Beneficial effects of recombinant CER-001 high-density lipoprotein infusion in sepsis: results from a bench to bedside translational research project.

BACKGROUND: Sepsis is characterized by a dysregulated immune response and metabolic alterations, including decreased high-density lipoprotein cholesterol (HDL-C) levels. HDL exhibits beneficial properties, such as lipopolysaccharides (LPS) scavenging, exerting anti-inflammatory effects and providing endothelial protection. We investigated the effects of CER-001, an engineered HDL-mimetic, in a swine model of LPS-induced acute kidney injury (AKI) and a Phase 2a clinical trial, aiming to better understand its molecular basis in systemic inflammation and renal function. METHODS: We carried out a translational approach to study the effects of HDL administration on sepsis. Sterile systemic inflammation was induced in pigs by LPS infusion. Animals were randomized into LPS (n = 6), CER20 (single dose of CER-001 20 mg/kg; n = 6), and CER20 × 2 (two doses of CER-001 20 mg/kg; n = 6) groups. Survival rate, endothelial dysfunction biomarkers, pro-inflammatory mediators, LPS, and apolipoprotein A-I (ApoA-I) levels were assessed. Renal and liver histology and biochemistry were analyzed. Subsequently, we performed an open-label, randomized, dose-ranging (Phase 2a) study included 20 patients with sepsis due to intra-abdominal infection or urosepsis, randomized into Group A (conventional treatment, n = 5), Group B (CER-001 5 mg/kg BID, n = 5), Group C (CER-001 10 mg/kg BID, n = 5), and Group D (CER-001 20 mg/kg BID, n = 5). Primary outcomes were safety and efficacy in preventing AKI onset and severity; secondary outcomes include changes in inflammatory and endothelial dysfunction markers. RESULTS: CER-001 increased median survival, reduced inflammatory mediators, complement activation, and endothelial dysfunction in endotoxemic pigs. It enhanced LPS elimination through the bile and preserved liver and renal parenchyma. In the clinical study, CER-001 was well-tolerated with no serious adverse events related to study treatment. Rapid ApoA-I normalization was associated with enhanced LPS removal and immunomodulation with improvement of clinical outcomes, independently of the type and gravity of the sepsis. CER-001-treated patients had reduced risk for the onset and progression to severe AKI (stage 2 or 3) and, in a subset of critically ill patients, a reduced need for organ support and shorter ICU length of stay. CONCLUSIONS: CER-001 shows promise as a therapeutic strategy for sepsis management, improving outcomes and mitigating inflammation and organ damage. TRIAL REGISTRATION: The study was approved by the Agenzia Italiana del Farmaco (AIFA) and by the Local Ethic Committee (N° EUDRACT 2020-004202-60, Protocol CER-001- SEP_AKI_01) and was added to the EU Clinical Trials Register on January 13, 2021.

Effects of GLUBRAN-2 on the Burst Pressure of Jejunal Loops Thermofused With Vessel Sealing Devices.

BACKGROUND: Off-label use of radiofrequency vessel sealing devices for intestinal thermofusion has been reported as an alternate approach for closing the small and large intestines. The study aimed to evaluate if reinforcing the thermofusion line with a modified N-butyl-2-CyanoAcrylate and methacryloxysulpholane produced improved burst pressure values in ex vivo swine jejunal loops. MATERIALS AND METHOD: A suture-less full-thickness jejunal biopsy was performed with different radiofrequency vessel-sealing devices (Ligasure 5 mm: RFVS-1; Atlast 10 mm: RFVS-2; Cayman Maryland: RFVS-3), and reinforcement with modified cyanoacrylate Glubran-2 (G2) at the thermofusion defect was applied. Burst pressure(BP) values were compared with a control group, wherein a cold blade was utilized to obtain the biopsy, followed by the closing of the jejunum with seven Gambee sutures. RESULTS: Seventy (n = 70) jejunal loop samples were distributed into the experimental groups.The RFVS-1 and -2 groups exhibited BP values similar to those of the suture group. The RFVS-3 group showed significantly lower BP values (P < 0.05) than the suture group. Conversely, in the groups wherein G2 was applied, all BP values were comparable to those of the suture group. BP test in the RFVS-3G2 group showed significantly (P < 0.05) higher values in the group using the same instrument without the glue (RFVS-3). CONCLUSIONS: G2 has been shown to improve the BP on the defects created by instruments that are not completely efficient in intestinal thermofusion and sealing. This experimental model showed that the performance of full-thickness biopsies with RFVS devices and reinforcement with G2 provide feasible and promising results.

Laparoscopic salpingectomy in Papio hamadryas for birth control in captivity.

