Rap1 Small GTPase Regulates Vascular Endothelial-Cadherin-Mediated Endothelial Cell-Cell Junctions and Vascular Permeability.

The vascular permeability of the endothelium is finely controlled by vascular endothelial (VE)-cadherin-mediated endothelial cell-cell junctions. In the majority of normal adult tissues, endothelial cells in blood vessels maintain vascular permeability at a relatively low level, while in response to inflammation, they limit vascular barrier function to induce plasma leakage and extravasation of immune cells as a defense mechanism. Thus, the dynamic but also simultaneously tight regulation of vascular permeability by endothelial cells is responsible for maintaining homeostasis and, as such, impairments of its underlying mechanisms result in hyperpermeability, leading to the development and progression of various diseases including coronavirus disease 2019 (COVID-19), a newly emerging infectious disease. Recently, increasing numbers of studies have been unveiling the important role of Rap1, a small guanosine 5'-triphosphatase (GTPase) belonging to the Ras superfamily, in the regulation of vascular permeability. Rap1 enhances VE-cadherin-mediated endothelial cell-cell junctions to potentiate vascular barrier functions via dynamic reorganization of the actin cytoskeleton. Importantly, Rap1 signaling activation reportedly improves vascular barrier function in animal models of various diseases associated with vascular hyperpermeability, suggesting that Rap1 might be an ideal target for drugs intended to prevent vascular barrier dysfunction. Here, we describe recent progress in understanding the mechanisms by which Rap1 potentiates VE-cadherin-mediated endothelial cell-cell adhesions and vascular barrier function. We also discuss how alterations in Rap1 signaling are related to vascular barrier dysfunction in diseases such as acute pulmonary injury and malignancies. In addition, we examine the possibility of Rap1 signaling as a target of drugs for treating diseases associated with vascular hyperpermeability.

[Retrospective analysis of nosocomial COVID-19: a comparison between patients with hematological disorders and other diseases].

Nosocomial coronavirus disease 2019 (COVID-19) had occurred at our hospital. We retrospectively analyzed the differences between patients with nosocomial COVID-19 and either hematological disease (n=40) or other diseases (n=57). The analysis was completed within 60 days for surviving patients. Among the patients with hematological disease and those with other diseases, there were 21 (52.5%) and 20 (35.1%) deaths, respectively. Although the patients with hematological disease received favipiravir more frequently than patients with other diseases (21 [52.5%] vs. 15 [35.3%], respectively; P<0.05), their median overall survival was poor (29 days; P=0.078). Furthermore, the median duration from oxygen therapy initiation to death or intubation was significantly shorter in the patients with hematological disease (5 days [range, 1-17 days] vs. 10 days [1-24 days], respectively; P<0.05). Furthermore, the patients with hematological disease and nosocomial COVID-19 exhibited more marked respiratory failure and poorer outcomes leading to death in a shorter time period than the patients with other diseases and nosocomial COVID-19.

3-Hz subthreshold oscillations of CA2 neurons In vivo.

The CA2 region is unique in the hippocampus; it receives direct synaptic innervations from several hypothalamic nuclei and expresses various receptors of neuromodulators, including adenosine, vasopressin, and oxytocin. Furthermore, the CA2 region may have distinct brain functions, such as the control of instinctive and social behaviors; however, little is known about the dynamics of the subthreshold membrane potentials of CA2 neurons in vivo. We conducted whole-cell current-clamp recordings from CA2 pyramidal cells in urethane-anesthetized mice and monitored the intrinsic fluctuations in their membrane potentials. The CA2 pyramidal cells emitted spontaneous action potentials at mean firing rates of ∼0.8 Hz. In approximately half of the neurons, the subthreshold membrane potential oscillated at ∼3 Hz. In two neurons, we obtained simultaneous recordings of local field potentials from the CA1 stratum radiatum and demonstrated that the 3-Hz oscillations of CA2 neurons were not correlated with CA1 field potentials. In tetrodotoxin-perfused acute hippocampal slices, the membrane potentials of CA2 pyramidal cells were not preferentially entrained to 3-Hz sinusoidal current inputs, which suggest that intracellular 3-Hz oscillations reflect the neuronal dynamics of the surrounding networks. © 2016 Wiley Periodicals, Inc.

