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To compare perioperative opioid consumption in dogs undergoing pelvic limb surgeries and receiving a lumbosacral plexus block by combining a lumbar [lateral pre-iliac (LPI)] block with a lumbosacral trunk [i.e. parasacral (PS group) or greater ischiatic notch plane (GIN group)] block. Study design Retrospective, descriptive, exploratory, noninferiority cohort study. Medical records of 37 client-owned dogs were analyzed. Intraoperative (primary outcome) and postoperative (24 hours) opioid use, 24 hour cumulative pain scores and prevalence of complications were compared between the two groups. Opioid use was quantified in morphine equivalents (ME, mg kg–1). The noninferiority limit for intraoperative opioid consumption was set at 0.05 ME kg–1 hour–1. Demographic data, procedure duration, surgery type and perioperative dexmedetomidine and ketamine use were also collected. A t -test or Wilcoxon rank-sum test, a Fisher's exact test and multivariable linear regression were used. Significance was set at p < 0.05. The GIN and PS groups comprised 17 and 20 dogs, respectively. Total intraoperative ME consumption was 0.17 (0.11–0.21) and 0.22 (0.16–0.30) mg kg–1 hour–1 for the GIN and PS groups, respectively (p = 0.077). The noninferiority analysis adjusted by surgery type and body mass revealed that the mean difference between the groups (GIN – PS) was –0.039 (95% CI –0.11–0.03, p = 0.247) ME mg kg–1 hour–1, indicating that the GIN group was not inferior to the PS group regarding intraoperative ME consumption. Dexmedetomidine and ketamine use, postoperative ME consumption and pain scores were similar between groups. This retrospective study and preliminary observations support the clinical use of the GIN plane block, demonstrating its noninferiority to the PS block when combined with the LPI block for multimodal perioperative analgesia in dogs undergoing pelvic limb surgeries. [ABSTRACT FROM AUTHOR]

No data in the literature have evaluated sex hormone dose escalation for treating abnormal sleep of ovariectomized rats—nor studies on the role of sex hormones in GABA synthesis of rats' sleep-related areas. The main aim of this study was to determine the maximum tolerated dose (MTD) of estradiol (ET), progesterone (PT), and the mixture of both (EPT) to restore normal sleep in a model of menopause in rats. The second purpose was to describe the in vitro activity of glutamate decarboxylase (GAD) in sleep-related brain areas in the presence or absence of sex hormones. A weekly dose-escalation design of ET, PT, or EPT was implemented in ovariectomized rats (six per group). Dose escalation continued until the dose at which 100% of the rats exhibited a state of "complete somnolence." Doses that were not toxic or did not show side effects were considered. For in vitro experiments, sleep-related brain areas were separated and incubated with radiolabeled glutamate. Estradiol (17β-E2), progesterone (P), and pyridoxal phosphate (PLP) were added to this assay, and GAD activity was determined. Under the same conditions, a second test was carried out, but the P antagonist RU486 was added to assess the role of P in GAD activity. Ovariectomy increased periodic awakenings compared to those determined for the SHAM group. The EPT for ovariectomized rats was very effective by the fifth week in decreasing arousal and achieving a similar sleep behavior to the SHAM-control group. Rats tolerated the ET, PT, and EPT well to the maximum planned dose (0.66 mg/kg and 4.4 mg/kg, respectively). No lethal events occurred; the MTD was reached. The in vitro studies indicated that the presence of 17β-E2 plus P in the assay triggered the activity of isotype 65 GAD in all the studied brain areas. RU486 in the incubation medium blocked such activity; however, the action of isotype 67 GAD was not blocked by RU486. A dose-escalation model was determined; the MTD coincided with the maximum dose of ET and PT used. However, the EPT combination restored normal sleep in the menopause model compared to the SHAMs without toxic effects. The in vitro model demonstrated that 17β-E2 plus P presence in the assay increased the activity of GAD65 in the studied brain tissues. [ABSTRACT FROM AUTHOR]

To define the morbidity and mortality rates in goats undergoing general anesthesia at a large animal teaching hospital. Retrospective, single-cohort, observational study. Records of 193 client-owned goats. Data were collected from 218 medical records on 193 goats undergoing general anesthesia between January 2017 and December 2021. Demographic data, anesthetic management, recovery period and perianesthetic complications were recorded. Perianesthetic death was defined as anesthesia-related or anesthesia-contributory death occurring within 72 hours after recovery. Records of goats that were euthanized were reviewed to ascertain the cause of euthanasia. Each explanatory variable was individually investigated by univariable penalized maximum likelihood logistic regression, followed by multivariable analysis. Statistical significance was set at p < 0.05. Perianesthetic mortality was 7.3%, but was 3.4% when considering only goats undergoing elective procedures. Multivariable analysis showed that gastrointestinal surgeries [odds ratio (OR) 19.17, standard error (SE) 12.99, 95% confidence interval (CI) 5.08–72.33; p < 0.01] and requirement for perianesthetic norepinephrine infusion (OR 10.85, SE 8.82, 95% CI 2.21–53.33; p < 0.01) were associated with increased mortality. Maintaining other variables equal, the use of perianesthetic ketamine infusion was associated with decreased mortality (OR 0.09, SE 0.09, 95% CI 0.01–0.73; p = 0.02). Anesthesia-related or anesthesia-contributory complications included hypothermia (52.4%), bradycardia (38.1%), hypotension (35.3%), hypoxemia (14.8%), regurgitation/aspiration (7.3%), azotemia/acute renal failure (4.6%), myopathies/neuropathies (4.1%) and fever of unknown origin (2.7%). In this population, gastrointestinal surgeries and the requirement for perianesthetic norepinephrine infusion were associated with increased mortality in goats undergoing general anesthesia, while ketamine infusion may have a protective effect. [ABSTRACT FROM AUTHOR]

To evaluate the feasibility of an ultrasound-guided technique targeting the medial branches of the dorsal ramus of the spinal nerves (DRSN) by injecting a dye solution at the caudal aspect of the base of lumbar mammillary processes [i.e. the retromammillary (RM) space]. Feasibility, descriptive, anatomical study. Eleven canine cadavers. Using anatomical dissections, the relationship between the mammillary processes and the branches of the DRSN of the thoracolumbar and lumbar spine was studied in two cadavers. Subsequently, ultrasound-guided RM injections were administered in nine cadavers, randomly assigned to low volume (LV; 0.01 mL kg–1) of a dye solution at multiple points from L3 to L6 on one side, and high volume (HV; 0.4 mL kg–1) at a single L4 point on the contralateral side. Gross anatomical dissections were immediately performed after the injections. The RM injections were feasible if ultrasonographic landmarks were identifiable in at least 80% of cases, and 80% of LV injections showed medial branch staining. A one-sample binomial test was used for testing feasibility. The medial branches emerged shortly after the DRSN exited the intervertebral foramen and traveled towards the caudal aspect of the base of the mammillary process, which served as the target injection point. With LV, 36 out of 36 (100%) injected medial branches were stained, meeting the criteria of feasibility (p < 0.001). The median (range) number of stained medial branches per cadaver were 4 (4–4) and 2 (1–3), with LV and HV, respectively. Although no lateral branches were stained with LV, 1 (0–2) was stained with HV. Neither ventral branch staining nor epidural spread was noted in any cadaver. Ultrasound-guided RM injections are feasible and allow for selective staining of the medial branch of the DRSN in canine cadavers at the lumbar spine. [ABSTRACT FROM AUTHOR]

