Mechanism of Action of Oral Salmonella-Based Vaccine to Prevent and Reverse Type 1 Diabetes in NOD Mice.

A combination therapy of preproinsulin (PPI) and immunomodulators (TGFß+IL10) orally delivered via genetically modified Salmonella and anti-CD3 promoted glucose balance in in NOD mice with recent onset diabetes. The Salmonella bacteria were modified to express the diabetes-associated antigen PPI controlled by a bacterial promoter in conjunction with over-expressed immunomodulating molecules. The possible mechanisms of action of this vaccine to limit autoimmune diabetes remained undefined. In mice, the vaccine prevented and reversed ongoing diabetes. The vaccine-mediated beneficial effects were associated with increased numbers of antigen-specific CD4+CD25+Foxp3+ Tregs, CD4+CD49b+LAG3+ Tr1-cells, and tolerogenic dendritic-cells (tol-DCs) in the spleens and lymphatic organs of treated mice. Despite this, the immune response to Salmonella infection was not altered. Furthermore, the vaccine effects were associated with a reduction in islet-infiltrating lymphocytes and an increase in the islet beta-cell mass. This was associated with increased serum levels of the tolerogenic cytokines (IL10, IL2, and IL13) and chemokine ligand 2 (CCL2) and decreased levels of inflammatory cytokines (IFNγ, GM-CSF, IL6, IL12, and TNFα) and chemokines (CXCL1, CXCL2, and CXCL5). Overall, the data suggest that the Salmonella-based vaccine modulates the immune response, reduces inflammation, and promotes tolerance specifically to an antigen involved in autoimmune diabetes.

A novel approach to determine the critical survival threshold of cellular oxygen within spheroids via integrating live/dead cell imaging with oxygen modeling.

Defining the oxygen level that induces cell death within 3-D tissues is vital for understanding tissue hypoxia; however, obtaining accurate measurements has been technically challenging. In this study, we introduce a noninvasive, high-throughput methodology to quantify critical survival partial oxygen pressure (pO2) with high spatial resolution within spheroids by using a combination of controlled hypoxic conditions, semiautomated live/dead cell imaging, and computational oxygen modeling. The oxygen-permeable, micropyramid patterned culture plates created a precisely controlled oxygen condition around the individual spheroid. Live/dead cell imaging provided the geometric information of the live/dead boundary within spheroids. Finally, computational oxygen modeling calculated the pO2 at the live/dead boundary within spheroids. As proof of concept, we determined the critical survival pO2 in two types of spheroids: isolated primary pancreatic islets and tumor-derived pseudoislets (2.43 ± 0.08 vs. 0.84 ± 0.04 mmHg), indicating higher hypoxia tolerance in pseudoislets due to their tumorigenic origin. We also applied this method for evaluating graft survival in cell transplantations for diabetes therapy, where hypoxia is a critical barrier to successful transplantation outcomes; thus, designing oxygenation strategies is required. Based on the elucidated critical survival pO2, 100% viability could be maintained in a typically sized primary islet under the tissue pO2 above 14.5 mmHg. This work presents a valuable tool that is potentially instrumental for fundamental hypoxia research. It offers insights into physiological responses to hypoxia among different cell types and may refine translational research in cell therapies.NEW & NOTEWORTHY Our study introduces an innovative combinatory approach for noninvasively determining the critical survival oxygen level of cells within small cell spheroids, which replicates a 3-D tissue environment, by seamlessly integrating three pivotal techniques: cell death induction under controlled oxygen conditions, semiautomated imaging that precisely identifies live/dead cells, and computational modeling of oxygen distribution. Notably, our method ensures high-throughput analysis applicable to various cell types, offering a versatile solution for researchers in diverse fields.

A high burden of diabetes and ankle brachial index abnormalities exists in Mexican Americans in South Texas.

