[Effectivity of Inpatient and Day-clinic Psychosomatic-Psychotherapeutic Treatment]. / Effektivität stationärer und teilstationärer psychosomatisch-psychotherapeutischer Behandlung.

AIM: (Partial) inpatient psychotherapy is well implemented in Germany. To better understand efficacy factors and effects, efficacy studies are necessary. This naturalistic study investigates the effectiveness of inpatient and day clinic psychotherapy as well as patient-and disorder-related factors influencing individual symptom improvements. METHODS: Patients at a psychosomatic-psychotherapeutic university hospital treated from 2015 to 2019 who completed the BSI-18 either at admission and discharge (n=1366) or at admission and three-month catamnesis (n=497) were included in the analysis. RESULTS: Improvements in global symptom severity showed moderate effect sizes. Descriptively, these were larger for day clinic patients than for those receiving treatment as inpatients-especially in the follow-up comparison (immediately after discharge: dinpatient=0.401, dday clinic=0.482; three months after discharge: dinpatient=0.403, dday clinical=0.807). Day hospitalized patients differed significantly from inpatients-slightly in age, employment status, ability to work and initial symptom burden, moderately in the number of mental comorbidities and strongly in their main diagnoses. Socio-demographic factors showed no positive influence on symptom improvement, initial symptom severity a moderate positive influence and the number of mental comorbidities a complex influence. DISCUSSION: In general, this study confirms the effectiveness of (partial) inpatient psychosomatic therapy. The relevance of day clinic offers is emphasised in the context of cost efficiency and good integration into everyday life, but under consideration of individual treatment indications.

[Differential Reflection of the Relationship between Therapeutic Alliance and Outcome in a Day Treatment Hospital Setting]. / Differentielle Betrachtung des Zusammenhangs von therapeutischer Beziehung und Therapieerfolg in einem tagesklinischen Versorgungssetting.

OBJECTIVE: The therapeutic alliance is considered to be a significant and empirically well-documented determinant of therapeutical success. The aim of the present study was to replicate this effect using a large daily clinical sample and to consider different aspects of the therapeutic relationship in an extreme group of particularly low relationship satisfaction separately. METHOD: A longitudinal examination of a sample of n=809 patients (M=34,32; SD=10,7; 72,6% female) in a day care hospital setting was carried out. Using multiple regression analysis, the link between therapeutic alliance (Helping Alliance Questionnaire; HAQ-S) in the third week and therapeutic outcome (Brief Symptom Inventory-18; BSI-18) was investigated. Analyses have been conducted for the overall sample as well as for the extreme group representing the lowest decile regarding relationship satisfaction and the remaining 90%. A distinction was also made between the 2 subscales of the HAQ, satisfaction with the relationship and satisfaction with the outcome. RESULTS: The therapeutic alliance after 3 weeks was a significant predictor for therapeutic outcome. Within the extreme group with initially low relationship satisfaction, relationship satisfaction proved to be a significant and strong predictor for therapeutic outcome, but was not useable for the prediction of individual cases due to wide confidence interval (ß=0,622; 95% CI [0,051; 1,095]). In contrast, the relationship satisfaction in the remaining 90 percent explained no additional variance of therapeutic outcome beyond the explanation by satisfaction with the outcome (ß=0,244; 95% CI [0,176; 0,391]). DISCUSSION: The results emphasize the importance of therapeutic alliance also in a day care hospital setting. Relationship satisfaction plays a central, other factors exceeding role in the prediction of therapeutic outcome only in the extreme group of particularly dissatisfied patients. CONCLUSION: Ensuring at least a sufficiently good therapeutic relationship is of great importance and therefore requires early identification and, if necessary, intervention.

Transduction and transfection of difficult-to-transfect cells: Systematic attempts for the transfection of protozoa Leishmania.

