The evolution of antimicrobial peptide resistance in Pseudomonas aeruginosa is severely constrained by random peptide mixtures.

The prevalence of antibiotic-resistant pathogens has become a major threat to public health, requiring swift initiatives for discovering new strategies to control bacterial infections. Hence, antibiotic stewardship and rapid diagnostics, but also the development, and prudent use, of novel effective antimicrobial agents are paramount. Ideally, these agents should be less likely to select for resistance in pathogens than currently available conventional antimicrobials. The usage of antimicrobial peptides (AMPs), key components of the innate immune response, and combination therapies, have been proposed as strategies to diminish the emergence of resistance. Herein, we investigated whether newly developed random antimicrobial peptide mixtures (RPMs) can significantly reduce the risk of resistance evolution in vitro to that of single sequence AMPs, using the ESKAPE pathogen Pseudomonas aeruginosa (P. aeruginosa) as a model gram-negative bacterium. Infections of this pathogen are difficult to treat due the inherent resistance to many drug classes, enhanced by the capacity to form biofilms. P. aeruginosa was experimentally evolved in the presence of AMPs or RPMs, subsequentially assessing the extent of resistance evolution and cross-resistance/collateral sensitivity between treatments. Furthermore, the fitness costs of resistance on bacterial growth were studied and whole-genome sequencing used to investigate which mutations could be candidates for causing resistant phenotypes. Lastly, changes in the pharmacodynamics of the evolved bacterial strains were examined. Our findings suggest that using RPMs bears a much lower risk of resistance evolution compared to AMPs and mostly prevents cross-resistance development to other treatments, while maintaining (or even improving) drug sensitivity. This strengthens the case for using random cocktails of AMPs in favour of single AMPs, against which resistance evolved in vitro, providing an alternative to classic antibiotics worth pursuing.

Lizard host abundances and climatic factors explain phylogenetic diversity and prevalence of blood parasites on an oceanic island.

Host abundance might favour the maintenance of a high phylogenetic diversity of some parasites via rapid transmission rates. Blood parasites of insular lizards represent a good model to test this hypothesis because these parasites can be particularly prevalent in islands and host lizards highly abundant. We applied deep amplicon sequencing and analysed environmental predictors of blood parasite prevalence and phylogenetic diversity in the endemic lizard Gallotia galloti across 24 localities on Tenerife, an island in the Canary archipelago that has experienced increasing warming and drought in recent years. Parasite prevalence assessed by microscopy was over 94%, and a higher proportion of infected lizards was found in warmer and drier locations. A total of 33 different 18s rRNA parasite haplotypes were identified, and the phylogenetic analyses indicated that they belong to two genera of Adeleorina (Apicomplexa: Coccidia), with Karyolysus as the dominant genus. The most important predictor of between-locality variation in parasite phylogenetic diversity was the abundance of lizard hosts. We conclude that a combination of climatic and host demographic factors associated with an insular syndrome may be favouring a rapid transmission of blood parasites among lizards on Tenerife, which may favour the maintenance of a high phylogenetic diversity of parasites.

Diagnostic delay in patients with sporadic hereditary transthyretin-mediated amyloidosis.

BACKGROUND: Hereditary transthyretin-mediated amyloidosis (ATTRv amyloidosis) is a rare progressively incapacitating condition with a wide range of genotype/phenotype presentations. It is frequently diagnosed late in its course, particularly in sporadic cases. OBJECTIVES: Analysing predictors of diagnostic delay in this subpopulation should be, therefore, a priority. METHODS: 109 apparently sporadic ATTRv amyloidosis patients followed in a reference centre in Hospital de Santa Maria (ULS Santa Maria-CAML), in Lisbon, were studied. Time from symptom onset to diagnosis, age, sex, municipality of origin and initial symptoms were obtained. Diagnostic delay was compared between different decades with a Kruskal-Wallis test, and its predictors were evaluated in a univariate model followed by a binary logistic regression analysis to calculate the adjusted odds ratio. RESULTS: The median diagnostic delay was 1262 days. There was a non-significant difference in diagnostic delay between the 80 s, 90 s, 2000s and 2010s decades. There was a non-significant trend for a longer diagnostic delay in woman and in patients having no neurologic symptoms at onset. CONCLUSION: There is an important diagnostic delay in sporadic cases of ATTRv amyloidosis. Awareness should be spread among clinicians regarding the various manifestations of this disease, stressing the importance of family history and epidemiological data.

