Vasoproliferative process resembling pulmonary capillary hemangiomatosis in a cat.

BACKGROUND: Pulmonary capillary hemangiomatosis is a rare, vascular obstructive disorder that uniformly causes pulmonary arterial hypertension. Clinically, pulmonary capillary hemangiomatosis is indistinguishable from primary pulmonary arterial hypertension and histology is required for definitive diagnosis. The distinctive histologic feature of pulmonary capillary hemangiomatosis is non-malignant extensive proliferation of capillaries in the alveolar septae. Vasodilator treatment of humans with primary arterial hypertension due to pulmonary capillary hemangiomatosis can result in fatal acute pulmonary edema. Computed tomography is thus critical to discern pulmonary capillary hemangiomatosis from other causes of pulmonary arterial hypertension prior to vasodilator therapy. This is the first report of a vasoproliferative process resembling pulmonary capillary hemangiomatosis in the feline species. CASE PRESENTATION: A 15-year-old, male castrated, domestic shorthair cat presented for persistent labored breathing presumptively due to congestive heart failure despite treatment with diuretics for 7 days. Echocardiography showed evidence of hypertrophic cardiomyopathy with severe pulmonary hypertension; however, a normal sized left atrium was not consistent with congestive heart failure. Thoracic computed tomography was performed and showed evidence of diffuse ill-defined nodular ground glass opacities, enlarged pulmonary arteries, and filling defects consistent with pulmonary thromboembolism. The cat acutely decompensated after a single dose of sildenafil and was euthanized. Histopathology of the lungs showed severe multifocal alveolar capillary proliferation with respiratory bronchiolar infiltration, marked type II pneumocyte hyperplasia and multifocal pulmonary arterial thrombosis. CONCLUSION: This is the first description in a cat of a vasoproliferative disorder resembling pulmonary capillary hemangiomatosis complicated by multifocal pulmonary arterial thrombosis. Inspiratory and expiratory ventilator-driven breath holds with angiography revealed lesions predominantly characterized by ground glass opacification and vascular filling defects with absence of air trapping. The results from this report suggest that, as in humans, the cat can develop a pulmonary capillary hemangiomatosis-like disease in which vasodilator therapy to address pulmonary hypertension may lead to fatal pulmonary edema.

Exposure to a combination of heat and hyperoxia during cycling at submaximal intensity does not alter thermoregulatory responses.

In this study, we tested the hypothesis that breathing hyperoxic air (FinO2 = 0.40) while exercising in a hot environment exerts negative effects on the total tissue level of haemoglobin concentration (tHb); core (Tcore) and skin (Tskin) temperatures; muscle activity; heart rate; blood concentration of lactate; pH; partial pressure of oxygen (PaO2) and carbon dioxide; arterial oxygen saturation (SaO2); and perceptual responses. Ten well-trained male athletes cycled at submaximal intensity at 21°C or 33°C in randomized order: first for 20 min while breathing normal air (FinO2 = 0.21) and then 10 min with FinO2 = 0.40 (HOX). At both temperatures, SaO2 and PaO2, but not tHb, were increased by HOX. Tskin and perception of exertion and thermal discomfort were higher at 33°C than 21°C (p < 0.01), but independent of FinO2. Tcore and muscle activity were the same under all conditions (p > 0.07). Blood lactate and heart rate were higher at 33°C than 21°C. In conclusion, during 30 min of submaximal cycling at 21°C or 33°C, Tcore, Tskin and Tbody, tHb, muscle activity and ratings of perceived exertion and thermal discomfort were the same under normoxic and hyperoxic conditions. Accordingly, breathing hyperoxic air (FinO2 = 0.40) did not affect thermoregulation under these conditions.

Influence of the gut microbiota on blood acute-phase proteins.

