Tigilanol tiglate is an oncolytic small molecule that induces immunogenic cell death and enhances the response of both target and non-injected tumors to immune checkpoint blockade.

BACKGROUND: Tigilanol tiglate (TT) is a protein kinase C (PKC)/C1 domain activator currently being developed as an intralesional agent for the treatment of various (sub)cutaneous malignancies. Previous work has shown that intratumoral (I.T.) injection of TT causes vascular disruption with concomitant tumor ablation in several preclinical models of cancer, in addition to various (sub)cutaneous tumors presenting in the veterinary clinic. TT has completed Phase I dose escalation trials, with some patients showing signs of abscopal effects. However, the exact molecular details underpinning its mechanism of action (MoA), together with its immunotherapeutic potential in oncology remain unclear. METHODS: A combination of microscopy, luciferase assays, immunofluorescence, immunoblotting, subcellular fractionation, intracellular ATP assays, phagocytosis assays and mixed lymphocyte reactions were used to probe the MoA of TT in vitro. In vivo studies with TT used MM649 xenograft, CT-26 and immune checkpoint inhibitor refractory B16-F10-OVA tumor bearing mice, the latter with or without anti-programmed cell death 1 (PD-1)/anti-cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) mAb treatment. The effect of TT at injected and non-injected tumors was also assessed. RESULTS: Here, we show that TT induces the death of endothelial and cancer cells at therapeutically relevant concentrations via a caspase/gasdermin E-dependent pyroptopic pathway. At therapeutic doses, our data demonstrate that TT acts as a lipotoxin, binding to and promoting mitochondrial/endoplasmic reticulum (ER) dysfunction (leading to unfolded protein responsemt/ER upregulation) with subsequent ATP depletion, organelle swelling, caspase activation, gasdermin E cleavage and induction of terminal necrosis. Consistent with binding to ER membranes, we found that TT treatment promoted activation of the integrated stress response together with the release/externalization of damage-associated molecular patterns (HMGB1, ATP, calreticulin) from cancer cells in vitro and in vivo, characteristics indicative of immunogenic cell death (ICD). Confirmation of ICD in vivo was obtained through vaccination and rechallenge experiments using CT-26 colon carcinoma tumor bearing mice. Furthermore, TT also reduced tumor volume, induced immune cell infiltration, as well as improved survival in B16-F10-OVA tumor bearing mice when combined with immune checkpoint blockade. CONCLUSIONS: These data demonstrate that TT is an oncolytic small molecule with multiple targets and confirms that cell death induced by this compound has the potential to augment antitumor responses to immunotherapy.

Use of angled-tip aspiration catheters is associated with a lower cost of thrombectomy in patients with acute ischemic stroke secondary to large and medium vessel occlusions.

BACKGROUND: Recently, the angled-tip Zoom™ aspiration catheters were introduced. The tip is designed to improve suction force for clot retrieval. We evaluated the possibility of reducing procedure costs when using angled-tip catheters and compared the safety and angiographic effectiveness of angled-tip versus straight-tip catheters. METHODS: We conducted a retrospective single-center cohort study involving patients with acute ischemic stroke due to large and medium vessel occlusions. The patients were divided into two groups: the post-Zoom group, in which angled-tip aspiration catheters were used and the pre-Zoom group, in which traditional straight-tip catheters were employed. RESULTS: A total of 163 patients were included; 95 (58.3%) in the pre-Zoom group and 68 (41.7%) in the post-Zoom group. The groups were well-matched at entry. The post-Zoom group demonstrated a significant decrease in mean procedure cost ($9728 vs. $12,127; p = 0.002), shorter time to achieve modified thrombolysis in cerebral infarction ≥2b reperfusion (38.30 min vs. 53.26 min; p = 0.018), and shorter puncture to procedure completion time (46.42 min vs. 62.38 min; p = 0.022). Additionally, the mean procedural cost when using the ADAPT technique supported by the Zoom catheters was significantly lower than the Solumbra technique ($5754 ± $2806 vs. $13,498 ± $3244, p < 0.001). There were no differences in the rate of hemorrhage between the pre-Zoom group (17.9%) and the post-Zoom group (20.6%), p = 0.690. CONCLUSION: The study demonstrated significant benefits, including cost reduction and shorter time to achieve reperfusion in patients treated with Zoom aspiration catheters. These findings support the use of angled-tip catheters in acute ischemic stroke management.

