"The Perfect Formula:" Evaluating Health Claims, Products and Pricing on Cannabis Dispensary Websites in Two Recently Legalized States.

Introduction: Many cannabis dispensaries market and sell their products online through websites designed to attract and maintain customers; often, these websites incorporate a variety of product claims and other marketing tactics. This study evaluated website content, product pricing and discounts on dispensary websites in California and Nevada, states that legalized recreational cannabis in 2016. Methods: We content coded product availability, marketing claims and discounts on cannabis dispensary websites in the San Francisco Bay Area (N = 34) and Reno (N = 15) from March to June 2020 using a web crawler to scrape pricing information for four product types. We conducted bivariate analyses comparing both locations. Results: Prices were significantly lower for flower, edibles, and concentrates in Reno compared to the Bay Area, but not cartridges. In both areas, a range of marketing claims were made regarding the health effects of certain products. The most common were that cannabis products treated pain, nausea/vomiting, spasms, anxiety, insomnia, and depression. Products were also said to promote creativity and euphoria. Other marketing claims related to potency, pleasure enhancement, and improved social interactions. Discounts targeted to senior citizens and veterans were found on over half of all websites. Conclusions: Dispensary websites in the Bay Area and Reno frequently make health-related claims which should not be allowed in absence of scientific evidence. Non-health related claims are similar to those used for selling e-cigarettes and other tobacco products. Monitoring cannabis dispensary websites provides insight into local sales tactics and may help identify subpopulations for research on behavioral impacts of cannabis marketing activities.

Cannabis dispensary online marketing practices in response to COVID-19 lockdowns.

Background: The COVID-19 pandemic has been accompanied by increases in cannabis consumption, which might relate to dispensary marketing activities. As part of an ongoing project monitoring cannabis dispensary websites in Northern California and Reno, Nevada, we noticed many websites added announcements and "pop-up" communications in response to lockdowns. This brief report describes the cannabis dispensary website communications related to COVID-19 with the aim to provide insight into emerging marketing messages that may increase cannabis consumption in times of crisis. Methods: Content analysis of COVID-19 announcements present on cannabis dispensary websites in San Francisco/Alameda Counties (n = 32), and the Reno area (n = 15) in April-May 2020 shortly after lockdowns were implemented. Results: COVID-19 announcements were present on 25/32 (78%) of dispensary websites in San Francisco/Alameda and 9/15 (60%) of websites in the Reno area. Almost all COVID-19 announcements (88% San Francisco/Alameda, 89% Reno) announced operational changes such as delivery or curbside pickup services, 72% and 56% respectively announced patron/employee safety measures. Health related messages were present; about half of website announcements referred to government/health authorities, 44% of Reno area announcements used healthcare rhetoric, and some San Francisco/Alameda announcements included suggestions for using cannabis to mitigate infection risk or manage anxiety. Conclusions: Most cannabis dispensaries in the study region implemented COVID-19 pandemic operational changes to maintain product availability, and many positioned their identity with health - either by referring to health authorities, or using health rhetoric, and a minority gave health advice. Cannabis dispensary websites provide a timely snapshot of marketing practices that may contribute to increases in cannabis use during stressful events.

Urinary-cell mRNA profile and acute cellular rejection in kidney allografts.