Ethnic differences exist in the United States in the interrelated problems of diabetes (DM), peripheral arterial disease (PAD), and leg amputations. The purpose of this study was to determine the prevalence and risk factor associations for subclinical PAD in a population sample of Mexican Americans using the ankle brachial (ABI) index. The ABI-High (higher of the two ankle pressures/highest brachial pressure) and ABI-Low (lower of the two ankle pressures/highest brachial pressure) were calculated to define PAD. Toe brachial index (TBI) was also calculated. 746 participants were included with an age of 53.4 ± 0.9 years, 28.3 % had diabetes mellitus (DM), 12.6 % were smokers, and 51.2 % had hypertension (HTN). Using ABI-High ≤ 0.9, the prevalence of PAD was 2.7 %. This rose to 12.7 % when an ABI-Low ≤ 0.9 was used; 4.0 % of the population had an ABI-High > 1.4. The prevalence of TBI < 0.7 was 3.9 %. DM was a significant risk factor for ABI-High ≤ 0.9 and ABI-High > 1.4, and TBI < 0.7. Increased age, HTN, smoking was associated with ABI-High ≤ 0.9, while being male was associated with ABI-High > 1.4. Increased age, smoking, and lower education were all associated with abnormal TBI. Despite relatively younger mean age than other studied Hispanic cohorts, the present population has a high burden of ABI abnormalities. DM was a consistent risk factor for PAD. These abnormalities indicate an important underlying substrate of vascular and metabolic disease that may predispose this population to the development of symptomatic PAD and incident amputations.

A scalable human islet 3D-culture platform maintains cell mass and function long-term for transplantation.

Present-day islet culture methods provide short-term maintenance of cell viability and function, limiting access to islet transplantation. Attempts to lengthen culture intervals remain unsuccessful. A new method was developed to permit the long-term culture of islets. Human islets were embedded in polysaccharide 3D-hydrogel in cell culture inserts or gas-permeable chambers with serum-free CMRL 1066 supplemented media for up to 8 weeks. The long-term cultured islets maintained better morphology, cell mass, and viability at 4 weeks than islets in conventional suspension culture. In fact, islets cultured in the 3D-hydrogel retained ß cell mass and function on par with freshly isolated islets in vitro and, when transplanted into diabetic mice, restored glucose balance similar to fresh islets. Using gas-permeable chambers, the 3D-hydrogel culture method was scaled up over 10-fold and maintained islet viability and function, although the cell mass recovery rate was 50%. Additional optimization of scale-up methods continues. If successful, this technology could afford flexibility and expand access to islet transplantation, especially single-donor islet-after-kidney transplantation.

One-Step Automatic Radiosynthesis and Evaluation of [18F]TM-30089 as GPR44 Radiotracer.

Recently, a G-protein coupled receptor 44 (GPR44) was discovered to play a significant role in the process of inflammation-related diseases, including cancer and diabetes. However, the precise role of GPR44 has yet to be fully elucidated. Currently, there is a strong and urgent need for the development of GPR44 radiotracers as a non-invasive methodology to explore the exact mechanism of GPR44 on inflammation-related diseases and monitor the progress of therapy. TM-30089 is a potent GPR44 antagonist that exhibits a high specificity and selectivity for GPR44. Its structure contains a fluorine nuclide, which could potentially be replaced with 18F. In the present study, we successfully took a highly effective synthesis strategy that pretreated the unprotected carboxylic acid group of the precursor and developed a feasible one-step automatic radiosynthesis strategy for [18F]TM-30089 with a high radiochemical purity and a good radiochemical yield. We further evaluated this radiotracer using mice models implanted with 1.1 B4 cell lines (GPR44-enriched cell lines) and human islets (high GPR44 expression), respectively. The results revealed the persistent and specific uptake of [18F]TM-30089 in GPR44 region, indicating that [18F]TM-30089 is a promising candidate for targeting GPR44. Further evaluation is ongoing.

Ubiquitous Luciferase Expression in "Firefly Rats" Does Not Alter the Pancreatic Islet Morphology, Metabolism, and Function.

