Pharmacokinetics of furosemide administered 4 and 24 hours prior to high-speed exercise in horses.

Furosemide is a diuretic agent used commonly in racehorses to attenuate the bleeding associated with exercise-induced pulmonary hemorrhage (EIPH). The current study describes serum and urine concentrations and the pharmacokinetics of furosemide following administration at 4 and 24 hrs prior to maximal exercise. Eight exercised adult Thoroughbred horses received a single IV administration of 250 mg of furosemide at 4 and 24 hrs prior to maximal exercise on a high-speed treadmill. Blood and urine samples were collected at time 0 and at various times for up to 72 hrs and furosemide concentrations determined using liquid chromatography-tandem mass spectrometry. Serum furosemide concentrations remained above the LOQ (0.05 ng/ml) for 36 hrs in 3/8 and 1/8 horses in the 4- and 24-hrs groups, respectively. Serum concentration data were best fit by a two-compartment model. There was not a significant difference in the volume of distribution at steady-state (0.594 ± 0.178 [4 hrs] and 0.648 ± 0.147 [24 hrs] L/kg) or systemic clearance (0.541 ± 0.094 [4 hrs] and 0.617 ± 0.114 [24 hrs] L/hrs/kg) between horses that were exercised at 4- and 24 hrs postdrug administration. The mean ± SD elimination half-life was 3.12 ± 0.387 and 3.23 ± 0.407 hrs following administration at 4 and 24 hrs prior to exercise, respectively.

A multinational survey of companion animal veterinary clinicians: How can antimicrobial stewardship guidelines be optimised for the target stakeholder?

Antimicrobial stewardship initiatives are widely regarded as a cornerstone for ameliorating the global health impact of antimicrobial resistance. Within companion animal health, such efforts have largely focused on development and dissemination of antimicrobial stewardship guidelines (ASGs). However, there have been few attempts to understand veterinarian attitudes towards and knowledge of ASGs or to determine how awareness regarding ASGs might best be increased. An online survey regarding ASGs was formulated for veterinarians who treat companion animals. The survey was distributed across 46 European and associated countries between 12 January and 30 June, 2022. In total, 2271 surveys were completed, with 64.9% of respondents (n = 1474) reporting awareness and usage of at least one ASG. Respondents from countries with greater awareness of ASGs tended to report more appropriate use of antimicrobials (Spearman's rank coefficient = 0.6084, P ≤ 0.001), with respondents from countries with country-specific ASGs tending to score highest across both awareness and appropriate use domains. Respondents prioritised guidance around antimicrobial choice (82.0%, n = 1863), duration of treatment (66.0%, n = 1499), and dosage (51.9%, n = 1179) for inclusion in future ASGs, with 78.0% (n = 1776) of respondents preferring ASGs to be integrated into their patient management system. Awareness of ASGs and their use in companion animal veterinary practice appears to be greater than previously reported, with respondents tending to report antimicrobial prescription decision making broadly in line with current clinical recommendations. However, further initiatives aimed at maximising accessibility to ASGs both within countries and individual veterinary practices are recommended.

Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a specific substrate of yeast metacaspase.

Yeast metacaspase (Yca1p) is required for the execution of apoptosis upon a wide range of stimuli. However, the specific degradome of this yeast protease has not been unraveled so far. By combining different methodologies described as requisites for a protein to be considered a protease substrate, such as digestome analysis, cleavage of recombinant GAPDH by metacaspase and evaluation of protein levels in vivo, we show that upon H(2)O(2)-induced apoptosis, the metabolic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is a specific target of metacaspase. Nitric oxide (NO) signaling, which mediates H(2)O(2)-induced apoptosis, is required for metacaspase specific GAPDH cleavage. In conclusion, in this work we identified GAPDH as the first direct yeast metacaspase substrate described so far. Although mammalian caspases and yeast metacaspase apparently have distinct target cleavage sites, GAPDH arises as a common substrate for these proteases.

Proposal of a histopathological predictive rule for the differential diagnosis between American tegumentary leishmaniasis and sporotrichosis skin lesions.

