Multi-omics for studying and understanding polar life.

Polar ecosystems are experiencing amongst the most rapid rates of regional warming on Earth. Here, we discuss 'omics' approaches to investigate polar biodiversity, including the current state of the art, future perspectives and recommendations. We propose a community road map to generate and more fully exploit multi-omics data from polar organisms. These data are needed for the comprehensive evaluation of polar biodiversity and to reveal how life evolved and adapted to permanently cold environments with extreme seasonality. We argue that concerted action is required to mitigate the impact of warming on polar ecosystems via conservation efforts, to sustainably manage these unique habitats and their ecosystem services, and for the sustainable bioprospecting of novel genes and compounds for societal gain.

Chromobacteriosis (Chromobacterium violaceum) in a calf from Brazil - case report / Cromobacteriose (Chromobacterium violaceum) em um bezerro do Brasil - relato de caso

Chromobacterium violaceum is a rare opportunistic pathogen that causes highly fatal infections in domestic animals and humans. This report describes a fatal case suggestive of septicemia in a four-day-old female calf with chromobacteriosis. The calf had suppurative omphalophlebitis, suppurative fibrinous polyarthritis, anterior uveitis with bilateral fibrin deposition, fibrinous peritonitis, lymph node abscess and multifocal lymphocytic and neutrophilic encephalitis with multifocal hemorrhages. C. violaceum was isolated from the spleen and peri-renal lymph node and its identity was confirmed by PCR and sequencing. The pathogen was sensitive to azithromycin, gentamicin, enrofloxacin, norfloxacin, marbofloxacin, ciprofloxacin, erythromycin, sulphazotrim, fluorfenicol, tetracycline and doxycycline as well as resistant to penicillin, ampicillin, vancomycin, amoxicillin, amoxicillin + clavulanic acid, cephalothin, cephalexin, oxacillin, B polymyxin, neomycin and bacitracin. This is the first report of chromobacteriosis in a calf from Brazil.(AU)

Chromobacterium violaceum é um patógeno oportunista raro, que causa infecção fatal em animais domésticos e em humanos. Este relato descreve um caso fatal suspeito de septicemia em um bezerro de quatro dias, fêmea, infectado por C. violaceum. O bezerro apresentava onfaloflebite supurativa, poliartrite supurativa fibrinosa, uveíte anterior com deposição bilateral de fibrina, peritonite fibrinosa, abscesso de linfonodos e encefalite multifocal linfocítica e neutrofílica com áreas hemorrágicas multifocais. C. violaceum foi isolado no baço e no linfonodo, e sua identidade foi confirmada por PCR e sequenciamento. O patógeno foi sensível aos antibióticos azitromicina, gentamicina, enrofloxacina, norfloxacina, marbofloxacina, ciprofloxacina, eritromicina, sulfazotrim, florfenicol, tetraciclina, doxiciclina e foi resistente à penicilina, ampicilina, vancomicina, amoxicilina, amoxicilina + ácido clavulânico, cefalotina, cefalexina, oxacilina, polimixina B, neomicina e bacitracina. Este é o primeiro relato de cromobacteriose em bezerro no Brasil.(AU)

Congenital Segmental Absence of Tracheal Rings in a Kitten.

Congenital anomalies of the trachea are rare in cats. This article reports segmental absence of tracheal cartilage rings in a kitten. An 8-month-old female kitten was presented with a history of weight loss and respiratory distress for 2 months. Radiographs of the thorax demonstrated a large air-filled sac suggestive of pneumomediastinum. No cartilaginous structures were evident radiographically over the caudal portion of the trachea. At necropsy examination, approximately 2 cm from the carina, a 3 cm segment of the trachea lacked cartilaginous rings. The clinical and morphological features of this lesion were similar to those described in human and canine cases of congenital segmental absence of tracheal rings.

Clinicopathological Phenotype of Autosomal Recessive Cholesterol Deficiency in Holstein Cattle.

