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Background: Leopardus geoffroyi is a Neotropical wild felid with wide distribution in the south of the South American continent. The objective was to investigate the skeletopy of the intumescentia lumbalis (IL) and conus medullaris (CM) from 11 specimens of L. geoffroyi collected dead on highways.Materials and Methods: The cadavers were fixed in formaldehyde solution and dissected to allow the dorsal exposure of IL and CM. The cranial and caudal limits were marked with radiopaque pins and radiographic projections were used to determine the skeletopy. The lengths of IL and CM were measured with a pachymeter.Results: In most specimens, the IL was located at the level of L4 and L5 vertebrae, although in 4 (1 male and 3 females) individuals its cranial limit was L3 and in 3 specimens (2 male and 1 female) the caudal limit was L6. The length of IL was 35.6 ± 6.7 mm. The CM had its base predominantly at the level of the L5 vertebra, although in some specimens the base was in L4 and in others in L6. The apex of the CM can be found since the lumbosacral junction until the level of the Cd2 vertebra. The CM measured 74.4 ± 14.3 mm.Conclusions: Based on the skeletopy, it can be suggested that epidural anaesthesia procedures in L. geoffroyi are safer with the introduction of the catheter through the sacrocaudal interarcual space, as recommended by some anaesthetists for the domestic cat. [ABSTRACT FROM AUTHOR]

Abstract Background Sevoflurane potently enhances glycine receptor currents and more modestly decreases NMDA receptor currents, each of which may contribute to immobility. This modest NMDA receptor antagonism by sevoflurane at a minimum alveolar concentration (MAC) could be reciprocally related to large potentiation of other inhibitory ion channels. If so, then reduced glycine receptor potency should increase NMDA receptor antagonism by sevoflurane at MAC. Methods Indwelling lumbar subarachnoid catheters were surgically placed in 14 anesthetized rats. Rats were anesthetized with sevoflurane the next day, and a pre-infusion sevoflurane MAC was measured in duplicate using a tail clamp method. Artificial CSF (aCSF) containing either 0 or 4 mg/mL strychnine was then infused intrathecally at 4 μL/min, and the post-infusion baseline sevoflurane MAC was measured. Finally, aCSF containing strychnine (either 0 or 4 mg/mL) plus 0.4 mg/mL dizocilpine (MK-801) was administered intrathecally at 4 μL/min, and the post-dizocilpine sevoflurane MAC was measured. Results Pre-infusion sevoflurane MAC was 2.26%. Intrathecal aCSF alone did not affect MAC, but intrathecal strychnine significantly increased sevoflurane requirement. Addition of dizocilpine significantly decreased MAC in all rats, but this decrease was two times larger in rats without intrathecal strychnine compared to rats with intrathecal strychnine, a statistically significant (P Conclusions Glycine receptor antagonism increases NMDA receptor antagonism by sevoflurane at MAC. The magnitude of anesthetic effects on a given ion channel may therefore depend on the magnitude of its effects on other receptors that modulate neuronal excitability.

Bradypus variegatus has unique anatomical characteristics, and many of its vascular and digestive tract aspects have yet to be clearly understood. This lack of information makes clinical diagnoses and surgical procedures difficult. The aim of this study was to evaluate the anatomical aspects of frozen and glycerinated corpses of B. variegatus using computed tomography (CT), emphasizing vascular and digestive contrast studies. Nine corpses that died during routine hospital were examined via CT in the supine position with scanning in the craniocaudal direction. In frozen cadavers, the contrast was injected into a cephalic vein after thawing and, subsequently, was administered orally. In addition to bone structures, CT allowed the identification of organs, soft tissues, and vascular structures in specimens. Visualization of soft tissues was better after contrast been administered intravenously and orally, even without active vascularization. Furthermore, the surfaces of the organs were highlighted by the glycerination method. With this technique, it was possible to describe part of the vascularization of the brachial, cervical, thoracic, and abdominal regions, in addition to highlighting the esophagus and part of the stomach. CT can be another tool for the evaluation of B. variegatus cadavers by anatomists or pathologists, contributing to the identification of anatomical structures.