BACKGROUND: American tegumentary leishmaniasis (ATL) and sporotrichosis exhibit similar histopathology and low frequencies of microorganism detection. OBJECTIVES: This study seeks to identify microscopic alterations that can distinguish between these diseases. METHODS: Haematoxylin and eosin stained slides of 171 ATL and 97 sporotrichosis samples from active cutaneous lesions were examined for histopathological alterations. The lesions were diagnosed by isolating the agent (which was not visible) in culture. An intuitive diagnosis was assigned to each slide. The strength of the association between the histopathological findings and the diagnosis was estimated by an odds ratio, and each finding was graded according to a regression model. A score was assigned to each sample based on the histopathological findings. A study of the interobserver reliability was performed by calculating kappa coefficients of the histopathological findings and intuitive diagnoses. RESULTS: The markers 'macrophage concentration', 'tuberculoid granuloma' and 'extracellular matrix degeneration' were associated with ATL. 'Suppurative granuloma', 'stellate granuloma', 'different types of giant cells', 'granulomas in granulation tissue' and 'abscess outside the granuloma' were associated with a diagnosis of sporotrichosis. 'Macrophage concentration' and 'suppurative granuloma' had the highest (substantial and almost perfect, respectively) reliability. The regression model score indicated 92.0% accuracy. The intuitive diagnosis had 82.5% diagnostic accuracy and substantial reliability. CONCLUSIONS: Taking into account the clinical and epidemiological context, some histopathological alterations might be useful for the differential diagnosis between ATL and sporotrichosis cutaneous lesions in cases in which the aetiological agent is not visible.

Pharmacokinetics and preliminary safety evaluation of azithromycin in adult horses.

Azithromycin is widely used in foals but has not been studied in adult horses. The goals of this study were to determine the pharmacokinetic profile and to make a preliminary assessment of the safety of azithromycin in adult horses. Azithromycin was administered intravenously (5 mg/kg) and intragastrically (10 mg/kg) to six healthy mares in a crossover design. Serial plasma samples, blood neutrophils, and pulmonary macrophages were collected for the measurement of azithromycin concentrations. Azithromycin was also administered orally (10 mg/kg) once a day for 5 days to five healthy mares for preliminary evaluation of safety in adult horses. The bioavailability of azithromycin following intragastric administration was 45 ± 12%. Concentrations within peripheral neutrophils and bronchoalveolar macrophages were several fold higher than that of plasma. Mild decreases in appetite (n = 3) and alterations in fecal consistency (n = 3) were noted following repeated oral administration. The pharmacokinetic profiles of azithromycin in adult horses, especially the slow elimination rate and intraneutrophil and intrapulmonary macrophage accumulation, demonstrate that it is conducive to use in this age group. Because of the gastrointestinal alterations noted, further studies are warranted before azithromycin can be recommended for use in adult horses.

Adhesive and biodegradable membranes made of sustainable catechol-functionalized marine collagen and chitosan.

We describe bioadhesive membranes developed from marine renewable biomaterials, namely chitosan and collagen extracted from fish skins. Collagen was functionalized with catechol groups (Coll-Cat) to provide the membranes with superior adhesive properties in a wet environment and blended with chitosan to improve the mechanical properties. The blended membranes were compared to chitosan and chitosan blended with unmodified collagen in terms of surface morphology, wettability, weight loss, water uptake, mechanical and adhesive properties. The metabolic activity, the viability and the morphology of L929 fibroblastic cells seeded on these membranes were also assessed. Our results show that the functionalization with catechol groups improves the adhesive and mechanical properties of the membranes and enhances cell attachment and proliferation. These data suggest that the developed marine origin-raw membranes present a potential towards the restoration of the structural and functional properties of damaged soft tissues.

Riboflavin did not provide anti-inflammatory or antioxidant effects in an experimental model of sepsis.

