Abnormal Adhesion Research Articles

The multiple roles of macrophages in peritoneal adhesion

AbstractPeritoneal adhesion (PA) refers to the abnormal adhesion of the peritoneum either with the peritoneum itself or with tissues and organs that is caused by abdominopelvic surgery, abdominal infection or peritoneal inflammation. PA is associated with various clinical complications, such as abdominal pain and distension, intestinal obstruction, gastrointestinal disorders and female infertility, and adversely affects the quality of life of patients. Macrophages are essential for PA formation and can undergo polarization into classically activated macrophages (M1) and alternatively activated macrophages (M2), which are influenced by the peritoneal microenvironment. By releasing proinflammatory cytokines and reactive oxygen species, M1 macrophages promote peritoneal inflammatory reactions and the resultant formation of adhesion. In contrast, M2 macrophages secrete anti‐inflammatory cytokines and growth factors to inhibit PA formation and to promote repair and healing of peritoneal tissues, and thereby play a significant anti‐inflammatory role. This review comprehensively explores the function and mechanism of macrophages and their subtypes in PA formation to gain insight into the prevention and treatment of PA based on the modulation of macrophages.

Isoechinulin B, a natural product from Antarctic fungus, attenuates acute liver injury by inhibiting excessive cell adhesion

Abnormal cell adhesion between leukocytes and endothelial cells is closely associated with the development of numerous inflammation-related diseases, with adhesion molecules playing a crucial role. The disruption of cell adhesion directly or indirectly inhibits excessive cell adhesion and thus produces a therapeutic effect. However, there are only a few clinically available antagonists of cell adhesion. One of the biggest challenges is the development of novel and efficient cell adhesion inhibitors. Recently, the anti-inflammatory pharmacological activity of natural products of microbial origin has also received increasing attention. Here, we obtained a potential cell adhesion inhibitor isoechinulin B, an indole diketopiperazine derivative, from the Antarctic fungus Aspergillus sp. CPCC 401072, which is active against cell adhesion. Isoechinulin B decreased the expression of vascular endothelial adhesion molecule 1 (VCAM-1) and intercellular adhesion molecule 1 (ICAM-1) by inhibiting the activation of the NF-κB signaling pathway, thereby inhibiting cell adhesion between leukocytes and endothelial cells to reduce macrophage infiltration in the liver and significantly attenuate lipopolysaccharide-induced acute liver injury in mice. ConclusionIsoechinulin B is a novel cell adhesion inhibitor derived from fungi found in extreme environments.

Intestinal Obstruction Due to a Congenital Band: Two Cases of Congenital Bands in Adults

Small bowel obstruction is the most common surgical disorder of the small intestine. Adhesions are by far the most frequent causes followed by hernias, tumors, intussusception, foreign bodies, gallstones, and inflammatory bowel disease. During embryogenesis, abnormal adhesion of the peritoneal folds induces a congenital band which can cause small bowel obstruction. It is extremely rare and usually observed in childhood. We report two cases of small bowel obstructions in adults due to congenital bands. The first patient is a woman of 47 years-old admitted for chronic abdominal pain, and the second one is a 20 years-old man admitted for the etiological assessment of chronic constipation. Surgical exploration was performed revealing a small bowel obstruction due to congenital band with complete resolution of the symptoms after surgery.

Examination of Subbasal Nerve Plexus and Central Corneal Stromal Microstructure in Subjects With Congenital Aniridia, Using in Vivo Confocal Laser Scanning Microscopy

Purpose During life up to 70% of aniridia subjects develop aniridia-associated keratopathy (AAK). AAK is characterized by limbal stem cell insufficiency, impaired corneal epithelial cell differentiation and abnormal cell adhesion, which leads to centripetal spreading vascularization, conjunctivalization, and thickening of the cornea. Our aim was to examine the subbasal nerve plexus and central corneal stromal microstructure in subjects with congenital aniridia, using in vivo confocal laser scanning microscopy CLSM. Methods 31 eyes of 18 patients (55.6% males, mean age: 25.22 ± 16.35 years) with congenital aniridia and 46 eyes of 29 healthy subjects (41.4% males, mean age 30 ± 14.82 years) were examined using the Rostock Cornea Module of Heidelberg Retina Tomograph-III. At the subbasal nerve plexus, corneal nerve fiber density (CNFD), corneal nerve fiber length (CNFL), corneal total branch density (CTBD), and corneal nerve fiber width (CNFW) were analyzed using ACCMetrics software. Keratocyte density in the anterior, middle and posterior stroma was assessed manually. Results The CNFD (2.02 ± 4.08 vs 13.99 ± 6.34/mm2), CNFL (5.78 ± 2.68 vs 10.56 ± 2.82 mm/mm2) and CTBD (15.08 ± 15.62 vs 27.44 ± 15.05/mm2) were significantly lower in congenital aniridia subjects than in controls (p < 0.001 for all). CNFW was significantly higher in aniridia subjects than in controls (0.03 ± 0.004 vs 0.02 ± 0.003 mm/mm2) (p = 0.003). Keratocyte density was significantly lower in all stromal layers of aniridia subjects than in controls (p < 0.001 for all). Stromal alterations included confluent keratocytes, keratocytes with long extensions and hyperreflective dots between keratocytes in aniridia. Conclusions Decrease in CNFD, CNFL, and CTBD, as well as increase in CNFW well refer to the congenital aniridia-associated neuropathy. The decreased keratocyte density and the stromal alterations may be related to an increased cell death in congenital aniridia, nevertheless, stromal changes in different stages of AAK have to be further analyzed in detail.

