Cold Ethanol Extraction of Cannabinoids and Terpenes from Cannabis Using Response Surface Methodology: Optimization and Comparative Study.

Efficient cannabis biomass extraction can increase yield while reducing costs and minimizing waste. Cold ethanol extraction was evaluated to maximize yield and concentrations of cannabinoids and terpenes at different temperatures. Central composite rotatable design was used to optimize two independent factors: sample-to-solvent ratio (1:2.9 to 1:17.1) and extraction time (5.7 min-34.1 min). With response surface methodology, predicted optimal conditions at different extraction temperatures were a cannabis-to-ethanol ratio of 1:15 and a 10 min extraction time. With these conditions, yields (g 100 g dry matter-1) were 18.2, 19.7, and 18.5 for -20 °C, -40 °C and room temperature, respectively. Compared to the reference ground sample, tetrahydrocannabinolic acid changed from 17.9 (g 100 g dry matter-1) to 15, 17.5, and 18.3 with an extraction efficiency of 83.6%, 97.7%, 102.1% for -20 °C, -40 °C, and room temperature, respectively. Terpene content decreased by 54.1% and 32.2% for extraction at -20 °C and room temperature, respectively, compared to extraction at -40 °C. Principal component analysis showed that principal component 1 and principal component 2 account for 88% and 7.31% of total variance, respectively, although no significant differences in cold ethanol extraction at different temperatures were observed.

Microwave- and Ultrasound-Assisted Extraction of Cannabinoids and Terpenes from Cannabis Using Response Surface Methodology.

Limited studies have explored different extraction techniques that improve cannabis extraction with scale-up potential. Ultrasound-assisted and microwave-assisted extraction were evaluated to maximize the yield and concentration of cannabinoids and terpenes. A central composite rotatable design was used to optimize independent factors (sample-to-solvent ratio, extraction time, extraction temperature, and duty cycle). The optimal conditions for ultrasound- and microwave-assisted extraction were the sample-to-solvent ratios of 1:15 and 1:14.4, respectively, for 30 min at 60 °C. Ultrasound-assisted extraction yielded 14.4% and 14.2% more oil and terpenes, respectively, compared with microwave-assisted extracts. Ultrasound-assisted extraction increased cannabinoid concentration from 13.2−39.2%. Considering reference ground samples, tetrahydrocannabinolic acid increased from 17.9 (g 100 g dry matter−1) to 28.5 and 20 with extraction efficiencies of 159.2% and 111.4% for ultrasound-assisted and microwave-assisted extraction, respectively. Principal component analyses indicate that the first two principal components accounted for 96.6% of the total variance (PC1 = 93.2% and PC2 = 3.4%) for ultrasound-assisted extraction and 92.4% of the total variance (PC1 = 85.4% and PC2 = 7%) for microwave-assisted extraction. Sample-to-solvent ratios significantly (p < 0.05) influenced the secondary metabolite profiles and yields for ultrasound-assisted extracts, but not microwave-assisted extracts.

Factors Influencing the Dissemination of Tweets at the American College of Surgeons Clinical Congress 2018.

BACKGROUND: Social media is increasingly used in surgery, facilitating the dissemination of knowledge. Social media can potentially aid networking, education, and information exchange. This study explored the impact of tweet components and tweeter characteristics during a large surgical congress to inform recommendations for optimizing social media use at future surgical conferences. METHODS: Twitter activity was monitored during the American College of Surgeons Clinical Congress 2018 using NodeXL to extract tweets containing the conference hashtag #ACSCC18 (or #ACSCC2018). Multivariable logistic regression analysis was performed to identify independent predictors of retweet activity, also testing for multicollinearity and interactions among variables. RESULTS: There were 4386 tweets that contained #ACSCC18 (or #ACSCC2018) posted from 1023 Twitter accounts. A larger group of Twitter accounts just retweeted. Other Twitter accounts with a stake in the conference neither tweeted nor retweeted. In a multivariable analysis of original tweets, the following were all independently associated with retweets, in decreasing order of effect size: mention of other tweeters, multimedia, inclusion of other hashtags, and the number of followers. In contrast with other conferences, the inclusion of a weblink (URL)-for example, link to paper or blog-was not associated with retweets. DISCUSSION: This study helps understand social media impact at surgical conferences. Engage by tweeting and retweeting. Mention other tweeters, add multimedia, include congress hashtags and topic-specific hashtags, and build your followers. Although not associated with retweet activity in this study, the inclusion of URLs can still contribute in substantiating the disseminated content based on findings at other conferences.

World Antibiotic Awareness Week and European Antibiotic Awareness Day, November 2018: An analysis of the impact of Twitter activity.