To describe the sonoanatomy of the abdominal wall in live cats and to compare the distribution pattern of two versus three ultrasound-guided transversus abdominis plane (TAP) injections using clinically applicable volumes of lidocaine–dye solution in cat cadavers. Prospective anatomical study. A total of eight client-owned healthy cats and eight cat cadavers. Ultrasound anatomy of the abdominal wall, landmarks and sites for needle access were determined in live cats. Ultrasound-guided TAP injections were performed in eight thawed cat cadavers. Volumes of 0.25 or 0.16 mL kg−1 per point of a lidocaine–dye solution were injected using either two [subcostal and preiliac (SP)] or three [subcostal, retrocostal and preiliac (SRP)] injection points, respectively. Each cadaver was then dissected to determine the injectate distribution and the number of thoracolumbar nerves stained with each approach. The target nerves were defined as the ventromedial branches of the thoracic nerves 10 (T10), T11, T12, T13 and lumbar nerves 1 (L1) and L2. Sonoanatomy was consistent with anatomy upon dissection and the TAP was identified in all cadavers. A total of 16 subcostal, 16 preiliac and nine retrocostal TAP injections were performed. The overall staining success rate of the target nerves was 66.7% and 92.6% for the SP and SPR approaches, respectively (p = 0.02). The ventromedial branches of T10, T11, T12, T13, L1 and L2 were stained in 57.1%, 100.0%, 85.7%, 28.6%, 42.9% and 85.7%, and in 66.7%, 100.0%, 100.0%, 100.0%, 88.9% and 100.0% of the cases with the SP and SRP approaches, respectively. The SRP approach allowed a broader distribution around the target nerves, whereas a staining gap was observed at T13 and L1 with the SP approach. Further studies are necessary to investigate the analgesic effect of these approaches in a clinical setting. [ABSTRACT FROM AUTHOR]

To compare the perioperative use of analgesics and complication rates in dogs administered an erector spinae plane (ESP) block or a traditional opioid-based (OP) treatment as part of analgesic management during hemilaminectomy. Retrospective cohort study. Medical records of 114 client-owned dogs. General data included demographics, duration of procedure, number of laminae fenestrated, perioperative use of steroid and non-steroidal anti-inflammatory drugs. Intra- and postoperative analgesics used in 48 hours and complications rates were compared between groups. Opioid use was expressed in morphine equivalents [ME (mg kg−1)]. Continuous data were compared using the Mann–Whitney U test and incidence of events with a Fisher's exact tests. Multiple linear regression was used to evaluate association between perioperative ME consumption (dependent variable) with other independent variables. Data are presented as median (range). Differences were considered significant when p < 0.05. Group ESP comprised 42 dogs and group OP 72 dogs. No differences were observed in the general data. Intraoperative ME was 0.65 (0.20–3.74) and 0.79 (0.19–5.60) mg kg−1 in groups ESP and OP, respectively (p = 0.03). Intraoperative infusion of lidocaine was administered intravenously (IV) to 23.8% and 68% of groups ESP and OP, respectively (p < 0.0001). Intraoperative infusion of ketamine was administered IV to 21% and 40% of groups ESP and OP, respectively (p = 0.04). Regression analysis revealed the ESP block as the only independent variable affecting the perioperative ME consumption. Pharmacological intervention to treat cardiovascular complications was administered to 21.4% and 47.2% of dogs in groups ESP and OP, respectively (p = 0.008). There were no differences in postoperative complication rates. ESP block was associated with reduced perioperative opioid consumption, intraoperative adjuvant analgesic use and incidence of pharmacological interventions to treat cardiovascular complications in dogs undergoing hemilaminectomy. [ABSTRACT FROM AUTHOR]

To compare the dye distribution following either two lateral abdominal or one lateral abdominal and one subcostal ultrasound-guided transversus abdominis plane (TAP) injections of a clinically relevant volume of dye solution in dogs. Randomized cadaveric study. A total of eight canine cadavers. On one side of each cadaver, two TAP injections were performed on the lateral aspect of the abdomen (approach LL), caudal to the last rib and cranial to the iliac crest. On the contralateral hemiabdomen, one subcostal (caudal to the costal arch) and one lateral abdominal injection (between last rib and iliac crest), were performed (approach SL). Side allocation was randomly determined. A spinal needle was introduced in-plane to the transducer for each injection of methylene blue (0.25 mL kg−1). All cadavers were dissected to assess dye distribution and number of stained target nerves. All injections were performed in the TAP. The proportion of target nerve staining was 53.5% versus 80.4% with approaches LL and SL, respectively (p = 0.005). Approach LL stained the first lumbar (L1) spinal nerve in 100% of injections and ninth thoracic (T9), T10, T11, T12, T13 and L2 were stained in 0%, 0%, 37.5%, 62.5%, 87.5% and 87.5% of injections, respectively. Approach SL stained T11, L1 and L2 in 100% of injections and T9, T10, T12 and T13 were stained in 37.5%, 87.5%, 75% and 62.5% of injections, respectively. Approach SL resulted in greater staining of nerves cranial to T12 compared with approach LL. The two approaches were equivalent in staining nerves caudal to T12. Approach SL provided a broader distribution of the injected solution than approach LL, which may result in a larger blocked area in live animals undergoing celiotomy. [ABSTRACT FROM AUTHOR]

The article presents a case study of An 18-year-old American Quarter Horse mare was referred to the University of Florida Large Animal Hospital because of signs of colic for > 7 hours. The referring veterinarian treated the mare with flunixin meglumine about 6 hours before the referral examination. Abdominal ultrasonography revealed gas distention of the large colon. A complete rectal examination was prevented by the presence of the gas-distended large colon in the pelvic canal.

To evaluate whether fentanyl administered at analgesic doses in pain-free anesthetized dogs increases the serum cortisol or blood glucose concentrations and is associated with postanesthetic dysphoria. Experimental, blinded, randomized crossover. A group of six adult research dogs. Each dog was anesthetized twice and was administered fentanyl or saline with a 15-day wash-out period. Treatment allocation was randomly assigned via a closed envelope technique. Cortisol and glucose concentrations were measured on arrival in the research room (T PRE), induction of general anesthesia (T IND), extubation (T EXT) and 1 hour after extubation (T POST). Dogs were premedicated with methadone (0.1 mg kg−1) and carprofen (4 mg kg−1) intravenously (IV), anesthesia induced with propofol (4–6 mg kg−1) IV and maintained with isoflurane in oxygen. Standard anesthesia monitoring was performed throughout the experiment. Fentanyl (5 μg kg−1 over 90 seconds followed by 7.5 μg kg−1 hour −1) or an equivalent amount in mL of saline solution were administered IV for 60 minutes. T PRE behavior scores and recovery scores were evaluated using descriptive 4-point scales. Data from serum cortisol and blood glucose concentrations were analyzed with a Split Plot on time test, whereas recovery scores were analyzed using a Wilcox match-pairs signed rank test. Statistical significance was set at p < 0.05. There were no significant increases in serum cortisol and blood glucose concentrations over time, and there were no differences between treatments. None of the dogs developed postanesthetic dysphoria with either treatment. Fentanyl, administered at analgesic doses to healthy, pain-free, anesthetized dogs, was not associated with significant changes in serum cortisol or glucose concentrations in the studied population. [ABSTRACT FROM AUTHOR]

Objective Anticholinesterase drugs may produce paradoxical neuromuscular block when administered at shallow levels of neuromuscular block. The objective of this study was to evaluate the effects of neostigmine and edrophonium when administered at near-complete reversal from nondepolarizing block in anesthetized dogs. Study design Incomplete crossover, randomized, blinded experimental study. Animals A total of 12 Beagle dogs. Methods Each dog was anesthetized twice with propofol and maintained with isoflurane and dexmedetomidine. Intravenous (IV) vecuronium (0.1 mg kg-11) was administered. When the mechanographic train-of-four (TOF) ratio had spontaneously recovered to ≥0.9, either neostigmine (0.04 mg kg-11) or edrophonium (0.5 mg kg-11) was administered IV, preceeded by atropine. Changes in twitch height or TOF ratio were measured for the next 10 minutes. Recurarization was considered to be present if values decreased by ≥10%. Results Data from four dogs in each treatment were excluded from analysis, resulting in data from five dogs administered both treatments, three dogs administered neostigmine and three dogs administered edrophonium. There was no difference between groups for age, weight, T1 and T4 twitch heights or TOF ratio values, before or after anticholinesterase administration. The TOF ratio decreased by 17% and 18% in two of the eight dogs administered neostigmine, resulting from a larger increase in T1 relative to T4. No reductions in individual twitch amplitudes were recorded in those dogs. When edrophonium was used, no cases of recurarization were observed. Conclusions and clinical relevance The results support use of edrophonium for reversal of shallow neuromuscular block. The decreases in TOF ratio recorded after neostigmine does not necessarily indicate muscular weakness. Although the clinical implications are uncertain, the results suggest that, at these doses, edrophonium may be preferable to neostigmine for reversal of shallow neuromuscular block in dogs. [ABSTRACT FROM AUTHOR]

