Indigenous peoples and local communities as partners in the sequencing of global eukaryotic biodiversity.

The aim to sequence, catalog, and characterize the genomes of all of Earth's eukaryotic biodiversity is the shared mission of many ongoing large-scale biodiversity genomics initiatives. Reference genomes of global flora and fauna have the potential to inform a broad range of major issues facing both biodiversity and humanity, such as the impact of climate change, the conservation of endangered species and ecosystems, public health crises, and the preservation and enhancement of ecosystem services. Biodiversity is dramatically declining: 28% of species being assessed by the IUCN are threatened with extinction, and recent reports suggest that a transformative change is needed to conserve and protect what remains. To provide a collective and global genomic response to the biodiversity crisis, many biodiversity genomics initiatives have come together, creating a network of networks under the Earth BioGenome Project. This network seeks to expedite the creation of an openly available, "public good" encyclopedia of high-quality eukaryotic reference genomes, in the hope that by advancing our basic understanding of nature, it can lead to the transformational scientific developments needed to conserve and protect global biodiversity. Key to completing this ambitious encyclopedia of reference genomes, is the ability to responsibly, ethically, legally, and equitably access and use samples from all of the eukaryotic species across the planet, including those that are under the custodianship of Indigenous Peoples and Local Communities. Here, the biodiversity genomics community is subject to the provisions codified in international, national, and local legislations and customary community norms, principles, and protocols. We propose a framework to support biodiversity genomic researchers, projects, and initiatives in building trustworthy and sustainable partnerships with communities, providing minimum recommendations on how to access, utilize, preserve, handle, share, analyze, and communicate samples, genomics data, and associated Traditional Knowledge obtained from, and in partnership with, Indigenous Peoples and Local Communities across the data-lifecycle.

Fitting acrylic occlusal splints and an experimental laminated appliance used in migraine prevention therapy.

OBJECTIVE: To evaluate clinical procedures and chair time required to seat and adjust hard, heat-cured acrylic occlusal splints and an alternative laminated appliance developed to simplify construction of migraine prevention appliances. DESIGN AND SETTING: Single-centre study in the Oral Medicine Clinic, The Royal Hospitals, Belfast, Northern Ireland. METHOD: Questionnaires were distributed, January-May 2003, to operators fitting occlusal splints for 100 consecutive patients selected for migraine prevention therapy. Half the appliances were made in heat-polymerised acrylic with the remainder using a novel combination of ethylene vinyl acetate and light-curing urethane dimethacrylate. Information on operator experience, the nature of any fitting surface and occlusal adjustments together with an estimate of the time taken to make alterations was recorded. KEY FINDINGS: The need for adjustment to seat appliances intraorally was significantly less for migraine prevention appliances made using an experimental laminating technique. Where modifications were necessary, there was no significant difference in the chair time required to fit either the heat-cured hard or experimental laminated migraine prevention appliance. CONCLUSION: Provision of migraine prevention appliances may be more time efficient if the dental practitioner considers a laminated approach to construction.

Cholinergic mechanisms in canine narcolepsy--I. Modulation of cataplexy via local drug administration into the pontine reticular formation.