BACKGROUND: The standard test for the diagnosis of acute rejection in kidney transplants is the renal biopsy. Noninvasive tests would be preferable. METHODS: We prospectively collected 4300 urine specimens from 485 kidney-graft recipients from day 3 through month 12 after transplantation. Messenger RNA (mRNA) levels were measured in urinary cells and correlated with allograft-rejection status with the use of logistic regression. RESULTS: A three-gene signature of 18S ribosomal (rRNA)-normalized measures of CD3ε mRNA and interferon-inducible protein 10 (IP-10) mRNA, and 18S rRNA discriminated between biopsy specimens showing acute cellular rejection and those not showing rejection (area under the curve [AUC], 0.85; 95% confidence interval [CI], 0.78 to 0.91; P<0.001 by receiver-operating-characteristic curve analysis). The cross-validation estimate of the AUC was 0.83 by bootstrap resampling, and the Hosmer-Lemeshow test indicated good fit (P=0.77). In an external-validation data set, the AUC was 0.74 (95% CI, 0.61 to 0.86; P<0.001) and did not differ significantly from the AUC in our primary data set (P=0.13). The signature distinguished acute cellular rejection from acute antibody-mediated rejection and borderline rejection (AUC, 0.78; 95% CI, 0.68 to 0.89; P<0.001). It also distinguished patients who received anti-interleukin-2 receptor antibodies from those who received T-cell-depleting antibodies (P<0.001) and was diagnostic of acute cellular rejection in both groups. Urinary tract infection did not affect the signature (P=0.69). The average trajectory of the signature in repeated urine samples remained below the diagnostic threshold for acute cellular rejection in the group of patients with no rejection, but in the group with rejection, there was a sharp rise during the weeks before the biopsy showing rejection (P<0.001). CONCLUSIONS: A molecular signature of CD3ε mRNA, IP-10 mRNA, and 18S rRNA levels in urinary cells appears to be diagnostic and prognostic of acute cellular rejection in kidney allografts. (Funded by the National Institutes of Health and others.).

Hotspot Exon 15 Mutations in BRAF Are Uncommon in Feline Tumours.

BRAF is one of multiple RAF proteins responsible for the activation of the MAPK cell signalling cascade involved in cell growth, differentiation, and survival. A hotspot BRAFV600E mutation, in exon 15, was determined to be a driver in 100% hairy cell leukaemias, 50%-60% of human melanomas, 30%-50% of human thyroid carcinomas and 10%-20% of human colorectal carcinomas. The orthologous BRAFV595E mutation was seen in 67% and 80% of canine bladder transitional cell carcinomas and prostatic adenocarcinomas, respectively. Since veterinary and human cancers exploit similar pathways and BRAF is highly conserved across species, BRAF can be expected to be a driver in a feline cancer. Primers were developed to amplify exon 15 of feline BRAF. One hundred ninety-six feline tumours were analysed. Sanger sequencing of the 211 bp PCR amplicon was done. A BRAF mutation was found in one tumour, a cutaneous melanoma. The mutation was a BRAFV597E mutation, orthologous to the canine and human hotspot mutations. A common synonymous variant, BRAFT586T, was seen in 23% (47/196) of tumours. This variant was suspected to be a single nucleotide polymorphism. BRAF was not frequently mutated in common feline tumours or in tumour types that frequently harbour BRAF mutations in human and canine cancers. As is seen in canine cancer genomics, the mutational profile in feline tumours may not parallel the histologic equivalent in human oncology.

COVID-19-Associated Coagulopathy in the Peripartum Setting: A Case Report.

Sepsis-induced coagulopathy (SIC) scoring and D-dimer can be used to recognize COVID-19-induced coagulopathy, but the utility of these is largely unknown in the peripartum setting and leaves anticoagulation guidance unclear. We present the case of a critically ill postpartum patient with COVID-19 infection. This patient presented with clinical signs of COVID-19 infection and developed acute respiratory failure requiring invasive mechanical ventilation and subsequent cesarean delivery at 34 weeks. She initially improved postoperatively but deteriorated after postoperative day 5. She was found to have a very elevated D-dimer of 58 µg/mL and anticoagulation was escalated to full dosing. She required prolonged mechanical ventilation and deceased after developing gram-positive cocci bacteremia. This case demonstrates that recognition and management of COVID-19-associated coagulopathy can be confusing in the peripartum period and studies are needed to validate D-dimer and SIC scoring in this population of patients.

Combining radiation therapy with zoledronate for the treatment of osteo-invasive feline oral squamous cell carcinoma.