One bite or two? A comparison of single bite and double bite biopsy techniques in gastrointestinal endoscopy (BITES).

BACKGROUND AND AIM: Endoscopic biopsies can be taken using either single or double bite technique. In the single bite method, intubation time may be proportionately prolonged depending upon the number of biopsies taken. In contrast to this, double bite, though a greater number of biopsies may be taken per unit time, it may influence the quality of the biopsy specimen. The aim of the study was to compare these methods and to see if taking double bite has significant effect on the histological quality of the endoscopic biopsies. METHODS: A prospective, randomised and partly blind study (n = 135, M: 54%, age 21-91 years) divided into two equal arms to compare 144 procedures was conducted. Specimen were compared for time taken to size, depth, crush artefacts, necrosis, fragmentation, distortion, and epithelial stripping. Time taken to collect specimens was also recorded in the upper GI procedures. RESULTS: No significant difference was observed in the histological quality of single and double bite specimens (p < 0.05). However, DB took significantly less time (M = 88.5, SD ± 28.5) as compared to SB (M = 180, SD ± 55.9) (p < 0.05). CONCLUSIONS: There is no difference between the histological quality of DB and SB and the former technique takes less time, hence reducing intubation time.

Videoconference based training on diabetes technology for school nurses and staff: Pilot study.

PURPOSE: Children with diabetes spend a significant portion of time at school and in school-related activities and rely on school nurses for diabetes management support. Diabetes technologies are rapidly evolving, and there are no standardized competencies or training programs for school personnel providing diabetes care. DESIGN AND METHODS: A virtual diabetes education program was provided to school nurses and staff in 3 Florida school districts. Program feasibility was measured by attendance; acceptability was measured with a usability survey; and efficacy was measured by participants' improvements in scores on pre- and post-training knowledge assessments. Descriptive statistics were generated and improvements in knowledge were evaluated via t-test. P-values <0.05 were considered significant. RESULTS: Pilot survey data (n = 91) revealed high demand for diabetes technology and basic management education among school nurses and staff. Eighty-eight school personnel from 64 schools attended the training, with 67 participants completing the demographic survey and at least one of the pre- and post-training assessments. Post-test scores demonstrated mean + 10.6% absolute improvement on the diabetes technology subscale, +11.5% on the basic management subscale, and + 10.9% on the ketone management subscale, all p < 0.001. Fifty-three participants completed the usability survey with 92% reporting they benefitted from training. CONCLUSIONS: Virtual training is feasible and acceptable for delivering diabetes technology education to large numbers of school personnel. Study results demonstrate improved diabetes knowledge. PRACTICE IMPLICATIONS: Establishing a standardized training program on diabetes technology for school personnel can optimize diabetes care in the school setting.

Effects of Leg Length, Sex, Laterality, and the Intermediate Femoral Cutaneous Nerve on Infrapatellar Innervation.