The sub‐basal nerve plexus and central corneal stromal microstructure in subjects with congenital aniridia, using in vivo confocal microscopy

Purpose: Congenital aniridia is a rare disease, with a global prevalence of 1 in 40 000 to 1 in 100 000. During life up to 70% of aniridia sufferers develop aniridia‐associated keratopathy (AAK) with limbal stem cell insufficiency, impaired corneal epithelial cell differentiation, abnormal cell adhesion and wound healing. AAK is characterized by centripetal spreading vascularization, conjunctivalization, and thickening of the cornea. We examined the sub‐basal nerve plexus and central corneal stromal microstructure in subjects with congenital aniridia, using in vivo confocal laser scanning microscopy (IVCM).Methods: 31 eyes of 18 patients (55.6% males, mean age: 25.22 ± 16.35 years) with congenital aniridia and 47 eyes of 29 healthy subjects (41.4% males, mean age 30 ± 14.82 years) were examined using the Rostock Cornea Module of Heidelberg Retina Tomograph‐III. At the sub‐basal nerve plexus, corneal nerve fibre density (CNFD), corneal nerve fibre length (CNFL), corneal total branch density (CTBD) and corneal nerve fibre width (CNFW) were analysed using ACCMetrics software. Keratocyte density in the anterior, middle and posterior stroma was assessed manually.Results: The CNFD (2.02 ± 4.08 vs. 13.99 ± 6.34/mm2), CNFL (5.78 ± 2.68 vs. 10.56 ± 2.82 mm/mm2) and CTBD (15.08 ± 15.62 vs. 27.44 ± 15.05/mm2) were significantly lower in congenital aniridia subjects, than in controls (p &lt; 0.001 for all). CNFW was significantly higher in aniridia subjects, than in controls (0.03 ± 0.004 vs. 0.02 ± 0.003 mm/mm2) (p &lt; 0.003). Keratocyte density was significantly lower in all stromal layers of congenital aniridia subjects, than in controls (p &lt; 0.001 for all). Stromal alterations included confluent keratocytes, keratocytes with long extensions and hyperreflective dots between keratocytes in congenital aniridia.Conclusions: CNFD, CNFL and CTBD decrease and CNFW increase well refer to the congenital aniridia associated neuropathy. The decreased keratocyte density contradicts the inflammatory hypothesis of congenital aniridia. Corneal stromal changes in different stages of AAK have to be further analysed in detail0.

Adhesion Reduction Agent Guardix-SG® Versus MegaShield® for Postoperative Swallowing Function Analysis in Thyroidectomy Patients.

Antiadhesion products are essential for postoperative care in patients after thyroidectomy by providing a physical barrier to cover the exposed tissue and thus preventing abnormal adhesion of adjacent tissues. Since thyroidectomy may result in swallowing difficulties arising from damage or inflammation of the surrounding tissues, the use of antiadhesion agents such as MegaShield® or Guardix-SG® will help reduce scar formation. This may thus improve postoperative swallowing function in patients. Patients were enrolled and followed up between October 4, 2018, and March 26, 2020. Patients during the postoperative follow-up sessions were randomly allocated to the standard care with Guardix-SG® and clinical trial medical device application group with MegaShield® (test group) in a 1:1 ratio by the permuted block randomization method. Patient performance on penetration aspiration scale (PAS), National Institutes of Health-Swallow Safety Scale (NIH-SSS), videofluoroscopic dysphagia scale (VDS), Dynamic Imaging Grade of Swallowing Toxicity (DIGEST) based on Videofluoroscopic swallowing study (VFSS) were collected. Nonadhesion-reducing agent patient data were used as a control group. No statistical significance was shown (P > .05) between the 2 groups of MegaShield® and Guardix-SG® in various phases from thick semisolid, thin semisolid to liquid for both PAS and NIH-SSS. Several statistical significances were reported in the results comparing various criteria of PAS, NIH-SSS, VDS at different oral and pharyngeal phases, and DIGEST in all 3 stages among MegaShield®, Guardix-SG®, and nonadhesion-reducing agent group. These results prove the noninferiority of MegaShield® compared with Guardix-SG® as an antiadhesion agent in postthyroidectomy care.