Feline oral squamous cell carcinoma (FOSCC) is the most common oral tumour diagnosed in pet cats and carries a poor prognosis with <10% one-year survival despite multi-modal therapies. Tumours of the mandible or maxilla are frequently osteo-invasive and pain can result from osteolysis. Zoledronate is a bisphosphonate that inhibits osteoclasts and reduces bone resorption. Radiation therapy (RT) is used to treat FOSCC due to anti-cancer activity and ability to improve quality of life. We hypothesized RT can be safely combined with zoledronate, and that this combinatory therapy would be efficacious, well tolerated, and result in decreased bone resorption in cats with FOSCC. SCCF1 cell line was treated with zoledronate before, concurrently, or after RT, and clonogenic assays were performed to determine if an optimal dosing schedule would be identified. Nine cats with osteoinvasive FOSCC were recruited for treatment with 4 weekly doses of 8 Gy RT combined with zoledronate administered at the first and fourth treatments. Serial CT scans were performed to assess tumour response. Safety and tolerability were monitored with hematologic and biochemical parameters, and acute radiation effects were characterized. Serum c-telopeptide (CTx) and relative bone mineral density (rBMD) by dual -energy X-ray absorptiometry (DEXA) quantified bone resorption. In vitro studies showed no clear benefit to timing of zoledronate with RT, therefore all zoledronate was administered concurrently with RT in FOSCC patients. Based on tumour volume, 4/9 (44.4%) cats achieved partial remission, 4/9 (44.4%) stable disease and 1/9 (11.1%) had progressive disease. The combinatory therapy was well-tolerated based on biochemical measurements, and all patients experienced decreased serum CTx. Combining RT with zoledronate in tumour-bearing cats is safe, well-tolerated, results in a partial remission rate of up to 44%, and decreases serum CTx, a marker of bone resorption.

Three-dimensional hydrogel culture systems support growth and determination of chemosensitivity of feline sarcoma and carcinoma cell lines.

OBJECTIVE: To evaluate feline injection site-associated sarcoma (FISAS) and oral squamous cell carcinoma (FOSCC) cells in 3-D hydrogel-based cell cultures to determine chemosensitivity to carboplatin at concentrations comparable to those eluted from carboplatin-impregnated calcium sulfate hemihydrate (C-ICSH) beads. SAMPLE: 2 immortalized cell lines, each from a histologically confirmed primary FISAS and FOSCC. PROCEDURES: Hydrogels (10% wt/vol) were formed via UV exposure from methacrylamide-functionalized gelatin dissolved in PBSS. For each cell line, approximately 100,000 cells were encapsulated per hydrogel. Three cell-seeded 3-D hydrogels were evaluated for each carboplatin concentration (0, 150, 300, 450, and 600 µM) across 3 experiments. Drug efficacy was assessed by luminescence assay 72 hours after treatment. Growth of tumor cells treated with 300 µM or 600 µM carboplatin was evaluated using live-cell morphology imaging and confocal microscopy at 3, 7, and 14 days after treatment. RESULTS: Mean half-maximal inhibitory concentration (IC50) values for FISAS and FOSCC cells ranged from 123 to 171 µM and 155 to 190 µM, respectively, based on luminescence assay. Viability at 3, 7, and 14 days for both cell lines at 300 µM carboplatin was 50%, 25%, and 5% and at 600 µM carboplatin was 25%, 10%, and < 5%. CLINICAL RELEVANCE: 3-D hydrogel cell culture systems supported growth of feline tumor cells for determination of in vitro chemosensitivity. IC50s of each cell line were within the range of carboplatin concentrations eluted from C-ICSH beads. Cells from FISAS and FOSCC cell lines treated with carboplatin showed dose-dependent and time-dependent decreases in viability.

Three-dimensional hydrogel culture systems support growth and determination of chemosensitivity of feline sarcoma and carcinoma cell lines.