We aimed to evaluate whether the administration of riboflavin to septic animals reduces inflammation, oxidative stress, organ dysfunction, and mortality. C57BL/6 mice, 6-8 weeks old, were allocated to the study group (polymicrobial sepsis induced by cecal ligation and puncture (CLP) + antibiotic + iv riboflavin), control (CLP + antibiotic + iv saline), or naïve (non-operated controls). Serum concentrations of alanine aminotransferase (ALT), creatine kinase-MB (CK-MB), urea, and creatinine, and markers of inflammation [interleukin (IL)-6, tumor necrosis factor (TNF)-α, keratinocyte-derived chemokine (KC), and macrophage inflammatory protein (MIP)-2)], and oxidative stress (malondialdehyde (MDA) were measured 12 h after the experiment. Animal survival rates were calculated after 7 days. Means between groups were compared using linear regression models adjusted under the Bayesian approach. No significant difference was observed between control and study groups in serum concentrations of IL-6 (95% credible interval) (-0.35 to 0.44), TNF-α (-15.7 to 99.1), KC (-0.13 to 0.05), MIP-2 (-0.84 to 0.06), MDA (-1.25 to 2.53), or ALT (-6.6 to 11.5). Serum concentrations of CK-MB (-145.1 to -30.1), urea (-114.7 to -15.1), and creatinine (-1.14 to -0.01) were higher in the study group. Survival was similar in both groups (P=0.8). Therefore, the use of riboflavin in mice undergoing sepsis induced by CLP did not reduce inflammation, oxidative stress, organ dysfunction, or mortality compared with placebo.

ITER collective Thomson scattering-Preparing to diagnose fusion-born alpha particles (invited).

The ITER Collective Thomson scattering (CTS) diagnostic will measure the dynamics of fusion-born alpha particles in the burning ITER plasma by scattering a 1 MW 60 GHz gyrotron beam off fast-ion induced fluctuations in the plasma. The diagnostic will have seven measurement volumes across the ITER cross section and will resolve the alpha particle energies in the range from 300 keV to 3.5 MeV; importantly, the CTS diagnostic is the only diagnostic capable of measuring confined alpha particles for energies below ∼1.7 MeV and will also be sensitive to the other fast-ion populations. The temporal resolution is 100 ms, allowing the capture of dynamics on that timescale, and the typical spatial resolution is 10-50 cm. The development and design of the in-vessel and primary parts of the CTS diagnostic has been completed. This marks the beginning of a new phase of preparation to maximize the scientific benefit of the diagnostic, e.g., by investigating the capability to contribute to the determination of the fuel-ion ratio and the bulk ion temperature as well as integrating data analysis with other fast-ion and bulk-ion diagnostics.

Spin-coated freestanding films for biomedical applications.

Spin-coating is a widely employed technique for the fabrication of thin-film coatings over large areas with smooth and homogeneous surfaces. In recent years, research has extended the scope of spin-coating by developing methods involving the interface of the substrate and the deposited solution to obtain self-supported films, also called freestanding films. Thereby, such structures have been developed for a wide range of areas. Biomedical applications of spin-coated freestanding films include wound dressings, drug delivery, and biosensing. This review will discuss the fundamental physical and chemical processes governing the conventional spin-coating as well as the techniques to obtain freestanding films. Furthermore, developments within this field with a primary focus on tissue engineering applications will be reviewed.

Genetic control of the B cell response to LPS: opposing effects in peritoneal versus splenic B cell populations.

Lipopolysaccharide (LPS) from gram-negative bacteria activates B cells, enabling them to proliferate and differentiate into plasma cells. This response is critically dependent on the expression of TLR4; but other genes, such as RP105 and MHC class II, have also been shown to contribute to B cell LPS response. Here, we have evaluated the role of genetic control of the B cell response to LPS at the single cell level. We compared the response to LPS of peritoneal cavity (PEC) and splenic B cells on the BALB/c genetic background (LPS-low responder) to those on the C57BL/6J background (LPS-high responder) and their F1 progeny (CB6F1). Both PEC and splenic B cells from B6 exhibited 100% clonal growth in the presence of LPS; whereas, BALB/c PEC and splenic B cells achieved only 50% and 23% clonal growth, respectively. Adding CpG to the LPS stimulus pushed PEC B cell clonal growth in the low responder strain BALB/c up to 90%, showing that the nonresponse to LPS is a specific effect. Surprisingly, PEC B cells on the F1 background behaved as high responders, while splenic B cells behaved as low responders to LPS. The data presented here reveals a previous unsuspected behavior in the genetic control of the B cell response to LPS with an opposing impact in splenic versus peritoneal cavity B cells. These results suggest the existence of an, as yet, unidentified genetic factor exclusively expressed by coelomic B cells that contributes to the control of the LPS signaling pathway in the B lymphocyte.