Background: An iatrogenic injury to the infrapatellar branch of the saphenous nerve (IPBSN) is a common precipitant of postoperative knee pain and hypoesthesia. Purpose: To locate potential safe zones for incision by observing the patterns and pathway of the IPBSN while examining the relationship of its location to sex, laterality, and leg length. Study Design: Descriptive laboratory study. Methods: A total of 107 extended knees from 55 formalin-embalmed cadaveric specimens were dissected. The nerve was measured from palpable landmarks: the patella at the medial (point A) and lateral (point B) borders of the patellar ligament, the medial border of the patellar ligament at the patellar apex (point C) and tibial plateau (point D), the medial epicondyle (point E), and the anterior border of the medial collateral ligament at the tibial plateau (point F). The safe zone was defined as 2 SDs from the mean. Results: Findings indicated significant correlations between leg length and height (r P = 0.832; P < .001) as well as between leg length and vertical measurements (≥45°) from points A and B to the IPBSN (r P range, 0.193-0.285; P range, .004-.049). Male specimens had a more inferior maximum distance from point A to the intersection of the IPBSN and the medial border of the patellar ligament compared with female specimens (6.17 vs 5.28 cm, respectively; P = .049). Right knees had a more posterior IPBSN from point F compared with left knees (-0.98 vs-0.02 cm, respectively; P = .048). The majority of knees (62.6%; n = 67) had a nerve emerging that penetrated the sartorius muscle. Additionally, 32.7% (n = 35) had redundant innervation, and 25.2% (n = 27) had contribution from the intermediate femoral cutaneous nerve (IFCN). Conclusion: We identified no safe zone. Significant innervation redundancy with a substantial contribution to the infrapatellar area from the IFCN was noted and contributed to the expansion of the danger zone. Clinical Relevance: The location of incision and placement of arthroscopic ports might not be as crucial in postoperative pain management as an appreciation of the variance in infrapatellar innervation. The IFCN is a common contributor. Its damage could explain pain refractory to SN blocks and therefore influence anesthetic and analgesic decisions.

Provision and standards of care for treatment and follow-up of patients with Autoimmune Hepatitis (AIH).

Background: Autoimmune hepatitis (AIH) is a substantial UK health burden, but there is variation in care, facilities and in opinion regarding management. We conducted an audit of service provision and care of patients with AIH in 28 UK hospitals. Methods: Centres provided information about staffing, infrastructure and patient management (measured against predefined guideline-based standards) via a web-based data collection tool. Results: Hospitals (14 university hospitals (UHs), 14 district general hospitals (DGHs)) had median (range) of 8 (3-23) gastroenterologists; including 3 (0-10) hepatologists. Eight hospitals (29%, all DGHs) had no hepatologist. In individual hospital departments, there were 50% (18-100) of all consultants managing AIH: in DGH's 92% (20-100) vs 46% (17-100) in UHs. Specialist nurses managed AIH in only 18%. Seventeen (61%) hospitals had a histopathologist with a liver interest, these were more likely to find rosettes than those without (172/795 vs 50/368; p<0.001).Of 999 steroid-treated patients with ≥12 months follow-up, 25% received steroids for <12 months. After 1 year of treatment, 82% of patients achieved normal serum alanine aminotransaminase (ALT); this was higher in UHs than DGHs. Three-monthly liver blood tests were inadequately recorded in 26%. Of potentially eligible patients with liver decompensation, transplantation was apparently not considered in 5% (n=7). The same standards were attained in different types of hospital. Conclusion: Management of AIH in UK hospitals is often shared between most gastroenterologists. Blood test monitoring and treatment duration are not always in line with recommendations. Some eligible patients with decompensation are not discussed with transplant teams. Care might be improved by expanding specialist input and management by fewer designated consultants.

Treatment and Outcome of Autoimmune Hepatitis (AIH): Audit of 28 UK centres.