Cell-penetrating peptides (CPPs) are used to internalize different cargoes, including DNA, into live mammalian and plant cells. Despite many cells being easily transfected with this approach, other cells are rather "difficult" or "hard to transfect," including protist cells of the genus Leishmania. Based on our previous results in successfully internalizing proteins into Leishmania tarentolae cells, we used single CPPs and three different DNA-binding proteins to form protein-like complexes with plasmids covered with CPPs. We attempted magnetofection, electroporation, and transfection using a number of commercially available detergents. While complex formation with negatively charged DNA required substantially higher amounts of CPPs than those necessary for mostly neutral proteins, the cytotoxicity of the required amounts of CPPs and auxiliaries was thoroughly studied. We found that Leishmania cells were indeed susceptible to high concentrations of some CPPs and auxiliaries, although in a different manner compared with that for mammalian cells. The lack of successful transfections implies the necessity to accept certain general limitations regarding DNA internalization into difficult-to-transfect cells. Only electroporation allowed reproducible internalization of large and rigid plasmid DNA molecules through electrically disturbed extended membrane areas, known as permeable membrane macrodomains.

Early maladaptive schemas in patients with somatoform disorders and somatization.

Maladaptive schemas are stable relational patterns that develop through harmful childhood experiences with primary caregivers. Schemas within somatoform disorders are rarely explored even though these disorders are clinically important due to high prevalence, co-morbidity, and cost for the health care system. This study investigates schemas according to Young's schema theory in patients with somatoform disorders in comparison with healthy controls and patients with depressive or anxiety disorders. Further associations between schemas and somatization were explored. We included 134 patients with a somatoform disorder and 39 age-matched healthy controls, 83 patients with a unipolar depression, and 34 patients with an anxiety disorder. The clinical sample consists of day care patients, diagnosed with the Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, but without a personality disorder. Primary measures were the Young Schema Questionnaire (YSQ-S2), the Screening for Somatoform Disorders (SOMS-7T), the Beck Depression Inventory, Second Edition, and the Childhood Trauma Questionnaire. Analyses of variance indicated that somatoform patients scored higher on almost all schemas than do healthy controls (p < 0.001, η2  = 0.148). The highest mean scores were reached for the schemas "self-sacrifice" and "unrelenting standards," with significant higher values in the patient sample. However, when compared with patients with depressive or anxiety disorders, somatoform patients scored equally or even lower. High somatization was associated with generally higher schema activation. This effect was to a great extent mediated by depressive symptoms. Only the schema "vulnerability to harm or illness" was exclusively related to somatization. These findings suggest that schemas should be systematically assessed within psychotherapy of somatoform patients.

Predicting attention deficit hyperactivity disorder using pregnancy and birth characteristics.

PURPOSE: The aim of this study was to evaluate maternal, prenatal, perinatal, and postpartum parameters as risk factors for the later development of an attention deficit hyperactivity disorder (ADHD) in the child. METHODS: Women who had given birth at Erlangen University Hospital between 1996 and 1999 were sent a questionnaire in 2009. The results of the questionnaire were correlated with the prospectively collected data for the births in 1996-1999. RESULTS: A total of 573 mother and child pairs were analyzed. Forty-four of the mothers reported that their child had ADHD (7.7%). No significant associations were found for the following parameters: mother's age; mother's educational level; number of the pregnancy; maternal weight before and at the end of pregnancy; mother's height; alcohol consumption during pregnancy; mode of delivery; gestational week; birthweight; umbilical artery blood values; Apgar score at 5 and 10 min; or breastfeeding. The parameters of smoking in pregnancy and an Apgar score lower than 7 after 1 min were significantly associated with a risk for later development of ADHD. CONCLUSIONS: This analysis of maternal, prenatal, perinatal, and postnatal parameters found that smoking in pregnancy and a low Apgar score 1 min after birth are associated with a significantly greater risk for the development of ADHD. Beyond the question of the causal mechanism involved, this is a relevant finding, since smoking during pregnancy is a preventable risk factor.

Quality Evaluation of Zirconium Dioxide Frameworks Produced in Five Dental Laboratories from Different Countries.