Objectives To compare the effects of regional anaesthesia and opioid administration on adrenocortical and glycaemic responses, postoperative pain and recovery quality in dogs undergoing stifle surgery. Study design Prospective, blinded clinical study. Animals Forty-five dogs anaesthetized for tibial tuberosity advancement or tibial plateau levelling osteotomy, and 15 healthy dogs undergoing noninvasive orthopaedic diagnostic procedures. Methods The baseline behaviour of each dog was classified on a descriptive scale before anaesthesia. Dogs were anaesthetized using a standard protocol. Those undergoing surgery were randomly assigned to one of three intraoperative analgesia protocols: 1) peripheral (femoral and sciatic) nerve block (bupivacaine 0.5%; 0.15 mL kg -1 in each nerve; group PNB); 2) spinal anaesthesia (isobaric bupivacaine 0.5%; 0.05 mL kg -1; group SPI); and 3) intravenous fentanyl variable rate infusion and postoperative methadone (0.2 mg kg -1; group FEN). Blood samples were collected for measurement of cortisol and glucose concentrations on arrival (TAR), at induction (TIND), at tracheal extubation (TEXT), and 1 hour post-extubation (TPEX). After extubation, a researcher unaware of the dog's group assignment evaluated pain and recovery quality using the Glasgow Composite Measure Pain Scale (Short- Form) and a descriptive scale, respectively. Results Median recovery quality was significantly worse in group FEN (p < 0.0001) compared with all other groups. Postoperative pain scores were lower in group PNB compared with groups SPI and FEN. Cortisol and glucose concentrations increased significantly from TAR at TEXT and TPEX (p < 0.0001) in group FEN, and were unchanged from TAR in the other groups. Conclusions and clinical relevance Analgesia with a peripheral nerve block or spinal anaesthesia prevented the glycaemic and cortisol responses to surgery, promoted better recovery quality, and decreased postoperative pain scores compared with FEN. In the present study, the regional anaesthesia techniques used were found to be excellent alternatives to fentanyl administration. [ABSTRACT FROM AUTHOR]

History Eleven female dogs of different breeds undergoing unilateral radical ( n = 7) or regional abdominal mastectomy ( n = 4) received an ultrasound guided transverse abdominis plane block ( TAP-block). Physical examination Subjects showed single or multiple mammary tumours. Serum biochemistry, CBC and electrocardiogram were unremarkable. Eight animals were classified as ASA physical status II and 3 as ASA III. Management Dogs were premedicated with methadone [0.1 or 0.2 mg kg−1 intravenously (IV) or intramuscularly respectively] or fentanyl (2.5 μgkg−1 IV). Anaesthesia was induced with propofol and maintained with isoflurane or sevoflurane. Unilateral ultrasound guided TAP blocks were performed in the caudal and cranial abdomen with bupivacaine 0.25% (0.3 to 0.35 mL kg−1). Intercostal nerve blocks (T4 to T11) with bupivacaine 0.25% (0.013 to 0.04 mL kg−1) completed the blocked area in dogs undergoing radical mastectomy. Follow up The median (range) of end-expired isoflurane and sevoflurane necessary to maintain anaesthesia was 1.15 (1.07-1.22) and 2.07 (2.05-2.2) vol% respectively. A single administration of fentanyl (2.5 μg kg−1, IV) was administered to control nociception (defined as an increased heart rate or mean arterial blood pressure above 20% of the pre-incisional value) in four of 11 dogs. All dogs received carprofen (2 mg kg−1 subcutaneously) at the end of surgery. Post-operative pain, assessed for 120 minutes using the short form of Glasgow Composite Pain Scale (0-24), was always lower than 3. No rescue analgesia (allowed by the protocol) was required in this time. Conclusion Transverse abdominis plane block combined with intercostal nerve blocks may be useful to produce intraoperative anti-nociception and short term post-operative analgesia in dogs undergoing unilateral mastectomy. [ABSTRACT FROM AUTHOR]

We present an experimental study of drainage in two-dimensional porous media exhibiting bimodal pore size distributions. The role of the pore size heterogeneity is investigated by measuring separately the desaturation curves of the two pore populations. The displaced wetting fluid remains trapped in small pores at low capillary numbers and is swept only above a critical capillary number proportional to the permeability of the big pores network. Based on this observation, we derive a simple criterion for phase trapping based on the balance of viscous to capillary forces. Numerical implementation of this theory in a pore network model quantitatively tits our experimental results. This combination of approaches demonstrates quantitatively the influence of geometrical heterogeneities on drainage in porous media. [ABSTRACT FROM AUTHOR]

Abstract: In this study we have evaluated the vaccine potential of a Mycobacterium tuberculosis antigen of the PPE protein family, namely PPE44 (Rv2770c). PPE44-specific immune responses could be detected in mice acutely, chronically and latently infected with M. tuberculosis. Vaccination of mice with a plasmid DNA vaccine coding for PPE44 or recombinant PPE44 protein formulated in adjuvant generated strong cellular and humoral immune responses; immunodominant T cell epitopes were identified. Most importantly, vaccination of mice with both subunit vaccines followed by an intratracheal challenge with M. tuberculosis resulted in a protective efficacy comparable to the one afforded by BCG. Taken together these results indicate that PPE44 of M. tuberculosis is a protective antigen that could be included in novel subunit TB vaccines and that warrants further analysis. [Copyright &y& Elsevier]

Summary: DNA vaccination is a potent means for inducing strong cell-mediated immune responses and protective immunity against viral, bacterial and parasite pathogens in rodents. In an attempt to increase cross-presentation through apoptosis, the DNA-encoding caspase-2 prodomain followed by wild-type or catalytically inactive mutated caspase-3 was inserted into a plasmid encoding the 32kDa mycolyl transferase (Ag85A) from Mycobacterium tuberculosis. Transient transfection showed that the mutated caspase induced slow apoptosis, normal protein expression and NF-κB activation while wild-type caspase induced rapid apoptosis, lower protein expression and no NF-κB activation. Ag85A specific antibody production was increased by co-expressing the mutated and decreased by co-expressing the wild-type caspase. Vaccination with pro-apoptotic plasmids triggered more Ag85A specific IFN-γ producing spleen cells, and more efficient IL-2 and IFN-γ producing memory cells in spleen and lungs after M. tuberculosis challenge. Compared to DNA-encoding secreted Ag85A, vaccination with DNA co-expressing wild-type caspase increased protection after infection with M. tuberculosis, while vaccination with plasmid co-expressing mutated caspase was not protective, possibly due to the stimulation of IL-6, IL-10 and IL-17A production. [Copyright &y& Elsevier]

DNA vaccines encoding the 32 000 MW mycolyl-transferase Ag85A and the 40 000 MW phosphate-binding protein PstS-3 can elicit protective immune responses against experimental infection with Mycobacterium tuberculosis in C57BL/6 mice. Here we have analysed the vaccine potential of a combination of both antigens using plasmid vectors expressing either a fusion protein of both antigens or the separate proteins driven by two independent promoters (in pBudCE4·1 vector). Comparable levels of Ag85A specific T helper 1 (Th1) type immune responses could be induced by the two combination vaccines and the single vaccine encoding the mycolyl-transferase, whereas induction of PstS-3 specific Th1-mediated responses was impaired in both combination vaccines. In contrast, magnitude of CD8+ mediated responses against the PstS-3 protein was comparable following combination or single DNA vaccination. Antigenic competition was also observed at the antibody level; PstS-3 specific levels being lower in mice vaccinated with the fusion vector and Ag85A specific levels being lower in mice vaccinated with the combination pBudCE4·1 vector (as compared to levels obtained following single plasmid immunization). Protection against M. tuberculosis was only modestly improved in mice vaccinated with the DNA combinations. It is possible that prior activation of Ag85A specific CD4+ T cells directed against this common mycobacterial antigen leads to cross-competition for major histocompatibility complex class II-restricted peptide complexes of the Pst-3 antigen. This may have implications for future combination vaccines using Ag85. [ABSTRACT FROM AUTHOR]