BACKGROUND: With few data regarding treatment and outcome of patients with AIH outside of large centres we present such a study of patients with AIH in 28 UK hospitals of varying size and facilities. METHODS: Patients with AIH were identified in 14 University and 14 District General hospitals; incident cases during 2007-2015 and prevalent cases, presenting 2000-2015. Treatment and outcomes were analysed. RESULTS: In 1267 patients with AIH, followed-up for 3.8(0-15) years, 5- and 10-year death/transplant rates were 7.1+0.8% and 10.1+1.3% (all-cause) and 4.0+0.6% and 5.9+1% (liver-related) respectively. Baseline parameters independently associated with death/transplantation for all-causes were: older age, vascular/respiratory co-morbidity, cirrhosis, decompensation, platelet count, attending transplant centre and for liver-related: the last four of these and peak bilirubin All-cause and liver-related death/transplantation was independently associated with: non-treatment with corticosteroids, non-treatment with a steroid-sparing agent (SSA), non-treatment of asymptomatic or non-cirrhotic patients and initial dose of Prednisolone >35mg/0.5mg/kg/day (all-cause only), but not with type of steroid (Prednisolone versus Budesonide) or steroid duration beyond 12-months. Subsequent all-cause and liver-death/transplant rates showed independent associations with smaller percentage fall in serum ALT after 1 and 3-months, but not with failure to normalise levels over 12-months. CONCLUSIONS: We observed higher death/transplant rates in patients with AIH who were untreated with steroids (including asymptomatic or non-cirrhotic sub-groups), those receiving higher Prednisolone doses and those who did not receive an SSA. Similar death/transplant rates were seen in those receiving Prednisolone or Budesonide, those continuing steroids after 12-months and patients attaining normal ALT within 12-months versus not.

Mapping a Danger Zone of the Dorsal Nerve of the Clitoris: Implications in Female Cosmetic Genital Surgery.

BACKGROUND: The literature regarding the route of the dorsal nerve of the clitoris is sparse and lacks surgical focus. With an increasing number of procedures being performed on the labia, it is important to elucidate the route and note any variation from normal of the nerve. METHODS: Fifty-one cadavers were dissected to yield 97 dorsal nerve of the clitoris samples. Measurements were taken from (1) the dorsal nerve of the clitoris penetration point of the perineal membrane to the urethra, (2) the nerve's penetration point of the perineal membrane to the pubic bone, (3) the angle of the clitoris to the branch point of the dorsal nerve of the clitoris, and (4) the branch point of the nerve to the distalmost point of the glans clitoris. Any anomalous branching patterns of the dorsal nerve of the clitoris were recorded and classified. RESULTS: The means and standard deviations of each measurement were used to create a surgical danger zone. The mean of each measurement was (1) 34.63 mm, (2) 5.74 mm, (3) -3.07 mm, and (4) 30.40 mm, respectively. In addition, six distinct branching patterns were observed, organized, and classified based on the location and number of branches observed. CONCLUSIONS: The dorsal nerve of the clitoris has multiple branching patterns and typically travels along the same course in most women. Further investigation of the course and three-dimensional position of the dorsal nerve of the clitoris is warranted to preserve sexual sensation as the frequency of procedures involving the female pudendum increases.

Activation of PKC supports the anticancer activity of tigilanol tiglate and related epoxytiglianes.

The long-standing perception of Protein Kinase C (PKC) as a family of oncoproteins has increasingly been challenged by evidence that some PKC isoforms may act as tumor suppressors. To explore the hypothesis that activation, rather than inhibition, of these isoforms is critical for anticancer activity, we isolated and characterized a family of 16 novel phorboids closely-related to tigilanol tiglate (EBC-46), a PKC-activating epoxytigliane showing promising clinical safety and efficacy for intratumoral treatment of cancers. While alkyl branching features of the C12-ester influenced potency, the 6,7-epoxide structural motif and position was critical to PKC activation in vitro. A subset of the 6,7-epoxytiglianes were efficacious against established tumors in mice; which generally correlated with in vitro activation of PKC. Importantly, epoxytiglianes without evidence of PKC activation showed limited antitumor efficacy. Taken together, these findings provide a strong rationale to reassess the role of PKC isoforms in cancer, and suggest in some situations their activation can be a promising strategy for anticancer drug discovery.

Wound formation, wound size, and progression of wound healing after intratumoral treatment of mast cell tumors in dogs with tigilanol tiglate.

