GHSR Deletion in ß-Cells of Male Mice: Ineffective in Obesity, but Effective in Protecting against Streptozotocin-Induced ß-Cell Injury in Aging.

Insulin secretion from pancreatic ß cells is a key pillar of glucose homeostasis, which is impaired under obesity and aging. Growth hormone secretagogue receptor (GHSR) is the receptor of nutrient-sensing hormone ghrelin. Previously, we showed that ß-cell GHSR regulated glucose-stimulated insulin secretion (GSIS) in young mice. In the current study, we further investigated the effects of GHSR on insulin secretion in male mice under diet-induced obesity (DIO) and streptozotocin (STZ)-induced ß-cell injury in aging. ß-cell-specific-Ghsr-deficient (Ghsr-ßKO) mice exhibited no glycemic phenotype under DIO but showed significantly improved ex vivo GSIS in aging. We also detected reduced insulin sensitivity and impaired insulin secretion during aging both in vivo and ex vivo. Accordingly, there were age-related alterations in expression of glucose transporter, insulin signaling pathway, and inflammatory genes. To further determine whether GHSR deficiency affected ß-cell susceptibility to acute injury, young, middle-aged, and old Ghsr-ßKO mice were subjected to STZ. We found that middle-aged and old Ghsr-ßKO mice were protected from STZ-induced hyperglycemia and impaired insulin secretion, correlated with increased expression of insulin signaling regulators but decreased pro-inflammatory cytokines in pancreatic islets. Collectively, our findings indicate that ß-cell GHSR has a major impact on insulin secretion in aging but not obesity, and GHSR deficiency protects against STZ-induced ß-cell injury in aging.

Measuring intramolecular connectivity in long RNA molecules using two-dimensional DNA patch-probe arrays.

We describe a simple method to infer intramolecular connections in a population of long RNA molecules in vitro. First we add DNA oligonucleotide "patches" that perturb the RNA connections, then we use a microarray containing a complete set of DNA oligonucleotide "probes" to record where perturbations occur. The pattern of perturbations reveals couplings between different regions of the RNA sequence, from which we infer connections as well as their prevalences in the population. We validate this patch-probe method using the 1,058-nucleotide RNA genome of satellite tobacco mosaic virus (STMV), which has previously been shown to have multiple long-range connections. Our results not only indicate long duplexes that agree with previous structures but also reveal the prevalence of competing connections. Together, these results suggest that globally-folded and locally-folded structures coexist in solution. We show that the prevalence of connections changes when pseudouridine, an important component of natural and synthetic RNA molecules, is substituted for uridine in STMV RNA.

Sodium-Glucose Cotransporter 2 Inhibitors in Frail Patients with Heart Failure: Clinical Experience of a Heart Failure Unit.

OBJECTIVE: The objective of this study was to determine the difference in tolerance for sodium glucose cotransporter 2 inhibitors between patients with heart failure classified as frail according to the FRAIL questionnaire, compared to those with heart failure without frailty. METHODS: A prospective cohort study was performed between 2021 and 2022 that included patients with heart failure at a heart failure unit in Bogotá who were being treated with a sodium glucose cotransporter 2 inhibitor. Clinical and laboratory data were collected during an initial visit and 12-48 weeks after that. The FRAIL questionnaire was applied to all participants through a phone call or during the follow-up visit. The primary outcome was the adverse effect rate and as a secondary outcome we compared the estimated glomerular filtration rate change between frail and non-frail patients. RESULTS: One hundred and twelve patients were included in the final analysis. Frail patients had a more than twice increased risk of having adverse effects (95% confidence interval 1.5-3.9). Age was also a risk factor for the appearance of these.  The estimated glomerular filtration rate decrease was inversely correlated with the age, left ventricular ejection fraction, and renal function before the use of sodium glucose cotransporter 2 inhibitors. CONCLUSIONS: When prescribing in heart failure, it is important to remember that frail patients are more likely to have adverse effects with the use of sodium glucose cotransporter 2 inhibitors, of which the most common are those related to osmotic diuresis. Nonetheless, these do not appear to increase the risk of discontinuation or abandonment of therapy in this population.

