IRF8-driven reprogramming of the immune microenvironment enhances anti-tumor adaptive immunity and reduces immunosuppression in murine glioblastoma.

BACKGROUND: Glioblastoma (GBM) has a highly immunosuppressive tumor immune microenvironment (TIME), largely mediated by myeloid-derived suppressor cells (MDSCs). Here, we utilized a retroviral replicating vector (RRV) to deliver Interferon Regulatory Factor 8 (IRF8), a master regulator of type 1 conventional dendritic cell (cDC1) development, in a syngeneic murine GBM model. We hypothesized that RRV-mediated delivery of IRF8 could "reprogram" intratumoral MDSCs into antigen-presenting cells (APCs) and thereby restore T-cell responses. METHODS: Effects of RRV-IRF8 on survival and tumor growth kinetics were examined in the SB28 murine GBM model. Immunophenotype was analyzed by flow cytometry and gene expression assays. We assayed functional immunosuppression and antigen presentation by ex vivo T-cell-myeloid co-culture. RESULTS: Intratumoral injection of RRV-IRF8 in mice bearing intracerebral SB28 glioma significantly suppressed the tumor growth and prolonged survival. RRV-IRF8 treated tumors exhibited significant enrichment of cDC1s and CD8+ T-cells. Additionally, myeloid cells derived from RRV-IRF8 tumors showed decreased expression of the immunosuppressive markers Arg1 and IDO1 and demonstrated reduced suppression of naïve T-cell proliferation in ex vivo co-culture, compared to controls. Furthermore, DCs from RRV-IRF8 tumors showed increased antigen presentation compared to those from control tumors. In vivo treatment with azidothymidine (AZT), a viral replication inhibitor, showed that IRF8 transduction in both tumor and non-tumor cells is necessary for survival benefit, associated with a reprogrammed, cDC1- and CD8 T-cell-enriched TIME. CONCLUSIONS: Our results indicate that reprogramming of glioma-infiltrating myeloid cells by in vivo expression of IRF8 may reduce immunosuppression and enhance antigen presentation, achieving improved tumor control.

IRF8-driven reprogramming of the immune microenvironment enhances anti-tumor adaptive immunity and reduces immunosuppression in murine glioblastoma.

Background: Glioblastoma (GBM) has a highly immunosuppressive tumor immune microenvironment (TIME), largely mediated by myeloid-derived suppressor cells (MDSCs). Here, we utilized a retroviral replicating vector (RRV) to deliver Interferon Regulatory Factor 8 (IRF8), a master regulator of type 1 conventional dendritic cell (cDC1) development, in a syngeneic murine GBM model. We hypothesized that RRV-mediated delivery of IRF8 could "reprogram" intratumoral MDSCs into antigen-presenting cells (APCs) and thereby restore T-cell responses. Methods: Effects of RRV-IRF8 on survival and tumor growth kinetics were examined in the SB28 murine GBM model. Immunophenotype was analyzed by flow cytometry and gene expression assays. We assayed functional immunosuppression and antigen presentation by ex vivo T-cell-myeloid co-culture. Results: Mice with RRV-IRF8 pre-transduced intracerebral tumors had significantly longer survival and slower tumor growth compared to controls. RRV-IRF8 treated tumors exhibited significant enrichment of cDC1s and CD8+ T-cells. Additionally, myeloid cells derived from RRV-IRF8 tumors showed decreased expression of the immunosuppressive markers Arg1 and IDO1 and demonstrated reduced suppression of naïve T-cell proliferation in ex vivo co-culture, compared to controls. Furthermore, DCs from RRV-IRF8 tumors showed increased antigen presentation compared to those from control tumors. In vivo treatment with azidothymidine (AZT), a viral replication inhibitor, showed that IRF8 transduction in both tumor and non-tumor cells is necessary for survival benefit, associated with a reprogrammed, cDC1- and CD8 T-cell-enriched TIME. Conclusions: Our results indicate that reprogramming of glioma-infiltrating myeloid cells by in vivo expression of IRF8 may reduce immunosuppression and enhance antigen presentation, achieving improved tumor control.

The Use of MRI to Detect Malignancy in a Patient Presenting With Unilateral Bloody Nipple Discharge.