Little is known about the bovine intestinal microbiota influence on systemic innate immune responses. The objective of the present study was to determine relationships between acute-phase proteins in blood serum of cows [C-reactive protein (CRP), LPS-binding protein (LBP) and haptoglobin (Hp)] and the faecal microbiota. Fifty-two healthy cows (2-8 years old) were investigated. Faecal bacteria were determent characterized by in situ hybridization with 16S/23S rRNA-targeted probes and by conventional culture methods. The population of Gram-negative faecal bacteria (Enterobacteriaceae) was correlated negatively with CRP and positively with LBP in blood plasma, independent of the method used. Similar results were observed with Clostridium perfringens. No correlation was found between the faecal population of intestinal bacteria and Hp levels in blood plasma. This datum indicates that intestinal bacteria, especially Enterobacteriaceae and C. perfringens, may influence the level of CRP and LBP in blood plasma. These findings can be very important for diagnostic evaluations of the intestinal microbiota and provide specific information about its regulation.

Cardiorespiratory, metabolic and hormonal responses during open-wheel indoor kart racing.

AIM: This study aimed to quantify the cardiorespiratory, metabolic and hormonal responses of elite open-wheel indoor kart racers. METHODS: Ten male racers (age: 21±3 yrs; height: 1.92±0.06 m, body mass: 76.0±5.9 kg) participated in a racing tournament. Their peak oxygen uptake and heart rate were assessed by a ramp test (100 W, increase 30 W·min-1) in the laboratory. During the racing itself, the cardio-respiratory and accelerometer values were recorded and pre- and post-race levels of blood lactate and salivary cortisol were determined. RESULTS: The average peak values for all of the drivers with respect to oxygen uptake and heart rate were 4.5±0.8 L·min-1 (56.7±7.9 mL·min-1·kg-1) and 193±5 beats·min-1, respectively. Overall, 28.3±3.3 laps were completed during 30-min of racing. Acceleration forces for the entire test averaged 1.20±0.51 G (maximum: 3.30 G), declining from the first 10 min until the end of racing (P<0.03). The oxygen uptake (~20 mL·min-1·kg-1), heart rate (~133 beats·min-1), respiratory exchange ratio (~0.96) and ventilation (~70 L·min-1) observed indicated moderate cardio-respiratory responses. Blood lactate concentration was significantly higher after the race than before but remained at <2 mmol·L-1 (P<0.01; effect size: 1.62). CONCLUSION: There were no differences between salivary cortisol levels before and after the race (P<0.06; effect size: 0.49). Directly after the race, the drivers rated their perceived exertion on Borg's scale as 11.1±1.3. The present data revealed that the psycho-physical exertion associated with a 30-min open-wheel indoor kart race is moderate.

Bioluminescence assay for the highly sensitive detection of botulinum neurotoxin A activity.

This article describes a novel bioluminescence assay for detecting the proteolytic activity of Botulinum NeuroToxins (BoNT) in complex matrices. The assay is capable of detecting traces of BoNT in blood samples as well as in food drinks. The assay was responsive to BoNT/A subtypes 1 to 5, and serotype E3 in buffered solutions. It was responsive to filtered Clostridium botulinum supernatants and BoNT/A1 in complex with neurotoxin associated proteins in bouillon and milk (3.8% fat) down to 400 fM after 4 h RT incubation and in bouillon at concentrations down to 120 fM after 21 h RT incubation. In combination with an immunocapture/enrichment step it could detect BoNT/A1 in citrated plasma at concentrations down to 30 fM (1.2 mouse LD50 per mL). The simplicity of the assay, combined with a demonstrated ability to lyophilize the reagents, demonstrates its usefulness for detection of BoNT in non-specialised analytical laboratories.

Active sulfur cycling by diverse mesophilic and thermophilic microorganisms in terrestrial mud volcanoes of Azerbaijan.