OBJECTIVE: To evaluate the feasibility of laparoscopic salpingectomy in baboons (Papio hamadryas). We hypothesized that laparoscopic salpingectomy could be performed in baboon species within a reasonable amount of surgical time, with minor complications occurring at low rates. STUDY DESIGN: Case series and technique description. ANIMALS: Sixteen baboons (n = 16). METHODS: The surgical procedures were performed using the 3-port technique, with 5 mm instruments and a telescope placed at the umbilical and hypogastric regions. A salpinx dissection was performed, using a radiofrequency bipolar vessel sealing device, from the fimbriae to the uterine attachments. We evaluated the surgical duration, learning curve, and intraoperative and early postoperative complications. RESULTS: Ten adult and 6 subadult baboons with a mean weight of 9.32 kg, a range of 4-14.2 kg, and a standard deviation (SD) of 3.09 kg were included in the study. The total duration of surgery was 28.75 min (range, 16-50 min; SD, 9.60 min). The installation phase was completed in a mean time of 7.68 min (range, 3-15 min; SD, 3.43 min), and the time to complete the salpingectomy of both salpinges was 9.68 min (range, 4-20 min; SD, 3.97 min). No complications were observed in the postoperative period. CONCLUSION: Laparoscopic salpingectomy in Papio hamadryas was feasible, with an acceptable surgical time, low invasiveness, and only minor technical perioperative complications. CLINICAL SIGNIFICANCE: Laparoscopic salpingectomy could be a viable and safe therapeutic option in nonhuman primate birth-control programs.

Absorbable fixation straps for laparoscopic gastropexy in dogs.

OBJECTIVE: To evaluate the feasibility and efficacy of laparoscopic absorbable fixation straps (AFS) for laparoscopic gastropexy in dogs. STUDY DESIGN: Cadaveric and prospective clinical study. ANIMALS: Five dog cadavers for the cadaveric study; 12 dogs for the clinical study. METHODS: The pyloric antrum was affixed to the abdominal wall laparoscopically by applying a series of straps. The cadaveric study assessed potential challenges during the procedure and stomach mucosal penetration. For the clinical study, the total duration of surgery, time to complete the gastropexy, and the number of straps used were recorded. Ultrasound evidence of adhesion, complications, and weight were monitored at 7, 30, and 90 days after surgery. Owner satisfaction was evaluated at the 6-month follow-up. RESULTS: The total duration of surgery was 25.8 minutes (range, 19-39; SD, 6.7), and the time to complete the gastropexy was 10.1 minutes (range, 7-19; SD, 3.9). The linear regression analysis revealed an inverse correlation between the time to complete the gastropexy and the order of the surgeries (r2  = 0.75, P < .05). No complications were recorded. Ultrasound examination was used to confirm gastropexy at all follow-ups. CONCLUSION: Laparoscopic gastropexy with AFS was performed in both cadavers and clinical animals with minimal complications. Persistent adhesion was demonstrated during ultrasound evaluations and in one postmortem evaluation. CLINICAL SIGNIFICANCE: This novel laparoscopic technique can be employed safely, effectively, and reasonably quickly, and the learning curve is expected to be relatively short.

Clinical efficacy of dexmedetomidine combined with lidocaine for femoral and sciatic nerve blocks in dogs undergoing stifle surgery.

OBJECTIVE: To evaluate the effects of dexmedetomidine administered perineurally or intramuscularly (IM) on sensory, motor function and postoperative analgesia produced by lidocaine for sciatic and femoral nerve blocks in dogs undergoing unilateral tibial tuberosity advancement surgery. STUDY DESIGN: Prospective, blinded, clinical study. ANIMALS: A group of 30 dogs. METHODS: Dogs were anaesthetized with acepromazine, propofol and isoflurane in oxygen/air. Electrolocation-guided femoral and sciatic nerve blocks were performed: group L, 0.15 mL kg-1 2% lidocaine (n = 10); group LDloc, lidocaine and 0.15 µg kg-1 dexmedetomidine perineurally (n = 10); group LDsys, lidocaine and 0.3 µg kg-1 dexmedetomidine IM (n = 10). After anaesthesia, sensory blockade was evaluated by response to forceps pinch on skin innervated by the saphenous/femoral, common fibular and tibial nerves. Motor blockade was evaluated by observing the ability to walk and proprioception. Analgesia was monitored with Short Form of Glasgow Composite Pain Scale for up to 4 hours after extubation. Methadone IM was administered as rescue analgesia. Data were analysed by linear mixed effect models and Kaplan-Meier test (p < 0.05). RESULTS: Median duration of the sensory blockade for all nerves was longer (p < 0.001) for group LDloc than for groups L and LDsys and was longer (p = 0.0011) for group LDsys than for group L. Proprioception returned later (p < 0.001) for group LDloc [285 (221-328) minutes] compared with group L [160 (134-179) minutes] or LDsys [195 (162-257) minutes]. Return of the ability to walk was similar among all groups. Dogs in group LDloc required postoperative rescue analgesia later (p = 0.001) than dogs in groups LDsys and L. CONCLUSIONS AND CLINICAL RELEVANCE: Dexmedetomidine administered perineurally with lidocaine prolonged sensory blockade and analgesia during the immediate postoperative period. Systemic dexmedetomidine also prolonged the sensory blockade of perineural lidocaine.

Definition and clinical evaluation of a recruiting airway pressure based on the specific lung elastance in anesthetized dogs.

