• Title/Summary/Keyword: 동물 모델

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The Whole Extract of Enterococcus faecalis Has Suppressive Effect on the Allergic Responses in Asthmatic Mouse Model (천식 마우스 모델의 알러지 반응에서 Enterococcus faecalis 전체 추출물의 억제 효과)

  • Chang, Jeong Hyun;Yang, EunJu;Yu, Sun Nyoung;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1168-1175
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    • 2017
  • Probiotics are usually defined as intestinal bacteria that provide healthy benefit to the host and may offer new therapeutic materials for the treatment of inflammatory diseases. Lactobacillus, Bifidobacterium and Enterococcus are known as typical probiotics. But, these bacteria have mostly a weak viability and thus decreased probiotics-mediated effects in the intestinal tract. Asthma is an inflammatory airway disease, which is characterized by the releases of inflammatory mediators including cytokine and IgE. They are mainly associated with the recruitment, activation and disregulation of specific inflammatory cells, especially mast cells, monocytes, T cells, eosinophils and neutrophils in asthma. We performed these studies as in vitro and in vivo test the human inflammatory cell lines and ovalbumin (OVA)-induced asthma mouse model. And then the inhibitory effects of Enterococcus faecalis whole extract on inflammatory responses were examined. For our examinations, the E. faecalis whole extract (Ef extract) was acquired from whole bacteria of E. faecalis using freeze/thawing after ultrasonication method. As results, OVA-mediated THP-1 cell viability was decreased by the treatment of Ef extract. In the asthmatic mouse model, Ef extract inhibited the infiltration of inflammatory cells into the inflammatory sites and blood. This whole extract may have anti-asthmatic effects associated with the regulation of IL-5 and IgE expression. It may also be a promising candidate in anti-allergic medicine for the treatment of asthma.

Inhibitory Effects of Schisandrae Fructus Ethanol Extract on the Production of Matrix Metalloproteinases in in vitro and in vivo Osteoarthritis Models (In vitro 및 in vivo 퇴행성관절염 모델에서 오미자 에탄올 추출물에 의한 matrix metalloproteinases의 생성 억제)

  • Jeong, Jin-Woo;Lee, Hye Hyeon;Kim, Hong Jae;Lee, Ki Won;Kim, Ki Young;Kim, Sung Goo;Hong, Su-Hyun;Kim, Bum Hoi;Park, Cheol;Choi, Yung Hyun
    • Journal of Life Science
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    • v.27 no.10
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    • pp.1207-1214
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    • 2017
  • Schisandrae Fructus (SF), the fruit of Schisandra chinensis (Turcz.) Baill., is widely used in traditional medicine for the treatment of a number of chronic diseases. SF extracts have been recently reported to attenuate the inflammatory responses in SW1353 human chondrocyte cells in in vitro and monosodium iodoacetate (MIA)-induced cartilage degradation in in vivo osteoarthritis (OA) models. However, their protective and therapeutic potentials against OA in primary culture chondrocytes and animal models remain unclear. Therefore, we investigated the effects of the ethanol extract of SF on the activity of matrix metalloproteinases (MMPs), biomarkers for diagnosis of OA, on interleukin $(IL)-1{\beta}-induced$ primary cultured rat cartilage chondrocytes and MIA-induced osteoarthritis in a rat model. Our data indicated that SF treatment significantly reduced the mRNA expression and enzyme activity of MMP-1, -3 and -13 in $IL-1{\beta}-induced$ primary cultured rat cartilage chondrocytes. The chondro-protective effects of SF were then analyzed in a rat OA model using a single intra-articular injection of MIA in the right knee joint. According to our results, the elevated levels of MMP-1 and -3 were markedly ameliorated by SF administration. Collectively, these findings indicate that SF could be a candidate for the treatment of OA.

