Overexpression of Activin Receptor-Like Kinase 1 in Endothelial Cells Suppresses Development of Arteriovenous Malformations in Mouse Models of Hereditary Hemorrhagic Telangiectasia.

RATIONALE: Hereditary hemorrhagic telangiectasia (HHT) is a genetic disease caused by mutations in ENG, ALK1, or SMAD4. Since proteins from all 3 HHT genes are components of signal transduction of TGF-ß (transforming growth factor ß) family members, it has been hypothesized that HHT is a disease caused by defects in the ENG-ALK1-SMAD4 linear signaling. However, in vivo evidence supporting this hypothesis is scarce. OBJECTIVE: We tested this hypothesis and investigated the therapeutic effects and potential risks of induced-ALK1 or -ENG overexpression (OE) for HHT. METHODS AND RESULTS: We generated a novel mouse allele (ROSA26Alk1) in which HA (human influenza hemagglutinin)-tagged ALK1 and bicistronic eGFP expression are induced by Cre activity. We examined whether ALK1-OE using the ROSA26Alk1 allele could suppress the development of arteriovenous malformations (AVMs) in wounded adult skin and developing retinas of Alk1- and Eng-inducible knockout (iKO) mice. We also used a similar approach to investigate whether ENG-OE could rescue AVMs. Biochemical and immunofluorescence analyses confirmed the Cre-dependent OE of the ALK1-HA transgene. We could not detect any pathological signs in ALK1-OE mice up to 3 months after induction. ALK1-OE prevented the development of retinal AVMs and wound-induced skin AVMs in Eng-iKO as well as Alk1-iKO mice. ALK1-OE normalized expression of SMAD and NOTCH target genes in ENG-deficient endothelial cells (ECs) and restored the effect of BMP9 (bone morphogenetic protein 9) on suppression of phosphor-AKT levels in these endothelial cells. On the other hand, ENG-OE could not inhibit the AVM development in Alk1-iKO models. CONCLUSIONS: These data support the notion that ENG and ALK1 form a linear signaling pathway for the formation of a proper arteriovenous network during angiogenesis. We suggest that ALK1 OE or activation can be an effective therapeutic strategy for HHT. Further research is required to study whether this therapy could be translated into treatment for humans.

Esculetin has therapeutic potential via the proapoptotic signaling pathway in A253 human submandibular salivary gland tumor cells.

Esculetin is a natural lactone that is commonly derived from coumarins. According to previous experiments using human cancer cells, esculetin has potent antitumor activity; it also inhibits proliferation and induces the apoptosis of cancer cells. In the present study, the anti-proliferative effect of esculetin on the submandibular salivary gland tumor cell line, A253, was evaluated via in vitro and in vivo analyses. Furthermore, the anti-cancer effects of esculetin in A253 cells and a xenograft model of salivary gland tumors were determined using 3-(4,5-dimethylthiazol)-2,5-diphenyltetrazolium bromide and TUNEL assay, apoptosis protein array, quantitative polymerase chain reaction and western blot analysis. Esculetin (50-150 µM) was demonstrated to have an anti-proliferative effect in the A253 cell line in vitro; this observed effect was dependent on the dose and duration of treatment. Esculetin also increased the levels of Bax, cleaved caspase-3, cleaved-9 and cleaved poly (ADP-ribose) polymerase apoptosis-related proteins, and decreased the expression levels of the Bcl-2 anti-apoptotic protein. With respect to apoptosis regulation, esculetin significantly decreased the proliferation of tumor cells in a xenograft model (100 mg/kg/day) for 18 days. Overall, esculetin could be a potential oral anticancer drug against salivary gland cancer.

Effects of Rehabilitation in Bankart Lesion in Non-athletes: A Report of Three Cases.

A Bankart lesion is a common traumatic sports injury of the glenohumeral joint. When the shoulder is dislocated, the connective tissue surrounding the joint may tear or rupture. This study aimed to describe the initial dislocation, treatment, medical treatment, rehabilitation outcomes at 3 and 8 months, and return to daily life for three patients. Patient 1 was a 28-year-old male office worker who enjoyed fitness and weight training. His injury was sustained by a fall from his bicycle and subsequent performance of pull-up exercises. Surgery was recommended for repair of the anterior and inferior labrum tear. Reassessment of the surgery was performed after 2 months of rehabilitation. The patient was asked to exercise once a week at a rehabilitation center and to perform home-based exercises. In the first 2 months, the main exercise consisted of range of motion (ROM) exercises and stretching. Thereafter, muscle strengthening, using tubing, equipment and body weight exercises, and proprioceptive exercises were performed to gradually improve muscle strength and for return to daily activities. ROM progressively increased, with recovery of normal range at 2 months post-surgery. However, pain decreased only slightly, with a visual analog score of 6 out of 10 at 2 months post-surgery. At 8 months, the pain was bearable, without causing discomfort. Muscle strength remained almost unchanged initially but gradually recovered, although with a residual deficit of 20-30% in flexion, adduction and internal rotation. Considering the prolonged rehabilitation after surgery, future studies are warranted to analyze the long-term effects of non-surgical therapies by accumulating more cases and developing effective exercise rehabilitation programs.