Placenta accreta on non-scarred uterus: About a case

Placenta accreta is an abnormally adherent placenta (abnormal adhesion of the placenta to the myometrium), due to the localized or diffuse absence of the decidua basalis. Its incidence increases with increasing cesarean section rates. Placenta accreta in a non-scarred uterus is very rare but remains possible due to other risk factors which are age, smoking, placenta preavia and finally invasive endouterine procedures. Among its consequences, we first note the hemorrhage of delivery which can be associated with significant maternal morbidity and mortality, uterine rupture and also the invasion of adjacent organs by the placental trophoblast. We report an interesting case of placenta accreta on a non-scarred uterus discovered during a planned cesarean section for placenta previa. This is a 41-year-old patient with no history of scarred uterus, 5th procedure, 3rd parity, with history of spontaneous miscarriage cured at 2 months, pregnant at 39 weeks, unmonitored pregnancy, admitted to the maternity emergency room for two days before for delivery method where the parturient benefited from a clinical examination and an obstetric ultrasound showing a completely covering anterior placenta, amniotic fluid of normal quantity and an estimate of the fetal weight at 2800g, then scheduled for a cesarean section. The diagnosis of placenta accreta is made during cesarean section. We opted for radical treatment. Placenta accreta is a pathology at risk of serious hemorrhagic complications during pregnancy and the postpartum period. This should encourage us to systematically search for ultrasound criteria for placenta accreta. The ultrasound report, in these patients, must explicitly mention this. Adequate care in the presence of a multidisciplinary team makes it possible to limit the mortality and morbidity associated with this pathology.

Radchip: A Laminin-Lined Red Blood Cell (RBC) Adhesion Device for Functional Characterization and Clinical Evaluation of Sickle RBCs at Steady-State

Sickle cell disease (SCD) is a genetic disorder caused by a point mutation in the beta-globin gene producing an abnormal hemoglobin (HbS) which, under deoxygenation, polymerizes and causes severe distortion of the red blood cell (RBC). RBCs may be dehydrated, poorly deformable and highly adhesive to the endothelium, with abnormal adhesion being of primary interest to this study. Recently, several new disease modifying drugs have been FDA approved, with more in development. These novel agents target specific aspects of SCD pathophysiology, including adhesion. Changes induced by such therapies are not always detectable by common conventional lab instruments. LORRCA (oxygen-gradient ektacytometry) is the gold-standard for deformability measurement; it has been well described both analytically and clinically in SCD. Adhesion and deformability have a dynamic, yet complementary relationship that we explore here. While these may behave in an interdependent manner, each determinant must be considered independently. There is a growing demand for innovative tools to evaluate rheological properties under physiological conditions, particularly microfluidic devices. However, many current devices are limited in their clinical utility due to, choice of ligand, complexity of design, or reliance on endothelial cell culture (typically HUVEC cells) which are difficult to use, vary with passage number, and do not represent adult SCD endothelium. We developed a Red blood cell Adhesion Chip (RAdChip) to model RBC to endothelial adhesion using laminin, a physiologically relevant ligand, to easily, effectively, and reproducibly measure RBC adhesion. Here, we describe our analytic and clinical validation of our device. Peripheral blood samples were collected in EDTA from individuals with SCD and healthy controls under an IRB approved protocol. RBCs were extracted via centrifugation, washed and resuspended in PBS to a 0.5% HCT. Microfluidic devices were fabricated using polydimethylsiloxane (PDMS). A 5ug/ml solution of recombinant laminin was applied to each channel, then blocked with 1% bovine serum albumin (BSA). Resuspended RBCs were introduced to the device through tubing connected to the inlet port. Harvard Apparatus syringe pumps were used to perfuse samples and wash channels at ~1dyne/cm 2. Adhesion was visualized with a Keyence inverted bright-field microscope at 20x (Itasca, IL). Adhered cells were counted within a 1.8mm 2 region of interest (Roi) using the iCLOTS automated cell quantification software (Fay et al, Nat comm accepted 7/2023). We measured adhesion index via RAdChip for common SCD genotypes: HbSS/SB0: (median (x͂) =756; range (r): 154-1889; n=26) HbSC/SB+ : (x͂ =299; r=: 114-731; n=11 ) HbAA: (x͂) =13; r= 5-90; n=7). RBC adhesion values were independent of age (p=0.65) and gender (p=0.63). An increase of 0.1 unit in EImin and EImax decreases RBC adhesion by 214 and 268 cells/mm 2, respectively. A 1mmHg increase in PoS increased adhesion by 19.5 cells/mm 2. sRBC adhesion had a significant positive association with vaso-occlusive events (VOE) (p=0.005), absolute neutrophil count (ANC) (p=0.005), and EImax (p&amp;lt;0.001). Notably, analytical validation of RAdChip produced a mean CV of 0.13 ± 0.94. RAdChip adds a key component of functional RBC evaluation that conventional lab equipment does not measure. We have analytically validated our device, demonstrating an excellent coefficient of variability. 44 patient samples, each examined in triplicate produced a mean CV of 0.13. We have demonstrated clinical validity of our device by establishing correlations with known markers of severity and pain event frequency. RBC adhesion values increase as the number of VOE events increase, reaffirming the belief that adhesion is a principal element in the multistep VOE cascade. In like manner, improvements in deformability are associated with decreased RBC adhesion, strengthening the interdependent nature of VOE molecular events. Recognizing the dynamic nature of SCD, it is necessary to explore sickle erythrocyte adhesion longitudinally to investigate changes before, during, and after VOE occurrences. Continued and increased integration of adhesion measurements with RAdChip and other rheology devices into routine SCD care may be helpful in assessing disease severity, identifying need for additional disease modifying agents, and assist with second line agent selection and monitoring.