Landscape genetics reveals contrasting patterns of connectivity in two newt species (Lissotriton montandoni and L. vulgaris).

Ecologically distinct species may respond to landscape changes in different ways. In addition to basic ecological data, the extent of the geographic range has been successfully used as an indicator of species sensitivity to anthropogenic landscapes, with widespread species usually found to be less sensitive compared to range-restricted species. In this study, we investigate connectivity patterns of two closely related but ecologically distinct newt species - the range-restricted, Lissotriton montandoni and the widespread, L. vulgaris - using genomic data, a highly replicated setting (six geographic regions per species), and tools from landscape genetics. Our results show the importance of forest for connectivity in both species, but at the same time suggest differential use of forested habitat, with L. montandoni and L. vulgaris showing the highest connectivity at forest-core and forest-edges, respectively. Anthropogenic landscapes (i.e., higher crop- or urban-cover) increased resistance in both species, but the effect was one to three orders of magnitude stronger in L. montandoni than in L. vulgaris. This result is consistent with a view of L. vulgaris as an ecological generalist. Even so, currently, the negative impact of anthropogenic landscapes is mainly seen in connectivity among L. vulgaris populations, which show significantly stronger isolation and lower effective sizes relative to L. montandoni. Overall, this study emphasizes how habitat destruction is compromising genetic connectivity not only in endemic, range-restricted species of conservation concern but also in widespread generalist species, despite their comparatively lower sensitivity to anthropogenic landscape changes.

Multimethod analysis of three rotary instruments produced by electric discharge machining technology.

AIM: This study aimed to compare three rotary instruments produced by the EDM process with the heat-treated ProTaper Gold system regarding design, metallurgy, mechanical properties and shaping ability. METHODOLOGY: HyFlex EDM (25/~), Neoniti (25/.06), EDMax (25/.06) and ProTaper Gold (25/.08v) instruments (n = 58 per group) were compared regarding design, metallurgy and mechanical performance. Unprepared canal areas were calculated for each system after preparation of mesiobuccal, mesiolingual and distal canals of mandibular molars (15 canals per group) using micro-CT technology. Statistical analyses were performed using One-way anova post-hoc Tukey and Kruskal-Wallis post-hoc Dunn's tests (α = 5%). RESULTS: All instruments had asymmetrical blades, no radial lands, no major defects and almost equiatomic nickel/titanium ratios, but different cross-section designs, tip geometries and surface appearances. Although instruments had distinct transformation temperature curves, they showed crystallographic martensitic arrangement at 21°C and mixed austenite plus R-phase at body temperature. Neoniti and HyFlex EDM showed similar results in all mechanical tests (p > .05), while EDMax and ProTaper Gold had similar time to fracture (p = .841), maximum bending load (p = .729), and cutting ability (p = .985). ProTaper Gold showed the highest torque to failure (p < .001) and HyFlex EDM had the lowest buckling resistance (p < .001). Mean percentages of unprepared canal areas ranged from 20.4% to 25.7% in the mesial canals, and from 20.8% to 26.2% in the distal canal, with no statistical differences among systems (p > .05). CONCLUSIONS: Instruments' geometry and phase transformation temperatures influenced the results of the mechanical tests, but not their shaping ability.

Excretory urographic, ultrasonographic, and cystoscopic characteristics of urinary bladder abscesses in a dog.