"Firefly rats" ubiquitously express the luciferase reporter gene under the control of constitutively active ROSA26 promoter in inbred Lewis rats. Due to the minimal immunogenicity of luciferase, wide applications of Firefly rats have been reported in solid organ/cell transplantation studies for in vivo imaging, permitting quantitative and non-invasive tracking of the transplanted graft. ROSA26 is a non-coding gene and generally does not affect the expression of other endogenous genes. However, the effect of ubiquitous luciferase expression on islet morphology and function has not been thoroughly investigated, which is critical for the use of Firefly rats as islet donors in islet transplantation studies. Accordingly, in vivo glucose homeostasis (i.e., islet function in the native pancreas) was compared between age-matched luciferase-expressing Firefly rats and non-luciferase-expressing rats. In vivo assessments demonstrated no statistical difference between these rats in non-fasting blood glucose levels, intraperitoneal glucose tolerance tests, and glucose-stimulated serum C-peptide levels. Furthermore, islets were isolated from both rats to compare the morphology, function, and metabolism in vitro. Isolated islets from both rats exhibited similar in vitro characteristics in post-isolation islet yield, islet size, beta cell populations, insulin content per islet, oxygen consumption rate, and glucose-stimulated insulin secretion. In conclusion, ubiquitous luciferase expression in Firefly rats does not affect their islet morphology, metabolism, and function; this finding is critical and enables the use of isolated islets from Firefly rats for the dual assessment of islet graft function and bioluminescence imaging of islet grafts.

Detection of bipolar disorder in the prodromal phase: A systematic review of assessment instruments.

BACKGROUND: Early detection of prodromal symptoms may contribute to improving the prognosis of patients with bipolar disorder (BD). The main objective of this systematic review is to present the different procedures for the identification of initial and relapse prodromes in these patients. METHODS: PsycINFO, Web of Science and PubMed databases were searched using a predetermined strategy, until January 4, 2022. Then, by means of a regulated process, studies that used a BD prodrome detection procedure, in English-language and all ages participants were selected. Quantitative and qualitative studies were assessed using a modified version of the Newcastle-Ottawa Scale and by Critical Appraisals Skills Programme checklist, respectively. RESULTS: Forty-five studies were selected. Of these, 26 used procedures for identifying initial prodromes (n = 8014) and 19 used procedures for detecting relapse prodromes (n = 1136). The interview was the most used method in the detection of both types of prodromes (k = 30 papers, n = 4068). It was variable in its degree of structure. Mobile applications and digital technologies are gaining importance in the detection of the relapse prodromes. LIMITATIONS: A retrospective design in most papers, small samples sizes, existence of persistent subsyndromal symptoms and difficulty to identify the end of the prodrome and the onset of the disorder. CONCLUSIONS: There is a wide variety of assessment instruments to detect prodromes in BD, among which the clinical interview is most frequently used. Future research should consider development of a brief tool to be applied in different formats to patients and family members.

Emotional processing in patients with single brain damage in the right hemisphere.

BACKGROUND: The interest in the relationship between brain damage and social cognition has increased in recent years. The objectives of the present study were the following: (1) to evaluate and compare emotional facial recognition and subjective emotional experience in patients who have suffered a single ischemic stroke in the right hemisphere (RH) and in healthy people, (2) to analyze the relationship between both variables in both groups of subjects, and (3) to analyze the association between the cerebral location of the stroke and these two variables. METHODS: Emotional facial recognition and the subjective emotional experience of 41 patients who had suffered a single ischemic stroke in the RH and 45 volunteers without previous cerebrovascular pathology were evaluated. RESULTS: Brain damaged patients performed lower in facial emotional recognition and had a less intense subjective emotional response to social content stimuli compared to healthy subjects. Likewise, among patients with RH ischemic stroke, we observed negative associations between facial recognition of surprise and reactivity to unpleasant images, and positive associations between recognition of disgust and reactivity to pleasant images. Finally, patients with damage in the caudate nucleus of the RH presented a deficit in the recognition of happiness and sadness, and those with damage in the frontal lobe exhibited a deficit in the recognition of surprise, compared to those injured in other brain areas. CONCLUSIONS: Emotional facial recognition and subjective emotional experience are affected in patients who have suffered a single ischemic stroke in the RH. Professionals caring for stroke patients should improve their understanding of the general condition of affected persons and their environment, assess for risk of depression, and facilitate their adaptation to work, family, and social environments.

