Novel role of LLGL2 silencing in autophagy: reversing epithelial-mesenchymal transition in prostate cancer.

PURPOSE: Prostate cancer (PCa) is a major urological disease that is associated with significant morbidity and mortality in men. LLGL2 is the mammalian homolog of Lgl. It acts as a tumor suppressor in breast and hepatic cancer. However, the role of LLGL2 and the underlying mechanisms in PCa have not yet been elucidated. Here, we investigate the role of LLGL2 in the regulation of epithelial-mesenchymal transition (EMT) in PCa through autophagy in vitro and in vivo. METHODS: PC3 cells were transfected with siLLGL2 or plasmid LLGL2 and autophagy was examined. Invasion, migration, and wound healing were assessed in PC3 cells under autophagy regulation. Tumor growth was evaluated using a shLLGL2 xenograft mouse model. RESULTS: In patients with PCa, LLGL2 levels were higher with defective autophagy and increased EMT. Our results showed that the knockdown of LLGL2 induced autophagy flux by upregulating Vps34 and ATG14L. LLGL2 knockdown inhibits EMT by upregulating E-cadherin and downregulating fibronectin and α-SMA. The pharmacological activation of autophagy by rapamycin suppressed EMT, and these effects were reversed by 3-methyladenine treatment. Interestingly, in a shLLGL2 xenograft mouse model, tumor size and EMT were decreased, which were improved by autophagy induction and worsened by autophagy inhibition. CONCLUSION: Defective expression of LLGL2 leads to attenuation of EMT due to the upregulation of autophagy flux in PCa. Our results suggest that LLGL2 is a novel target for alleviating PCa via the regulation of autophagy.

Corrigendum: Elevated IFNA1 and suppressed IL12p40 associated with persistent hyperinflammation in COVID-19 pneumonia.

[This corrects the article DOI: 10.3389/fimmu.2023.1101808.].

Elevated IFNA1 and suppressed IL12p40 associated with persistent hyperinflammation in COVID-19 pneumonia.

Introduction: Despite of massive endeavors to characterize inflammation in COVID-19 patients, the core network of inflammatory mediators responsible for severe pneumonia stillremain remains elusive. Methods: Here, we performed quantitative and kinetic analysis of 191 inflammatory factors in 955 plasma samples from 80 normal controls (sample n = 80) and 347 confirmed COVID-19 pneumonia patients (sample n = 875), including 8 deceased patients. Results: Differential expression analysis showed that 76% of plasmaproteins (145 factors) were upregulated in severe COVID-19 patients comparedwith moderate patients, confirming overt inflammatory responses in severe COVID-19 pneumonia patients. Global correlation analysis of the plasma factorsrevealed two core inflammatory modules, core I and II, comprising mainly myeloid cell and lymphoid cell compartments, respectively, with enhanced impact in a severity-dependent manner. We observed elevated IFNA1 and suppressed IL12p40, presenting a robust inverse correlation in severe patients, which was strongly associated with persistent hyperinflammation in 8.3% of moderate pneumonia patients and 59.4% of severe patients. Discussion: Aberrant persistence of pulmonary and systemic inflammation might be associated with long COVID-19 sequelae. Our comprehensive analysis of inflammatory mediators in plasmarevealed the complexity of pneumonic inflammation in COVID-19 patients anddefined critical modules responsible for severe pneumonic progression.

Antitumor Potential of Sericite Treatment Mediated by Cell Cycle Arrest in Triple-Negative MDA-MB231 Breast Cancer Cells.