Impact of Maternal Preoperative Hemoglobin Levels on Intraoperative Hemorrhage Risk in Placenta Accrete Spectrum Disorders: A Retrospective Cross-Sectional Study.

BACKGROUND Placenta accreta spectrum (PAS) disorders involve abnormal adhesion or invasion of chorionic villi through the myometrium and uterine serosa. Maternal anemia during pregnancy is common and may contribute to complications during delivery, particularly with abnormal placentation. This study examines the association between preoperative maternal hemoglobin levels and the risk of intraoperative massive hemorrhage in pregnant women with PAS disorders. MATERIAL AND METHODS A retrospective study included 538 consecutive participants (mean age=31.12&plusmn;4.68 years) who underwent cesarean sections and met the diagnostic criteria for PAS disorders. Logistic regression analysis was performed to investigate the relationship between maternal preoperative hemoglobin levels and the risk of massive intraoperative hemorrhage (blood loss ≥1500 mL). RESULTS The incidence of intraoperative massive hemorrhage among patients with PAS disorders was 38.66%. The mean preoperative maternal hemoglobin level was 10.99&plusmn;1.39 g/dL, and overall anemia incidence (<11 g/dL) was 48.88% in our study. After adjusting for potential confounders, a non-linear relationship was observed between preoperative maternal hemoglobin levels and the risk of intraoperative massive hemorrhage. When the preoperative hemoglobin level of pregnant women was below 11.5 g/dL (OR=0.52, 95% CI 0.39-0.70), the lower hemoglobin level significantly increased the risk of intraoperative hemorrhage. CONCLUSIONS Maternal preoperative hemoglobin levels were inversely associated with the risk of massive intraoperative hemorrhage in PAS disorders. A non-linear relationship was identified, with a turning point at 11.5 g/dL. These findings emphasize the importance of monitoring and managing maternal hemoglobin levels to mitigate the risk of intraoperative hemorrhage in pregnant women with PAS disorders.

Chronic hyperglycemia reduces the expression of intercellular adhesion molecules and increases intercellular hyperpermeability in the periodontal epithelium.

Hyperglycemia in diabetes is closely associated with periodontal disease progression. This study aimed to investigate the effect of hyperglycemia on the barrier function of gingival epithelial cells as a cause of hyperglycemia-exacerbated periodontitis in diabetes mellitus. The abnormal expression of adhesion molecules in gingival epithelium in diabetes was compared between db/db and control mice. To study the effects of hyperglycemia on interepithelial cell permeability, the mRNA and protein expressions of adhesion molecules were investigated using a human gingival epithelial cell line (epi 4 cells) in the presence of either 5.5 mM glucose (NG) or 30 mM glucose (HG). Immunocytochemical and histological analyses were performed. We also studied HG-related intracellular signaling to assess abnormal adhesion molecule expression in the cultured epi 4 cells. The results of the proteomic analysis implied the abnormal regulation of cell-cell adhesion, and mRNA and protein expression assessments revealed the significant downregulation of Claudin1 expression in the gingival tissues of db/db mice (p < .05 vs control). Similarly, the mRNA and protein expressions of adhesion molecules were lower in epi 4 cells cultured under HG conditions than in those cultured under NG conditions (p < .05). Three-dimensional culture and transmission electron microscopy revealed reduced thickness of the epithelial cell layers with no flattened apical cells and heterogeneously arranged intercellular spaces among adjacent epi 4 cells under the HG. These results were consistent with the increased permeability of epi 4 cells under the HG relative to that of cells under the NG. This abnormal expression of intercellular adhesion molecules under the HG was related to the increased expression of receptors for advanced glycation end products (AGEs) and oxidative stress relative to that seen under the NG, along with stimulation of ERK1/2 phosphorylation in epi 4 cells. High glucose-induced impairment of intercellular adhesion molecule expression in gingival epithelial cells was related to the intercellular permeability of gingival cells, representing a possible link to hyperglycemia-related AGE signaling, oxidative stress, and ERK1/2 activation.