INTRODUCTION: International antibiotic awareness and stewardship campaigns use social media to disseminate campaign materials. Analysis of campaigns during 2016 and 2017 identified a wide range of hashtags in use, potentially leading to confusion and dilution of the campaign message. This study aimed to examine the impact of different hashtags on the dissemination of information during European Antibiotic Awareness Day and World Antibiotic Awareness Week, November 2018. METHODS: Tweets were prospectively extracted using Followthehashtag and analysed in Excel and R (14 400 tweets by 5 899 tweeters, with 60 222 retweets). Descriptive analysis and multivariable logistic regression analyses were adjusted for potential confounders. RESULTS: There was a positive association between retweets received and inclusion of media, hashtag(s), mention of other tweeter(s), and number of followers. There was hashtag drift (e.g. the unofficial #WAAW2018 hashtag was used more frequently than the official campaign #WAAW18 hashtag). Analysis of five popular hashtags from the European and worldwide campaigns found a positive association between retweets and the unofficial hashtag #WAAW2018 and the long-established #AntibioticGuardian hashtag. #AntibioticResistance (another official hashtag) was the most frequently used hashtag but did not have a consistent positive association with retweets. Analysis of the contribution of tweeters - tweeters, retweeters, and mentions of tweeters - demonstrated 32 534 accounts in total, 95% of which tweeted and/or retweeted, the great majority being retweeters. CONCLUSION: This study demonstrates the breadth of participation in international antibiotic awareness campaigns, but also the need for clarity about campaign hashtags, and the need to search beyond official hashtags when evaluating the impact of a campaign.

DNMT3B Oncogenic Activity in Human Intestinal Cancer Is Not Linked to CIMP or BRAFV600E Mutation.

Approximately 10% of human colorectal cancer (CRC) are associated with activated BRAFV600E mutation, typically in absence of APC mutation and often associated with a CpG island methylator (CIMP) phenotype. To protect from cancer, normal intestinal epithelial cells respond to oncogenic BRAFV600E by activation of intrinsic p53 and p16-dependent tumor suppressor mechanisms, such as cellular senescence. Conversely, CIMP is thought to contribute to bypass of these tumor suppressor mechanisms, e.g. via epigenetic silencing of tumor suppressor genes, such as p16. It has been repeatedly proposed that DNMT3B is responsible for BRAFV600E-induced CIMP in human CRC. Here we set out to test this by in silico, in vitro, and in vivo approaches. We conclude that although both BRAFV600E and DNMT3B harbor oncogenic potential in vitro and in vivo and show some evidence of cooperation in tumor promotion, they do not frequently cooperate to promote CIMP and human intestinal cancer.

Early Cannulation ePTFE Arteriovenous Access Grafts are Associated with a Low Incidence of Pseudoaneurysm Formation.

BACKGROUND: Pseudoaneurysm formation is common in standard thin-walled polytetrafluoroethylene (sPTFE) grafts, occurring in up to 10% of grafts, and is reported as the most common cause of graft loss for grafts more than 2 years old. The Gore® Acuseal™ graft is an early cannulation graft, needled before incorporation, and thus may be especially prone to pseudoaneurysm formation. In addition, as this is a relatively new product, there are limited data on long-term outcomes such as pseudoaneurysm. We report one center's experience of the incidence and etiological factors associated with pseudoaneurysm formation over 5 years and 265 grafts. METHODS: A total of 265 Acuseal grafts were placed in the last 5 years. All patients had prospective data entered into an electronic searchable patient record. Surveillance was performed with 3 monthly imaging (digital subtraction angiography or ultrasound), clinical examination, and hemodynamic performance. Data examined included the incidence, causative factors, and outcomes of pseudoaneurysm. RESULTS: Eleven grafts (4.15%) developed a pseudoaneurysm, with 2 patients developing significant hemorrhage. The median time to development of a pseudoaneurysm was 25 months interquartile range (IQR, 20-28 months). Several common etiological factors were identified. All but one patient had overuse of needling sites (n = 10; 90.9%). Other factors associated with pseudoaneurysm formation were inadequate surveillance (n = 9; 81.8%), venous outflow stenosis (n = 9; 81.8%), and anticoagulation/dual antiplatelet therapy (n = 7; 63.6%). Management included observation and needle rotation (n = 5; 45.5%), stent grafting (n = 3; 27.3%), or excision (n = 1; 9.1%) of the pseudoaneurysm. Surgical or endovascular augmentation of the venous outflow was required in 9 patients (81.8%). Graft ligation and explantation were required in 5 patients (45.5%) with graft preservation achieved in 6/11 patients (54.5%). CONCLUSIONS: Pseudoaneurysm formation occurs less frequently in Acuseal grafts compared with historical data for standard PTFE grafts. Pseudoaneurysm formation did not occur in any graft within the first 13 months after implantation, suggesting early cannulation before incorporation is not by itself a risk factor for pseudoaneurysm development. Poor needling, venous stenosis, inadequate surveillance, and anticoagulation/dual antiplatelet therapy are remediable factors, and graft preservation is possible. Acuseal is a robust graft with lower rates of pseudoaneurysm formation on long-term follow-up than standard PTFE grafts.

