Grayanotoxin I Intoxication in Pet Pigs

Contaminated honey is a common cause of grayanotoxin intoxication in humans. Intoxication of animals, especially cattle, is usually due to ingestion of plants of the Ericaceae family, such as Rhododendron. Here, we report the ingestion of Pieris japonica as the cause of grayanotoxin I intoxication in 2 miniature pigs that were kept as pets. The pigs showed sudden onset of pale oral mucosa, tachycardia, tachypnea, hypersalivation, tremor, and ataxia that progressed to lateral recumbency. The pathological examination of one pig revealed no specific indications for intoxication except for the finding of plant material of Pieris japonica in the intestine. Grayanotoxin I was identified in the ingested plant, gastric content, blood, liver, bile, kidney, urine, lung, and skeletal muscle via HPLC-MS/MS. Grayanotoxin I should be considered as a differential etiological diagnosis in pigs with unspecific signs and discovery of ingested plant material as the only indication in the pathologic examination

Core-Multishell-Nanocarrier für den topischen Wirkstofftransport in einem Psoriasis Mausmodell

Psoriasis is a common, chronic, multifactorial, human skin disease, characterized by well-demarcated, raised, erythematous plaques covered with silvery scales. It is incurable and often requires long period therapy with immunomodulatory drugs, which can lead to side effects, especially when administered systemically in more severe cases. Nanocarrier are engineered particles of a size between 1 nm and 100 nm at least in one dimension. For skin, nanocarrier are designed to increase the delivery of drugs or genes through the skin barrier, target the drug to a specific layer, or prevent systemic distribution and thereby negative effects in distant organs. In paper 1 of this thesis, core-multishell nanocarrier were investigated topically on mouse skin in vivo. This nanocarrier had been designed like a uni-molecular micelle with a hydrophilic core, an inner lipophilic shell, and an outer hydrophilic shell to make the particle water-soluble and provide space for drugs of different lipophilicity to be loaded into the core or the inner shell. Originally, they had been thought to penetrate through the skin and release their cargo at its site of action. Literature already had stated that several nanocarrier or nanoparticles do not penetrate into skin, whereas others do and even others penetrate only through a disrupted barrier. The core-multishell nanocarrier used here did not penetrate into viable layers of intact or inflamed skin using the imiquimod-induced psoriasis model in BALB/c mice. Instead, they accumulated in the stratum corneum. Previous in vitro data had shown core-multishell nanocarrier penetration into tape stripped human skin ex vivo. The accumulation in the stratum corneum could possibly be used as a depot for a retarded and prolonged release of drugs. Parallel to the penetration study no adverse effects were observed locally or systemically. This is in concordance with literature also stating no negative effects even after repeated subcutaneous injection of this carrier. The topical application of nile red loaded core-multishell nanocarrier revealed superior cargo delivery into the viable epidermis compared to a nile red cream. Further elucidation of the mechanism by which the core-multishell nanocarrier enhances the penetration of nile red, and proof of concept for real drugs are needed. However, the core-multishell nanocarrier remains as a promising candidate for further development for therapy of skin diseases including testing of a therapeutic surplus value of drug loaded nanocarrier compared to commercial topical drug formulations. No tacrolimus-specific anti-inflammatory effects could be shown in the imiquimod-induced psoriasis-like dermatitis model using BALB/c mice, despite penetration of the drug into the dermis. Further research is needed to elucidate the reason for the lack of that efficacy and the conflict to literature and the dependence of tacrolimus efficacy in this model on mouse strains. However, this model was not applicable to evaluate therapeutic superiority of core-multishell nanocarrier for tacrolimus delivery compared to tacrolimus ointment. The importance of choosing an appropriate model for the specific question, using