Terrestrial mud volcanoes (TMVs) represent geochemically diverse habitats with varying sulfur sources and yet sulfur cycling in these environments remains largely unexplored. Here we characterized the sulfur-metabolizing microorganisms and activity in four TMVs in Azerbaijan. A combination of geochemical analyses, biological rate measurements and molecular diversity surveys (targeting metabolic genes aprA and dsrA and SSU ribosomal RNA) supported the presence of active sulfur-oxidizing and sulfate-reducing guilds in all four TMVs across a range of physiochemical conditions, with diversity of these guilds being unique to each TMV. The TMVs varied in potential sulfate reduction rates (SRR) by up to four orders of magnitude with highest SRR observed in sediments where in situ sulfate concentrations were highest. Maximum temperatures at which SRR were measured was 60°C in two TMVs. Corresponding with these trends in SRR, members of the potentially thermophilic, spore-forming, Desulfotomaculum were detected in these TMVs by targeted 16S rRNA analysis. Additional sulfate-reducing bacterial lineages included members of the Desulfobacteraceae and Desulfobulbaceae detected by aprA and dsrA analyses and likely contributing to the mesophilic SRR measured. Phylotypes affiliated with sulfide-oxidizing Gamma- and Betaproteobacteria were abundant in aprA libraries from low sulfate TMVs, while the highest sulfate TMV harboured 16S rRNA phylotypes associated with sulfur-oxidizing Epsilonproteobacteria. Altogether, the biogeochemical and microbiological data indicate these unique terrestrial habitats support diverse active sulfur-cycling microorganisms reflecting the in situ geochemical environment.

Prolongation of electrode lifetime in biofuel cells by periodic enzyme renewal.

Enzymatically catalyzed biofuel cells show unique specificity and promise high power densities, but suffer from a limited lifetime due to enzyme deactivation. In the present work, we demonstrate a novel concept to extend the lifetime of a laccase-catalyzed oxygen reduction cathode in which we decouple the electrode lifetime from the limited enzyme lifetime by a regular resupply of fresh enzymes. Thereto, the adsorption behavior of laccase from Trametes versicolor to buckypaper electrode material, as well as its time-dependent deactivation characteristics, has been investigated. Laccase shows a Langmuir-type adsorption to the carbon nanotube-based buckypaper electrodes, with a mean residence time of 2 days per molecule. In a citrate buffer of pH 5, laccase does not show any deactivation at room temperature for 2 days and exhibits a half-life of 9 days. In a long-term experiment, the laccase electrodes were operated at a constant galvanostatic load. The laccase-containing catholyte was periodically exchanged against a freshly prepared one every second day to provide sufficient active enzymes in the catholyte for the replacement of desorbed inactive enzymes. Compared to a corresponding control experiment without catholyte exchange, this procedure resulted in a 2.5 times longer cathode lifetime of 19 ± 9 days in which the electrode showed a potential above 0.744 V vs. normal hydrogen electrode at 110 µA cm(-2). This clearly indicates the successful exchange of molecules by desorption and re-adsorption and is a first step toward the realization of a self-regenerating enzymatic biofuel cell in which enzyme-producing microorganisms are integrated into the electrode to continuously resupply fresh enzymes.

Ergogenic effect of hyperoxic recovery in elite swimmers performing high-intensity intervals.

This investigation tested the hypothesis that breathing oxygen-enriched air (F(i)O(2) =1.00) during recovery enhances peak (P(peak)) and mean power (P(mean)) output during repeated high-intensity exercise. Twelve elite male swimmers (21 ± 3 years, 192.1 ± 5.9 cm, 79.1 ± 8.2 kg) inhaled either hyperoxic (HOX) or normoxic (NOX) air during 6-min recovery periods between five repetitions of high-intensity bench swimming, each involving 40 maximal armstrokes. Oxygen partial pressure (pO(2)) and saturation (SO(2)), [H(+)], pH, base excess and blood lactate concentration were measured before and after all intervals. The production of the reactive oxygen species (ROS) hydrogen peroxide was measured before, directly after and 15 min after the test. P(peak) and P(mean) with HOX recovery were significantly higher than with NOX throughout the third, fourth and fifth intervals (P<0.001-0.04). With HOX, electromyography activity was lower during the third, fourth and fifth intervals than during the first (P=0.05-0.001), with no such changes in NOX (P=0.99). There were no differences in blood lactate, pH, [H(+)] or base excess and ROS production at any time point with either HOX or NOX recovery. These findings demonstrate that the P(peak) and P(mean) of elite swimmers performing high-intensity intervals can be improved by exposure to oxygen-enriched air during recovery.