OBJECTIVE: To determine the specific lung elastance (SEL) in anesthetized dogs and to evaluate the efficacy of a SEL-based recruiting airway pressure (RPaw) at improving global and regional lung aeration. STUDY DESIGN: Retrospective and prospective clinical study. ANIMALS: A total of 28 adult dogs were included in the retrospective study and six adult dogs in the prospective study. METHODS: Retrospective study: SEL and SEL-based RPaw were determined using previously published data. In mechanically ventilated dogs undergoing thoracic computed tomography (CT), SEL was calculated as ΔPL/(VT/EELV), where ΔPL is the driving transpulmonary pressure, VT is the tidal volume and EELV is the end-expiratory lung volume. The ratio of lung to respiratory system elastance (EL/Ers) was determined. SEL and EL/Ers were used to calculate the SEL-based RPaw. Prospective study: dogs underwent thoracic CT at end-expiration and at end-inspiration using the SEL-based RPaw, and global and regional aeration was determined. For analysis of regional aeration, lungs were divided into cranial, intermediate and caudal regions. Regional compliance was also calculated. A p value <0.05 was considered significant. RESULTS: The SEL and EL/Ers were 12.7 ± 3.1 cmH2O and 0.54 ± 0.07, respectively. The SEL-based RPaw was 29.1 ± 7.6 cmH2O. In the prospective study, the RPaw was 28.2 ± 1.3 cmH2O. During RPaw, hyperinflation increased (p = 0.0003) whereas poorly aerated (p < 0.0001) and nonaerated (p = 0.01) tissue decreased. Normally aerated tissue did not change (p = 0.265). Regional compliance was higher in the intermediate (p = 0.0003) and caudal (p = 0.034) regions compared with the cranial region. Aeration did not differ between regions (p > 0.05). CONCLUSIONS AND CLINICAL RELEVANCE: An SEL-based RPaw reduces poorly and nonaerated lung tissue in anesthetized dogs. In nonsurgical anesthetized dogs, an RPaw near 30 cmH2O is effective at improving lung aeration.

LPS-Binding Protein Modulates Acute Renal Fibrosis by Inducing Pericyte-to-Myofibroblast Trans-Differentiation through TLR-4 Signaling.

During sepsis, the increased synthesis of circulating lipopolysaccharide (LPS)-binding protein (LBP) activates LPS/TLR4 signaling in renal resident cells, leading to acute kidney injury (AKI). Pericytes are the major source of myofibroblasts during chronic kidney disease (CKD), but their involvement in AKI is poorly understood. Here, we investigate the occurrence of pericyte-to-myofibroblast trans-differentiation (PMT) in sepsis-induced AKI. In a swine model of sepsis-induced AKI, PMT was detected within 9 h from LPS injection, as evaluated by the reduction of physiologic PDGFRß expression and the dysfunctional α-SMA increase in peritubular pericytes. The therapeutic intervention by citrate-based coupled plasma filtration adsorption (CPFA) significantly reduced LBP, TGF-ß, and endothelin-1 (ET-1) serum levels, and furthermore preserved PDGFRß and decreased α-SMA expression in renal biopsies. In vitro, both LPS and septic sera led to PMT with a significant increase in Collagen I synthesis and α-SMA reorganization in contractile fibers by both SMAD2/3-dependent and -independent TGF-ß signaling. Interestingly, the removal of LBP from septic plasma inhibited PMT. Finally, LPS-stimulated pericytes secreted LBP and TGF-ß and underwent PMT also upon TGF-ß receptor-blocking, indicating the crucial pro-fibrotic role of TLR4 signaling. Our data demonstrate that the selective removal of LBP may represent a therapeutic option to prevent PMT and the development of acute renal fibrosis in sepsis-induced AKI.

Effects of positive end-expiratory pressure alone or an open-lung approach on recruited lung volumes and respiratory mechanics of mechanically ventilated horses.

OBJECTIVE: To evaluate the effects of positive end-expiratory pressure (PEEP) alone and PEEP preceded by lung recruitment manoeuvre (LRM) on lung volumes and respiratory system mechanics in healthy horses undergoing general anaesthesia. STUDY DESIGN: Controlled, prospective clinical study. ANIMALS: A group of 15 horses undergoing arthroscopy. METHODS: Following anaesthetic induction, initial ventilatory settings were: tidal volume 15 mL kg-1, inspiratory:expiratory ratio 1:2, respiratory rate to maintain end-tidal CO2 between 5.3-6.6 kPa (40-50 mmHg). The following settings were implemented sequentially: zero PEEP (ZEEP); PEEP 10 cmH2O (PEEP); LRM (50 cmH2O for 20 seconds) followed by 10 cmH2O of PEEP (LRM + PEEP). Static compliance (Cst), driving pressure, delta end-expiratory (ΔEELV) and recruited lung volumes (RLV) were obtained 30 minutes after initiating each ventilatory strategy. Data were analyzed with paired t test or analysis of variance followed by Tukey's post hoc test. Data are shown as mean ± standard deviation; p < 0.05 was considered significant. RESULTS: PEEP induced ΔEELV of 6.68 ± 3.36 mL kg-1; ΔEELV during LRM + PEEP was 14.28 ± 5.59 mL kg-1 (p < 0.0001). The RLV was greater during the LRM + PEEP phase (12.30 ± 5.85 mL kg-1) than during PEEP (4.47 ± 3.97 mL kg-1; p < 0.0001). The Cst was unchanged from ZEEP to PEEP (0.75 ± 0.21 and 0.85 ± 0.22 mL cmH2O-1 kg-1, respectively, p = 0.36) but increased using LRM + PEEP (1.11 ± 0.25 mL cmH2O-1 kg-1, p = 0.0004). Driving pressure was lower during LRM + PEEP than during PEEP and ZEEP (16 ± 2, 19 ± 2 and 21 ± 4 cmH2O, respectively, p < 0.0001). CONCLUSIONS AND CLINICAL RELEVANCE: Unlike PEEP alone, PEEP preceded by LRM increased RLV and Cst and reduced driving pressure in horses under anaesthesia.