Placenta accreta spectrum with severe morbidity: fibrosis associated with cervical-trigonal invasion

Objective Describe the clinical-surgical results of patients with PAS in the low-posterior cervical-trigonal space associated with fibrosis (PAS type 4) compared with PAS types in other locations (Types 1, upper bladder, 2 in upper parametrium) and in particular with PAS type 3, corresponding to dissectible cervical-trigonal invasion. The clinical-surgical results of using a standard hysterectomy were analyzed with a modified subtotal hysterectomy (MSTH) in patients with PAS type 4. Material and methods A descriptive, retrospective, multicenter study included 337 patients of PAS; thirty-two corresponding to PAS type 4, from three PAS reference hospitals, CEMIC, Buenos Aires, Argentina, Fundación Valle de Lili, Cali, Colombia, and Dr. Soetomo General Hospital, Surabaya, Indonesia, between January 2015 and December 2020. PAS was diagnosed by abdominal and transvaginal ultrasound and topographically characterized by ultrafast T2 weighted MRI. In persistent macroscopic hematuria after MSTH, the surgeon performs an intentional cystotomy and uses a square compression suture to achieve the hemostasis inside the bladder wall. According to a PAS topographical classification, the patients with low-vesical cervical involvement compared with PAS located in relation with the upper blader (type1), upper parametrium (type 2 upper), and also with PAS situated in the lower vesical-trigon space (type 3). PAS 3 and 4 are located in identical area, but in type 3, group A, the vesicouterine space was dissectible, and in type 4, group B, significant fibrosis made surgical dissection extremely challenging. Furthermore, group B was divided into patients treated with total hysterectomy (HT) and those treated with a modified subtotal hysterectomy (MSTH). The surgical requirements to perform an MSHT included the availability of proximal vascular control at the aortic level (internal manual aortic compression, aortic endovascular balloon, aortic loop, or aortic cross-clamping). Then surgeon performed an upper segmental hysterotomy, avoiding the abnormal placenta invasion area; after that, the fetus was delivered, and the umbilical cord was ligated. After uterine exteriorization, the surgeon applies a continuous circular suture with number 2 polyglactin 910, taking some portions of the myometrium -to avoid unintentional slipping- around the lower uterine segment and a 3–4 cm proximal to the abnormal adhesion of the placenta. After tightening hard the circular suture, the uterine segment was circumferentially cut, three centimeters proximal to the circular hemostatic sutures. Next, the surgery follows the upper steps of conventional hysterectomy without changes. Additionally, the histological presence of fibrosis was examined in all samples. Results Modified subtotal hysterectomy in patients with PAS type 4 (cervical-trigonal fibrosis) resulted in a significant clínico-surgical improvement over total hysterectomy. The median operative time and intraoperative bleeding were 140 min (IQR 90–-240) and 1895 mL (IQR 1300–2500) in patients undergoing modified subtotal hysterectomy, and 260 min (IQR 210–287) and 2900 mL (IQR 2150–5500) in patients treated with total hysterectomy, respectively. The complication rate was 20% for MSHT and 82.3% for patients with a total hysterectomy. Conclusions PAS in the cervical trigonal area associated with fibrosis implies a greater risk of complications due to uncontrollable bleeding and organ damage. MSTH is associated with lower morbidity and difficulties in PAS type 4. Prenatal or intrasurgical diagnosis is essential to plan surgical alternatives to improve the results.

Membrane bending and sphingomyelinase-associated, sulfatide-dependent hypoxic adhesion of sickle mature erythrocytes.

Abnormal erythrocyte adhesion owing to polymerization of sickle hemoglobin is central to the pathophysiology of sickle cell disease (SCD). Mature erythrocytes constitute >80% of all erythrocytes in SCD; however, the relative contributions of erythrocytes to acute and chronic vasculopathy in SCD are not well understood. Here, we showed that bending stress exerted on the erythrocyte plasma membrane by polymerization of sickle hemoglobin under hypoxia, enhances sulfatide-mediated abnormal mature erythrocyte adhesion. We hypothesized that sphingomyelinase (SMase) activity, which is upregulated by accumulated bending energy, leads to elevated membrane sulfatide availability, and thus, hypoxic mature erythrocyte adhesion. We found that mature erythrocyte adhesion to laminin in controlled microfluidic experiments is significantly greater under hypoxia than under normoxia (1856± 481 vs 78± 23, mean± SEM), whereas sickle reticulocyte (early erythrocyte) adhesion, high to begin with, does not change (1281± 299 vs 1258± 328, mean± SEM). We showed that greater mean accumulated bending energy of adhered mature erythrocytes was associated with higher acid SMase activity and increased mature erythrocyte adhesion (P= .022, for acid SMase activity and P= .002 for the increase in mature erythrocyte adhesion with hypoxia, N= 5). In addition, hypoxia results in sulfatide exposure of the erythrocyte membrane, and an increase in SMase, whereas anti-sulfatide inhibits enhanced adhesion of erythrocytes. These results suggest that the lipid components of the plasma membrane contribute to SCD complications. Therefore, sulfatide and the components of its upregulation pathway, particularly SMase, should be further explored as potential therapeutic targets for inhibiting sickle erythrocyte adhesion.

