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The microwave temperature profiler (MTP), an airborne passive microwave radiometer, measures radiances, recorded as counts and calibrated to brightness temperatures, in order to estimate temperature profiles around flight altitude. From these data, quantities such as potential temperature gradients and static stability, indicating the state of the atmosphere, can be derived and used to assess important dynamical processes (e.g., gravity waves or stability assessments). DLR has acquired a copy of the MTP from NASA–JPL, which was designed as a wing-canister instrument and is deployed on the German High Altitude LOng range research aircraft (HALO). For this instrument a thorough analysis of instrument characteristics has been made in order to correctly determine the accuracy and precision of MTP measurements. Using a laboratory setup, the frequency response function and antenna diagram of the instrument were carefully characterized. A cold chamber was used to simulate the changing in-flight conditions and to derive noise characteristics as well as reliable calibration parameters for brightness temperature calculations, which are compared to those calculated from campaign data. The MTP shows quite large changes in the instrument state, imposing considerable changes in calibration parameters over the course of a single measurement flight; using a built-in heated target for calibration may yield large errors in brightness temperatures due to a misinterpretation of the measured absolute temperature. Applying the corrections presented herein to the calibration parameter calculations, the measurement noise becomes the dominant source of uncertainty and it is possible to measure the brightness temperatures around flight level (closely related to the absolute temperature close to the instrument) with a precision of 0.38 K. Furthermore, radiative transfer simulations, using the Py4CAtS package in a pencil-beam approach, indicate that the altitude range of the sensitivity of the MTP instrument can be increased by applying a modified measurement strategy. This is the first time such an extensive characterization of an MTP instrument, including a thorough calibration strategy assessment, has been published. The presented results, relevant for the wing-canister design of the MTP instrument, are important when processing MTP data: knowledge of the relevant uncertainties and instrument characteristics is essential for retrieval setup and is mandatory to correctly identify and interpret significant atmospheric temperature fluctuations.