Nipple discharge presents as either physiological, which is green, white, or yellow, or is considered pathological, which is typically unilateral, spontaneous, and bloody. Bloody nipple discharge (BND) can be due to underlying malignancy or premalignant lesions. Mammogram (MMG), ultrasound (US), MRI, and ductography are all used to evaluate BND, but different modalities offer greater value in the diagnostic process. Here, we present a case that demonstrates the ability of MRI to detect abnormalities not seen on MMG and US in the setting of BND due to underlying malignancy. The use of MRI earlier in the diagnostic process allows for the use of breast-conserving measures and decreases the possibility of metastasis. This would result in less of a need for more aggressive treatments.

SARS-CoV-2 Spike triggers barrier dysfunction and vascular leak via integrins and TGF-ß signaling.

Severe COVID-19 is associated with epithelial and endothelial barrier dysfunction within the lung as well as in distal organs. While it is appreciated that an exaggerated inflammatory response is associated with barrier dysfunction, the triggers of vascular leak are unclear. Here, we report that cell-intrinsic interactions between the Spike (S) glycoprotein of SARS-CoV-2 and epithelial/endothelial cells are sufficient to induce barrier dysfunction in vitro and vascular leak in vivo, independently of viral replication and the ACE2 receptor. We identify an S-triggered transcriptional response associated with extracellular matrix reorganization and TGF-ß signaling. Using genetic knockouts and specific inhibitors, we demonstrate that glycosaminoglycans, integrins, and the TGF-ß signaling axis are required for S-mediated barrier dysfunction. Notably, we show that SARS-CoV-2 infection caused leak in vivo, which was reduced by inhibiting integrins. Our findings offer mechanistic insight into SARS-CoV-2-triggered vascular leak, providing a starting point for development of therapies targeting COVID-19.

Sulfated ß-glucan from Agaricus subrufescens inhibits flavivirus infection and nonstructural protein 1-mediated pathogenesis.

Despite substantial morbidity and mortality, no therapeutic agents exist for treatment of dengue or Zika, and the currently available dengue vaccine is only recommended for dengue virus (DENV)-immune individuals. Thus, development of therapeutic and/or preventive drugs is urgently needed. DENV and Zika virus (ZIKV) nonstructural protein 1 (NS1) can directly trigger endothelial barrier dysfunction and induce inflammatory responses, contributing to vascular leak in vivo. Here we evaluated the efficacy of the (1-6,1-3)-ß-D-glucan isolated from Agaricus subrufescens fruiting bodies (FR) and its sulfated derivative (FR-S) against DENV-2 and ZIKV infection and NS1-mediated pathogenesis. FR-S, but not FR, significantly inhibited DENV-2 and ZIKV replication in human monocytic cells (EC50 = 36.5 and 188.7 µg/mL, respectively) when added simultaneously with viral infection. No inhibitory effect was observed when FR or FR-S were added post-infection, suggesting inhibition of viral entry as a mechanism of action. In an in vitro model of endothelial permeability using human pulmonary microvascular endothelial cells (HPMECs), FR and FR-S (0.12 µg/mL) inhibited DENV-2 NS1- and ZIKV NS1-induced hyperpermeability by 50% and 100%, respectively, as measured by Trans-Endothelial Electrical Resistance. Treatment with 0.25 µg/mL of FR and FR-S inhibited DENV-2 NS1 binding to HPMECs. Further, FR-S significantly reduced intradermal hyperpermeability induced by DENV-2 NS1 in C57BL/6 mice and protected against DENV-induced morbidity and mortality in a murine model of dengue vascular leak syndrome. Thus, we demonstrate efficacy of FR-S against DENV and ZIKV infection and NS1-induced endothelial permeability in vitro and in vivo. These findings encourage further exploration of FR-S and other glycan candidates for flavivirus treatment alone or in combination with compounds with different mechanisms of action.

Open source timed pressure control hardware and software for delivery of air mediated distensions in animal models.

Studying the visceral sensory component of peripheral nervous systems can be challenging due to limited options for consistent and controlled stimulation. One method for mechanical stimulation of hollow organs, including the colon and bladder, is controlled distensions mediated by compressed air. For example, distension of the bladder can be used as an assay for bladder nociception. Bladder distension causes a corresponding increase in abdominal electromyography, which increases with distension pressure and is attenuated with analgesics. However, the hardware used to control these distensions are primarily all one-off custom builds, without clear directions on how to build your own. This has made it difficult for these methods to be fully utilized and replicated as not everyone has the access, knowledge, and resources required to build this controller. Here we show an open-source Arduino-based system for controlling a solenoid valve to deliver timed pressure distensions in the experimental model. This device can be controlled by one of two methods through direct TTL pulses from the experimenter's data acquisition software (ex. CED Spike2) or by a graphical user interface, where the user can set the time before, during, and after distension as well as the number of cycles. This system's low cost and relative ease to build will allow more groups to utilize timed pressure distensions in their experiments.

