Effect of Revascularization on Intramuscular Vascular Endothelial Growth Factor Levels in Peripheral Arterial Disease

Vascular endothelial growth factor (VEGF) is a potent driver of angiogenesis, which may help to relieve ischemia in peripheral arterial disease (PAD). We aimed to investigate the role of intramuscular VEGF in ischemic and non-ischemic skeletal muscle in PAD patients before and after surgical or endovascular revascularization and different stages of PAD. Biopsies of the gastrocnemius and vastus muscles from twenty PAD patients with stenosis or occlusion of the superficial femoral artery were obtained both during revascularization and 8 weeks postoperatively. The gastrocnemius muscle was considered ischemic, while vastus muscle biopsies served as intraindividual controls. The levels of vascular endothelial growth factor in muscle lysates were then determined by ELISA. Preoperative VEGF levels were significantly higher in ischemic muscles compared to the controls (98.07 ± 61.96 pg/mL vs. 55.50 ± 27.33 pg/mL, p = 0.004). Postoperative values decreased significantly (p = 0.010) to 54.83 ± 49.60 pg/mL in gastrocnemius biopsies. No significant change was observed in vastus muscle biopsies, with mean postoperative VEGF values found at 54.16 ± 40.66 pg/mL. Since all patients still had indications for revascularization, impairment of angiogenesis mechanisms can be assumed. More research about angiogenesis in PAD is needed with the ultimate goal to improve conservative treatment

A time-resolved proteomic and prognostic map of COVID-19

COVID-19 is highly variable in its clinical presentation, ranging from asymptomatic infection to severe organ damage and death. We characterized the time-dependent progression of the disease in 139 COVID-19 inpatients by measuring 86 accredited diagnostic parameters, such as blood cell counts and enzyme activities, as well as untargeted plasma proteomes at 687 sampling points. We report an initial spike in a systemic inflammatory response, which is gradually alleviated and followed by a protein signature indicative of tissue repair, metabolic reconstitution, and immunomodulation. We identify prognostic marker signatures for devising risk-adapted treatment strategies and use machine learning to classify therapeutic needs. We show that the machine learning models based on the proteome are transferable to an independent cohort. Our study presents a map linking routinely used clinical diagnostic parameters to plasma proteomes and their dynamics in an infectious disease

Clinical and virological characteristics of hospitalised COVID-19 patients in a German tertiary care centre during the first wave of the SARS-CoV-2 pandemic: a prospective observational study

Purpose: Adequate patient allocation is pivotal for optimal resource management in strained healthcare systems, and requires detailed knowledge of clinical and virological disease trajectories. The purpose of this work was to identify risk factors associated with need for invasive mechanical ventilation (IMV), to analyse viral kinetics in patients with and without IMV and to provide a comprehensive description of clinical course. Methods: A cohort of 168 hospitalised adult COVID-19 patients enrolled in a prospective observational study at a large European tertiary care centre was analysed. Results: Forty-four per cent (71/161) of patients required invasive mechanical ventilation (IMV). Shorter duration of symptoms before admission (aOR 1.22 per day less, 95% CI 1.10-1.37, p < 0.01) and history of hypertension (aOR 5.55, 95% CI 2.00-16.82, p < 0.01) were associated with need for IMV. Patients on IMV had higher maximal concentrations, slower decline rates, and longer shedding of SARS-CoV-2 than non-IMV patients (33 days, IQR 26-46.75, vs 18 days, IQR 16-46.75, respectively, p < 0.01). Median duration of hospitalisation was 9 days (IQR 6-15.5) for non-IMV and 49.5 days (IQR 36.8-82.5) for IMV patients. Conclusions: Our results indicate a short duration of symptoms before admission as a risk factor for severe disease that merits further investigation and different viral load kinetics in severely affected patients. Median duration of hospitalisation of IMV patients was longer than described for acute respiratory distress syndrome unrelated to COVID-19

Effects of therapy with the non-steroidal androgen receptor modulator Ligandrol on the skeletal muscle in the ovariectomized rat model

