Control of motivation for sucrose in the paraventricular hypothalamic nucleus by dynorphin peptides and the kappa opioid receptor.

The dynorphin peptides are the endogenous ligands for the kappa opioid receptor (KOR) and regulate food intake. Administration of dynorphin-A1-13 (DYN) in the paraventricular hypothalamic nucleus (PVN) increases palatable food intake, and this effect is blocked by co-administration of the orexin-A neuropeptide, which is co-released with DYN in PVN from neurons located in the lateral hypothalamus. While PVN administration of DYN increases palatable food intake, whether it increases food-seeking behaviors has yet to be examined. We tested the effects of DYN and norBNI (a KOR antagonist) on the seeking and consumption of sucrose using a progressive ratio (PR) and demand curve (DC) tasks. In PVN, DYN did not alter the sucrose breaking point (BP) in the PR task nor the elasticity or intensity of demand for sucrose in the DC task. Still, DYN reduced the delay in obtaining sucrose and increased licks during sucrose intake in the PR task, irrespective of the co-administration of orexin-A. In PVN, norBNI increased the delay in obtaining sucrose and reduced licks during sucrose intake in the PR task while increasing elasticity without altering intensity of demand in the DC task. However, subcutaneous norBNI reduced the BP for sucrose and increased the delay in obtaining sucrose in the PR task while reducing the elasticity of demand. Together, these data show different effects of systemic and PVN blockade of KOR on food-seeking, consummatory behaviors, and incentive motivation for sucrose and suggest that KOR activity in PVN is necessary but not sufficient to drive seeking behaviors for palatable food.

Pax3 induces target-specific reinnervation through axon collateral expression of PSA-NCAM.

Damaged or dysfunctional neural circuits can be replaced after a lesion by axon sprouting and collateral growth from undamaged neurons. Unfortunately, these new connections are often disorganized and rarely produce clinical improvement. Here we investigate how to promote post-lesion axonal collateral growth, while retaining correct cellular targeting. In the mouse olivocerebellar path, brain-derived neurotrophic factor (BDNF) induces correctly-targeted post-lesion cerebellar reinnervation by remaining intact inferior olivary axons (climbing fibers). In this study we identified cellular processes through which BDNF induces this repair. BDNF injection into the denervated cerebellum upregulates the transcription factor Pax3 in inferior olivary neurons and induces rapid climbing fiber sprouting. Pax3 in turn increases polysialic acid-neural cell adhesion molecule (PSA-NCAM) in the sprouting climbing fiber path, facilitating collateral outgrowth and pathfinding to reinnervate the correct targets, cerebellar Purkinje cells. BDNF-induced reinnervation can be reproduced by olivary Pax3 overexpression, and abolished by olivary Pax3 knockdown, suggesting that Pax3 promotes axon growth and guidance through upregulating PSA-NCAM, probably on the axon's growth cone. These data indicate that restricting growth-promotion to potential reinnervating afferent neurons, as opposed to stimulating the whole circuit or the injury site, allows axon growth and appropriate guidance, thus accurately rebuilding a neural circuit.

Interobserver reliability of classifying shoulder calcific tendinopathy on plain radiography and ultrasound.

INTRODUCTION: Shoulder calcific tendinopathy is a frequent cause of shoulder pain. Diagnosis is usually based on ultrasound (US) and/or X-ray. US is considered an inherently operator-dependent imaging modality and, interobserver variability has previously been described by experts in the musculoskeletal US. The main objective of this study is to assess the interobserver agreement for shoulder calcific tendinopathy attending to the size, type, and location of calcium analyzed in plain film and ultrasound among trained musculoskeletal radiologists. MATERIAL AND METHODS: From June 2018 to May 2019, we conducted a prospective study. Patients diagnosed with shoulder pain related to calcific tendinopathy were included. Two different experienced musculoskeletal radiologists evaluated independently the plain film and the US. RESULTS: Forty patients, with a mean age of 54.6 years, were included. Cohen's kappa coefficient of 0.721 and 0.761 was obtained for the type of calcium encountered in plain film and the US, respectively. The location of calcification obtained a coefficient of 0.927 and 0.760 in plain film and US, respectively. The size of the calcification presented an intraclass correlation coefficient (ICC) of 0.891 and 0.86 in plain film and US respectively. No statistically significant differences were found in either measurement. CONCLUSION: This study shows very good interobserver reliability of type and size measurement (plain film and US) of shoulder calcifying tendinopathy in experienced musculoskeletal radiologists.