Lymphogranuloma venereum serovar L2b in Portugal.

Chlamydia trachomatis, serovar L2, is the causative agent of lymphogranuloma venereum (LGV), which during recent years has been responsible for various outbreaks reported among men who have sex with men (MSM) in Western Europe, America, Canada and Australia. Samples from nine patients with chronic proctitis, seen at a local hospital were sent to us for identification of C. trachomatis serovar L2. The presence of C. trachomatis serovar L DNA was identified by realtime polymerase chain reaction (PCR) in two patients. They both had high positive C. trachomatis antibody titres (>/=10,000) and were found to be infected with serovar L2b by sequencing after amplification of the omp 1 gene by a nested PCR technique. These two individuals met the diagnostic criteria for LGV serovar L2b infection and, to our knowledge, these are the first cases described in Portugal.

Optimization of silver-containing bioglass nanoparticles envisaging biomedical applications.

Bioglass nanoparticles (BGs) are of outmost importance in the biomedical field, because their unique characteristics, namely osteoconductivity and osteoinductivity, and also in certain conditions, angiogenic and bactericidal properties. In this work, novel bioglass nanoparticles containing silver (AgBGs) were synthesized by a sol-gel method, adopting different thermal treatments to obtain new nanoparticles with bioactive and antibacterial features. This is the first systematic study of the effect of the thermal treatment on the properties of AgBGs. The effect of the studied thermal treatments on the properties of synthesized nanoparticles was analyzed by several characterization techniques: FT-IR, XRD, S-TEM, SEM-EDS and Zeta potential. FT-IR allowed the identification of the characteristic peaks of the nanoparticles and XRD revealed the presence of the characteristic peaks of an apatite-like phase. By S-TEM analysis it was found that the produced nanoparticles are dense and have a diameter <200 nm. The SEM micrographs showed their surface morphology and Zeta potential measurements were performed to study their suspension stability. Additionally, in vitro bioactivity tests confirmed their bioactive potential and the microbiological tests evidenced their bactericidal effect. These promising AgBGs could be incorporated either in 2D or 3D structures for several biomedical applications, namely in the orthopedic and dental fields.

Antibacterial free-standing polysaccharide composite films inspired by the sea.

The adhesive capabilities of marine mussel proteins are well-known, exhibiting the ability to stick to different underwater substrates, either inorganic or organic. These unique adhesive properties are due to the high levels of amino acid, 3,4-dihydroxyphenyl-l-alanine (DOPA), which presents the reactive catechol group. Herein, novel antibacterial free-standing (FS) films were developed with natural polymers, namely chitosan (CHT) and hyaluronic acid (HA), being the catechol-functionalized hyaluronic acid (HA-DN) also included to provide wet adhesive properties. In order to obtain composite films, silver doped bioglass nanoparticles (Ag-BGs) were incorporated to promote bactericidal and bioactive properties, being tested four distinct formulations of FS films. Their surface morphology and topography, wettability, weight loss, swelling, mechanical, adhesion and bioactivity was analyzed. In particular, bioactivity tests revealed that upon immersion in simulated body fluid, there was the formation of a bone-like apatite layer. Moreover, upon 16 h in direct contact with Staphylococcus aureus and Escherichia coli cultures, these FS films exhibited a clear antibacterial effect. Therefore, such bioactive, antibacterial and adhesive free-standing films could potentially be used as temporary guided bone regeneration films, in particular to regenerate small bone defects and also periodontal tissues.

Antioxidant effect of the pequi oil (Caryocar brasiliense) on the hepatic tissue of rats trained by exhaustive swimming exercises.

