Genetic variation in blue whales in the eastern pacific: implication for taxonomy and use of common wintering grounds.

Many aspects of blue whale biology are poorly understood. Some of the gaps in our knowledge, such as those regarding their basic taxonomy and seasonal movements, directly affect our ability to monitor and manage blue whale populations. As a step towards filling in some of these gaps, microsatellite and mtDNA sequence analyses were conducted on blue whale samples from the Southern Hemisphere, the eastern tropical Pacific (ETP) and the northeast Pacific. The results indicate that the ETP is differentially used by blue whales from the northern and southern eastern Pacific, with the former showing stronger affinity to the region off Central America known as the Costa Rican Dome, and the latter favouring the waters of Peru and Ecuador. Although the pattern of genetic variation throughout the Southern Hemisphere is compatible with the recently proposed subspecies status of Chilean blue whales, some discrepancies remain between catch lengths and lengths from aerial photography, and not all blue whales in Chilean waters can be assumed to be of this type. Also, the range of the proposed Chilean subspecies, which extends to the Galapagos region of the ETP, at least seasonally, perhaps should include the Costa Rican Dome and the eastern North Pacific as well.

[Pulmonary lymphangioleiomyomatosis: From pathogenesis to management]. / Lymphangioléiomyomatose pulmonaire : de la physiopathologie à la prise en charge.

INTRODUCTION: Pulmonary lymphangioleiomyomatosis (LAM) is a rare disease affecting mainly young women. BACKGROUND: The respiratory manifestations are characterized by a progressive cystic destruction of the lung parenchyma. Extrapulmonary involvement includes benign renal tumours called angiomyolipomas and abdominal lymphatic masses called lymphangioleiomyomas. At the pathological level, the cellular proliferation found in LAM is in part due to the presence of mutations in the tumour suppressor genes TSC1 and TSC2 (Tuberous Sclerosis Complex). These mutations lead to the activation of the mTOR pathway, which is currently the main therapeutic target. mTOR inhibitors such as sirolimus or everolimus have shown a beneficial effect on the decline in pulmonary function and a reduction of angiomyolipoma size, but are necessary in only some patients. PERSPECTIVES: LAM cells have migratory properties mediated by the formation of new lymphatic vessels. They are also able to secrete metalloproteases, which enhance their invasiveness. Moreover, the expression of estrogen and progesterone receptors by LAM cells suggests a possible role for sex hormones in the pathogenesis of the disease. CONCLUSION: A better understanding of mTOR-independent mechanisms would allow the development of novel therapeutic approaches.

Advances in the study of transmissible respiratory tumours in small ruminants.

Sheep and goats are widely infected by oncogenic retroviruses, namely Jaagsiekte Sheep RetroVirus (JSRV) and Enzootic Nasal Tumour Virus (ENTV). Under field conditions, these viruses induce transformation of differentiated epithelial cells in the lungs for Jaagsiekte Sheep RetroVirus or the nasal cavities for Enzootic Nasal Tumour Virus. As in other vertebrates, a family of endogenous retroviruses named endogenous Jaagsiekte Sheep RetroVirus (enJSRV) and closely related to exogenous Jaagsiekte Sheep RetroVirus is present in domestic and wild small ruminants. Interestingly, Jaagsiekte Sheep RetroVirus and Enzootic Nasal Tumour Virus are able to promote cell transformation, leading to cancer through their envelope glycoproteins. In vitro, it has been demonstrated that the envelope is able to deregulate some of the important signaling pathways that control cell proliferation. The role of the retroviral envelope in cell transformation has attracted considerable attention in the past years, but it appears to be highly dependent of the nature and origin of the cells used. Aside from its health impact in animals, it has been reported for many years that the Jaagsiekte Sheep RetroVirus-induced lung cancer is analogous to a rare, peculiar form of lung adenocarcinoma in humans, namely lepidic pulmonary adenocarcinoma. The implication of a retrovirus related to Jaagsiekte Sheep RetroVirus is still controversial and under investigation, but the identification of an infectious agent associated with the development of lepidic pulmonary adenocarcinomas might help us to understand cancer development. This review explores the mechanisms of induction of respiratory cancers in small ruminants and the possible link between retrovirus and lepidic pulmonary adenocarcinomas in humans.

Can DVD simulations provide an effective alternative for paramedic clinical placement education?