Perioperative analgesic effects of an ultrasound-guided transversus abdominis plane block with a mixture of bupivacaine and lidocaine in cats undergoing ovariectomy.

OBJECTIVE: To evaluate the perioperative analgesic effects of a transversus abdominis plane (TAP) block with a mixture of lidocaine and bupivacaine administered to cats undergoing ovariectomy. STUDY DESIGN: Controlled, randomized, prospective, blinded clinical study. ANIMALS: A group of 20 healthy cats. METHODS: Robenacoxib (2 mg kg-1) was administered subcutaneously 0.5 hour before intramuscular (IM) administration of ketamine (5 mg kg-1), methadone (0.1 mg kg-1) and dexmedetomidine (0.01 mg kg-1). General anesthesia was induced with intravenous (IV) propofol and maintained with isoflurane. An ultrasound-guided TAP block was performed by injecting 0.5% bupivacaine (0.2 mL kg-1) diluted in a total volume of 1.5 mL 2% lidocaine bilaterally (TAP group, n = 10) or the same volume of saline solution bilaterally in controls (CTR group, n = 10). During surgery, a 20% increase in heart rate and respiratory frequency was treated with IV fentanyl (0.001 mg kg-1). Before premedication and at 1, 2, 3, 4, 6, 8, 12, 16, 20 and 24 hours after extubation, pain was assessed with a simple descriptive pain scale, that ranged from 0 (no pain) to 4 (intense pain). For pain scores ≥3, IM methadone (0.1 mg kg-1) was administered. Data were analyzed with the Friedman or the analysis of variance (anova) test, and p < 0.05 was considered statistically significant. RESULTS: Only two cats in the CTR group were administered one dose of fentanyl during surgery. At 2, 6, 8, 12, 16, 20 and 24 hours after surgery, the pain score was higher in the CTR group. A mean dose of 0.5 ± 0.2 mg kg-1 methadone was administered to all cats in the CTR groups within 24 hours. Methadone was not administered to the TAP group (pain score < 3). CONCLUSIONS AND CLINICAL RELEVANCE: Ultrasound-guided TAP block can be a reliable adjunctive technique, providing analgesia for up to 24 hours in cats undergoing ovariectomy.

An alveolar recruitment maneuver followed by positive end-expiratory pressure improves lung function in healthy dogs undergoing laparoscopy.

OBJECTIVE: To evaluate the effects of an alveolar recruitment maneuver (ARM) followed by 5 cmH2O positive end-expiratory pressure (PEEP) in dogs undergoing laparoscopy. STUDY DESIGN: Prospective, randomized clinical study. ANIMALS: A group of 20 dogs undergoing laparoscopic ovariectomy. METHODS: Dogs were sedated with acepromazine and methadone intramuscularly; anesthesia was induced with propofol intravenously and maintained with inhaled isoflurane. The following baseline ventilatory setting (BVS) was administered: tidal volume of 12 mL kg-1, inspiratory to expiratory ratio of 1:2, inspiratory pause 25% of inspiratory time, no PEEP and a respiratory rate to maintain end-tidal carbon dioxide tension between 5.3 and 7.3 kPa. Then, 10 minutes after the pneumoperitoneum, 10 dogs (RM) underwent a sustained inflation ARM followed by BVS plus 5 cmH2O PEEP, while 10 dogs (NO-RM) were left with BVS throughout the procedure. Gas exchange and respiratory system mechanics were evaluated before the pneumoperitoneum (PPpre), before ARM (PP10), 30 minutes later (PP30) and 20 minutes after pneumoperitoneum discontinuation (PPpost20). Data were analyzed using anova (p < 0.05). RESULTS: The Fshunt at PP30 and PPpost20 was lower (p < 0.001) in the RM (2.3 ± 2.2 and 4.7 ± 3.7%) than in the NO-RM (5.2 ± 2.1 and 11.1 ± 5.2%), and PaO2 at PP30 and PPpost20 was higher (p < 0.001) in the RM (67.3 ± 4.2 and 60.1 ± 9.4 kPa) than in the NO-RM (50.2 ± 7.4 and 45.5 ± 11.1 kPa). Static compliance of the respiratory system at PP30 and PPpost20 was greater (p < 0.001) in the RM (2.4 ± 0.2 and 2.1 ± 0.4 mL cmH2O-1 kg-1) than in the NO-RM (0.9 ± 0.4 and 1.2 ± 0.2 mL cmH2O-1 kg-1). CONCLUSIONS AND CLINICAL RELEVANCE: In dogs undergoing laparoscopy, ARM followed by 5 cmH2O PEEP improves gas exchange and respiratory system mechanics.