multiple objective readout parameters to avoid over-interpretation of small variations, and testing against all needed control groups, including a vehicle control, in addition to untreated controls, is highlighted.Psoriasis ist eine häufige, chronische, multifaktorielle Hauterkrankung des Menschen, welche durch scharf begrenzte, erhabene, erythematöse Plaques, die mit silbrigen Schuppen bedeckt sind, charakterisiert ist. Psoriasis ist unheilbar und oft ist eine Langzeittherapie mit immunmodulatorischen Medikamenten nötig, welche vor allem in schweren Fällen bei systemischer Applikation zu unerwünschten Nebenwirkungen führen können. Nanocarrier sind synthetische Partikel von einer Größe zwischen 1 nm und 100 nm in mindestens einer Dimension. Für die Anwendung auf der Haut wurden Nanocarrier entwickelt um, Wirkstoffe oder Gene vermehrt durch die Hautbarriere zu transportieren, bestimmte Schichten anzusteuern oder eine systemische Verteilung des Wirkstoffes, und dabei Nebenwirkungen in Organen, zu verhindern. In der ersten Publikation aus dieser Dissertation wurden Core-Multishell Nanocarrier in vivo topisch auf Maushaut getestet. Dieser Nanocarrier wurde in Analogie zu einer ein-molekularen Mizelle mit einem hydrophilen Kern, einer lipophilen inneren Schale und einer hydrophilen äußeren Schale entwickelt, um den Partikel wasserlöslich zu machen und Wirkstoffe verschiedener Lipophilität im Kern und der inneren Schale transportieren zu können. Ursprünglich sollten die Nanocarrier in die Haut eindringen und ihre Ladung an dessen Wirkungsort freigeben. In der Literatur ist bereits zu finden, dass verschiedene Nanocarrier und Nanopartikel nicht in Haut eindringen, wobei andere dieses tun und wieder andere nur in barrieregestörte Haut eindringen. Die hier genutzten Core-Multishell Nanocarrier drangen nicht in vitale Hautschichten von gesunder oder entzündeter Haut des Imiquimod induzierten Psoriasis ähnlichen Mausmodels in BALB/c Mäusen ein, sondern akkumulierten im Stratum corneum. Vorangegangene in vitro Studien hatten bereits ein Eindringen von Core-Multishell Nanocarriern in humane Haut ex vivo nach Tape stripping gezeigt. Diese Ansammlung der Nanocarrier im Stratum corneum könnte möglicherweise als Depot für eine retardierte und damit verlängerte Wirkstofffreisetzung genutzt werden. Gleichzeitig konnten in den Penetrationsexperimenten keine lokalen oder systemischen negativen Effekte der Core-Multishell Nanocarrier beobachtet werden. Das steht in Einklang mit der vorhandenen Literatur, welche sogar nach wiederholter subkutaner Injektion dieser Nanocarrier keine negativen Effekte beschreibt. Die topische Applikation von mit Nilrot beladenen Core-Multishell Nanocarriern zeigte einen vermehrten Ladungstransport in die vitale Epidermis im Vergleich zu Nilrot in einer Crème. Weitere Aufklärung des Mechanismus, mit dem die Core-Multishell Nanocarrier das Eindringen von Nilrot verstärken, und ein Wirksamkeitsnachweis für echte Wirkstoffe benötigen weiterführende Untersuchungen. Allerdings bleiben die Core-Multishell Nanocarrier vielversprechende Kandidaten für die Weiterentwicklung der Nanocarrier zur Therapie von Hauterkrankungen einschließlich des Nachweises eines Therapievorteils wirkstoffbeladener Core-Multishell Nanocarrier gegenüber kommerziellen topischen Formulierungen. Es konnten keine Tacrolimus spezifischen antiinflammatorischen Effekte im Psoriasis ähnlichen, Imiquimod induzierten Dermatitis Modell in BALB/c Mäusen gezeigt werden, trotz Eindringen des Wirkstoffes in die Dermis. Weitergehende Untersuchungen sind notwendig, um den Grund für die fehlende Effizienz, dessen Widerspruch mit vorhandener Literatur und eine mögliche Abhängigkeit der Tacrolimus Wirksamkeit in diesem Modell von Mausstämmen zu erforschen. Unabhängig davon ist das Imiquimod induzierte Psoriasis Modell mit den hier verwendeten Protokollen in BALB/c Mäusen nicht zur Testung eines möglichen Therapievorteils von Core-Multishell Nanocarriern zum Tacrolimus Transport gegen Tacrolimussalbe geeignet. Des Weiteren wird die Wichtigkeit betont, ein passendes Modell zur Forschungsfragestellung auszuwählen und mehrere objektive Parameter auszuwerten, um Überinterpretation von kleinen Schwankungen gegen alle notwendigen Kontrollgruppen inklusive Vehikelkontrollen zusätzlich zu unbehandelten Kontrollen zu vermeiden