Micropyramid-patterned, oxygen-permeable bottomed dish for high density culture of pancreatic islets.

The need for maintaining cell-spheroid viability and function within high-density cultures is unmet for various clinical and experimental applications, including cell therapies. One immediate application is for transplantation of pancreatic islets, a clinically recognized treatment option to cure type 1 diabetes; islets are isolated from a donor for subsequent culture prior to transplantation. However, high seeding conditions cause unsolicited fusion of multiple spheroids, thereby limiting oxygen diffusion to induce hypoxic cell death. Here we introduce a culture dish incorporating a micropyramid-patterned surface to prevent the unsolicited fusion and oxygen-permeable bottom for optimal oxygen environment. A 400µm-thick, oxygen-permeable polydimethylsiloxane sheet topped with micropyramid pattern of 400µm-base and 200µm-height was fabricated to apply to the 24-well plate format. The micropyramid pattern separated the individual pancreatic islets to prevent the fusion of multiple islets. This platform supported the high oxygen demand of islets at high seeding density at 260 islet equivalents cm-2, a 2-3-fold higher seeding density compared to the conventional islet culture used in a preparation for the clinical islet transplantations, demonstrating improved islet morphology, metabolism and function in a 4 d-culture. Transplantation of these islets into immunodeficient diabetic mice exhibited significantly improved engraftment to achieve euglycemia compared to islets cultured in the conventional culture wells. Collectively, this simple design modification allows for high-density cultures of three-dimensional cell spheroids to improve the viability and function for an array of investigational and clinical replacement tissues.

Submilligram Level of Beetle Antifreeze Proteins Minimize Cold-Induced Cell Swelling and Promote Cell Survival.

Hypothermic (cold) preservation is a limiting factor for successful cell and tissue transplantation where cell swelling (edema) usually develops, impairing cell function. University of Wisconsin (UW) solution, a standard cold preservation solution, contains effective components to suppress hypothermia-induced cell swelling. Antifreeze proteins (AFPs) found in many cold-adapted organisms can prevent cold injury of the organisms. Here, the effects of a beetle AFP from Dendroides canadensis (DAFP-1) on pancreatic ß-cells preservation were first investigated. As low as 500 µg/mL, DAFP-1 significantly minimized INS-1 cell swelling and subsequent cell death during 4 °C preservation in UW solution for up to three days. However, such significant cytoprotection was not observed by an AFP from Tenebrio molitor (TmAFP), a structural homologue to DAFP-1 but lacking arginine, at the same levels. The cytoprotective effect of DAFP-1 was further validated with the primary ß-cells in the isolated rat pancreatic islets in UW solution. The submilligram level supplement of DAFP-1 to UW solution significantly increased the islet mass recovery after three days of cold preservation followed by rewarming. The protective effects of DAFP-1 in UW solution were discussed at a molecular level. The results indicate the potential of DAFP-1 to enhance cell survival during extended cold preservation.

Efficacy of Smartphone Apps in Patients With Depressive Disorders: A Systematic Review.