Abstract: Tuberculosis (TB) caused by Mycobacterium tuberculosis (Mtb), remains a major health problem. The only currently available vaccine, BCG, confers only variable protection and an improved vaccine is urgently needed. Administration of DNA vaccines encoding the secreted mycolyl-transferase Ag85A and the surface-exposed phosphate transport receptor PstS-3 elicit an immune response capable of protecting mice challenged with Mtb. In order to combine the protection against Mtb infection induced by these two DNA vaccines, we have cloned Ag85A and PstS-3 in pBudCE4.1 vector, in which antigenic expression is controlled by two independent promoters. (BALB/cxC57BL/6)F1 mice were vaccinated with this combination vaccine and immune responses were compared to those induced by vaccination with plasmids encoding the single antigens on pBudCE4.1 or pV1J.ns-tPA backbone. Antibody and Th1 type cytokine responses against Ag85A were comparable, whereas responses against PstS-3 were clearly lower in mice vaccinated with the combination plasmid, suggesting antigenic competition with the mycolyl-transferase being the dominant antigen. [Copyright &y& Elsevier]

Abstract: DNA vaccination is a potent means for inducing strong CD4+ (Th1) and particularly CD8+ mediated immune responses and protective immunity against tuberculosis infection in mice. Here we have analyzed the potential of a DNA vaccine encoding the immunodominant mycolyl-transferase Ag85A for increasing the efficacy of the current tuberculosis vaccine Mycobacterium bovis Bacille Calmette-Guérin (BCG) in three long-term survival experiments. BALB/c mice were vaccinated with BCG either following DNA priming or prior to DNA boosting. Ag85A specific CD4+ and CD8+ mediated IFN-γ responses were increased in mice primed with DNA prior to BCG, and in BCG vaccinated mice subsequently boosted with DNA. In the latter immunization protocol, antigenic stimulation also induced significant levels of IL-17. Mice were monitored for cachexia and survival following a low dose intravenous challenge with M. tuberculosis H37Rv. Priming with Ag85A but not control DNA increased significantly the protective efficacy of the BCG vaccine as indicated by reduced cachexia and prolonged survival time: 32 weeks versus 23 weeks in one experiment and 33 weeks versus 26 weeks in another experiment (MST in control, TB infected mice: 17 weeks in both experiments). On the other hand, boosting of BCG by subsequent Ag85A DNA in saline or vaxfectin™ – or recombinant 85A protein or MVA-85A for that matter – did not augment the efficacy of BCG (MST 19–21 weeks in all vaccinated groups versus 11 weeks in control, TB infected mice). Our results demonstrate that Ag85A DNA priming can increase efficacy of BCG and that boosting protocols of BCG may possibly be hampered by the induction of ThIL-17 cells. [Copyright &y& Elsevier]

Facial expressions are essential in animal communication, and facial expression-based pain scales have been developed for different species. Automated pain recognition offers a valid alternative to manual annotation with growing evidence across species. This study applied machine learning (ML) methods, using a pre-trained VGG-16 base and a Support Vector Machine classifier to automate pain recognition in caprine patients in hospital settings, evaluating different frame extraction rates and validation techniques. The study included goats of different breed, age, sex, and varying medical conditions presented to the University of Florida's Large Animal Hospital. Painful status was determined using the UNESP-Botucatu Goat Acute Pain Scale. The final dataset comprised images from 40 goats (20 painful, 20 non-painful), with 2,253 'non-painful' and 3,154 'painful' images at 1 frame per second (FPS) extraction rate and 7,630 'non-painful' and 9,071 'painful' images at 3 FPS. Images were used to train deep learning-based models with different approaches. The model input was raw images, and pain presence was the target attribute (model output). For the single train-test split and 5-fold cross-validation, the models achieved approximately 80% accuracy, while the subject-wise 10-fold cross-validation showed mean accuracies above 60%. These findings suggest ML's potential in goat pain assessment. [ABSTRACT FROM AUTHOR]

While number of studies have shown that biological sex is a risk factor in the incidence and severity of infection-induced inflammatory diseases, the underlying mechanisms are still poorly understood. In this study, we compared the innate inflammatory response in male and female mice with group B streptococcal (GBS)-induced pneumoniae. Although male and female mice displayed similar bacterial burdens, males exhibited more innate inflammatory cytokines and chemokines and a higher proportion of infiltrating monocytes/macrophages. The analysis of the distribution of macrophage subtypes M1 (pro-inflammatory) versus M2 (anti-inflammatory) yielded a higher M1/M2 ratio in infected males compared with females. Given the importance of the chromosome X-linked microRNA-223-3p (miR-223-3p) in modulating the inflammatory process and macrophage polarization, we investigated its potential contribution in sex bias of GBS-induced innate inflammatory response. Knock-down of miR-223-3p with specific antagomiR resulted in increased inflammatory response and higher M1/M2 ratio following GBS infection. Notably, compared to male mice, we detected higher amount of miR-223-3p in macrophages from females that correlated negatively with M1 phenotype. These results suggest that differential expression of miR-233-3p may impact macrophage polarization, thereby contributing to fine-tune sex differences in inflammatory response. [ABSTRACT FROM AUTHOR]

To develop an ultrasound-guided approach to intercostal nerve injection and to compare the success rate of intercostal nerve injections using blind or ultrasound-guided technique in canine cadavers. Prospective, randomized, descriptive, experimental anatomic study. A total of 14 mid-sized adult canine cadavers. Ultrasound landmarks were identified by dissection of four cadavers and used to develop an ultrasound-guided technique. The remaining 10 cadavers were randomly assigned to blind (five cadavers) or ultrasound-guided (five cadavers) injections of the third to the ninth intercostal nerves bilaterally with 0.03 mL kg–1 of colorant per injection. The target for intercostal injections was the caudal border of the respective rib, between the internal intercostal membrane and the parietal pleura. Additionally, displacement of the parietal pleura without visible intramuscular distribution was considered the end point for ultrasound-guided injections. For each cadaver, a practitioner in training performed the blocks on one hemithorax, while an experienced practitioner performed the blocks on the opposite hemithorax. Injections were considered successful if ≥1 cm of the target nerve was stained with colorant upon dissection. Success rates and length of nerve staining were analyzed with Fisher's exact and t tests, respectively. Data were considered statistically different with p < 0.05. Success rates of blind and ultrasound-guided technique were 57.1% and 91.4%, respectively (p < 0.0001). The length of intercostal nerve staining was 3.1 ± 1.2 cm and 3.6 ± 1.0 cm using blind and ultrasound-guided techniques, respectively (p = 0.02). No differences were observed between the two practitioners for blind (p = 0.33) and ultrasound-guided techniques (p = 0.67). Ultrasound guidance improves the accuracy of intercostal nerve injections when compared with blind technique, independently of the level of expertise in regional anesthesia. [ABSTRACT FROM AUTHOR]

To investigate the distribution and nerve staining of two volumes of lidocaine–dye solution after ultrasound-guided erector spinae plane (ESP) injections in canine cadavers. Experimental cadaveric study. A total of nine canine cadavers. ESP injections were performed between the longissimus thoracis muscle and the dorsolateral edge of the ninth thoracic transverse process. Two cadavers were transversally cryosectioned after unilateral ESP injections [0.6 mL kg–1; high volume (HV)]. In seven cadavers, bilateral ESP injections with HV or low volume (0.3 mL kg–1; LV) were performed. Gadodiamide was added to the injectate for two cadavers and magnetic resonance imaging (MRI) was performed pre- and post-injection. Injectate distribution and nerve staining of the branches of the spinal nerves were recorded after gross anatomical dissection. The thoracic paravertebral and epidural spaces were examined for dye solution. Cryosections, MRI and gross dissections showed that the injectate spread dorsally to the transverse processes, over the ventromedial aspect of the longissimus thoracis muscle where the medial and lateral branches of the dorsal branches of the spinal nerves are located. LV and HV stained a median (range) of 4 (2–7) and 4 (3–8) medial branches, respectively (p = 0.52). LV and HV stained 4 (2–5) and 5 (4–7) lateral branches (p = 0.26), respectively. Ventral branches were not stained, and dye was not identified in the epidural or paravertebral spaces. and clinical relevance Medial and lateral branches were consistently stained over several spinal segments. The number of nerves stained was not different with HV or LV, and the ventral branches of the spinal nerves were not stained in any cadaver. ESP block may find a clinical application to desensitize structures innervated by the medial and lateral branches of the dorsal branches of the thoracic spinal nerves. [ABSTRACT FROM AUTHOR]