In vitro and in vivo effects of Pelargonium sidoides DC. root extract EPs® 7630 and selected constituents against SARS-CoV-2 B.1, Delta AY.4/AY.117 and Omicron BA.2

The occurrence of immune-evasive SARS-CoV-2 strains emphasizes the importance to search for broad-acting antiviral compounds. Our previous in vitro study showed that Pelargonium sidoides DC. root extract EPs® 7630 has combined antiviral and immunomodulatory properties in SARS-CoV-2-infected human lung cells. Here we assessed in vivo effects of EPs® 7630 in SARS-CoV-2-infected hamsters, and investigated properties of EPs® 7630 and its functionally relevant constituents in context of phenotypically distinct SARS-CoV-2 variants. We show that EPs® 7630 reduced viral load early in the course of infection and displayed significant immunomodulatory properties positively modulating disease progression in hamsters. In addition, we find that EPs® 7630 differentially inhibits SARS-CoV-2 variants in nasal and bronchial human airway epithelial cells. Antiviral effects were more pronounced against Omicron BA.2 compared to B.1 and Delta, the latter two preferring TMPRSS2-mediated fusion with the plasma membrane for cell entry instead of receptor-mediated low pH-dependent endocytosis. By using SARS-CoV-2 Spike VSV-based pseudo particles (VSVpp), we confirm higher EPs® 7630 activity against Omicron Spike-VSVpp, which seems independent of the serine protease TMPRSS2, suggesting that EPs® 7630 targets endosomal entry. We identify at least two molecular constituents of EPs® 7630, i.e., (−)-epigallocatechin and taxifolin with antiviral effects on SARS-CoV-2 replication and cell entry. In summary, our study shows that EPs® 7630 ameliorates disease outcome in SARS-CoV-2-infected hamsters and has enhanced activity against Omicron, apparently by limiting late endosomal SARS-CoV-2 entry

Dendritic Core-Multishell Nanocarriers in Murine Models of Healthy and Atopic Skin

Dendritic hPG-amid-C18-mPEG core-multishell nanocarriers (CMS) represent a novel class of unimolecular micelles that hold great potential as drug transporters, e.g., to facilitate topical therapy in skin diseases. Atopic dermatitis is among the most common inflammatory skin disorders with complex barrier alterations which may affect the efficacy of topical treatment. Here, we tested the penetration behavior and identified target structures of unloaded CMS after topical administration in healthy mice and in mice with oxazolone-induced atopic dermatitis. We further examined whole body distribution and possible systemic side effects after simulating high dosage dermal penetration by subcutaneous injection. Following topical administration, CMS accumulated in the stratum corneum without penetration into deeper viable epidermal layers. The same was observed in atopic dermatitis mice, indicating that barrier alterations in atopic dermatitis had no influence on the penetration of CMS. Following subcutaneous injection, CMS were deposited in the regional lymph nodes as well as in liver, spleen, lung, and kidney. However, in vitro toxicity tests, clinical data, and morphometry- assisted histopathological analyses yielded no evidence of any toxic or otherwise adverse local or systemic effects of CMS, nor did they affect the severity or course of atopic dermatitis. Taken together, CMS accumulate in the stratum corneum in both healthy and inflammatory skin and appear to be highly biocompatible in the mouse even under conditions of atopic dermatitis and thus could potentially serve to create a depot for anti-inflammatory drugs in the skin

Grip strength values and cut-off points based on over 200,000 adults of the German National Cohort - a comparison to the EWGSOP2 cut-off points