Meta-analysis of pre-clinical studies on the effects of opioid receptor ligands on food intake, motivation, and choice.

The opioid receptors (OR) regulate food intake. Still, despite extensive pre-clinical research, the overall effects and individual contribution of the mu (MOR), kappa (KOR), and delta (DOR) OR subtypes to feeding behaviors and food intake remain unclear. To address this, we conducted a pre-registered systematic search and meta-analysis of rodent dose-response studies to evaluate the impact of central and peripheral administration of non-selective and selective OR ligands on intake, motivation, and choice of food. All studies had a high bias risk. Still, the meta-analysis confirmed the overall orexigenic and anorexigenic effects of OR agonists and antagonists, respectively. Our results support a larger orexigenic role for central MOR agonists among OR subtypes and that peripheral OR antagonists reduce motivation for and intake of preferred foods. In binary food choice studies, peripheral OR agonists selectively increase the intake of fat-preferred foods; in contrast, they did not increase the intake of sweet carbohydrate-preferred foods. Overall, these data support that OR regulation of intake, motivation, and choice is influenced by food macronutrient composition.

EBAG9 silencing exerts an immune checkpoint function without aggravating adverse effects.

Chimeric antigen receptor (CAR) T cells have revolutionized treatment of B cell malignancies. However, enhancing the efficacy of engineered T cells without compromising their safety is warranted. The estrogen receptor-binding fragment-associated antigen 9 (EBAG9) inhibits release of cytolytic enzymes from cytotoxic T lymphocytes. Here, we examined the potency of EBAG9 silencing for the improvement of adoptive T cell therapy. MicroRNA (miRNA)-mediated EBAG9 downregulation in transplanted cytolytic CD8+ T cells (CTLs) from immunized mice improved their cytolytic competence in a tumor model. In tolerant female recipient mice that received organ transplants, a minor histocompatibility antigen was turned into a rejection antigen by Ebag9 deletion, indicating an immune checkpoint function for EBAG9. Considerably fewer EBAG9-silenced human CAR T cells were needed for tumor growth control in a xenotransplantation model. Transcriptome profiling did not reveal additional risks regarding genotoxicity or aberrant differentiation. A single-step retrovirus transduction process links CAR or TCR expression with miRNA-mediated EBAG9 downregulation. Despite higher cytolytic efficacy, release of cytokines associated with cytokine release syndrome remains unaffected. Collectively, EBAG9 silencing enhances effector capacity of TCR- and CAR-engineered T cells, results in improved tumor eradication, facilitates efficient manufacturing, and decreases the therapeutic dose.

Eficacia clínica de los umbrales de intervención híbridos y dependientes de la edad basados en FRAX en la población ecuatoriana / Clinical efficacy of FRAX-based age-dependent and hybrid intervention thresholds in the Ecuadorian population