To describe the gross and ultrasound anatomy of the parasacral region and an ultrasound-guided greater ischiatic notch (GIN) plane approach aimed at staining the lumbosacral trunk (LST) in canine cadavers. To evaluate if the ultrasound-guided GIN plane approach is non-inferior to the previously described ultrasound-guided parasacral approach at staining the LST. Prospective, randomized, non-inferiority experimental anatomic study. A total of 17 (23.9 ± 5.2 kg) mesocephalic canine cadavers. Anatomic and echographic landmarks, and the feasibility of performing a GIN plane technique were evaluated using two canine cadavers. The remaining 15 cadavers had each hemipelvis randomly assigned to be administered either parasacral or GIN plane injection of 0.15 mL kg–1 dye solution. The parasacral region was dissected after injections to assess the staining of LST, cranial gluteal nerve, pararectal fossa and pelvic cavity. The stained LST were removed and processed for histological evaluation of intraneural injections. A one-sided z-test for non-inferiority (non-inferiority margin –14%) was used to statistically evaluate the success of the GIN plane versus the parasacral approach. Data were considered statistically significant when p < 0.05. The GIN plane and parasacral approach stained the LST in 100% and 93.3% of the injections, respectively. The success rate difference between treatments was 6.7% [95% confidence interval, –0.6 to 19.0%; p < 0.001 for non-inferiority]. The GIN plane and parasacral injections stained the LST for 32.7 ± 16.8 mm and 43.1 ± 24.3 mm, respectively (p = 0.18). No evidence of intraneural injection was found. The ultrasound-guided GIN plane technique resulted in nerve staining that was non-inferior to the parasacral technique and may be considered an alternative to the parasacral approach to block the LST in dogs. [ABSTRACT FROM AUTHOR]

To evaluate the impact of a 30% end-inspiratory pause (EIP) on alveolar tidal volume (V Talv), airway (V Daw) and physiological (V Dphys) dead spaces in mechanically ventilated horses using volumetric capnography, and to evaluate the effect of EIP on carbon dioxide (CO 2) elimination per breath (Vco 2 br–1), PaCO 2, and the ratio of PaO 2 -to-fractional inspired oxygen (PaO 2 :F i O 2). Prospective research study. A group of eight healthy research horses undergoing laparotomy. Anesthetized horses were mechanically ventilated as follows: 6 breaths minute–1, tidal volume (V T) 13 mL kg–1, inspiratory-to-expiratory time ratio 1:2, positive end-expiratory pressure 5 cmH 2 O and EIP 0%. Vco 2 br–1 and expired tidal volume (V TE) of 10 consecutive breaths were recorded 30 minutes after induction, after adding 30% EIP and upon EIP removal to construct volumetric capnograms. A stabilization period of 15 minutes was allowed between phases. Data were analyzed using a mixed-effect linear model. Significance was set at p < 0.05. The EIP decreased V Daw from 6.6 (6.1–6.7) to 5.5 (5.3–6.1) mL kg–1 (p < 0.001) and increased V Talv from 7.7 ± 0.7 to 8.6 ± 0.6 mL kg–1 (p = 0.002) without changing the V TE. The V Dphys to V TE ratio decreased from 51.0% to 45.5% (p < 0.001) with EIP. The EIP also increased PaO 2 :F i O 2 from 393.3 ± 160.7 to 450.5 ± 182.5 mmHg (52.5 ± 21.4 to 60.0 ± 24.3 kPa; p < 0.001) and Vco 2 br–1 from 0.49 (0.45–0.50) to 0.59 (0.45–0.61) mL kg–1 (p = 0.008) without reducing PaCO 2. The EIP improved oxygenation and reduced V Daw and V Dphys , without reductions in PaCO 2. Future studies should evaluate the impact of different EIP in healthy and pathological equine populations under anesthesia. [ABSTRACT FROM AUTHOR]

Objective To determine if neuromuscular monitoring at the pelvic limb accurately reflects neuromuscular function in the larynx after administration of rocuronium in anesthetized dogs. Study design Prospective experimental study. Animals Six healthy Beagle dogs. Methods Anesthesia was maintained in dogs with isoflurane and a continuous infusion of dexmedetomidine. Rocuronium (0.6 mg kg-1) was administered intravenously to induce neuromuscular block. Train-of-four (TOF) impulses were applied to the left recurrent laryngeal nerve (RLn) and the peroneal nerve (Pn). The evoked TOF ratio (TOFR; T4:T1) was measured with electromyography (EMG) simultaneously at the larynx and at the pelvic limb. Spontaneous recoveries of T1 to 25% (T125%) and 75% (T175%) of twitch height, and to TOFR of 0.70 and 0.90 (TOFR0.90) at each EMG site were compared. Results Data from five dogs were analyzed. Times to T125% were similar at the pelvic limb and larynx when measured by EMG; time to T175% was slower at the larynx by 6 ± 4 minutes (p = 0.012). The larynx had a slower recovery to TOFR0.70 (41 ± 13 minutes) and TOFR0.90 (45 ± 13 minutes) than did the pelvic limb [29 ± 8 minutes (p = 0.011) and 33 ± 9 minutes (p = 0.003), respectively]. When the pelvic limb EMG returned to TOFR0.70 and TOFR0.90, the larynx EMG TOFR0.70 and TOFR0.90 values were 0.32 ± 0.12 (p = 0.001) and 0.38 ± 0.13 (p = 0.001), respectively. Conclusions and clinical relevance After administration of rocuronium, neuromuscular function assessed by EMG recovered approximately 36% slower at the larynx than at the pelvic limb. The results in these dogs suggest that quantitative neuromuscular monitoring instrumented at a pelvic limb may be unable to exclude residual block at the larynx in anesthetized dogs. [ABSTRACT FROM AUTHOR]

Background: The morbidity and mortality of extraparenchymal neurocysticercosis (EP-NC) remain high and effectiveness of current medical treatment is suboptimal. Various factors have been implicated in the severity of EP-NC and in the poor response to treatment, but the possible role of host immune and endocrine systems has not yet been examined thoroughly. Methodology/Principal findings: 42 participants with EP-NC before receiving standard treatment and 25 healthy controls were included in the study. Treatment response was assessed by comparing pre/post treatment parasite volumes from 3D MRI. Prior to treatment among participants with EP-NC, specific stimulation induced an increased specific proliferative response accompanied by a significant increase in IL-4, NK, NKT, Bregs and Tregs cells, whereas in healthy controls, specific stimulation induced a significant increase in IL-1β, IL-5, CCL5, IL-6, TNF-α, NK and Bregs cells. Significant differences between participants with EP-NC and healthy controls in the specific inflammatory response were observed. Participants with EP-NC prior to treatment had significantly weaker responses of proinflammatory cytokines (IL-6, TNF-α) and NK cells, and stronger IL-4 response. Anthelmintic treatment did not promote significant peripheral immunological changes at any time, although inflammation was sustained in the cerebrospinal fluid. Serum estradiol concentration significantly decreased after anthelmintic treatment among males, and cortisol correlated negatively with IL-6 and positively with IFN-γ levels. No pre-treatment immunologic or endocrinologic parameters were significantly associated with response to treatment. Conclusion/Significance: Prior to anthelmintic treatment, EP-NC was characterized by low lymphocyte reactivity accompanied by a regulatory response, which may be involved in the lack of peripheral immunological changes during and after treatment, although a central inflammatory response was present. This weak specific peripheral response could favor the chronicity of the infection and the poor response to treatment. Our findings highlight the need for new anti-inflammatory treatment focused on the central nervous system with less systemic immunosuppressive effects. Author summary: Neurocysticercosis is a central nervous system infection with the helminth Taenia solium in its larval stage. It is a marker of poverty and remains endemic in countries of Latin America, Africa, and Asia. The severity of neurocysticercosis and the prognosis of patients depend on the location of the parasites. Extraparenchymal infection involving the subarachnoid and/or ventricular space is associated with high morbidity and mortality and a suboptimal response to current anthelminthic treatment regimens. We examined peripheral immunological parameters and hormones of 42 participants with extraparenchymal neurocysticercosis before and after treatment, and their associations with subsequent treatment response. Prior to treatment, compared with healthy controls, there was low lymphocyte reactivity with a regulatory response, which may explain the lack of peripheral immunological changes during and after treatment. This weak specific peripheral response could favor the chronicity of the infection and the poor response to treatment and should be considered in research to improve patient outcomes. [ABSTRACT FROM AUTHOR]

