Direct characterization of circulating DNA in blood plasma using μLAS technology

Circulating cell-free DNA (cfDNA) is a powerful cancer biomarker for establishing targeted therapies or monitoring patients' treatment. However, current cfDNA characterization is severely limited by its low concentration, requiring the extensive use of amplification techniques. Here we report that the μLAS technology allows us to quantitatively characterize the size distribution of purified cfDNA in a few minutes, even when its concentration is as low as 1 pg/μL. Moreover, we show that DNA profiles can be directly measured in blood plasma with a minimal conditioning process to speed up considerably speed up the cfDNA analytical chain

HIV-1 suppression and durable control by combining single broadly neutralizing antibodies and antiretroviral drugs in humanized mice

Effective control of HIV-1 infection in humans is achieved using combinations of antiretroviral therapy (ART) drugs. In humanized mice (hu-mice), control of viremia can be achieved using either ART or by immunotherapy using combinations of broadly neutralizing antibodies (bNAbs). Here we show that treatment of HIV-1–infected hu-mice with a combination of three highly potent bNAbs not only resulted in complete viremic control but also led to a reduction in cell-associated HIV-1 DNA. Moreover, lowering the initial viral load by coadministration of ART and immunotherapy enabled prolonged viremic control by a single bNAb after ART was withdrawn. Similarly, a single injection of adeno-associated virus directing expression of one bNAb produced durable viremic control after ART was terminated. We conclude that immunotherapy reduces plasma viral load and cell-associated HIV-1 DNA and that decreasing the initial viral load enables single bNAbs to control viremia in hu-mice

Inflammatory Cytokine Expression Is Associated with Chikungunya Virus Resolution and Symptom Severity

The Chikungunya virus infection zones have now quickly spread from Africa to parts of Asia, North America and Europe. Originally thought to trigger a disease of only mild symptoms, recently Chikungunya virus caused large-scale fatalities and widespread economic loss that was linked to recent virus genetic mutation and evolution. Due to the paucity of information on Chikungunya immunological progression, we investigated the serum levels of 13 cytokines/chemokines during the acute phase of Chikungunya disease and 6- and 12-month post-infection follow-up from patients of the Italian outbreak. We found that CXCL9/MIG, CCL2/MCP-1, IL-6 and CXCL10/IP-10 were significantly raised in the acute phase compared to follow-up samples. Furthermore, IL-1β, TNF-α, Il-12, IL-10, IFN-γ and IL-5 had low initial acute phase levels that significantly increased at later time points. Analysis of symptom severity showed association with CXCL9/MIG, CXCL10/IP-10 and IgG levels. These data give insight into Chikungunya disease establishment and subsequent convalescence, which is imperative to the treatment and containment of this quickly evolving and frequently re-emerging disease

Ertüchtigung der Flachschleifmaschine ELB SF 30

Aufbauend auf dem Praktikumsbericht wird in der vorliegenden Bachelorarbeit, das Thema „Ertüchtigung der Flachschleifmaschine ELB“ weiter behandelt. Es wird dabei speziell die Einführung eines neuen Bedienpanels an der Maschine konkretisiert. Zu Beginn erfolgt die Auswahl eines geeigneten Panels, welches zu der vorhandenen Steuerung kompatibel ist. Daraufhin folgt die Vorstellung der Visualisierungssoftware, sowie der Software zum Testen der projektierten Oberfläche des Bediengerätes. Im weiteren Verlauf finden sich Erläuterungen zur Programmierung des Panels, bezogen auf das ELB-Projekt wieder. Das letzte Kapitel bezieht sich auf die konkrete SPS-Lösung zwei ausgewählter Steuerungsfunktionen, zum einen die Magnetsteuerung und zum anderen die Querantriebsteuerung

The effectiveness of different rat IgG subclasses as IgE-blocking antibodies in the Rat Basophil Leukemia cell model