Video Validation of Tri-Axial Accelerometer for Monitoring Zoo-Housed Tamandua tetradactyla Activity Patterns in Response to Changes in Husbandry Conditions.

Accelerometers are a technology that is increasingly used in the evaluation of animal behaviour. A tri-axial accelerometer attached to a vest was used on Tamandua tetradactyla individuals (n = 10) at Biodiversity Park. First, the influence of using a vest on the animals' behaviour was evaluated (ABA-type: A1 and A2, without a vest; B, with a vest; each stage lasted 24 h), and no changes were detected. Second, their behaviour was monitored using videos and the accelerometer simultaneously (experimental room, 20 min). The observed behaviours were correlated with the accelerometer data, and summary measures (X, Y and Z axes) were obtained. Additionally, the overall dynamic body acceleration was calculated, determining a threshold to discriminate activity/inactivity events (variance = 0.0055). Then, based on a 24 h complementary test (video sampling every 5 min), the sensitivity (85.91%) and precision (100%) of the accelerometer were assessed. Animals were exposed to an ABA-type experimental design: A1 and A2: complex enclosure; B: decreased complexity (each stage lasted 24 h). An increase in total activity (%) was revealed using the accelerometer (26.15 ± 1.50, 29.29 ± 2.25, and 35.36 ± 3.15, respectively). Similar activity levels were detected using video analysis. The results demonstrate that the use of the accelerometer is reliable to determine the activity. Considering that the zoo-housed lesser anteaters exhibit a cathemeral activity pattern, this study contributes to easily monitoring their activities and responses to different management procedures supporting welfare programs, as well as ex situ conservation.

Growth of Nitrogen Incorporated Ultrananocrystalline Diamond Coating on Graphite by Hot Filament Chemical Vapor Deposition.

This article shows the results of experiments to grow Nitrogen incorporated ultrananocrystalline diamond (N-UNCD) films on commercial natural graphite (NG)/Cu anodes by hot chemical vapor deposition (HFCVD) using a gas mixture of Ar/CH4/N2/H2. The experiments focused on studying the effect of the pressure in the HFCVD chamber, filament-substrate distance, and temperature of the substrate. It was found that a substrate distance of 3.0 cm and a substrate temperature of 575 C were optimal to grow N-UNCD film on the graphite surface as determined by Raman spectroscopy, SEM, and TEM imaging. XPS analysis shows N incorporation through the film. Subsequently, the substrate surface temperature was increased using a heater, while keeping the substrate-filament distance constant at 3.0 cm. In this case, Raman spectra and SEM images of the substrate surface showed a major composition of graphite in the film as the substrate-surface temperature increased. Finally, the process pressure was increased to 10 Torr where it was seen that the growth of N-UNCD film occurred at 2.0 cm at a substrate temperature of 675 C. These results suggest that as the process pressure increases a smaller substrate-filament distance and consequently a higher substrate surface temperature can still enable the N-UNCD film growth by HFCVD. This effect is explained by a mean free path analysis of the main precursors H2 and CH3 molecules traveling from the filament to the surface of the substrate The potential impact of the process developed to grow electrically conductive N-UNCD films using the relatively low-cost HFCVD process is that this process can be used to grow N-UNCD films on commercial NG/Cu anodes for Li-ion batteries (LIBs), to enable longer stable capacity energy vs. charge/discharge cycles.

Single-particle studies of the effects of RNA-protein interactions on the self-assembly of RNA virus particles.