BACKGROUND: The European Working Group on Sarcopenia in Older People (EWGSOP) updated in 2018 the cut-off points for low grip strength to assess sarcopenia based on pooled data from 12 British studies. OBJECTIVE: Comparison of the EWGSOP2 cut-off points for low grip strength to those derived from a large German sample. METHODS: We assessed the grip strength distribution across age and derived low grip strength cut-off points for men and women (peak mean -2.5 × SD) based on 200,389 German National Cohort (NAKO) participants aged 19–75 years. In 1,012 Cooperative Health Research in the Region of Augsburg (KORA)-Age participants aged 65–93 years, we calculated the age-standardised prevalence of low grip strength and time-dependent sensitivity and specificity for all-cause mortality. RESULTS: Grip strength increased in the third and fourth decade of life and declined afterwards. Calculated cut-off points for low grip strength were 29 kg for men and 18 kg for women. In KORA-Age, the age-standardised prevalence of low grip strength was 1.5× higher for NAKO-derived (17.7%) compared to EWGSOP2 (11.7%) cut-off points. NAKO-derived cut-off points yielded a higher sensitivity and lower specificity for all-cause mortality. CONCLUSIONS: Cut-off points for low grip strength from German population-based data were 2 kg higher than the EWGSOP2 cut-off points. Higher cut-off points increase the sensitivity, thereby suggesting an intervention for more patients at risk, while other individuals might receive additional diagnostics/treatment without the urgent need. Research on the effectiveness of intervention in patients with low grip strength defined by different cut-off points is needed

Erratum: Global, regional, and national comparative risk assessment of 84 behavioural, environmental and occupational, and metabolic risks or clusters of risks for 195 countries and territories, 1990–2017: a systematic analysis for the Global Burden of Disease Study 2017

Interpretation: By quantifying levels and trends in exposures to risk factors and the resulting disease burden, this assessment offers insight into where past policy and programme efforts might have been successful and highlights current priorities for public health action. Decreases in behavioural, environmental, and occupational risks have largely offset the effects of population growth and ageing, in relation to trends in absolute burden. Conversely, the combination of increasing metabolic risks and population ageing will probably continue to drive the increasing trends in non-communicable diseases at the global level, which presents both a public health challenge and opportunity. We see considerable spatiotemporal heterogeneity in levels of risk exposure and risk-attributable burden. Although levels of development underlie some of this heterogeneity, O/E ratios show risks for which countries are overperforming or underperforming relative to their level of development. As such, these ratios provide a benchmarking tool to help to focus local decision making. Our findings reinforce the importance of both risk exposure monitoring and epidemiological research to assess causal connections between risks and health outcomes, and they highlight the usefulness of the GBD study in synthesising data to draw comprehensive and robust conclusions that help to inform good policy and strategic health planning

The global burden of cancer attributable to risk factors, 2010-19 : a systematic analysis for the Global Burden of Disease Study 2019

Background Understanding the magnitude of cancer burden attributable to potentially modifiable risk factors is crucial for development of effective prevention and mitigation strategies. We analysed results from the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2019 to inform cancer control planning efforts globally. Methods The GBD 2019 comparative risk assessment framework was used to estimate cancer burden attributable to behavioural, environmental and occupational, and metabolic risk factors. A total of 82 risk-outcome pairs were included on the basis of the World Cancer Research Fund criteria. Estimated cancer deaths and disability-adjusted life-years (DALYs) in 2019 and change in these measures between 2010 and 2019 are presented. Findings Globally, in 2019, the risk factors included in this analysis accounted for 4.45 million (95% uncertainty interval 4.01-4.94) deaths and 105 million (95.0-116) DALYs for both sexes combined, representing 44.4% (41.3-48.4) of all cancer deaths and 42.0% (39.1-45.6) of all DALYs. There were 2.88 million (2.60-3.18) risk-attributable cancer deaths in males (50.6% [47.8-54.1] of all male cancer deaths) and 1.58 million (1.36-1.84) risk-attributable cancer deaths in females (36.3% [32.5-41.3] of all female cancer deaths). The leading risk factors at the most detailed level globally for risk-attributable cancer deaths and DALYs in 2019 for both sexes combined were smoking, followed by alcohol use and high BMI. Risk-attributable cancer burden varied by world region and Socio-demographic Index (SDI), with smoking, unsafe sex, and alcohol use being the three leading risk factors for risk-attributable cancer DALYs in low SDI locations in 2019, whereas DALYs in high SDI locations mirrored the top three global risk factor rankings. From 2010 to 2019, global risk-attributable cancer deaths increased by 20.4% (12.6-28.4) and DALYs by 16.8% (8.8-25.0), with the greatest percentage increase in metabolic risks (34.7% [27.9-42.8] and 33.3% [25.8-42.0]). Interpretation The leading risk factors contributing to global cancer burden in 2019 were behavioural, whereas metabolic risk factors saw the largest increases between 2010 and 2019. Reducing exposure to these modifiable risk factors would decrease cancer mortality and DALY rates worldwide, and policies should be tailored appropriately to local cancer risk factor burden. Copyright (C) 2022 The Author(s). Published by Elsevier Ltd. This is an Open Access article under the CC BY 4.0 license.Peer reviewe