PURPOSE: The aim of this study was to assess and compare quality as well as economic aspects of CAD/CAM high strength ceramic three-unit FDP frameworks ordered from dental laboratories located in emerging countries and Switzerland. MATERIAL AND METHODS: The master casts of six cases were sent to five dental laboratories located in Thailand (Bangkok), China (Peking and Shenzhen), Turkey (Izmir), and Switzerland (Bern). Each laboratory was using a different CAD/CAM system. The clinical fit of the frameworks was qualitatively assessed, and the thickness of the framework material, the connector height, the width, and the diameter were evaluated using a measuring sensor. The analysis of the internal fit of the frameworks was performed by means of a replica technique, whereas the inner and outer surfaces of the frameworks were evaluated for traces of postprocessing and damage to the intaglio surface with light and electronic microscopes. Groups (dental laboratories and cases) were compared for statistically significant differences using Mann-Whitney U-tests after Bonferroni correction. RESULTS: An acceptable clinical fit was found at 97.9% of the margins produced in laboratory E, 87.5% in B, 93.7% in C, 79.2% in A, and 62.5% in D. The mean framework thicknesses were not statistically significantly different for the premolar regions; however, for the molar area 4/8 of the evaluated sites were statistically significantly different. Circumference, surface, and width of the connectors produced in the different laboratories were statistically significantly different but not the height. There were great differences in the designs for the pontic and connector regions, and some of the frameworks would not be recommended for clinical use. Traces of heavy postprocessing were found in frameworks from some of the laboratories. The prices per framework ranged from US$177 to US$896. CONCLUSIONS: By ordering laboratory work in developing countries, a considerable price reduction was obtained compared to the price level in Switzerland. Despite the use of the standardized CAD/CAM chains of production in all laboratories, a large variability in the quality aspects, such as clinical marginal fit, connector and pontic design, as well as postprocessing traces was noted. Recommended sound handling of postprocessing was not applied in all laboratories. Dentists should be aware of the true and factitious advantages of CAD/CAM production chains and not lose control over the process.

Optimization of the transfection of human THP-1 macrophages by application of Nunc UpCell technology.

We have established an electroporation protocol for transfection of premature adherent human THP-1 macrophages using Lonza Nucleofector technology. For efficient electroporation, detachment of adherent cells is necessary. We tested the Nunc UpCell product line of Thermo Fisher Scientific, which achieves detachment by a change of ambient temperature, as an alternative to enzymatic detachment. Here we present data verifying proper cell morphology and vitality and high transfection efficiency for macrophages cultured on UpCell plates. Appropriate macrophage behavior was confirmed by measuring markers of macrophage differentiation and polarization by reverse transcription quantitative polymerase chain reaction (RT-qPCR). In conclusion, Nunc UpCell materials are a viable alternative to enzymatic detachment.

Vessel transformation in chronic wounds under topical negative pressure therapy: an immunohistochemical analysis.

The underlying physiological mechanism of topical negative pressure (TNP) therapy is not yet completely understood. This prospective clinical study aims to clarify a potential influence of TNP therapy on vessel proliferation and hypoxia in chronic wounds. TNP was applied on chronic wounds of 16 patients (-125 mmHg) to prepare them for a plastic-surgical reconstruction using free or pedicled flaps. Tissue biopsies were taken from the wound edge and wound bed at different time points. All samples were stained with haematoxylin and eosin, hypoxia-induced factor-1α and endothelial cell markers (CD31 and CD34) for the immunohistological analysis of inflammation, hypoxia and vessel proliferation. Between day 5 and day 8 of treatment, a considerable increase in blood vessel density could be observed, reaching a maximum of approximately 200% in contrast to the vessel density prior to treatment. In addition, the number of hypoxic and inflammatory cells was found to be increased at particular time points. This study demonstrates a stimulating effect on vessel proliferation under TNP treatment. TNP appears to support (neo-) angiogenesis and transformation of chronic non-healing wounds in a physiological wound healing process when combined with surgical debridement. This effect underlines the positive influence of TNP in the treatment of chronic wounds as shown by various clinical reports.

Transduction of proteins into leishmania tarentolae by formation of non-covalent complexes with cell-penetrating peptides.

