Convergent evolution of pain-inducing defensive venom components in spitting cobras.

Convergent evolution provides insights into the selective drivers underlying evolutionary change. Snake venoms, with a direct genetic basis and clearly defined functional phenotype, provide a model system for exploring the repeated evolution of adaptations. While snakes use venom primarily for predation, and venom composition often reflects diet specificity, three lineages of cobras have independently evolved the ability to spit venom at adversaries. Using gene, protein, and functional analyses, we show that the three spitting lineages possess venoms characterized by an up-regulation of phospholipase A2 (PLA2) toxins, which potentiate the action of preexisting venom cytotoxins to activate mammalian sensory neurons and cause enhanced pain. These repeated independent changes provide a fascinating example of convergent evolution across multiple phenotypic levels driven by selection for defense.

Transcriptomic meta-signatures identified in Anopheles gambiae populations reveal previously undetected insecticide resistance mechanisms.

Increasing insecticide resistance in malaria-transmitting vectors represents a public health threat, but underlying mechanisms are poorly understood. Here, a data integration approach is used to analyse transcriptomic data from comparisons of insecticide resistant and susceptible Anopheles populations from disparate geographical regions across the African continent. An unbiased, integrated analysis of this data confirms previously described resistance candidates but also identifies multiple novel genes involving alternative resistance mechanisms, including sequestration, and transcription factors regulating multiple downstream effector genes, which are validated by gene silencing. The integrated datasets can be interrogated with a bespoke Shiny R script, deployed as an interactive web-based application, that maps the expression of resistance candidates and identifies co-regulated transcripts that may give clues to the function of novel resistance-associated genes.

Isolation and characterization of renin-like aspartic-proteases from Echis ocellatus venom.

Three aspartic proteases (SVAPs) have been isolated from venom of the saw-scaled viper, Echis ocellatus. In confirmation of prior transcriptomic predictions, all three forms match to sequences of either of the two SVAP transcripts (EOC00051 and EOC00123), have a molecular weight of 42 kDa and possess a single N-glycan. The SVAPs act in a renin-like manner, specifically cleaving human and porcine angiotensinogen into angiotensin-1 and possess no general protease activity. Their activity is completely inhibited by the aspartyl protease inhibitor Pepstatin A.

Worldwide host plants of the highly polyphagous, invasive Epiphyas postvittana (Lepidoptera: Tortricidae).

The light brown apple moth, Epiphyas postvittana (Walker) (Lepidoptera: Tortricidae), is a highly successful biological invader. It was accidentally introduced to several countries including New Zealand, Hawaii, England, and California. Light brown apple moth attacks a wide range of crop plants and other woody and herbaceous plants, but a more comprehensive analysis of its host range is needed for risk assessments, to evaluate the likely economic and environmental impacts, and to enable targeting of particular plant species for detection surveys and treatments. We reviewed and synthesized the host range and host selection behavior of light brown apple moth by using information from Australia and invaded countries. The host range of light brown apple moth is determined by the behavior of both adult females and larvae. Females use visual, chemical and physical cues to choose host plants. Larvae are capable of limited active dispersal by walking and longer range dispersal by ballooning on silken strands; therefore, larvae also may need to select host plants. We review larval performance indicators across a range of plants. Based on our review, there are at least 545 plant species in 363 genera from 121 families that have been reported as hosts of light brown apple moth. Some plants were reported only once and need verification. Nevertheless, many host plant species and their wide phylogenetic range (from ferns to higher dicotyledons) indicates that light brown apple moth is one of the most polyphagous insects known. This information and our categorization of frequency of host use are valuable for incursion response and pest management activities.

Classification of cochlear implant failures and explantation: the Melbourne experience, 1982-2006.