Sarkopenie ist die Bezeichnung für ein Syndrom, welches sich durch einen fortschreitenden Verlust von Skelettmuskelmasse und -kraft auszeichnet und mit einem ungünstigen Outcome bis hin zum Tod vergesellschaftet ist. Diese generalisierte Form der Skelettmuskelatrophie geht auch dem Krankheitsbild der Osteoporose voraus und lässt sich häufig auf ähnliche pathophysiologische Faktoren wie z. B. latente chronisch entzündliche Prozesse und einen Mangel an anabolen Hormonen wie Testosteron oder Östrogen zurückführen. Auch als Konsequenz der weltweit steigenden Lebenserwartung des Menschen weisen Sarkopenie und Osteoporose als Folge der physiologischen Menopause steigende Inzidenzen auf. Die Substitution von Androgenen ist der wohl naheliegendste Therapieansatz, weist allerdings ein nicht unerhebliches Spektrum an Nebenwirkungen auf. Die Entwicklung der selektiven Androgenrezeptormodulatoren (SARMs) ist das Resultat einer Suche nach einem Androgenrezeptor-Stimulans, welches oral bioverfügbar und gewebeselektiver ist als z. B. Testosteron. Der Androgenrezeptor ist ein ubiquitär im Körper exprimierter Rezeptor, welcher als Transkriptionsfaktor fungiert und durch androgene Hormone aber auch Substanzen wie die nicht-steroidalen selektiven Androgenrezeptormodulatoren (SARMs) angesteuert werden kann und resultierend in verschiedenen Geweben einen unterschiedlich stark ausgeprägten anabolen Stimulus induziert. Ligandrol oder LGD 4033 ist ein oraler nichtsteroidaler SARM mit hoher Affinität zum AR und guter Verträglichkeit. Ziel dieser Studie ist es, die Auswirkungen einer Therapie mit Ligandrol auf die Skelettmuskulatur im ovarektomierten Rattenmodell für postmenopausale Bedingungen zu untersuchen. Zu diesem Zweck wurde ein Versuchskollektiv von 75 weiblichen Sprague-Dawley Ratten im Alter von drei Monaten in fünf Versuchsgruppen zu je 15 Tieren eingeteilt. Eine Kontrollgruppe wurde unversehrt belassen (Non OVX), eine Gruppe wurde ovarektomiert und nicht behandelt (OVX) und drei Gruppen wurden ovarektomiert und neun Wochen nach der Ovarektomie mit Ligandrol in einer Dosierung von 0,04 mg/kg KG (OVX niedrig), 0,4 mg/kg KG (OVX mittel), und 4 mg/kg KG (OVX hoch) für 35 Tage mittels Applikation über das Futter behandelt. Nach einer Versuchsdauer von insgesamt 14 Wochen erfolgte die Tötung und Entnahme der Muskeln M. gastrocnemius, M. soleus und M. longissimus. Diese wurden auf Kapillarisierung, Muskelfaserquerschnittsfläche und Zellzusammensetzung untersucht. Zudem entnommen wurden die Uteri und Blutproben zur Analyse der Serum-Creatinkinase. Die Behandlung mit Ligandrol bewirkte im gegebenen Versuchszeitraum eine zum Körpergewicht proportionale Steigerung der Kapillarisierung in allen untersuchten Muskelgruppen. Es war eine Steigerung des Muskelfaserquerschnitts im M. longissimus in der OVX hoch-Gruppe sowie im M. soleus in den Gruppen OVX und OVX mittel zu verzeichnen. Gleichzeitig trat unter der Behandlung mit 4,0 mg/kg KG Ligandrol eine signifikante Hypertrophie der Uteri auf, weswegen davon auszugehen ist, dass die Gewebeselektivität des Medikaments eine Dosisabhängigkeit aufweist. Das durchschnittliche Gewicht des M. gastrocnemius steigerte sich nach der Ovarektomie sowie dosisabhängig im Laufe der Therapie. Ligandrol kann also eine Steigerung der Muskelfaserquerschnittsfläche bewirken und verbessert über eine Steigerung der Kapillarisierung die muskuläre Durchblutung. Zusammenfassend lässt sich sagen, dass Ligandrol sich dosisabhängig positiv auf die Muskulatur im ovarektomierten Rattenmodell auswirken kann. Unter einer Behandlung mit 4 mg/kg KG Ligandrol täglich kann die Muskelfaserquerschnittsfläche vergrößert und bereits mit 0,04 mg/kg KG die muskuläre Durchblutung über eine Steigerung der Kapillarisierung verbessert werden. Weitere Studien müssen allerdings diejenige Dosis ermitteln, ab der eine Gewebeselektivität nicht mehr besteht und aufzeigen, ob bei einer unkritischen Dosierung gegebenenfalls über einen längeren Versuchszeitraum eine Steigerung der Muskelfaserquerschnittsfläche, Knochenstruktur oder eine Verbesserung der Kapillarisierung bewirkt werden kann. Insbesondere muss untersucht werden, ob die Zunahme des Muskelgewichtes tatsächlich durch eine Hypertrophie der Muskelfasern zustande kommt und ob diese und die nachweislich gesteigerte Kapillarisierung auch mit einer verbesserten muskulären Funktion einhergehen.Sarcopenia denotes a syndrome that is characterized by a progressive loss of skeletal muscle mass and strength and is associated with unfavourable outcomes including death. This generalized form of skeletal muscle atrophy also precedes the clinical presentation of osteoporosis and can often be attributed to similar pathophysiological factors such as latent chronic inflammatory processes and a lack of anabolic hormones such as testosterone or oestrogen. Also, because of a worldwide increase of human life expectancy, sarcopenia and osteoporosis show increasing incidences as a result of physiological menopause. The substitution of androgens is probably the most obvious therapeutic approach but goes along with a whole spectrum of possible severe side effects. The development of selective androgen receptor modulators (SARMs) is the result of a search for an androgen receptor stimulant that is orally bioavailable and more tissue-selective than, for example, testosterone. The androgen receptor is ubiquitously expressed in the human body and acts as a transcription factor which can be controlled by androgenic hormones but also substances such as the SARMs and as a result induces a different degree of anabolic stimuli in different tissues. Ligandrol or LGD 4033 is an orally applicable nonsteroidal SARM with high affinity to the androgen receptor and a good overall tolerability. The aim of this study is to investigate the effects of a therapy with Ligandrol on skeletal muscle in the ovariectomized rat model for postmenopausal conditions. For this purpose, an experimental group of 75 female Sprague-Dawley rats at the age of three months was divided into five experimental groups of 15 animals each. One control group was left intact (Non OVX), one group was ovariectomized and untreated (OVX) and three groups were ovariectomized and treated nine weeks after ovariectomy with Ligandrol at a dosage of 0.04 mg/kg bodyweight (OVX low), 0.4 mg/kg bodyweight (OVX medium), and 4 mg/kg bodyweight (OVX high) for 35 days administered with the feed. After a trial period of a total of 14 weeks, the collective was killed and the muscles M. gastrocnemius, M. soleus and M. longissimus were removed. These were examined for capillarization, muscle fibre cross-sectional area and cell composition. In addition, the uterus and blood samples were taken for the analysis of serum creatine kinase. Treatment with Ligandrol resulted in an increase in capillarization proportional to body weight in all muscle groups studied during the given trial period. There was an increase in the muscle fibre cross-section in the longissimus muscle in the OVX high group and in the M. soleus in the OVX and OVX medium groups. At the same time, significant hypertrophy of the uterus occurred during treatment with 4.0 mg/kg bodyweight Ligandrol, which is why it can be assumed that the tissue selectivity of the drug has a dose dependence. The average weight of the gastrocnemius muscle increased after the ovariectomy as well as dose-dependent during therapy. Ligandrol can therefore cause an increase in the muscle fibre cross-sectional area and improves the muscular blood circulation by increasing capillarization. In summary, Ligandrol can have a dose-dependent positive effect on the muscles in the ovariectomized rat model. With a dose as low as 0.04 mg/kg bodyweight the muscular blood circulation can be improved by increasing capillarization. Under treatment with 4 mg/kg bodyweight Ligandrol daily, the muscle fibre cross-sectional area can be increased. However, further studies must determine the dose above which tissue selectivity no longer exists and show whether an increase in muscle fibre cross-sectional area, bone structure or an improvement in capillarization can be achieved with an uncritical dosage over a longer experimental period. Also, it must be investigated whether the increase in muscle weight is really caused by hypertrophy of the muscle fibres and whether both this and the increased capillarization are associated with improved muscular function.2022-09-2