Cell-penetrating peptides (CPPs) are used to transport peptides, proteins, different types of ribonucleic acids (or mimics of these molecules), and DNA into live cells, both plant and mammalian. Leishmania belongs to the class of protozoa having, in comparison to mammalian cells, a different lipid composition of the membrane, proteoglycans on the surface, and signal pathways. We investigated the uptake of two different and easily detectable proteins into the non-pathogenic strain Leishmania tarentolae. From the large number of CPPs available, six and a histone were chosen specifically for their ability to form non-covalent complexes. For Leishmania we used the enzyme ß-galactosidase and fluorescent labeled bovine serum albumin as cargoes. The results are compared to similar internalization studies using mammalian cells [Mussbach et al., ]. Leishmania cells can degrade CPPs by a secreted and membrane-bound chymotrypsin-like protease. Both cargo proteins were internalized with sufficient efficiency and achieved intramolecular concentrations similar to mammalian cells. The transport efficiencies of the CPPs differed from each other, and showed a different rank order for both cargoes. The intracellular distribution of fluorescent-labeled bovine serum albumin showed highest concentrations in the nucleus and kinetoplast. Leishmania are susceptible to high concentrations of some CPPs, although comparably dissimilar to mammalian cells. MPG-peptides are more cytotoxic in Leishmania than in mammalian cells, acting as antimicrobial peptides. Our results contribute to a better understanding of molecular interactions in Leishmania cells and possibly to new treatments of leishmaniasis.

Complexity of fatty acid distribution inside human macrophages on single cell level using Raman micro-spectroscopy.

Macrophages are phagocytic cells which are involved in the non-specific immune defense. Lipid uptake and storage behavior of macrophages also play a key role in the development of atherosclerotic lesions within walls of blood vessels. The allocation of exogenous lipids such as fatty acids in the blood stream dictates the accumulation and quantity of lipids within macrophages. In case of an overexposure, macrophages transform into foam cells because of the large amount of lipid droplets in the cytoplasm. Raman micro-spectroscopy is a powerful tool for studying single cells due to the combination of microscopic imaging with spectral information. With a spatial resolution restricted by the diffraction limit, it is possible to visualize lipid droplets within macrophages. With stable isotopic labeling of fatty acids with deuterium, the uptake and storage of exogenously provided fatty acids can be investigated. In this study, we present the results of time-dependent Raman spectroscopic imaging of single THP-1 macrophages incubated with deuterated arachidonic acid. The polyunsaturated fatty acid plays an important role in the cellular signaling pathway as being the precursor of icosanoids. We show that arachidonic acid is stored in lipid droplets but foam cell formation is less pronounced as with other fatty acids. The storage efficiency in lipid droplets is lower than in cells incubated with deuterated palmitic acid. We validate our results with gas chromatography and gain information on the relative content of arachidonic acid and its metabolites in treated macrophages. These analyses also provide evidence that significant amounts of the intracellular arachidonic acid is elongated to adrenic acid but is not metabolized any further. The co-supplementation of deuterated arachidonic acid and deuterated palmitic acid leads to a non-homogenous storage pattern in lipid droplets within single cells.

The Third Annual Symposium of the Midwest Aging Consortium.

The geroscience hypothesis suggests that addressing the fundamental mechanisms driving aging biology will prevent or mitigate the onset of multiple chronic diseases, for which the largest risk factor is advanced age. Research that investigates the root causes of aging is therefore of critical importance given the rising healthcare burden attributable to age-related diseases. The third annual Midwest Aging Consortium symposium was convened as a showcase of such research performed by investigators from institutions across the Midwestern United States. This report summarizes the work presented during a virtual conference across topics in aging biology, including immune function in the lung-particularly timely given the Corona Virus Immune Disease-2019 pandemic-along with the role of metabolism and nutrient-regulated pathways in cellular function with age, the influence of senescence on stem cell function and inflammation, and our evolving understanding of the mechanisms underlying observation of sex dimorphism in aging-related outcomes. The symposium focused on early-stage and emerging investigators, while including keynote presentations from leaders in the biology of aging field, highlighting the diversity and strength of aging research in the Midwest.