BACKGROUND: Tigilanol tiglate (TT) is a novel small molecule for intratumoral treatment of nonmetastatic mast cell tumors (MCTs) in dogs. In a randomized controlled clinical study, 75% of dogs that received a single TT treatment achieved complete resolution of the MCT by 28 days, with no recurrence in 93% of dogs at 84 days. Critical to TT's efficacy was the area of the wound (tissue deficit) after slough of the necrotic tumor relative to pretreatment tumor volume. OBJECTIVES: To analyze data collected during the previous study to (a) describe wounds after slough of treated MCTs and (b) identify determinants of wound area and speed of wound healing. METHODS: Wound presence, condition, and area were determined from clinical records of 117 dogs over 84 days after a single intratumoral TT treatment. RESULTS: Tumor slough occurred 3 to 14 days after treatment, exposing granulation tissue in the wound bed. Wound area after tumor slough in general was related to pretreatment tumor volume, with maximal recorded wound area fully evident in 89% of dogs by day 7. In dogs achieving complete tumor resolution, all wounds were left to heal by secondary intention. Bandaging and other wound management interventions only were required in 5 dogs. Time to healing (ie, full re-epithelialization of treatment site) depended on wound area and location on the body, with most wounds being fully healed between 28 and 42 days after treatment. CONCLUSIONS: Wound area and healing after slough of TT-treated tumors follow a consistent clinical pattern for most dogs.

Key Anatomical Clarifications for the Marginal Mandibular Branch of the Facial Nerve: Clinical Significance for the Plastic Surgeon.

BACKGROUND: The marginal mandibular branch (MMBr) of the facial nerve is the least likely to recover from injury due to infrequent anastomosis with other branches. The MMBr has been described as coursing superior to the inferior border of the mandible. However, studies have reported variations in its location in embalmed and fresh specimens. It has been postulated that the embalming process may effect its anatomic position. OBJECTIVES: The aim of this study was to re-evalulate the location of the MMBr relative to the inferior border of the mandible in both fresh and embalmed cadavers, and investigate variation in its position with sex, side of the face, and age. METHODS: Superficial fascial planes were dissected to reveal the MMBr and its anatomic relations. Distance between the most inferior branch of the MMBr and the antegonial notch were measured bilaterally. The most inferior position of the MMBr between the antegonial notch and gonion was measured. Fresh heads were used as a comparison, with an additional measurement taken of the distance between the MMBr and the gonial angle. RESULTS: The MMBr was located inferior to the border of the mandible (90.3%) more often than above (9.6%). No significant differences were found between fresh and embalmed cadavers, sex, side of body, or age (P > 0.05). No significant difference was found between intact cadavers and fresh heads (P > 0.05). CONCLUSIONS: This study confirms and describes reliable landmarks for safety zones for the MMBr during plastic and reconstructive surgery of the lower face and upper neck. These data add reliability to studies that have investigated nerve locations in embalmed cadavers.

Randomized controlled clinical study evaluating the efficacy and safety of intratumoral treatment of canine mast cell tumors with tigilanol tiglate (EBC-46).

OBJECTIVE: To evaluate the efficacy and safety of tigilanol tiglate (TT) for local intratumoral treatment of mast cell tumors (MCTs) in dogs. METHODS: A randomized controlled clinical study in 2 phases involving 123 dogs with cytologically diagnosed MCT. Phase 1 compared 81 TT-treated dogs with 42 control dogs; phase 2 allowed TT treatment of control dogs and retreatment of dogs that failed to achieve tumor resolution after TT treatment in phase 1. Tigilanol tiglate (1 mg/mL) was injected intratumorally with dose based on tumor volume. Concomitant medications were used to minimize potential for MCT degranulation. Modified response evaluation criteria in solid tumors were used to evaluate treatment response at 28 and 84 days. Adverse events and quality of life were also assessed. RESULTS: A single TT treatment resulted in 75% complete response (CR) (95% confidence interval [CI] = 61-86) by 28 days, with no recurrence in 93% (95% CI = 82-97) of dogs by 84 days. Eight TT-treated dogs that did not achieve CR in phase 1 achieved CR after retreatment, increasing the overall CR to 88% (95% CI = 77-93). Control dogs had 5% CR (95% CI = 1-17) at 28 days. Wound formation after tumor slough and wound size relative to tumor volume were strongly associated with efficacy. Adverse events typically were low grade, transient, and directly associated with TT's mode of action. CONCLUSIONS: Tigilanol tiglate is efficacious and well tolerated, providing a new option for the local treatment of MCTs in dogs.