Background: Digital interventions have become an accessible format in clinical practice to provide better support for patients with mental disorders. However, the clinical efficacy in patients with depressive disorders is not well known. We aimed to determine the efficacy of smartphone applications (apps) in patients diagnosed with a depressive disorder. Method: An electronic database search was performed of PubMed, PsycINFO, and Web of Science, to identify relevant articles up to June 12, 2021. Peer-reviewed articles were screened and selected based on predetermined inclusion and exclusion criteria. Results: Seven articles met the inclusion criteria and therefore were selected for the systematic review, which included a total of 651 patients. The results were heterogeneous, essentially due to the different methodologies used in the selected studies. Conclusions: Digital smartphone-delivered interventions do not appear to reduce depressive symptomatology nor improve the quality of life in patients diagnosed with depressive disorders when compared to an active control group. Taking into account the inherent methodological difficulties and the variability among such studies, it is apparent that further research-with more methodologically refined clinical trials, including larger sample sizes-is needed.

Critical Considerations in Bioluminescence Imaging of Transplanted Islets: Dynamic Signal Change in Early Posttransplant Phase and Signal Absorption by Tissues.

OBJECTIVES: In pancreatic islet transplantation studies, bioluminescence imaging enables quantitative and noninvasive tracking of graft survival. Amid the recent heightened interest in extrahepatic sites for islet and stem cell-derived beta-like cell transplantations, proper understanding the nature of bioluminescence imaging in these sites is important. METHODS: Islets isolated from Firefly rats ubiquitously expressing luciferase reporter gene in Lewis rats were transplanted into subcutaneous or kidney capsule sites of wild-type Lewis rats or immunodeficient mice. Posttransplant changes of bioluminescence signal curves and absorption of bioluminescence signal in transplantation sites were examined. RESULTS: The bioluminescence signal curve dynamically changed in the early posttransplantation phase; the signal was low within the first 5 days after transplantation. A substantial amount of bioluminescence signal was absorbed by tissues surrounding islet grafts, correlating to the depth of the transplanted site from the skin surface. Grafts in kidney capsules were harder to image than those in the subcutaneous site. Within the kidney capsule, locations that minimized depth from the skin surface improved the graft detectability. CONCLUSIONS: Posttransplant phase and graft location/depth critically impact the bioluminescence images captured in islet transplantation studies. Understanding these parameters is critical for reducing experimental biases and proper interpretation of data.

Single and Combined Effect of Mild-Heat Treatment and Alginate Coatings on Quality Preservation of Minimally Processed Bunching Green Onions.

Bunching green onion is an Allium species that has been widely used in food flavorings and seasonings. This vegetable experiences a rapid loss of quality during storage due to physiological changes and microbial spoilage. In the current work, the single and combined effect of mild-heat treatment (55 °C for 60 s) and alginate edible coatings on the quality preservation of minimally processed bunching green onions was studied. Control and treated samples were stored at 4 °C for 15 days and examined periodically in terms of their respiration rate, weight loss, pH, soluble solids content, firmness, total polyphenol content, antioxidant activity, microbial count, decay ratio, and overall visual quality. The results showed that the combination of mild heat and alginate edible coatings was the most effective approach to slow down the respiration rate and the incidence of decay in the minimally processed bunching green onions. In addition, the treatments with alginate coating alone or combined with mild-heat treatment showed the best performance for maintaining the overall visual quality of the products during the storage.

Descripción de pacientes con displasia del desarrollo de cadera en Hospital Clínico Herminda Martin, Chillán, entre los años 2020-2021 / Description of patients with developmental dysplasia of the hip at the Hospital Clínico Herminda Martin, Chillán, between the years 2020-2021