Tuberculosis remains a widespread and deadly infectious disease, especially in the developing world. Despite the availability of effective antibiotics, treatment of the infection is challenging due to the extreme persistence of the causative pathogen Mycobacterium tuberculosis. Recent studies have shown that M. tuberculosis can form multicellular colonies, referred to as biofilms. A biofilm-like lifestyle provides bacteria with increased tolerance to external stress factors, such as physical forces and chemicals, and creates persistence-favoring environments. One of the hallmark features of biofilms is reduced susceptibility to antibiotics. M. tuberculosis is primarily an intracellular pathogen, habituating maturation-stalled phagosomes of macrophages. Biofilm-like communities of mycobacteria are suggested to reside in the extracellular environment of the host, such as surfaces of alveolar cavities and necrotic granulomas. One of the key components of bacterial communities is the extracellular matrix. Through the lessons learned from other biofilm-forming species, it is known that the matrix generally consists of polysaccharides, proteins, extracellular DNA and lipids. Components of the mycobacterial biofilm matrix have not been explicitly studied, although several studies have indicated the presence of mycolic acids and cellulose. This thesis investigates the role of cellulase and mycobacterial pilin in biofilm formation by over-expressing celA1 (MMAR_0107) and mtp (MMAR_3485) genes in Mycobacterium marinum. Both over-expression strains retained capability for planktonic growth, but their biofilm-like growth was disrupted as observed by confocal laser scanning microscopy (CLSM). Formation of rope-like ‘cording’ structures was disrupted in mtp over-expressing strain, and increased aggregation of bacteria was observed. Biofilm formation of celA1 over-expression strain was severely disrupted, implying that a substrate of CelA1-cellulase is an important structural component of the biofilm of M. marinum. Amyloids are being increasingly discovered from biofilms of various species. Being highly resistant proteinaceous fibers, amyloids exhibit cytotoxicity in multiple human diseases. In contrast to pathological amyloids, functional amyloids, known for both mammals and bacteria, are utilized for cellular functions. Due to the analogy of M. tuberculosis pili (MTP) with known functional amyloids curli pili of E. coli, MTP were considered as candidates for biofilm-constituting amyloid in mycobacteria. Our experiments and recent investigation by other researchers could not confirm the amyloidogenicity of MTP and therefore we applied ab initio amyloid identification approach We identified a novel mycobacterial amyloid candidate as a putative protein with a SIR2-like domain (MMRN_51880). Based on in silico analysis, the protein had aggregative sequences. The protein was produced in both biofilm and planktonic culture. The function of the protein has not been annotated yet, but the presence of SIR2-domain suggests that the protein belongs to protein deacetylases. Further studies are required for the verification of a new candidate as a novel mycobacterial functional amyloid. Overall, this study contributes to our scarce understanding of mycobacterial biofilm matrix components which could be further investigated for antibiofilm drug development purposes. The antibiofilm therapy could allow us to enhance the effect of existing antibiotics and drastically shorten the duration of antimycobacterial treatment, contributing to the eradication of the tuberculosis pandemic. Tuberkuloosi on yhä laajasti esiintyvä ja tappava infektiotauti eteenkin kehittyvissä maissa. Vaikka toimiva hoito on saatavilla, taudin parantaminen on edelleen haastavaa. Aiheuttajapatogeeni Mycobacterium tuberculosis on kehittynyt äärimäisen vastustuskykyiseksi ihmisen immuunisuojaa ja antibiootteja vastaan. Hiljattain M. tuberculosis -bakteerin on osoitettu muodostavan monisoluisia yhteisöjä eli biofilmejä. Biofilminmuodostus on tunnettu ilmiö bakteereilla, ja sen tiedetään edesauttavan bakteerien selviytymistä vaikeissa ympäristöolosuhteissa. Biofilminkaltainen elintapa on tunnettu eteenkin korkeasta antibiootinsietokyvystään. M. tuberculosis -bakteeri on ensisijaisesti solunsisäinen patogeeni, joka on erikoistunut selviytymään makrofageissa. Biofilminkaltaista elämänmuotoa ajatellaan puolestaan esiintyvän isäntäsolujen ulkopuolella. Biofilmien keskeinen tekijä on soluväliaine. Bakteerien biofilmin soluväliaine koostuu yleisimmin polysakkarideista, solunulkopuolisesta DNA:sta, proteiineista ja lipideistä. Mykobakteerien biofilmien soluväliaineen komponentteja ei tunneta täysin, mutta muutamat aiheesta tehdyt tutkimukset ovat osoittaneet mm. mykolihappojen ja selluloosan osallisuuden. Tässä tutkielmassa olen tarkastellut sellulaasin ja mykobakteerien piliinin vaikutusta biofilminmuodostukseen. Tähän tarkoitukseen valmistin celA1 (MMAR_0107) ja mtp (MMAR_3485) geenejä ilmentäviä kantoja käyttäen Mycobacterium marinum -lajia. Molemmat yliekspressiokannat säilyttivät kyvyn planktoniseen kasvuun. Yliekspressiokantojen biofilminkaltainen kasvu oli kuitenkin huomattavasti villityypistä poikkeavaa konfokaalimikroskopialla tarkasteltuna. Mykobakteereille tyypilliset nauhamaiset rakenteet ilmentyivät mtp-yliekspressiokannassa aggregoituneena solumassana. Biofilmimuodostus celA1-yliekspressiokannassa oli huomattavasti heikentynyt viittaen sellulaasin substraatin olevan tärkeä rakenteellinen osa M. marinum:n -biofilmiä. Amyloideja tunnistetaan erilaisten biofilmien soluväliaineesta enenevässä määrin. Amyloidit ovat erittäin kestäviä proteiinikuituja, joiden sytotoksisia ominaisuuksia on tunnistettu ihmisen tautien yhteydessä. Funktionaaliset amyloidit, toisin kuin tauteja aiheuttavat, ovat tarpeellisia solujen toiminnoissa. M. tuberculosis -pilukset (MTP) ovat osoittautuneet aiemmissa tutkimuksissa samankaltaisiksi kuin tunnetut funktionaaliset amyloidit – E. coli:n curli pilukset. Tämän vuoksi ajatellaan, että MTP saattaa muodostaa amyloideja mykobakteereissa, mutta ilmiötä ei ole vielä osoitettu. Tämän vuoksi sovelsimme suoraa amyloidien tunnistusmenetelmää. Tunnistettu monomeeri osoittautui ennustetuksi proteiiniksi (MMRN_51880), jossa on SIR2-kaltainen domeeni. Sekvenssianalyysin perusteella tunnistetulla proteiinilla on amyloidin muodostusta mahdollistavia sekvenssejä. Proteiinia tuotetaan todennäköisesti sekä biofilmisen että planktonisen kasvun aikana. Proteiinin merkitystä ei tunneta, mutta sen sisältämä SIR2-kaltainen domeeni viittaa proteiinien deasetylaatioon. Jatkotutkimuksien avulla olisi mahdollista selvittää, muodostaako tunnistettu proteiini mykobakteerien amyloideja. Yleisesti ottaen uusi tieto mykobakteerien biofilmeistä on tärkeää, koska se mahdollistaa biofilminvastaisten lääkkeiden kehityksen. Yhdistettynä olemassa oleviin antibiootteihin hoitostrategia voisi tehostaa merkittävästi käytössä olevien antibioottien tehoa ja lyhentää antibioottikuurien kestoa helpottaen näin tuberkuloosiepidemian hävittämistä.