A 7-year-old male neutered, 6.5 kg, Maltese breed dog was referred with a history of dysuria after undergoing two cystotomies to remove calcium oxalate uroliths. Survey radiographs were inconclusive. Excretory urography allowed visualization of two filling defects in the cranioventral urinary bladder region. Ultrasonography showed urinary bladder sediments, a diffusely thickened wall, and two circumscribed intramural/intraluminal structures with an anechoic interior and a smooth and regular surface. The cystoscopic diagnosis was urinary bladder abscesses and urine cultures were positive for Escherichia coli. Antimicrobial/anti-inflammatory therapy was started with good clinical outcomes and negative culture after 21 days.

The Secretome of Human Neonatal Mesenchymal Stem Cells Modulates Doxorubicin-Induced Cytotoxicity: Impact in Non-Tumor Cells.

Doxorubicin (Dox) is one of the most widely used treatments for breast cancer, although limited by the well-documented cardiotoxicity and other off-target effects. Mesenchymal stem cell (MSC) secretome has shown immunomodulatory and regenerative properties, further potentiated under 3D conditions. This work aimed to uncover the effect of the MSC-derived secretome from 3D (CM3D) or 2D (CM2D) cultures, in human malignant breast cells (MDA-MB-231), non-tumor breast epithelial cells (MCF10A) and differentiated AC16 cardiomyocytes, co-treated with Dox. A comprehensive proteomic analysis of CM3D/CM2D was also performed to unravel the underlying mechanism. CM3D/CM2D co-incubation with Dox revealed no significant differences in MDA-MB-231 viability when compared to Dox alone, whereas MCF10A and AC16 viability was consistently improved in Dox+CM3D-treated cells. Moreover, neither CM2D nor CM3D affected Dox anti-migratory and anti-invasive effects in MDA-MB-231. Notably, Ge-LC-MS/MS proteomic analysis revealed that CM3D displayed protective features that might be linked to the regulation of cell proliferation (CAPN1, CST1, LAMC2, RANBP3), migration (CCN3, MMP8, PDCD5), invasion (TIMP1/2), oxidative stress (COX6B1, AIFM1, CD9, GSR) and inflammation (CCN3, ANXA5, CDH13, GDF15). Overall, CM3D decreased Dox-induced cytotoxicity in non-tumor cells, without compromising Dox chemotherapeutic profile in malignant cells, suggesting its potential use as a chemotherapy adjuvant to reduce off-target side effects.

Pyridine-Containing Macrocycles Display MMP-2/9 Inhibitory Activity and Distinct Effects on Migration and Invasion of 2D and 3D Breast Cancer Models.

The role of metalloproteinases (MMPs) on the migration and invasion of cancer cells has been correlated with tumor aggressiveness, namely with the up-regulation of MMP-2 and 9. Herein, two pyridine-containing macrocyclic compounds, [15]pyN5 and [16]pyN5, were synthesized, chemically characterized and evaluated as potential MMP inhibitors for breast cancer therapy using 3D and 2D cellular models. [15]pyN5 and [16]pyN5 (5-20 µM) showed a marked inhibition of MMPs activity (100% at concentrations ≥ 7.5 µM) when compared to ARP-100, a known MMP inhibitor. The inhibitory activity of [15]pyN5 and [16]pyN5 was further supported through in silico docking studies using Goldscore and ChemPLP scoring functions. Moreover, although no significant differences were observed in the invasion studies in the presence of all MMPs inhibitors, cell migration was significantly inhibited by both pyridine-containing macrocycles at concentrations above 5 µM in 2D cells (p < 0.05). In spheroids, the same effect was observed, but only with [16]pyN5 at 20 µM and ARP-100 at 40 µM. Overall, [15]pyN5 and [16]pyN5 led to impaired breast cancer cell migration and revealed to be potential inhibitors of MMPs 2 and 9.

In the Brain, It Is Not All about Sugar.