Understanding the pathways by which simple RNA viruses self-assemble from their coat proteins and RNA is of practical and fundamental interest. Although RNA-protein interactions are thought to play a critical role in the assembly, our understanding of their effects is limited because the assembly process is difficult to observe directly. We address this problem by using interferometric scattering microscopy, a sensitive optical technique with high dynamic range, to follow the in vitro assembly kinetics of more than 500 individual particles of brome mosaic virus (BMV)-for which RNA-protein interactions can be controlled by varying the ionic strength of the buffer. We find that when RNA-protein interactions are weak, BMV assembles by a nucleation-and-growth pathway in which a small cluster of RNA-bound proteins must exceed a critical size before additional proteins can bind. As the strength of RNA-protein interactions increases, the nucleation time becomes shorter and more narrowly distributed, but the time to grow a capsid after nucleation is largely unaffected. These results suggest that the nucleation rate is controlled by RNA-protein interactions, while the growth process is driven less by RNA-protein interactions and more by protein-protein interactions and intraprotein forces. The nucleated pathway observed with the plant virus BMV is strikingly similar to that previously observed with bacteriophage MS2, a phylogenetically distinct virus with a different host kingdom. These results raise the possibility that nucleated assembly pathways might be common to other RNA viruses.

Computer tool with intelligent behavior for the optimal preliminary design in non-braced structural steel frame.

The optimization of civil structures is a technique whose purpose is to efficiently use the materials that make up the structural systems based on previously established restrictions and objectives. The use and development of these techniques has been closely linked to technological advance since, through the use of computer equipment, complex mathematical models can be solved with low cost and time. This article presents OPS Design v2.0, a computer tool that allows obtaining a preliminary optimal distribution of metallic structural profiles in a Non-Braced Frame System (OMF: Ordinary Moment Frame). The optimization model implemented in OPS Design v2.0 seeks to minimize the number of different profiles and the structure's own weight in order to reduce the construction complexity and the weight per linear meter (costs in quantities of material). To evaluate its effectiveness, a case study was developed where it was concluded that the designs produced by the application are more efficient than those obtained by commercial tools, thus reducing the computational expense and time used by designers in iterative processes that are carried out in the initial phases project.

Diverse and Complementary Effects of Ghrelin and Obestatin.

Ghrelin and obestatin are two "sibling proteins" encoded by the same preproghrelin gene but possess an array of diverse and complex functions. While there are ample literature documenting ghrelin's functions, the roles of obestatin are less clear and controversial. Ghrelin and obestatin have been perceived to be antagonistic initially; however, recent studies challenge this dogma. While they have opposing effects in some systems, they function synergistically in other systems, with many functions remaining debatable. In this review, we discuss their functional relationship under three "C" categories, namely complex, complementary, and contradictory. Their functions in food intake, weight regulation, hydration, gastrointestinal motility, inflammation, and insulin secretion are complex. Their functions in pancreatic beta cells, cardiovascular, muscle, neuroprotection, cancer, and digestive system are complementary. Their functions in white adipose tissue, thermogenesis, and sleep regulation are contradictory. Overall, this review accumulates the multifaceted functions of ghrelin and obestatin under both physiological and pathological conditions, with the intent of contributing to a better understanding of these two important gut hormones.

ß Cell GHS-R Regulates Insulin Secretion and Sensitivity.

Growth hormone secretagogue receptor (GHS-R) is widely known to regulate food intake and adiposity, but its role in glucose homeostasis is unclear. In this study, we investigated the expression of GHS-R in mouse pancreatic islets and its role in glycemic regulation. We used Ghsr-IRES-tauGFP mice, with Green Fluorescent Protein (GFP) as a surrogate for GHS-R, to demonstrate the GFP co-localization with insulin and glucagon expression in pancreatic islets, confirming GHS-R expression in ß and α cells. We then generated ß-cell-specific GHSR-deleted mice with MIP-Cre/ERT and validated that GHS-R suppression was restricted to the pancreatic islets. MIP-Cre/ERT;Ghsrf/f mice showed normal energy homeostasis with similar body weight, body composition, and indirect calorimetry profile. Interestingly, MIP-Cre/ERT;Ghsrf/f mice exhibited an impressive phenotype in glucose homeostasis. Compared to controls, MIP-Cre/ERT;Ghsrf/f mice showed lower fasting blood glucose and insulin; reduced first-phase insulin secretion during a glucose tolerance test (GTT) and glucose-stimulated insulin secretion (GSIS) test in vivo. The isolated pancreatic islets of MIP-Cre/ERT;Ghsrf/f mice also showed reduced insulin secretion during GSIS ex vivo. Further, MIP-Cre/ERT;Ghsrf/f mice exhibited improved insulin sensitivity during insulin tolerance tests (ITT). Overall, our results confirmed GHS-R expression in pancreatic ß and α cells; GHS-R cell-autonomously regulated GSIS and modulated systemic insulin sensitivity. In conclusion, ß cell GHS-R was an important regulator of glucose homeostasis, and GHS-R antagonists may have therapeutic potential for Type 2 Diabetes.