The aim of this study was to evaluate the effects of pre-anesthetic use of clonidine on time-domain heart rate variability (HRV) and arterial blood pressure in healthy anesthetized dogs. Six healthy adult mixed-breed dogs were administered a clonidine (clonidine group, CLG) and 30 days later, a placebo (control group, CG) preanesthetic protocol, in addition to propofol, isoflurane, and an bolus of tramadol and the continuous infusion thereafter. The total time mean values of HRV meanNN, SDNN, SDANN, SDNNI, and rMSSD were higher in the CL group, as observed in some HRV variables on tramadol bolus time (T4), tramadol continuous infusion (T8), and tracheal extubation time (T10). No significant differences in arterial blood pressure were observed, however, two dogs had a second-degree atrioventricular block (Mobitz II) at the tramadol bolus time (T4). These results led us to conclude that the clonidine anesthetic protocol resulted in sympathetic outflow block and an increase in parasympathetic tone, without significant effects on blood pressure. Notably, cardiac electrical disturbance in two dogs in the CL group. Although the pre-anesthetic use of clonidine in dogs with fear-based behavioral problems should be considered, its association with tramadol should be avoided or carried out with caution owing to the existing cardiovascular risk., Competing Interests: Conflict of interests: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper., (Copyright Oliveira et al.)

Background: Symptomatic patients with severe brachymetatarsia are commonly treated with callus distraction using external mini-fixator (EF) or internal device (ID). This study points out advantages and limitations of both methods comparing clinical and radiographical parameters., Methods: Retrospective analysis of 21 metatarsal bones in twelve patients. Twelve metatarsals were treated with ID (Genos Mini), nine with EF (MiniRail)., Results: Mean lengthening distance was 17.3 mm using EF and 11.7 mm using ID (p = 0.016). Adverse results were observed in 89% of metatarsals treated with EF and in 33% treated with ID (p = 0.011). Postoperative surgical intervention was required in 33% using EF compared to 0% using ID (p = 0.031). Mean total German Foot Function Index (FFI-T) improved from 49 to 33 using EF and from 47 to 22 using ID (p < 0.001)., Conclusion: CD is a reliable surgical treatment for BMT. Surgeons should be aware of implant-related advantages and complications when counseling patients about treatment options., Competing Interests: Conflicts of interest and source of funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. The authors declare no conflicts of interest., (Copyright © 2022 European Foot and Ankle Society. Published by Elsevier Ltd. All rights reserved.)

Background: Different fertility control methods are investigated as a tool for population control of free-roaming animals. Chemical castration using calcium chloride has been widely studied over the years in males, but there are few studies related to its use in females. Therefore, we aimed to evaluate the local effects, as a potential chemosterilant, of two concentrations of calcium chloride diluted in 95% ethanol when administered by transcutaneous ultrasound-guided intraovarian injection in rats. In this study, 30 female Wistar rats were randomly divided into three treatment groups, which consisted of transcutaneous ultrasound-guided intraovarian injection of: 0.9% sodium chloride solution (GC); 10% calcium chloride diluted in 95% ethanol (G10); 20% calcium chloride diluted in 95% ethanol (G20). The animals were subdivided into two evaluation times, 15 days (n = 5 of each group) and 30 days (n = 5 of each group) after the intraovarian injection. The ovarian diameter was measured using ultrasound image prior and immediately after the injection and after the treatment period. Furthermore, animals' clinical evaluation, estrous cycles assessment, macroscopic examination of the abdominal cavity and histological evaluation of the ovaries were performed., Results: Ovarian ultrasound measurement revealed changes (p < 0.05) between ovarian diameters before and immediately after the injection in all treatments. Three animals in G20 had a small focal skin lesion at the injection site that evolved to total healing. Extended and abnormal estrous cycles were observed in G10 and G20. At gross examination, adhesions and ovarian cysts were noticed in both groups, G10 and G20. Also, the histopathology analysis revealed changes in ovarian architecture and vessel congestion in G10 and G20, but ovarian tissue damage was greater in the ovaries treated with the highest concentration (G20)., Conclusions: The results indicate that 20% calcium chloride diluted in 95% ethanol may be a potential agent for inducing sterilization in females and was possible to be minimally invasively delivered., (© 2022. The Author(s).)