The definition and categorization of reasons for cochlear implant (CI) failure have recently been standardized following the publication of the European Consensus Statement on Cochlear Implant Failures and Explantation (ECSCIFE) (2005). The aim of this study was to review the Melbourne experience with cochlear implant failure and reimplantation, applying ECSCIFE guidelines for categorization and assessing hearing outcomes. A retrospective review was carried out of Melbourne CI clinic's records to identify all cases of implant failure and categorize them using ECSCIFE guidelines. Comparison was made of pre- and post-reimplantation hearing levels for those patients who had undergone ipsilateral reimplantation. Between September 1982 and October 2006 the Melbourne clinic conducted 1164 CIs with 62 implant failures and explantation (5.3%). Reasons included device failure, (ECSCIFE category C, n = 35, 3.0%), medical complication (ECSCIFE category D, n = 19), characteristics decrement (ECSCIFE category B1, n = 2) and performance decrement (ECSCIFE category B2 n = 6). Forty-nine patients underwent ipsilateral explantation/reimplantation. Auditory performance with the second implant was comparable to the first implant levels. The ECSCIFE provides an easy-to-use classification system for international reporting of CI device failures and explantations, however, detailed information of each implant failure may be lost in the final category, however. Cochlear explantation/reimplantation in Melbourne is a safe and effective procedure to restore patients to their pre-operative best auditory function levels.

Paraganglioma: the role of genetic counselling and radiological screening.

Paragangliomas are rare tumours of the autonomic nervous system that occur in both sporadic and hereditary forms. They are usually benign tumours with low mortality, but can cause significant morbidity related to mass effect. Genetic predisposition to develop paraganglioma can occur within known tumour syndromes and familial tumours tend to present at a younger age and at multiple sites compared to sporadic tumours. Tumours should be diagnosed and excised as early as possible, as studies have shown morbidity to be directly related to tumour size. We present a case of a 14-year-old boy with multiple paraganglioma and a strong family history of paraganglioma. He suffered significant morbidity at resection of an extra-adrenal retroperitoneal tumour due to late diagnosis and was later unable to undergo excision of a head and neck paraganglioma due to its size and relation to neurovascular structures in the neck. We review the current literature on suggested genetic counselling (psychological counselling and DNA analysis) and radiological screening guidelines and recommend that genetic counselling should be offered to all patients with a family history of paraganglioma from the age of 5 years. Those positive for paternal paraganglioma locus gene should then undergo regular radiological screening with MRI.

Hyperventilation in the vestibular clinic: use of the Nijmegen Questionnaire.

The objective of the present study was to determine the prevalence of hyperventilation syndrome in patients seen for vestibular assessment and to assess the clinical utility of the Nijmegen Questionnaire in this group. The Nijmegen Questionnaire and Dizziness Handicap Inventory (DHI) were administered prospectively to a consecutive series of 100 patients identified as candidates for vestibular assessment within the University Hospital Neuro-otology practice. Twenty-three per cent of patients seen for vestibular assessment were diagnosed with hyperventilation syndrome using the Nijmegen Questionnaire. Seventeen of these (74%) would have remained undetected had the Nijmegen questionnaire not been used. No relationship was found between vestibular assessment results and either Nijmegen or DHI scores. A significant correlation was found between DHI scores and Nijmegen Questionnaire scores (rho = 0.348, P = 0.0005). In conclusion, the Nijmegen Questionnaire is a quick, easy to administer and low-impact assessment tool for hyperventilation syndrome and is a useful adjunct to the otological consultation. Diagnosed patients can then be offered breathing control exercises as part of a vestibular rehabilitation programme.

Squamous cell carcinoma of the temporal bone.

Squamous cell carcinoma of the temporal bone is a rare, highly malignant, aggressive disease in an extremely difficult anatomic site with a very poor prognosis. The development of an evidence base for the efficacy of its treatment has been hampered by its rarity. This article reviews current opinion based on the recent literature.

Functional and morphological response of the stria vascularis following a sensorineural hearing loss.

Cochlear endolymph is maintained at a potential of (+)80 mV by an active transport mechanism involving the stria vascularis (SV). This so-called endocochlear potential (EP) is integral to hair cell transduction. We compared the EP with changes in SV area and Na(+),K(+)-ATPase expression following a sensorineural hearing loss. Guinea pigs were deafened using kanamycin and a loop diuretic, and the EP was measured at two, 14, 56, 112 or 224 days following deafening. Auditory brainstem responses were used to confirm that each animal had a severe-profound hearing loss. There was a significant reduction in EP following two days of deafness (normal, 73.5 mV S.E.M.=2.4; deaf, 42.1 mV, S.E.M.=2.8; P<0.0001, t-test). In animals deafened for 14 days the EP had partially recovered (65.2 mV, S.E.M.=5.08), while animals deafened for longer periods exhibited a complete recovery (56 days 80.5 mV, S.E.M.=5.36; 112 days 75.7 mV, S.E.M.=2.71; 224 days 81.0 mV; S.E.M.=6.0). Despite this recovery, there was a systematic reduction in SV area with duration of deafness over the first 112 days of deafness. Significant reductions were localised to the basal turn in animals deafened for two days, but had extended to all turns in animals deafened for 112 days. While there was a significant reduction in strial area, the optical density of Na(+),K(+)-ATPase within the remaining SV was normal. Since the treated animals exhibited essentially a complete elimination of all hair cells, the total K(+) leakage current from the scala media would be expected to be significantly reduced. The large reduction in the extent of the SV after deafening suggests that a reduced strial volume is capable of maintaining a normal EP under conditions of reduced K(+) leakage current.

