Effects of Heated Pneumoperitoneum on Inflammation, Oxidative Stress, and Peritoneal Histology in Female Dogs That Underwent Video-Assisted Ovariohysterectomy.

Laparoscopic procedures require the creation of pneumoperitoneum. CO2, which must be cold and dry, is the standard gas used in such surgeries. The type of gas used, its temperature, and moisture may change the peritoneal surface and cause systemic and local oxidative stress. Our objective is to evaluate the influence of pneumoperitoneum heating on the occurrence of histological lesions in the peritoneum, inflammation, plasma oxidative stress, and on the mesothelial surface in patients undergoing video-assisted ovariohysterectomy. Twenty canine females were included and distributed evenly into two groups: heated CO2 (HG) and unheated CO2 (UHG). The biomarkers of inflammation and oxidative stress were evaluated before insufflation (T0), at 30 min (T1), and at 60 min (T2) of exposure to CO2. Biopsies of the peritoneal tissue for histological evaluation were performed at T0 and T2. Regarding plasma parameters, acetylcholinesterase (AChE) showed a greater activity in the HG at T1 (p=0.0268) and T2 (p=0.0423); in turn, butyrylcholinesterase (BChE) showed a greater activity at T2 in the HG (p=0.0175) compared with T0. Catalase activity (CAT) was different between HG times; it was higher at T1 (p=0.0253). There was a decrease in the levels of substances reactive to thiobarbituric acid (TBARS) (p=0.0117) and in glutathione (GSH) (p=0.0114) between T0 and T2 in the UHG. Regarding tissue oxidative stress, the CAT in the HG showed a greater activity at T2 than T1 (p=0.0150). By comparing the groups at each time, there was a difference only at T2 (p=0.0288), being greater in the HG. Regarding the activity of superoxide dismutase (SOD) in the HG, there was a difference between T2 in relation to T0 and T1 (p=0.0181); finally, there was an increase only at T1 (p=0.0287) in the UHG when comparing groups at the same time. There were no differences in the histological parameters evaluated. Our study demonstrates that the heating of CO2 generates a greater inflammatory response and forms reactive oxygen species (ROS) at the plasma and peritoneal levels.

Stable Redox-Cycling Nitroxide Tempol Has Antifungal and Immune-Modulatory Properties.

Invasive mycoses remain underdiagnosed and difficult to treat. Hospitalized individuals with compromised immunity increase in number and constitute the main risk group for severe fungal infections. Current antifungal therapy is hampered by slow and insensitive diagnostics and frequent toxic side effects of standard antifungal drugs. Identification of new antifungal compounds with high efficacy and low toxicity is therefore urgently required. We investigated the antifungal activity of tempol, a cell-permeable nitroxide. To narrow down possible mode of action we used RNA-seq technology and metabolomics to probe for pathways specifically disrupted in the human fungal pathogen Candida albicans due to tempol administration. We found genes upregulated which are involved in iron homeostasis, mitochondrial stress, steroid synthesis, and amino acid metabolism. In an ex vivo whole blood infection, tempol treatment reduced C. albicans colony forming units and at the same time increased the release of pro-inflammatory cytokines, such as interleukin 8 (IL-8, monocyte chemoattractant protein-1, and macrophage migration inhibitory factor). In a systemic mouse model, tempol was partially protective with a significant reduction of fungal burden in the kidneys of infected animals during infection onset. The results obtained propose tempol as a promising new antifungal compound and open new opportunities for the future development of novel therapies.

Phospholipid levels in blood during community-acquired pneumonia.

Phospholipids, major constituents of bilayer cell membranes, are present in large amounts in pulmonary surfactant and play key roles in cell signaling. Here, we aim at finding clinically useful disease markers in community-acquired pneumonia (CAP) using comprehensive phospholipid profiling in blood and modeling of changes between sampling time points. Serum samples from 33 patients hospitalized with CAP were collected at admission, three hours after the start of intravenous antibiotics, Day 1 (at 12-24 h), Day 2 (at 36-48 h), and several weeks after recovery. A profile of 75 phospholipid species including quantification of the bioactive lysophosphatidylcholines (LPCs) was determined using liquid chromatography coupled to time-of-flight mass spectrometry. To control for possible enzymatic degradation of LPCs, serum autotaxin levels were examined. Twenty-two of the 33 patients with a clinical diagnosis of CAP received a laboratory-verified CAP diagnosis by microbial culture or microbial DNA detection by qPCR. All major phospholipid species, especially the LPCs, were pronouncedly decreased in the acute stage of illness. Total and individual LPC concentrations increased shortly after the initiation of antibiotic treatment, concentrations were at their lowest 3h after the initiation, and increased after Day 1. The total LPC concentration increased by a change ratio of 1.6-1.7 between acute illness and Day 2, and by a ratio of 3.7 between acute illness and full disease resolution. Autotaxin levels were low in acute illness and showed little changes over time, contradicting a hypothesis of enzymatic degradation causing the low levels of LPCs. In this sample of patients with CAP, the results demonstrate that LPC concentration changes in serum of patients with CAP closely mirrored the early transition from acute illness to recovery after the initiation of antibiotics. LPCs should be further explored as potential disease stage biomarkers in CAP and for their potential physiological role during recovery.

Metabolomics using GC-TOF-MS followed by subsequent GC-FID and HILIC-MS/MS analysis revealed significantly altered fatty acid and phospholipid species profiles in plasma of smokers.