The degranulation of mast cells in an allergic response is initiated by the aggregation of high-affinity IgE receptors (Fc epsilon RI) by IgE and antigen. Recently it has been shown that such degranulation can be inhibited by cross-linking FceRI and low-affinity IgG receptors (Fc gamma RII) which are also expressed by mast cells. The ability of various monoclonal antibodies to block the degranulation of rat basophil leukaemia (RBL) cells sensitized with IgE antidinitrophenyl (DNP) antibodies has been investigated. Sensitized cells were challenged with immune complexes formed using varying concentrations of antigen, and of both high- and low-valency antigen. It is reported here that rat IgG1 antibodies, which are associated in the rat with a Th1-type response, act as highly effective blocking antibodies over a wide concentration range. Rat IgG2a antibodies, which are associated with a Th2-type response, were able only to inhibit degranulation when immune complexes were formed with very low concentrations of high-valency antigen (DNP32-HSA). Under these conditions, some inhibitory activity was seen with high-affinity murine IgA anti-DNP but not with low-affinity rat IgG2b anti-DNP antibody-containing immune complexes. In addition to this inhibitory activity, IgG2a antibodies were shown to be capable of inducing degranulation of cells via unoccupied FceRI. These results demonstrate that blocking activity may arise via both inhibitory receptors and by masking of antigen

Engine combustion network (Ecn) : characterization and comparison of boundary conditions for different combustion vessels

The Engine Combustion Network (ECN) is a worldwide group of institutions using combustion vessels and/or performing computational fluid dynamics (CFD) simulation, whose aim is to advance the state of spray and combustion knowledge at engine-relevant conditions. A key activity is the use of spray chamber facilities that operate at high-temperature, high-pressure conditions typical of diesel combustion, which are operated at specific target conditions in order to leverage research capabilities and advanced diagnostics of all ECN participants. The first target condition, called "Spray A," has been defined with detailed ambient (900 K, 60 bar, 22.8 kg/m(3), 15% oxygen) and injector (common rail, 1500 bar, KS1.5/86 nozzle, 0.090-mm orifice diameter, n-dodecane, 363 K) conditions. Establishing and improving these experimental boundary conditions in unique facilities throughout the world represents a major step forward in the establishment of high-quality, quantitative data sets for engine spray combustion. This paper is a review of the methodology to characterize and control the ambient and fuel-injector boundary conditions (e.g., temperature, pressure, composition) as offered by six different participating institutions of the ECN, each targeting the Spray A conditions and quantifying experimental uncertainty. Constant-pressure flow (CPF) and constant-volume preburn (CVP) chambers with various ambient gas composition are compared for the first time. Experimental diagnostics include the use of fast-response, radiation-corrected thermocouples for spatially resolved gas and fuel-injector temperature, laser-induced phosphorescence for surface temperature, and high-speed transducers for pressure. With guidance about the uncertainty and variation that exists between facilities, simplified models are then employed to understand how these boundary condition variations may affect aspects of spray combustion. Ambient gas and fuel temperature effects on liquid- and vapor-phase penetration are examined with established one-dimensional models. Chemical kinetics modeling in single- or multi-zone reactors is used to predict the influence of different preburn environments on the major and minor species present in the ambient gas at the start of injection, and their subsequent effect on spray ignition. This review article provides recognition of the challenge in creating well-controlled high-temperature, high-pressure environments, and identifies which boundary condition variations are expected to have the highest impact on spray combustion

The Src family kinase, Lyn, suppresses osteoclastogenesis in vitro and in vivo

c-Src kinase is a rate-limiting activator of osteoclast (OC) function and Src inhibitors are therefore candidate antiosteoporosis drugs. By affecting αvβ3 and macrophage-colony stimulating factor (M-CSF)-induced signaling, c-Src is central to osteoclast activity, but not differentiation. We find Lyn, another member of Src family kinases (SFK) is, in contrast, a negative regulator of osteoclastic bone resorption. The absence of Lyn enhances receptor activator of NF-κB ligand (RANKL)-mediated differentiation of osteoclast precursors without affecting proliferation and survival, while its overexpression decreases osteoclast formation. In further contrast to c-Src, Lyn deficiency does not impact the activity of the mature cell. Reflecting increased osteoclast development in vitro, Lyn−/− mice undergo accelerated osteoclastogenesis and bone loss, in vivo, in response to RANKL. Mechanistically, Lyn forms a complex with receptor activator of NF-κB (RANK), the tyrosine phosphatase, SHP-1, and the adapter protein, Grb2-associated binder 2 (Gab2). Upon RANKL exposure, Gab2 phosphorylation, JNK, and NF-κB activation are enhanced in Lyn−/− osteoclasts, all critical events in osteoclast development. We therefore establish that Lyn regulates osteoclast formation and does it in a manner antithetical to that of c-Src. The most pragmatic aspect of our findings is that successful therapeutic inhibition of c-Src, in the context of the osteoclast, will require its stringent targeting