The alliance-outcome association in borderline and obsessive-compulsive personality disorder.

Personality disorders are considered a possible factor affecting the relationship between therapeutic alliance and therapy outcome. The present study investigated the alliance-outcome effect in patient groups with borderline personality disorder (BPD) and obsessive-compulsive personality disorder (OCPD). Data derived from a sample of n = 66 patients, treated in a day care hospital setting with a dialectical-behavioral and schema therapeutic treatment concept. Patients rated their symptom severity at admission, early alliance after 4-6 therapy sessions and symptom severity as well as alliance at discharge. Results showed no significant differences between BPD and OCPD patients regarding symptom severity and alliance. Multiple regression analyses indicated that the alliance was a significant predictor of symptom reduction, however only in the OCPD group. Our results showed an exceptionally strong alliance-outcome relationship in OCPD patients, suggesting that focusing on building a strong alliance and measuring it early in therapy may be especially beneficial for this patient group. For patients with BPD, a more regular screening of the therapeutic alliance might be helpful.

Subtypes of borderline personality disorder in a day-clinic setting-Clinical and therapeutic differences.

This preliminary study aims at extending existing empirical evidence on subtypes of borderline personality disorders (BPDs) by identifying subtypes among patients with BPD, comparing their characteristics to the trait domains of the dimensional model of the International Classification of Diseases, 11th Revision (ICD-11; World Health Organization [WHO], 2022), and examining differences in sociodemographic, clinical, and therapeutic outcome variables. Data of N = 109 patients were subjected to cluster analysis based on the International Personality Disorder Examination variables for BPD and analyzed regarding differences in clinical and therapeutic variables. Clustering suggested a three-cluster solution, namely, internalizing (n = 35), externalizing (n = 28), and mixed subtype (n = 46). Subtypes showed differences in clinical variables and therapeutic outcomes with the internalizing showing more affective disorders and the mixed subtype showing the lowest therapeutic change in borderline-specific symptoms. Together, the present results correspond to the model of the ICD-11 (WHO, 2022). Clinical and treatment implications are being discussed. (PsycInfo Database Record (c) 2023 APA, all rights reserved).

Population pharmacokinetics of riociguat in a pediatric population (aged ≥ 6 years) with pulmonary arterial hypertension.

BACKGROUND: The PATENT-CHILD study investigated riociguat in children aged ≥ 6 to <18 years with pulmonary arterial hypertension (PAH) treated with tablets or an oral pediatric suspension based on bodyweight-adjusted dosing of up to 2.5 mg three times daily. PATENT-CHILD demonstrated an acceptable riociguat safety profile and individual plasma concentrations in pediatric patients were consistent with those in adult patients. METHODS: Using the data set from PATENT-CHILD and building on existing population pharmacokinetic (PK) models for riociguat and its major metabolite (M1) in adults with PAH, a coupled riociguat-M1 PK model was developed. The final model developed incorporated a one-compartment model for riociguat, coupled to a one-compartment model for M1, allowing for presystemic formation of M1. It included allometric scaling exponents for bodyweight. RESULTS: Apparent clearance of riociguat was similar in children and adult patients with PAH (median [interquartile range] 2.20 [1.75-3.44] and 2.08 L/h [1.55-2.97]). Factors contributing to lower PK exposure were lower riociguat maintenance dose in PATENT-CHILD, and a higher riociguat clearance in some adolescent patients, compared with adult patients. No effects of formulation, sex, or age on riociguat PK were observed. An exploratory PK/pharmacodynamics analysis found the increase in 6-min walking distance in pediatric patients treated with riociguat was not related to riociguat PK. CONCLUSIONS: Body size is the main determinant of PK in growing children, and the model supports clinical data that, for children weighing < 50 kg, a bodyweight-adjusted dose of riociguat should be used to achieve a similar exposure to that observed in adults with PAH.

Immunometabolism at the crossroads of obesity and cancer-a Keystone Symposia report.