Etiology and Comorbidity Diagnoses Effect on Outcomes for Patients Undergoing Endoscopic Retrograde Cholangiopancreatography.

Introduction Endoscopic retrograde cholangiopancreatography (ERCP) is now the first-line approach to treating choledocholithiasis. As a minimally invasive procedure, it is considered relatively safe but still entails a higher risk than other routine endoscopic procedures. This study aims to look at possible patient etiologies and comorbidities that may affect patient outcomes.  Methods This study used the Nationwide Inpatient Sample (NIS) from the years 2012 - 2015 to collect anonymous patient data through the use of International Classification of Diseases, Ninth Revision (ICD-9) codes. Specific codes were used to determine the top five etiologies (or presenting diagnosis) for patients who had this surgery and to separate outpatients with specific comorbidity diagnoses. The IBM Statistical Package for Social Sciences (SPSS) (IBM SPSS Statistics, Armonk, NY) was then used to compare patients with these diagnoses or etiologies to those without to measure differences in patient outcomes, such as mortality, length of stay, and total charges. Results Patients who had an etiological diagnosis of acute kidney failure had worse outcomes than patients who were admitted for ERCP without that etiological diagnosis. There were also specific comorbidity diagnoses that were noted to have worse patient outcomes, including congestive heart failure, diabetes mellitus with complications, a coagulopathy disorder, anemia, or chronic liver disease. Additionally, patients who had both acute kidney disease and chronic liver disease had the worst outcomes. Conclusions This study highlights the need to understand all patient risk factors before having them undergo ERCP, especially in the setting of scheduled surgery. Working to control these factors before surgery can increase the possibility of avoiding negative outcomes like mortality, increased patient costs, and increased length of stay.

EBC-232 and 323: A Structural Conundrum Necessitating Unification of Five In Silico Prediction and Elucidation Methods.

Structurally unique halimanes EBC-232 and EBC-323, isolated from the Australian rainforest plant Croton insularis, proved considerably difficult to elucidate. The two diastereomers, which consist an unusual oxo-6,7-spiro ring system fused to a dihydrofuran, were solved by unification and consultation of five in silico NMR elucidation and prediction methods [i.e., ACDLabs, olefin strain energy (OSE), DP4, DU8+ and TD DFT CD]. Structure elucidation challenges of this nature are prime test case examples for empowering future AI learning in structure elucidation.

Anatomic Variations From 120 Mental Nerve Dissections: Lessons for Transoral Thyroidectomy.