Tratamento cirúrgico da ginecomastia com pedículos lateral e medial

Ginecomastia é o aumento da mama masculina que pode acometer até 65% dos indivíduos deste sexo na fase infanto-puberal, compreendida entre 13 e 16 anos. Tem como principais causas hepatite ou cirrose hepática, carcinoma ou doenças inflamatórias pulmonares crônicas, carcinomas ou disfunções testiculares, tumores glandulares (pituitária, supra-renal), alterações dos níveis séricos de testosterona, síndromes genéticas (síndrome de Klinefelter, p.ex.), uso de drogas como heroína, maconha ou anabolizantes e hanseníase. Podemos classificar a ginecomastia quanto ao volume, quanto aos tecidos que a compõem (gordurosa ou pseudoginecomastia, glandular e mista), ou quanto ao tratamento necessário para sua correção cirúrgica (pequena, moderada e grave). O tratamento das formas mais graves de ginecomastia é muito diferente daquele aplicado às formas mais suaves, pois nas formas graves, além da ressecção dos tecidos gorduroso e glandular, existe a necessidade de ressecção da pele em excesso e o reposicionamento do complexo aréolo-mamilar. O objetivo deste trabalho é descrever uma técnica cirúrgica específica para estes pacientes portadores de formas graves de ginecomastia, através de dois pedículos dermogordurosos, um lateral e um medial, com aproximadamente 2cm de espessura, mantendo assim a nutrição do complexo aréolo-mamilar. Esses pedículos são delimitados entre as bissetrizes dos quadrantes súpero-lateral e ínfero-lateral, e súpero-medial e ínfero-medial, tendo o mamilo como vértice. Na área de pele excessiva periareolar, obtida através do pinçamento interdigital, é realizada a desepidermização dos pedículos lateral e medial e ressecção de toda pele e tecido celular subcutâneo até a fáscia peitoral nas regiões superior e inferior aos pedículos; a síntese é realizada em dois planos, sendo periareolar a cicatriz resultante. Foram operados com esta técnica vinte pacientes com forma grave de ginecomastia, com média etária de 23,3 anos; sendo seis pacientes da raça negra. O bom posicionamento do complexo aréolo-mamilar e uma cicatriz periareolar resultante, bem como a retirada de conteúdo suficiente, foram as principais vantagens observadas. Como complicações, tivemos assimetria das placas aréolo-mamilares em dois casos, nos quais havia acentuada diferença entre os dois lados na avaliação pré-operatória; cicatrização hipertrófica em um paciente da raça negra, cuja cicatriz foi atenuada com injeções intracicatriciais de triancinolona; necrose parcial de aréola em um caso, cuja ferida cicatrizou por segunda intenção, dispensando qualquer tratamento local posterior; deiscência de sutura periareolar em um caso, no qual foi feita a ressutura, com bom resultado, e quatro pacientes apresentaram coleção sero-hemática subcutânea, que foram drenadas e não apresentaram recidiva

Tregs and allergic disease

Allergic diseases such as asthma, rhinitis, and eczema are increasing in prevalence and affect up to 15% of populations in Westernized countries. The description of Tregs as T cells that prevent development of autoimmune disease led to considerable interest in whether these Tregs were also normally involved in prevention of sensitization to allergens and whether it might be possible to manipulate Tregs for the therapy of allergic disease. Current data suggest that Th2 responses to allergens are normally suppressed by both CD4(+)CD25(+) Tregs and IL-10 Tregs. Furthermore, suppression by these subsets is decreased in allergic individuals. In animal models, Tregs could be induced by high- or low-dose inhaled antigen, and prior induction of such Tregs prevented subsequent development of allergen sensitization and airway inflammation in inhaled challenge models. For many years, allergen-injection immunotherapy has been used for the therapy of allergic disease, and this treatment may induce IL-10 Tregs, leading to both suppression of Th2 responses and a switch from IgE to IgG4 antibody production. Improvements in allergen immunotherapy, such as peptide therapy, and greater understanding of the biology of Tregs hold great promise for the treatment and prevention of allergic disease