Objetivo: Evaluar el impacto clínico de los umbrales de intervención basados en FRAX en mujeres ecuatorianas. Probar una combinación de umbrales de intervención fijo y específico de la edad para optimizar la selección de mujeres elegibles para intervención.Pacientes y métodos: Estudio transversal. Se seleccionaron 2.283 mujeres de 60 a 94 años. Calculamos el riesgo de fracturas osteoporóticas principales y de cuello de fémur con el modelo FRAX ecuatoriano (versión 4.1), y calculamos la proporción de individuos elegibles para tratamiento y evaluación de la densidad mineral ósea aplicando los umbrales específicos de edad de 60 a 94 años y un umbral fijo a partir de 75 años.Resultados: Aplicando los umbrales específicos de la edad, 2% de las mujeres calificaron para tratamiento y 73,7% para evaluación de la densidad mineral ósea. Según la edad, las mujeres elegibles para tratamiento fluctuaron entre 0,7 a 3,8% y las elegibles para evaluación de la densidad mineral ósea entre 58,3 al 80,5%.Con el umbral fijo, 31% de las mujeres calificaron para tratamiento y 76,3% para evaluación de la densidad mineral ósea. Dependiendo de la edad, las mujeres potencialmente elegibles para tratamiento fluctuaron de 3,8 a 76,5%, y las elegibles para evaluación de la densidad mineral ósea entre 65,2 al 85,4%.Conclusiones: La proporción de mujeres potencialmente elegible para el tratamiento, es baja comparada con países con riesgo alto de fracturas. Utilizar un umbral fijo a partir de los 75 años, optimiza la proporción de mujeres elegibles para tratamiento. En los países con riesgo de fractura bajo a moderado, con recursos limitados, un modelo híbrido puede ser más adecuado. (AU)

CXCR4 mediates leukemic cell migration and survival in the testicular microenvironment.

The testis is the second most frequent extramedullary site of relapse in pediatric acute lymphoblastic leukemia (ALL). The mechanism for B-cell (B) ALL cell migration towards and survival within the testis remains elusive. Here, we identified CXCL12-CXCR4 as the leading signaling axis for B-ALL cell migration and survival in the testicular leukemic niche. We combined analysis of primary human ALL with a novel patient-derived xenograft (PDX)-ALL mouse model with testicular involvement. Prerequisites for leukemic cell infiltration in the testis were prepubertal age of the recipient mice, high surface expression of CXCR4 on PDX-ALL cells, and CXCL12 secretion from the testicular stroma. Analysis of primary pediatric patient samples revealed that CXCR4 was the only chemokine receptor being robustly expressed on B-ALL cells both at the time of diagnosis and relapse. In affected patient testes, leukemic cells localized within the interstitial space in close proximity to testicular macrophages. Mouse macrophages isolated from affected testes, in the PDX model, revealed a macrophage polarization towards a M2-like phenotype in the presence of ALL cells. Therapeutically, blockade of CXCR4-mediated functions using an anti-CXCR4 antibody treatment completely abolished testicular infiltration of PDX-ALL cells and strongly impaired the overall development of leukemia. Collectively, we identified a prepubertal condition together with high CXCR4 expression as factors affecting the leukemia permissive testicular microenvironment. We propose CXCR4 as a promising target for therapeutic prevention of testicular relapses in childhood B-ALL. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

Accelerating clinical-scale production of BCMA CAR T cells with defined maturation stages.

The advent of CAR T cells targeting CD19 or BCMA on B cell neoplasm demonstrated remarkable efficacy, but rapid relapses and primary refractoriness remains challenging. A leading cause of CAR T cell failure is their lack of expansion and limited persistence. Long-lived, self-renewing multipotent T memory stem cells (TSCM) and T central memory cells (TCM) likely sustain superior tumor regression, but their low frequencies in blood from cancer patients impose a major hurdle for clinical CAR T production. We designed a clinically compliant protocol for generating BCMA CAR T cells starting with increased TSCM/TCM cell input. A CliniMACS Prodigy process was combined with flow cytometry-based enrichment of CD62L+CD95+ T cells. Although starting with only 15% of standard T cell input, the selected TSCM/TCM material was efficiently activated and transduced with a BCMA CAR-encoding retrovirus. Cultivation in the presence of IL-7/IL-15 enabled the harvest of CAR T cells containing an increased CD4+ TSCM fraction and 70% TSCM cells amongst CD8+. Strong cell proliferation yielded cell numbers sufficient for clinical application, while effector functions were maintained. Together, adaptation of a standard CliniMACS Prodigy protocol to low input numbers resulted in efficient retroviral transduction with a high CAR T cell yield.

Comparison of FACS and PCR for Detection of BCMA-CAR-T Cells.