Effect of a Potential Probiotic Candidate Enterococcus faecalis-1 on Growth Performance, Intestinal Microbiota, and Immune Response of Commercial Broiler Chickens.

The probiotic effect of Enterococcus faecalis-1 (isolated from healthy chickens) on growth performance, immune response, and modulation of the intestinal microbiota of broilers was assessed with a total of 100-day-old commercial Cobb chicks. The chicks were randomly divided into two equal groups. The control group received a basal diet, while the test group received a basal diet and was orally supplied with E. faecalis at a dose of 108 CFU/bird/day. Results showed that E. faecalis-1 supplement significantly (P < 0.05) improved the body weight and feed conversion ratio of treated broilers compared with the control ones. The mortality percentage was reduced in E. faecalis-1-supplemented group. The total IgY serum level was significantly (P < 0.05) increased in broilers receiving E. faecalis-1 supplement (7.1 ± 0.39) compared with the control group (5.8 ± 0.3), while the serum avidin level was significantly (P < 0.05) decreased in E. faecalis-1-supplemented broilers (76 ± 11.1). There was no significant change in the immune response towards avian influenza and Newcastle vaccines in both groups. The total Lactobacillus and Enterococcus counts were significantly (P < 0.05) higher in the cecal contents of broilers given E. faecalis-1 than those that received the control treatment. E. faecalis-1 supplement enhanced the enzyme activities, antioxidant system, and liver functions of treated broilers compared with those in the control group. Collectively, these results showed that E. faecalis-1 could promote growth performance and immunological status and convey beneficial modulation of the cecal microbiota in broilers.

Decreased microglial numbers in Vav1-Cre+:dicer knock-out mice suggest a second source of microglia beyond yolk sac macrophages.

Microglia represent the resident macrophages of the central nervous system (CNS). While it is clear that microglia recruitment is established by differentiation of primitive yolk sac (YS) macrophages and consecutive invasion of the brain, starting around E8 in rodents (Ginhoux et al., 2010), more recent studies suggest that a non-YS contribution to the microglia population should not entirely be dismissed (Swinnen et al., 2013; Xu et al., 2015). Therefore, we used Vav1-Cre+:dicer knock-out mice in order to study the effect of the post-YS hematopoiesis on the definitive microglial population in late prenatal (E16.5, E18.5) and early postnatal brains (P0, P1). Since Vav1 is thereby exclusively expressed in hematopoietic cells starting at E11, the depletion of the micro RNA processing enzyme dicer in Vav1-positive cells allows interfering with post-YS microglia recruitment. Using this approach, analysis of the number of Iba-1 positive microglia revealed a reduction of microglial numbers by 40% in knock-out mice at P1 compared to their individual control littermates. Noteworthy, immunolabeling for Ki-67 and active caspase 3 confirmed that the differences in the microglial numbers are not related to differential rates of proliferation or apoptosis. Therefore, our data demonstrates that interfering with the definitive hematopoiesis highly impacts on the microglial population, implicating an important role of post-YS hematopoiesis on microglial development and recruitment.

Similar precipitated withdrawal effects on intracranial self-stimulation during chronic infusion of an e-cigarette liquid or nicotine alone.