The maintenance of energetic homeostasis relies on a tight balance between glycolysis and mitochondrial oxidative phosphorylation. The case of the brain is a peculiar one, as although entailing a constant demand for energy, it is believed to rely mostly on glucose, particularly at the level of neurons. Nonetheless, this has been challenged by studies that show that alternatives such as lactate, ketone bodies, and glutamate can be used as fuels to sustain neuronal activity. The importance of fatty acid (FA) metabolism to this extent is still unclear, albeit sustaining a significant energetic output when compared to glucose. While several authors postulate a possible role of FA for the energetic homeostasis of the brain, several others point out the intrinsic features of this pathway that make its contribution difficult to explain in the context of neuronal bioenergetics. Moreover, fueling preference at the synapse level is yet to be uncovered. In this review, we discuss in detail the arguments for and against the brain usage of FA. Furthermore, we postulate that the importance of this fuel may be greater at the synapse, where local mitochondria possess a set of features that enable a more effective usage of this fuel source.

Effects of Hyperbaric Oxygen Therapy on Hemogram, Serum Biochemistry and Coagulation Parameters of Dogs Undergoing Elective Laparoscopic-Assisted Ovariohysterectomy.

BACKGROUND: This study explored the effects of hyperbaric oxygen therapy (HBOT) on hemogram, serum biochemistry and hemostatic variables in female dogs undergoing laparoscopic-assisted ovariohysterectomy (OVH). MATERIALS: Thirty adult, mixed-breed, healthy female dogs were randomly divided into the following three groups: HBOT + SURG (exposed to two absolute atmospheres (ATAs) for 45 min followed by laparoscopic-assisted OVH), HBOT (exposed to two ATAs for 45 min) and SURG (laparoscopic-assisted OVH). Blood samples were collected at T0 (at the admission), at T1, 24 h after T0 (immediately after HBOT in the HBOT + SURG and HBOT groups, and immediately before anesthetic premedication in the SURG group), and at T2, 48 h after T0 (24 h after HBOT and anesthetic premedication). METHODS: Assessments included erythrogram, leukogram, thrombogram, renal and hepatic serum biochemistry, prothrombin time (PT), activated partial thromboplastin time (APTT), buccal mucosal bleeding time (BMBT) and bloodstain area (BA) on hygroscopic paper collected at the BMBT. RESULTS: Both the HBOT + SURG and SURG groups presented neutrophilia (p ≤ 0.0039) at T2 and an increase of ALP at T2 (p ≤ 0.0493), the SURG group presented an increase in leukocyte count at T2 (p = 0.0238) and the HBOT + SURG group presented a reduction in lymphocyte count at T2 (p = 0.0115). In the HBOT + SURG group, there was a reduction in PT and APTT in relation to the baseline value (p ≤ 0.0412). CONCLUSIONS: A session of HBOT at two ATAs for 45 min did not cause changes in the BMBT or BA in healthy female dogs. Some blood parameters investigated (neutrophil and lymphocyte count, ALP, PT and APTT) were affected by the use of HBOT.

Nephrectomy in 52 dogs for the treatment of Dioctophyme renale infection - Knowing the enemy to win.

Dioctophyme renale (D. renale) is a nematode that parasitizes the kidney of mammals. Treatment is often surgical, with removal of the affected organ. This retrospective study aims to evaluate the epidemiological, clinical, and surgical aspects, the interval between diagnosis and treatment, the occurrence of pre- and intraoperative complications, and the postoperative survival time of dogs parasitized by D. renale undergoing therapeutic nephrectomy. Records of fifty-two dogs treated in a single hospital service were analyzed. We collected epidemiological data, laboratory results, diagnostic method, anesthetic protocol, surgical technique and time, type of antimicrobial prophylaxis, pre- and intraoperative complications, location and number of parasites, and postoperative survival time. Of the 52 dogs undergoing right nephrectomy by laparotomy, 61.5 % were female and 63.4 % were adults. Although the most common clinical sign was hematuria (25 %), 61.5 % of the patients were asymptomatic. Eosinophilia and increased serum urea were the only laboratory changes found. The interval between diagnosis and surgery was 27.4 ± 23 days and no patient showed changes suggestive of surgical emergency. The most common surgical approach was the right paracostal (61.5 %), and a continuous suture pattern was predominant. Intraoperative complications occurred in 9.6 % of the procedures, varying from mild to severe hemorrhage. Mean postoperative survival was 835.5 ± 428 days. Dioctophymosis was effectively controlled by nephrectomy of the affected kidney, allowing a mean survival of more than 830 days. No serious complications caused by intervals between diagnosis and treatment have been reported. This is the largest retrospective study evaluating dogs infected with D. renale that were surgically treated.