Effect of Agaricus blazei β-Glucan and Egg Shell Calcium Complex on Bone Metabolism in Ovariectomized Rats (난소절제 흰쥐에서 신령버섯의 β-Glucan과 난각 Ca 복합체가 골 대사에 미치는 효과)

  • Noh, Kyung-Hee;Jang, Ji-Hyun;Kim, Jin-Ju;Park, Cherl-Woo;Kim, Jeong-Ok;Kim, Jae-Cherl;Song, Young-Sun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.35 no.10
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    • pp.1363-1370
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    • 2006
  • This study was designed to evaluate the effect of Agaricus blazei $\beta-glucan$ and egg shell calcium complex on bone metabolism in ovariectomized (OVX) rats. Forty Sprague-Dewley female rats, 10 weeks of age $(248{\pm}1.7g)$, were divided into 4 groups and fed on the experimental diets for 6 weeks: sham operated control treated with normal diet containing 0.5% calcium (Sham-C), OVX-control treated with normal diet containing 0.5% calcium (OVX-C), $OVX-\beta-glucan$ group treated with $\beta-glucan$ diet containing 0.5% calcium (OVX-G), and $OVX-\beta-glucan$ egg shell calcium complex treated with $OVX-\beta-glucan$ egg shell calcium complex containing 0.5% calcium (OVX-GE). Bone weight of femur was higher in the OVX-GE group than in the other OVX groups. Bone mineral density of femur was significantly different (p<0.05) among the experimental groups and showed the highest level in the OVX-GE group. Calcium absorption rate and retention were higher in the $\beta-glucan$ supplement groups than in the other groups (p<0.05). Alkaline phosphatase activities and osteocalcin levels of serum showed lower in the $\beta-glucan$ supplement groups than in the OVX-C group. Deoxypyridinoline crosslink values of urine, indicator of bone absorption, showed the lowest in the OVX-GE group. The $\beta-glucan$ supplemented groups had a lower bone resorption ratio than in the OVX-C group. We concluded that bioavailability of calcium is higher in $\beta-glucan$ supplement groups compared to those in OVX rats. From the above results, these findings suggest the possibility of using $\beta-glucan$ egg shell calcium complex as a functional food material related to bone metabolism, even though there is no significant difference between the groups of $\beta-glucan$ and $\beta-glucan-egg$ shell calcium complex supplementation.

Effects of Coculture on Development of Biopsied Mouse Embryos as a Preclinical Model for Preimplantation Genetic Diagnosis of Human Embryos (생쥐 모델을 이용한 배아의 할구 생검법과 할구가 생검된 배아의 배양시 공배양 효과에 관한 연구: 인간에서의 착상 전 유전진단 기술 개발을 위한 동물실험 모델의 개발)

  • Kim, S.H.;Ryu, B.Y.;Jee, B.C.;Choi, S.M.;Kim, H.S.;Pang, M.G.;Oh, S.K.;Suh, C.S.;Choi, Y.M.;Kim, J.G.;Moon, S.Y.;Lee, J.Y.;Chae, H.D.;Kim, C.H.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.1
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    • pp.9-20
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    • 1999
  • The genetic defects in human gametes and embryos can cause adverse effects on overall reproductive events. Biopsy of embryos for preimplantation genetic diagnosis (PGD) offers a new possibility of having children free of the genetic disease. In addition, advanced embryo culture method may enhance the effectiveness of embryo biopsy for the practical application of PGD. This experimental study was undertaken to evaluate the effects of coculture on the development in vitro of biopsied mouse embryos as a preclinical model for PGD of human embryos. Embryos were obtained after in vitro fertilization (IVF) from F1 hybrid mice (C57BLfemale/CBAmale). Using micromanipulation, 1, 2, 3 or 4 blastomeres of 8-cell stage embryos were aspirated through a hole made in the zona pellucida by zona drilling (ZD) with acidic Tyrode's solution (ATS). After biopsy of blastomeres, embryos were cultured in vitro for 110 hours in Ham's F-10 supplemented with 0.4% BSA or cocultured on the monolayer of Vero cells in the same medium. The frequence of blastocyst formation were recorded, and the embryos beyond blastocyst stage were stained with 10% Giemsa to count the total number of nuclei in each embryo. There was no significant difference in the blastocyst formation between the zona intact control group and the zona drilling (ZD) only, or biopsied groups. The hatching rate of all the treatment groups except 4/8 group was significantly higher than that of control group. In all the treatment groups, there was a significant reduction in the mean cell number of embryos beyond blastocyst stage ($50.2{\pm}14.0$ in control group vs. $41.2{\pm}7.9$ in ZD, $39.3{\pm}8.8$ in 7/8, $29.7{\pm}6.4$ in 6/8, $25.1{\pm}5.7$ in 5/8, and $22.1{\pm}4.3$ in 4/8 groups, p<0.05). When the same treatments were followed by coculture with Vero cells, a similar pattern was seen in the blastocyst formation and the hatching rate. However, in all the treatment groups, there was a significant increase in the mean cell number of embryos beyond blastocyst stage with coculture, compared with the parallel groups without coculture. In the cleavage rate of biopsied blastomeres cultured for 110 hours after IVF, there was no significant difference between coculture and non-coculture groups (87.2% vs. 78.7%). However, the mean cell number of embryos developed from the biopsied blastomeres was significantly higher in coculture group ($11.5{\pm}4.7\;vs.\;5.9{\pm}1.9$, p<0.05). In conclusion, biopsy of mouse embryos after ZD with ATS is a safe and highly efficient method for PGD, and coculture with Vero cells showed a positive effect on the development in vitro of biopsied mouse embryos and blastomeres as a preclinical model for PGD of human embryos.