[Antithrombin resistance: a new mechanism of inherited thrombophilia].

Venous thromboembolism is a multifactorial disease resulting from complex interactions among genetic and environmental factors. To date, numerous genetic defects have been found in families with hereditary thrombophilia, but there may still be many undiscovered causative gene mutations. We investigated a possible causative gene defect in a large Japanese family with inherited thrombophilia, and found a novel missense mutation in the prothrombin gene (p.Arg596Leu) resulting in a variant prothrombin (prothrombin Yukuhashi). The mutant prothrombin had moderately lower activity than wild type prothrombin in clotting assays, but formation of the thrombin-antithrombin (TAT) complex was substantially impaired resulting in prolonged thrombin activity. A thrombin generation assay revealed that the peak activity of the mutant prothrombin was fairly low, but its inactivation was extremely slow in reconstituted plasma. The Leu596 substitution caused a gain-of-function mutation in the prothrombin gene, resulting in resistance to antithrombin and susceptibility to thrombosis. We also showed the effects of the prothrombin Yukuhashi mutation on the thrombomodulin-protein C anticoagulation system, recent development of a laboratory test detecting antithrombin resistance in plasma, and another antithrombin resistant mutation found in other thrombophilia families.

Ribavirin-induced intracellular GTP depletion activates transcription elongation in coagulation factor VII gene expression.

Coagulation FVII (Factor VII) is a vitamin K-dependent glycoprotein synthesized in hepatocytes. It was reported previously that FVII gene (F7) expression was up-regulated by ribavirin treatment in hepatitis C virus-infected haemophilia patients; however, its precise mechanism is still unknown. In the present study, we investigated the molecular mechanism of ribavirin-induced up-regulation of F7 expression in HepG2 (human hepatoma cell line). We found that intracellular GTP depletion by ribavirin as well as other IMPDH (inosine-5'-monophosphate dehydrogenase) inhibitors, such as mycophenolic acid and 6-mercaptopurine, up-regulated F7 expression. FVII mRNA transcription was mainly enhanced by accelerated transcription elongation, which was mediated by the P-TEFb (positive-transcription elongation factor b) complex, rather than by promoter activation. Ribavirin unregulated ELL (eleven-nineteen lysine-rich leukaemia) 3 mRNA expression before F7 up-regulation. We observed that ribavirin enhanced ELL3 recruitment to F7, whereas knockdown of ELL3 diminished ribavirin-induced FVII mRNA up-regulation. Ribavirin also enhanced recruitment of CDK9 (cyclin-dependent kinase 9) and AFF4 to F7. These data suggest that ribavirin-induced intracellular GTP depletion recruits a super elongation complex containing P-TEFb, AFF4 and ELL3, to F7, and modulates FVII mRNA transcription elongation. Collectively, we have elucidated a basal mechanism for ribavirin-induced FVII mRNA up-regulation by acceleration of transcription elongation, which may be crucial in understanding its pleiotropic functions in vivo.

[Discovery and prospects of a novel thrombophilia: antithrombin resistance].

Pathogenesis of venous thromboembolism (VTE) known to be complex and multifactorial process involves the interaction of acquired factors and genetic predisposing conditions. Deficiency of natural anticoagulant factors such as antithrombin (AT), protein C and protein S increases the risk of a VTE. Recently, we have reported novel mechanism of hereditary thrombosis in a Japanese family, in which AT resistance was associated with a missense mutation (p.Arg596Leu) in the prothrombin gene named prothrombin Yukuhashi. The mutant thrombin showed a low clotting activity, but a severely impaired inactivation by AT, resulting in a susceptibility to thrombosis. We have developed a new laboratory test to evaluate AT resistance in plasma. Prothrombin mutation causing AT resistance has found in Caucasian, not only in Japanese.

Aortic Replacement for a Crawford II Thoracoabdominal Aneurysm With a Right-Sided Descending Aorta: A Case Report.