Breast cancer is the most common cancer and the leading cause of cancer-related mortality among females worldwide. Triple-negative breast cancer (TNBC) accounts for about 10-15% of all breast cancers and is usually more aggressive and has a poorer prognosis. Sericite has been known to have antitumor and immune-stimulatory effects. Although the chemopreventive potential of sericite has been demonstrated in other cancers, its molecular pathways in TNBC still require investigation. Thus, in the present study, the antitumor mechanism of sericite against MDA-MB231 breast cancer cells was examined in vitro and in an in vivo xenograft mouse model. Sericite treatment reduced cell proliferation and cell proliferation marker proliferating cell nuclear antigen (PCNA) in MDA-MB231 cells. It also decreased the total cell number and arrested cells in the G0/G1 phase of the cell cycle with an increase in the phosphorylation of P53 and upregulation of cell cycle regulatory proteins P21 and P16. In addition, sericite treatment also induced apoptosis signaling, which was evident by the upregulation of apoptotic protein markers cleaved caspases 3 and 9. A reduction in reactive oxygen species (ROS), NADPH oxidase 4 (NOX4), p22phox, and heat shock proteins (HSPs) was also observed. Similar results were obtained in vivo with significantly reduced tumor volume in sericite-administered mice. Collectively, these findings suggest that sericite has antitumor potential based on its property to induce cell cycle arrest and apoptotic cell death and therefore could serve as a potential therapeutic agent and crucial candidate in anticancer drug development for TNBC.

Ref-1 protects against FeCl3-induced thrombosis and tissue factor expression via the GSK3ß-NF-κB pathway.

Arterial thrombosis and its associated diseases are considered to constitute a major healthcare problem. Arterial thrombosis, defined as blood clot formation in an artery that interrupts blood circulation, is associated with many cardiovascular diseases. Oxidative stress is one of many important factors that aggravates the pathophysiological process of arterial thrombosis. Apurinic/apyrimidinic endonuclease 1/redox factor-1 (Ref-1) has a multifunctional role in cells that includes the regulation of oxidative stress and anti-inflammatory function. The aim of this study was to investigate the therapeutic effect of adenovirus-mediated Ref-1 overexpression on arterial thrombosis induced by 60% FeCl3 solution in rats. Blood flow was measured to detect the time to occlusion, thrombus formation was detected by hematoxylin and eosin staining, reactive oxygen species (ROS) levels were detected by high-performance liquid chromatography, and the expression of tissue factor and other proteins was detected by Western blot. FeCl3 aggravated thrombus formation in carotid arteries and reduced the time to artery occlusion. Ref-1 significantly delayed arterial obstruction via the inhibition of thrombus formation, especially by downregulating tissue factor expression through the Akt-GSK3ß-NF-κB signaling pathway. Ref1 also reduced the expression of vascular inflammation markers ICAM-1 and VCAM1, and reduced the level of ROS that contributed to thrombus formation. The results showed that adenovirus-mediated Ref-1 overexpression reduced thrombus formation in the rat carotid artery. In summary, Ref-1 overexpression had anti-thrombotic effects in a carotid artery thrombosis model and could be a target for the treatment of arterial thrombosis.

Type A aortic dissection after 'zone 0' thoracic endovascular aortic repair for type 1 hybrid aortic arch replacement of arch aneurysm.

The recent rise in minimally invasive cardiovascular procedures is being accompanied by an increase in related complications. We report on an acute type A aortic dissection performed in an 82-year-old man 1 week after staged 'zone 0' hybrid thoracic endovascular aortic repair (TEVAR). Previously, the patient had undergone type I hybrid arch debranching and staged 'zone 0' TEVAR for an aortic arch aneurysm. 'Zone 0' TEVAR after type I hybrid debranching might increase the risk for aortic injury on the residual native aorta and should, therefore, be closely followed up to enable the early diagnosis of complications.

Surgical treatment for mediastinitis after endobronchial ultrasound-guided transbronchial needle aspiration: 2 case reports.

We report two cases of severe mediastinitis accompanied by abscess due to endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA), that were successfully treated by effective surgical drainage. A 68-year-old woman was referred to our hospital due to chest discomfort and high fever after EBUS-TBNA, and a 54-year-old man was referred due to general weakness, chills, and high fever after the same procedure. Both were diagnosed with EBUS-related mediastinitis and discharged after surgical treatment. Similar to previous reports, the importance of surgical procedures for mediastinitis caused by EBUS-TBNA was suggested. Further research and establishment of guidelines on this matter is necessary.