Chimeric-antigen-receptor (CAR)-T-cell therapy is already widely used to treat patients who are relapsed or refractory to chemotherapy, antibodies, or stem-cell transplantation. Multiple myeloma still constitutes an incurable disease. CAR-T-cell therapy that targets BCMA (B-cell maturation antigen) is currently revolutionizing the treatment of those patients. To monitor and improve treatment outcomes, methods to detect CAR-T cells in human peripheral blood are highly desirable. In this study, three different detection reagents for staining BCMA-CAR-T cells by flow cytometry were compared. Moreover, a quantitative polymerase chain reaction (qPCR) to detect BCMA-CAR-T cells was established. By applying a cell-titration experiment of BCMA-CAR-T cells, both methods were compared head-to-head. In flow-cytometric analysis, the detection reagents used in this study could all detect BCMA-CAR-T cells at a similar level. The results of false-positive background staining differed as follows (standard deviation): the BCMA-detection reagent used on the control revealed a background staining of 0.04% (±0.02%), for the PE-labeled human BCMA peptide it was 0.25% (±0.06%) and for the polyclonal anti-human IgG antibody it was 7.2% (±9.2%). The ability to detect BCMA-CAR-T cells down to a concentration of 0.4% was similar for qPCR and flow cytometry. The qPCR could detect even lower concentrations (0.02-0.01%). In summary, BCMA-CAR-T-cell monitoring can be reliably performed by both flow cytometry and qPCR. In flow cytometry, reagents with low background staining should be preferred.

Using complexity-entropy planes to detect Parkinson's disease from short segments of haemodynamic signals.

Objective. There is emerging evidence that analysing the entropy and complexity of biomedical signals can detect underlying changes in physiology which may be reflective of disease pathology. This approach can be used even when only short recordings of biomedical signals are available. This study aimed to determine whether entropy and complexity measures can detect differences between subjects with Parkinsons disease and healthy controls (HCs).Approach. A method based on a diagram of entropy versus complexity, named complexity-entropy plane, was used to re-analyse a dataset of cerebral haemodynamic signals from subjects with Parkinsons disease and HCs obtained under poikilocapnic conditions. A probability distribution for a set of ordinal patterns, designed to capture regularities in a time series, was computed from each signal under analysis. Four types of entropy and ten types of complexity measures were estimated from these distributions. Mean values of entropy and complexity were compared and their classification power was assessed by evaluating the best linear separator on the corresponding complexity-entropy planes.Main results. Few linear separators obtained significantly better classification, evaluated as the area under the receiver operating characteristic curve, than signal mean values. However, significant differences in both entropy and complexity were detected between the groups of participants.Significance. Measures of entropy and complexity were able to detect differences between healthy volunteers and subjects with Parkinson's disease, in poikilocapnic conditions, even though only short recordings were available for analysis. Further work is needed to refine this promising approach, and to help understand the findings in the context of specific pathophysiological changes.

CXCR5 CAR-T cells simultaneously target B cell non-Hodgkin's lymphoma and tumor-supportive follicular T helper cells.

CAR-T cell therapy targeting CD19 demonstrated strong activity against advanced B cell leukemia, however shows less efficacy against lymphoma with nodal dissemination. To target both B cell Non-Hodgkin's lymphoma (B-NHLs) and follicular T helper (Tfh) cells in the tumor microenvironment (TME), we apply here a chimeric antigen receptor (CAR) that recognizes human CXCR5 with high avidity. CXCR5, physiologically expressed on mature B and Tfh cells, is also highly expressed on nodal B-NHLs. Anti-CXCR5 CAR-T cells eradicate B-NHL cells and lymphoma-supportive Tfh cells more potently than CD19 CAR-T cells in vitro, and they efficiently inhibit lymphoma growth in a murine xenograft model. Administration of anti-murine CXCR5 CAR-T cells in syngeneic mice specifically depletes endogenous and malignant B and Tfh cells without unexpected on-target/off-tumor effects. Collectively, anti-CXCR5 CAR-T cells provide a promising treatment strategy for nodal B-NHLs through the simultaneous elimination of lymphoma B cells and Tfh cells of the tumor-supporting TME.

Susceptibility to organophosphates pesticides and the development of infectious-contagious respiratory diseases.