Yen-Michael S Hsu,1 Thorsten Haas,2 Melissa M Cushing1 1Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY, USA; 2Department of Anesthesia, University Children's Hospital Zurich, Zurich, Switzerland Abstract: Massive transfusion protocols (MTPs) are established to provide rapid blood replacement in a setting of severe hemorrhage. Early optimal blood transfusion is essential to sustain organ perfusion and oxygenation. There are many variables to consider when establishing an MTP, and studies have prospectively evaluated different scenarios and patient populations to establish the best practices to attain improved patient outcomes. The establishment and utilization of an optimal MTP is challenging given the ever-changing patient status during resuscitation efforts. Much of the MTP literature comes from the trauma population, due to the fact that massive hemorrhage is the leading cause of preventable trauma-related death. As we come to further understand the positive and negative clinical impacts of transfusion-related factors, massive transfusion practice can be further refined. This article will first discuss specific MTPs targeting different patient populations and current relevant international guidelines. Then, we will examine a wide selection of therapeutic products to support MTPs, including newly available products and the most suitable of the traditional products. Lastly, we will discuss the best design for an MTP, including ratio-based MTPs and MTPs based on the use of point-of-care coagulation diagnostic tools. Keywords: hemorrhage, MTP, antifibrinolytics, coagulopathy, trauma, ratio, logistics, guidelines, hemostatic

Background:Arthrodesis of the ipsilateral hallux metatarsophalangeal (MTP) and interphalangeal (IP) joints may be required for severe arthritis or deformity at both joints. The purpose of this study was to review outcomes of ipsilateral first MTP and IP joint arthrodesis.Methods:Twenty feet were identified, for which the diagnosis was rheumatoid arthritis in 14, failed hallux valgus surgery in 5, and hallux rigidus in 1. The IP arthrodesis was performed first in 6 feet; MTP first in 8 feet; and both joints simultaneously in 6 feet. Median follow-up was 28 months (range 12-94). Medical records and radiographs were reviewed. American Orthopaedic Foot & Ankle Society (AOFAS) score and patient satisfaction were determined.Results:Although all of the MTP arthrodeses healed, 8 of 20 feet (40%) failed to heal at the IP arthrodesis. The rate of IP nonunion was 17% (1/6) with IP arthrodesis first, 50% (4/8) with MTP arthrodesis first, and 50% (3/6) with simultaneous arthrodesis. Four of 8 IP nonunions were symptomatic. Subsequent surgery was required in 11 feet (55%), including repair of IP nonunion in 3 feet, hardware removal in 4, revision MTP malunion in 2, wound debridement in 1, and soft tissue reconstruction in 1. Median hallux AOFAS score for the cohort increased from 25 to 68. Eighteen feet resulted in patients who were very satisfied or satisfied with minor reservations. Neither AOFAS score nor satisfaction trended toward association with IP union.Conclusion:Ipsilateral arthrodesis of the hallux MTP and IP joints was challenging because of high rates of reoperation and IP nonunion, the latter of which was likely related to increased mechanical stress on the IP joint with immobilization of the MTP joint. Despite the high IP nonunion rate, IP nonunion did not predict patient-reported outcome. Fibrous ankylosis was an acceptable clinical outcome in many cases.Level of Evidence:Level IV, case series.

The support of 5G verticals service requires todesign an efficient Mobile Transport and Computing Platformwhere transport, mobile and MEC must interact effectively. Inthis paper, a novel architecture is proposed providing itsmapping on ETSI NFV. Two relevant use cases, such asautomotive and cloud robotics are presented to assess the novelarchitecture. This work has been partially funded by the EU H2020 5G-Transformer Project (grant no. 761536).

B cells (BC) play a critical role in systemic lupus erythematosus (SLE). BC depletion therapy still remains an attractive option, despite the disappointing results of randomized controlled trials (RTCs).Twelve patients with SLE [3 males, mean age 43.8 yrs (25-55)] with severe multiorgan involvement all including kidney (3 patients with Class IV, 4 with Class III/V and 5 with Class V, according to the International Society of Nephrology/Renal Pathology Society glomerulonephritis classification), skin lesions [10], severe polyarthralgias with arthritis [10], polyserositis [2], and lymphadenopathy [5] have been prospectively treated with an intensified B cell depletion therapy (IBCDT) protocol due to their resistance or intolerance to previous therapy (six cases) or as a front line immunosuppressive treatment in 6 women with unsatisfactory therapeutic compliance or as a specific request of a short-time immunosuppression for gestational perspectives.Rituximab (RTX) 375 mg/sm on days 1, 8, 15, 22, and 2 more doses after 1 and 2 months, associated with 2 IV administrations of 10mg/kg of cyclophosphamide and 3 methylprednisolone pulses (15mg/kg) followed by oral prednisone (0.8 mg/kg/day, rapidly tapered to 5mg/day by the end of the 3rd month after RTX). No further immunosuppressive maintenance therapy has been given.Patients had been followed-up for a mean of 44.5 (24-93)months. Significant decreases (p0.05) were found in the levels of ESR (baseline mean value: 55.0mm; 3 months: 36; end of follow-up: 13), anti-dsDNA antibodies (baseline: 185 U; 3 months: 107; end of follow-up: 15), and proteinuria (baseline: 4.9 g/24h; 3 months: 0.97; end of follow-up: 0.22). C4 values (baseline 11 mg/dl) significantly increased (p0.05) after 3 months (22 mg/dl) and at the end of the follow-up (20mg/dl). Of the 12 patients, 9 (75%) have remained well after one cycle of IBCDT, with no flare (mean 51.6 months [25-93]). Three patients relapsed after 36, 41, and 72 months, respectively. Following re-treatment, they again showed complete remission over 18-48 months of observation.A promising role of RTX in an intensified protocol of induction therapy can be envisaged in patients for whom avoiding immunosuppressive maintenance therapy and sparing steroids are particularly appealing. Moreover, our data confirm in one of the longest follow-up available, the opportunity to reconsider the regimens of BL depletion in the treatment of the most severe or refractory forms of SLE despite the disappointing results of RCTs.