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The Effect of Cultured Perichondrial Cell Sheet Covered Highly Active Engineered Cartilage: in vivo Comparative Assessment (배양연골막이 피복된 고효능 인공연골의 생체내 효과)

  • Park, Se-Il;Moon, Young-Mi;Jeong, Jae-Ho;Jang, Kwang-Ho;Ahn, Myun-Hwan
    • Journal of Veterinary Clinics
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    • v.28 no.5
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    • pp.486-496
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    • 2011
  • A special mesenchymal tissue layer called perichondrium has a chondrogenic capacity and is a candidate tissue for engineering of cartilage. To overcome limited potential for chondrocyte proliferation and re-absorption, we studied a method of cartilage tissue engineering comprising chondrocyte-hydrogel pluronic complex (CPC) and cultured perichondrial cell sheet (cPCs) which entirely cover CPC. For effective cartilage regeneration, cell-sheet engineering technique of high-density culture was used for fabrication of cPCs. Hydrogel pluronic as a biomimetic cell carrier used for stable and maintains the chondrocytes. The human cPCs was cultured as a single layer and entirely covered CPC. The tissue engineered constructs were implanted into the dorsal subcutaneous tissue pocket on nude mice (n = 6). CPC without cPCs were used as a controls (N = 6). Engineered cartilage specimens were harvested at 12 weeks after implantation and evaluated with gross morphology and histological examination. Biological analysis was also performed for glycosaminoglycan (GAG) and type II collagen. Indeed, we performed additional in vivo studies of cartilage regeneration using canine large fullthickness chondrial defect model. The dogs were allocated to the experimental groups as treated chondrocyte sheets with perichondrial cell sheet group (n = 4), and chondrocyte sheets only group (n = 4). The histological and biochemical studies performed 12 weeks later as same manners as nude mouse but additional immunofluorescence study. Grossly, the size of cartilage specimen of cPCs covered group was larger than that of the control. On histological examination, the specimen of cPCs covered group showed typical characteristics of cartilage tissue. The contents of GAG and type II collagen were higher in cPCs covered group than that of the control. These studies demonstrated the potential of such CPC/cPCs constructs to support chondrogenesis in vivo. In conclusion, the method of cartilage tissue engineering using cPCs supposed to be an effective method with higher cartilage tissue gain. We suggest a new method of cartilage tissue engineering using cultured perichondrial cell sheet as a promising strategy for cartilage tissue reconstruction.

Ameliorative Effects of Soybean Leaf Extract on Dexamethasone-Induced Muscle Atrophy in C2C12 Myotubes and a C57BL/6 Mouse Model (콩잎 추출물의 근위축 개선 효과)