Effect of Revascularization on Intramuscular Vascular Endothelial Growth Factor Levels in Peripheral Arterial Disease

Vascular endothelial growth factor (VEGF) is a potent driver of angiogenesis, which may help to relieve ischemia in peripheral arterial disease (PAD). We aimed to investigate the role of intramuscular VEGF in ischemic and non-ischemic skeletal muscle in PAD patients before and after surgical or endovascular revascularization and different stages of PAD. Biopsies of the gastrocnemius and vastus muscles from twenty PAD patients with stenosis or occlusion of the superficial femoral artery were obtained both during revascularization and 8 weeks postoperatively. The gastrocnemius muscle was considered ischemic, while vastus muscle biopsies served as intraindividual controls. The levels of vascular endothelial growth factor in muscle lysates were then determined by ELISA. Preoperative VEGF levels were significantly higher in ischemic muscles compared to the controls (98.07 ± 61.96 pg/mL vs. 55.50 ± 27.33 pg/mL, p = 0.004). Postoperative values decreased significantly (p = 0.010) to 54.83 ± 49.60 pg/mL in gastrocnemius biopsies. No significant change was observed in vastus muscle biopsies, with mean postoperative VEGF values found at 54.16 ± 40.66 pg/mL. Since all patients still had indications for revascularization, impairment of angiogenesis mechanisms can be assumed. More research about angiogenesis in PAD is needed with the ultimate goal to improve conservative treatment.info:eu-repo/semantics/publishe