Renal failure in obstetrics is rare but important complication, associated with significant mortality and long term morbidity.1,2 It includes acute renal failure due to obstetrical complications or due to deterioration of existing renal disease. AIMS AND OBJECTIVES: To evaluate the etiology and outcome of renal failure in obstetric patients. METHODS: We prospectively analyzed 30 pregnant and puerperal women with acute renal failure or pre-existing renal disease developing renal failure during pregnancy between November 2007 to sep-2009. Patients who presented/developed ARF during the hospital stay were included in this study. RESULTS: Among 30 patients, mean age was 23 years and 33 years age group. 12 cases (40%) patients were primigravidae and 9(30%) patients were multigravidae and 9 cases (30%) presented in post-partum period. Eighteen cases (60%) with ARF were seen in third trimester, followed by in postpartum period 9 cases (30%). Most common contributing factors to ARF were Pre-eclampsia, eclampsia and HELLP syndrome 60%, sepsis 56.6%, post abortal ARF 10%. DIC 40%. Haemorrhage as the aetiology for ARF was present 46%, APH in 20% and PPH in 26.6%. The type of ARF was renal in (63%) and prerenal (36%); Oliguric seen in 10 patients (33%) and high mortality (30%). Among the 20 pregnant patients with ARF, The average period of gestation was 33±2 weeks (30 -36 weeks), 5 cases (25%) presented with intrauterine fetal demise and 18 cases (66%) had preterm vaginal delivery and 2 cases (10%) had induced abortion. And the average birth weight was 2±0.5 kg (1.5 kg). Eight cases (26%) required dialysis. 80% of patients recovered completely of renal functions. 63% patients recovered without renal replacement therapy whereas 17% required dialysis. the maternal mortality was 20%, the main reason for mortality was septic shock and multi organ dysfunction (66%). CONCLUSION: ARF related pregnancy was seen commonly in the primigravidae and in the third trimester, the most common reasons were pregnancy induced hypertension, HELLP syndrome and obstetric haemorrhage and resulted in high risk condition for fetal and maternal mortality. The most effective measures still remain the careful prevention and the aggressive management of the obstetric complications. Ideal care for women with acute renal failure in pregnancy or post-partum requires a multidisciplinary approach that includes maternal-fetal medicine, critical care medicine, nephrology and neonatology specialties.

Among all the metabolites present in the plasma, lipids, mainly triacylglycerol and diacylglycerol, show extensive circadian rhythms. These lipids are transported in the plasma as part of lipoproteins. Lipoproteins are synthesized primarily in the liver and intestine and their production exhibits circadian rhythmicity. Studies have shown that various proteins involved in lipid absorption and lipoprotein biosynthesis show circadian expression. Further, intestinal epithelial cells express circadian clock genes and these genes might control circadian expression of different proteins involved in intestinal lipid absorption. Intestinal circadian clock genes are synchronized by signals emanating from the suprachiasmatic nuclei that constitute a master clock and from signals coming from other environmental factors, such as food availability. Disruptions in central clock, as happens due to disruptions in the sleep/wake cycle, affect intestinal function. Similarly, irregularities in temporal food intake affect intestinal function. These changes predispose individuals to various metabolic disorders, such as metabolic syndrome, obesity, diabetes, and atherosclerosis. Here, we summarize how circadian rhythms regulate microsomal triglyceride transfer protein, apoAIV, and nocturnin to affect diurnal regulation of lipid absorption.