The goal of this study was to develop a method for ultrasound-guided percutaneous intraovarian injection in Wistar rats. Intraovarian administration of chemicals or needle aspiration of the ovary has been undertaken in some species, including humans, equines, and bovines. In rodents, which are widely used in scientific research, a technique for intraovarian injection without surgical exposure of the organ has not been described. The current study standardized the procedure of ovarian percutaneous injection of 0.9% sodium chloride guided by ultrasound in rats. The ovaries were measured by ultrasound imaging before and immediately after injection and showed a significant increase in ovarian length but not width. No clinical abnormalities were detected within 15 d after injection. These findings indicate that the steps of ultrasound localization of the organ, digital restraint, and correct needle insertion achieved successful intraovarian administration of saline without invasive surgery. These results document the feasibility of ultrasound-guided intraovarian percutaneous injection in rats and may be useful for future research on female reproduction and chemical sterilization.

Background: Intraoperative fluids are still poorly studied in veterinary medicine. In humans the dosage is associated with significant differences in postoperative outcomes., Objectives: The aim of this study is to verify the influence of three different fluid therapy rates in dogs undergoing video-assisted ovariohysterectomy., Methods: Twenty-four female dogs were distributed into three groups: G5, G10, and G20. Each group was given 5, 10, and 20 mL·kg -1 ·h -1 of Lactate Ringer, respectively. This study evaluated the following parameters: central venous pressure, arterial blood pressure, heart rate, respiratory rate, temperature, acid-base balance, and serum lactate levels. Additionally, this study evaluated the following urinary variables: urea, creatinine, protein to creatinine ratio, urine output, and urine specific gravity. The dogs were evaluated up to 26 h after the procedure., Results: All animals presented respiratory acidosis during the intraoperative period. The G5 group evidenced intraoperative oliguria (0.80 ± 0.38 mL·kg -1 ·h -1 ), differing from the G20 group (2.17 ± 0.52 mL·kg -1 ·h -1 ) ( p = 0.001). Serum lactate was different between groups during extubation ( p = 0.036), with higher values being recorded in the G5 group (2.19 ± 1.65 mmol/L). Animals from the G20 group presented more severe hypothermia at the end of the procedure (35.93 ± 0.61°C) ( p = 0.032). Only the members of the G20 group presented mean potassium values below the reference for the species. Anion gap values were lower in the G20 group when compared to the G5 and G10 groups ( p = 0.017)., Conclusions: The use of lactated Ringer's solution at the rate of 10 mL·kg -1 ·h -1 seems to be beneficial in the elective laparoscopic procedures over the 5 or 20 mL·kg -1 ·h -1 rates of infusion., Competing Interests: The authors declare no conflicts of interest., (© 2021 The Korean Society of Veterinary Science.)

This study aimed to evaluate the ophthalmic parameters, morphometric features of corneal tissue, and arrangements of corneal stromal collagen fibers in crab-eating fox ( Cerdocyon thous ), a species of neotropical wild canid. We conducted the study on six juvenile crab-eating foxes (12 eyes), whilst 16 eyes were obtained post mortem from eight adult crab-eating foxes. The research was divided into two stages. In the first stage, eye anatomical characteristics, tear production (Schirmer 1 tear test, STT1), intraocular pressure (IOP), ocular echobiometry, and specular microscopy parameters related to morphology of corneal endothelium were studied in juvenile animals. In the second stage, morphometric features of corneal tissue (central corneal thickness [CCT] and corneal epithelium thickness) and arrangements of stromal collagen fibers were studied using eyes from adult animals. The main findings were that crab-eating fox eyes have vertical-slit pupils, holangiotic retina, and reference values (mean ± SD) of 13.37 ± 3.79 mm/min for STT1 and of 10.43 ± 3.84 mmHg for IOP. The ocular echobiometric features observed in crab-eating foxes are different from those reported for domestic dogs ( Canis familiaris ). Conversely, the corneal endothelial parameters are similar to those of domestic dogs. The CCT measured by tissue morphometry was 0.54 ± 0.06 mm, and the corneal epithelium thickness was 60.13 ± 8.71 µm. Mean coherency related to alignment of collagen fibers was 0.66 ± 0.12. The crab-eating fox cornea had predominantly thick collagen fibers. Crab-eating fox eyes have morphofunctional peculiarities. They resemble the eyes of domestic dogs in some aspects, but diverge in others.