WHICH AIRWAY PRESSURE SHOULD BE APPLIED DURING BREATH-HOLD IN DOGS UNDERGOING THORACIC COMPUTED TOMOGRAPHY?

This randomized controlled trial study aimed to identify the optimal positive pressure (PP) level that can clear atelectasis while avoiding pulmonary hyperinflation during the breath-hold technique in dogs undergoing thoracic computed tomography (CT). Sixty dogs affected by mammary tumors undergoing thoracic CT for the screening of pulmonary metastases were randomly assigned to six groups with different levels of PP during the breath-hold technique: 0 (control), 5 (PP5), 8 (PP8), 10 (PP10), 12 (PP12), and 15 (PP15) cmH2 O. The percentage of atelectatic lung region was lower in the PP10 (3.7 ± 1.1%; P = 0.002), PP12 (3.4 ± 1.3%; P = 0.0001), and PP15 (2.8 ± 0.9%; P = 0.006) groups than in the control group (5.0 ± 2.3%), and the percentage of poorly aerated lung region was lower in the PP8 (15.1 ± 2.6%; P = 0.0009), PP10 (13.0 ± 2.0 %; P = 0.002), PP12 (13.0 ± 2.2 %; P = 0.0002), and PP15 (11.1 ± 1.9%; P = 0.0002) groups than in the control group (19.8 ± 5.0). The percentage of normally aerated lung region, however, was higher in the PP10 (79.7 ± 4.1%; P = 0.005), PP12 (79.8 ± 5.1%; P = 0.0002), and PP15 (80.2 ± 4.9%; P = 0.002) groups than in the control group (73.4 ± 6.6%). A PP of 10-12 cmH2 O during the breath-hold technique should be considered to improve lung aeration during a breath-hold technique in dogs undergoing thoracic CT.

Low respiratory rate plus minimally invasive extracorporeal Co2 removal decreases systemic and pulmonary inflammatory mediators in experimental Acute Respiratory Distress Syndrome.

OBJECTIVE: The Acute Respiratory Distress Syndrome Network protocol recommends limiting tidal volume and plateau pressure; it also recommends increasing respiratory rate to prevent hypercapnia. We tested a strategy that combines the low tidal volume with lower respiratory rates and minimally invasive CO2 removal. SUBJECTS: Ten lung-damaged pigs (instilled hydrochloride). INTERVENTIONS: Two conditions randomly applied in a crossover fashion: the Acute Respiratory Distress Syndrome Network protocol and the Acute Respiratory Distress Syndrome Network protocol plus lower respiratory rate plus minimally invasive Co2 removal. A similar arterial Co2 partial pressure was targeted in the two conditions. MEASUREMENTS AND MAIN RESULTS: Physiological parameters, computed tomography scans, plasma and bronchoalveolar lavage concentrations of interleukin-1ß, interleukin-6, interleukin-8, interleukin-10, interleukin-18, and tumor necrosis factor-α. During the lower respiratory rate condition, respiratory rate was reduced from 30.5 ± 3.8 to 14.2 ± 3.5 (p < 0.01) breaths/min and minute ventilation from 10.4 ± 1.6 to 4.9 ± 1.7 L/min (p < 0.01). The extracorporeal device removed 38.9% ± 6.1% (79.9 ± 18.4 mL/min) of CO2 production. During the lower respiratory rate condition, interleukin-6, interleukin-8, and tumor necrosis factor-α concentrations were significantly lower in plasma; interleukin-6 and tumor necrosis factor-α concentrations were lower in bronchoalveolar lavage, whereas the concentrations of the other cytokines remained unchanged. CONCLUSION: The strategy of lower respiratory rate plus minimally invasive extracorporeal CO2 removal was feasible and safe and, as compared with the Acute Respiratory Distress Syndrome Network protocol, reduced the concentrations of some, but not all, of the tested cytokines without affecting respiratory mechanics, gas exchange, and hemodynamics.

Testicular expression of heat SHOCK proteins 60, 70, and 90 in cryptorchid horses.