CASK silencing favors systolic function by regulating junction's signaling

Although the organization of the cardiomyocyte (CM) membrane into functional microdomains directly influences the anisotropic propagation of the AP, the mechanisms underlying the rod-like structural organization of CMs during post-natal development are poorly understood. We have previously shown in the adult CM that CASK regulates sodium channel expression at the lateral membrane in interaction with focal adhesion complexes. In addition, preliminary results have shown that the expression level of CASK evolves during postnatal development and is altered during post-infarction myocardial remodeling in rat. To unravel the implication of CASK in postnatal cardiomyocyte differentiation and in the acquisition of structural and electrical function. CASK viral constructs were designed to inhibit the expression of CASK in neonatal cardiomyocytes, both in vitro (Ad-shCASK) and in vivo (AAV-shCASK). Western blots on human heart samples of arrythmogenic cardiomyopathy, dilated cardiomyopathy and atrial fibrilation patients revealed strong decrease of CASK expression. Proteomic analysis showed that CASK invalidation strongly modify immature CM proteome, notably junction signaling. Immunostaining experiments on neonatal CMs confirmed that CASK is required for the correct organization of the cytoskeleton and differentially regulates both mechanical and electrical junction formation of the intercalated disc. Neonatal CMs invalidated for CASK displayed abnormal contractility and cell adhesion fragility. Stiffness measurements using SICM shows increased stiffness at the junction site, validating the modification of the cell-cell contacts’ physical properties. Echocardiography phenotyping of CASK-invalidated rats at the neonatal stage reveals eccentric ventricular remodeling, decreased systolic performance but a preserved contractile reserve. Pressure-volume measurements confirmed that CASK invalidation increased myocardium contractility and compliance. In addition, CASK silencing does not trigger electrical remodeling or spontaneous arrhythmias, despite the reduced Nav1.5/Kir2.1 expression level. This study demonstrates that CASK confers the ability to orchestrate both CM molecular organization and tissue cohesion. As the decrease of CASK expression appears to be a marker of cardiac remodeling, CASK overexpression therapy could be promising regarding hereditary cardiomyopathies.

Septin11 promotes hepatocellular carcinoma cell motility by activating RhoA to regulate cytoskeleton and cell adhesion

Septins as GTPases in the cytoskeleton, are linked to a broad spectrum of cellular functions, including cell migration and the progression of hepatocellular carcinoma (HCC). However, roles of SEPT11, the new member of septin, have been hardly understood in HCC. In the study, the clinical significance and biological function of SEPT11 in HCC was explored. SEPT11 was screened out by combining ATAC-seq with mRNA-seq. Role of SEPT11 in HCC was further investigated by using overexpression, shRNA and CRISPR/Cas9-mediated SEPT11-knockout cells or in vivo models. We found RNA-seq and ATAC-seq highlights LncRNA AY927503 (AY) induced SEPT11 transcription, resulting in Rho GTPase activation and cytoskeleton actin aggregation. The GTP-binding protein SEPT11 is thus considered, as a downstream factor of AY, highly expressed in various tumors, including HCC, and associated with poor prognosis of the patients. In vitro, SEPT11 overexpression promotes the migration and invasion of HCC cells, while SEPT11-knockout inhibits migration and invasion. In vivo, SEPT11-overexpressed HCC cells show high metastasis incidents but don’t significantly affect proliferation. Meanwhile, we found SEPT11 targets RhoA, thereby regulating cytoskeleton rearrangement and abnormal cell adhesion through ROCK1/cofilin and FAK/paxillin signaling pathways, promoting invasion and migration of HCC. Further, we found SEPT11 facilitates the binding of GEF-H1 to RhoA, which enhances the activity of RhoA. Overall, our study confirmed function of SEPT11 in promoting metastasis in HCC, and preliminarily explored its related molecular mechanism. SEPT11 acts as an oncogene in HCC, also draws further interest regarding its clinical application as a potential therapeutic target.