A 53-year-old woman was diagnosed with a Crawford II thoracoabdominal aortic aneurysm involving the right-sided descending aorta. The patient underwent aortic replacement via a thoracoabdominal approach. The right-sided descending thoracic aortic aneurysm was excluded. The patient had a favorable postoperative course. The excluded thoracic aneurysm had completely thrombosed without intercostal inflow. The right-sided descending aorta is a rare malformation. The exclusion technique was appropriate because there was no retrograde flow from the intercostal arteries and the Adamkiewicz artery originated from the lumbar region.

Incomplete homonymous hemianopsia progressing to junctional scotoma due to a large internal carotid artery aneurysm: A case report.

Visual field disorders caused by cerebral aneurysms are diverse, nonspecific, and vary in their degree of compression. They should be distinguished from those caused by other common diseases, such as glaucoma.

Genotypes of Candida albicans isolated from healthy individuals and their distribution in patients with oral candidiasis.

For the study of Candida albicans genotypes involved in development of candidiasis, Candida albicans isolates were collected from healthy volunteers and patients with oral candidiasis and genotyped on the basis of 25S rDNA and microsatellite polymorphisms. In the microsatellite analysis using two microsatellite markers (CDC3 and CAI), 63 healthy volunteer isolates were classified into 35 genotypes (allelic relations to CDC3 alleles 1:2/CAI alleles 1:2), among which genotypes II (115:119/23:23), III (115:123/18:27), and V (123:127/32:41) were found at frequencies of 12.7%, 7.9%, and 7.9%, respectively. In 68 oral candidiasis isolates classified into 39 genotypes, genotypes II and III were identified in 4.4% and 20.6% of the isolates, respectively. The frequency of genotype III was higher in the candidiasis isolates than in the healthy isolates (p < 0.05). These results suggest that genotype III C. albicans assigned by CDC3/CAI is related to the development of oral candidiasis.

Comparison of the efficacy and invasiveness of manual and automated gonioscopy.

PURPOSE: To compare the efficacy and invasiveness of manual gonioscopy and automated 360-degree gonioscopy. METHOD: Manual and automated gonioscopy were performed on 70 patients with glaucoma. Manual gonioscopy was performed by a glaucoma specialist and an ophthalmology resident, and automated gonioscopy (GS-1) was performed by orthoptists. We compared the examination time for acquiring gonioscopic images (GS-1: 16 directions; manual gonioscopy: 8 directions). Furthermore, we compared the pain and discomfort scores during the examination using the Individualized Numeric Rating Scale. Among the images acquired by automated gonioscopy, we also evaluated the percentages of acquired images that could be used to determine the angle opening condition. RESULTS: The examination time was not significantly different between manual (80.2±28.7) and automated gonioscopy (94.7±82.8) (p = 0.105). The pain score of automated gonioscopy (0.22±0.59) was significantly lower than that of manual gonioscopy (0.55±1.11) (p = 0.025). The discomfort score was not significantly different between manual (1.34±1.90) and automated gonioscopy (1.06±1.50) (p = 0.165). Automated gonioscopy successfully acquired clear gonioscopic images in 93.4% of the total images. CONCLUSION: Automated gonioscopy is comparable in examination time and invasiveness to manual gonioscopy and may be useful for 360-degree iridocorneal angle evaluation.

A case of functional visual loss diagnosed through bilateral randomized visual field testing with a trick method.

Purpose: To report a case of functional visual loss (FVL) diagnosed through bilateral randomized visual field testing using Imo vifa with a trick method. Observations: A 27-year-old man complained of visual field abnormality in his left eye after falling from a height of 4 m. The left eye had a best-corrected visual acuity (BCVA) of 20/16 and a critical flicker frequency (CFF) of 44.5 Hz at the first visit. Commotio retinae was observed in the inferior retina of the left eye, and the pupillary light reflex was normal. Computed tomography and magnetic resonance imaging of the head revealed no abnormalities. However, the Goldmann perimeter (GP) showed constriction of visual field in the left eye. Since traumatic optic neuropathy was suspected initially; therefore, two courses of methylprednisolone pulse therapy were administered. However, the BCVA and CFF gradually worsened to 20/200 and 14 Hz, respectively. Nevertheless, his pupillary light reflex was still normal, and GP showed a spiral visual field. Thus, we suspected that this was a case of FVL and performed bilateral randomized visual field testing using Imo vifa in three steps as a trick method. In the first step, we performed the normal method for bilateral randomized visual field testing. In the second and third steps, we explained to the patient that only the right or left eye would be examined on purpose; bilateral randomized visual field testing was then performed. The results of examinations revealed left homonymous hemianopsias and normal and concentric contraction of the visual field in both eyes. These results could not be explained by organic disease, and the patient was diagnosed with FVL. Conclusions and Importance: Bilateral randomized visual field testing using Imo vifa with a trick method was useful for diagnosing FVL.