Heat shock proteins are the most evolutionarily conserved protein families induced by stressors including hyperthermia. In the context of pathologies of the male reproductive tract, cryptorchidism is the most common genital defect that compromises the reproductive potential of the male because it induces an increase in intratesticular temperature. In equine species, cryptorchidism affects almost 9 % of newborns and few studies have been carried out on the molecular aspects of the retained testis. In this study, the expression pattern of HSP60, 70, and 90 in abdominal and inguinal testes, in their contralateral descended normally testes, and in testes of normal horses were investigated by Western blot and immunohistochemistry. The histomorphological investigation of retained and scrotal testes was also investigated. The seminiferous epithelium of the retained testes showed a vacuolized appearance and displayed a completely blocked spermatogenesis for lacking meiotic and spermiogenetic cells. On the contrary, the contralateral scrotal testes did not show morphological damage and the seminiferous epithelium displayed all phases of the spermatogenetic cycle as in the normal testes. The morphology of Leydig cells was not affected by the cryptorchid state. Western blot and immunohistochemistry evidenced that equine testis (both scrotal and retained) expresses the three investigated HSPs. More in detail, the Western blot evidenced that HSP70 is the more expressed chaperone and that together with HSP90 it is highly expressed in the retained gonad (P < 0.05). The immunohistochemistry revealed the presence of the three HSPs in the spermatogonia of normal and cryptorchid testes. Spermatogonia of retained testes showed the lowest expression of HSP60 and the highest expression of HSP90. Spermatocytes, spermatids of scrotal testes, and the Sertoli cells of retained and scrotal testes did not display HSP60 whereas expressed HSP70 and HSP90. These two proteins were also localized in the nucleus of the premeiotic cells. The Leydig cells displayed the three HSPs with the higher immunostaining of HSP70 and 90 in the cryptorchid testes. The results indicate that the heat stress condition occurring in the cryptorchid testis influences the expression of HSPs.

The abundance and localization of heat shock proteins (HSP)-60, -70, and -90 in the oviductal ampulla of hamadryas baboon (Papio hamadryas) during the menstrual cycle.

The presence of HSPs in female reproductive and their relationship with the steroid hormone fluctuation have been reported in several mammals but not in non-human primates. The present research dealt with the oviductal expression and localization of the more studied HSPs (60, 70, and 90) as well as the morphological changes in the Hamadryas baboon (Papio hamadryas) during the follicular, preovulatory, and luteal phases of the menstrual cycle. Therefore, western blots, histomorphological, and immunohistochemical analyses were carried out. The results of western blot analysis displayed the lowest HSP expression in the luteal phase. The histomorphology showed that the mucosal epithelium consisted of undifferentiated cuboidal cells in follicular and luteal phases and well-distinguishable columnar ciliated and non-ciliated cells during the preovulatory phase. Immunohistochemistry evidenced that the mucosal epithelium contained cytoplasmic and nuclear HSP60, 70, and 90 immunostaining in the follicular and luteal phases. During the preovulatory phase, the non-ciliated cells showed: (i) cytoplasmic HSP60; (ii) nuclear and cytoplasmic HSP90. Ciliated cells showed cytoplasmic and ciliary HSP70 and ciliary HSP90. The stromal cells and myocytes of muscular layer displayed a decreased cytoplasmic HSP60 in the preovulatory phase and nuclear and low cytoplasmic HSP70 throughout the menstrual cycle. Nuclear HSP90 decreased in ampulla stromal cells and the follicular phase myocytes. These findings indicate that the expression pattern of HSP60,70, and 90 is related to the morphofunctional features of the baboon oviductal ampulla during the menstrual cycle and could represent a referent point for further studies in the oviduct of Primates.

Respiratory Effects of Continuous Positive Airway Pressure Administered during Recovery from General Anesthesia in Brachycephalic Dogs.

This study aimed to evaluate the benefits of applying 5 cmH2O of CPAP using a pediatric helmet during the recovery phase from general anesthesia in brachycephalic dogs. Brachycephalic dogs undergoing various surgical procedures were included in this study, and a total of 64 subjects were randomly assigned to receive either standard oxygen supplementation (NO-CPAP group) or oxygen supplementation combined with CPAP (CPAP group). This study evaluated arterial blood pH, blood gas partial pressures of O2 and CO2, arterial blood O2 saturation, and related parameters during recovery. The dogs were monitored, and helmet tolerance was assessed using predefined criteria. Of the initially assessed 69 dogs, 64 were enrolled: 32 in the CPAP group and 32 in the NO-CPAP group. Fifteen dogs in the NO-CPAP group were excluded based on predetermined criteria. The CPAP group showed significant improvements in PaO2, PaO2/FiO2, P(A-a)O2, F-Shunt, and respiratory rate compared with the NO-CPAP group (p < 0.001). The incidence of reintubation and helmet intolerance was higher in the NO-CPAP group (18% and 15.6%, respectively) than in the CPAP group (0%). This study highlights the potential benefits of incorporating CPAP, delivered through a pediatric helmet, in the perioperative management of brachycephalic dogs.

Recruitment-to-inflation ratio reflects the impact of peep on dynamic lung strain in a highly recruitable model of ARDS.