Introducción: La displasia del desarrollo de la cadera (DDC), una patología multifactorial más prevalente en el sexo femenino, afecta a lactantes, y el diagnóstico y tratamiento oportuno permiten evitar consecuencias posteriores, morbimortalidad significativa y una carga en salud importante. Objetivo: Conocer la realidad local en relación con el diagnóstico y tratamiento de la DDC, describiendo demográficamente a los pacientes evaluados por médicos especialistas, específicamente traumatólogos infantiles del Hospital Clínico Herminda Martin (HCHM) en Chillán. Materiales y métodos: Se llevó a cabo un estudio descriptivo transversal de una cohorte de pacientes evaluados mediante radiografías en el HCHM por sospecha de DDC, entre junio de 2020 y julio de 2021. Se recopilaron variables como sexo, tipo de compromiso del desarrollo de la cadera y tratamiento utilizado. Resultados: De 146 pacientes evaluados, el 83.6% correspondían al sexo femenino, mientras que el 16.4% eran del sexo masculino. En cuanto a los tratamientos ortopédicos, el 82.7% fueron mujeres y el 17.2% hombres, siendo las correas de Pavlik el tratamiento más utilizado. Solo 7 pacientes requirieron tratamiento quirúrgico. Discusión: Los resultados obtenidos se respaldan en la evidencia internacional, reflejando realidades similares a la situación local. La relevancia de este estudio radica en la falta de una base de datos nacional o local actualizada sobre la patología, sumado a la ventaja de informar acerca de los tratamientos disponibles y la adherencia a ellos. Esto permite reflejar el comportamiento de la población local durante los años 2020-2021. Las limitaciones incluyen el seguimiento, debido a restricciones por la pandemia y la falta de datos previos.

Introduction: Developmental dysplasia of the hip (DDH), a multifactorial pathology more prevalent in females, affects infants, where timely diagnosis and treatment avoid subsequent consequences, significant morbidity, and mortality, as well as the health burden it generates. Objective: To understand the local reality regarding the diagnosis and treatment of DDH by describing the demographic characteristics of patients evaluated by medical specialists, specifically child traumatologists from the Hospital Clínico Herminda Martin (HCHM) in Chillán. Materials and Methods: A cross-sectional descriptive study was conducted on a cohort of patients evaluated through radiographs at HCHM due to suspected DDH between June 2020 and July 2021. Variables collected included sex, type of hip development compromise, and treatment used. Results: Out of 146 patients, 83.6% were female and 16.4% were male. In terms of orthopedic treatments, 82.7% were females, and 17.2% were males, with Pavlik straps being the most used orthopedic treatment. Only 7 patients underwent surgical treatment. Discussion: The obtained results align with international evidence, reflecting realities similar to the local situation. The study's significance lies in the absence of an updated national or local database on the pathology. Additionally, it provides insights into available treatments and patient adherence, offering a snapshot of the local population's behavior during 2020-2021. Limitations include patient follow-up challenges due to pandemic restrictions and the lack of pre-existing data.

A Multiparametric Assessment of Human Islets Predicts Transplant Outcomes in Diabetic Mice.

Prior to transplantation into individuals with type 1 diabetes, in vitro assays are used to evaluate the quality, function and survival of isolated human islets. In addition to the assessments of these parameters in islet, they can be evaluated by multiparametric morphological scoring (0-10 points) and grading (A, B, C, D, and F) based on islet characteristics (shape, border, integrity, single cells, and diameter). However, correlation between the multiparametric assessment and transplantation outcome has not been fully elucidated. In this study, 55 human islet isolations were scored using this multiparametric assessment. The results were correlated with outcomes after transplantation into immunodeficient diabetic mice. In addition, the multiparametric assessment was compared with oxygen consumption rate of isolated islets as a potential prediction factor for successful transplantations. All islet batches were assessed and found to score: 9 points (n = 18, Grade A), 8 points (n = 19, Grade B), and 7 points (n = 18, Grade B). Islets that scored 9 (Grade A), scored 8 (Grade B) and scored 7 (Grade B) were transplanted into NOD/SCID mice and reversed diabetes in 81.2%, 59.4%, and 33.3% of animals, respectively (P < 0.0001). Islet scoring and grading correlated well with glycemic control post-transplantation (P < 0.0001) and reversal rate of diabetes (P < 0.05). Notably, islet scoring and grading showed stronger correlation with transplantation outcome compared to oxygen consumption rate. Taken together, a multiparametric assessment of isolated human islets was highly predictive of transplantation outcome in diabetic mice.