What factors predict recurrence after an initial episode of primary spontaneous pneumothorax in children?

PURPOSE: Recurrence is the most common complication after an initial episode of primary spontaneous pneumothorax (PSP). However, preventive surgery in children remains a controversial issue. The purpose of this study was to determine predictive factors of recurrence to better inform early surgical referrals. METHODS: We retrospectively reviewed all consecutive patients under 18 years of age who conservatively treated for an initial episode of PSP between March 2005 and September 2011. RESULTS: One hundred fourteen patients were included in this study. The mean follow-up period was 43.1 months. Ipsilateral and contralateral recurrence developed in 47.3% and 14.0% of patients. The risk of ipsilateral recurrence for patients with or without air-containing lesions according to high-resolution computed tomography (HRCT) was 60.3% and 31.4%. In the multivariate analysis, the presence of air-containing lesions on HRCT scans and bullae on chest X-rays were independent risk factors for ipsilateral recurrence. CONCLUSION: The presence of bleb or bullae on HRCT scans or chest X-rays after an initial episode of PSP was significantly related to the ipsilateral recurrence in children. If the risk factors are clarified in further studies, hospital stays and the recurrence of PSP after the first episode could be reduced with early video-assisted thoracoscopic surgery.

Feasibility of optic nerve sheath diameter measured on initial brain computed tomography as an early neurologic outcome predictor after cardiac arrest.

OBJECTIVES: Few parameters are available to predict neurologic outcome of post-cardiac arrest patients in the early stage of treatment. Optic nerve sheath diameter (ONSD) has been used to indirectly assess intracranial pressure. This study evaluated whether ONSD, an additional parameter in initial brain computed tomography (CT) scans, can be an early predictor of neurologic outcome in post-cardiac arrest patients. METHODS: A total of 112 cardiac arrest patients between November 2012 and October 2013 were identified. Ninety-eight comatose cardiac arrest patients were evaluated with brain CT. Of these patients, after exclusion of patients whose brain CT scans were done too late or with poor baseline neurology (Cerebral Performance Category [CPC] ≥ 3), 91 patients were included for this study. The parameters of initial brain CT, i.e., gray matter-to-white matter ratio (GWR) and ONSD, were measured after clinical care as part of a retrospective reanalysis of images. ONSD on brain CT was bilaterally measured 3 mm behind the eyeball at fixed window width and level and averaged to yield the mean value. The performance of ONSD to predict poor neurologic outcome (CPC = 3 to 5) was analyzed using multiple logistic regression analysis, receiver operating characteristic (ROC) curve analysis, and cross-tabulations. RESULTS: Twenty-three patients showed good neurologic outcomes at hospital discharge. Mean (±SD) ONSD was 5.6 (±0.3) mm in the good outcome group versus 6.3 (±0.5) mm in the poor outcome group (p < 0.001). After basic clinical covariates were controlled for, i.e., age, Glasgow Coma Scale (GCS) score (3 vs. 4-15), and time from collapse to return of spontaneous circulation (ROSC), ONSD (odds ratio [OR] = 2.1; 95% confidence interval [CI] = 1.1 to 3.9) and GWR (OR = 0.6; 95% CI = 0.4 to 0.9) were found to be significant factors for predicting poor neurologic outcome. ROC curve analysis showed that ONSD and GWR had areas under the ROC curve of 0.931 (95% CI = 0.87 to 0.98) and 0.922 (95% CI = 0.86 to 0.97), respectively. Combining the cutoff values of ONSD (6.21 mm, sensitivity = 56%; 95% CI = 43% to 68%) and GWR (1.23, sensitivity = 84%; 95% CI = 73% to 92%) to have 100% specificities, the sensitivity was improved to 92% (95% CI = 84% to 98%). Intrarater and interrater intraclass correlation coefficients between the investigators measuring ONSD were 0.888 and 0.833, respectively. CONCLUSIONS: Optic nerve sheath diameter on initial brain CT correlated closely with the neurologic outcome of hypoxic ischemic encephalopathy and had good reliability. Additional prospective work may be justified evaluating the standardization and diagnostic performance in real time use as a predictive tool for neurologic outcome following cardiac arrest.

Ionic liquid-mediated extraction of lipids from algal biomass.

Lipids from algal biomass were extracted using mixtures of ionic liquids (ILs) and methanol, and fatty acid profiles of the extracted lipids were characterized in this work. Mixtures of ILs and methanol successfully dissolved biomass leaving lipids insoluble. The total contents of lipids extracted from commercial and cultivated Chlorella vulgaris were 10.6% and 11.1%, respectively, by the conventional Bligh and Dyer's method, while a mixture of [Bmim][CF(3)SO(3)] and methanol extracted 12.5% and 19.0% of the lipids, respectively. Multi-parameter regression by the linear solvation energy relationship showed that dipolarity/polarizability and hydrogen bond acidity of ILs are more important than their hydrogen bond basicity for effectively extracting lipids from algal biomass. Fatty acid profiles of the lipids extracted using IL-methanol mixtures showed that C16:0, C16:1, C18:2, and C18:3 fatty acids were dominant. This suggests that the lipids extracted from C. vulgaris can be used as a source of biodiesel production.