Increased oxygen consumption and activation of specific metabolic pathways during or after physical exercise lead to the formation of reactive oxygen and nitrogen species. An investigation was made into the effects of pequi oil supplementation in protecting liver cells against injury resulting from oxidative stress. The experiments involved 20 male adult Wistar rats ( Rattus norvegicus). The animals were divided into four experimental groups: Group 1: sedentary control group; Group 2: exercise control group; Group 3: supplemented sedentary group; and Group 4: supplemented exercise group. Supplementation consisted of pequi oil administered by oral gavage (400 mg). The animals of the exercised groups were subjected to 20 swimming sessions for 5 weeks (with progressive increase of 10 minutes until exhaustion). Samples were collected from the right hepatic lobe for histopathological analysis and determination of malondialdehyde levels. The histopathological analyses revealed that the animals of the exercised control group had moderate liver damage, while the animals of the supplemented exercised group had slight tissue damage, and the sedentary control and sedentary supplemented groups showed no tissue damage. The malondialdehyde levels showed higher and statistically significant in exercise control group when compared to the other evaluated groups (p<0.05). In conclusion the supplementation with pequi oil had a protective effect on liver cells against damage caused by oxygen free radicals during strenuous exercise, as demonstrated by the indicator of lipid peroxidation.

Anorectal lymphogranuloma venereum: the first two confirmed cases in Portugal.

We describe two cases of lymphogranuloma venereum (LGV) in men who have sex with men in Portugal in 2008. These first two confirmed cases of LGV L2b proctitis in Portugal highlight the need for an enhanced surveillance programme in Portugal.

Effectiveness of furosemide in attenuating exercise-induced pulmonary haemorrhage in horses when administered at 4- and 24-h prior to high-speed training.

BACKGROUND: Due to the high prevalence of EIPH in racehorses and its potential impact on the horse's health, furosemide administration is permitted up to 4-h prior to post time in most North American racing jurisdictions. Anecdotal reports suggest that administration of furosemide 24-h prior to strenuous exercise may be equally effective in decreasing the severity of EIPH. OBJECTIVES: To 1) compare the efficacy of furosemide in reducing the presence and severity of EIPH when administered 4- or 24-h prior to strenuous exercise 2) characterise electrolyte and blood parameters following administration of furosemide at 4- and 24-h prior to exercise. STUDY DESIGN: 3-way crossover. METHODS: Fifteen Thoroughbred racehorses received 5 mL of 0.9% NaCl or 250 mg of furosemide either 4- or 24-h prior to a 5-furlong simulated race. Blood samples were collected prior to and post-run for determination of furosemide, lactate, haemoglobin and electrolyte concentrations. One-hour post-race, an endoscopic exam and bronchoalveolar lavage (BAL) were performed. Horses were assigned an EIPH score based on predetermined criteria and the number of red blood cells in BAL fluid was determined. RESULTS: Endoscopic EIPH scores were lower in the 4-h vs. the 24-h (P = 0.03) furosemide groups. RBC counts in BAL fluid were lower in the 4-h furosemide vs. saline treatment groups (P = 0.01) but no difference was noted between the saline and 24-h furosemide groups (P = 0.3), nor between the 4- and 24-h groups (P = 0.5). MAIN LIMITATIONS: Small sample size and large range of running times for the 5-furlong work. CONCLUSIONS: While none of the treatments prevented EIPH, endoscopic scores and RBC counts in BAL fluid support the efficacy of furosemide in reducing the severity of EIPH. Endoscopic scores were lower in the 4-h furosemide group compared with 24-h administration. Red blood cell counts were lower in the 4-h furosemide group compared with saline treatment.

Overdose in young children treated with anti-reflux medications: Poisons enquiry evidence of excess 10-fold dosing errors with ranitidine.

BACKGROUND: Accidental drug overdose is a common problem in young children. We examined the influence of formulation and dose in enquiries for different gastro-oesophageal reflux disease treatments in children under 5 years to the UK's National Poisons Information Service. METHODS: Overdose characteristics with ranitidine, omeprazole or domperidone were compared with those of metoclopramide and the H-1 antagonist chlorphenamine, for the period 1 July 2007 to 30 June 2015. RESULTS: There were a total of 1092 ranitidine, 618 domperidone and 1193 omeprazole cases; 669, 281 and 424, respectively, were single agent enquiries; of these 77% (517) of ranitidine, 52% (145) domperidone and 32% (135) omeprazole cases occurred in children <5 years. In comparison, 17% (34/424) of metoclopramide and 53% (533/1013) of chlorphenamine were <5 years; 79% (410/517) of ranitidine overdose enquiries in children <5 years were under 6 months of age, higher than domperidone (68/145, 47%; p < 0.05), omeprazole (8/135, 6%), chlorphenamine (13/553, 2%) or metoclopramide (1/34, 3%) (all p < 0.01). In children aged <6 months, 101 were 10-fold overdoses, 86 with ranitidine. CONCLUSIONS: Tenfold overdoses in children (<5 years) were a feature of ranitidine enquiries, likely due to the high concentration of the syrup. This has relevance to other liquid formulations used for non-licenced indications in young children. Such therapeutic errors cause significant carer anxiety and healthcare utilization. Assistance is needed from manufacturers and legislators in modifying formulation so that drugs can be safely used in young children. Education of prescribers and carers is also needed to reduce the incidence of such errors that cause significant carer anxiety and healthcare utilization.