To describe an ultrasound-guided lateral pre-iliac (LPI) and parasacral (PS) approach in feline cadavers (phase I) and compare the perioperative analgesic use and complications in cats administered LPI and PS blocks (group PNB) or epidural anesthesia (group EPI) for pelvic limb surgery (phase II). Experimental uncontrolled, anatomic and retrospective cohort study. A group of eight feline cadavers and 52 medical records. Bilateral LPI and PS approaches with 0.1 mL kg–1 of dye to stain the femoral and obturator nerves and the lumbosacral trunk, respectively, were performed on each cadaver. Nerve staining effect was evaluated upon dissections (phase I). Perioperative analgesics use, and complication rates were retrospectively compared between groups PNB and EPI (phase II). Continuous data were compared using the Mann–Whitney U test and the prevalence of events with Fisher's exact test. Differences were considered significant when p < 0.05. Dissections revealed that the LPI approach stained 94% and 75% of the femoral and obturator nerves, respectively. The PS approach stained 100% of the lumbosacral trunks. Cats enrolled in group PNB (n = 23) were administered lower doses of intraoperative opioids than those in group EPI (n = 25) (p = 0.006). Intraoperative rescue analgesia was required in 60% and 17.4% of cats enrolled in groups EPI and PNB, respectively (p = 0.003). Group PNB required more intraoperative anticholinergics than group EPI (p = 0.02). There were no differences in postoperative pain scores, analgesic use and complication rates. The ultrasound-guided LPI and PS approach stained the femoral/obturator nerves and the lumbosacral trunk, respectively, in feline cadavers. Furthermore, PNB was associated with lower intraoperative opioid use and similar postoperative pain and analgesic use compared with epidural anesthesia in a cohort of cats undergoing surgery of the pelvic limb. [ABSTRACT FROM AUTHOR]

To develop and assess the efficacy of an ultrasound (US)-guided pericapsular hip desensitization (PHD) technique in dogs. Prospective, randomized, anatomical study and a case series. A total of 30 healthy dogs, eight canine cadavers and seven dogs with hip osteoarthritis. After studying the US anatomy of the medial aspect of the coxofemoral joint and determining an acoustic window to perform an US-guided PHD in healthy dogs, the US-guided PHD was performed bilaterally in canine cadavers. A low [(LV) 0.1 mL kg–1] and high [(HV) 0.2 mL kg–1] volume of dye was injected per hip on each cadaver. The staining of the pericapsular nerves was assessed by anatomical dissection, and comparison between LV and HV was assessed using Fisher's exact test. Then, the US-guided PHD was performed using a triamcinolone–bupivacaine solution in dogs with hip osteoarthritis. Dynamic pain response was assessed before and after injection. The canine brief pain inventory (CBPI) questionnaire was used to assess treatment efficacy and duration. The US-guided PHD could be performed by inserting the needle between the iliopsoas muscle and the periosteum of the ilium. The articular branches of the femoral and obturator nerves were stained in all cadavers using both volumes. The main femoral nerve was never stained, but the main obturator nerve was stained in 37.5% and 100% of injections using LV and HV, respectively (p = 0.026). Treated animals showed decreased dynamic pain response after the injection. Compared with baseline, CBPI scores were reduced by ≥ 50% for ≥ 12 weeks in all but one dog. The US-guided PHD with both 0.1 and 0.2 mL kg–1 volumes stained the articular branches of the femoral and obturator nerves in canine cadavers and was associated with clinical improvement in dogs with hip osteoarthritis. [ABSTRACT FROM AUTHOR]

To investigate the injectate spread and nerve staining of ultrasound-guided erector spinae plane (ESP) injections at the thoracolumbar spine in canine cadavers. Prospective, randomized, descriptive, anatomic study. A total of 15 canine cadavers. The location of the medial and lateral branches of the dorsal branches of the spinal nerves (DBSN) from the tenth thoracic (T10) to the third lumbar vertebra (L3) were identified by dissection of three cadavers. ESP injections of dye (0.5 mL kg–1) were performed in seven cadavers using as landmarks the T12 transverse process (ESP Tp) on one side and the lateral aspect of the T12 mammillary process (ESP Mp) on the opposite side. Additionally, five cadavers were injected with dye (0.5 mL kg–1) bilaterally on the lateral aspect of the L2 mammillary process (ESP Mp_L2). Nerve staining effect was analyzed after gross anatomic dissections. The number of stained nerves was analyzed using the Mann–Whitney U test. Gross anatomic dissections showed that the medial and lateral branches of the DBSN change their path in relation to the epaxial muscles caudal to T11. Approaches ESP Tp and ESP Mp at T12 stained 2 (0–2) and 3 (2–4) medial (p = 0.01) and 3 (3–4) and 2 (0–2) lateral (p = 0.03) branches, respectively. Injection ESP Mp_L2 stained 3 (2–4) medial and 2 (0–3) lateral branches. Injections ESP Mp and ESP Mp_L2 produced a preferential cranial spread from the injection site. No ventral branches of the spinal nerves were stained with either technique. These results suggest that the mammillary process should be used as anatomic landmark to perform ultrasound-guided ESP blocks in the thoracolumbar spine caudal to T11 when targeting the medial branches of the DBSN. Injections should be performed one spinal segment caudal to the level intended to desensitize. [ABSTRACT FROM AUTHOR]

• Treatment with camostat did not affect the cycle threshold change in the early phase of COVID-19 disease. • Clinical improvement was similar in patients with COVID-19 treated with camostat or a placebo. • Treatment with camostat did not affect the SARS-CoV-2 neutralizing antibody response. • This randomized controlled trial showed the safe use of 300 mg camostat three times daily in patients with COVID-19. • Camostat mesylate is not effective as an antiviral drug for ambulatory patients with COVID-19. This study aimed to assess the efficacy and safety of 300 mg camostat mesylate three times daily in a fasted state to treat early phase COVID-19 in an ambulatory setting. We conducted a phase II randomized controlled trial in symptomatic (maximum 5 days) and asymptomatic patients with confirmed COVID-19 infection. Patients were randomly assigned in a 2:1 ratio to receive either camostat mesylate or a placebo. Outcomes included change in nasopharyngeal viral load, time to clinical improvement, the presence of neutralizing antibodies, and safety. Of 96 participants randomized between November 2020 and June 2021, analyses were performed on the data of 90 participants who completed treatment (N = 61 camostat mesylate, N = 29 placebo). The estimated mean change in cycle threshold between day 1 and day 5 between the camostat and placebo group was 1.183 (P = 0.511). The unadjusted hazard ratio for clinical improvement in the camostat group was 0.965 (95% confidence interval, 0.480-1.942, P = 0.921 by Cox regression). The percentage distribution of the 50% neutralizing antibody titer at day 28 visit and frequency of adverse events were similar between the two groups. Under this protocol, camostat mesylate was not found to be effective as an antiviral drug against SARS-CoV-2. Trial registration: ClinicalTrials.gov NCT04625114 ; November 12, 2020. [ABSTRACT FROM AUTHOR]

Liver X receptors (LXRs) are key regulators of macrophage function, controlling transcriptional programs involved in lipid homeostasis and inflammation. However, exactly how LXRs modulate inflammation during infection remains unknown. To explore this, we used a mouse model of Mycobacterium tuberculosis infection. Upon intratracheal infection with M. tuberculosis, LXRs and LXR target genes were induced in CD11c+ lung and alveolar cells. Furthermore, mice deficient in both LXR isoforms, LXRalpha and LXRbeta (Lxra-/-Lxrb-/- mice), were more susceptible to infection, developing higher bacterial burdens and an increase in the size and number of granulomatous lesions. Interestingly, mice solely deficient in LXRalpha, but not those lacking only LXRbeta, mirrored the susceptibility of the Lxra-/-Lxrb-/- animals. Lxra-/-Lxrb-/- mice failed to mount an effective early neutrophilic airway response to infection and showed dysregulation of both pro- and antiinflammatory factors in CD11c+ lung cells. T cell responses were strongly affected in Lxra-/-Lxrb-/- mice, showing near-complete abrogation of the infection-induced Th1 function - and even more so Th17 function - in the lungs. Treatment of WT mice with the LXR agonists TO901317 and GW3965 resulted in a 10-fold decrease of the pulmonary bacterial burden and a comparable increase of Th1/Th17 function in the lungs. The dependence of LXR signaling on the neutrophil IL-17 axis represents what we believe to be a novel function for these nuclear receptors in resistance to M. tuberculosis infection and may provide a new target for therapeutics. [ABSTRACT FROM AUTHOR]