Through the analysis of behavioural changes, this study demonstrates that methadone has behavioural, but not analgesic, effects on Oreochromis niloticus. It provides information that suggests the drug has sedative abilities, as the recovery time was shorter in the fish receiving methadone. Future research, with different doses and stimuli, is required to provide more information about analgesia., (© 2019 The Fisheries Society of the British Isles.)

Purpose: To evaluate acetylation of histone H3, chromatin remodeling, nuclear size and shape, DNA ploidy, and distribution of nucleolus organizing regions (NORs) in corneal epithelial and stromal cells of diabetic and nondiabetic rats., Methods: Diabetes was induced by a single intraperitoneal injection of alloxan. All diabetic rats (n = 20) included in the study had 4 weeks of moderate-to-severe hyperglycemia (plasma glucose levels >400 mg/dL). Acetylated histone H3 levels were quantified in corneal tissue using a colorimetric assay. Chromatin remodeling, nuclear sizes (area/perimeter) and shapes (circularity), and DNA ploidies were evaluated from Feulgen-stained tissue sections using video image analysis. Distributions of NORs were studied in tissue sections impregnated with silver ions. Ophthalmic clinical parameters, including corneal sensitivity, were investigated. Twenty nondiabetic rats were used as controls., Results: Acetylation of histone H3 was reduced in the corneas of the diabetic rats. Nuclei in corneal epithelial cells of diabetic rats compacted chromatin, increased in size, modified their shapes, and elevated DNA ploidy. The only nuclear change observed in the corneal stromal cells of diabetic rats was chromatin decompaction. The size of the silver-stained NOR did not differ between the study samples. The corneal sensitivity in diabetic rats was 51.8% lower than that in nondiabetic rats., Conclusions: The results of this study show that alloxan-induced diabetes altered the histone H3 acetylation pattern and compromised the chromatin supraorganization in corneal tissue/cells. Continued research is needed to understand the clinical and morphofunctional significance of changes in corneal cell nuclei of diabetic individuals.

Objective: To assess the temporal effects of a single fentanyl intravenous (IV) bolus on the minimum anesthetic concentration (MAC) of isoflurane in chickens and to evaluate the effects of this combination on heart rate (HR) and rhythm, systemic arterial pressures (sAP) and ventilation., Study Design: Prospective experimental trial., Animals: Seventeen adult chickens weighing 1.8±0.2 kg., Methods: Individual isoflurane MAC for 17 chickens was previously determined using the bracketing method. Chickens were anesthetized with isoflurane to evaluate the effects of a single IV fentanyl bolus (10 or 30 μg kg -1 ) on isoflurane MAC over time using the up-and-down method. Ventilation was controlled. The isoflurane MAC reduction was estimated by logistic regression at 5 and 15 minutes after fentanyl administration. In the second phase, seven chickens were anesthetized with isoflurane, and fentanyl was administered (30 μg kg -1 ) IV over 1 minute during spontaneous ventilation and HR and rhythm, sAP and ventilation variables were measured., Results: At 5 minutes after IV administration of fentanyl (10 or 30 μg kg -1 ), isoflurane MAC was significantly reduced by 17.6% (6.1-29.1%) [logistic regression estimate (95% Wald confidence interval)] and 42.6% (13.3-71.9%), respectively. Isoflurane MAC reduction at 15 minutes after IV administration of fentanyl (10 or 30 μg kg -1 ) was 6.2% (-0.6 to 12.9%) and 13.2% (-0.9 to 27.3%), respectively; however, this reduction was not significant. No clinically significant cardiopulmonary changes or arrhythmias were detected after the administration of fentanyl (30 μg kg -1 )., Conclusions and Clinical Relevance: Administration of a single fentanyl bolus induced a dose-dependent and short-lasting reduction in isoflurane MAC. The higher dose induced no significant cardiopulmonary depression in isoflurane-anesthetized chickens during spontaneous ventilation. In chickens anesthetized with isoflurane, the clinical usefulness of a single fentanyl bolus is limited by its short duration of effect., (Copyright © 2017 Association of Veterinary Anaesthetists and American College of Veterinary Anesthesia and Analgesia. Published by Elsevier Ltd. All rights reserved.)