Cataplexy in the narcoleptic canine has been shown to increase after systemic administration of cholinergic agonists. Furthermore, the number of cholinergic receptors in the pontine reticular formation of narcoleptic canines is significantly elevated. In the present study we have investigated the effects of cholinergic drugs administered directly into the pontine reticular formation on cataplexy, as defined by brief episodes of hypotonia induced by emotions, in narcoleptic canines. Carbachol and atropine were perfused through microdialysis probes implanted bilaterally in the pontine reticular formation of freely moving, narcoleptic and control Doberman pinschers. Cataplexy was quantified using the Food-Elicited Cataplexy Test, and analysed using recordings of electroencephalogram, electrooculogram and electromyogram. Cataplexy was characterized by a desynchronized electroencephalogram and a drop in electromyogram and electrooculogram activity. In narcoleptic canines, both unilateral and bilateral carbachol (10(-5) to 10(-3) M) produced a dose-dependent increase in cataplexy, which resulted in complete muscle tone suppression at the highest concentration. In control canines, neither bilateral nor unilateral carbachol (10(-5) to 10(-3) M) produced cataplexy, although bilateral carbachol, did produce muscle atonia at the highest dose (10(-3)). The increase in cataplexy after bilateral carbachol (10(-4) M) was rapidly reversed when the perfusion medium was switched to one containing atropine (10(-4) M). Bilateral atropine (10(-3) to 10(-2) M) alone did not produce any significant effects on cataplexy in narcoleptic canines; however, bilateral atropine (10(-2) M) did reduce the increase in cataplexy produced by systemic administration of physostigmine (0.05 mg/kg, i.v.). These findings demonstrate that cataplexy in narcoleptic canines can be stimulated by applying cholinergic agonists directly into the pontine reticular formation. The ability of atropine to inhibit locally and systemically stimulated cataplexy indicates that the pontine reticular formation is a critical component in cholinergic stimulation of cataplexy. Therefore, it is suggested that the pontine reticular formation plays a significant role in the cholinergic regulation of narcolepsy.

Limb-threatening ischemia in the medically compromised patient: amputation or revascularization?

The clinical courses of 362 patients with threatened limbs, seen over a 25-year period, were reviewed. Below-knee amputation was elected in 158 patients and infrainguinal revascularization in 204 patients. Patients were excluded from the study if amputation was chosen on the basis of arteriographic findings, diffuse infection, or extensive tissue loss. In an effort to compare the two treatment modalities in a minimally biased fashion, patients were stratified into three classes on the basis of the Goldman Multifactorial Index of Cardiac Risk and the American Society of Anesthesiology classification. Within each class, patients treated with amputation and with bypass were comparable with respect to age, sex, severity of ischemia, and anesthetic technique. Patients who underwent revascularization had a lower perioperative mortality rate (p less than 0.05), a shorter length of hospital stay (p less than 0.05), and an increased long-term survival rate (p less than 0.05) than the medically matched subgroup of patients who underwent amputation. Patients in the revascularization group were more successful in regaining ambulatory abilities than patients in the amputation group (p less than 0.01). These differences were most significant as the degree of medical compromise increased. Although medically compromised patients have frequently been denied revascularization on the basis of presumed increases in surgical risk and decreased long-term survival, the data appear to suggest that it is precisely the patients of this group who appear to benefit the most from attempts at limb salvage.

Determination of ML-1035 enantiomers in plasma by chiral high-performance liquid chromatography.

ML-1035, is a gastroprokinetic agent structurally related to metoclopramide. Because ML-1035 contains an asymmetric chiral sulphoxide moiety, a chiral HPLC method has been developed to separate and quantitate its R- and S-enantiomers in plasma. The ML-1035 enantiomers present in plasma are extracted with dichloroethane under alkaline conditions, the extract evaporated to dryness and reconstituted in the mobile phase. Samples are chromatographed on a Chiralcel OD HPLC column with hexane-absolute ethanol (1% TEA) (1:1, v/v) as the mobile phase. The enantiomers of the unchanged drug are determined by fluorescence measurement (ex: 310 nm, em: 350 nm). The method provides a linear response for both enantiomers over a concentration range of 25 (limit of determination) to 2500 ng ml-1 with correlation coefficients of 0.9987 or greater. The inter-assay precision is 9.5% or less and the accuracy ranges from 93.9 to 103.4% of the theoretical value. The method is used to determine the plasma concentrations of the R- and S-enantiomers following oral and intravenous administration of R- or S-enantiomers to dogs. The method is also adapted to measure enantiomer levels from in vitro reaction mixtures so that the possibility of metabolic inversion may be assessed. The data suggest that no significant level of inversion between the enantiomers occurred either in vivo or in vitro.

Revascularization of the lower extremity with omentum.