BACKGROUND: Transoral endoscopic thyroidectomy vestibular approach (TOETVA) is a promising technique for eliminating a neck incision. A new risk of TOETVA is the potential for injury to the mental nerves during placement of three oral endoscopic ports. A better understanding of the variations in mental nerve anatomy is needed to inform safer TOETVA technique. MATERIALS AND METHODS: We performed 120 dissections of mental nerve branches exiting the mental foramen in 60 human cadavers. Anatomic distances and relationships of the foramen to the midline were evaluated. Mental nerve branching patterns were studied and compared with previously reported classification systems to determine surgical safe zones free of nerve branches. RESULTS: The mean midline-to-mental foramen distance was 29.2 ± 3.3 mm, with high variability across individuals (18.8-36.8 mm). There were differences in this distance between the left and right foramina (29.8 ± 3.2 versus 28.8 ± 3.3 mm, P = 0.03). All mental nerve branches exiting the mental foramen distributed medially. The branching patterns were classified into eight distinct categories, three of which are previously undescribed. One of these novel patterns, occurring in 9.2% of cases, had a dense and wide clustering of branches traveling toward the midline. CONCLUSIONS: The location of the mental foramen and mental nerve branching patterns demonstrate high variability. To avoid mental nerve injury in TOETVA, we identify a safe zone for lateral port placement lateral to the plane of the mental foramen. Placement and extension of the middle port incision should proceed with caution, as clustering of mental nerve branches in this area can frequently be present.

Novel epoxy-tiglianes stimulate skin keratinocyte wound healing responses and re-epithelialization via protein kinase C activation.

Epoxy-tiglianes are a novel class of diterpene esters. The prototype epoxy-tigliane, EBC-46 (tigilanol tiglate), possesses potent anti-cancer properties and is currently in clinical development as a local treatment for human and veterinary cutaneous tumors. EBC-46 rapidly destroys treated tumors and consistently promotes wound re-epithelialization at sites of tumor destruction. However, the mechanisms underlying these keratinocyte wound healing responses are not completely understood. Here, we investigated the effects of EBC-46 and an analogue (EBC-211) at 1.51 nM-151 µM concentrations, on wound healing responses in immortalized human skin keratinocytes (HaCaTs). Both EBC-46 and EBC-211 (1.51 nM-15.1 µM) accelerated G0/G1-S and S-G2/M cell cycle transitions and HaCaT proliferation. EBC-46 (1.51-151 nM) and EBC-211 (1.51 nM-15.1 µM) further induced significant HaCaT migration and scratch wound repopulation. Stimulated migration/wound repopulation responses were even induced by EBC-46 (1.51 nM) and EBC-211 (1.51-151 nM) with proliferation inhibitor, mitomycin C (1 µM), suggesting that epoxy-tiglianes can promote migration and wound repopulation independently of proliferation. Expression profiling analyses showed that epoxy-tiglianes modulated keratin, DNA synthesis/replication, cell cycle/proliferation, motility/migration, differentiation, matrix metalloproteinase (MMP) and cytokine/chemokine gene expression, to facilitate enhanced responses. Although epoxy-tiglianes down-regulated established cytokine and chemokine agonists of keratinocyte proliferation and migration, enhanced HaCaT responses were demonstrated to be mediated via protein kinase C (PKC) phosphorylation and significantly abrogated by pan-PKC inhibitor, bisindolylmaleimide-1 (BIM-1, 1 µM). By identifying how epoxy-tiglianes stimulate keratinocyte healing responses and re-epithelialization in treated skin, our findings support the further development of this class of small molecules as potential therapeutics for other clinical situations associated with impaired re-epithelialization, such as non-healing skin wounds.

Dose Characterization of the Investigational Anticancer Drug Tigilanol Tiglate (EBC-46) in the Local Treatment of Canine Mast Cell Tumors.