Functionalization of polydimethylsiloxane membranes to be used in the production of voice prostheses.

The voice is produced by the vibration of vocal cords which are located in the larynx. Therefore, one of the major consequences for patients subjected to laryngectomy is losing their voice. In these cases, a synthetic one-way valve set (voice prosthesis) can be implanted in order to allow restoration of speech. Most voice prostheses are produced with silicone-based materials such as polydimethylsiloxane (PDMS). This material has excellent properties, such as optical transparency, chemical and biological inertness, non-toxicity, permeability to gases and excellent mechanical resistance that are fundamental for its application in the biomedical field. However, PDMS is very hydrophobic and this property causes protein adsorption which is followed by microbial adhesion and biofilm formation. To overcome these problems, surface modification of materials has been proposed in this study. A commercial silicone elastomer, SylgardTM 184 was used to prepare membranes whose surface was modified by grafting 2-hydroxyethylmethacrylate and methacrylic acid by low-pressure plasma treatment. The hydrophilicity, hydrophobic recovery and surface energy of the produced materials were determined. Furthermore, the cytotoxicity and antibacterial activity of the materials were also assessed. The results obtained revealed that the PDMS surface modification performed did not affect the material's biocompatibility, but decreased their hydrophobic character and bacterial adhesion and growth on its surface.

Effect of the progressive taper enlargement of buccal root canals of three-rooted maxillary molars: A stepwise micro-CT study.

This study evaluated unprepared canal areas, volume of dentine removal, and dentine thickness after the progressive enlargement of the buccal canals of 22 maxillary molars, with and without the MB2, using instruments with the same tip size (0.25 mm) but 4 different tapers (0.03, 0.05, 0.06 and 0.08 v). Micro-CT scans were performed after each instrument. Data were compared using a general linear mixed model post hoc comparisons with Kenward-Roger for Wald F tests (α = 5%). After each treatment step, it was observed an increase in the percentage volume of removed dentine and a decrease in the unprepared areas and dentine thickness at the pericervical region of all canals (p < 0.05). The percentage reduction of dentine thickness in MB2 canal was higher than in the mesiobuccal and distobuccal canals (p < 0.05). Knowledge of pre-operative dimensions and dentine thickness before enlargement of buccal canals of maxillary molars is an important factor to avoid excessive dentine removal.

Gasless laparoscopic renal biopsy in canine cadavers: a preclinical study.

In this study we aimed to analyze the feasibility of the gasless renal biopsy technique in canine cadavers. The cadavers were randomly divided into two groups: laparoscopic GCG, in which gasless laparoscopy was performed and GCP, laparoscopy with pneumoperitoneum was performed. The procedures were randomly performed on the right and left kidneys. The total surgical time, procedural steps, and intraoperative complications were recorded. The degree of difficulty of the surgical approaches was evaluated by the surgeon, assistant, and external evaluators. Renal samples were evaluated for quality, number of glomeruli, and proportion of renal cortex. The total operative time was higher in the GCG group than in the GCP group (p < 0.01). Additionally, positioning of the second portal and platform positioning took longer than the other steps. The surgical groups differed from each other in the Likert scale values for almost all the parameters regarding the difficulty of the surgical approaches (p < 0.05), with higher scores in the GCG group than in the GCP group. Based on the video recordings, the GCP group had higher scores than the GCG group for degrees of difficulty of the approach (p < 0.05). Renal histological parameters were similar between the surgical groups and surgical sides. Our study findings indicate that the proposed gasless renal biopsy technique is feasible. The longer operative time and technical difficulties in the gasless approach did not affect the quality of the renal specimens.