Global, regional, and national comparative risk assessment of 84 behavioural, environmental and occupational, and metabolic risks or clusters of risks for 195 countries and territories, 1990-2017: a systematic analysis for the Global Burden of Disease Study 2017

Background The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2017 comparative risk assessment (CRA) is a comprehensive approach to risk factor quantification that offers a useful tool for synthesising evidence on risks and risk–outcome associations. With each annual GBD study, we update the GBD CRA to incorporate improved methods, new risks and risk–outcome pairs, and new data on risk exposure levels and risk–outcome associations. Methods We used the CRA framework developed for previous iterations of GBD to estimate levels and trends in exposure, attributable deaths, and attributable disability-adjusted life-years (DALYs), by age group, sex, year, and location for 84 behavioural, environmental and occupational, and metabolic risks or groups of risks from 1990 to 2017. This study included 476 risk–outcome pairs that met the GBD study criteria for convincing or probable evidence of causation. We extracted relative risk and exposure estimates from 46 749 randomised controlled trials, cohort studies, household surveys, census data, satellite data, and other sources. We used statistical models to pool data, adjust for bias, and incorporate covariates. Using the counterfactual scenario of theoretical minimum risk exposure level (TMREL), we estimated the portion of deaths and DALYs that could be attributed to a given risk. We explored the relationship between development and risk exposure by modelling the relationship between the Socio-demographic Index (SDI) and risk-weighted exposure prevalence and estimated expected levels of exposure and risk-attributable burden by SDI. Finally, we explored temporal changes in risk-attributable DALYs by decomposing those changes into six main component drivers of change as follows: (1) population growth; (2) changes in population age structures; (3) changes in exposure to environmental and occupational risks; (4) changes in exposure to behavioural risks; (5) changes in exposure to metabolic risks; and (6) changes due to all other factors, approximated as the risk-deleted death and DALY rates, where the risk-deleted rate is the rate that would be observed had we reduced the exposure levels to the TMREL for all risk factors included in GBD 2017. Findings In 2017, 34·1 million (95% uncertainty interval [UI] 33·3–35·0) deaths and 1·21 billion (1·14–1·28) DALYs were attributable to GBD risk factors. Globally, 61·0% (59·6–62·4) of deaths and 48·3% (46·3–50·2) of DALYs were attributed to the GBD 2017 risk factors. When ranked by risk-attributable DALYs, high systolic blood pressure (SBP) was the leading risk factor, accounting for 10·4 million (9·39–11·5) deaths and 218 million (198–237) DALYs, followed by smoking (7·10 million [6·83–7·37] deaths and 182 million [173–193] DALYs), high fasting plasma glucose (6·53 million [5·23–8·23] deaths and 171 million [144–201] DALYs), high body-mass index (BMI; 4·72 million [2·99–6·70] deaths and 148 million [98·6–202] DALYs), and short gestation for birthweight (1·43 million [1·36–1·51] deaths and 139 million [131–147] DALYs). In total, risk-attributable DALYs declined by 4·9% (3·3–6·5) between 2007 and 2017. In the absence of demographic changes (ie, population growth and ageing), changes in risk exposure and risk-deleted DALYs would have led to a 23·5% decline in DALYs during that period. Conversely, in the absence of changes in risk exposure and risk-deleted DALYs, demographic changes would have led to an 18·6% increase in DALYs during that period. The ratios of observed risk exposure levels to exposure levels expected based on SDI (O/E ratios) increased globally for unsafe drinking water and household air pollution between 1990 and 2017. This result suggests that development is occurring more rapidly than are changes in the underlying risk structure in a population. Conversely, nearly universal declines in O/E ratios for smoking and alcohol use indicate that, for a given SDI, exposure to these risks is declining. In 2017, the leading Level 4 risk factor for age-standardised DALY rates was high SBP in four super-regions: central Europe, eastern Europe, and central Asia; north Africa and Middle East; south Asia; and southeast Asia, east Asia, and Oceania. The leading risk factor in the high-income super-region was smoking, in Latin America and Caribbean was high BMI, and in sub-Saharan Africa was unsafe sex. O/E ratios for unsafe sex in sub-Saharan Africa were notably high, and those for alcohol use in north Africa and the Middle East were notably low. Interpretation By quantifying levels and trends in exposures to risk factors and the resulting disease burden, this assessment offers insight into where past policy and programme efforts might have been successful and highlights current priorities for public health action. Decreases in behavioural, environmental, and occupational risks have largely offset the effects of population growth and ageing, in relation to trends in absolute burden. Conversely, the combination of increasing metabolic risks and population ageing will probably continue to drive the increasing trends in non-communicable diseases at the global level, which presents both a public health challenge and opportunity. We see considerable spatiotemporal heterogeneity in levels of risk exposure and risk-attributable burden. Although levels of development underlie some of this heterogeneity, O/E ratios show risks for which countries are overperforming or underperforming relative to their level of development. As such, these ratios provide a benchmarking tool to help to focus local decision making. Our findings reinforce the importance of both risk exposure monitoring and epidemiological research to assess causal connections between risks and health outcomes, and they highlight the usefulness of the GBD study in synthesising data to draw comprehensive and robust conclusions that help to inform good policy and strategic health planning

