at study entry. 65 patients with diabetes and 958 without diabetes were identified as new cases of glaucoma or treated OHT during the 24 month study period, yielding a standardised morbidity ratio of 127 (95% CI, 96-158). Case note review demonstrated non-differential misclassification of prevalent glaucoma and OHT as incident disease (diabetic cohort 20%, non-diabetic cohort 24%; p=0.56) primarily as a result of non-compliance in medically treated disease. Removing misclassified cases and adjusting for age yielded an incidence of primary open angle glaucoma in diabetes of 1.1/1000 patient years (95% CI, 0.89-1. 31) compared to 0.7/1000 patient years (95% CI, 0.54-0.86) in the non-diabetic cohort; RR 1.57 (95% CI, 0.99-2.48). CONCLUSIONS: This study failed to confirm an association between diabetes mellitus and primary open angle glaucoma and ocular hypertension. A non-significant increase in diagnosed and treated disease in the diabetic population was observed, but evidence was also found that detection bias contributes to this association.

Quantitation of the 5HT1D agonists MK-462 and sumatriptan in plasma by liquid chromatography-atmospheric pressure chemical ionization mass spectrometry.

The 5HT1D agonist sumatriptan is efficacious in the treatment of migraines. MK-462 is a drug of the same class which is under development in our laboratories. Bioanalytical methods of high efficiency, specificity and sensitivity were required to support the preclinical and clinical programs. These assays were based on HPLC with tandem MS-MS detection. MK-462 and sumatriptan were extracted using an automated solid-phase extraction technique on a C2 Varian Bond-Elut cartridge. The n-diethyl analogues of MK-462 and sumatriptan were used as internal standards. The analytes were chromatographed using reversed-phase (nitrile) columns coupled via a heated nebulizer interface to an atmospheric pressure chemical ionization source. The chromatographic run times were less than 7 min. Both methods were precise, accurate and selective down to plasma concentrations of 0.5 ng/ml. The assay for MK-462 was adapted to separately monitor the unlabeled and 14C-labeled species of the drug following intravenous administration of radiolabeled material to man.

Use of LC-MS/MS to cross-validate a radioimmunoassay for the fibrinogen receptor antagonist, Aggrastat (tirofiban hydrochloride) in human plasma.

A method based on LC-MS/MS was developed for the determination of the fibrinogen-receptor antagonist Aggrastat in human plasma. The drug is isolated from plasma by liquid extraction and converted into its N-trifluoroacetyl derivative prior to analysis by HPLC with atmospheric pressure negative chemical ionization MS/MS detection. A structural analog is used as the internal standard and the lower quantifiable limit of the assay is 0.4 ng ml-1 with a relative standard deviation of 7%. This assay was used to cross-validate the existing immunoassay by analysis of plasma from patients receiving the drug. The specificity of the immunoassay was thereby confirmed.

The development and cross-validation of methods based on radioimmunoassay and LC/MS-MS for the quantification of the class III antiarrhythmic agent, MK-0499, in human plasma and urine.

An analytical method based on radioimmunoassay (RIA) has been developed for the determination of the antiarrhythmic agent, MK-0499, in plasma and urine. Owing to the potency of the drug, the specificity of this assay in human plasma could not be adequately determined using conventional RIA procedures. A highly specific procedure, based on LC/MS-MS, was developed to cross-validate the RIA. The lower quantifiable limits of the RIA and LC/MS-MS-based methods were 0.05 and 0.013 ng ml-1, respectively. Cross-validation data, compared using paired student's t-test regression analysis, showed excellent correlation between methods. The mass spectrometric assay was also used to simultaneously measure plasma concentrations of unlabeled and 14C-labeled MK-0499 following administration of the drug at high specific activity to volunteers.

Can congenital nasolacrimal-duct obstruction interfere with visual development? A cohort case control study.

PURPOSE: To ascertain if tear film disturbance in congenital nasolacrimal-duct obstruction (CNLDO) interferes with visual maturation. METHODS: Review of a cohort of children in which the occurrence and duration of CNLDO had been previously documented. Information obtained included acuity, refractive data, and presence or absence of strabismus. The incidence of abnormality in the CNLDO group was compared with that in controls, which were fellow eyes in those children with unilateral obstruction, and all children with no history of CNLDO. RESULTS: Of 4792 children, visual acuity data were obtained on 2249. There was no difference in the incidence of amblyopia in CNLDO children compared with controls (p < 0.89) nor in the watering eye compared with the fellow eye (p < 0.58). No correlation between refractive error or astigmatism and CNLDO (p < 0.26 and p < 0.32 respectively) was found. Strabismus was no more common in CNLDO than in controls (p < 0.89). CONCLUSIONS: This study found no evidence to suggest that visual maturation is adversely affected by allowing spontaneous resolution of CNLDO.

Precision large field scanning system for high numerical aperture lenses and application to femtosecond micromachining of ophthalmic materials.

A precision, large stroke (nearly 1 cm) scanning system was designed, built, and calibrated for micromachining of ophthalmic materials including hydrogels and cornea (excised and in vivo). This system comprises a flexure stage with an attached objective on stacked vertical and horizontal translation stages. This paper outlines the design process leading to our most current version including the specifications that were used in the design and the drawbacks of other methods that were previously used. Initial measurements of the current version are also given. The current flexure was measured to have a 27 Hz natural frequency with no load.