Dissertation, Rheinisch-Westfälische Technische Hochschule Aachen, 2018; Aachen 1 Online-Ressource (XVIII, 118 Seiten) : Illustrationen, Diagramme (2018). = Dissertation, Rheinisch-Westfälische Technische Hochschule Aachen, 2018, Industrial large-scale bioprocesses are often run in fed-batch operation mode. It provides several advantages compared to the conventional batch mode such as optimal cultivation conditions as well as enhanced product yields. In contrast, small-scale screening is often run in batch mode. However, the physiological conditions between both operation modes strongly vary and suboptimal strains might be selected from the screening process. In the past years, different small-scale fed-batch bioreactors have been developed. A novel screening tool represents the fed-batch microtiter plate, presented in this work. The system enables diffusive substrate release from a hydrogel into a culture well. Within this work, the microtiter plates and substrate release are characterized in detail. In a first step, the diffusion properties of a polyvinyl alcohol (PVA) and polyallylamine (PAAm) hydrogels were analyzed for feeding concentrations up to 700 g/L. For the PVA-glycerol and PAAm-glucose systems, maximum diffusion rates of 0.86 mg/h and 0.58 mg/h were determined, respectively. Moreover, the apparent lag-phase tlag was identified as key parameter regarding the hydrogel stability. In a second step, substrate diffusion through the hydrogel was calculated based on a hydrodynamic diffusion model. Moreover, a simulation model based on Fick’s diffusion laws was developed. Predictions about the diffusive mass flows were made and the local volume change of the hydrogel was calculated. As a result, substrate diffusion could be predicted up to +/- 20 % by means of the simulation model. In addition, water counter diffusion was identified as having considerable impact onto the hydrogel stability. For the PAAm hydrogel, a relative volume loss of 7.5 - 12.5 % per section was calculated as critical volume loss at feeding concentrations above 300 g/L. In addition, a maximum volume loss of 19 % after 0.8 h was observed for a feeding concentration of 700 g/L. To demonstrate the biological application of the novel fed-batch microtiter plate, cultivations experiments with E. coli BL21 (DE3) pRhotHi-2-Ec FbFP were performed. Here, carbon limited fed-batch conditions could be observed. Finally, experiments regarding the temperature distribution across conventional microtiter plates have been performed. A relative deviation of the temperature of 1.5 – 2.3 °C across the microtiter plate was determined., Published by Aachen

The plasma level of apolipoprotein B (apoB) is among the strongest risk factors for coronary artery disease. Microsomal triglyceride transfer protein (MTP) plays a key role in the lipidation of nascent apoB and the secretion of apoB-containing lipoproteins enriched with triglycerides and is thus a promising target for the treatment of hyperlipidemia. Yet, the development of MTP inhibitors to lower plasma lipid concentrations has been hindered by adverse effects on hepatic steatosis. A study recently published in Nature Medicine identifies microRNA-30c (miR-30c) as a potent repressor of MTP that controls plasma apoB-containing lipoprotein levels, in addition to decreasing hepatic lipid synthesis through direct targeting of lysophosphatidylglycerol acyltransferase 1 (LPGAT1). These findings identify miR-30c as a novel therapeutic target that coordinately reduces lipid biosynthesis and lipoprotein secretion to suppress circulating apoB lipoproteins, while sparing the liver from steatosis.