  • Hye Young Choi;Young-Sool Hah;Yeong Ho Ji;Jun Young Ha;Hwan Hee Bae;Dong Yeol Lee;Won Min Jeong;Dong Kyu Jeong;Jun-Il Yoo;Sang Gon Kim
    • Journal of Life Science
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    • v.33 no.12
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    • pp.1036-1045
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    • 2023
  • Sarcopenia, a condition characterized by the insidious loss of skeletal muscle mass and strength, represents a significant and growing healthcare challenge, impacting the mobility and quality of life of aging populations worldwide. This study investigated the therapeutic potential of soybean leaf extract (SL) for dexamethasone (Dexa)-induced muscle atrophy in vitro and in an in vivo model. In vitro experiments showed that SL significantly alleviated Dexa-induced atrophy in C2C12 myotube cells, as evidenced by preserved myotube morphology, density, and size. Moreover, SL treatment significantly reduced the mRNA and protein levels of muscle RING-finger protein-1 (MuRF1) and muscle atrophy F-box (MAFbx), key factors regulating muscle atrophy. In a Dexa-induced atrophy mouse model, SL administration significantly inhibited Dexa-induced weight loss and muscle wasting, preserving the mass of the gastrocnemius and tibialis anterior muscles. Furthermore, mice treated with SL exhibited significant improvements in muscle function compared to their counterparts suffering from Dexa-induced muscle atrophy, as evidenced by a notable increase in grip strength and extended endurance on treadmill tests. Moreover, SL suppressed the expression of muscle atrophy-related proteins in skeletal muscle, highlighting its protective role against Dexa-induced muscle atrophy. These results suggest that SL has potential as a natural treatment for muscle-wasting conditions, such as sarcopenia.

Habitat characteristics and prediction of potential distribution according to climate change for Macromia daimoji Okumura, 1949 (Odonata: Macromiidae) (노란잔산잠자리(Macromia daimojiOkumura, 1949)의 서식지 특성 및 기후변화에 따른 잠재적 분포 예측)

  • Soon Jik Kwon;Hyeok Yeong Kwon;In Chul Hwang;Chang Su Lee;Tae Geun Kim;Jae Heung Park;Yung Chul Jun
    • Journal of Wetlands Research
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    • v.26 no.1
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    • pp.21-31
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    • 2024
  • Macromia daimoji Okumura, 1949 was designated as an endangered species and also categorized as Class II Endangered wildlife on the International Union for Conservation of Nature (IUCN) Red List in Korea. The spatial distribution of this species ranged within a region delimited by northern latitude from Sacheon-si(35.1°) to Yeoncheon-gun(38.0°) and eastern longitude from Yeoncheon-gun(126.8°) to Yangsan-si(128.9°). They generally prefer microhabitats such as slowly flowing littoral zones of streams, alluvial stream islands and temporarily formed puddles in the sand-based lowland streams. The objectives of this study were to analyze the similarity of benthic macroinvertebrate communities in M. daimoji habitats, to predict the current potential distribution patterns as well as the changes of distribution ranges under global climate change circumstances. Data was collected both from the Global Biodiversity Information Facility (GBIF) and by field surveys from April 2009 to September 2022. We adopted MaxEnt model to predict the current and future potential distribution for M. daimoji using downloaded 19 variables from the WorldClim database. The differences of benthic macroinvertebrate assemblages in the mainstream of Nakdonggang were smaller than those in its tributaries and the other streams, based on the surrounding environments and stream sizes. MaxEnt model presented that potential distribution displayed high inhabiting probability in Nakdonggang and its tributaries. Applying to the future scenarios by Intergovernmental Panel on Climate Change (IPCC), SSP1 scenario was predicted to expand in a wide area and SSP5 scenario in a narrow area, comparing with current potential distribution. M. daimoji is not only directly threatened by physical disturbances (e.g. river development activities) but also vulnerable to rapidly changing climate circumstances. Therefore, it is necessary to monitor the habitat environments and establish conservation strategies for preserving population of M. daimoji.

The Relationship Between the NF-${\kappa}B$ Activity and Anti-inflammatory Action of Surfactant in the Acute Lung Injury of Rats (백서의 급성폐손상에서 surfactant의 항염증작용과 호중구의 NK-${\kappa}B$ 활성과의 관계)