BACKGROUND: Cholesterol deficiency (CD), a newly identified autosomal recessive genetic defect in Holstein cattle, is associated with clinical signs of diarrhea, failure to thrive, and hypocholesterolemia. HYPOTHESIS/OBJECTIVES: The objective is to describe the clinicopathological phenotype of affected Holstein cattle homozygous for the causative apolipoprotein B gene (APOB) mutation. ANIMALS: Six Holstein cattle, 5 calves with a clinical history of chronic diarrhea, and 1 heifer with erosions in the buccal cavity and neurologic symptoms were admitted to the Clinic for Ruminants. METHODS: This case review included a full clinical examination, a complete blood count, blood chemistry, and measurements of cholesterol and triglycerides. The animals were euthanized and necropsied. A PCR-based direct gene test was applied to determine the APOB genotype. RESULTS: All 6 animals were inbred, could be traced back to the sire Maughlin Storm, and were confirmed homozygous for the APOB mutation. The clinical phenotype included poor development, underweight, and intermittent diarrhea in the calves, and neurologic signs in the heifer included hypermetria and pacing. Hypocholesterolemia and low triglycerides concentrations were present in all animals. The pathological phenotype of all animals was steatorrhea with enterocytes of the small intestine containing intracytoplasmic lipid vacuoles. The peripheral nervous system of the heifer displayed degenerative changes. CONCLUSIONS AND CLINICAL IMPORTANCE: Suspicion of CD in Holstein cattle is based on the presence of chronic diarrhea with no evidence of primary infections. Confirmation of the associated APOB gene mutation is needed. Additionally, the heifer demonstrated primarily signs of neurologic disease providing an unexpected phenotype of CD.

A transposable element insertion in APOB causes cholesterol deficiency in Holstein cattle.

Cholesterol deficiency, a new autosomal recessive inherited genetic defect in Holstein cattle, has been recently reported to have an influence on the rearing success of calves. The affected animals show unresponsive diarrhea accompanied by hypocholesterolemia and usually die within the first weeks or months of life. Here, we show that whole genome sequencing combined with the knowledge about the pedigree and inbreeding status of a livestock population facilitates the identification of the causative mutation. We resequenced the entire genomes of an affected calf and a healthy partially inbred male carrying one copy of the critical 2.24-Mb chromosome 11 segment in its ancestral state and one copy of the same segment with the cholesterol deficiency mutation. We detected a single structural variant, homozygous in the affected case and heterozygous in the non-affected carrier male. The genetic makeup of this key animal provides extremely strong support for the causality of this mutation. The mutation represents a 1.3kb insertion of a transposable LTR element (ERV2-1) in the coding sequence of the APOB gene, which leads to truncated transcripts and aberrant splicing. This finding was further supported by RNA sequencing of the liver transcriptome of an affected calf. The encoded apolipoprotein B is an essential apolipoprotein on chylomicrons and low-density lipoproteins, and therefore, the mutation represents a loss of function mutation similar to autosomal recessive inherited familial hypobetalipoproteinemia-1 (FHBL1) in humans. Our findings provide a direct gene test to improve selection against this deleterious mutation in Holstein cattle.

Heterogeneous accretion and the moderately volatile element budget of Earth.

Several models exist to describe the growth and evolution of Earth; however, variables such as the type of precursor materials, extent of mixing, and material loss during accretion are poorly constrained. High-precision palladium-silver isotope data show that Earth's mantle is similar in 107Ag/109Ag to primitive, volatile-rich chondrites, suggesting that Earth accreted a considerable amount of material with high contents of moderately volatile elements. Contradictory evidence from terrestrial chromium and strontium isotope data are reconciled by heterogeneous accretion, which includes a transition from dominantly volatile-depleted to volatile-rich materials with possibly high water contents. The Moon-forming giant impact probably involved the collision with a Mars-like protoplanet that had an oxidized mantle, enriched in moderately volatile elements.

Clinical validation of the Helikit: a 13C urea breath test used for the diagnosis of Helicobacter pylori infection.

OBJECTIVE: To assess the clinical performance of the Helikit, a 13C urea breath test, in the diagnosis of Helicobacter pylori infection. METHODS: A total of 205 participants were assessed in Canada and Korea for H. pylori infection status by endoscopy, or a combination of IgG ELISA and CLO test, as well as by the Helikit. The Helikit contains 75 mg of 13C urea as well as citric acid, flavor enhancers and stabilizers in a single plastic cup. The powder is dissolved in 75 mL of water for oral administration. No extra mixing or dilution steps are required. RESULTS: Using the biopsy-derived data as the gold standard the Helikit displayed a clinical sensitivity of 93.5% (95% confidence interval 88.5 to 98.5%) and a clinical specificity of 97.3% (94.3 to 100%). An overall diagnostic efficiency of 95.6% (92.8 to 98.4%) was obtained. No statistically significant difference in the performance characteristics was found between Korea and Canada. No significant adverse events were noted. CONCLUSIONS: The Helikit offers an easy, safe and accurate approach to the diagnosis of H. pylori infection.