Non-Invasive Assessment of the Seasonal Stress Response to Veterinary Procedures and Transportation of Zoo-Housed Lesser Anteater (Tamandua tetradactyla).

Management procedures affect behavioural and physiological stress responses of wild mammals under human care. According to the Reactive Scope Model, normal values are presumed to exist within predictive and reactive ranges. First, stress parameters of zoo-housed adult Tamandua tetradactyla were evaluated in winter and summer (29 days each), determining the level of behaviour and/or physiological parameters needed to respond to predictable environmental changes. Secondly, the effects of veterinary procedures and transportation were studied in both seasons. Non-invasive methods were applied, assessing behaviour through videos and adrenocortical activity by faecal glucocorticoid metabolites (FGMs). Lesser anteaters exhibited seasonality (summer > winter) in some behavioural parameters, such as nocturnal activities, as well as in the activity cycle (e.g., acrophase) and FGMs. A veterinary check elicited an increase in total activity (TA), natural behaviours and repetitive locomotion and affected the activity cycle, particularly in summer. Transport produced changes in TA, nocturnal and natural activity and some variables of the activity cycle, mostly during summer. Although the effects of routine management procedures were different from each other and presumably stressful, they elicited changes only at the behavioural level, which was greater during summer. The differences observed according to non-invasive methodologies highlight the importance of a multidisciplinary approach in this context and suggest that it is unlikely that individual welfare was affected.

ACV y covid-19: una revisión de los estudios observacionales publicados en época de pandemia / Stroke and COVID-19: A review of observational studies published during the pandemic

RESUMEN INTRODUCCIÓN: En marzo 2020 la Organización Mundial de la Salud decretó la pandemia por covid-19. Se han informado casos de ACV relacionados con esta infección viral. OBJETIVOS: Conocer la experiencia de diferentes partes del mundo respecto al ACV y covid-19 con el fin de mejorar el reconocimiento y saber qué hacer cuando se empiecen a presentar estos pacientes en nuestro medio. MÉTODOS: Se hizo una revisión de los estudios observacionales disponibles utilizando PubMed, Scopus, así como otras fuentes de literatura gris para las publicaciones sobre ACV y covid-19. Se identificaron datos demográficos, tiempo de aparición del ACV desde el diagnóstico de covid-19. Principales hallazgos radiológicos, laboratorios y pronóstico. RESULTADOS: Se obtuvieron ocho estudios, con 43 sujetos que tuvieron ACV isquémico e infección por SARS-CoV-2. La edad promedio fue de 67,4 años, siendo en su mayoría hombres (58,1%).Un hallazgo importante fue el número de casos de ACV con oclusión de vaso grande en 22 de 31 casos reportados (71%). La mediana de NIHSS fue de 14,5 puntos. Se presentó una mortalidad del 27,5% de los sujetos con ACV El estadio más frecuente por covid-19 fue el de condición severa 58,3%. La aparición del ACV luego de la infección por SARS-CoV-2 fue de 10,6 días en promedio. En los laboratorios se identificó una elevación del fibrinógeno (92%), dímero-D (76%) y LDH (82%) respectivamente. El tratamiento recibido de forma más frecuente para el ACV fue la antiagregación, en 51%, mientras que las terapias de reperfusión se hicieron en el 30% de los casos. La mayoría de los pacientes (93%) presentaron síntomas de covid-19, solo 3 pacientes (7%) no presentaron síntomas típicos de esta enfermedad, sin embargo tuvieron alteración del estado de conciencia asociado al ACV CONCLUSIÓN: Los estados de inflamación e hipercoagulabilidad que se presentan durante la infección por SARS-CoV-2 probablemente están en relación con el desarrollo de ACV, lo cual en este caso podrá explicar el gran número de oclusiones de vaso grande. Los marcadores de inflamación generalmente están presentes. Establecer códigos protegidos de ACV es una medida a efectuar en nuestro medio.