Spanish Versions of the Outcome Rating Scale and the Session Rating Scale: Normative Data, Reliability, and Validity.

Routine outcome monitoring (ROM) uses standardized measures to both track and inform mental health service delivery. Use of ROM has been shown to improve the outcome of psychotherapy when applied to different types of patients. The present research was designed to determine the reliability and validity of the Outcome Rating Scale (ORS) and the Session Rating Scale (SRS) in a sample of Spanish patients. After a controlled process of translation into the Spanish that is spoken and written in Spain (i.e., in Europe, as distinct from, e.g., Latin American Spanish), both measures were completed by patients of an outpatient mental health unit during eight sessions of psychotherapy. Sixty mental health patients filled out the ORS and 59 the SRS. In addition, the ORS was completed by 33 people who constituted the non-clinical sample. The cut-off of the ORS was 24.52 points, and the Reliable Change Index (RCI) was 9.15 points. ORS and SRS scores exhibited excellent internal consistency. The temporal stability of the SRS was adequate. The convergent and discriminant validity of the two measures were adequate. Regarding the factorial validity of the ORS and the SRS, in the third psychotherapy session, confirmatory factor analyses evidenced the existence of a unifactorial model. The predictive validity of SRS was acceptable. The ORS was sensitive to changes in patients' symptoms. In conclusion, compared to the original English versions of the ORS and SRS measures, the Spanish versions of the measures are also reliable and valid.

Emergence of Coupled Rotor Dynamics in Metal-Organic Frameworks via Tuned Steric Interactions.

The organic components in metal-organic frameworks (MOFs) are unique: they are embedded in a crystalline lattice, yet, as they are separated from each other by tunable free space, a large variety of dynamic behavior can emerge. These rotational dynamics of the organic linkers are especially important due to their influence over properties such as gas adsorption and kinetics of guest release. To fully exploit linker rotation, such as in the form of molecular machines, it is necessary to engineer correlated linker dynamics to achieve their cooperative functional motion. Here, we show that for MIL-53, a topology with closely spaced rotors, the phenylene functionalization allows researchers to tune the rotors' steric environment, shifting linker rotation from completely static to rapid motions at frequencies above 100 MHz. For steric interactions that start to inhibit independent rotor motion, we identify for the first time the emergence of coupled rotation modes in linker dynamics. These findings pave the way for function-specific engineering of gear-like cooperative motion in MOFs.

Seven years on from the Canadian Airway Focus Group Difficult Airway Guidelines: an observational survey.

PURPOSE: Numerous clinical guidelines are available for management of the unanticipated difficult airway. It is unclear if practice recommendations are endorsed on regional, local, or individual levels. The objective of this observational study was to examine local and regional use of airway guidelines by anesthesiologists within a hospital partnership in Southwestern Ontario. METHODS: Using a paper survey, distributed locally to consultant and trainee anesthesiologists in a tertiary hospital, we examined individual clinical and educational practices regarding guideline use in airway management. Respondents were asked to report which published guideline they used for unanticipated airway difficulty. The effectiveness of dissemination of the national Canadian airway guidelines-the Canadian Airway Focus Group (CAFG) Difficult Airway Guidelines-was examined. We invited anesthesia department heads within the regional hospital partnership to complete an electronic survey investigating departmental adoption of guidelines. RESULTS: The response rate was 70% locally (79/112 anesthesiologists) and 52% regionally (11/21 department heads). Approximately 80% (64/79) of respondents reported using a formal clinical guideline if unanticipated difficulty with airway management was encountered. Seventy-two per cent of respondents (57/79) were aware of the published CAFG guidelines. Approximately 30% (16/51) of consultant anesthesiologists reported using the CAFG guidelines in clinical practice. Within the hospital partnership, 36% (4/11) of departments formally endorsed use of a specific airway management guideline. CONCLUSIONS: Despite widespread awareness of the national CAFG Difficult Airway Guidelines, they are not widely adopted in clinical practice. Further research is warranted to explore barriers to adoption of airway management guidelines for both individual anesthesiologists and anesthesia departments.