Neste estudo objetivamos analisar a viabilidade da técnica de biópsia renal gasless em cadáveres caninos. Os cadáveres foram divididos aleatoriamente em dois grupos: GCG laparoscópico, no qual foi realizada laparoscopia gasless e GCP, onde foi realizada laparoscopia com pneumoperitônio. Os procedimentos foram realizados aleatoriamente nos rins direito e esquerdo. O tempo cirúrgico total, etapas do procedimento e complicações intraoperatórias foram registrados. O grau de dificuldade das abordagens cirúrgicas foi avaliado pelo cirurgião, assistente e avaliadores externos. As amostras renais foram avaliadas quanto à qualidade, número de glomérulos e proporção do córtex renal. O tempo operatório total foi maior no grupo GCG do que no grupo GCP (p < 0,01). Além disso, o posicionamento do segundo portal e o posicionamento da plataforma demoraram mais do que as outras etapas. Os grupos cirúrgicos diferiram entre si nos valores da escala Likert para quase todos os parâmetros relativos à dificuldade das abordagens cirúrgicas (p < 0,05), com escores mais elevados no grupo GCG do que no grupo GCP. Com base nas gravações de vídeo, o grupo GCP obteve pontuações mais altas que o grupo GCG para graus de dificuldade da abordagem (p < 0,05). Os parâmetros histológicos renais foram semelhantes entre os grupos cirúrgicos e os lados cirúrgicos. Os resultados do nosso estudo indicam que a técnica proposta de biópsia renal gasless é viável. O maior tempo operatório e as dificuldades técnicas na abordagem sem gás não afetaram a qualidade das amostras renais.

A positive association of larval therapy and hyperbaric oxygen therapy in veterinary wound care.

The treatment of cutaneous wounds is part of the veterinary routine from initial scientific reports due to being regularly present condition. Currently, several types of treatments are available to accelerate the healing process. This report presents the case of a dog with multiple lesions in the thoracic limbs resulting from a car accident, who underwent larval therapy and hyperbaric oxygen therapy (HBOT). The animal was a 2-year-old female mixed breed dog presenting severe skin degloving, fracture in the left thoracic limb (LTL), with abrasion lesions and dislocation in the right thoracic limb (RTL). The animal underwent multiple modality therapies, such as HBOT sessions associated with larval therapy; even after the LTL presented gangrene, this treatment resulted in optimal viability of the non-necrotic tissue adjacent to the gangrene. Due to chronic pain unresponsive to drug control and the presence of a fracture at a location where a possible exoprosthesis was supposed to be fixed, the LTL ended up being amputated. There are several reports of the use of HBOT or larval therapy in traumatized limbs; however, the combination of both therapies has not been previously described in the veterinary literature. Thus, we demonstrate through this report that it was possible to quickly recover the animal with good wound resolution through tissue oxygenation and a healthy granulation bed, both provided by the therapeutic combination.

Laparoscopic nephron-sparing surgery for the treatment of canine dioctophymosis.

This paper reports on two cases of laparoscopic nephrotomy employed in the treatment of canine dioctophymosis, which is considered a unusual procedure and a new treatment proposal heretofore not performed in veterinary medicine. Two patients were treated, one with a history of hematuria and the other with incidental finding of the parasite in the abdominal cavity during elective ovariohysterectomy. Both dogs were subjected to abdominal ultrasound, which produced images indicating the presence of the parasite in the right side kidney, but with partial parenchymal preservation. The patients were therefore subjected to laparoscopic nephrotomy. The surgical procedure was effective in treating dioctophymosis and enabled minimum tissue invasion during surgery, in addition to preservation of the kidney.