Thoracic Outlet Syndrome Induced by Huge Lipoma: A Case Report.

Thoracic outlet syndrome is a relatively well known disease. Other than trauma, this disease is mostly caused by anatomical structures that cause vascular or neural compression. The cause of thoracic outlet syndrome is diverse; however, there are only few reports of thoracic outlet syndrome caused by lipoma in the pectoralis minor space. We report a case of compression of the lower trunk of brachial plexus in which a large lipoma that developed in the pectoral minor space grew into the subclavicular space, along with a review of literature.

Novel Asymmetrical Linear Stapler (NALS) for pathologic evaluation of true resection margin tissue.

BACKGROUND: The use of limited resection for lung cancer has increased with the accumulation of knowledge about early lung cancer. To decrease locoregional recurrence after a limited resection, it is important to confirm R0 resection at the true resection margin. In this study, we report a novel linear stapler that preserves the true resection margin tissue after organ resection. METHODS: We used a Novel Asymmetrical Linear Stapler (NALS) made by Meditulip. On the resected organ side of NALS, there is a single row of titanium fasteners. To verify the utility of NALS and to compare its preservation of the resection margin tissue to a conventional stapler, we performed wedge resection of the lung in a porcine animal model and examined the pathology of the true resection margin. RESULTS: Using NALS, we successfully divided and closed the lung tissues, as with the conventional stapler. There was no bleeding on either side or no air leakage from the remnant stapled tissue. The distance between the cutting edge and the titanium fasteners was 3.10 mm with NALS, which was sufficient to resect the true resection margin tissue for pathology evaluation. There was no squeezing artifact at the true resection margin on microscopic evaluation with NALS. With the conventional stapler, it is difficult to evaluate the pathology at the true resection margin due to the severe squeezing artifact. CONCLUSIONS: NALS preserves the true resection margin tissue and thus should be useful for evaluating the resection margin with a frozen section biopsy in oncology surgery.

Electromagnetic navigation bronchoscopy-Chungnam National University Hospital experience.

BACKGROUND: To find small pulmonary nodules or ground grass nodules (GGNs) with video-assisted thoracoscopic surgery (VATS) is very difficult. There are several conventional methods to localize small nodules or GGNs, which require additional radiation exposure and may cause some complications such as pneumothorax or hemothorax. We aimed to evaluate the effectiveness and feasibility of electromagnetic navigational bronchoscopy-guided pulmonary localization in a minimally invasive thoracic surgery field. METHODS: We retrospectively reviewed the medical records from a prospectively collected database of the patients who underwent ENB procedure for biopsy and/or localization of pulmonary resection at the Chungnam National University Hospital from January 2017 to January 2018. RESULTS: A total of 37 ENB-guided dye-markings or biopsies for 37 lesions in 30 patients were performed. Thirty-two ENB-guided localizations using dye-marking for resection were performed in 25 patients. The median nodule size was 9 mm (IQR: 7-13 mm), and the median distance from the pleura was 6 mm (IQR: 3-10 mm). The failure of an ENB-guided localization was noted in 4 cases (12.5%). There was no major complication noted with the procedure, and just two patients showed mild intrabronchial bleeding stopped spontaneously. The most common lobar location was right lower lobe (11 cases, 34.4%), and all cases of localization failure were right lower lobe. A pathologic diagnosis was obtained from surgically resected specimen (not from ENB biopsy: 32 of 32 localizations, 100%), neoplastic lesions were 23 cases (72%). Of them, a primary lung cancer and metastatic lung cancer were noted in 11 cases, and in 11cases, respectively. All margins of the nodules were negative. CONCLUSIONS: The ENB-guided dye localization by a well-trained thoracic surgeon enables accurate intraoperative identification of GGN or a small pulmonary nodule, with minimal complications and enables minimally invasive surgery including single port surgery.