In this paper we develop an SIRS compartmental model to investigate the dynamic interplay between pesticide intoxication and the spread of infectious-contagious respiratory diseases. We are particularly interested in investigating three levels of genetic susceptibility to pesticide intoxication. The genotypic distribution of susceptibility to pesticide intoxication, is proposed and parameterized according to ethnic variation using real population data from published studies, and we assume that pesticide intoxication increases susceptibility to infection with a respiratory pathogen. We use mathematical models to illustrate the impact of this distribution on the spread of hypothetical respiratory disease in a population exposed to the organophosphate pesticide. In this context, we show how an initial basic reproductive number below the epidemic threshold of 1.0 could be enhanced to support epidemic outbreaks in agricultural populations that employ chlorpyrifos pesticides. We further illustrate our modeling framework to study the effect of ethnic group variation in Singapore (Malay, Indian and Chinese) using genetic distribution data from published studies.

Dynamics between infectious diseases with two susceptibility conditions: A mathematical model.

This paper presents a novel epidemiological transmission model of a population affected by two different susceptible-infected-susceptible infectious diseases. For each disease, individuals fall into one of the two susceptibility conditions in which one of the diseases has the highest occurrence level. This model is unique in assuming that: (a) if an individual is infected by one disease, their susceptibility to the other disease is increased; (b) when an individual recovers from a disease they become less susceptible to it, i.e. they acquire partial immunity. The model captures these two assumptions by utilizing a coupled system of differential equations. Dynamic analysis of the system is based on basic reproductive number theory, and pattern visualization was performed using numerical simulation.

Hepatic JAK2 protects against atherosclerosis through circulating IGF-1.

Atherosclerosis is considered both a metabolic and inflammatory disease; however, the specific tissue and signaling molecules that instigate and propagate this disease remain unclear. The liver is a central site of inflammation and lipid metabolism that is critical for atherosclerosis, and JAK2 is a key mediator of inflammation and, more recently, of hepatic lipid metabolism. However, precise effects of hepatic Jak2 on atherosclerosis remain unknown. We show here that hepatic Jak2 deficiency in atherosclerosis-prone mouse models exhibited accelerated atherosclerosis with increased plaque macrophages and decreased plaque smooth muscle cell content. JAK2's essential role in growth hormone signalling in liver that resulted in reduced IGF-1 with hepatic Jak2 deficiency played a causal role in exacerbating atherosclerosis. As such, restoring IGF-1 either pharmacologically or genetically attenuated atherosclerotic burden. Together, our data show hepatic Jak2 to play a protective role in atherogenesis through actions mediated by circulating IGF-1 and, to our knowledge, provide a novel liver-centric mechanism in atheroprotection.

FAK signalling controls insulin sensitivity through regulation of adipocyte survival.

Focal adhesion kinase (FAK) plays a central role in integrin signalling, which regulates growth and survival of tumours. Here we show that FAK protein levels are increased in adipose tissue of insulin-resistant obese mice and humans. Disruption of adipocyte FAK in mice or in 3T3 L1 cells decreases adipocyte survival. Adipocyte-specific FAK knockout mice display impaired adipose tissue expansion and insulin resistance on prolonged metabolic stress from a high-fat diet or when crossed on an obese db/db or ob/ob genetic background. Treatment of these mice with a PPARγ agonist does not restore adiposity or improve insulin sensitivity. In contrast, inhibition of apoptosis, either genetically or pharmacologically, attenuates adipocyte death, restores normal adiposity and improves insulin sensitivity. Together, these results demonstrate that FAK is required for adipocyte survival and maintenance of insulin sensitivity, particularly in the context of adipose tissue expansion as a result of caloric excess.

Revascularización mediante aterectomía direccional en isquemia crítica de miembro inferior. Primera experiencia ecuatoriana / Revascularization through directional atherectomy in critical inferior limb ischemia: first Ecuadorian experience

Introducción: La aterectomía direccional es una técnica mínimamente invasiva que puede ser utilizada para evitar la amputación de los miembros inferiores en caso de isquemia crítica. Caso: Se presenta la primera experiencia en Ecuador, realizada en una paciente diabética e insuficiente renal. Este procedimiento logró repermeabilizar el segmento femoropoplíteo izquierdo con éxito. Discusión: La aterectomía direccional es una alternativa eficaz, mínimamente invasiva, para evitar la amputación de miembros inferiores en pacientes isquemia crítica y múltiples comorbilidades. Palabras clave: Aterectomía, isquemia, miembro inferior, pie diabético.

