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The arachidonic acid derivatives N-arachidonoylethanolamine (anandamide; AEA), 2-arachidonoylglycerol (2-AG), N-arachidonoyldopamine (NADA), 2-arachidonoylglycerol ether (noladin ether; 2-AGE) and O-arachidonoylethanolamine (virodhamine; VA) were identified as physiological components of the endocannabinoid (EC) system. In order to gain further profound knowledge about the different EC-induced physiological and pathophysiological effects, appropriate analytical methods are required. The method described here uses liquid chromatography in combination with positive electrospray ionization mass spectrometry (LC-MS/MS) to quantify the concentrations of the above-mentioned EC compounds in cells. Sample preparation prior to LC-MS/MS analysis was performed by means of two liquid extractions with ethyl acetate. The method has been validated according to the bioanalytical guidelines of the Food and Drug Administration (FDA). The lower limits of quantification were 0.03 ng/mL for AEA, 2 ng/mL for 2-AG, 0.03 ng/mL for NADA, 0.3 ng/mL for 2-AGE and 0.15 ng/mL for VA. Linearity was demonstrated up to 10 ng/mL (AEA, NADA, 2-AGE and VA) and 50 ng/mL (2-AG). The values for intra- and inter-day precision and accuracy were within the guideline recommended acceptance criteria for assay validation. Low matrix effects and good recovery were found for AEA, 2-AG and 2-AGE, while a higher matrix effect was observed for NADA and VA. Extraction yields were lowest for VA. The method was used for EC measurement in different cell lines and in mouse brains.Human fertilization success depends on the ability of the spermatozoa to undergo capacitation. Even though this process can be conducted in vitro, the optimal time for a sperm cell to complete capacitation in vitro is still under discussion due to the lack of proper capacitation biomarkers. Here, we evaluated the influence of in vitro capacitation time on HSPA2 distribution over human sperm head testing this chaperone as a potential capacitation biomarker. The chaperone was assessed in human spermatozoa from 16 normozoospermic donors using indirect immunofluorescence in uncapacitated, one and four-hour capacitated spermatozoa. The percentage of HSPA2 immunofluorescent cells before and after one hour of capacitation did not differ significantly. However, after four hours of capacitation, we observed a significantly higher percentage of HSPA2 labelled cells. In fluorescent cells analysed before capacitation, we could not identify a predominant distribution pattern. Meanwhile, after capacitation, most sperm showed a highly labelled equatorial band accompanied by a homogeneous fluorescence throughout the acrosomal region. Our findings suggest that HSPA2 needs more than one hour of in vitro capacitation for being correctly distributed in the anterior region of the sperm head. Selleckchem RMC-6236 In conclusion, the present study provides solid evidences for the utility of HSPA2 as a biomarker of human sperm in vitro capacitation. Due to its importance during egg-sperm recognition, the use of HSPA2 as a biomarker before an artificial reproduction technique may be suggested, in addition to a longer capacitation time during sperm preparation.
Evaluate whether intraprocedural MRI monitoring of percutaneous cryoablation procedures of head and neck, and spine lesions is effective for avoiding iatrogenic neurovascular and mucosal injury.
We retrospectively reviewed 64 consecutive percutaneous head and neck, and spine cryoablation procedures with intraprocedural MRI monitoring performed on 45 patients (mean age 55years, range 17-91years). Ablation goals were either complete local control of primary or metastatic lesions or pain relief.
The technical success rate was 100%. The complication rate was 13% with only 2 complications (3%) requiring further intervention. There were no deaths or persistent neurological or vascular complications. Subsequent cryoablation in the same location was performed in 12 patients (27%). Subsequent surgical intervention in the same location was performed in 7 patients (16%) for progressive disease or worsening symptoms.
MRI provides excellent visualization of the ice ball margin during percutaneous cryoablation procedures. Accurate intraprocedural visualization of the ice ball allows for adjustment of cryoablation parameters to avoid damage to adjacent vital neurovascular structures or mucosal surfaces. Intraprocedural MRI monitoring is thus a novel and highly effective method that allows a high rate of technical success for cryoablation in the head and neck, and spine while avoiding iatrogenic injury.
MRI provides excellent visualization of the ice ball margin during percutaneous cryoablation procedures. Accurate intraprocedural visualization of the ice ball allows for adjustment of cryoablation parameters to avoid damage to adjacent vital neurovascular structures or mucosal surfaces. Intraprocedural MRI monitoring is thus a novel and highly effective method that allows a high rate of technical success for cryoablation in the head and neck, and spine while avoiding iatrogenic injury.
The aim of our study was to evaluate the yield of head CT in the ED in different age groups and different referral indications.
Records of one large academic tertiary care ED were retrospectively reviewed for consecutive adult patients who underwent a head CT between January 1st 2017 and February 10th 2017. CT referral forms and interpretations were obtained and evaluated for demographics, referral indications, and findings. Scans were divided into three groups acute findings, chronic findings, and normal. The cohort was divided into three age groups. Associations between referral indications and acute findings were calculated.
Overall, 1536 of adult patients with ED head CT were included. Acute findings were found in 239/1536 (15.5%) of the CTs. The frequency of acute findings increased with age (p=0.027). The most common acute findings were brain hemorrhage (32.6%), infarct (27.6%), and mass (23%). The top three referral indications were focal neurologic deficit (28%), trauma (24.7%), and headache (17.5%). The rates of positive acute findings for different referral indications were seizure 27%, confusion 20%, syncope 19%, focal neurologic deficit 16%, head injury 15%, headache 12%, and dizziness 8%.
This study shows the yield of ED head CT for acute findings for different age groups and for different referral indications. The frequency of acute findings increased with age. Suspected seizure had the highest association with an acute finding, whereas dizziness had the lowest association.
This study shows the yield of ED head CT for acute findings for different age groups and for different referral indications. The frequency of acute findings increased with age. Suspected seizure had the highest association with an acute finding, whereas dizziness had the lowest association.
Homepage: https://www.selleckchem.com/products/rmc-6236.html
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