Prognostic significance of nuclear factor of activated T-cells 5 expression in non-small cell lung cancer patients who underwent surgical resection.

BACKGROUND: Nuclear factor of activated T-cells 5 (NFAT5) is known to be correlated with migration or invasion of tumor cells based on previous in vitro studies. The aim of this study was to analyze the relationship between NFAT5 expression and clinical prognosis in non-small cell lung cancer (NSCLC) patients who underwent surgical resection. MATERIALS AND METHODS: A total of 92 NSCLC patients who underwent surgical resection were enrolled. The tissue microarray core was obtained from surgically resected tumor specimens. NFAT5 expression was evaluated by immunohistochemistry. Relationships of NFAT5 expression with disease recurrence, overall survival, and disease-free survival (DFS) were analyzed. RESULTS: The mean age of 92 patients was 63.7 y. The median follow-up duration was 63.3 mo. Fifty-one (55%) patients exhibited positive expression of NFAT5. Disease recurrence in the NFAT5-positive group was significantly (P = 0.022) higher than that in the NFAT5-negative group. NFAT5-positive expression (odds ratio: 2.632, 95% confidence interval: 1.071-6.465, P = 0.035) and pathologic N stage (N1-2 versus N0; odds ratio: 3.174, 95% confidence interval: 1.241-8.123, P = 0.016) were independent and significant risk factors for disease recurrence. DFS of the NFAT5-positive group was significantly worse than that of the NFAT5-negative group (89.7 versus 48.7 mo, P = 0.011). A multivariate analysis identified NFAT5 expression (P < 0.029) as a significant independent risk factor for DFS of patients with postoperative pathologic T and N stages (P < 0.001 and P = 0.017, respectively). CONCLUSIONS: NFAT5 expression is a useful prognostic biomarker for NSCLC patients who underwent surgical resection.

A Life-Threatening Bronchogenic Cyst.

A bronchogenic cyst causing cardiac tamponade is a rare condition. We report an unusual case of a bronchogenic cyst that caused cardiac tamponade. A 49-year-old female patient presented at our emergency room with complaints of palpitations and shortness of breath that had lasted for 5 days preceding the visit. Echocardiography revealed a very large cystic mass compressing the left a trium posteriorly, and a large amount of pericardial effusion caused the diastolic collapse of the ventricles. Atrial fibrillation and aggravated dyspnea were observed, and the patient's vital signs were unstable after admission. We therefore performed an emergency operation. The bronchogenic cyst was resected by thoracotomy and the patient was discharged 12 days after the operation without any complications over 5 years of follow-up.

CR6-Interacting Factor 1 Deficiency Impairs Vascular Function by Inhibiting the Sirt1-Endothelial Nitric Oxide Synthase Pathway.

AIMS: Mitochondrial dysfunction has emerged as a major contributing factor to endothelial dysfunction and vascular disease, but the key mechanisms underlying mitochondrial dysfunction-induced endothelial dysfunction remain to be elucidated. In this study, we aim at determining whether mitochondrial dysfunction in endothelial cells plays a key role in vascular disease, by examining the phenotype of endothelial-specific CR6-interacting factor 1 (CRIF1) knockout mice. We also used siRNA-mediated downregulation of CRIF1 gene in the endothelial cells to study about the in vitro pathophysiological underlying mechanisms. RESULTS: Downregulation of CRIF1 in endothelial cells caused disturbances of mitochondrial oxidative phosphorylation complexes and membrane potential, leading to enhanced mitochondrial reactive oxygen species production. Gene silencing of CRIF1 results in decreased SIRT1 expression along with increased endothelial nitric oxide synthase (eNOS) acetylation, leading to reduced nitric oxide production both in vitro and in vivo. Endothelium-dependent vasorelaxation of aortic rings from CRIF1 knockout (KO) mice was considerably less than in wild-type mice, and it was partially recovered by Sirt1 overexpression in CRIF1 KO mice. INNOVATION: Our results show for the first time a relationship between mitochondrial dysfunction and impaired vascular function induced in CRIF1 deficiency conditions and also the possible underlying pathway involved. CONCLUSION: These findings indicate that CRIF1 plays an important role in maintaining mitochondrial and endothelial function through its effects on the SIRT1-eNOS pathway. Antioxid. Redox Signal. 27, 234-249.