BACKGROUND: The recruitment-to-inflation ratio (R/I) has been recently proposed to bedside assess response to PEEP. The impact of PEEP on ventilator-induced lung injury depends on the extent of dynamic strain reduction. We hypothesized that R/I may reflect the potential for lung recruitment (i.e. recruitability) and, consequently, estimate the impact of PEEP on dynamic lung strain, both assessed through computed tomography scan. METHODS: Fourteen lung-damaged pigs (lipopolysaccharide infusion) underwent ventilation at low (5 cmH2O) and high PEEP (i.e., PEEP generating a plateau pressure of 28-30 cmH2O). R/I was measured through a one-breath derecruitment maneuver from high to low PEEP. PEEP-induced changes in dynamic lung strain, difference in nonaerated lung tissue weight (tissue recruitment) and amount of gas entering previously nonaerated lung units (gas recruitment) were assessed through computed tomography scan. Tissue and gas recruitment were normalized to the weight and gas volume of previously ventilated lung areas at low PEEP (normalized-tissue recruitment and normalized-gas recruitment, respectively). RESULTS: Between high (median [interquartile range] 20 cmH2O [18-21]) and low PEEP, median R/I was 1.08 [0.88-1.82], indicating high lung recruitability. Compared to low PEEP, tissue and gas recruitment at high PEEP were 246 g [182-288] and 385 ml [318-668], respectively. R/I was linearly related to normalized-gas recruitment (r = 0.90; [95% CI 0.71 to 0.97) and normalized-tissue recruitment (r = 0.69; [95% CI 0.25 to 0.89]). Dynamic lung strain was 0.37 [0.29-0.44] at high PEEP and 0.59 [0.46-0.80] at low PEEP (p < 0.001). R/I was significantly related to PEEP-induced reduction in dynamic (r = - 0.93; [95% CI - 0.78 to - 0.98]) and global lung strain (r = - 0.57; [95% CI - 0.05 to - 0.84]). No correlation was found between R/I and and PEEP-induced changes in static lung strain (r = 0.34; [95% CI - 0.23 to 0.74]). CONCLUSIONS: In a highly recruitable ARDS model, R/I reflects the potential for lung recruitment and well estimates the extent of PEEP-induced reduction in dynamic lung strain.

Pressure-Sensitive Walkway System for Evaluation of Lameness in Dogs Affected by Unilateral Cranial Cruciate Ligament Rupture Treated with Porous Tibial Tuberosity Advancement.

The aim of this study was to objectively evaluate lameness in dogs affected by a unilateral cranial cruciate ligament rupture (CrCLR) treated with porous tibial tuberosity advancement before surgery and at three different timepoints after surgery, using the GAITRite® system (version 4.9Wr), a pressure-sensitive walkway system that is able to calculate several spatiotemporal gait parameters simultaneously for each limb. The dogs walked on the pressure-sensitive walkway before (T0) and 30 (T1), 90 (T2), and 120 (T3) days after surgery. Pressure measurements (gait lameness score and total pressure index %) were collected for S (treated with porous TTA) and C (healthy contralateral limb) at T0, T1, T2, and T3 and statistically evaluated. An ANOVA test was performed to compare the data, and a value of p < 0.05 was considered significant. Twenty dogs (n = 20) of various common breeds and ages with CrCLR were enrolled in the study. The results showed that there was a statistically significant difference in the GAIT4Dog® lameness score (GLS) and TPI% between S and C for each timepoint. Statistically significant differences in the GLS and TPI% between S at T0 and S at T2 and between S at T0 and S at T3 (p < 0.001) were found. The results showed that there was a statistically significant difference in the GAIT4Dog® lameness score (GLS) and TPI% between S and C for each timepoint. Statistically significant differences in the GLS and TPI% between S at T0 and S at T2 and between S at T0 and S at T3 were found. The GLS and TPI% increased statistically significantly from 90 days after surgery compared to the preoperative measurements. Moreover, comparing the GLS and TPI% between the treated limb and the control limb showed that a statistically significant difference remained at each timepoint.

Use of Laryngeal Mask and Anesthetic Management in Hamadryas Baboons (Papio hamadryas) Undergoing Laparoscopic Salpingectomy-A Case Series.

The study aims to describe the anesthetic and airway management of baboons (Papio hamadryas) undergoing laparoscopic salpingectomy with a laryngeal mask airway (LMA) device. Eleven baboons received tiletamine-zolazepam and medetomidine; anesthesia was induced with propofol. An LMA was positioned for oxygen and isoflurane administration in spontaneous respiration. Heart rate (HR), mean arterial pressure (MAP), respiratory rate (RR), end tidal carbon dioxide (EtCO2), minute volume (MV), and peripheral hemoglobin oxygen saturation (SpO2) were recorded before (PREPP) and immediately after abdomen insufflation (PP1), at 10 (PP2), 20 (PP3), and 30 (PP4) minutes during pneumoperitoneum, and after (POSTPP) pneumoperitoneum. The respiratory rate was significantly higher at all times compared to PREPP. The end tidal carbon dioxide concentration was significantly higher at PP2, PP3, PP4, and POSTPP, compared to the previous times. The higher values for RR and EtCO2 were registered at PP4: 22.7 (95% CI 17.6-27.8) breaths/min and 57.9 (95% CI 51.9-63.8) mmHg, respectively. The minute volume was significantly higher at PP4 and POSTPP compared to the other times. The higher value for MV was registered at POSTPP (269.1 (95% CI 206.1-331.8) mL/kg/min). This protocol is suitable for baboons undergoing laparoscopic salpingectomy. The LMA was easy to insert and allowed for good ventilation, gas exchange, and delivery of the anesthetic in spontaneous breathing baboons.