SUMMARY INTRODUCTION: In March 2020, COVID-19 was declared a pandemic by the World Health Organization and cases of stroke related to the virus soon were reported. OBJECTIVES: To become aware of the experiences different parts of the world have encountered with stroke and COVID 19 in order to enhance our knowledge, improve recognition and response to patients in our clinical setting. METHODS: A review of the available observational studies was done using PubMed, Scopus and other sources of gray literature for the publications on stroke and COVID-19. Demographic data, time of stroke onset since the diagnosis of COVID-19, main radiological findings, lab tests and prognosis were identified. RESULTS: Eight studies were selected with 43 subjects who had ischemic stroke and SARS-CoV-2 infection. The average age was 67.4 years, being mostly men (58%). An important finding was the number of stroke cases with large vessel occlusion, with 22 of 31 cases reported (71%). The NIHSS median was 14.5 points, and 27.5% of subjects with stroke and COVID-19 died. The most frequent disease severity for COVID-19 was "severe", accounting for 58% of the cases. The onset of stroke after SARS-CoV-2 infection was 10.6 days on average. In the laboratories an elevation of fibrinogen (92%), D-Dimer (76%) and LDH (82%) were respectively identified. The most frequent treatment received for stroke were antiplatelet medications (51%), while reperfusion therapy was done in 30% of cases. Most patients presented to the hospital with typical symptoms of COVID-19 (93%), (7%) 3 patients did not have respiratory symptoms, however they presented with a decreased level of consciousness associated with stroke findings. CONCLUSION: Inflammation and hypercoagulability, both present during the infection by SARS-CoV-2, are probably related to the development of a stroke, which in this case could explain the large number of large vessel occlusions. Protected stroke code is a protocol that should be implemented in our region.

A Simple High Efficiency Protocol for Pancreatic Islet Isolation from Mice.

Pancreatic islets, also called the Islets of Langerhans, are a cluster of endocrine cells which produces hormones for glucose regulation and other important biological functions. The islets primarily consist of five types of hormone-secreting cells: α cells secrete glucagon, ß cells secrete insulin, δ cells secrete somatostatin, ε cells secrete ghrelin, and PP cells secrete pancreatic polypeptide. Sixty to 80% of the cells in the islets are ß cells, which are the most important cell population to study insulin secretion. Pancreatic islets are a crucial model system to study ex vivo insulin secretion. Acquiring high quality islets is of great importance for diabetes research. Most islet isolation procedures require technically difficult to access site of collagenase injection, harsh and complex digestion procedures, and multiple density gradient purification steps. This paper features a simple high yield mouse islet isolation method with detailed descriptions and realistic demonstrations, showing the following specific steps: 1) injection of collagenase P at the ampulla of Vater, a small area joining the pancreatic duct and the common bile duct, 2) enzymatic digestion and mechanical separation of the exocrine pancreas, and 3) a single gradient purification step. The advantages of this method are the injection of digestive enzyme using the more accessible ampulla of Vater, more complete digestion using combination of enzymatic and mechanical approaches, and a simpler single gradient purification step. This protocol produces approximately 250-350 islets per mouse; and islets are suitable for various ex vivo studies. Possible caveats of this procedure are potentially damaged islets due to enzymatic digestion and/or prolonged gradient incubation, all of which can be largely avoided by careful ad justification of incubation time.

Designed DNA-Encoded IL-36 Gamma Acts as a Potent Molecular Adjuvant Enhancing Zika Synthetic DNA Vaccine-Induced Immunity and Protection in a Lethal Challenge Model.