SARS-CoV-2 Spike triggers barrier dysfunction and vascular leak via integrins and TGF-ß signaling.

Severe COVID-19 is associated with epithelial and endothelial barrier dysfunction within the lung as well as in distal organs. While it is appreciated that an exaggerated inflammatory response is associated with barrier dysfunction, the triggers of this pathology are unclear. Here, we report that cell-intrinsic interactions between the Spike (S) glycoprotein of SARS-CoV-2 and epithelial/endothelial cells are sufficient to trigger barrier dysfunction in vitro and vascular leak in vivo , independently of viral replication and the ACE2 receptor. We identify an S-triggered transcriptional response associated with extracellular matrix reorganization and TGF-ß signaling. Using genetic knockouts and specific inhibitors, we demonstrate that glycosaminoglycans, integrins, and the TGF-ß signaling axis are required for S-mediated barrier dysfunction. Our findings suggest that S interactions with barrier cells are a contributing factor to COVID-19 disease severity and offer mechanistic insight into SARS-CoV-2 triggered vascular leak, providing a starting point for development of therapies targeting COVID-19 pathogenesis.

The role of real-time interactive video consultations in dental practice during the recovery and restoration phase of the COVID-19 outbreak.

The COVID-19 outbreak has caused many challenges for the dental profession and workforce. With the suspension of routine dental services at the end of March 2020, there is a significant number of the general population with acute dental problems. In addition to these, there are patients who had treatment suspended partway through their treatment plans and therefore there is now a backlog of patients awaiting dental care. One challenge that will face the profession moving forward will be facilitating access to these patients and determining which patients should be seen as a priority for necessary dental care. This article looks at the role of real-time interactive video consultations in dental practice during the recovery and restoration phase of the COVID-19 outbreak and how they could facilitate the management of patients waiting to access dental care.

The twin block injection: an adjunctive clinical aid for the management of acute arthrogenous temporomandibular joint dysfunction.

BACKGROUND: The twin block Injection is a novel nerve block that has been shown previously to be efficacious in the management of masticatory myofascial pain. Little is known about its effectiveness for reducing pain from the temporomandibular joint (TMJ). CASE REPORT: A 19-year-old man presented with limited mouth opening with pain in the left side of his face. After a thorough history and examination was completed, the diagnosis was acute anterior disc displacement without reduction in the left TMJ and myalgia of the left side temporalis and masseter muscles. After receiving the twin block injection, the patient reported that the pain in his TMJ had reduced along with the concomitant myalgia. The twin block injection is efficacious for the management of both arthrogenous and myogenous sources of temporomandibular disorders.

Pharmacokinetics of chloramphenicol base in horses and comparison to compounded formulations.

Chloramphenicol is commonly used in horses; however, there are no studies evaluating the pharmacokinetics of veterinary canine-approved tablets. Studies using different formulations and earlier analytical techniques led to concerns over low bioavailability in horses. Safety concerns about human health have led many veterinarians to prescribe compounded formulations that are already in suspension or paste form. The objective of this study was to evaluate the pharmacokinetics of approved chloramphenicol tablets in horses, along with compounded preparations. The hypothesis was that chloramphenicol has low absorption and a short half-life in horses leading to low serum concentrations and that compounded preparations have lower relative bioavailability. Seven horses were administered chloramphenicol tablets (50 mg/kg orally). In a crossover design, they were administered two compounded preparations to compare all three formulations at the same dose (50 mg/kg). Cmax was 5.25 ± 4.07 µg/ml at 4.89 hr, 4.96 ± 3.31 µg/ml at 4.14 hr, and 3.84 ± 2.96 µg/ml at 4.39 hr for the tablets, paste, and suspension, respectively. Elimination half-life was 2.65 ± 0.75, 3.47 ± 1.47, and 4.36 ± 4.54 hr for tablets, paste, and suspension, respectively. The AUC0→∞ was 17.93 ± 7.69, 16.25 ± 1.85, and 14.00 ± 5.47 hr*µg/ml for the tablets, compounded paste, and compounded suspension, respectively. Relative bioavailability of compounded suspension and paste was 78.1% and 90.6%. Cmax after administration of all formulations did not reach the recommended MIC target of 8 µg/ml set by the Clinical Laboratory Standards Institute (CLSI) for most bacteria. Multidose studies are warranted, but the low serum concentrations suggest that bacteria with MIC values lower than CLSI recommendations should be targeted in adult horses.