OBJECTIVE: This study aimed to evaluate the usability of DVD simulations, the impact on student learning satisfaction and the potential for using DVD simulations to reduce the clinical placement burden on the current healthcare system. The clinical DVD simulations were underpinned by interprofessional educational principles that supported clinical placements for paramedic students. METHOD: Eleven DVD simulations were developed by academic staff members from Monash University with input and feedback from a team of healthcare professionals. Students (N = 97) from the Bachelor of Emergency Health at Monash University viewed the DVD simulations. Students' perceptions, attitudes and thoughts about the clinical relevance of the simulations were assessed by completing a standardised self-report 7-point Likert scale questionnaire (7 indicating the highest satisfaction score). Qualitative data assessing if and how the DVD simulations had influenced paramedic students' clinical placement learning experiences were also collected via two focus groups (n = 6). RESULTS: Overall, paramedic students positively perceived the DVD simulations with relation to learning satisfaction (mean (SD) 5.14 (1.14), 95% CI 4.91 to 5.37) and information processing quality (mean (SD) 5.50 (0.83), 95% CI 5.33 to 5.67). The simulations maintained students' attention and concentration (mean (SD) 4.35 (0.95), 95% CI 4.15 to 4.54) and provided clinical authenticity and relevance to practice (mean (SD) 4.27 (0.65), 95% CI 4.14 to 4.40). A number of themes emerged from the focus group data including the impact on employment, greater appreciation of healthcare teamwork and notion of interdisciplinary teamwork, the fact that DVD simulations have the capacity to replace some clinical placement rotations and should be integrated into standard curriculum, and that varying amounts of learning wastage occur during clinical placements. CONCLUSIONS: DVD simulations with an interprofessional education focus were developed. Paramedic students reported the simulations as being educationally, professionally and clinically relevant. The students also identified some aspects of current clinical placements that may be replaced by using DVD simulations. The cost benefit of using interprofessional DVD simulations to supplement and replace certain clinical placement rotations should be investigated further.

Cardiac troponin I is a sensitive, specific biomarker of cardiac injury in laboratory animals.

This study directly demonstrates that cardiac troponin I (cTnI) is a sensitive, specific, and persistent biomarker in laboratory animals. Histopathological and pathophysiological cardiac changes in dogs, rats and mice correlated with increased serum cTnI with various cardiac inotropic agents, and cardiotoxic drugs and with cardiac arrhythmias, tachycardia, cardiac effusion with dyspnoea, and ageing. A comparison of six immunoassays for cTnI and cardiac troponin T (cTnT) to detect and monitor cardiac injury in a rodent model indicated that enzyme-linked immunosorbent (Life Diagnostics Inc and TriChem Resources Inc, West Chester, Philadelphia, USA) and Immulite (Diagnostic Products Corporation, Llanberis, UK) assays had low sensitivity and less than 1% of the dynamic range of Centaur (Bayer Healthcare Diagnostics, Newbury, UK) cTnI and Elecsys (Roche Diagnostics, Basel, Switzerland) and M8 (Bioveris Europe, Whitney, UK) cTnT assays. In dogs, however, the Immulite assay was effective and correlated with the Centaur. Serum concentrations were highly correlated but 10-fold lower for cTnT compared with cTnI with cardiac injury. Centaur assay also detected cTnI in myocardium from marmosets, swine, cattle, and guinea pigs, indicating it to be candidate cardiac biomarker for these species as well. Purified rat cTnI was 50% more reactive than purified human cTnI in the Centaur assay. In the rat, an age- and gender-dependent variation in serum cTnI was found. Male rats aged six and eight months had a 10-fold greater serum cTnI than age-matched females and three-month-old rats. These increases correlated with minimal histopathological change. Isoproterenol-induced serum cTnI increased up to 760-fold the minimal detectable concentration of 0.07 microg/L, within 4-6 h and decreased with a half-life of 6 h, with an expected return to baseline of 60 h. Severity of histopathological change correlated with serum cTnI during the ongoing injury.

Telomerase activation in a model of lung adenocarcinoma.

Ovine pulmonary adenocarcinoma (OPA) is a lung cancer strikingly similar to the pneumonic-type mixed invasive adenocarcinoma with a predominant bronchioloalveolar component in humans. Telomerase activity in OPA and the potential involvement of the kinase Akt in telomerase activation and regulation of cell proliferation were investigated. Lung tissues were collected from sheep with a histopathological diagnosis of OPA or controls. Epithelial cell cultures were derived in vitro from lung tissues. Telomerase activity was evaluated using the telomeric repeat amplification protocol method. Phosphorylation of Akt was detected by Western blotting. Telomerase activity was significantly higher in OPA lung tissues compared to control lung tissues. A high telomerase activity was detected in eight out of 12 (67%) primary cell cultures derived from tumours. A high level of expression of phosphorylated Akt was found in 10 out of 27 (37%) tumours, with abolition of Akt activation in response to epidermal growth factor stimulation demonstrated in primary cell cultures derived from tumours. Telomerase activation takes place in ovine pulmonary adenocarcinoma tumour cells and may be partly attributable to Akt activation. Telomerase may inhibit cellular senescence and contribute to the accumulation of tumour cells in mixed adenocarcinoma with a bronchioloalveolar component. Further work is necessary to identify alternative signalling pathways of telomerase activation in tumours.