Mass spectrometry is an ideal tool for investigations of the metabolome in human plasma. To investigate the impact of smoking on the human metabolome, we performed an untargeted metabolic fingerprinting using GC-TOF-MS with EDTA-plasma samples from 25 smokers and 25 non-smokers. The observed elevated levels in the monounsaturated fatty acids (MUFAs) in smokers were verified by a targeted analysis using GC-FID, which revealed also significantly alterations in saturated and polyunsaturated fatty acids in smokers (p<0.05, Mann-Whitney U test). Since the main fraction of fatty acids in plasma is esterified to phospholipids, we analyzed phosphatidylcholine (PC) and phosphatidylethanolamine (PE) species composition in the plasma samples of the same subjects. The profiles of 39 PC and 40 PE species were analyzed with a newly developed and validated HILIC-ESI-MS/MS method. We were able to baseline separate the two lipid classes (PC from PE) by maintaining co-elution of individual lipid species of each class. The method shows a linear range from 0.5µM to 2000µM and an inter- and intraday coefficient of variation (CV)<20% across all analytes. Application of the validated method to the plasma samples of smokers and non-smokers, derived from a diet-controlled smoking study, revealed significantly elevated levels of PC and PE species containing MUFAs in smokers. In summary, we could demonstrate that there is a significantly altered total fatty acid profile, with increased MUFAs, in the plasma of smokers compared to non-smokers. Results obtained with the new HILIC-MS/MS method indicate that the altered fatty acid profile is also reflected in the PC and PE profile of smokers.

Epidural anesthesia and postoperatory analgesia with alpha-2 adrenergic agonists and lidocaine for ovariohysterectomy in bitches.

The aim of this study was to determine the viability and cardiorespiratory effects of the association of epidural alpha-2 adrenergic agonists and lidocaine for ovariohysterectomy (OH) in bitches. Forty-two bitches were spayed under epidural anesthesia with 2.5 mg/kg body weight (BW) of 1% lidocaine with adrenaline (CON) or in association with 0.25 mg/kg BW of xylazine (XYL), 10 µg/kg BW of romifidine (ROM), 30 µg/kg BW of detomidine (DET), 2 µg/kg BW of dexmedetomidine (DEX), or 5 µg/kg BW of clonidine (CLO). Heart rate (HR), respiratory rate (fR) and arterial pressures were monitored immediately before and every 10 min after the epidural procedure. Blood gas and pH analysis were done before, and at 30 and 60 min after the epidural procedure. Animals were submitted to isoflurane anesthesia if they presented a slightest sign of discomfort during the procedure. Time of sensory epidural block and postoperative analgesia were evaluated. All animals in CON and DEX, 5 animals in ROM and CLO, 4 animals in XYL, and 3 in DET required supplementary isoflurane. All groups, except CLO, showed a decrease in HR. There was an increase in arterial pressures in all groups. Postoperative analgesia lasted the longest in XYL. None of the protocols were totally efficient to perform the complete procedure of OH; however, xylazine provided longer postoperative analgesia than the others.

Biópsia pulmonar incisional por toracoscopia paraxifoide transdiafragmática com dois portais em cães / Pulmonary incisional biopsy by means of transdiaphragmatic paraxiphoid thoracoscopy with two ports in dog

As doenças respiratórias observadas na clínica médica de pequenos animais são numerosas, assim como as possibilidades diagnósticas, dentre as quais encontram-se os exames laboratoriais, os estudos radiográficos, os lavados broncoalveolares e as biópsias. Na presente pesquisa avaliaram-se os resultados da toracoscopia paraxifoide transdiafragmática para a realização de biópsia pulmonar em cães, sendo utilizados para tanto, 13 animais clinicamente sadios. Sob anestesia geral, produziu-se pneumotórax no volume de 30ml kg-1 de ar ambiente para cada hemitórax. O acesso foi obtido a partir de dois trocartes, posicionados entre o apêndice xifoide e o arco costal, os quais transfixaram o diafragma. O primeiro portal foi empregado para a passagem do endoscópio e o segundo para a utilização de pinça saca-bocado, empregada na obtenção de biópsia. Em seguida, a pinça foi removida e um dreno torácico foi posicionado através do portal. Durante os procedimentos, aferiram-se as frequências respiratória e cardíaca, a saturação de oxigênio, as pressões arterial média e venosa central e os parâmetros hemogasométricos. Os drenos foram removidos num período de até 48 horas de pós-operatório, verificando-se mínima produção de ar e/ou líquido. Concluiu-se se tratar de uma técnica rápida, segura e sem complicações trans e pós-operatórias, permitindo aquisição de material suficiente para a avaliação histológica do pulmão.

There are many respiratory diseases in small animals, as well as the possibilities of diagnoses, as laboratory and radiographic exams, bronchoalveolar lavage and biopsies. This research aimed at assessing the results of transdiaphragmatic paraxiphoid thoracoscopies in pulmonary biopsies of thirteen clinically healthy dogs. Under general anesthesia, each dog was insufflated with 30ml kg-1 of air in each hemithorax; two trocars were placed between the xiphoid appendix and the costal arch through the diaphragm. An endoscope was introduced by the first portal; while the second portal was used to allow access to the nippers in the biopsy forceps. Then, these nippers were removed and replaced by chest tube drainage. During the procedures, respiratory frequency, heart rate, oxygen saturation, arterial pressure and central venous pressure (CVP), hemogasometric parameters were checked. Within 48 hours after the surgery, drains were or drain was removed, with minimal production of air and/or liquid. In conclusion, this is a fast and safe technique, which is free of trans and post surgical complications, thus allowing the collection of sufficient material for pulmonary histological evaluation.