Extracorporeal Membrane Oxygenation in a 1,360-g Premature Neonate after Repairing Total Anomalous Pulmonary Venous Return.

With advancements in complex repairs in neonates with complicated congenital heart diseases, extracorporeal membrane oxygenation (ECMO) has been increasingly used as cardiac support. ECMO has also been increasingly used for low birth weight (LBW) or very low birth weight (VLBW) neonates. However, since prematurity and LBW are risk factors for ECMO, the appropriate indications for neonates with LBW, especially VLBW, are under dispute. We report a case of ECMO performed in a 1,360-g premature infant with VLBW due to cardiopulmonary bypass weaning failure after repairing infracardiac total anomalous pulmonary venous return.

Classic Peripheral Signs of Subacute Bacterial Endocarditis.

A 50-year-old female patient with visual disturbances was referred for further evaluation of a heart murmur. Fundoscopy revealed a Roth spot in both eyes. A physical examination showed peripheral signs of infective endocarditis, including Osler nodes, Janeway lesions, and splinter hemorrhages. Our preoperative diagnosis was subacute bacterial endocarditis with severe aortic regurgitation. The patient underwent aortic valve replacement and was treated with intravenous antibiotics for 6 weeks postoperatively. The patient made a remarkable recovery and was discharged without complications. We report this case of subacute endocarditis with all 4 classic peripheral signs in a patient who presented with visual disturbance.

In Vivo Neuroprotective Effect of Histidine-Tryptophan-Ketoglutarate Solution in an Ischemia/Reperfusion Spinal Cord Injury Animal Model.

BACKGROUND: Paraplegia is a devastating complication following operations on the thoracoabdominal aorta. We investigated whether histidine-tryptophan-ketoglutarate (HTK) solution could reduce the extent of ischemia/reperfusion (IR) spinal cord injuries in a rat model using a direct delivery method. METHODS: Twenty-four Sprague-Dawley male rats were randomly divided into four groups. The sham group (n=6) underwent a sham operation, the IR group (n=6) underwent only an aortic occlusion, the saline infusion group (saline group, n=6) underwent an aortic occlusion and direct infusion of cold saline into the occluded aortic segment, and the HTK infusion group (HTK group, n=6) underwent an aortic occlusion and direct infusion of cold HTK solution into the occluded aortic segment. An IR spinal cord injury was induced by transabdominal clamping of the aorta distally to the left renal artery and proximally to the aortic bifurcation for 60 minutes. A neurological evaluation of locomotor function was performed using the modified Tarlov score after 48 hours of reperfusion. The spinal cord was harvested for histopathological and immunohistochemical examinations. RESULTS: The spinal cord IR model using direct drug delivery in rats was highly reproducible. The Tarlov score was 4.0 in the sham group, 1.17±0.75 in the IR group, 1.33±1.03 in the saline group, and 2.67±0.81 in the HTK group (p=0.04). The histopathological analysis of the HTK group showed reduced neuronal cell death. CONCLUSION: Direct infusion of cold HTK solution into the occluded aortic segment may reduce the extent of spinal cord injuries in an IR model in rats.

Voice Change Due to Paratracheal Air Cysts.