Primary leiomyosarcoma of the inferior vena cava in a pediatric case: a case report and literature review.

BACKGROUND: Leiomyosarcoma is classified as a soft tissue sarcoma. In adults, leiomyosarcoma is the most common malignancy affecting the vascular system; however, vascular leiomyosarcoma in children is extremely rare as most pediatric soft tissue tumors are rhabdomyosarcomas. The survival rate is very low, and incomplete resection is a poor prognostic factor. There is also a high rate of distant recurrence, with the lungs and liver being the most common sites of metastasis. There is no established effective chemotherapy, and complete surgical resection is the only potentially curative treatment for leiomyosarcoma. CASE PRESENTATION: A 15-year-old female patient with no significant medical history presented with severe upper abdominal pain and was admitted. Abdominal contrast-enhanced computed tomography and magnetic resonance imaging showed a large retroperitoneal tumor protruding into the lumen of the inferior vena cava behind the liver and multiple small nodules, and metastasis to the liver was suspected. The tumor was 6 × 4 × 5 cm in diameter, located just behind the hepatic hilar structures, and was suspected to infiltrate into the right portal vein. The tumor was diagnosed as a leiomyosarcoma through an open tumor biopsy. As the multiple liver metastases were located only in the right lobe of the liver on imaging, we performed tumor resection with right hepatectomy and replacement of the inferior vena cava (IVC). The postoperative course was uneventful; however, on postoperative day 51, distant metastatic recurrences were found in the remaining liver and right lung. The patient was immediately started on chemotherapy and trabectedin proved to be the most effective drug in the treatment regimen; however, severe side effects, such as hepatotoxicity, prevented timely administration, and the patient passed away 19 months after surgery. CONCLUSIONS: IVC resection and reconstruction combined with right hepatectomy were able to be safely performed even in a pediatric case. To improve the prognosis of leiomyosarcoma with multiple metastases, an effective treatment strategy combining surgical treatment and chemotherapy, including molecularly targeted drugs, should be established as early as possible.

Fatigue Potentially Reduces the Effect of Transcranial Magnetic Stimulation on Depression Following COVID-19 and Its Vaccination.

COVID-19's long-term effects, known as Long-COVID, present psychiatric and physical challenges in recovered patients. Similarly, rare long-term post-vaccination side effects, resembling Long-COVID, are emerging (called Post-Vaccine). However, effective treatments for both conditions are scarce. Our clinical experience suggests that transcranial magnetic stimulation (TMS) often aids recovery in Long-COVID and Post-Vaccine patients. However, its effectiveness is reduced in patients with severe fatigue. Therefore, we retrospectively analysed Tokyo TMS Clinic's outpatient records (60 in total; mean age, 38 years) to compare Long-COVID and post-vaccine patients' characteristics and symptoms, assess the impact of TMS on their symptoms, and investigate the role of fatigue in depression recovery with TMS. The primary outcome was the regression coefficient of the initial fatigue score on depression score improvement using TMS. Secondary outcomes included psychiatric/physical scores before and after TMS and their improvement rates. We found no differences in the initial symptoms and background factors between Long-COVID and Post-Vaccine patients. After ten TMS sessions, all psychiatric and physical symptom scores improved significantly. TMS improves depression, insomnia, anxiety, and related neuropsychiatric symptoms, which were the primary complaints in this study. Thus, we conclude that TMS improves depression and anxiety. The effectiveness of TMS in treating depression in Long-COVID and Post-Vaccine patients decreased as fatigue severity increased. In conclusion, TMS relieved depressive symptoms following COVID-19 and vaccination; however, fatigue may hinder its effectiveness.