RéSUMé: OBJECTIF: De nombreuses lignes directrices cliniques sont disponibles pour la prise en charge des voies aériennes difficiles non anticipées. Nous ne savons pas si ces recommandations de pratique sont suivies aux niveaux régional, local ou individuel. L'objectif de cette étude observationnelle était d'examiner l'utilisation locale et régionale des lignes directrices sur les voies aériennes par des anesthésiologistes provenant d'un partenariat hospitalier dans le sud-ouest de l'Ontario. MéTHODE: À l'aide d'une enquête papier, distribuée localement aux anesthésiologistes et aux résidents en anesthésiologie d'un hôpital de soins tertiaires, nous avons examiné les pratiques cliniques et éducatives individuelles concernant l'utilisation des lignes directrices dans la prise en charge des voies aériennes. On a demandé aux répondants de divulguer les lignes directrices publiées qu'ils utilisaient lorsqu'ils étaient confrontés à des difficultés imprévues au niveau des voies aériennes. L'efficacité de diffusion des lignes directrices nationales canadiennes sur les voies aériennes­les Lignes directrices sur les voies aériennes difficiles du Canadian Airway Focus Group (CAFG)­a été examinée. Nous avons invité les chefs des départements d'anesthésiologie au sein du partenariat hospitalier régional à remplir un sondage électronique sur l'adoption des lignes directrices par leur département. RéSULTATS: Le taux de réponse était de 70 % au niveau local (79/112) et de 52 % au niveau régional (11/21 chefs de département). Environ 80 % (64/79) des répondants ont déclaré avoir utilisé des lignes directrices cliniques officielles en cas de difficultés imprévues dans la prise en charge des voies aériennes. Soixante-douze pour cent des répondants (57/79) avaient connaissance des lignes directrices publiées par le CAFG. Environ 30 % (16/51) des anesthésiologistes ont déclaré avoir utilisé les lignes directrices du CAFG dans leur pratique clinique. Au sein du partenariat hospitalier, 36 % (4/11) des départements ont officiellement appuyé l'utilisation d'une ligne directrice spécifique pour la prise en charge des voies aériennes. CONCLUSION: Malgré une importante sensibilisation aux Lignes directrices nationales sur les voies aériennes difficiles du CAFG, ces dernières ne sont pas largement adoptées dans la pratique clinique. D'autres recherches sont nécessaires pour explorer les obstacles à l'adoption de lignes directrices pour la prise en charge des voies aériennes tant par les anesthésiologistes que par les départements d'anesthésie.

Oral Salmonella msbB Mutant as a Carrier for a Salmonella-Based Vaccine for Prevention and Reversal of Type 1 Diabetes.

A therapy that includes an oral vaccine for type 1 diabetes (T1D) using live attenuated Salmonella MvP728 (ΔhtrA/ΔpurD), cytokines (IL10 and TGFß) and preproinsulin (PPI) antigen in combination with a sub-therapeutic dose of anti-CD3 mAb was developed by our team. The vaccine combination therapy reduced insulitis and prevented and reversed diabetes in non-obese diabetic (NOD) mice. Here, we show the effectiveness of an alternative Salmonella mutant (ΔmsbB) as a carrier strain, which is anticipated to have lower risks of an inflammatory response and septicemia as a result of modification in the lipopolysaccharide (LPS) via detoxification of lipid A. This mutant strain proved to have highly reduced pathogenic side effects. Salmonella strain ΔmsbB expressed autoantigens and in combination with cytokines and anti-CD3 mAb, successfully prevented and reversed T1D to levels comparable to the previously used carrier strain ΔhtrA/ΔpurD. Additionally, the Salmonella msbB mutant resulted in higher rates of host cell infection. These results further demonstrate the potential of an oral Salmonella-based combined therapy in the treatment of early T1D.