Relationship between AgNOR proteins, Ki-67 antigen, p53 immunophenotype and differentiation markers in archival breast carcinomas.

The present study investigated (i) the relationship between standardised morphometric AgNOR parameters (argyrophilic nucleolar organiser region-associated proteins) and MIB1 growth fraction, and (ii) their correlation with immunohistochemical p53, sex steroid receptor status and histopathological differentiation grade in serial paraffin sections from 39 breast carcinomas. Ten sections were double-stained for AgNOR/MIB1. AgNOR parameters correlated significantly with MIB1 growth fraction and p53 protein expression. Significant inverse correlation was found between proliferation markers and oestrogen/progesterone receptor status and histopathological grade. AgNOR expression was significantly higher in cycling (MIB1 positive) tumour cells, than in resting (MIB1 negative) ones, however with exceptions. We conclude, that standardised AgNOR parameters correlate with markers of increased malignant potential in breast carcinomas. However, AgNORs seem to reflect proliferation independent cellular and nucleolar activity of tumour cells, as well. We recommend the use of standardised AgNOR analysis for obtaining sound results in routine paraffin sections.

Evaluation of the i-STAT system: a portable chemistry analyzer for the measurement of sodium, potassium, chloride, urea, glucose, and hematocrit.

OBJECTIVE: To evaluate the analytical performance of the i-STAT system, which is designed for point of care testing and employs a hand-held chemistry analyzer and disposable cartridges, which in the configuration tested, are capable of measuring sodium, potassium, chloride, urea, glucose, and hematocrit in approximately 65 microL of blood in 90's. METHODS: Linearity and imprecision in hematocrit measurement were assessed using whole blood, while that for the other analytes were evaluated with aqueous solutions. The accuracy of the i-STAT system was judged by assay of patient specimens obtained both by venipuncture and fingerprick and correlated with the Kodak Ektachem 700XR and the microhematocrit methods. RESULTS: Linearity was obtained over the clinically relevant range for all analytes. Total imprecision as expressed by the coefficient of variation (CV) was less than 3.5% for all analytes at both high and low concentration except for a low concentration of urea where a CV of 9.4% was obtained. Linear regression analysis revealed minimal systematic errors. The standard error about the regression line (Sy/x) ranged from 0.017 for hematocrit to 2.262 for chloride in the assay of venous blood, whereas in the assay of capillary blood the Sy/x ranged from 0.018 for hematocrit to 0.755 for glucose. CONCLUSIONS: The analytical performance of the i-STAT was deemed acceptable by calculation of total error and comparison with published performance standards. Our study has shown the i-STAT system to be reliable, robust, and simple to operate. Moreover, the compactness of the analyzer and the requirement for only small volumes of whole blood will make it a valuable diagnostic tool in point of care settings.

Effects of epoxyeicosatrienoic acids on isolated hearts and ventricular myocytes.

Effects of cytochrome P-450 metabolites of arachidonic acid, epoxyeicosatrienoic acids (EETS; 5,6-EET, 8,9-EET, 11,12-EET, and 14,15-EET), were examined in isolated guinea pig hearts and ventricular myocytes. Addition of 1-16 ng/ml EETs to normal isolated hearts produced no effects on contractility or coronary pressure. In hearts subjected to 60 min of low-flow ischemia, impairment of contractility and declines in heart rate and coronary perfusion pressure were similar in the presence or absence of 1 ng/ml EETs. However, in the presence of either 5,6- or 11,12-EET, recovery was delayed for the first 10 min only. No significant differences were found in any group regarding heart rate, coronary perfusion pressure, or energy metabolite content after 30 min of reperfusion. In myocytes, both 5,6- and 11,12-EET (100 pg/ml, 1.0 ng/ml, and 20 ng/ml) significantly increased cell shortening as well as intracellular calcium concentrations, whereas 8,9- or 14,15-EET was without effect on these parameters. These results describe for the first time the direct effects of various EETs on cardiac cell function as well as their ability to modulate some of the myocardial responses to postischemic reperfusion. The results suggest a potential role for these substances in the response of the heart to pathological insult.

The catabolism of exogenous lysophosphatidylcholine in isolated perfused rat and guinea pig hearts: a comparative study.