Protocol for analysis of integrin-mediated cell adhesion of lateral plate mesoderm cells isolated from zebrafish embryos.

Lateral plate mesoderm (LPM) cells differentiate into various cell types including endothelial and hematopoietic cells. In zebrafish embryos, LPM cells migrate toward the midline along the ventral surfaces of somites during which their cell fate specification depends upon efficient integrin-mediated cell adhesion and migration. Herein, we present a protocol for analysis of integrin-mediated cell adhesion of LPM cells isolated from zebrafish embryos. This allows the study of the molecular mechanisms underlying integrin activation required for LPM cell fate specification. For complete details on the use and execution of this protocol, please refer to Rho et al. (2019).

A case of sigmoidectomy for sigmoid colon cancer with severe pulmonary arterial hypertension associated with mixed tissue connected disease: A case report.

INTRODUCTION: Patients with mixed connective tissue disease (MCTD) have higher rates of pulmonary arterial hypertension (PAH) than the general population. PAH is a risk for perioperative respiratory and heart failure, and marked edema of colonic stoma after sigmoidectomy. We report a case of sigmoidectomy for sigmoid colon cancer in a patient with PAH associated with MCTD for whom perioperative treatment was planned to control pulmonary arterial pressure (PAP), and a surgical strategy to avoid complications attributable to PAH and MCTD was employed. CASE PRESENTATION: A 52-year-old woman with sigmoid cancer and severe PAH associated with MCTD underwent surgery. We controlled PAH by using intravenous epoprostenol. We selected open surgery without laparoscopy and Hartmann's operation. After surgery, severe perioperative complications were not detected, and the patient discharged from hospital 17 days after the operation. DISCUSSION: During surgery under general anesthesia, the mortality rate of PAH is high because of heart and respiratory failure. We planned to switch the PAH treatment from an oral agent to intravenous epoprostenol only in the preoperative period, and selected open surgery. We ligated the inferior mesenteric artery (IMA) and inferior mesenteric vein (IMV) below the branch of LCA to avoid marked edema of stoma. Consequently, we could avoid severe intraoperative and postoperative complications. CONCLUSIONS: Controlling PAP using epoprostenol, open surgery, stoma and the ligation level for the IMA and IMV preventing are important to avoid perioperative complications of sigmoid colon cancer complicated by severe PAH.

Association of Global Coagulation Profiles With Cardiovascular Risk Factors and Atherosclerosis: A Sex Disaggregated Analysis From the BioHEART-CT Study.

Background Although the association between dysregulated coagulation and atherosclerosis is well recognized, individual assays have been of minimal value in understanding disease susceptibility. Here we investigated the association of global coagulation profiles with coronary artery disease with consideration of sex differences. Methods and Results The study included patients from the BioHEART-CT (The BioHEART Study: Assessing Patients With Suspected Cardiovascular Disease for New Disease Markers and Risk Factors) biobank who had computed tomography coronary angiograms scored for coronary artery calcium score (CACS) and Gensini score. The cohort included 206 adult patients who were referred for clinically indicated computed tomography coronary angiography and had a median of 2 major cardiac risk factors; 50% were women and the average age was 62.6 years (±9.9 years). The overall hemostatic potential (OHP) and calibrated automated thrombography generation assays were performed on platelet-poor plasma. CACS and Gensini score in men were significantly correlated in bivariate analysis with measures from the OHP assay, and regression models predicting disease severity by CACS or Gensini score were improved by adding the OHP assay variables in men but not in women. The calibrated automated thrombography generation assay demonstrated a more hypercoagulable profile in women than in men. The OHP assay showed hypercoagulable profiles in women with hyperlipidemia and men with obesity. Conclusions The OHP assay identified hypercoagulable profiles associated with different risk factors for each sex and was associated with CACS and Gensini score severity in men, emphasizing the associations between increased fibrin generation and reduced fibrinolysis with cardiac risk factors and early atherosclerosis. Registration Information www.anzctr.org.au. Identifier: ACTRN12618001322224.

Prediction of anterior chamber volume after implantation of posterior chamber phakic intraocular lens.