Evidence of Helicobacter spp. in Saliva and Gastric Mucosa of Domestic Dogs in the Central Region of Rio Grande do Sul, Brazil.

Helicobacter pylori is a spiral-shaped bacterium, which plays a role in the aetiology of gastric diseases in humans. Non-H. pylori Helicobacter (NHPH) species naturally colonise the stomach of animals and also induce gastric lesions in humans, highlighting their zoonotic importance. We evaluated the gastric bacterial colonisation density and gastric lesions and sought to identify the main phylogenetic groups of the Helicobacter spp. obtained from dogs in the central region of Rio Grande do Sul, Brazil, with this study aiming to investigate the occurrence of Helicobacter spp. in saliva and gastric samples from these dogs. This study included 35 dogs and used analysis such as cytology, histopathology, PCR, rapid urease testing, and phylogenetic analysis. Of the dogs, 94.3% were positive for Helicobacter spp., and these bacteria were present in the stomach of 32 dogs and saliva of eight. Respectively, eight, 15, and nine dogs had mild, moderate, and severe colonisation. Lymphocytic-plasmacytic infiltrate was the main gastric lesion. However, the presence of Helicobacter and the density appeared to be unrelated to the gastric lesions. The samples possessed a high nucleotide identity with remarkably similar sequences among some of the species of NHPH such as H. heilmannii s.s., H. salomonis, H. felis, and H. bizzozeronii. The saliva of domestic dogs, even of those who appear clinically healthy, can cause Helicobacter infection in humans and other animals, with, in these dogs, increased density, occurrence rate, and predominance of NHPH of zoonotic importance being found in the stomach with a lower occurrence of Helicobacter spp. in the saliva.

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.

Evolutionary principles guiding amphibian conservation.

The Anthropocene has witnessed catastrophic amphibian declines across the globe. A multitude of new, primarily human-induced drivers of decline may lead to extinction, but can also push species onto novel evolutionary trajectories. If these are recognized by amphibian biologists, they can be engaged in conservation actions. Here, we summarize how principles stemming from evolutionary concepts have been applied for conservation purposes, and address emerging ideas at the vanguard of amphibian conservation science. In particular, we examine the consequences of increased drift and inbreeding in small populations and their implications for practical conservation. We then review studies of connectivity between populations at the landscape level, which have emphasized the limiting influence of anthropogenic structures and degraded habitat on genetic cohesion. The rapid pace of environmental changes leads to the central question of whether amphibian populations can cope either by adapting to new conditions or by shifting their ranges. We gloomily conclude that extinction seems far more likely than adaptation or range shifts for most species. That said, conservation strategies employing evolutionary principles, such as selective breeding, introduction of adaptive variants through translocations, ecosystem interventions aimed at decreasing phenotype-environment mismatch, or genetic engineering, may effectively counter amphibian decline in some areas or for some species. The spread of invasive species and infectious diseases has often had disastrous consequences, but has also provided some premier examples of rapid evolution with conservation implications. Much can be done in terms of setting aside valuable amphibian habitat that should encompass both natural and agricultural areas, as well as designing protected areas to maximize the phylogenetic and functional diversity of the amphibian community. We conclude that an explicit consideration and application of evolutionary principles, although certainly not a silver bullet, should increase effectiveness of amphibian conservation in both the short and long term.

Two-portal video-assisted colopexy in the treatment of intussusception and prolapse of the descending colon in a lamb.

A lamb presented with recurrent prolapse of the descending colon. On clinical examination, intussusception of the descending colon with the prolapse of a segment was verified. The external anal sphincter had a rupture, extending to the lacerated wound in the anus. The lamb underwent colopexy with the two-portal video-assisted incisional technique and was discharged 6 days after the surgical procedure with a satisfactory clinical outcome. There were no recurrences or complications for at least 9 months. Video-assisted colopexy is an alternative treatment for intussusception and recurrent colon prolapse in sheep, even in the presence of an external anal sphincter rupture.