Identification of novel molecular adjuvants which can boost and enhance vaccine-mediated immunity and provide dose-sparing potential against complex infectious diseases and for immunotherapy in cancer is likely to play a critical role in the next generation of vaccines. Given the number of challenging targets for which no or only partial vaccine options exist, adjuvants that can address some of these concerns are in high demand. Here, we report that a designed truncated Interleukin-36 gamma (IL-36 gamma) encoded plasmid can act as a potent adjuvant for several DNA-encoded vaccine targets including human immunodeficiency virus (HIV), influenza, and Zika in immunization models. We further show that the truncated IL-36 gamma (opt-36γt) plasmid provides improved dose sparing as it boosts immunity to a suboptimal dose of a Zika DNA vaccine, resulting in potent protection against a lethal Zika challenge.

Phosphorylation of Forkhead Protein FoxO1 at S253 Regulates Glucose Homeostasis in Mice.

The transcription factor forkhead box O1 (FoxO1) is a key mediator in the insulin signaling pathway and controls multiple physiological functions, including hepatic glucose production (HGP) and pancreatic ß-cell function. We previously demonstrated that S256 in human FOXO1 (FOXO1-S256), equivalent to S253 in mouse FoxO1 (FoxO1-S253), is a key phosphorylation site mediating the effect of insulin as a target of protein kinase B on suppression of FOXO1 activity and expression of target genes responsible for gluconeogenesis. Here, we investigated the role of FoxO1-S253 phosphorylation in control of glucose homeostasis in vivo by generating global FoxO1-S253A/A knockin mice, in which FoxO1-S253 alleles were replaced with alanine (A substitution) blocking FoxO1-S253 phosphorylation. FoxO1-S253A/A mice displayed mild increases in feeding blood glucose and insulin levels but decreases in fasting blood glucose and glucagon concentrations, as well as a reduction in the ratio of pancreatic α-cells/ß-cells per islet. FoxO1-S253A/A mice exhibited a slight increase in energy expenditure but barely altered food intake and glucose uptake among tissues. Further analyses revealed that FoxO1-S253A/A enhances FoxO1 nuclear localization and promotes the effect of glucagon on HGP. We conclude that dephosphorylation of S253 in FoxO1 may reflect a molecular basis of pancreatic plasticity during the development of insulin resistance.

Inhibitory action of thymol on fecal microbial activity in Tamandua tetradactyla and its effect on glucocorticoid metabolite measurement.

Faecal glucocorticoid measurement is a potentially important tool for improving wildlife conservation, but its use is still limited by methodological issues including the need to avoid modifications of steroids by faecal microorganisms during storage. The freezing of faeces is recommended as a means of avoiding such alterations, but this is costly under non-controlled environmental conditions. The present study was designed to determine whether the application of thymol reduced the proliferation of microorganisms in the faeces of Tamandua tetradactyla and whether it influenced faecal glucocorticoid metabolite (FGM) measurements. Tamandua tetradactyla faeces were individually collected after defaecation, divided into fractions (5.5 g each) and kept in sealed glass Petri dishes at 22 ±â€¯2 °C. A thymol solution (550 µL; 5 mg g-1 feces; 80% ethanol) or an 80% ethanol solution (550 µL, control) was added before storage of faeces. Negative controls for FGM consisted of samples without thymol or ethanol solutions. All samples were evaluated at 0, 24, 48 and 72 h post-defaecation. Thymol was first incubated with a glucocorticoid standard in a faeces-free tube or in a faecal sample in order to determine whether it interfered with FGM measurements. Data showed that thymol did not affect FGM measurements. Post-defaecation time caused a significant reduction in FGM measurements in the negative control, an increment at 48 h in the control, and no change in FGM measurements in thymol treatment. FGM measurements were significantly different between groups (negative control > control - treatment). Thymol caused a significant reduction of up to three orders of magnitude in total coliforms, total aerobic and anaerobic heterotrophic mesophilic bacteria, mold and yeast per gram of faeces at 24, 48 and 72 h. The reduction in microbial activity presumably contributed to the stability of FGM over time. Spore-forming bacteria (SFB) in faeces were not reduced by thymol. We propose thymol as an alternative to freezing since it stabilizes FGMs for at least 3 days after collection in the faeces of Tamandua tetradactyla.

The Birth of Angiotensin: An International Compromise.