The use of the omental graft with multiple level arterial anastomoses may prove useful for salvaging the severely ischemic lower limb. Because the long-term results of full free omental grafts may depend on neovascularization of the omentum from the surrounding tissue, the technique described here should be utilized only in those patients who are not suitable candidates for other procedures.

Colour perception of laboratory-fired samples of body-coloured ceramic.

The objective of this investigation is to test by common perception, the colour fidelity of laboratory-fired samples of ceramic. Thirty-two ceramic discs were constructed in a thickness similar to that used for ceramic veneers using eight shades from each of four different manufacturers. A group of 20 'adept' observers colour matched the samples in strictly controlled viewing conditions using a Vita Lumin (Vita Zahnfabrik) shade guide. Samples were categorised as 'positive matches'(matching the expected shade), 'agreed shades' (matching an unexpected shade), or as 'unidentified shades' (no match). The Vita Lumin (Vita Zahnfabrik) colour 'value' index was used as a reference to categorise colour 'value' differences between observed and expected shade. Colour 'value' selection was divided into three groups: (1) matching colour 'value', (2) shades of higher colour 'value' than expected or (3) shades of lower colour 'value' than expected. Only six of the 32 samples examined were perceived to be 'positive matches' with the recommended shade guide. Fourteen 'agreed shades' were found while the remaining 12 shades were regarded as 'unidentified shades'. The vast majority of observations disagreed with the manufacturers' designated shade. A significant trend was found favouring higher 'value' (lighter) shades than those specified by the manufactures. Significant differences in the pattern of shade matches and disagreements between brands were also found. Alternative shade determination systems and/or ceramic colour modifications are required if colour fidelity between the shade guide and the materials tested is to be improved.

Lymphatic drainage from the groin following surgery of the femoral artery.

Lymphatic drainage from the surgical wound is uncommon after most surgical procedures. Little data regarding this problem is encountered in the American literature. The incidence of this complication is not established. Data are not available regarding speculative risk factors, including diabetes reoperative dissection, infection, and use of prosthetic materials. The likelihood secondary infection is not established. A review of 428 wounds following dissection of the femoral artery was undertaken, with analysis of resulting wound drainage. Lymphatic drainage occurred in 5% of groin wounds. Diabetes, infection of the operated leg or foot and use of prosthetic graft were associated with statistically insignificant increases in risk. Reoperation was associated with a statistically insignificant decrease in risk. Most drainage stopped spontaneously. In 40% of patients with drainage, protracted lymphorrhea contributing to morbidity and hospital stay resulted. This outcome was unlikely in the absence of prosthetic graft. Such patients may benefit from surgical exploration of the wound and ligation of lymphatic tissue. Graft infection was not observed. While infection was an infrequent secondary complication, bacteriologic monitoring and antibiotic therapy are essential in cases of prolonged drainage.

Vocal cord paralysis following carotid endarterectomy.

From 1964 to 1977, over 400 carotid endarterectomies were performed at the Rochester General Hospital. Two patients from this group experienced hoarseness secondary to vocal cord paralysis on the operated side, believed due to trauma to the vagus nerve. Although uncommon, it is important to stress the method of avoiding this complication.

Femoro-popliteal by-pass with bovine heterografts.

Modified bovine heterografts were used in 28 patients, primarily extremity salvage problems, found to have unavailable or unusable saphenous veins. The accumulated 3 year patency rate was 35%. Of the 18 graft failures, 11 required major amputation. A high failure rate of 39% in the first six-month interval was observed. Electromagnetic flowmeter recording were of limited value in predicting patency. Continued evaluation of bovine grafts for femoro-popliteal reconstruction in the absence of saphenous vein is recommended.

Acute aortic rupture from cross clamp injury.

We report a case of transmural disruption and perforation of the abdominal aorta secondary to clamp injury. The literature is reviewed in reference to the traumatic effects of vascular clamps. Steps are outlined to reduce the risk of this injury.