Lysophosphatidylcholine (lysoPC) is an arrhythmogenic phospholipid metabolite which accumulates in the ischemic myocardium. Reduced catabolism of lysoPC has been proposed to be one of the biochemical mechanisms responsible for the increase in lysoPC content. In this investigation we compared the microsomal catabolism of exogenous labeled lysoPC in isolated perfused rat and guinea pig hearts. Analysis of the amount of radioactivity in microsomal phosphatidylcholine (PC) and free fatty acid (FFA) was used as an index of the participation in lysoPC clearance by acylation catalyzed by acyl-CoA:lysoPC acyltransferase and deacylation catalyzed by lysophospholipase, respectively. There was no significant difference in the incorporation of radioactivity into rat and guinea pig heart microsomes; however, the patterns of radioactivity in lysoPC metabolites were notably different. Equal participation by deacylation and reacylation was observed in rat microsomes, whereas deacylation was clearly the preferred route for lysoPC clearance in guinea pig microsomes. Modulation of enzyme activity by treatment of the isolated heart with pHMB, a sulfhydryl agent, was used to probe the relationship among acylation, deacylation and the extent of lysoPC clearance. In guinea pig microsomes impairment of lysoPC acylation was not associated with any change in the amount of radioactivity in lysoPC because of a compensatory increase in deacylation. In contrast, impaired deacylation in rat microsomes led to significant elevations in the amount of radioactivity in lysoPC. We conclude, therefore, that in intact perfused rat and guinea pig hearts the relative participation of acylation and deacylation in lysoPC clearance differs. Moreover, we propose that the level of deacylation by lysophospholipase is an important factor in the extent of clearance of lysoPC.

Effects of palmitoyl carnitine and structural analogs on arterial blood pressure in the rat. A comparison with platelet-activating factor.

Palmitoyl carnitine and lysophosphatidylcholine have been implicated in the generation of cardiac arrhythmias in the ischemic myocardium. These amphiphilic compounds are structurally similar to platelet-activating factor (PAF). The present study compared the hypotensive effect of these compounds to PAF in the anesthetized rat. Palmitoyl carnitine was about 1000 times less potent than PAF in lowering the blood pressure. Lysophosphatidylcholine and other structurally related compounds, in dosages similar to that of palmitoyl carnitine, had no significant hypotensive action. CV 3988, a PAF antagonist, blocked the hypotensive action of PAF but had no effect on the hypotensive action of palmitoyl carnitine. This suggested the effect of palmitoyl carnitine was not associated with the same site or mechanism as PAF. The results also ruled out the involvement of prostaglandin formation and of the sympathetic nervous system since indomethacin, phenoxybenzamine and propranolol did not affect the hypotensive action of palmitoyl carnitine. In addition, it is unlikely that palmitoyl carnitine exerted its effect by a direct membrane-perturbing action because lysophosphatidylcholine, which possesses similar amphiphilic properties, does not share the same hypotensive effect.

Mechanism of lysophosphatidylcholine accumulation in the ischemic canine heart.

The metabolism of lysophosphatidylcholine (LPC) in non-ischemic and ischemic canine heart was investigated by in vitro enzyme analysis. Selected subcellular fractions were assayed for the LPC-producing enzyme phospholipase A and the LPC-eliminating enzymes LPC:acyl-CoA acyltransferase, LPC:LPC transacylase and lysophospholipase. The canine heart was found to contain all enzymes differing, however, in subcellular distribution and specific activity. Phospholipase A activity did not change significantly in any of the fractions prepared from the ischemic tissue of hearts rendered ischemic for 1, 3 or 5 hr when compared to non-ischemic tissue. Changes in the activity of the microsomal LPC:acyl-CoA acyltransferase over the course of 5 hr of ischemia were observed. Significant decreases in the activity of the cytosolic and microsomal lysophospholipases were detected especially after 3 and 5 hr of ischemia. Similarly, a decrease in the activity of the microsomal LPC:LPC transacylase was noted after 3 and 5 hr of ischemia. Our results suggest that impaired catabolism of LPC rather than an enhanced production of LPC is the principal mechanism for the increase in LPC levels in the ischemic canine heart.

The catabolism of plasmenylcholine in the guinea pig heart.