Immunometabolism considers the relationship between metabolism and immunity. Typically, researchers focus on either the metabolic pathways within immune cells that affect their function or the impact of immune cells on systemic metabolism. A more holistic approach that considers both these viewpoints is needed. On September 5-8, 2022, experts in the field of immunometabolism met for the Keystone symposium "Immunometabolism at the Crossroads of Obesity and Cancer" to present recent research across the field of immunometabolism, with the setting of obesity and cancer as an ideal example of the complex interplay between metabolism, immunity, and cancer. Speakers highlighted new insights on the metabolic links between tumor cells and immune cells, with a focus on leveraging unique metabolic vulnerabilities of different cell types in the tumor microenvironment as therapeutic targets and demonstrated the effects of diet, the microbiome, and obesity on immune system function and cancer pathogenesis and therapy. Finally, speakers presented new technologies to interrogate the immune system and uncover novel metabolic pathways important for immunity.

Radiation dose considerations by intra-individual Monte Carlo simulations in dual source spiral coronary computed tomography angiography with electrocardiogram-triggered tube current modulation and adaptive pitch.

OBJECTIVES: To evaluate radiation dose levels in patients undergoing spiral coronary computed tomography angiography (CTA) on a dual-source system in clinical routine. METHODS: Coronary CTA was performed for 56 patients with electrocardiogram-triggered tube current modulation (TCM) and heart-rate (HR) dependent pitch adaptation. Individual Monte Carlo (MC) simulations were performed for dose assessment. Retrospective simulations with constant tube current (CTC) served as reference. Lung tissue was segmented and used for organ and effective dose (ED) calculation. RESULTS: Estimates for mean relative ED was 7.1 ± 2.1 mSv/100 mAs for TCM and 12.5 ± 5.3 mSv/100 mAs for CTC (P < 0.001). Relative dose reduction at low HR (≤60 bpm) was highest (49 ± 5%) compared to intermediate (60-70 bpm, 33 ± 12%) and high HR (>70 bpm, 29 ± 12%). However lowest ED is achieved at high HR (5.2 ± 1.5 mSv/100 mAs), compared with intermediate (6.7 ± 1.6 mSv/100 mAs) and low (8.3 ± 2.1 mSv/100 mAs) HR when automated pitch adaptation is applied. CONCLUSIONS: Radiation dose savings up to 52% are achievable by TCM at low and regular HR. However lowest ED is attained at high HR by pitch adaptation despite inferior radiation dose reduction by TCM. KEY POINTS: • Monte Carlo simulations allow for individual radiation dose calculations. • ECG-triggered tube current modulation (TCM) can effectively reduce radiation dose. • Slow and regular heart rates allow for highest dose reductions by TCM. • Adaptive pitch accounts for lowest radiation dose at high heart rates. • Women receive higher effective dose than men undergoing spiral coronary CT-angiography.

Metabolic Regulation of Stem Cells in Aging.

PURPOSE OF REVIEW: From invertebrates to vertebrates, the ability to sense nutrient availability is critical for survival. Complex organisms have evolved numerous signaling pathways to sense nutrients and dietary fluctuations, which influence many cellular processes. Although both overabundance and extreme depletion of nutrients can lead to deleterious effects, dietary restriction without malnutrition can increase lifespan and promote overall health in many model organisms. In this review, we focus on age-dependent changes in stem cell metabolism and dietary interventions used to modulate stem cell function in aging. RECENT FINDINGS: Over the last half-century, seminal studies have illustrated that dietary restriction confers beneficial effects on longevity in many model organisms. Many researchers have now turned to dissecting the molecular mechanisms by which these diets affect aging at the cellular level. One subpopulation of cells of particular interest are adult stem cells, the most regenerative cells of the body. It is generally accepted that the regenerative capacity of stem cells declines with age, and while the metabolic requirements of each vary across tissues, the ability of dietary interventions to influence stem cell function is striking. SUMMARY: In this review, we will focus primarily on how metabolism plays a role in adult stem cell homeostasis with respect to aging, with particular emphasis on intestinal stem cells while also touching on hematopoietic, skeletal muscle, and neural stem cells. We will also discuss key metabolic signaling pathways influenced by both dietary restriction and the aging process, and will examine their role in improving tissue homeostasis and lifespan. Understanding the mechanisms behind the metabolic needs of stem cells will help bridge the divide between a basic science interpretation of stem cell function and a whole-organism view of nutrition, thereby providing insight into potential dietary or therapeutic interventions.