The FDA recently extended their regulatory authority to electronic cigarettes (ECs). Because the abuse liability of ECs is a leading concern of the FDA, animal models are urgently needed to identify factors that influence the relative abuse liability of these products. The ability of tobacco products to induce nicotine dependence, defined by the emergence of anhedonia and other symptoms of nicotine withdrawal following cessation of their use, contributes to tobacco abuse liability. The present study compared the severity of precipitated withdrawal during chronic infusion of nicotine alone or nicotine-dose equivalent concentrations of three different EC refill liquids in rats, as indicated by elevations in intracranial self-stimulation (ICSS) thresholds (anhedonia-like behavior). Because these EC liquids contain constituents that may enhance their abuse liability (e.g., minor alkaloids), we hypothesized that they would be associated with greater withdrawal effects than nicotine alone. Results indicated that the nicotinic acetylcholine receptor antagonist mecamylamine precipitated elevations in ICSS thresholds in rats receiving a chronic infusion of nicotine alone or EC liquids (3.2mg/kg/day, via osmotic pump). Magnitude of this effect did not differ between formulations. Our findings indicate that nicotine alone is the primary CNS determinant of the ability of ECs to engender dependence. Combined with our previous findings that nicotine alone and these EC liquids do not differ in other preclinical addiction models, these data suggest that product standards set by the FDA to reduce EC abuse liability should primarily target nicotine, other constituents with peripheral sensory effects (e.g. flavorants), and factors that influence product appeal (e.g., marketing).

Identification of RIP1 as a critical mediator of Smac mimetic-mediated sensitization of glioblastoma cells for Drozitumab-induced apoptosis.

This study aims at evaluating the combination of the tumor-necrosis-factor-related apoptosis-inducing ligand (TRAIL)-receptor 2 (TRAIL-R2)-specific antibody Drozitumab and the Smac mimetic BV6 in preclinical glioblastoma models. To this end, the effect of BV6 and/or Drozitumab on apoptosis induction and signaling pathways was analyzed in glioblastoma cell lines, primary glioblastoma cultures and glioblastoma stem-like cells. Here, we report that BV6 and Drozitumab synergistically induce apoptosis and reduce colony formation in several glioblastoma cell lines (combination index<0.1). Also, BV6 profoundly enhances Drozitumab-induced apoptosis in primary glioblastoma cultures and glioblastoma stem-like cells. Importantly, BV6 cooperates with Drozitumab to suppress tumor growth in two glioblastoma in vivo models including an orthotopic, intracranial mouse model, underlining the clinical relevance of these findings. Mechanistic studies reveal that BV6 and Drozitumab act in concert to trigger the formation of a cytosolic receptor-interacting protein (RIP) 1/Fas-associated via death domain (FADD)/caspase-8-containing complex and subsequent activation of caspase-8 and -3. BV6- and Drozitumab-induced apoptosis is blocked by the caspase inhibitor zVAD.fmk, pointing to caspase-dependent apoptosis. RNA interference-mediated silencing of RIP1 almost completely abolishes the BV6-conferred sensitization to Drozitumab-induced apoptosis, indicating that the synergism critically depends on RIP1 expression. In contrast, both necrostatin-1, a RIP1 kinase inhibitor, and Enbrel, a TNFα-blocking antibody, do not interfere with BV6/Drozitumab-induced apoptosis, demonstrating that apoptosis occurs independently of RIP1 kinase activity or an autocrine TNFα loop. In conclusion, the rational combination of BV6 and Drozitumab presents a promising approach to trigger apoptosis in glioblastoma, which warrants further investigation.

A novel acute phase marker in cattle: lipopolysaccharide binding protein (LBP).

The host response to infection, the "acute phase response" is a highly conserved series of physiological reactions including marked changes in concentrations of plasma proteins. These proteins have been shown to participate in the immune response to infections. Several recent studies have elevated the role of acute phase proteins (APPs) as predictive markers in infection. APPs such as serum amyloid A and haptoglobin but not C-reactive protein (CRP) have been identified as markers of inflammation in cattle. In humans, lipopolysaccharide (LPS) binding protein (LBP) has certain biological functions in host defence and participates in acute phase reactions. We measured plasma levels of LBP in a group of 20 calves experimentally infected with Gram-negative Mannheimia haemolytica (Pasteurella) in comparison to haptoglobin, the most widely studied APP in cattle. In infected calves, LBP levels rose significantly 6 h after infection, reaching a maximum at 24 h. Haptoglobin concentrations significantly rose after 12 h, and peak responses were measured 48 h after infection. Thus, LBP may prove to be a diagnostic marker in cattle infection and is faster than haptoglobin in detecting sepsis.