We report a lentiviral vector harboring the human β2-microglobulin promoter, with predominant expression in immune cells and minimal proximal enhancers to improve vector safety. This lentiviral vector efficiently transduces major dendritic cell subsets in vivo. With a mycobacterial immunogen, we observed distinct functional signatures and memory phenotype in lentiviral vector- or Adenovirus type 5 (Ad5)-immunized mice, despite comparable antigen-specific CD8+ T cell magnitudes. Compared to Ad5, lentiviral vector immunization resulted in higher multifunctional and IL-2-producing CD8+ T cells. Furthermore, lentiviral vector immunization primed CD8+ T cells towards central memory phenotype, while Ad5 immunization favored effector memory phenotype. Studies using HIV antigens in outbred rats demonstrated additional clear-cut evidence for an immunogenic advantage of lentiviral vector over Ad5. Additionally, lentiviral vector provided enhance therapeutic anti-tumor protection than Ad5. In conclusion, coupling lentiviral vector with β2-microglobulin promoter represents a promising approach to produce long-lasting, high-quality cellular immunity for vaccinal purposes. Ku et al. report a lentiviral (LV) vector system in which expression of vaccine antigens is driven by β2-microglobulin (β2m) promoter. Conducting comparative studies in mice and rats, they find that LV-based vaccine outperforms the 'gold-standard' Ad5 by inducing a polyfunctional and long-lived immune response. [ABSTRACT FROM AUTHOR]

Background: The COVID-19 pandemic has imposed an enormous burden on health care systems around the world. In the past, the administration of convalescent plasma of patients having recovered from SARS and severe influenza to patients actively having the disease showed promising effects on mortality and appeared safe. Whether or not this also holds true for the novel SARS-CoV-2 virus is currently unknown. Methods: DAWn-Plasma is a multicentre nation-wide, randomized, open-label, phase II proof-of-concept clinical trial, evaluating the clinical efficacy and safety of the addition of convalescent plasma to the standard of care in patients hospitalized with COVID-19 in Belgium. Patients hospitalized with a confirmed diagnosis of COVID-19 are eligible when they are symptomatic (i.e. clinical or radiological signs) and have been diagnosed with COVID-19 in the 72 h before study inclusion through a PCR (nasal/nasopharyngeal swab or bronchoalveolar lavage) or a chest-CT scan showing features compatible with COVID-19 in the absence of an alternative diagnosis. Patients are randomized in a 2:1 ratio to either standard of care and convalescent plasma (active treatment group) or standard of care only. The active treatment group receives 2 units of 200 to 250 mL of convalescent plasma within 12 h after randomization, with a second administration of 2 units 24 to 36 h after ending the first administration. The trial aims to include 483 patients and will recruit from 25 centres across Belgium. The primary endpoint is the proportion of patients that require mechanical ventilation or have died at day 15. The main secondary endpoints are clinical status on day 15 and day 30 after randomization, as defined by the WHO Progression 10-point ordinal scale, and safety of the administration of convalescent plasma. Discussion: This trial will either provide support or discourage the use of convalescent plasma as an early intervention for the treatment of hospitalized patients with COVID-19 infection. Trial registration: ClinicalTrials.govNCT04429854. Registered on 12 June 2020 - Retrospectively registered. [ABSTRACT FROM AUTHOR]

The research objective was to study the presence of DNA damages in haddock exposed to petrogenic or pyrogenic polyaromatic hydrocarbons (PAHs) from different sources: 1) extracts of oil produced water (PW), dominated by 2-ring PAHs; 2) distillation fractions of crude oil (representing oil-based drilling mud), dominated by 3-ring PAHs; 3) heavy pyrogenic PAHs, mixture of 4/5/6-ring PAHs. The biological effect of the different PAH sources was studied by feeding juvenile haddock with low doses of PAHs (0.3–0.7 mg PAH/kg fish/day) for two months, followed by a two-months recovery. In addition to the oral exposure, a group of fish was exposed to 12 single compounds of PAHs (4/5/6-ring) via intraperitoneal injection. The main endpoint was the analysis of hepatic and intestinal DNA adducts. In addition, PAH burden in liver, bile metabolites, gene and protein expression of CYP1A, GST activity, lipid peroxidation, skeletal deformities and histopathology of livers were evaluated. Juvenile haddock responded quickly to both intraperitoneal injection and oral exposure of 4/5/6-ring PAHs. High levels of DNA adducts were detected in livers three days after the dose of the single compound exposure. Fish had also high levels of DNA adducts in liver after being fed with extracts dominated by 2-ring PAHs (a PW exposure scenario) and 3-ring PAHs (simulating an oil exposure scenario). Elevated levels of DNA adducts were observed in the liver of all exposed groups after the 2 months of recovery. High levels of DNA adduct were found also in the intestines of individuals exposed to oil or heavy PAHs, but not in the PW or control groups. This suggests that the intestinal barrier is very important for detoxification of orally exposures of PAHs. [ABSTRACT FROM AUTHOR]

The purpose of this article is to present a new calibration procedure for spectroscopic measurements using an infrared focal plane array (IRFPA) spectrometer on highly opaque middle-wave infrared (MWIR) media. The procedure is based on the properties of the IRFPA camera and especially the integration time (IT), which is the main parameter that can be adjusted to control the sensitivity of the measurements. The goal of the paper is to experimentally validate this dependence with the direct reference intensity light coming out of the IR monochromator in order to predict the spectrum shape and intensity level in a range out of the camera saturation. This method allows determining spectrum used as background for transmittance calculation. It has been applied in the case of measurement of water transmittance, which is a highly opaque medium and whose measurement requires high ITs. The main result is the ability to take an IR spectroscopic imaging measurement through 300 µm of water and the determination of its transmittance with sufficient sensitivity due to the proposed calibration procedure. This procedure allows the possibility of transitory studies in heterogeneous aqueous media. [ABSTRACT FROM AUTHOR]

The synthesis of mycobacterial mycolic acid related wax esters and of their trehalose di- and mono-esters is described. The trehalose dimycolates (TDMs) synthesised activated bone marrow derived dendritic cells (BMDCs) in vitro more strongly than trehalose dibehenate or the trehalose monomycolates (TMMs). The inflammatory effects were similar to those of TDM from either a synthetic keto- or methoxy-mycolic acid, but somewhat stronger than those of a TDM from an α-mycolic acid. In vivo, the effects of one wax ester TDM were similar to those of the methoxy-MA and α-mycolic acid TDMs and trehalose dibehenate. [ABSTRACT FROM AUTHOR]

The rapid loss, fragmentation and degradation of tropical forests threaten the survival of many animal species. However, the way in which these phenomena affect animal health has been poorly explored, thus limiting the design of appropriate conservation strategies. To address this, here we identified using linear mixed models the effect of proximal (diet, activity pattern, hunting and logging) and distal (sum of the basal areas of fruiting-tree species [SBAFS], landscape forest cover and degree of forest fragmentation) variables over fecal glucocorticoid metabolite (fGCM) levels–hormones associated with animal health and fitness–of six groups of spider monkeys (Ateles geoffroyi) inhabiting six landscapes with different spatial structures in Mexico. Proximal variables showed a stronger predictive power over fGCMs than distal. In this sense, increases in travel time, the occurrence of hunting, and reductions in rest time and fruit consumption resulted in higher fGCM levels. Regarding distal variables, increases in SBAFS were negatively related to fGCM levels, thus suggesting that food scarcity increases stress hormone levels. Nevertheless, contrary to theoretical expectations, spider monkeys living in smaller tracts of forest spent less time travelling, but the same time feeding on fruit as those in more forested areas. The lower net energy return associated with this combination of factors would explain why, contrary to theoretical expectations, increased forest cover was associated with increased levels of fGCMs in these groups. Our study shows that, at least in the short term, spider monkeys in fragmented landscapes do not always present higher levels of stress hormones compared to those inhabiting continuous forest, and the importance of preserving fruit sources and controlling hunting for reducing the levels of stress hormones in free ranging spider monkeys. [ABSTRACT FROM AUTHOR]