Drug residues in poultry meat: A literature review of commonly used veterinary antibacterials and anthelmintics used in poultry.

Poultry meat is widely consumed throughout the world and production practices often include the administration of pharmaceutical products. When appropriate, extra-label drug use of medications is necessary, but scientifically derived drug withdrawal intervals must be observed so that poultry meat is not contaminated with drug residues which could pose health risks to consumers. Over the past decade, there has been increased advocacy for judicious use of antimicrobial drugs for treating food animals. Judicious use of medications is commonly referred to as practices that reduce antibiotic resistance, but also includes residue avoidance. In that light, many investigators have performed scientific studies and have published estimated pharmacokinetic parameters for veterinary medications used in commercial avian species. This manuscript is a review of medication classes that have been studied in poultry (mostly chickens) with an emphasis on drug residue depletion in poultry meat.

Acute Bacterial Skin and Skin Structure Infections Treated with Intravenous Antibiotics in the Emergency Department or Observational Unit: Experience at the Detroit Medical Center.

INTRODUCTION: Acute bacterial skin and skin structure infections (ABSSSIs) are frequently treated in emergency departments (EDs) or observation units (OUs) initially with intravenous (IV) antibiotics before discharge on oral therapy. This study aims to describe ABSSSI patients discharged directly from EDs/OUs. METHODS: This is a retrospective cohort study of patients with ABSSSIs treated in EDs/OUs of the Detroit Medical Center from 2012 to 2014. Adults with less than 24 h of IV antibiotics without hospital admission were included. Demographics, clinical characteristics, and severity were compared between ED and OU patients. Resource utilization, including tissue and blood cultures, and use of radiographic analysis was also collected. The primary outcome was 96-h ED revisit/hospitalization. RESULTS: Analysis included 308 patients; 219 ED and 89 OU. OU patients were significantly more likely to be obese, have COPD/asthma, be diagnosed with cellulitis, and meet at least one systemic inflammatory response syndrome (SIRS) criterion. Tissue cultures were obtained in 21.7% of abscesses in the ED; 67.9% were in uncomplicated abscesses. In the OU tissue cultures were obtained in 48.8% of abscesses and 37.5% were uncomplicated cases. Blood cultures were drawn in 18.3% of ED patients and 56.2% of OU patients, not significantly associated with the presence of SIRS criteria. Radiology was used in the diagnosis of ABSSSIs in 33.5% of ED versus 69.5% OU patients (p < 0.001), Plain film radiograph being the most common. Thirty patients revisited the ED or required hospitalization within 96 h, 23 from the ED (p = 0.479). Prior history of ABSSSI (adjusted odds ratio [aOR] = 2.382, 95% confidence interval [CI] 1.264-6.346) and location on torso/buttocks (aOR = 2.355, 95% CI 1.067-5.197) were independent predictors. CONCLUSIONS: The low rate of ED revisit/hospitalization supports the use of OUs for low acuity ABSSSIs requiring initial IV therapy. Resource utilization within EDs/OUs for the management of ABSSSIs needs to be evaluated for unnecessary testing/procures.

Metacognitive therapy in treatment-resistant depression: a platform trial.

Patients with treatment-resistant depression received up to 8 sessions of metacognitive therapy (MCT) targeting attentional control, rumination, worry, and metacognitive beliefs. A baseline period was followed by weekly sessions with follow-up assessments at 6 and 12 months post treatment. Large and statistically significant improvements occurred in all symptom measures at post treatment and were maintained over follow-up. Two out of 3 process measures significantly improved at post treatment and all of these measures were improved at follow-up. Treatment was associated with similar response rates on the BDI and Hamilton rating scale. Using liberal criteria 80% of completers were classified as recovered at post treatment and 70% at follow-up on the BDI. In the intention to treat sample 66.6% were recovered at post treatment and 58.3% at follow-up. More stringent criteria showed 60% recovery rates at post treatment and at 12 m. The results suggest that MCT could be a brief and effective treatment and they provide a precedent for more definitive randomized controlled trials.