OBJECTIVE: To evaluate feline injection site-associated sarcoma (FISAS) and oral squamous cell carcinoma (FOSCC) cells in 3-D hydrogel-based cell cultures to determine chemosensitivity to carboplatin at concentrations comparable to those eluted from carboplatin-impregnated calcium sulfate hemihydrate (C-ICSH) beads. SAMPLE: 2 immortalized cell lines, each from a histologically confirmed primary FISAS and FOSCC. PROCEDURES: Hydrogels (10% wt/vol) were formed via UV exposure from methacrylamide-functionalized gelatin dissolved in PBSS. For each cell line, approximately 100,000 cells were encapsulated per hydrogel. Three cell-seeded 3-D hydrogels were evaluated for each carboplatin concentration (0, 150, 300, 450, and 600 µM) across 3 experiments. Drug efficacy was assessed by luminescence assay 72 hours after treatment. Growth of tumor cells treated with 300 µM or 600 µM carboplatin was evaluated using live-cell morphology imaging and confocal microscopy at 3, 7, and 14 days after treatment. RESULTS: Mean half-maximal inhibitory concentration (IC50) values for FISAS and FOSCC cells ranged from 123 to 171 µM and 155 to 190 µM, respectively, based on luminescence assay. Viability at 3, 7, and 14 days for both cell lines at 300 µM carboplatin was 50%, 25%, and 5% and at 600 µM carboplatin was 25%, 10%, and < 5%. CLINICAL RELEVANCE: 3-D hydrogel cell culture systems supported growth of feline tumor cells for determination of in vitro chemosensitivity. IC50s of each cell line were within the range of carboplatin concentrations eluted from C-ICSH beads. Cells from FISAS and FOSCC cell lines treated with carboplatin showed dose-dependent and time-dependent decreases in viability.

Rabies exposures and pre-exposure vaccination practices among individuals with an increased risk of rabies exposure in the United States.

OBJECTIVE To identify knowledge and practices related to rabies vaccination and serologic monitoring among animal care workers in the United States. DESIGN Cross-sectional survey. SAMPLE 2,334 animal care workers (ie, veterinarians, veterinary technicians, animal control workers, and wildlife rehabilitators). PROCEDURES Participants were contacted through relevant professional organizations to participate in an anonymous web-based survey. The survey collected demographic and occupational information, animal handling and potential rabies exposure information, and individual rabies vaccination and serologic monitoring practices. Comparisons of animal bite and rabies exposure rates were made between occupational groups. Multiple logistic regression was used to evaluate factors associated with rabies vaccination status and adherence to serologic monitoring recommendations. RESULTS Respondents reported 0.77 animal bites/person-year or 0.10 bites/1,000 animals handled. The overall rate of postexposure prophylaxis due to an occupational rabies exposure was 1.07/100 person-years. Veterinarians reported the highest rabies vaccination rate (98.7% [367/372]), followed by animal control workers (78.5% [344/438]), wildlife rehabilitators (78.2% [122/156]), and veterinary technicians (69.3% [937/1,352]). Respondents working for employers requiring rabies vaccination and serologic monitoring were 32.16 and 6.14 times, respectively, as likely to be vaccinated or have a current serologic monitoring status as were respondents working for employers without such policies. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that, given the high reported rates of animal bites and potential rabies exposures among animal care workers, improvements in rabies vaccination and serologic monitoring practices are needed.

Metabolomics uncovers a link between inositol metabolism and osteosarcoma metastasis.

Cancer development and progression are characterized by complex molecular events. The acquisition of these events is primarily believed to result from alterations in gene and protein expression/function. Recent studies have also suggested the role of metabolic alterations, or "metabolic reprogramming," that may similarly contribute to these events. Indeed, our previous investigations in osteosarcoma (OS) identified metabolic changes uniquely linked to metastasis. Based on those findings, here we sought to build a more detailed understanding of the specific alterations in metabolites or metabolic pathways that may be responsible for the observed metastasis-associated metabolic alterations, suggested by gene expression data. This was pursued using a combination of high-throughput liquid- and gas-chromatography-based mass spectrometry (LC/MS and GC/MS) for a global metabolic profiling/subtraction of four pairs of high/low metastatic OS cell lines. By comparing the identity and level of the metabolites between high/low metastatic cells, several metabolic pathways were identified to be differentially activated, such as arginine, glutathione, inositol and fatty acid metabolic pathways. To further interrogate these results, we investigated the effects of inositol pathway dysregulation, through the exposure of metastatic OS cells to IP6 (inositol hexaphosphate). Although IP6 exposures had modest to minimal effects on cell proliferation, we observed reduced cellular glycolysis, down-regulation of PI3K/Akt signaling and suppression of OS metastatic progression. Collectively these data supported further investigation of metabolic sensitivities as anti-metastatic strategies in a clinical setting as well as investigation of altered metabolomics associated with metastatic progression.