  • An, Chang-Hyeok;Cha, Young-Joo;Lee, Kyoung-Hee;Yoo, Chul-Gyu;Lee, Byoung-Jun;Jeong, Do-Young;Lee, Sang-Hoon;Shin, Jong-Wook;Kim, Jae-Yeol;Park, In-Won;Choi, Byoung-Whui
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.5
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    • pp.519-529
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    • 2002
  • Background : The therapeutic effects of surfactants on acute lung injury derive not only from their recruiting action on collapsed alveoli but also from their anti-inflammatory action in the alveolar sapce. This study evaluated the anti-inflammatory action of a surfactant in an acute lung injury model of rats by neutrophils were recollected from the BAL fluid and the NF-${\kappa}B$ activity of the neutrophilic nuclear protein was evaluated. Methods : Male Sprague-Dawley rats weighing approximately 300 gram were divided into 3 groups, which consisted of 6 rats respectively. In the control group, normal saline(3ml/kg) was instilled into the trachea twice with 30 minute interval. In two other groups, acute lung injury was induced by the intra-tracheal instillation of LPS(5mg/kg). Thirty minutes later, either a surfactant(ST group; 30mg/kg) or normal saline(NT group: 3ml/kg) was instilled via the trachea. Twenty-four hours after the LPS instillation, the BAL fluid was retrieved to measure the WBC count and cytokine(IL-$1{\beta}$ and IL-6) levels. The neutrophils were isolated from the BAL fluid and the nuclear protein was extracted to evaluate the NF-${\kappa}B$ activity using a eletrophoretic mobility shift assay(EMSA). Results : The WBC count of the BAL fluid of the ST group($3,221{\pm}1,914{\times}10^3/{\mu}l$) was higher than that of the control group($356{\pm}275{\times}10^3/{\mu}l$)(p<0.05) and lower than that of the NT group($5,561{\pm}1,757{\times}10^3/{\mu}l$)(p<0.05)). The BAL fluid level of IL-$1{\beta}$ from the NT group($2,064{\pm}1,082pg/ml$) was higher than those of the ST group($360{\pm}234pg/ml$)(p<0.05) and the control group(0pg/ml)p<0.05) and control group($49{\pm}62pg/ml$)(p<0.05). The NF-${\kappa}B$ activity of the neutrophilic nuclear protein in the ST group and NT group was similar. Conclusion : The surfactant, attenuates the alveolar inflammation in the acute lung injury of rats model. However, its anti-inflammatory action does no't appear to be mediated by the inhibition of NF-${\kappa}B$ activity.

Immunohistochemical Localization of NMDA Receptor in the Auditory Brain Stem of Postnatal 7, 16 Circling Mouse (생후 7일, 16일된 circling mouse 청각 뇌줄기에서 N-메틸-D 아스파르트산염 수용체(NMDA receptor)에 대한 면역염색학적 분포)

  • Choi, In-Young;Park, Ki-Sup;Kim, Hye-Jin;Maskey, Dhiraj;Kim, Myeung-Ju
    • Applied Microscopy
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    • v.40 no.2
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    • pp.53-64
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    • 2010
  • Glutamate receptors may play a critical role in the refinement of developing synapses. The lateral superior olivary nucleus (LSO)-medial nucleus of trapezoid body (MNTB) synaptic transmission in the mammalian auditory brain stem mediate many excitatory transmitters such as glutamate, which is a useful model to study excitatory synaptic development. Hearing deficits are often accompanied by changes in the synaptic organization such as excitatory or inhibitory circuits as well as anatomical changes. Owing to this, circling mouse whose cochlea degenerates spontaneously after birth, is an excellent animal model to study deafness pathophysiology. However, little is known about the development regulation of the subunits composing these receptors in circling mouse. Thus, we used immunohistochemical method to compare the N-Methyl-D-aspartate receptor (NMDA receptor) NR1, NR2A, NR2B distribution in the LSO which project glutamergic excitatory input into the auditory brainstem, in circling mouse of postnatal (p) 7 and 16, which have spontaneous mutation in the inner ear, with wild-type mouse. The relative NMDAR1 immunoreactive density of the LSO in circling mouse p7 was $128.67\pm8.87$ in wild-type, $111.06\pm8.04$ in heterozygote, and $108.09\pm5.94$ in homozygote. The density of p16 circling mouse was $43.83\pm10.49$ in wild-type, $40\pm13.88$ in heterozygote, and $55.96\pm17.35$ in homozygote. The relative NMDAR2A immunoreactive density of LSO in circling mouse p7 was $97.97\pm9.71$ in wild-type, $102.87\pm9.30$ in heterozygote, and $106.85\pm5.79$ in homozygote. The density of LSO in p16 circling was $47.4\pm20.6$ in wild-type, $43.9\pm17.5$ in heterozygote, and $49.2\pm20.1$ in homozygote. The relative NMDAR2B immunoreactive density of LSO in circling mouse p7 was $109.04\pm6.77$ in wild-type, $106.43\pm10.24$ in heterozygote, and $105.98\pm4.10$ in homozygote. the density of LSO in p16 circling mouse was $101.47\pm11.5$ in wild-type, $91.47\pm14.81$ in heterozygote, and $93.93\pm15.71$ in homozygote. These results reveal alteration of NMDAR immunoreactivity in LSO of p7 and p16 circling mouse. The results of the present study are likely to be relevant to understand the central change underlying human hereditary deafness.