RWTH Aachen University, Diss., 2017; 88 pp. (2017). = RWTH Aachen University, Diss., 2017, Two of the most important process parameters for aerobic cultivations of microorganisms and cells are the oxygen consumption and oxygen content within the cultivation broth. By means of the consumption, mass balances and stoichiometries can be calculated. On the basis of the oxygen content, an oxygen limitation can be identified doubtlessly. Additionally, the propagation of these parameters can be seen as a characteristic physiological fingerprint of an aerobic culture. Both parameters enable a sound and valid process monitoring and characterization. For these reasons, respective sensors and analytics are commonly applied in fermenter scale. However, the first steps of bioprocess development are generally performed in small scale experiments within shake flasks or microtiter plates. This is due to the ability to use smaller reaction volumes while performing a greater number of parallel experiments, which saves time, resources and consequently lowers the overall costs per experiment. Unfortunately, due to the small dimensions, respective sensors and analytics are not easily transferable from fermenter to small scale and the current solutions are not completely satisfying. To augment the process knowledge already in early process development, it is of general interest to develop new tools and improve existing technologies for small-scale bioreactors. Within this work, three different novel systems for online monitoring of aerobic cultures in small scale fermentations have been developed, introduced and characterized.Firstly, a novel shake flask based system for measuring the dissolved oxygen tension (DOT) is introduced. By using dispersed oxygen-sensitive nanoparticles, an easy to use, robust and reliable DOT measurement system was developed, which is applicable for almost all cultivation conditions in shake flasks and for both, soluble complex and synthetic media. Reliable DOT measurements were achieved using oxygen-sensitive nanoparticles added to the cultivation broth at a concentration of 0.1 g L 1. The biocompatibility of the dispersed oxygen-sensitive nanoparticles was demonstrated for H. polymorpha, X. campestris and E. coli by means of RAMOS cultivations. Furthermore, the importance of system calibration under cultivation conditions was demonstrated. Besides that, the ability of the dispersed oxygen-sensitive nanoparticles to withstand autoclaving is an additional advantage for sterile cultivations. This measurement system turned out to be a valuable alternative for existing DOT measurement systems in shake flask cultivations, especially at low filling volumes and high shaking frequencies. In combination with RAMOS data it was possible to determine kLa values online during cultivations of K. lactis, E. coli and H. polymorpha. The determined kLa values were investigated with respect to their corresponding uncertainties caused by systematic errors in OTR, DOT, L_(O_2 ) and p_(O_2)^gas estimations and sensor inertia. All kLa values were in good agreement with an empirical correlation based on the medium osmolality from the literature. Secondly, this oxygen-sensitive nanoparticle-based technique was transferred to the microtiter plate scale and combined with the established BioLector technology. The respective optical fiber of the oxygen analytic was integrated into the BioLector system and displaced via the BioLector movement axes underneath the microtiter plate. Since this system does not require the presence of optodes at the bottom of each well, the measurement system can be applied to various microtiter plates (MTPs). Finally, the cost constraints associated with MTPs possessing immobilized sensor spots (optodes) can be overcome using this technology. Additionally, the costs per cultivation are significantly reduced and the system provides a serious alternative to established devices. Furthermore, kLa values for the applied cultivation conditions were estimated and used to calculate the oxygen transfer rates (OTRs) in MTPs based on the DOT. OTRs calculated for the MTPs and OTRs determined in shake flasks via the RAMOS technology agreed very well with each other.Lastly, a microtiter plate-based µRAMOS system was developed to measuring the oxygen transfer rate (OTR) in every individual well of a 48-well microtiter plate. On the basis of a first four-well prototype a second enhanced prototype covering the whole 48-well microtiter plate was implemented. The necessary valve and sensor technology was thereby successfully integrated in a microfluidic microtiter plate cover. To compromise on sufficient measurement rates and an economically acceptable number of oxygen instruments an 8x48 optical multiplexer (MUX) was developed and utilized. By means of four exemplary RAMOS and µRAMOS cultivations, both systems delivered the same OTR signal quality. The cultivation throughput could be increased 6-fold compared to the established eight shake flask RAMOS system. Additionally, the media consumption was reduced 12.5-fold per cultivation from a typical filling volume of 10 mL in the shake flask to 800 µL in the microtiter plate. In summary, three novel measurement systems were successfully developed, built and characterized. These systems offer a sound determination of the dissolved oxygen tension and oxygen transfer rate respectively and turned out to be predestined small scale analytics for process development and characterization purposes., Published by Aachen

Metatarsophalangeal (MTP) joint osteoarthritis (OA), also known as hallux rigidus (HR), is the most common degenerative arthropathy of the foot and is often the result of trauma. There are multiple methods of addressing the patient’s pain and limited function. Arthrodesis is the gold standard to manage severe MTP arthritis with a highly significant union rate. With various techniques of arthrodesis available, ranging from cannulated screw fixation, Kirschner wires, as well as plate and screw fixation, the orthopedic surgeon has multiple modalities to address this ailment; however, when these fail due to infection, the armament is limited. Through the idea of articulating antibiotic spacers in other regions of the body such as the knee and hip, we present a novel technique to the creation of an antibiotic spacer in the setting of a failed infected MTP arthrodesis.

Fat is delivered to tissues by apoB-containing lipoproteins synthesized in the liver and intestine with the help of an intracellular chaperone, microsomal triglyceride transfer protein (MTP). Leptin, a hormone secreted by adipose tissue, acts in the brain and on peripheral tissues to regulate fat storage and metabolism. Our aim was to identify the role of leptin signaling in MTP regulation and lipid absorption using several mouse models deficient in leptin receptor (LEPR) signaling and downstream effectors. Mice with spontaneous LEPR B mutations or targeted ablation of LEPR B in proopiomelanocortin (POMC) or agouti gene related peptide (AGRP) expressing cells had increased triglyceride in plasma, liver, and intestine. Furthermore, melanocortin 4 receptor (MC4R) knockout mice expressed a similar triglyceride phenotype, suggesting that leptin might regulate intestinal MTP expression through the melanocortin pathway. Mechanistic studies revealed that the accumulation of triglyceride in the intestine might be secondary to decreased expression of MTP and lipid absorption in these mice. Surgical and chemical blockade of vagal efferent outflow to the intestine in wild-type mice failed to alter the triglyceride phenotype, demonstrating that central neural control mechanisms were likely not involved in the observed regulation of intestinal MTP. Instead, we found that enterocytes express LEPR, POMC, AGRP, and MC4R. We propose that a peripheral, local gut signaling mechanism involving LEPR B and MC4R regulates intestinal MTP and controls intestinal lipid absorption.