Inhibition of Bromodomain and Extra Terminal (BET) Domain Activity Modulates the IL-23R/IL-17 Axis and Suppresses Acute Graft-Versus-Host Disease.

Acute graft-versus-host disease (GVHD) is the leading cause of non-relapse mortality following allogeneic hematopoietic cell transplantation. The majority of patients non-responsive to front line treatment with steroids have an estimated overall 2-year survival rate of only 10%. Bromodomain and extra-terminal domain (BET) proteins influence inflammatory gene transcription, and therefore represent a potential target to mitigate inflammation central to acute GVHD pathogenesis. Using potent and selective BET inhibitors Plexxikon-51107 and -2853 (PLX51107 and PLX2853), we show that BET inhibition significantly improves survival and reduces disease progression in murine models of acute GVHD without sacrificing the beneficial graft-versus-leukemia response. BET inhibition reduces T cell alloreactive proliferation, decreases inflammatory cytokine production, and impairs dendritic cell maturation both in vitro and in vivo. RNA sequencing studies in human T cells revealed that BET inhibition impacts inflammatory IL-17 and IL-12 gene expression signatures, and Chromatin Immunoprecipitation (ChIP)-sequencing revealed that BRD4 binds directly to the IL-23R gene locus. BET inhibition results in decreased IL-23R expression and function as demonstrated by decreased phosphorylation of STAT3 in response to IL-23 stimulation in human T cells in vitro as well as in mouse donor T cells in vivo. Furthermore, PLX2853 significantly reduced IL-23R+ and pathogenic CD4+ IFNγ+ IL-17+ double positive T cell infiltration in gastrointestinal tissues in an acute GVHD murine model. Our findings identify a role for BET proteins in regulating the IL-23R/STAT3/IL-17 pathway. Based on our preclinical data presented here, PLX51107 will enter clinical trial for refractory acute GVHD in a Phase 1 safety, biological efficacy trial.

Fibrinonecrotic Placentitis and Abortion Associated With Pantoea agglomerans Infection in a Mare.

Pantoea agglomerans, family Enterobacteriaceae, is a Gram-negative bacterium that may be isolated from soil and from plants. This bacterium has been associated with disease in plants, humans, and rarely in domestic animal species. We describe here a case of fibrinonecrotic placentitis and equine abortion associated with P. agglomerans infection in southern Brazil. A fetus with 10 months of gestation and its placenta were evaluated. Gross lesions were observed in the cervical star extending to the body of the chorioallantois and consisted of a focally extensive, transmural, severely thickened yellow area. Histologically, this area in the chorioallantois was effaced by severe necrosis, associated with marked inflammatory infiltrate of neutrophils and abundant deposition of fibrin and cellular debris. Aggregates of bacterial rods were noted intermixed with inflammation areas. No significant lesions were observed in the remaining organs inspected. Tissue samples of the lung, placenta, and stomach contents were cultured, and microbiological tests revealed the growth of P. agglomerans in all evaluated samples. The present study reaffirms the participation of P. agglomerans as a cause of bacterial placentitis and abortion in horses.

Transfecting Macrophages.

Transfection is defined as the transfer of foreign nucleic acids into cells. In general, transfection may achieve either overexpression of a gene by the transfer of plasmid DNA or suppression of gene expression by RNA interference after transfer of small interfering RNA. Both approaches allow for the detailed investigation of the function of a particular gene product or mechanisms of gene regulation. Macrophages are considered as hard-to-transfect cells, as they have evolved to recognize foreign nucleic acids and to initiate an immune response to these molecules. The presented electroporation protocol provides an effective tool to efficiently transfect human THP-1 macrophages with siRNA or plasmid DNA while avoiding macrophage activation.