Improving the quality and impact of public health social media activity in Scotland during 2016: #ScotPublicHealth.

Background: Social media, including Twitter, potentially provides a route to communicate public health messages to a large audience. Simple measures can boost onward broadcast to other users ('retweeting'). This study compares the impact of a structured programme of social media activity in Scotland during 2016 (using #ScotPublicHealth hashtag) with previous years. Methods: The Twitter search function was used to identify tweets between 2014 and 2016 inclusive. The first three tweets from each Twitter user were selected for each period. The number of retweets was used as a measure of impact. The quality of tweets was assessed by recording use of image, weblink (uniform resource locator or URL), mention of another Twitter user and/or hashtag, each of which have been shown to boost number of retweets. Results: The percentage of tweets with an image, URL and/or mention of another Twitter user increased during the period of study. The percentage of tweets retweeted during Scottish Public Health conferences increased from 43% in 2014 to 70% in 2016. The volume of tweeting also increased. Conclusions: The quality and impact of tweets sent by the Scottish Public Health community was higher during 2016 than previous years. Conference tweeting remains an area for improvement.

Oncogene-Expressing Senescent Melanocytes Up-Regulate MHC Class II, a Candidate Melanoma Suppressor Function.

On acquisition of an oncogenic mutation, primary human and mouse cells can enter oncogene-induced senescence (OIS). OIS is characterized by a stable proliferation arrest and secretion of proinflammatory cytokines and chemokines, the senescence-associated secretory phenotype. Proliferation arrest and the senescence-associated secretory phenotype collaborate to enact tumor suppression, the former by blocking cell proliferation and the latter by recruiting immune cells to clear damaged cells. However, the interactions of OIS cells with the immune system are still poorly defined. Here, we show that engagement of OIS in primary human melanocytes, specifically by melanoma driver mutations NRASQ61K and BRAFV600E, causes expression of the major histocompatibility class II antigen presentation apparatus, via secreted IL-1ß signaling and expression of CIITA, a master regulator of major histocompatibility class II gene transcription. In vitro, OIS melanocytes activate T-cell proliferation. In vivo, nonproliferating oncogene-expressing melanocytes localize to skin-draining lymph nodes, where they induce T-cell proliferation and an antigen presentation gene expression signature. In patients, expression of major histocompatibility class II in melanoma is linked to favorable disease outcome. We propose that OIS in melanocytes is accompanied by an antigen presentation phenotype, likely to promote tumor suppression via activation of the adaptive immune system.

Dual Strain-Promoted Alkyne-Nitrone Cycloadditions for Simultaneous Labeling of Bacterial Peptidoglycans.

Bioorthogonal chemistry has been applied to study a multitude of biological processes in complex environments through incorporation and detection of small functional groups. However, few reactions are known to be compatible with each other to allow for studies of more than one biomolecule simultaneously. Here we describe a dual labeling method wherein two stereoelectronically contrasting nitrone tags are incorporated into bacteria peptidoglycan and detected via strain-promoted alkyne-nitrone cycloaddition (SPANC) simultaneously. Furthermore, we show orthogonality with the azide functionality broadening the potential for simultaneous biomolecular target labeling in less accommodating metabolic pathways. We also demonstrate the simultaneous labeling of two different food-associated bacteria, L. innocua (a model for the food-born pathogen L. monocytogenes) and L. lactis (a fermentation bacterium). The ability to monitor multiple processes and even multiple organisms concurrently through nitrone/nitrone or nitrone/azide incorporation strengthens the current bioorthogonal toolbox and gives rise to robust duplex labeling of organisms to potentiate the studies of rapid biological phenomena.

Bioorthogonal labelling of living bacteria using unnatural amino acids containing nitrones and a nitrone derivative of vancomycin.

Unnatural D-amino acids bearing endocyclic nitrones were developed for live-cell labelling of the bacterial peptidoglycan layer. Metabolic incorporation of D-Lys and D-Ala derivatives bearing different endocyclic nitrones was observed in E. coli, L. innocua, and L. lactis. The incorporated nitrones of these bacteria then rapidly underwent strain-promoted alkyne-nitrone cycloaddition (SPANC) reactions affording chemically modified bacteria.