Expression of Schwann cell-specific proteins and low-molecular-weight neurofilament protein during regeneration of sciatic nerve treated with neurotrophin-4.

Neurotrophin-4 acts as a potent survival factor for subpopulations of motoneurons. To investigate its effect on Schwann cell sheath and axonal proteins during peripheral nerve regeneration, sciatic nerves in adult rats were transected and repaired, and fibrin glue containing neurotrophin-4 injected around the repair site. At 5, 15, 30 and 60 days after repair, 5-mm nerve segments distal to the repair were collected, and western blotting was used to measure myelin-associated glycoprotein, myelin basic protein and low-molecular-weight neurofilament protein. In control groups these dramatically declined at 5 and 15 days then increased from 30 and 60 days. However, in the neurotrophin-4 group there was a significant increase (to several times basal values) in myelin-associated glycoprotein and myelin basic protein at 5-15 days. The relatively small increases (<7%) in Schwann cell numbers suggest that this is mainly due to increased synthesis per cell. The neurotrophin-4 group also showed a small but significant increase at 15 days in low-molecular-weight neurofilament protein, which however remained much lower than basal. We conclude that neurotrophin-4 regulates the expression of myelin-associated glycoprotein, myelin basic protein, and to a lesser extent low-molecular-weight neurofilament protein, during peripheral nerve regeneration.

Neurotrophin-4 delivered by fibrin glue promotes peripheral nerve regeneration.

Neurotrophin-4 (NT-4) is a recently identified neurotrophic factor with potential trophic effects on subpopulations of neurons. Little is known about its role in peripheral nerve regeneration following nerve injury. To investigate this, 48 Sprague-Dawley rats underwent left sciatic nerve transection and immediate repair. Fibrin glue mixed with either NT-4 or vehicle (control) was injected around the nerve repair site. Nerve regeneration was assessed both functionally and histomorphometrically. The results showed that the NT-4-treated group had a significant increase compared with the control in the regeneration distance at 5 days. The sciatic function index was significantly greater in the NT-4 group from 40 to 60 days after nerve repair. Morphometric analysis revealed that nerves treated with NT-4 had significant improvement in the number of regenerated axons, axonal diameter, and myelin thickness. These results suggest that NT-4 is a potent factor improving rat sciatic nerve regeneration.

Parathyroid hormone potentiates nucleotide-induced [Ca2+]i release in rat osteoblasts independently of Gq activation or cyclic monophosphate accumulation. A mechanism for localizing systemic responses in bone.

The regulation of tissue turnover requires the coordinated activity of both local and systemic factors. Nucleotides exist transiently in the extracellular environment, where they serve as ligands to P2 receptors. Here we report that the localized release of these nucleotides can sensitize osteoblasts to the activity of systemic factors. We have investigated the ability of parathyroid hormone (PTH), a principal regulator of bone resorption and formation, to potentiate signals arising from nucleotide stimulation of UMR-106 clonal rat osteoblasts. PTH receptor activation alone did not lead to [Ca(2+)](i) elevation in these cells, indicating no G(q) coupling, however, activation of G(q)-coupled P2Y(1) receptors resulted in characteristic [Ca(2+)](i) release. PTH potentiated this nucleotide-induced Ca(2+) release, independently of Ca(2+) influx. PTH-(1-31), which activates only G(s), mimicked the actions of PTH-(1-34), whereas PTH-(3-34), which only activates G(q), was unable to potentiate nucleotide-induced [Ca(2+)](i) release. Despite this coupling of the PTHR to G(s), cAMP accumulation or protein kinase A activation did not contribute to the potentiation. 3-Isobutyl-1-methylxanthine, but not forskolin effectively potentiated nucleotide-induced [Ca(2+)](i) release, however, further experiments proved that cyclic monophosphates were not involved in the potentiation mechanism. Costimulation of UMR-106 cells with P2Y(1) agonists and PTH led to increased levels of cAMP response element-binding protein phosphorylation and a synergistic effect was observed on endogenous c-fos gene expression following costimulation. In fact the calcium responsive Ca/cAMP response element of the c-fos promoter alone was effective at driving this synergistic gene expression. These findings demonstrate that nucleotides can provide a targeted response to systemic factors, such as PTH, and have important implications for PTH-induced signaling in bone.