Die vorliegende Arbeit untersucht die Vorteilhaftigkeit modularer verfahrenstechnischer Produktionsanlagen. Der Schwerpunkt liegt dabei auf den Engineering-Aktivitäten im Gewerk Automatisierungstechnik. Insbesondere wird beleuchtet, wie groß der Anteil des zeitkritischen Anlagen-Engineerings ist, der durch die Verwendung von Modulen eingespart werden kann. Die Ergebnisse der Untersuchung beschreiben die Abhängigkeiten von den automatisierungstechnischen Auslegungsmöglichkeiten der Produktionsmodule und der Produktionsinfrastruktur. Letztendlich wird dargestellt, wie eine Verringerung des zeitkritischen Gesamtanlagen-Engineering auf 50% erreicht werden kann.

Background/Aims: Apolipoprotein E (apoE)-deficient mice develop hepatic steatosis and secrete reduced levels of VLDL-TG. Methods and results: We examined the effects of apoE-deficiency on intracellular lipid homeostasis and secretion of triglycerides (TG). We show that intracellular TG turnover and activities of diacylglycerol acyltransferase (DGAT) and microsomal triglyceride transfer protein (MTP) are similar in Apoe(-/-) and wild type mice. In addition, apoB synthesis was not decreased in Apoe(-/-) cells. Thus, the accumulation of lipid in these cells is not attributable to perturbed TG turnover, apoB synthesis, and the activities of DGAT and MTP. Inhibition of MTP had a more profound impact on the secretion of VLDL-TG from wild type hepatocytes than Apoe(-/-) hepatocytes, indicating that MTP was more limiting for the production of VLDL-TG from wild type cells. In marked contrast to the MTP-deficient model of fatty liver, electron microscopy of lipid-stained liver sections of Apoe(-/-) mice revealed an accumulation of lipid in numerous small, putative ER-derived vesicles and in the cytosol. No abnormalities were observed in the Golgi of Apoe(-/-) mice. Conclusions: These results suggest that the removal of lipids from the early or intermediary compartments of the secretory pathway of hepatocytes is impaired in Apoe(-/-) mice. (C) 2004 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Les anomalies du métabolisme des lipoprotéines riches en triglycérides (LRTG), les chylomicrons et les VLDL exposent à des hypocholestérolémies lors d'un défaut de sécrétion et à des hypertriglycéridémies (HTG) majeures entraînant un risque athéromateux et de pancréatites aigües lors de l'altération de leur clairance. Nous avons diagnostiqué des patients présentant un défaut génétique de sécrétion des LRTG au décours de maladie de rétention des chylomicrons, d'abetalipoprotéinémie et d'hypobetalipoprotéinémie homozygote, causées respectivement par des mutations sur les gènes SAR1B, MTTP et APOB. Nous avons étudié le phénotype des 158 patients publiés avec mutation délétère et mis en évidence des différences portant principalement sur la stéatose hépatique, l'insulinorésistance et l'obésité. Nous avons également mis au point une méthode d'évaluation de l'activité post héparinique de la lipoprotéine lipase (LPL) par mesure de la lipolyse des triglycérides des VLDL in vitro, permettant l'exploration phénotypique des patients présentant une HTG sévère. Nous avons mis en évidence des activités LPL augmentées chez des patients présentant pourtant des antécédents d'HTG sévère et des déficits chez des patients ne présentant pas de mutation identifiable du gène LPL, laissant supposer l'existence de facteurs additionnels modulant l'expression ou l'activité de la LPL. Enfin des interrelations des multiples gènes impliqués dans le métabolisme des triglycérides modulent le phénotype. Elles soulèvent l'intérêt de l'exploration simultanée des principaux gènes impliqués dans les dyslipidémies, telle qu'elle sera effectuée par NGS, pour une meilleure compréhension de leur physiopathologie, Abnormal metabolism of triglyceride-rich lipoproteins (LRTG), chylomicrons and VLDL, can result in hypocholesterolemia in case of impaired secretion, or severe hypertriglyceridemia (HTG) and increased risk of atheroma and acute pancreatitis if clearance is affected. We explored patients suffering from genetic defect in the LRTG secretion (chylomicron retention disease, abetalipoproteinemia and homozygous hypobetalipoproteinemia) and identified mutations on respectively SAR1B, and MTTP and APOB gene. Then, we analysed the phenotype of 158 previously published patients with deleterious mutation (i.e. reported cases added to our cohort) and were able to highlight some specific differences like hepatic steatosis, insulin resistance and obesity. Furthermore we developed an assay to evaluate the lipoprotein lipase (LPL) functionality by measuring the triglyceride-VLDL lipolysis in vitro, and provide a reliable phenotypic exploration for patients with past history of severe hypertriglyceridemia. We found an increased LPL activity in some patients with severe hypertriglyceridemia but conversely showed deficits in other patients free from mutation on LPL gene. These results lead to hypothesize that additional factors might contribute to modulate the expression or the activity of LPL. Finally multiple genes of triglycerides metabolism interact together to additionally modulate phenotype. Of high interest is therefore the simultaneous exploration of the key genes involved in dyslipidemia, as provided by the new generation sequencing (NGS), for better understanding of all pathophysiological mechanisms