Paratracheal air cysts are a rare entity in which cystic formation occurs adjacent to the trachea. Most patients with paratracheal air cysts are asymptomatic, and the cysts are detected incidentally on chest radiograph or computed tomography (CT) scan. Most symptomatic patients complain of pulmonary symptoms or repeated respiratory infection. Rarely, the air cysts can lead to paralysis of the recurrent laryngeal nerve as a result of direct compression. We report a case of a 59-year-old male patient who presented with voice change, and the cause was identified as paratracheal air cysts on a chest CT scan. Surgical resection via video-assisted mediastinoscopy was performed, and the voice recovered immediately after the operation.

Life-Threatening Congenital Cystic Adenomatoid Malformation in the Premature Neonate.

Congenital cystic adenomatoid malformation is a rare, but well-known disease. It can be managed conservatively in patients without symptoms or require surgical removal when symptomatic. The surgical option of choice is en bloc resection of the affected lesion. We report an experience of life-threatening congenital cystic adenoid malformation in a low-birth-weight (1,590 g) premature neonate who was successfully treated with a lobectomy of the lung.

Outcomes of the Tower Crane Technique with a 15-mm Trocar in Primary Spontaneous Pneumothorax.

BACKGROUND: Video-assisted thoracoscopic surgery (VATS) pulmonary wedge resection has emerged as the standard treatment for primary spontaneous pneumothorax. Recently, single-port VATS has been introduced and is now widely performed. This study aimed to evaluate the outcomes of the Tower crane technique as novel technique using a 15-mm trocar and anchoring suture in primary spontaneous pneumothorax. METHODS: Patients who underwent single-port VATS wedge resection in Chungnam National University Hospital from April 2012 to March 2014 were enrolled. The medical records of the enrolled patients were reviewed retrospectively. RESULTS: A total of 1,251 patients were diagnosed with pneumothorax during this period, 270 of whom underwent VATS wedge resection. Fifty-two of those operations were single-port VATS wedge resections for primary spontaneous pneumothorax performed by a single surgeon. The median age of the patients was 19.3±11.5 years old, and 43 of the patients were male. The median duration of chest tube drainage following the operation was 2.3±1.3 days, and mean postoperative hospital stay was 3.2±1.3 days. Prolonged air leakage for more than three days following the operation was observed in one patient. The mean duration of follow-up was 18.7±6.1 months, with a recurrence rate of 3.8%. CONCLUSION: The tower crane technique with a 15-mm trocar may be a promising treatment modality for patients presenting with primary spontaneous pneumothorax.

IDH2 deficiency impairs mitochondrial function in endothelial cells and endothelium-dependent vasomotor function.

Mitochondrial NADP(+)-dependent isocitrate dehydrogenase (IDH2) plays an essential role protecting cells against oxidative stress-induced damage. A deficiency in IDH2 leads to mitochondrial dysfunction and the production of reactive oxygen species (ROS) in cardiomyocytes and cancer cells. However, the function of IDH2 in vascular endothelial cells is mostly unknown. In this study the effects of IDH2 deficiency on mitochondrial and vascular function were investigated in endothelial cells. IDH2 knockdown decreased the expression of mitochondrial oxidative phosphorylation (OXPHOS) complexes I, II and III, which lead to increased mitochondrial superoxide. In addition, the levels of fission and fusion proteins (Mfn-1, OPA-1, and Drp-1) were significantly altered and MnSOD expression also was decreased by IDH2 knockdown. Furthermore, knockdown of IDH2 decreased eNOS phosphorylation and nitric oxide (NO) concentration in endothelial cells. Interestingly, treatment with Mito-TEMPO, a mitochondrial-specific superoxide scavenger, recovered mitochondrial fission-fusion imbalance and blunted mitochondrial superoxide production, and reduced the IDH2 knockdown-induced decrease in MnSOD expression, eNOS phosphorylation and NO production in endothelial cells. Endothelium-dependent vasorelaxation was impaired, and the concentration of bioavailable NO decreased in the aortic ring in IDH2 knockout mice. These findings suggest that IDH2 deficiency induces endothelial dysfunction through the induction of dynamic mitochondrial changes and impairment in vascular function.