In Situ Monitoring of Scale Effects on Phase Selection and Plasmonic Shifts during the Growth of AgCu Alloy Nanostructures for Anticounterfeiting Applications

Tailoring of plasmon resonances is essential for applications in anti-counterfeiting. This is readily achieved by tuning the composition of alloyed metal clusters; in the most simple case binary alloys. Yet, one challenge is the correlation of cluster morphology and composition with the changing optoelectronic properties. Hitherto, the early stages of metal alloy nanocluster formation in immiscible binary systems like silver and copper has been accessible by molecular dynamics simulations and transmission electron microscopy. Here, we investigate in real-time the formation of supported silver, copper and silver-copper-alloy nanoclusters during sputter deposition on poly(methyl methacrylate) by combining in situ surface sensitive X-ray scattering with optical spectroscopy. While following the transient growth morphologies, we quantify the early stages of phase separation at the nanoscale, follow the shifts of surface plasmon resonances and quantify the growth kinetics of the nanogranular layers at different thresholds. We are able to extract the influence of scaling effects on the nucleation and phase selection. The internal structure of the alloy cluster shows a copper-rich core/silver-rich shell structure, since the copper core yields a lower mobility and higher crystallization tendency than the silver fraction. We compare our results to molecular dynamics simulation and transmission electron microscopy data. This demonstrates a route to tailor accurately the plasmon resonances of nanosized, polymer supported clusters which is a crucial prerequisite for anti-counterfeiting

Clinical and virological characteristics of hospitalised COVID-19 patients in a German tertiary care centre during the first wave of the SARS-CoV-2 pandemic: a prospective observational study

Purpose!#!Adequate patient allocation is pivotal for optimal resource management in strained healthcare systems, and requires detailed knowledge of clinical and virological disease trajectories. The purpose of this work was to identify risk factors associated with need for invasive mechanical ventilation (IMV), to analyse viral kinetics in patients with and without IMV and to provide a comprehensive description of clinical course.!##!Methods!#!A cohort of 168 hospitalised adult COVID-19 patients enrolled in a prospective observational study at a large European tertiary care centre was analysed.!##!Results!#!Forty-four per cent (71/161) of patients required invasive mechanical ventilation (IMV). Shorter duration of symptoms before admission (aOR 1.22 per day less, 95% CI 1.10-1.37, p &amp;lt; 0.01) and history of hypertension (aOR 5.55, 95% CI 2.00-16.82, p &amp;lt; 0.01) were associated with need for IMV. Patients on IMV had higher maximal concentrations, slower decline rates, and longer shedding of SARS-CoV-2 than non-IMV patients (33 days, IQR 26-46.75, vs 18 days, IQR 16-46.75, respectively, p &amp;lt; 0.01). Median duration of hospitalisation was 9 days (IQR 6-15.5) for non-IMV and 49.5 days (IQR 36.8-82.5) for IMV patients.!##!Conclusions!#!Our results indicate a short duration of symptoms before admission as a risk factor for severe disease that merits further investigation and different viral load kinetics in severely affected patients. Median duration of hospitalisation of IMV patients was longer than described for acute respiratory distress syndrome unrelated to COVID-19

Somatic mosaicism and common genetic variation contribute to the risk of very-early-onset inflammatory bowel disease

Very-early-onset inflammatory bowel disease (VEO-IBD) is a heterogeneous phenotype associated with a spectrum of rare Mendelian disorders. Here, we perform whole-exome-sequencing and genome-wide genotyping in 145 patients (median age-at-diagnosis of 3.5 years), in whom no Mendelian disorders were clinically suspected. In five patients we detect a primary immunodeficiency or enteropathy, with clinical consequences (XIAP, CYBA, SH2D1A, PCSK1). We also present a case study of a VEO-IBD patient with a mosaic de novo, pathogenic allele in CYBB. The mutation is present in ~70% of phagocytes and sufficient to result in defective bacterial handling but not life-threatening infections. Finally, we show that VEO-IBD patients have, on average, higher IBD polygenic risk scores than population controls (99 patients and 18,780 controls; P &lt; 4 × 10-10), and replicate this finding in an independent cohort of VEO-IBD cases and controls (117 patients and 2,603 controls; P &lt; 5 × 10-10). This discovery indicates that a polygenic component operates in VEO-IBD pathogenesis