Pediatric hospitalists in Canada and the United States: a survey of pediatric academic department chairs.

OBJECTIVES: To document the prevalence and practice patterns of pediatric hospitalists in academic centers in Canada and the United States; to characterize academic pediatric department chairs' definition of the term hospitalist; and to characterize pediatric department chairs' views of the training requirements for pediatric hospitalists. METHODS: A 14-item questionnaire was sent to all 145 pediatric department chairs from Canada and the United States during the fall of 1998. We defined hospitalists as physicians spending at least 25% of their time in inpatient care. RESULTS: Of the 145 eligible pediatric chairs, 128 (89%) responded (United States, 111/126; Canada, 14/16; Puerto Rico, 3/3). Ninety-nine (77%) of 128 pediatric chairs either have (64/128) or are planning to have (35/128) hospitalists in their institutions. Within academic programs with hospitalists, 82% of hospitalists currently work on general pediatric wards. Two thirds of hospitalists teach, 50% provide outpatient care, 50% have administrative duties, and 44% conduct research. One hundred eight (84%) of 128 believe that hospitalists should spend at least 50% of their time in inpatient care. Less than one third (30%) of pediatric chairs believe that hospitalists require training not currently provided in residency. CONCLUSIONS: A large proportion of academic pediatric centers either employed or planned to employ hospitalists in 1998. Pediatric academic department chairs do not see a need for training beyond residency for hospitalists. Further studies should address how pediatric hospitalists affect quality of care, cost, and patient satisfaction.

Can falls on stairs result in small intestine perforations?

OBJECTIVE: Abusive parents often report that a fall on stairs resulted in their children's injuries. This review explores whether there is any evidence in the medical literature that a fall on stairs could be a plausible explanation for a small intestine perforation. METHODOLOGY: The English-language medical literature was searched by Medline, for a 29-year period (1970-1998), for reports of the types of injuries sustained in falls on stairs and for reports of the types of blunt abdominal trauma that result in small intestine perforations. Articles that exclusively focused on infant walker injuries or the elderly were excluded. Duodenal, jejunal, and ileal perforations were included, whereas intestinal hematomas and undescribed intestinal injuries were excluded. All types of injuries to the stomach, colon, and rectum were excluded. RESULTS: Falls on stairs were not reported to be a cause for any of the 312 cases of small intestine perforations reviewed. There were no reports of any intraabdominal injuries, including small intestine perforations, in any of the 677 cases of falls on stairs reviewed. Falls on stairs rarely resulted in any type of truncal injury. CONCLUSIONS: Although falls on stairs have been reported to be the most common cause of injury in childhood, no evidence was found to support the contention that an unobstructed fall on stairs could be consistent with perforation of the small intestine.

The phylogeny of the Asteridae sensu lato based on chloroplast ndhF gene sequences.

A phylogenetic study of Asteridae sensu lato was conducted based on chloroplast ndhF gene sequences for 116 ingroup and 13 outgroup species. Prior molecular studies based on rbcL sequences identified terminal groups corresponding to families, but were unable to resolve relationships among them. These results are largely consistent with earlier rbcL studies, but provide much greater resolution and stronger bootstrap support throughout the tree. The parsimony analysis found eight equally parsimonious trees, all of which recognize four major clades with the following relationship: (Cornales (Ericales (Euasterids I, Euasterids II))). Euasterids I includes (Garryales ((Solanales, Boraginaceae) (Gentianales, Lamiales))), although with weak support for relationships among the named clades. Euasterids II includes (Aquifoliales (Asterales (Apiales, Dipsacales))) with strong support for these relationships. Relationships within Ericales are weakly supported and merit further attention.