Prophylactic bilateral internal iliac balloon occlusion to control hemorrhage in placenta accreta spectrum: a boon for obstetricians.

Placenta accreta spectrum (PAS) is abnormal placental adhesion beyond superficial myometrium, which may lead to severe life-threatening hemorrhage requiring massive blood transfusions in the peripartum period. Prophylactic balloon catheterization of bilateral internal iliac arteries with or without additional embolization with Cesarean hysterectomy in patients with PAS prevent excessive intraoperative hemorrhage and may also obviate the need for hysterectomy. To study the efficacy of intervention radiological procedures in controlling intraoperative hemorrhage in patients with PAS. This ethically approved prospective study was conducted between November 2017 and October 2019 and written informed consent was obtained from all patients. Consecutive patients diagnosed with PAS during the antepartum period were evaluated. A total of 18 patients with PAS underwent prophylactic balloon catheterization of the bilateral internal iliac arteries followed by delivery of the infant. Interventional and intraoperative data of these patients were collected and compared with retrospectively collected data of patients (control group) who underwent hysterectomy without prophylactic balloon occlusion over the past four years (January 2016-November 2019). Significantly lower intraoperative blood loss (2.8 L vs. 4.7 L; P = 0.048) and pure red blood cell (PRBC) requirement (P = 0.026) between patients who had hysterectomy with and without interventional radiological management was observed. Significantly higher blood loss (P = 0.006) and fluid requirement (P = 0.007) was observed with a higher degree of placental invasion. Only 1 (6%) major procedure-related complication was observed. Interventional radiological procedures are effective in significantly reducing intraoperative blood loss and blood product requirement in patients with PAS.

Anatomic and Magnetic Resonance Imaging-Based Correction of Upper-Eyelid Depression and Blepharoptosis in Senile Patients.

In senile patients with sunken superior sulcus, involutional ptosis, and higher eyelid crease, a single operation to correct depression or ptosis cannot achieve good results. We demonstrated the anatomy of periorbital septum fibers, which may contribute to the levator muscle's volume depletion and dynamic power transmission disorder, and described a procedure for correcting upper-eyelid depression and blepharoptosis in senile patients. The fibrous webs in these patients connected the posterior aspect of the orbicularis and the orbital septum and extended to the orbital fat and levator aponeurosis. These fibers were dissected to release the periorbital septal fibers, and the orbital septal fat flap was transferred to the depressed region. Advancement or plication of the levator aponeurosis was performed in patients with uncorrected blepharoptosis after the procedures described above. The technique was applied to 13 Chinese patients (25 eyes) between May 2021 and April 2022. Postoperative magnetic resonance imaging revealed that the preaponeurotic fat was displaced forward and down to the upper margin of the tarsus, and the curvature of the upper-eyelid depression was significantly improved. Moreover, the superior sulcus deformity improved, the ptosis was corrected, and the uppermost crease decreased in all patients. No recurrence of ptosis or abnormal adhesion was observed. We believe this is the first study using magnetic resonance imaging to evaluate eyelid anatomy and the effects of surgery in this patient group. Releasing periorbital septum fibers is crucial for correcting a portion of the sunken eyelid and ptosis in Asians.

Sickle red blood cell-derived extracellular vesicles activate endothelial cells and enhance sickle red cell adhesion mediated by von Willebrand factor.

Endothelial activation and sickle red blood cell (RBC) adhesion are central to the pathogenesis of sickle cell disease (SCD). Quantitatively, RBC-derived extracellular vesicles (REVs) are more abundant from SS RBCs compared with healthy RBCs (AA RBCs). Sickle RBC-derived REVs (SS REVs) are known to promote endothelial cell (EC) activation through cell signalling and transcriptional regulation at longer terms. However, the SS REV-mediated short-term non-transcriptional response of EC is unclear. Here, we examined the impact of SS REVs on acute microvascular EC activation and RBC adhesion at 2 h. Compared with AA REVs, SS REVs promoted human pulmonary microvascular ECs (HPMEC) activation indicated by increased von Willebrand factor (VWF) expression. Under microfluidic conditions, we found abnormal SS RBC adhesion to HPMECs exposed to SS REVs. This enhanced SS RBC adhesion was reduced by haeme binding protein haemopexin or VWF cleaving protease ADAMTS13 to a level similar to HPMECs treated with AA REVs. Consistent with these observations, haemin- or SS REV-induced microvascular stasis in SS mice with implanted dorsal skin-fold chambers that was inhibited by ADAMTS13. The adhesion induced by SS REVs was variable and was higher with SS RBCs from patients with increased markers of haemolysis (lactate dehydrogenase and reticulocyte count) or a concomitant clinical diagnosis of deep vein thrombosis. Our results emphasise the critical contribution made by REVs to the pathophysiology of SCD by triggering acute microvascular EC activation and abnormal RBC adhesion. These findings may help to better understand acute pathophysiological mechanism of SCD and thereby the development of new treatment strategies using VWF as a potential target.