Chrysocyon brachyurus (maned wolf) is the biggest South American canid and has a high frequency of dental injuries, both in the wild and in captivity. Thus, veterinary procedures are necessary to preserve the feeding capacity of hundreds of captive specimens worldwide. The aim of this study was to investigate the mandibular morphometry of the maned wolf with emphasis on the establishment of anatomic references for anesthetic block of the inferior alveolar and mental nerves. Therefore, 16 measurements in 22 mandibles of C. brachyurus adults were taken. For extraoral block of the inferior alveolar nerve at the level of the mandibular foramen, the needle should be advanced close to the medial face of the mandibular ramus for 11.4 mm perpendicular to the palpable concavity. In another extraoral approach, the needle may be introduced for 30.4 mm from the angular process at a 20-25° angle to the ventral margin. For blocking only the mental nerve, the needle should be inserted for 10 mm from ventral border, close to the labial surface of the mandibular body, at the level of the lower first premolar. The mandibular foramen showed similar position, size, and symmetry in the maned wolf specimens examined. Comparison of the data observed here with those available for other carnivores indicates the need to determine these anatomic references specifically for each species.

Purpose: To study the effects of topical administration of 1% morphine on corneal analgesia in rabbits submitted to lamellar keratectomy and to assess the expression of matrix metalloproteinase-1, metalloproteinase-2, metalloproteinase-9 (MMPs), type IV collagen, and interleukin-10 (IL-10) during the treatment., Methods: Morphine group (MG) received 50 μL of topical 1% morphine four times daily, while the control group received saline instead. Corneal touch threshold (CTT) and the wound area were assessed until corneal healing. Corneal samples were processed for routine histology, immunohistochemistry, zymography, and ELISA., Results: Following keratectomy, CTT increased significantly from 6 to 96 h time points. Mean corneal re-epithelization rate and scores of leukocyte infiltration did not differ significantly between treatment groups. Immunolabeling pattern for MMP-1, MMP-9, and type IV collagen was similar in both treatment groups. In the MG, zymography indicated significantly higher levels of active MMP-2 on days 6 and 12; and in the latent MMP-9, on days 3 and 6, and in the active MMP-9, on day 6. Latent MMP-2 and MMP-9, and active MMP-9 decreased to values close to those of healthy corneas on day 12, but levels of active MMP-2 remained significantly elevated in the MG. IL-10 levels measured on days 1-6 were reduced as compared to those of healthy corneal tissue and returned to levels close to those of healthy corneas on day 12., Conclusion: Topical morphine promoted corneal analgesia for up to 4 days and did not delay corneal re-epithelization. The re-establishment of MMPs and IL-10 to levels close to baseline values at the end of the study and the expression of type IV collagen in both groups reinforce that, with caution, 1% morphine can be used after lamellar keratectomy in rabbits., (© 2011 American College of Veterinary Ophthalmologists.)

Objective: To determine the minimum anesthetic concentration (MAC) for sevoflurane and measure the dose and temporal effects of butorphanol on the MAC for sevoflurane in guineafowl., Animals: 10 healthy adult guineafowl (Numida meleagris)., Procedures: Each bird was anesthetized with sevoflurane, and a standard bracketing method was used to measure the MAC in response to a noxious electrical stimulus. Subsequently, conditions were adjusted so that each bird was anesthetized with sevoflurane at a fraction of its respective MAC (eg, 0.7 times the MAC for that bird). Butorphanol tartrate (2 mg/kg, IV) was administered, and a noxious stimulus was applied every 15 minutes until the bird moved in response. The reduction in MAC was estimated with logistic regression by use of a standard quantal method. After an interval of ≥ 1 week, the MAC reduction experiment was repeated with an increased butorphanol dosage (4 mg/kg)., Results: Individual mean ± SE MAC for sevoflurane was 2.9 ± 0.1%. At 15 minutes after administration of 2 mg of butorphanol/kg, estimated reduction in the MAC for sevoflurane was 9 ± 3%. At 15 and 30 minutes after administration of 4 mg of butorphanol/kg, estimated reduction in the MAC for sevoflurane was 21 ± 4% and 11 ± 8%, respectively., Conclusions and Clinical Relevance: In guineafowl, the MAC for sevoflurane was similar to values reported for other species. Increasing the butorphanol dosage decreased the MAC for sevoflurane, but the effect was small and of short duration for dosages up to 4 mg/kg.