A CD36-binding peptide from thrombospondin-1 can stimulate resorption by osteoclasts in vitro.

Thrombospondin-1 (TSP-1), purified from platelets, stimulates resorption by avian osteoclasts in an in vitro resorption assay. TSP-1 binds to a number of different cellular receptors via different domains of the molecule and several short receptor-binding sequences have been identified within the TSP-1 molecule. In this study, we have used synthetic peptides representing these various sequences in order to identify the cellular receptor and TSP domain responsible for stimulation of resorption. We show that one peptide CSVTCG, which represents the CD36-binding region of TSP-1, stimulates resorption in a fashion similar to the intact molecule, while the peptides RGDS, RFYVVMWK, and RFYVVM, representing other cell-binding domains of TSP, have no effect on resorption. Using RT-PCR and immunoblotting, we further demonstrate expression of CD36 in human osteoclastoma (giant cell tumour), primary human bone derived cells, and clonal osteoblastic cells. These studies suggest that CD36 is involved in regulation of resorption by osteoclasts and is the receptor responsible for the resorption-promoting effects of TSP-1.

Extracellular ATP activates multiple signalling pathways and potentiates growth factor-induced c-fos gene expression in MCF-7 breast cancer cells.

In the human breast cancer cell line MCF-7, the nucleotides ATP gamma S and UTP, acting extracellularly through the purinergic receptor P2Y(2), lead to elevated intracellular calcium levels and increased proliferation. ATP gamma S and UTP treatment of MCF-7 cells activated transcription of the immediate early gene c-fos, an important component in the response to proliferative stimulation. c-fos induction was enhanced by co-treatment with ATP gamma S and a variety of proliferative agents including growth factors, tumour promoters and stress. Stimulation with ATP gamma S or epidermal growth factor (EGF) led to extracellular signal-regulated kinase (ERK) activation and phosphorylation of the transcription factors CREB and Elk-1. Co-stimulation synergistically activated fos expression and notably led to increased levels of ERK, CREB and EGF receptor phosphorylation, as well as hyperphosphorylation of ternary complex factor. Nevertheless, the ERK pathway does not fully account for this synergy, since fos induction was differentially sensitive to the MEK inhibitor U0126, indicating that these two agonists signal differently to this immediate early gene. Thus, extracellular nucleotides co-operate with growth factors to activate genes linked to the proliferative response in MCF-7 cells through activation of specific purinergic receptors, which thereby represent important potential targets for arresting the neoplastic progression of breast cancer cells.

Expression of members of the thrombospondin family by human skeletal tissues and cultured cells.

We have previously shown that the multifunctional platelet glycoprotein thrombospondin-1 (TSP-1) promotes resorption in an in vitro resorption assay. However, TSP-1 is one of a family of multifunctional TSP molecules, and the current study was undertaken to investigate whether it is TSP-1 or another TSP family member which may be involved in regulation of resorption in vivo. RT-PCR was performed on cultured human bone cells, cultured human chondrocytes, and three separate samples of human osteoclastoma tissue using primers specific for each TSP family member. mRNA for TSP-2 was detected in almost all samples, and significantly in all osteoclastomas in the above tissues, while TSP-1 was detected less frequently and was only seen in one of three osteoclastomas. TSP-3, -4, and COMP were detected only in a minority of cases. These results indicate that TSP-2 is the most common TSP family member found in skeletal tissues and that TSP-2, rather than TSP-1, may be the molecule responsible for promoting resorption in vivo.

Classification, origins, and patterns of diversification in New ZealandCarmichaelinae (Fabaceae).

Analysis of ITS sequences provides support for a clade that includes Carmichaelia, Clianthus, Montigena, and Swainsona. We provide a node-based definition and recommend that this clade be called Carmichaelinae. Results suggest that Carmichaelinae are derived from northern hemisphere Astragalinae. The clade has extensively radiated in Australia, and two independent lineages have diversified in New Zealand. The New Zealand lineages differ in species richness. One lineage consists of 24 species placed in Carmichaelia and Clianthus, while the other corresponds to the monotypic genus Montigena. The pattern of relationships inferred from ITS sequences suggests that the New Zealand radiation was recent and possibly accompanied episodes of mountain-building and glaciation.