Correction: Chigrinova, M., et al. Kinugasa reactions in water: from green chemistry to bioorthogonal labelling. Molecules 2015, 20, 6959-6969.

The authors wish to make the following correction to this paper [1]: The author name "Paul Pezacki" should be "John Paul Pezacki". [...].

Kinugasa reactions in water: from green chemistry to bioorthogonal labelling.

The Kinugasa reaction has become an efficient method for the direct synthesis of ß-lactams from substituted nitrones and copper(I) acetylides. In recent years, the reaction scope has been expanded to include the use of water as the solvent, and with micelle-promoted [3+2] cycloadditions followed by rearrangement furnishing high yields of ß-lactams. The high yields of stable products under aqueous conditions render the modified Kinugasa reaction amenable to metabolic labelling and bioorthogonal applications. Herein, the development of methods for use of the Kinugasa reaction in aqueous media is reviewed, with emphasis on its potential use as a bioorthogonal coupling strategy.

Strain-promoted cycloadditions involving nitrones and alkynes--rapid tunable reactions for bioorthogonal labeling.

The development and applications of strain-promoted alkyne-nitrone cycloaddition (SPANC) reactions have brought about new tools for rapid and specific functionalization of biomolecules in different settings. While a number of strain-promoted reactions have been successfully developed, SPANC reactions offer high reactivity with bimolecular rate constants of k2 that are as fast as 60M(-1)s(-1). SPANC reactions also offer stability of starting materials, particularly in the case of endocyclic nitrones, as well as stereoelectronic tunability of the nitrone moiety to optimize reactivity towards different alkyne reaction partners. Herein we discuss recent advances in the development of SPANC reactions and their applications in bioorthogonal labeling.

Suction-based grasping tool for removal of regular- and irregular-shaped intraocular foreign bodies.

BACKGROUND AND OBJECTIVE: To describe a suction-based grasping tool for the surgical removal of irregular-shaped and nonferromagnetic intraocular foreign bodies. MATERIALS AND METHODS: A surgical tool with suction capabilities, consisting of a stainless steel shaft with a plastic handle and a customizable and interchangeable suction tip, was designed in order to better engage and manipulate irregular-shaped in-traocular foreign bodies of various sizes and physical properties. The maximal suction force and surgical capabilities were assessed in the laboratory and on a cadaveric eye vitrectomy model. RESULTS: The suction force of the water-tight seal between the intraocular foreign body and the suction tip was estimated to be approximately 40 MN. During an open-sky vitrectomy in a porcine model, the device was successful in engaging and firmly securing foreign bodies of different sizes and shapes. CONCLUSION: The suction-based grasping tool enables removal of irregular-shaped and nonferromagnetic foreign bodies.

Management of a dislocated intraocular lens with a suction-based grasping tool.

UNLABELLED: Spontaneous late subluxation of an intraocular lens (IOL) is a rare but serious complication of cataract surgery, often associated with poor zonular support. The management of this complication with a conventional forceps is difficult because the forceps often cannot adequately grasp the optic or haptic to fixate the IOL to the sclera or iris or remove it. The new suction-based grasping tool has a suction cup at the end. The combination of a suction vector and the use of a set of specially designed suction tips allow grasping and stabilization of the subluxated IOL. The properties of the suction-based grasping tool provide better control of the subluxated IOL, simplifying the management of these challenging cases. FINANCIAL DISCLOSURE: Drs. Olson and Mackenzie have a patent application pending for the disclosed device. No other author has a financial or proprietary interest in any material or method mentioned.

A prospective comparison of the prognostic value of tumor- and patient-related factors in patients undergoing potentially curative surgery for pancreatic ductal adenocarcinoma.