To evaluate the cardiorespiratory changes induced by isoflurane (ISO) anesthesia in the crested caracara (Caracara plancus), eight crested caracaras that weighed 1.0 kg (range 0.9-1.1 kg) were the subjects for the study. The birds were anesthetized by face mask with ISO for brachial artery catheterization. After recovery, anesthesia was re-induced and maintained with ISO with spontaneous ventilation. Electrocardiography, direct systolic arterial blood pressure (SAP), diastolic arterial blood pressure (DAP), mean arterial blood pressure (MAP), respiratory rate (RR), end-tidal carbon dioxide (P(ET)CO2), and cloacal temperature (T degrees C) were measured before induction (baseline, under physical restraint) and after 5, 10, 15, 20, 25, 30, 35, and 40 min of ISO anesthesia. Arterial blood samples were collected for blood gas analysis at baseline, 10, 25, and 40 min. No cardiac arrhythmias were observed in the present study. RR, SAP, DAP, MAP, T degrees C and pH decreased from baseline values, whereas arterial partial pressures of oxygen and carbon dioxide, bicarbonate concentration, and P(ET)CO2 were significantly higher than baseline. Apnea was not observed in any bird. ISO anesthesia is suitable for use in healthy members of this species despite the moderate cardiovascular and respiratory depression produced.

Objective: To evaluate the cardiorespiratory changes induced by sevoflurane (SEV) anesthesia in the crested caracara (Caracara plancus)., Study Design: Prospective experimental trial., Animals: Eight crested caracaras (Caracara plancus) weighing 1.0 (0.9-1.1) kg were used for the study., Methods: The birds were anesthetized by face mask with isoflurane for brachial artery catheterization. After recovery, anesthesia was re-induced with 6% SEV via face mask. After induction, a noncuffed endotracheal tube was placed and anesthesia was maintained with SEV (3.5% end-tidal) in oxygen (1 L minute(-1)) using an Ayre's T-piece nonrebreathing circuit, with spontaneous ventilation. Electrocardiography (ECG), direct systolic, diastolic and mean arterial blood pressure (SAP, DAP, and MAP), respiratory rate (f(R)), end-tidal carbon dioxide (Pe'CO(2)), and cloacal temperature (T degrees C) were measured before induction (baseline - under physical restraint) and after 5, 10, 15, 20, 25, 30, 35 and 40 minutes of SEV anesthesia. Arterial blood samples were collected for gas analysis at baseline and then at 10, 25 and 40 minutes., Results: No ventricular arrhythmias were observed in the present study. Respiratory rate, SAP, DAP, MAP, T degrees C and pH decreased from pre-induction values, while arterial partial pressures of oxygen and carbon dioxide, bicarbonate concentration, and Pe'CO(2) were significantly higher than baseline. None of the birds were apneic., Conclusion and Clinical Relevance: Sevoflurane anesthesia is suitable for use in healthy members of this species, despite the moderate cardiovascular and respiratory depression produced.

Objective: to evaluate the interrelationships between cerebral and systemic hemometabolic alterations in patients with severe traumatic brain injury managed according to a standardized therapeutic protocol., Design: prospective, interventional study in patients with traumatic coma., Setting: a general Intensive Care Unit in a teaching hospital., Patients and Methods: twenty-seven patients (21M e 6F), aging 14 - 58 years, with severe acute brain trauma, presenting with three to eight points on the Glasgow Coma Scale, were prospectively evaluated according to a cumulative protocol for the management of acute intracranial hypertension, where intracranial pressure (ICP) and cerebral extraction of oxygen (CEO2) were routinely measured. Hemometabolic interrelationships involving mean arterial pressure (MAP), ICP, arterial carbon dioxide tension (PaCO2), CEO2, cerebral perfusion pressure (CPP) and systemic extraction of oxygen (SEO2) were analyzed., Interventions: routine therapeutic procedures., Results: no correlation was found between CEO2 and CPP (r = -0.07; p = 0.41). There was a significant negative correlation between PaCO2 and CEO2 (r = -0.24; p = 0.005) and a positive correlation between SEO2 and CEO2 (r = 0.24; p = 0.01). The mortality rate in this group of patients was 25.9% (7/27)., Conclusion: 1) CPP and CEO2 are unrelated; 2) CEO2 and PaCO2 are closely related; 3) during optimized hyperventilation, CEO2 and SEO2 are coupled.