Irvine Page in the United States and Eduardo Braun-Menéndez in Argentina led teams of investigators that studied the role of the kidney in blood pressure regulation. Contemporaneously in 1939, each team using different methods discovered and described a new substance now known as angiotensin. At the time of discovery, Page called it "angiotonin" and Braun-Menéndez called it "hipertensina," anglicized to "hypertensin." Over time, the importance of this substance in circulatory control, pathophysiology and pharmacology became indisputable and the need for a single name became obvious. In a remarkable accommodation, Page and Braun-Menéndez agreed to forego any claim to priority and chose a name with elements of both. Following this compromise, Page and Braun-Menéndez went on to become leaders in science in their own countries as well as recognition world-wide while, angiotensin and its derivatives have become standard components in the understanding and treatment of diseases of the heart, kidney and brain.

Lambl's Excrescences: An Enigma of Modern Diagnostic Cardiology.

Lambl's excrescences were first described in 1856 by a Bohemian physician, Vilém Dusan Lambl, and since then have gained widespread attention and controversy within the medical literature. Despite numerous case reports and observational studies, consensus on the significance and management of Lambl's excrescences remains sparse. We describe the case of a 48-year-old male who presented with recurrent embolic strokes. No underlying paroxysmal arrhythmia or inter-atrial shunt was identified, and the only pathological finding was a 1-mm aortic valve strand. We managed this patient successfully using a novel oral anticoagulant.

IL-33 delays metastatic peritoneal cancer progression inducing an allergic microenvironment.

Ovarian cancer is frequently diagnosed as peritoneal carcinomatosis. Unlike other tumor locations, the peritoneal cavity is commonly exposed to gut-breaching and ascending genital microorganisms and has a unique immune environment. IL-33 is a local cytokine that can activate innate and adaptive immunity. We studied the effectiveness of local IL-33 delivery in the treatment of cancer that has metastasized to the peritoneal cavity. Direct peritoneal administration of IL-33 delayed the progression of metastatic peritoneal cancer. Prolongation in survival was not associated with a direct effect of IL-33 on tumor cells, but with major changes in the immune microenvironment of the tumor. IL-33 promoted a significant increase in the leukocyte compartment of the tumor immunoenvironment and an allergic cytokine profile. We observed a substantial increase in the number of activated CD4+ T-cells accompanied by peritoneal eosinophil infiltration, B-cell activation and activation of peritoneal macrophages which displayed tumoricidal capacity. Depletion of CD4+ cells, eosinophils or macrophages reduced the anti-tumor effects of IL-33 but none of these alone were sufficient to completely abrogate its positive benefit. In conclusion, local administration of IL-33 generates an allergic tumor environment resulting in a novel approach for treatment of metastatic peritoneal malignancies, such as advanced ovarian cancer.

Protective Efficacy and Long-Term Immunogenicity in Cynomolgus Macaques by Ebola Virus Glycoprotein Synthetic DNA Vaccines.

Background: There remains an important need for prophylactic anti-Ebola virus vaccine candidates that elicit long-lasting immune responses and can be delivered to vulnerable populations that are unable to receive live-attenuated or viral vector vaccines. Methods: We designed novel synthetic anti-Ebola virus glycoprotein (EBOV-GP) DNA vaccines as a strategy to expand protective breadth against diverse EBOV strains and evaluated the impact of vaccine dosing and route of administration on protection against lethal EBOV-Makona challenge in cynomolgus macaques. Long-term immunogenicity was monitored in nonhuman primates for >1 year, followed by a 12-month boost. Results: Multiple-injection regimens of the EBOV-GP DNA vaccine, delivered by intramuscular administration followed by electroporation, were 100% protective against lethal EBOV-Makona challenge. Impressively, 2 injections of a simple, more tolerable, and dose-sparing intradermal administration followed by electroporation generated strong immunogenicity and was 100% protective against lethal challenge. In parallel, we observed that EBOV-GP DNA vaccination induced long-term immune responses in macaques that were detectable for at least 1 year after final vaccination and generated a strong recall response after the final boost. Conclusions: These data support that this simple intradermal-administered, serology-independent approach is likely important for additional study towards the goal of induction of anti-EBOV immunity in multiple at-risk populations.