PURPOSE: To predict the anterior chamber volume (ACV) after implantable collamer lens (ICL) implantation based on ICL size and parameters of anterior segment optical coherence tomography (AS-OCT). DESIGN: Retrospective study. METHODS: This study included 222 eyes of 222 patients who underwent ICL implantation at Nagoya Eye Clinic. The patients were divided into two groups: prediction group, for creating the prediction equation (148 eyes, mean age: 32.11 ± 8.04 years), and verification group, for verifying the equation (74 eyes, mean age: 33.03 ± 6.74 years). The angle opening distance (AOD), anterior chamber width (ACW), ACV, anterior chamber depth, lens vault, angle-to-angle distance, angle recess area, and trabecular iris space area were calculated using AS-OCT. A stepwise multiple regression analysis was performed. After the creation of the prediction equation, its accuracy was verified in the verification group. RESULTS: The ACV, AOD750, ACW, and ICL size were selected as explanatory variables to predict postoperative ACV. Mean predicted (114.2 ± 21.83 mm3) and actual postoperative ACVs (116.1 ± 25.41 mm3) were not significantly different (P = 0.269); absolute error was 10.59 ± 9.13 mm3. In addition, there was high correlation between actual and predictive ACV (adjusted R2 = 0.6996, p < 0.0001). Bland-Altman plot revealed that there was no addition or proportional error between predicted and actual postoperative ACV. CONCLUSION: Postoperative ACV was accurately predicted using AS-OCT parameters and ICL size. This prediction equation may be useful for making decisions regarding ICL size.

Keratoconus Screening Using Values Derived From Auto-Keratometer Measurements: A Multicenter Study.

PURPOSE: Screening of early-stage keratoconus using auto-keratometer parameters. DESIGN: Evaluation of a screening approach. METHODS: At 5 major centers in Japan, we enrolled 123 eyes of 123 patients with Amsler-Krumeich classification stage 1 (<50 years of age [average 26.36 ± 8.68 years]; 84/39 male/female) and 205 eyes of 205 healthy subjects (average age 26.20 ± 7.34 years, 139/66 male/female). Participants were divided 2:1 into a prediction group and an application group. In the prediction group, multivariate logistic regression analysis was performed with keratoconus diagnosis as the dependent variable, and auto-keratometer parameters including average K, steep K, flat K, astigmatism, and astigmatic axis (no, with-the-rule, against-the-rule, and oblique) as independent variables. The diagnostic probability determined by regression analysis was defined as the keratometer keratoconus index. The cutoff value was determined from the receiver operating characteristic curve. This prediction equation was evaluated in the application group. Our primary outcome measure was the accuracy of the prediction equation for discriminating keratoconus from normal eyes. RESULTS: The selected explanatory variables were steep K (partial regression coefficient [ß] 1.284, odds ratio [OR] 3.610), flat K (ß -0.618, OR 0.539), and with-the-rule astigmatism (ß -3.163, OR 0.042). The area under the receiver operating characteristic curve of keratometer keratoconus index was 0.90, which was significantly better than individual parameters (P < .001). The sensitivity and specificity values in the application group were 85.0% and 86.7%, respectively. CONCLUSIONS: Although the sensitivity/specificity was not high, the new prediction equation using auto-keratometer-derived parameters enabled better discrimination of early-stage keratoconus than the isolated parameters.

Osteonecrosis associated with short-term oral administration of bisphosphonate.

Orally administered bisphosphonates are generally considered a first-line medication for treatment of osteoporosis. As a side effect of bisphosphonates, osteonecrosis of the jaws (ONJ) has been reported worldwide. Most reports of osteonecrosis are attributed to the use of bisphosphonates administered intravenously for the treatment of bone disorders such as multiple myeloma. However, osteonecrosis has also been diagnosed in patients receiving oral bisphosphonates. The management of a patient with spontaneous removable partial denture prosthesis-related ONJ associated with short-term oral administration of bisphosphonate is described.

Recurrent prolapse of toric implantable collamer lens after blunt ocular trauma under mesopic conditions.

Implantable collamer lenses (ICL) carry a risk of prolapse caused by direct ocular trauma, especially in mesopic conditions and when oversized ICLs are implanted. We recommend early surgical repositioning, as well as patient education that encourages goggle use during active sports.