Ex-Vivo Evaluation of "First Tip Closing" Radiofrequency Vessel Sealing Devices for Swine Small Intestinal Transection.

This study compared burst pressure (BP), number of activations, and histological assessment of ex vivo swine small intestine loops transected by stapler, a single fulcrum radiofrequency vessel sealing (RFVS) device, and the newly-developed jaws RFVS. Fifty (n = 50) 20 cm long jejunal loops were randomly assigned to be transected with RFVS devices and linear stapler (Caiman5, Caiman Maryland, Caiman12, Ligasure Atlas, and Stapler group as control respectively). Caiman5, Caiman12 and stapler required only one activation to complete the sealing. The mean BP in Caiman5 and Caiman Maryland groups were significantly lower (p < 0.05) than the S group as control and the other RFVS devices studied. RFVS Caiman12 and Ligasure Atlas produced mean BP values that were close to the Control and did not differ between them. The lumen was totally closed in the Caiman12 and Ligasure Atlas groups. The findings of this investigation were promising; we discovered that Caiman12 and Ligasure Atlas produce comparable mechanical capabilities as well as stapled intestinal closure, however Caiman12 need a single activation to complete the transection.

Modification of Morphology and Glycan Pattern of the Oviductal Epithelium of Baboon Papio hamadryas during the Menstrual Cycle.

The mammalian oviduct is a highly specialized structure where fertilization and early embryonic development occur. Its mucosal epithelium is involved in maintaining and modulating a dynamic intraluminal fluid. The oviductal epithelium consists of ciliated and non-ciliated (secretory) cells whose differentiation and activity are sex hormone-dependent. In this study, we investigated for the first time both the morphology and the glycan composition of baboon oviductal epithelium during the menstrual cycle. Oviducts were laparoscopically removed from 14 healthy adult female Papio hamadryas whose menstrual cycle phase was assessed based on the sex hormone levels and the vaginal cytology features. Histological investigations were carried out on fimbriae, infundibulum, ampulla, and isthmus separately fixed in 4% (v/v) paraformaldehyde, embedded in paraffin wax, and stained with hematoxylin-eosin for morphological analyses and using a panel of nine fluorescent lectins for glycoconjugate characterization. The histomorphological analysis revealed that in the entire oviduct (i) the ciliated and non-ciliated cells were indistinguishable during the follicular and luteal phases, whereas they were highly differentiated during the preovulatory phase when the non-ciliated cells exhibited apical protrusions, (ii) the epithelium height was significantly higher in the preovulatory phase compared to other menstrual phases, and (iii) the number of ciliated cells significantly (p ≤ 0.05) increased from the fimbriae to the infundibulum and progressively reduced in the other oviductal segments with the lower presence of ciliated cells in the isthmus. The glycan characterization revealed a complex and region-specific composition during the different phases of the menstrual cycle. It can be summarized as follows: (i) high-mannosylated N-linked glycans (Con A reactivity) were present throughout the oviductal epithelium during the entire menstrual cycle and characteristically in the apical protrusions of non-ciliated cells of the ampulla during the preovulatory phase; (ii) sialoglycans with α2,3-linked sialic acids (MAL II binding) were expressed along the entire oviductal surface only during the preovulatory phase, whereas α2,6-linked ones (SNA affinity) were also detected in the surface of the luteal phase, although during the preovulatory phase they were characteristically found in the glycocalyx of the isthmus cilia, and O-linked sialoglycans with sialic acids linked to Galßl,3GalNAc (T antigen) (KsPNA) and terminal N-acetylgalactosamine (Tn antigen) (KsSBA) were found in the entire oviductal surface during all phases of the menstrual cycle; (iii) GalNAc terminating O-linked glycans (HPA staining) were mainly expressed in the entire oviducts of the luteal and preovulatory phases, and characteristically in the apical protrusions of the isthmus non-ciliated cells of the preovulatory phase; and (iv) fucosylated glycans with α1,2-linked fucose (LTA reactivity) occurred in the apical surface of fimbriae during the luteal phase, whereas α1,3/4-linked fucose (UEA I binders) were present in the apical protrusions of the ampulla non-ciliated cells and in the apical surface of isthmus during the preovulatory phase as well as in the isthmus apical surface of follicular-phase oviducts. These results demonstrate for the first time that morphological and glycan changes occur in the baboon oviductal epithelium during the menstrual cycle. Particularly, the sex hormone fluctuation affects the glycan pattern in a region-specific manner, probably related to the function of the oviductal segments. The findings add new data concerning baboons which, due to their anatomical similarity to humans, make an excellent model for female reproduction studies.