Molecular size and flexibility as determinants of selectivity in the oxidation of N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine analogs by monoamine oxidase A and B.

The introduction of a methylene bridge between the phenyl and tetrahydropyridyl moieties of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) results in increased selectivity for monoamine oxidase B (MAO B) over monoamine oxidase A (MAO A). However, lengthening of this bridge results in a total loss of selectivity. In the present study, a number of isomeric 4-naphthyl-, 4-(naphthylalkyl)-, 4-thienyl-, and 4-(thienylalkyl)tetrahydropyridines, conformationally restrained and flexible analogs of MPTP, were synthesized and evaluated as potential selective substrates of MAO A and B. In terms of the parameter (turnover number)/Km, the bulky naphthyl analogs were invariably better substrates of MAO A than kynuramine, the reference substrate for this enzyme. In addition, all naphthyl analogs, regardless of conformational mobility, were more effective substrates of MAO A than MAO B. Similarly, all thienyl analogs were found to be more effective substrates of MAO B. In contrast to the naphthalenes, the conformationally restrained thiophenes 9a and 10a were found to be poor substrates of MAO B, relative to benzylamine, the reference substrate. These results suggest that the selectivity of these compounds for either MAO A or B is determined by the complex interplay of molecular size and flexibility. In this interplay, either one of these two factors may predominate.

Interaction of tetrahydrostilbazoles with monoamine oxidase A and B.

1-Methyl-1,2,3,6-tetrahydrostilbazole (MTHS) and its analogs are oxidized by monoamine oxidase (MAO) A at slow rates comparable to that for the structurally similar neurotoxin, 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, but the rates of oxidation by MAO B vary over a wide range depending on the structure of the analog. MAO A oxidation of all of the analogs yielded nonhyperbolic kinetic patterns, with little difference between the cis and trans isomers. In contrast MAO B showed hyperbolic kinetics and distinct stereoselectivity for the cis isomers. The corresponding pyridinium forms of trans-MTHS and its analogs were more potent inhibitors of MAO A (Ki values between 0.3 and 5 microM) than of MAO B, for which the Ki values varied greatly. The data suggest that the stringency of the MAO A active site for the geometry of the substrate molecule is less strict than that of MAO B. With MAO B, any substitution on the phenyl ring can lead to dramatic changes in the substrate properties which may be explained by the different orientation of substrate at the active site of the enzyme. Molecular geometry but not the effects of the substituents was shown to be an important factor in determining the effectiveness of substrate oxidation by MAO B.

Quantitative light and electron microscopic changes in thymic reticular epithelial cells during Moloney-virus-induced lymphoma development.

This report describes two types of reticular epithelial cell in the thymic cortex of the BALB/c mouse, an immature and a mature form. During early stages of lymphoma development, i.e., 2-6 weeks postinfection (p.i.) with Moloney leukemia virus (M-MuLV), activation of the epithelial cells is observed. Although the percentage of these cells in the total cell population of the thymic cortex remains constant during that time, the number of mature epithelial cells is significantly increased in infected animals. Subsequently, about 6 weeks p.i., the number of immature epithelial cells starts to increase, whereas the number of mature reticular epithelial cells declines and the appearance of the mature epithelial cells changes drastically. The results of light and electron microscopic studies indicate degeneration of the mature reticular epithelial cells at the onset of lymphoma development at a time when the first deficiencies in the immunologic competence of the reticular epithelial cells are apparent.