Mast cell tumor (MCT) is the most common cutaneous neoplasm in dogs and wide surgical resection is the current first-line treatment. However, recurrence is common and often requires more specialist and expensive therapies. Tigilanol tiglate is a novel small molecule drug delivered by intratumoral injection that is currently under development to provide a new option for treating MCT. The aim of this study was to characterize a safe and effective dose of tigilanol tiglate for canine MCT and to gather preliminary data on the drug's pharmacokinetics. A multicenter, open-label, uncontrolled, non-randomized, dose de-escalation design was used. Eligibility was MCT stage I/IIa and a tumor size of 0.1-6.0 cm3. Dosing was based on tumor size (50% v/v tumor) and 3 drug concentrations (1.0, 0.5, 0.2 mg/mL) were evaluated. Twenty-seven dogs were treated in 3 dose de-escalation cohorts (10, 10, and 7 dogs, respectively). Efficacy at 21 days was defined using international accepted solid tumor response criteria (RECIST). Greatest efficacy (90% complete response) was observed at the highest drug concentration (1.0 mg/mL) and adverse events were generally low grade, mild and transient, and directly associated with the mode of action of the drug. Hematological and serum biochemistry were generally unremarkable with plasma concentration curves typical of a non-intravenous parenteral medication. Intratumoral treatment of MCT with tigilanol tiglate at a concentration of 1.0 mg/mL was highly efficacious and well-tolerated. These results support the drug's further development for the treatment of MCT and other solid tumors.

New Casbanes and the First trans-Cyclopropane seco-Casbane from the Australian Rainforest Plant Croton insularis.

Investigation of the Australian rainforest plant Croton insularis revealed seven new cis-, two new trans-cyclopropane casbanes, and the first trans-cyclopropane seco-casbane. The relative configuration of the cyclopropane moiety for all compounds (EBC-182, 217, 218, 220, 343, 357, 358, 361, 365, 373; EBC=EcoBiotics Compound) was assigned using 13 C NMR data. Comparison of the experimental electronic circular dichroism (ECD) spectra with the theoretical curves, calculated by TD-DFT at the B3LYP/6-31+G**//B3LYP/6-31+G** level, in conjunction with NOE data afforded the absolute configuration. EBC-180, 181 and 220 displayed potent activity against cervical carcinoma (HeLa cells).

Diagnosis, presentation and initial severity of Autoimmune Hepatitis (AIH) in patients attending 28 hospitals in the UK.

BACKGROUND & AIMS: There is limited information regarding patients with AIH outside relatively few large centres. We describe here the presenting features of patients with AIH, collected as part of an audit involving 28 UK hospitals. METHODS: Patients (incident since 1/1/2007 or prevalent since 1/1/2000) were ≥18 years and either met 1999 International AIH Group (IAIHG) diagnostic criteria (n = 1164), or received immunosuppressive therapy for clinically diagnosed AIH (n = 103). RESULTS: Of 1267 patients (80% women, 91% Caucasian, age (median(range)) 55(8-86) years, 0.5% had acute viral hepatitis (CMV/EBV/HEV); 2% were taking Nitrofurantoin and 0.7% Khat. Twenty-one percent had clinical decompensation and/or a MELD score of >15. Time from first abnormal liver tests to diagnosis was ≥1 year in 19% and was longer in jaundiced vs non-jaundiced patients. HBV and HCV serology were undocumented in 4%, serum immunoglobulins in 31% and autoantibodies in 11%-27%. When documented, ≥1 antibody was present in 83%. LKM-1-positive and autoantibody-negative patients had more severe disease. Histological cirrhosis was reported in 23%, interface hepatitis 88%, predominant lymphocytes/plasma cells 75%, rosettes 19% and emperipolesis 0.4%. Only 65% of those meeting 1999 IAIHG criteria also met simplified IAIHG criteria. University Hospitals compared to District General Hospitals, were more likely to report histological features of AIH. CONCLUSIONS: This cohort from across the UK is older than other multicentre AIH cohorts. One-fifth had decompensation or MELD >15. Diagnosis was delayed in 19%, diagnostic testing was incomplete in one-third and rosettes and emperipolesis were infrequently reported.

The First Casbane Hydroperoxides EBC-304 and EBC-320 from the Australian Rainforest.

Investigation of the Australian rainforest plant Croton insularis led to isolation of the first casbane hydroperoxide diterpenes EBC-304 and EBC-320. Extensive DFT and electronic circular dichroism (ECD) calculations in combination with 2D NMR spectroscopy determined the absolute configurations. EBC-304 and EBC-320 both display significant cytotoxicity.