Hypercholesterolemia represents a major risk factor of cardiovascular disease. A number of experimental models is used for study of hypercholesterolemia pathogenesis and therapy. This thesis concentrates on characterization of one of these models. Prague hereditary hypercholesterolemic (PHHC) rat is such a suitable model for study of hypercholesterolemia. Although the majority of plasma cholesterol is transported by high density lipoprotein in PHHC rat fed standard diet, PHHC rat fed cholesterol diet develops hypercholesterolemia comparable to that of humans. The advantage of this model is that hypercholesterolemia develops without the need for addition of bile acids or other hepatotoxic substances to the diet. The hypercholesterolemia of PHHC rat is caused by slowed down catabolism of cholesterol-rich very low density lipoprotein (VLDL). These cholesterol-rich particles are synthesized in the liver. We found out that PHHC rat fed 1% cholesterol diet accumulates cholesteryl esters (CE) in the liver and also in the VLDL. Acyl-CoA:cholesterol acyltransferase (ACAT) and microsomal triglyceride transfer protein (MTP) may participate in the increased incorporation CE into VLDL. We found out no difference in ACAT and MTP activities in the liver between PHHC rats and control animals. Neither ACAT activity...

International audience; The membrane glycoprotein CD36 binds nanomolar concentrations of long chain fatty acids (LCFA) and is highly expressed on the luminal surface of enterocytes. CD36 deficiency reduces chylomicron production through unknown mechanisms. In this report, we provide novel insights into some of the underlying mechanisms. Our in vivo data demonstrate that CD36 gene deletion in mice does not affect LCFA uptake and subsequent esterification into triglycerides by the intestinal mucosa exposed to the micellar LCFA concentrations prevailing in the intestine. In rodents, the CD36 protein disappears early from the luminal side of intestinal villi during the postprandial period, but only when the diet contains lipids. This drop is significant 1 h after a lipid supply and associates with ubiquitination of CD36. Using CHO cells expressing CD36, it is shown that the digestion products LCFA and diglycerides trigger CD36 ubiquitination. In vivo treatment with the proteasome inhibitor MG132 prevents the lipid-mediated degradation of CD36. In vivo and ex vivo, CD36 is shown to be required for lipid activation of ERK1/2, which associates with an increase of the key chylomicron synthesis proteins, apolipoprotein B48 and microsomal triglyceride transfer protein. Therefore, intestinal CD36, possibly through ERK1/2-mediated signaling, is involved in the adaptation of enterocyte metabolism to the postprandial lipid challenge by promoting the production of large triglyceride-rich lipoproteins that are rapidly cleared in the blood. This suggests that CD36 may be a therapeutic target for reducing the postprandial hypertriglyceridemia and associated cardiovascular risks.

Se propone una estrategia de inversión para el “pequeño inversionista ” en los mercados de valores internacionales con los instrumentos denominados ETF. Se trata de un trabajo empírico que utiliza la Teoría Moderna del Crecimiento de la Economía, la Teoría Moderna de Construcción de Portafolios y la Teoría de distribución de probabilidades. De esta forma conjuga a la economía en su sentido puro, a las finanzas en su desarrollo teórico práctico y a la estadística con sus desarrollos metodológicos, para construir una estrategia que aporta al inversionista suficiente información para realizar inversiones alrededor del mundo, obteniendo atractivos rendimientos y al mismo tiempo minimizando el riesgo. Los resultados de la aplicación de la estrategia, muestran que los rendimientos son mayores a los obtenidos por activos de renta fija y fondos mutuos de inversión de renta variable focalizados en activos del mercado internacional. _______________ ABSTRACT: We are proposing an investment strategy for "small investors " in the international stock market using instruments denominated ETF. It is an empirical work that uses the Modern Theory of Economic Growth, the Modern Theory of Portfolio and the Theory of probability distribution. We combine economy in its pure sense, practical and theoretical developments in finance and methodological developments in statistics, to build a strategy which provides sufficient information to investors in order to make international investment, obtain good returns and minimize risk. The results of the implementation of this strategy, show that the returns are higher than those obtained by fixed assets and mutual funds centered on the international market.

Thus far, liver, intestine, heart, and placenta have been shown to secrete apolipoprotein (apo) B-containing lipoproteins. In the present study, we first investigated lipoproteins in human follicular fluid (FF), surrounding developing oocytes within the ovary, as well as in corresponding plasma samples (n = 12). HDL cholesterol within FF correlated well with plasma HDL cholesterol (r = 0.80, P