The Respiratory and Hemodynamic Effects of Prone Position According to the Level of PEEP in a Dog Acute Lung Injury Model (잡종견 급성폐손상 모델에서 Prone position 시행시 PEEP 수준에 따른 호흡 및 혈류역학적 효과)

  • Lim, Chae-Man;Chin, Jae-Yong;Koh, Youn-Suck;Shim, Tae-Sun;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.1
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    • pp.140-152
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    • 1998
  • Background: Prone position improves oxygenation in patients with ARDS probably by reducing shunt Reduction of shunt in prone position is thought to be effected by lowering of the critical opening pressure (COP) of the dorsal lung because the pleural pressure becomes less positive in prone position compared to supine position. It can then be assumed that prone position would bring about greater improvement in oxygenation when PEEP applied in supine position is just beneath COP than when PEEP is above COP. Hemodynamically, prone position is expected to attenuate the lifting of cardiac fossa induced by PEEP. Based on these backgrounds, we investigated whether the effect of prone position on oxygenation differs in magnitude according to the level of PEEP applied in supine position, and whether impaired cardiac output in supine position by PEEP can be restored in prone position. Methods: In seven mongrel dogs, $PaO_2/F_1O_2$(P/F) was measured in supine position and at prone position 30 min. Cardiac output (CO), stroke volume (SV), pulse rate (PR), and pulmonary artery occlusion pressure (PAOP) were measured in supine position, at prone position 5 min, and at prone position 30 min. After ARDS was established with warmed saline lavage(P/F ratio $134{\pm}72$ mm Hg), inflection point was measured by constant flow method($6.6{\pm}1.4cm$ $H_2O$), and the above variables were measured in supine and prone positions under the application of Low PEEP($5.0{\pm}1.2cm$ $H_2O$), and Optimal PEEP($9.0{\pm}1.2cm$ $H_2O$)(2 cm $H_2O$ below and above the inflection point, respectively) consecutively. Results : P/F ratio in supine position was $195{\pm}112$ mm Hg at Low PEEP and $466{\pm}63$ mm Hg at Optimal PEEP(p=0.003). Net increase of P/F ratio at prone position 30 min, however, was far greater at Low PEEP($205{\pm}90$ mm Hg) than at Optimal PEEP($33{\pm}33$ mm Hg)(p=0.009). Compared to CO in supine position at Optimal PEEP($2.4{\pm}0.5$ L/min), CO in prone improved to $3.4{\pm}0.6$ L/min at prone position 5 min (p=0.0180) and $3.6{\pm}0.7$ L/min at prone position 30 min (p=0.0180). Improvement in CO was attributable to the increase in SV: $14{\pm}2$ ml in supine position, $20{\pm}2$ ml at prone position 5 min (p=0.0180), and $21{\pm}2$ ml at prone position 30 min (p=0.0180), but not to change in PR or PAOP. When the dogs were turned to supine position again, MAP ($92{\pm}23$ mm Hg, p=0.009), CO ($2.4{\pm}0.5$ L/min, p=0.0277) and SV ($14{\pm}1$ ml, p=0.0277) were all decreased compared to prone position 30 min. Conclusion: Prone position in a dog with saline-lavaged acute lung injury appeared to augment the effect of relatively low PEEP on oxygenation, and also attenuate the adverse hemodynamic effect of relatively high PEEP. These findings suggest that a PEEP lower than Optimal PEEP can be adopted in prone position to achieve the goal of alveolar recruitment in ARDS avoiding the hemodynamic complications of a higher PEEP at the same time.

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