The only tuberculosis vaccine currently available, bacille Calmette-Guérin (BCG) is a poor inducer of CD8+ T cells, which are particularly important for the control of latent tuberculosis and protection against reactivation. As the induction of strong CD8+ T cell responses is a hallmark of DNA vaccines, a combination of BCG with plasmid DNA encoding a prototype TB antigen (Ag85A) was tested. As an alternative animal model, pigs were primed with BCG mixed with empty vector or codon-optimized pAg85A by the intradermal route and boosted with plasmid delivered by intramuscular electroporation. Control pigs received unformulated BCG. The BCG-pAg85A combination stimulated robust and sustained Ag85A specific antibody, lymphoproliferative, IL-6, IL-10 and IFN-γ responses. IgG1/IgG2 antibody isotype ratio reflected the Th1 helper type biased response. T lymphocyte responses against purified protein derivative of tuberculin (PPD) were induced in all (BCG) vaccinated animals, but responses were much stronger in BCG-pAg85A vaccinated pigs. Finally, Ag85A-specific IFN-γ producing CD8+ T cells were detected by intracellular cytokine staining and a synthetic peptide, spanning Ag85A131-150 and encompassing two regions with strong predicted SLA-1*0401/SLA-1*0801 binding affinity, was promiscuously recognized by 6/6 animals vaccinated with the BCG-pAg85A combination. Our study provides a proof of concept in a large mammalian species, for a new Th1 and CD8+ targeting tuberculosis vaccine, based on BCG-plasmid DNA co-administration. [ABSTRACT FROM AUTHOR]

Fast-growing mycobacteria such as Mycobacterium sp. and Mycobacterium smegmatis degrade natural sterols. They are a model to study tuberculosis. Interestingly, M. smegmatis has been found in river effluents derived from paper production, and therefore, it would be important to gain further insight into its capacity to synthesize steroids that are potential endocrine disruptors affecting the development and reproduction of fishes. To our knowledge, the capacity of M. smegmatis to synthesize estrogens and even testosterone has not been previously reported. Therefore, the objective of this study was to investigate the capacity of M. smegmatis to synthesize in vitro testosterone and estrogens from tritiated precursors and to investigate the metabolic pathways involved. Results obtained by thin-layer chromatography showed that 3H-progesterone was transformed to 17OH-progesterone, androstenedione, testosterone, estrone, and estradiol after 6, 12, or 24 h of incubation. 3H-androstenedione was transformed into testosterone and estrogens, mainly estrone, and 3H-testosterone was transformed to estrone and androstenedione. Incubation with 3H-dehydroepiandrosterone rendered androstenediol, testosterone, and estrogens. This ability to transform less potent sex steroids like androstenedione and estrone into other more active steroids like testosterone and estradiol or vice versa suggests that M. smegmatis can influence the amount of self-synthesized strong androgens and estrogens and can transform those found in the environment. [ABSTRACT FROM AUTHOR]

Objective To determine the needle-to-nerve distances during electrical nerve location in dogs at different currents and pulse duration using a peripheral nerve stimulator ( PNS) under ultrasound control ( US), and the minimal electrical thresholds ( MET) necessary to obtain a motor response ( MR) after achieving needle-to-nerve contact. Study design Prospective in vivo experimental trial in a clinical setting Animals Thirty dogs, scheduled for locoregional anaesthesia of the sciatic nerve. Methods Needle-to-nerve distance was measured ultrasonographically after obtaining the MR of sciatic nerve with 2, 1 and 0.5 mA and pulse duration 0.1 ms (NS0.1). Thereafter the needle was placed in contact with the nerve and MET was determined. The procedure was repeated with 0.3 ms (NS0.3). Finally the needle was reintroduced to contact the sciatic nerve guided only by US, thus MET- US was determined. Data were analysed using Kruskal- Wallis or Mann- Whitney tests. Results Needle-to-nerve distances were greater when MR was obtained with 2 mA than with 1 and 0.5 mA at 0.1 and 0.3 ms. No significant differences were observed between the needle-to-nerve distances using 0.1 or 0.3 ms. The MET [median (range)] was 0.4 (0.18-1.3) mA in NS0.1, 0.32 (0.12-0.8) mA in NS0.3; while MET- US was 0.7 (0.32-1.5) mA. When the needle contacted the nerve, the MR achieved with currents below 0.3 mA was obtained in 17.2, 40 and 0% of cases using NS0.1, NS0.3 and US respectively. Conclusions and clinical relevance The electrical current necessary to obtain a MR decreased as the needle moved towards the nerve. However when the needle tip contacted the nerve, an MR with low current intensity could not be obtained. Thus the absence of motor response at currents below 0.3 mA cannot rule out needle-epineurium contact. When ultrasound is combined with PNS, it is more important to assess the correct needle position than searching for an MR at low currents. [ABSTRACT FROM AUTHOR]

Interleukin-17A ( IL-17A), a pro-inflammatory cytokine acting on neutrophil recruitment, is known to play an important role during Mycobacterium tuberculosis infection, but the role of IL-17 A receptor signalling in immune defence against this intracellular pathogen remains poorly documented. Here we have analysed this signalling using C57 BL/6 mice genetically inactivated in the IL-17 receptor A subunit ( IL-17 RA−/−). Although early after infection bacterial growth was controlled to the same extent as in wild-type mice, IL-17 RA−/− mice were defective in exerting long-term control of M. tuberculosis infection, as demonstrated by a progressively increasing pulmonary bacterial burden and shortened survival time. Compared with infected wild-type mice, IL-17 RA−/− mice showed impaired recruitment of neutrophils to the lungs at the early but not the late stage of infection. Pulmonary tumour necrosis factor-α, IL-6 and particularly IL-10 levels were decreased in the absence of IL-17 RA signalling, whereas IL-1β was increased. CD4+-mediated and γδ-mediated IL-17 A production was dramatically increased in IL-17 RA−/− mice (confirming part of their phenotype), whereas production of interferon-γ and expression of the bactericidal enzyme inducible nitric oxide synthase were not affected. Collectively, our data suggest that early but not late neutrophil recruitment is essential for IL-17 A-mediated long-term control of M. tuberculosis infection and that a functional interferon-γ response is not sufficient to control M. tuberculosis growth when the IL-17 RA pathway is deficient. As treatment of auto-immune diseases with anti- IL-17 A antibodies is actually being tested in clinical studies, our data suggest that caution should be taken with respect to possible reactivation of tuberculosis. [ABSTRACT FROM AUTHOR]

Objective To describe a new approach to block the femoral nerve and to evaluate the distribution of a dye injected into the psoas compartment using a new femoral nerve block approach; to assess its clinical application, when combined with a sciatic nerve block, for surgical anaesthesia/analgesia of the pelvic limb in dogs. Study design Prospective anatomical, research and clinical study. Animals Two dog cadavers; two dogs that had to be euthanized for reasons unrelated to this study, and 15 dogs undergoing pelvic limb orthopaedic surgery. Methods Phase 1: anatomical dissections were performed to determine a simple method to approach the femoral nerve within the psoas compartment. Phase 2: 0.1 mL kg−1 of a lidocaine-new methylene blue solution was injected bilaterally after successful electrolocation of the femoral nerve in two anaesthetized dogs. Colorant spread was evaluated through femoral nerve dissections after euthanasia. Phase 3: in 15 dogs undergoing pelvic limb orthopaedic surgery under light general anaesthesia with isoflurane, intra-operative analgesic effect (cardiovascular responses) and early post-operative pain score, of the novel femoral nerve block combined with a sciatic nerve block as the sole analgesic protocol, were evaluated. Results Phase 1: a needle inserted from the lateral aspect of the lumbar muscles, cranially to the iliac crest and with a 30-45° caudo-medial direction, reaches the femoral nerve in the caudal portion of the psoas compartment. Phase 2: Four femoral nerves were stained >2 cm. Phase 3: this novel lateral pre-iliac approach, combined with the sciatic nerve block, blunted the intra-operative cardiovascular response to surgical stimulation in 13 out of 15 anaesthetized dogs. In addition, rescue analgesia was not required in the early post-operative 2-hour period. Conclusion and clinical relevance The lateral pre-iliac femoral nerve block technique may provide adequate intra- and early post-operative pain relief in dogs undergoing pelvic limb surgery. [ABSTRACT FROM AUTHOR]