Introduction: Directional atherectomy is a minimally invasive technique that can be used to avoid lower limb amputation due to critical ischemia. Case report: This is the first experience in Ecuador, using this technique in a diabetic patient with chronic renal failure. The procedure achieved successful recanalization of the left femoropopliteal artery. Discusion: Directional atherectomy seems to be an alternative to avoid limb amputation in patients with critical ischemia and several comorbidities.

Comparing nucleic acid lateral flow and electrochemical genosensing for the simultaneous detection of foodborne pathogens.

Due to the increasing need of rapid tests for application in low resource settings, WHO summarized their ideal features under the acronym ASSURED (Affordable, Sensitive, Specific, User-friendly, Rapid and Robust, Equipment-free, Delivered to those who need it). In this work, two different platforms for the rapid and simultaneous testing of the foodborne pathogens E. coli O157:H7 and Salmonella enterica, in detail a nucleic acid lateral flow and an electrochemical magneto-genosensor are presented and compared in terms of their analytical performance. The DNA of the bacteria was amplified by polymerase chain reaction using a quadruple-tagging set of primers specific for E. coli eaeA (151bp) and Salmonella enterica yfiR (375bp) genes. During the amplification, the amplicons were labelled at the same time with biotin/digoxigenin or biotin/fluorescein tags, respectively. The nucleic acid lateral flow assay was based on the use of streptavidin gold nanoparticles for the labelling of the tagged amplicon from E. coli and Salmonella. The visual readout was achieved when the gold-modified amplicons were captured by the specific antibodies. The features of this approach are discussed and compared with an electrochemical magneto-genosensor. Although nucleic acid lateral flow showed higher limit of detection, this strategy was able to clearly distinguish positive and negative samples of both bacteria being considered as a rapid and promising detection tool for bacteria screening.

Eficacia de terapia fotodinámica como complemento de terapia convencional periodontal versus terapia convencional en el tratamiento de pacientes adultos con periodontitis crónica: una revisión sistemática con metaanálisis / Effectiveness photodynamic therapy complementing conventional periodontal therapy versus conventional therapy in the treatment of adult patients with chronic periodontitis: a systematic review with meta-analyses

Se evaluó la eficacia de la terapia fotodinámica como complemento de terapia periodontal convencional comparándola con la terapia convencional en el tratamiento de sacos periodontales en pacientes adultos con periodontitis crónica basándose en ensayos clínicos comprendidos entre los años 2010 y 2015, para determinar si su uso otorga mejores resultados para el tratamiento de esta enfermedad. Se seleccionaron ensayos clínicos aleatorios prospectivos, aleatorizados o no aleatorizados, controlados y no controlados que permitieron la comparación entre el tratamiento convencional y la terapia fotodinámica, con un grupo en el cual se utilizó sólo la terapia convencional. Los datos de los ensayos clínicos fueron ingresados al software Review Manager®. Se realizaron tres metaanálisis para las variables: Nivel de inserción clínica (NIC) y profundidad de sondaje (PS), el test de I2 fue utilizado para medir la heterogeneidad del estudio y posteriormente un análisis de sensibilidad para determinar los estudios heterogéneos. Se pudieron analizar 7 estudios, con un total de 186 pacientes, quienes fueron controlados 3 meses post tratamiento. Se utilizó la diferencia de medias, un intervalo de confianza de 95 % para medir el NIC y PS. A los 3 meses, no se encontró diferencias significativas en NIC (p= 0,93) y PS (p= 0,71). Conclusión: La terapia fotodinámica en complementación a la terapia convencional no otorga mejor resultado clínico ni estadístico comparado con la terapia convencional al evaluar el nivel de inserción clínica. Al evaluar la profundidad de sondaje es recomendable la utilización de terapia convencional sola.