Organizing pneumonia revisited: insights and uncertainties from a series of 67 patients.

Background: Organizing pneumonia (OP) is classified as an acute/subacute pneumonia according to the American Thoracic Society/European Respiratory Society statement (2013 update). Although its clinical presentation, radiologic and histologic features are well established, data on the relevance of potential causes, corticosteroid doses and length, or management of relapses are based on heterogeneous series of patients. Objectives: The aims of this study were to describe clinical presentation, diagnosis and treatment of OP, explore potential causes, discuss strategies for managing relapses, and analyze prognostic factors. We also discuss our findings in relation to relevant data in the literature. Methods: We performed a cross-sectional study of all patients diagnosed with OP at a tertiary referral center in northern Portugal between 2008 and 2015. Results: Sixty-seven patients were diagnosed with OP over the 7-year study period. Dyspnea and cough were the most common presenting symptoms and approximately 30% of patients were hospitalized at the time of diagnosis. Approximately half of the patients were receiving drugs described as potential causes of OP. Microorganisms were isolated in approximately one-third of patients. Other potential causes identified were hematologic disorders, neoplasms, connective tissue diseases, myelodysplastic syndromes, immunodeficiencies, radiotherapy, and bird exposure. Cryptogenic OP was diagnosed in just 16 patients (23.8%). Corticosteroids were the most common treatment and 11 patients (16.4%) experienced relapse. Conclusions: The findings for this series of patients confirm the extreme variability of the contexts in which OP can occur and suggest that rather than a distinct, homogeneous clinicopathologic entity, OP is a non-specific reaction whose outcomes are dependent on the cause. (Sarcoidosis Vasc Diffuse Lung Dis 2018; 35: 129-138).

Riboflavin did not provide anti-inflammatory or antioxidant effects in an experimental model of sepsis

We aimed to evaluate whether the administration of riboflavin to septic animals reduces inflammation, oxidative stress, organ dysfunction, and mortality. C57BL/6 mice, 6-8 weeks old, were allocated to the study group (polymicrobial sepsis induced by cecal ligation and puncture (CLP) + antibiotic + iv riboflavin), control (CLP + antibiotic + iv saline), or naïve (non-operated controls). Serum concentrations of alanine aminotransferase (ALT), creatine kinase-MB (CK-MB), urea, and creatinine, and markers of inflammation [interleukin (IL)-6, tumor necrosis factor (TNF)-α, keratinocyte-derived chemokine (KC), and macrophage inflammatory protein (MIP)-2)], and oxidative stress (malondialdehyde (MDA) were measured 12 h after the experiment. Animal survival rates were calculated after 7 days. Means between groups were compared using linear regression models adjusted under the Bayesian approach. No significant difference was observed between control and study groups in serum concentrations of IL-6 (95% credible interval) (-0.35 to 0.44), TNF-α (-15.7 to 99.1), KC (-0.13 to 0.05), MIP-2 (-0.84 to 0.06), MDA (-1.25 to 2.53), or ALT (-6.6 to 11.5). Serum concentrations of CK-MB (-145.1 to -30.1), urea (-114.7 to -15.1), and creatinine (-1.14 to -0.01) were higher in the study group. Survival was similar in both groups (P=0.8). Therefore, the use of riboflavin in mice undergoing sepsis induced by CLP did not reduce inflammation, oxidative stress, organ dysfunction, or mortality compared with placebo.