The hydrolysis of the alkenyl bonds of plasmenylcholine and plasmenylethanolamine by plasmalogenase, followed by hydrolysis of the resultant lysophospholipid by lysophospholipase, has been postulated as the major pathway for the catabolism of these plasmalogens. However, the postulation was based solely on the presence of plasmalogenase activity towards plasmenylethanolamine and plasmenylcholine in the brain. In this study we have demonstrated the absence of plasmalogenase activity for plasmenylcholine in the guinea pig heart under a wide range of experimental conditions. Plasmenylcholine was hydrolysed by phospolipase A2 activities in cardiac microsomal, mitochondrial and cytosolic fractions. Phospholipase A2 activities in these fractions had an alkaline pH optimum and were enhanced by Ca2+. The enzymes also displayed high specificity for plasmenylcholine with linoleoyl or oleoyl at the C-2 position. Lysoplasmalogenase activity for lysoplasmenycholine was also detected and characterized in the microsomal and mitochondrial fractions. Since the cardiac plasmalogenase is only active towards plasmenylethanolamine but not plasmenylcholine, the catabolism of these two plasmalogens must be different from each other. We postulate that the major pathway for the catabolism of plasmenycholine involves the hydrolysis of the C-2 fatty acid by phospholipase A2, and hydrolysis of the vinyl ether group of the resultant lysoplasmenylcholine by lysoplasmalogenase.

Effects of fatty acids on phosphatidylcholine biosynthesis in isolated hamster heart.

The effects of stearic, oleic, and arachidonic acids on phosphatidylcholine biosynthesis in the hamster heart were investigated. When hamster hearts were perfused with labelled choline in the presence of fatty acids, biosynthesis of phosphatidylcholine was stimulated only by stearic acid. Stearic acid was found to accumulate in unesterified (free) form in the hamster heart after perfusion. The stimulation by stearic acid was mediated in vivo by an enhancement of CTP:phosphocholine cytidylyltransferase activity in the microsomal fraction of the hamster heart and the enzyme activity in the cytosolic fraction was not affected. In contrast with the observations in rat hepatocytes, cytidylyltransferase from the hamster heart was not stimulated directly by stearic acid. The selective activation of the microsomal enzyme when the heart was perfused with stearic acid suggests that activation of the enzyme was mediated via the modification of the membrane by stearic acid.

Detection of heterotopic gray matter in children by magnetic resonance imaging.

Heterotopic gray matter results from abnormal brain development and is a recognized focus of seizures. It may be associated with mental retardation and/or severe malformations of the brain. Three patients with heterotopia of gray matter were identified by magnetic resonance imaging (MRI). CT failed to detect the heterotopic gray matter in each case. One child was referred for removal of a neoplasm based on CT studies until MRI demonstrated the developmental nature of his condition. One infant had severely dysplastic left cerebral hemisphere associated with heterotopic gray matter and the syndrome of Hypomelanosis of Ito. All three children suffered from seizures and/or mental retardation. MRI provided important information in the management of each case and appears to be the imaging method of choice in evaluating children with seizures or retardation for heterotopic gray matter in the brain.

The distribution and acyl composition of plasmalogens in guinea pig heart.

In guinea pig heart homogenate, 34% of both choline and ethanolamine phosphoglycerides were in the form of plasmalogens (1-alkenyl, 2-acyl glycerophospholipid). Plasmalogens accounted for 39% of the choline phosphoglycerides and 36% of the ethanolamine phosphoglycerides in the mitochondrial fraction. Ethanolamine plasmalogen was the major ethanolamine phosphoglyceride (63%) in the guinea pig heart microsomal fraction. A high arachidonyl content was found in both diacyl and 1-alkenyl, 2-acyl glycerophosphoethanolamine. The C-2 fatty acyl profiles of the diacyl and 1-alkenyl, 2-acyl choline phosphoglycerides differed considerably from each other in the homogenate as well as in the subcellular fractions. Significant differences in the C-2 fatty acyl profiles also were observed in diacyl and 1-alkenyl, 2-acyl ethanolamine phosphoglycerides. Such differences suggest there is no direct metabolic relationship between the diacyl glycerophosphocholine (-ethanolamine) and its plasmalogen analog.

Alterations in lysophosphatidylcholine levels of canine heart: modes of extraction and storage.

The extraction of lysophosphatidylcholine from canine heart and the storage of lysophosphatidylcholine in total lipid extracts were investigated. The lysolipid was effectively extracted from canine heart by chloroform/methanol (1/2) and the maximum recovery of the lysolipid was 89-92%. Changes in levels of the lysolipid were observed when the tissue was stored at 0 degree C for 60 min or at -20 degrees C for 7 days. An alteration in the lysophosphatidylcholine level was also observed when the lipid extract from the heart was stored in theoretical lower phase (chloroform/methanol/water; 86/14/1). In order to accurately assess the level of lysophosphatidylcholine in the canine heart, the lipid should be extracted immediately from fresh tissue and the lipid extract stored in chloroform/methanol (2/1) or without solvent under nitrogen prior to separation and determination.