The efficacy of photodynamic therapy as an adjunct to conventional periodontal therapy evaluated by comparing with conventional therapy alone in the treatment of periodontal pockets in adult patients with chronic periodontitis based on clinical trials between 2010 and 2015, to determine if its use can provide better results for treating this disease. Prospective randomized clinical trials and randomized clinical trials or non-randomized, controlled and uncontrolled that allowed comparison between a group which was applied to conventional therapy and photodynamic therapy, a similar group was selected to which you He applied only conventional therapy. Data from clinical trials entered into Review Manager®. Three meta-analyzes for the variables analyzed were performed: Level clinical attachment (NIC) and probing depth (PS), the test of I2 was used to measure the heterogeneity of the study and then a sensitivity analysis to determine which studies awardedheterogeneity. As results, seven studies analyzed in 186 patients who underwent treatment at least controlled within 3 months post treatment. The mean difference was used, a confidence interval of 95 % to measure the NIC and PS. At 3 months, no significant differences in NIC (p= 0.93) and PS (p= 0.71). In conclusion, the photodynamic therapy complementary to conventional therapy does not provide better clinical or statistical results compared with conventional therapy to evaluate the clinical attachment level. In assessing probing depth, is advisable to use conventional therapy alone.

Reduced Frequencies and Activation of Regulatory T Cells After the Treatment of HIV-1-Infected Individuals with the CCR5 Antagonist Maraviroc Are Associated with a Reduction in Viral Loads Rather Than a Direct Effect of the Drug on Regulatory T Cells.

Regulatory T cells (Tregs) play an important role in the pathogenesis of HIV-1 infection and they frequently express the chemokine receptor CCR5. We therefore investigated whether antiretroviral treatment with the CCR5 antagonist Maraviroc affected Tregs in chronically HIV-1-infected individuals. HIV-1-infected patients with high viral loads had elevated frequencies of activated Tregs in the peripheral blood compared with healthy controls. In patients successfully treated with antiretroviral drugs (undetectable viral loads), the frequency and the activation status of Tregs were comparable with healthy controls without any specific effect related to the treatment with Maraviroc. These results indicate that the control of viral replication in general rather than a direct binding of Maraviroc to CCR5-positive Tregs influences Treg responses in successfully treated chronically HIV-1-infected individuals.

JAK2 promotes brown adipose tissue function and is required for diet- and cold-induced thermogenesis in mice.

AIMS/HYPOTHESIS: Non-shivering thermogenesis in adipose tissue can be activated by excessive energy intake or following cold exposure. The molecular mechanisms regulating this activation have not been fully elucidated. The Janus kinase (JAK) - signal transducer and activator of transcription (STAT) pathway mediates the signal transduction of numerous hormones and growth factors that regulate adipose tissue development and function, and may play a role in adaptive thermogenesis. METHODS: We analysed mRNA and protein levels of uncoupling protein 1 (UCP1) and JAK2 in different adipose depots in response to metabolic and thermal stress. The in vivo role of JAK2 in adaptive thermogenesis was examined using mice with adipocyte-specific Jak2 deficiency (A-Jak2 KO). RESULTS: We show in murine brown adipose tissue (BAT) that JAK2 is upregulated together with UCP1 in response to high-fat diet (HFD) feeding and cold exposure. In contrast to white adipose tissue, where JAK2 was dispensable for UCP1 induction, we identified an essential role for BAT JAK2 in diet- and cold-induced thermogenesis via mediating the thermogenic response to ß-adrenergic stimulation. Accordingly, A-Jak2 KO mice were unable to upregulate BAT UCP1 following a HFD or after cold exposure. Therefore, A-Jak2 KO mice were cold intolerant and susceptible to HFD-induced obesity and diabetes. CONCLUSIONS/INTERPRETATION: Taken together, our results suggest that JAK2 plays a critical role in BAT function and adaptive thermogenesis. Targeting the JAK-STAT pathway may be a novel therapeutic approach for the treatment of obesity and related metabolic disorders.