[JSRV (jaagsiekte sheep retrovirus) and ovine pulmonary carcinoma]. / JSRV (jaagsiekte sheep retrovirus) et cancer du poumon associé chez le mouton.

JSRV (jaagsiekte sheep retrovirus) is a betaretrovirus, infecting small ruminants. This virus is responsible for the development of pulmonary adenocarcinoma, by the transformation of epithelial cells of the bronchioli and alveoli. This animal cancer is related to human bronchioloalveolar cancer (BAC), a specific form of human lung cancer for which a viral etiology has been proposed for several decades. In small ruminants JSRV interacts with the cells through the Hyal2 receptor. JSRV genome is simple and does not contain already known oncogene. It is now well established that the envelope protein is oncogenic by itself, via the cytoplasmic domain of the transmembrane glycoprotein and some domains of the surface glycoprotein. Activation of the PI3K/Akt and MAPK pathways participates to the envelope-induced transformation. The tumour development is associated with telomerase activation.

Ex vivo propagation of infectious sheep scrapie agent in heterologous epithelial cells expressing ovine prion protein.

Transmissible spongiform encephalopathies, or prion diseases, are fatal degenerative disorders of the central nervous system that affect humans and animals. Prions are nonconventional infectious agents whose replication depends on the host prion protein (PrP). Transmission of prions to cultured cells has proved to be a particularly difficult task, and with a few exceptions, their experimental propagation relies on inoculation to laboratory animals. Here, we report on the development of a permanent cell line supporting propagation of natural sheep scrapie. This model was obtained by stable expression of a tetracycline-regulatable ovine PrP gene in a rabbit epithelial cell line. After exposure to scrapie agent, cultures were repeatedly found to accumulate high levels of abnormal PrP (PrPres). Cell extracts induced a scrapie-like disease in transgenic mice overexpressing ovine PrP. These cultures remained healthy and stably infected upon subpassaging. Such data show that (i) cultivated cells from a nonneuronal origin can efficiently replicate prions; and (ii) species barrier can be crossed ex vivo through the expression of a relevant PrP gene. This approach led to the ex vivo propagation of a natural transmissible spongiform encephalopathy agent (i.e., without previous experimental adaptation to rodents) and might be applied to human or bovine prions.

Evidence that Wallerian degeneration and localized axon degeneration induced by local neurotrophin deprivation do not involve caspases.

The selective degeneration of an axon, without the death of the parent neuron, can occur in response to injury, in a variety of metabolic, toxic, and inflammatory disorders, and during normal development. Recent evidence suggests that some forms of axon degeneration involve an active and regulated program of self-destruction rather than a passive "wasting away" and in this respect and others resemble apoptosis. Here we investigate whether selective axon degeneration depends on some of the molecular machinery that mediates apoptosis, namely, the caspase family of cysteine proteases. We focus on two models of selective axon degeneration: Wallerian degeneration of transected axons and localized axon degeneration induced by local deprivation of neurotrophin. We show that caspase-3 is not activated in the axon during either form of degeneration, although it is activated in the dying cell body of the same neurons. Moreover, caspase inhibitors do not inhibit or retard either form of axon degeneration, although they inhibit apoptosis of the same neurons. Finally, we cannot detect cleaved substrates of caspase-3 and its close relatives immunocytochemically or caspase activity biochemically in axons undergoing Wallerian degeneration. Our results suggest that a neuron contains at least two molecularly distinct self-destruction programs, one for caspase-dependent apoptosis and another for selective axon degeneration.

Corneal stromal sequestration and keratoconjunctivitis sicca in a horse.

A 19-year-old Shetland pony presented with unilateral ocular discomfort and abnormal ocular appearance. Keratoconjunctivitis sicca, ulcerative keratitis and brown discoloration of the corneal stroma were identified on ophthalmic examination. The etiology of keratoconjunctivitis sicca was not determined in this case. For practical and financial reasons, the owners requested enucleation of the affected eye. Histopathologic examination revealed extensive loss of corneal epithelium overlying a zone of hypereosinophilic, degenerate, and necrotic corneal stroma. This well-circumscribed region of corneal stromal sequestration was surrounded by stromal vascularization, and an intense inflammatory, predominantly polymorphonuclear, cellular infiltrate. The clinical and histopathologic features of this case were considered remarkably similar to those observed in feline corneal stromal sequestration.