A laboratory system for production of flexion rates and forces in the forelimb of the horse.

The distal portion of the forelimb of the horse is provided with a stay apparatus composed of tendons, ligaments, and fascia. This stay apparatus provides the major resistance to joint flexion during the support phase of the stride. The laboratory test system described was shown to be able to reproduce in vitro limb motions and hoof forces measured with a running horse. These results indicated the stay apparatus operates in a largely passive mode, active muscle contraction apparently serving to provide rigidity only early in the support phase of the stride. The testing system described was designed to take advantage of the passive nature of the stay apparatus in order to study in vitro the forces, accelerations, and deformations encountered by the limb of the galloping horse in vivo.

Equine degenerative myeloencephalopathy.

Five horses of mixed breeding and 1 zebra with symmetric tetraparesis and ataxia were studied. There were 2 females and 4 males. Signs of illness were first noticed from birth to 12 months of age. Clinical and pathologic studies were performed when the animals were 4 to 24 months old. Two of the horses were related. All of the subjects had diffuse degenerative myeloencephalopathy. The degeneration was most pronounced in thoracic segments of the spinal cord. Ventral and dorsolateral funiculi were most severely affected. Axonal swelling and abnormal neuronal cell bodies were seen in the gray matter of the spinal cord and in some brain-stem nuclei. These changes were most prominent in the nuclei of the dorsal spinocerebellar tract. The cause was not determined.

Use of a common shade guide to test the perception of differences in the shades and value by members of the dental team.

PURPOSE: Commercial shade guides are frequently used for treatments where the shades of teeth are being reproduced or altered. Where differences between shade tabs and the natural teeth are small, difficulty is often experienced in determining the closest match. The purpose of this investigation is to discover whether the claimed colour differences between individual shades of one commonly used guide can be determined subjectively in clinical circumstances. BASIC PROCEDURES: Seventy dental team professionals made shade comparisons in an idealized colour-matching environment using shade guides as supplied by the manufacturer. Their arrangements by 'value' were recorded and unidentified shade tabs were matched with those on a complete guide. MAIN FINDINGS: The results showed that discrepancies existed between nominated and perceived colours, for the shade determination system tested. The shade-guide manufacturer's recommended arrangement of shade tabs ranked according to colour 'value' did not correspond to subjective perception of 'value' ordering under the conditions of the study. PRINCIPAL CONCLUSIONS: There may be no practical clinical advantage in having shades which cannot be distinguished. This study indicates that colour differences between shade tabs are not clearly perceived by members of the dental team. A careful reduction of the number of tabs in the guide tested might simplify shade selection procedures and help to standardise shade-taking.

Identification of Dirofilaria immitis proteins recognized by antibodies from infected dogs.

The identification of excreted-secreted (ES) proteins of filarial nematodes as potential diagnostic reagents is an important requirement for the development of methods to determine level of infection in the host, especially for human filariae. Dirofilaria immitis, the canine heartworm, is a widespread and important veterinary pathogen and is a useful model for filarial parasites of humans. An analysis of proteins released from adult D. immitis (the secretome) in culture is available. We sought to identify D. immitis ES proteins found in vivo to validate the in vitro secretome and to investigate them as potential diagnostic reagents. Cultures of D. immitis adults obtained from infected dogs were maintained for 72 hr with daily changes of media. Proteins were concentrated from spent media by standard methods and were passed through Protein-A columns containing purified IgG antibodies from heartworm-infected dogs. Following extensive washing, heartworm proteins recognized by the antibodies were eluted from these columns and submitted for analysis by tandem mass spectrometry (MS/MS). As a comparison, somatic proteins from adult D. immitis female parasites and microfilaria were also processed and analyzed by the same protocol. Six, 9, and 12 proteins were identified by MS/MS in the ES, adult female, and microfilaria samples, respectively. The identification of the most abundant parasite proteins present in the serum of infected hosts offers a rational approach to the development of new diagnostic assays that may be applicable across the Filarioidea.