Auditory pseudohallucinations in United Kingdom war veterans and civilians with posttraumatic stress disorder.

OBJECTIVE: Hearing voices is a little-known feature of posttraumatic stress disorder (PTSD), mainly reported in US war veterans; it may be better conceived of as a dissociative than as a psychotic phenomenon. We investigated this feature in a pair of studies: Study 1 tested whether hearing voices was also reported by United Kingdom war veterans and whether it was associated with other dissociative reactions. Study 2 tested whether reports of hearing voices would generalize to a civilian sample, evaluated whether it was specific to PTSD or could be explained by trauma exposure alone, and investigated its phenomenological characteristics in more detail. METHOD: Study 1, which was conducted from 2005 to 2008 at numerous sites in the United Kingdom, contrasted male war veterans with current PTSD, past PTSD, and no PTSD on measures of dissociation and hearing voices. Study 2, which was conducted from 2004 to 2008 in London, United Kingdom, compared hearing voices in civilian patients with PTSD, healthy controls exposed to trauma, and depressed patients. RESULTS: Study 1 showed that more veterans with current or past PTSD than no PTSD described hearing voices, which was related to other dissociative reactions. Study 2 confirmed that hearing voices was also present in a civilian sample, that it was specific to PTSD, and that it had the characteristics of a pseudohallucination. CONCLUSIONS: The results emphasize the dissociative nature of PTSD, identify a little-known symptom that causes considerable distress, and suggest new directions for the assessment and treatment of PTSD in military and civilian populations.

Imagery rescripting as a brief stand-alone treatment for depressed patients with intrusive memories.

Many depressed patients report intrusive and distressing memories of specific events in their lives. Where present, these memories are believed to act as a maintaining factor. A series of ten patients with major depressive disorder and intrusive memories, many of them reporting severe, chronic, or recurrent episodes of depression, were given an average of 8.1 sessions of imagery rescripting as a stand-alone treatment. Hierarchical linear modelling demonstrated large treatment effects that were well maintained at one year follow-up. Seven patients showed reliable improvement, and six patients clinically significant improvement. These gains were achieved entirely by working through patients' visual imagination and without verbal challenging of negative beliefs. Spontaneous changes in beliefs, rumination, and behaviour were nevertheless observed.

I'll believe it when I can see it: imagery rescripting of intrusive sensory memories in depression.

Intrusive sensory memories are a common feature of depression but these are not targeted in standard cognitive treatments. Imagery rescripting of distressing memories has so far been mainly used to treat trauma-related disorders and as a component of the treatment of personality disorders. We propose that this approach might also be effective in treating depression. This paper describes the initial application of imagery rescripting as a stand-alone treatment for two patients with a sole diagnosis of major depressive disorder. The two cases are described in detail and follow-up data are reported. Implications for the cognitive treatment of depression and for our theoretical understanding about the mechanisms of change in cognitive therapy are considered.

Intrusive images and memories in major depression.

Individuals with current major depression were interviewed to investigate the prevalence of distressing intrusive mental imagery among depressed patients and study the phenomenology of these intrusions. Of the 39 currently depressed patients, 17 experienced some form of repetitive intrusive imagery (i.e., either an intrusive memory or image), with intrusive memories being more common than images. The intrusive imagery was experienced as highly uncontrollable and interfered significantly with patients' everyday lives. The intrusions were experienced with a sense of 'nowness', as well as physical and emotional re-experiencing. Despite high levels of re-experiencing, levels of dissociation were very low. The intrusive imagery was in some patients part of a wider network of key defining autobiographical memories, consistent with the idea that it is likely to play a significant role in maintaining the patient's depressive mood. Interventions targeting these intrusions could potentially result in a positive shift in depressed mood.

Autobiographical memory and social problem-solving in Asperger syndrome.

Difficulties in social interaction are a central feature of Asperger syndrome. Effective social interaction involves the ability to solve interpersonal problems as and when they occur. Here we examined social problem-solving in a group of adults with Asperger syndrome and control group matched for age, gender and IQ. We also assessed autobiographical memory, on a cueing task and during social problem-solving, and examined the relationship between access to specific past experiences and social problem-solving ability. Results demonstrated a social problem-solving impairment in the Asperger group. Their solutions were less detailed, less effective and less extended in time. Autobiographical memory performance was also impaired with significantly longer latencies to retrieve specific memories and fewer specific memories retrieved in comparison to controls.