BACKGROUND: Outcome prediction after resection with curative intent for pancreatic ductal adenocarcinoma remains a challenge. There is increasing evidence that the presence of an ongoing systemic inflammatory response is associated with poor outcome in patients undergoing resection for a variety of common solid tumors. Our aim was to prospectively evaluate the prognostic value of tumor- and patient-related factors including the systemic inflammatory response in patients undergoing potentially curative surgery for pancreatic ductal adenocarcinoma of the head of pancreas. METHODS: The prognostic impact of tumor factors such as stage and host factors, including the systemic inflammatory response (modified Glasgow Prognostic Score [mGPS]), were evaluated in a prospective study of 135 patients who underwent elective pancreaticoduodenectomy for pancreatic ductal adenocarcinoma from January 2002 to April 2009. RESULTS: In addition to the established tumor-related pathological factors (in particular margin involvement; hazard ratio [HR] 2.82, 95% confidence interval [CI] 1.65-4.84, P < 0.001), an elevated mGPS (HR 2.26, 95% CI 1.43-3.57, P < 0.001) was independently associated with lower overall survival after pancreaticoduodenectomy. Additionally, in an adjuvant therapy subgroup of 74 patients, both margin involvement and an elevated mGPS remained independently associated with reduced overall survival. CONCLUSIONS: We have prospectively validated the influence of tumor-related and patient-related factors. Margin involvement and the preoperative mGPS were the most important determinants of overall survival in patients undergoing potentially curative pancreaticoduodenectomy. Furthermore, both had independent prognostic value in those patients receiving adjuvant chemotherapy. In the future, this may be considered a stratification factor for entry onto therapeutic trials.

Circulating methylarginine levels and the decline in renal function in patients with chronic kidney disease are modulated by DDAH1 polymorphisms.

In patients with chronic kidney disease, high plasma levels of the endogenous nitric oxide synthase inhibitor, asymmetric dimethylarginine, are thought to contribute to decline in renal function. Here we took a candidate gene approach to determine any causal role of asymmetric dimethylarginine in the progression of chronic kidney disease. The impact of single-nucleotide polymorphisms in the genes encoding the two isoforms of the asymmetric dimethylarginine-degrading enzyme, dimethylarginine dimethylaminohydrolase (DDAH1 and DDAH2), on enzyme expression, plasma asymmetric dimethylarginine levels, and longitudinal changes in estimated glomerular filtration rate were determined in various patient groups. There was evidence suggesting that the rs17384213 DDAH1 GG genotype was associated with increased expression of its mRNA in kidney allografts. Healthy subjects carrying the rs17384213 G allele had lower plasma asymmetric dimethylarginine, and a similar borderline association was found in patients with chronic kidney disease. This allele, however, was independently associated with a steeper decline in renal function in two separate cohorts of patients with chronic kidney disease. We conclude that polymorphisms in DDAH1 alter the rate of decline of glomerular filtration rate in subjects with chronic kidney disease. Our findings show that DDAH1 modulates plasma asymmetric dimethylarginine and contributes to the decline in renal function. However, it appears that increases in circulating methylarginine did not mediate progressive kidney injury.

Suprasegmental characteristics of speech produced during simultaneous communication by inexperienced signers.

This study investigated suprasegmental variables of syllable stress and intonation contours in contextual speech produced during simultaneous communication (SC) by inexperienced signers. Ten hearing inexperienced sign language users were recorded under SC and speech-alone (SA) conditions speaking a set of sentences containing stressed versus unstressed versions of the same syllables and a set of sentences containing interrogative versus declarative versions of the same words. Results indicated longer sentence durations for SC than SA for all speech materials. Vowel duration and fundamental frequency differences between stressed and unstressed syllables as well as intonation contour differences between declarative and interrogative sentences were essentially the same in both SC and SA conditions. The conclusion that prosodic rules were not violated by inexperienced signers in SC is consistent with previous research indicating that temporal alterations produced during SC do not involve degradation of other temporal or spectral characteristics of English speech.

Mumps in a boarding school: description of an outbreak and control measures.

BACKGROUND: The number of cases of mumps in the UK has increased during 2004-2005. Understanding why some people are more susceptible to mumps infection will help target vaccination and other control measures in the future. This paper describes a mumps outbreak in a boarding school in Scotland (October to November 2004). OBJECTIVES: To describe the characteristics of cases in a mumps outbreak; and to conduct a case-control study looking at the effect of previous measles-mumps-rubella (MMR) vaccination. DESIGN: Descriptive epidemiology. DATA SOURCES: Clinical, laboratory and surveillance data. SETTING: A private school with 600 boarding and day pupils. RESULTS: Fifty cases were notified to the public health department as having mumps. Twenty of the cases (40%) were confirmed virologically, all born between 1987-1991. A matched case-control study was conducted to explore the effect of prior MMR vaccination. Compared to no vaccine, MMR protected against mumps (odds ratio = 0.7), two doses offering best protection (odds ratio = 0.5), but the study was not large enough to detect a statistically significant difference. CONCLUSION: Some children and young people in the UK are currently incompletely vaccinated. This outbreak illustrates the potential of a mumps outbreak to disrupt the medical, educational and social life of a school. All children should be vaccinated with two doses of MMR vaccine before school entry. Children and young people born between 1979-1991 should be given the necessary additional dose(s) of MMR vaccine.