Global, regional, and national comparative risk assessment of 84 behavioural, environmental and occupational, and metabolic risks or clusters of risks for 195 countries and territories, 1990-2017: a systematic analysis for the Global Burden of Disease Study 2017.

BACKGROUND: The Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) 2017 comparative risk assessment (CRA) is a comprehensive approach to risk factor quantification that offers a useful tool for synthesising evidence on risks and risk-outcome associations. With each annual GBD study, we update the GBD CRA to incorporate improved methods, new risks and risk-outcome pairs, and new data on risk exposure levels and risk-outcome associations. METHODS: We used the CRA framework developed for previous iterations of GBD to estimate levels and trends in exposure, attributable deaths, and attributable disability-adjusted life-years (DALYs), by age group, sex, year, and location for 84 behavioural, environmental and occupational, and metabolic risks or groups of risks from 1990 to 2017. This study included 476 risk-outcome pairs that met the GBD study criteria for convincing or probable evidence of causation. We extracted relative risk and exposure estimates from 46 749 randomised controlled trials, cohort studies, household surveys, census data, satellite data, and other sources. We used statistical models to pool data, adjust for bias, and incorporate covariates. Using the counterfactual scenario of theoretical minimum risk exposure level (TMREL), we estimated the portion of deaths and DALYs that could be attributed to a given risk. We explored the relationship between development and risk exposure by modelling the relationship between the Socio-demographic Index (SDI) and risk-weighted exposure prevalence and estimated expected levels of exposure and risk-attributable burden by SDI. Finally, we explored temporal changes in risk-attributable DALYs by decomposing those changes into six main component drivers of change as follows: (1) population growth; (2) changes in population age structures; (3) changes in exposure to environmental and occupational risks; (4) changes in exposure to behavioural risks; (5) changes in exposure to metabolic risks; and (6) changes due to all other factors, approximated as the risk-deleted death and DALY rates, where the risk-deleted rate is the rate that would be observed had we reduced the exposure levels to the TMREL for all risk factors included in GBD 2017