CAMs and FGF cause a local submembrane calcium signal promoting axon outgrowth without a rise in bulk calcium concentration.

Binding of basic fibroblast growth factor (bFGF) and cell adhesion molecules to the nerve cell membrane promotes axon outgrowth. This response can be blocked by antagonists of voltage-gated calcium channels, yet no change of cytosolic calcium concentration in the growth cone can be detected upon binding of the growth factor bFGF or the cell adhesion molecule L1. Using barium as a charge carrier, we show that bFGF and L1 open a calcium influx pathway in growth cones of rat sensory neurons without changing the membrane voltage. L1 does not activate influx in cells expressing a dominant negative mutant of the fibroblast growth factor receptor (FGFR) tyrosine kinase. FGFR-activated influx is blocked by specific antagonists of L- and N-type voltage-gated calcium channels and by an inhibitor of diacylglycerol lipase. We propose that both L1 and bFGF act via the FGFR to generate polyunsaturated fatty acids which in turn cause calcium channels to flicker open and shut. Short-lived domains of raised calcium at the cytosolic mouth of open channels activate axon outgrowth without raising bulk cytosolic calcium concentration. In confirmation of this model, the rapidly-acting calcium buffer BAPTA is significantly more effective at blocking FGF-induced axon outgrowth when compared with the slower buffer EGTA. Generation of short-lived calcium domains may provide a crucial mechanism for axon guidance during development and for promoting regeneration of damaged axons.

A comparison of serum ionized calcium concentrations and serum total calcium concentrations in dogs with lymphoma.

Serum ionized calcium and total calcium concentrations were measured in 16 dogs with lymphoma and 49 healthy control dogs. Blood samples for all determinations of ionized calcium were collected into tubes containing silicone separator gel and processed under a closed anaerobic system. An ionized calcium analyser with ion selective electrodes was used to determine pH, ionized calcium and ionized calcium adjusted to pH 7.4. Reference ranges for ionized calcium (iCa pH 7.4) of 1.30-1.46 mmol/l, and for total calcium of 2.37-2.82 mmol/l were established. A stronger correlation (r = 0.85) was found between measured ionized calcium and total calcium levels in dogs with lymphoma and hypercalcaemia than in those with lymphoma and normocalcaemia (r = 0.64). Measurement of serum ionized calcium was diagnostically concordant with the measurement of serum total calcium in the determination of calcium status in all dogs with lymphoma. Serum ionized calcium did not provide a diagnostic advantage over total calcium in the detection of hypercalcaemia of malignancy in these dogs.

Evaluation of three pancreas specific protein assays, TLI(trypsin-like immunoreactivity), PASP (pancreas specific protein) and CA 19-9 (glycoprotein) for use in the diagnosis of canine pancreatitis.

Three radioimmunoassays (RIA) for the pancreas specific proteins TLI, PASP and CA 19-9 were evaluated in serum from normal control dogs (n = 40) and dogs with pancreatitis (n = 20). Statistically significant differences (P < 0.05) were found for serum TLI and PASP levels between the control and pancreatitis groups. However, only 3/20 dogs with pancreatitis had serum TLI concentrations greater than the highest concentration in control dogs. Concentrations of PASP in serum were higher in 15/20 dogs with pancreatitis than in the control dogs. The magnitude of the increase in concentrations of PASP in pancreatitis was small in the majority of cases. Thus these assays are of limited clinical value in the diagnosis of pancreatitis. There was no cross-reactivity with dog serum in the CA 19-9 assay.

Differences in electrophoresis patterns between plasma albumins of the cockatiel (Nymphicus hollandicus) and the chicken (Gallus gallus domesticus).

Unique migration patterns for chicken and cockatiel albumins were detected when electrophoresis was performed using cellulose acetate and agarose gel support media. Cockatiel albumin migrated to a position equivalent to chicken alpha globulins, while the migration of cockatiel prealbumin was similar to that of chicken albumin. A chicken prealbumin band was not detected. Cockatiel and chicken albumins purified by affinity chromatography had similar migration patterns when electrophoresis was performed under denaturing conditions [sodium dodecyl sulphate (SDS) ] in 7% polyacrylamide gel (PAGE). The molecular weights of both albumins were similar, and were estimated to be approximately 66,000 Da when compared to known molecular weight markers. The different migration patterns were attributed to variations in conformation and surface charge distribution of albumin molecules between the two species. The experimental and clinical consequences of these findings are briefly discussed.