Prediction of Placenta Accreta Spectrum by Machine Learning Methods and Determination of Candidate Biomarkers

Placenta accreta spectrum (PAS) disorders; Abnormal adhesion of placental villi to the myometrium associated with endometrial trauma or dysplasia. Placenta previa and previous cesarean section operations are two major risk factors for PAS disorders. It is usually diagnosed by ultrasound examinations performed during pregnancy follow-up. After this diagnosis is made, a very careful and strict pregnancy follow-up should be done. If the diagnosis is made during pregnancy, the delivery should be done by cesarean section and the bleeding that the mother will experience should be stopped with an appropriate method. However, no protein candidate to be used in clinical diagnosis has been found so far. The aim of this study is to identify candidate biomarkers that can be used in the diagnosis and follow-up of PAS with machine learning methods.&#x0D; In this study, proteomic data obtained from 26 women with and without PAS were used. After using the Lasso method as the variable selection method, machine learning models (XGBoost, Adaboost) were created with 5-fold cross-validation. Accuracy, Balanced accuracy, Sensitivity, Specificity, Positive Predictive Value, Negative Predictive Value, F1-Score, MCC and G-mean metrics were used in the performance evaluation of the models created.&#x0D; When the performance metrics of the two models are compared, the best result belongs to the XGBoost machine learning model. Therefore, the Accuracy, Balanced accuracy, Sensitivity, Specificity, Positive Predictive Value, Negative Predictive Value, F1-Score, MCC, and G-mean performance criteria for the XGBoost model are 0.962, 0.950, 1.00, 0.90, 0.94, 1.00, 0.97, 0.92, and 0.97, respectively.&#x0D; As a result, considering the experimental results, it can be said that the created machine learning model is quite successful in classifying PAS. In addition, it can be said that KDR and AMH proteins are candidate biomarkers that can be used in the diagnosis and follow-up of PAS according to the significance of the variables related to the model.

Honeycomb-like Structured Film, a Novel Therapeutic Device, Suppresses Tumor Growth in an In Vivo Ovarian Cancer Model.

Ovarian cancer cell dissemination can lead to the mortality of patients with advanced ovarian cancer. Complete surgery for no gross residual disease contributes to a more favorable prognosis than that of patients with residual disease. HCFs have highly regular porous structures and their 3D porous structures act as scaffolds for cell adhesion. HCFs are fabricated from biodegradable polymers and have been widely used in tissue engineering. This study aimed to show that HCFs suppress tumor growth in an in vivo ovarian cancer model. The HCF pore sizes had a significant influence on tumor growth inhibition, and HCFs induced morphological changes that rounded out ovarian cancer cells. Furthermore, we identified gene ontology (GO) terms and clusters of genes downregulated by HCFs. qPCR analysis demonstrated that a honeycomb structure downregulated the expression of CXCL2, FOXC1, MMP14, and SNAI2, which are involved in cell proliferation, migration, invasion, angiogenesis, focal adhesion, extracellular matrix (ECM), and epithelial-mesenchymal transition (EMT). Collectively, HCFs induced abnormal focal adhesion and cell morphological changes, subsequently inhibiting the differentiation, proliferation and motility of ovarian cancer cells. Our data suggest that HCFs could be a novel device for inhibiting residual tumor growth after surgery, and could reduce surgical invasiveness and improve the prognosis for patients with advanced ovarian cancer.

Perfluorocarbon liquid-assisted vitreo-dissection in eyes with firmly adherent posterior hyaloid

BackgroundInduction of posterior vitreous detachment (PVD) is a critical step during pars plana vitrectomy. Multiple techniques and utilities have been proposed for assistance with this step with no consensus on the safest and most effective means, especially in eyes with firmly adherent posterior hyaloid. Viscodissection or the utilization of perfluorocarbon liquid (PFCL) can be used to dissect the posterior hyaloid and widely adherent epiretinal membranes.MethodsA technique of PFCL dissection of the posterior hyaloid in eyes with abnormal adhesion of the posterior hyaloid. After core vitrectomy, breaking into the posterior hyaloid face is made via active aspiration and cutting or a sharp dissection. This is followed by active and slow injection of PFCL into the potential space between the posterior cortical vitreous and the neurosensory retina. A wave of PFCL propagates anteriorly causing “vitreo-dissection” of the peripheral cortical vitreous.ResultsThe technique was effective and safe in 8 successive cases, 4 cases with vitreoretinal traction syndrome and 4 with diabetic tractional membranes.ConclusionThe technique can be considered in cases with abnormal firmly adherent posterior hyaloid when induction of PVD proves difficult.