Alkaline phosphatase bone isoenzyme and osteocalcin in the serum of hyperthyroid cats.

The effect of hyperthyroidism on serum markers for increased bone metabolism and turnover was evaluated in 36 cats with elevated serum levels of thyroxine and alkaline phosphatase. Serum was analyzed for total and ionized calcium and phosphorous. Alkaline phosphatase isoenzymes were separated by agarose gel electrophoresis and osteocalcin was measured by radioimmunoassay. Values for hyperthyroid cats were compared with those for healthy cats. Alkaline phosphatase bone isoenzyme was markedly increased in all 36 hyperthyroid cats. Osteocalcin was increased in 44% of the cats. There was no correlation among the magnitude of increase in alkaline phosphatase bone isoenzyme, osteocalcin, and serum thyroxine concentrations. Increased serum phosphorus was found in 35% of the cats. Total calcium was within the reference range in all cats, while 50% of the cats had reduced levels of serum ionized calcium. We conclude that hyperthyroid cats do have altered bone metabolism, although it is usually clinically insignificant.

Comparison of media and sera used for in vitro culture of equine superficial digital flexor tendon explants.

OBJECTIVE: To compare the effects of different commercial nutrient media and sera on protein synthesis and maintenance of cellular density in cultures of the equine superficial digital flexor tendon (SDFT). ANIMALS: 8 healthy 2- to 4-year-old horses. PROCEDURE: First Dulbecco's modified Eagle's medium, Ham's F12 nutrient mixture, RPMI 1640 medium, minimum essential medium with Earle's salts, minimum essential medium with Hanks' salts, and a Dulbecco's modified Eagle's medium/Ham's F12 nutrient mixture with 10% fetal bovine serum (FBS) were compared. Then FBS, fetal equine serum, and donor horse serum, each at 5, 10, and 15% in RPMI 1640 medium, were compared. Explants were cultured in roller bottles at 37 C and aerated (50% O2/45% N2/5% CO2) daily. Rates of [3H]-proline incorporation were used as a measure of the rates of total protein and collagen synthesis on days 13 and 28. Matrix cellular density of explants at days 14 and 28 was measured by computerized image analysis. RESULTS: Equine SDFT explants were cultured in all media for up to 4 weeks. Proline incorporation was greatest in Ham's F12 nutrient mixture and in RPMI 1640 medium, with the concentration of proline in medium correlating to the in vitro response. Total proline incorporation was greater in 15% FBS than in 5 or 10% FBS. Other differences among sera were not detected. Matrix cell density in 15% donor horse serum was equivalent to that in uncultured controls and higher than that in most other sera at week 2. CONCLUSION: The in vitro SDFT culture system described may be used in future studies to enhance knowledge of the biological and biochemical characteristics of intrinsic tendon healing.

Conformational study in water by NMR and molecular modeling of cyclic glutamic acid analogues as probes of vitamin K dependent carboxylase.

The conformational analysis of four glutamic acid analogues containing a cyclopentyl or cyclohexyl ring, substituted in position 1 by a BOC protected amino group and a methyl ester group and in position 3 by a free carboxylate group, has been carried out in an aqueous environment, by 1H and 13C NMR spectroscopy, and molecular dynamics (MD). Their structural properties were under investigation for a structure-activity relationship analysis to determine the preferred conformation in the carboxylase active site. For each compounds, resulting conformations from NMR and MD data were analyzed and classified according to the dihedral angles chi 1 and chi 2, the distances and the spatial distribution involving charged or substituted C- and N-terminal groups. A reduced number of conformational families were found to be in qualitative agreement with NMR and MD data. A comparison between these different classes of the active and nonactive derivatives was achieved.

Ascites and hepatic cirrhosis in a cockatiel (Nymphicus hollandicus).

A mature, female cockatiel (Nymphicus hollandicus) was examined because of respiratory difficulties. Clinical and laboratory findings included ascites and evidence of hepatic disease (i.e., increased plasma bile acid concentrations, aspartate aminotransferase, and alkaline phosphatase activities). Plasma protein electrophoresis results were consistent with chronic-active inflammation. The albumin-to-globulin (A:G) ratio, calculated from plasma electrophoresis, was 0.3. Postmortem examination revealed severe hepatic fibrosis and a diffuse, interstitial, granulomatous lipid pneumonia.