An assessment of the urban water footprint and blue water scarcity: A case study for Van (Turkey) / Uma avaliação da pegada hídrica urbana e da escassez de água azul: um estudo de caso para Van, Turquia

Today, most of the world’s population faces water scarcity, while global warming, urbanization, industrialization and population increases continue to increase the severity of the pressure on water resources. Management ofwater resources plays a key role in the sustainability of agricultural production. The water footprint (WF) is different in comparison to other water statistics because it takes direct and indirect water consumption into account, and helps in the management of water resources. Within this context, the WF of Van province, which is Turkey’s most easterly located arid region, was calculated from 2004 to 2019. The study area covers lake Van, which is Turkey's largest lake, and the Van basin with an area of 23.334km² and a population of 1.136.757 (2019). In the calculations, crop (WFcrop), livestock (WFlivestock), and domestic and industrial water footprints (WFdomestic+industrial) were evaluated separately, and blue and green water footprints (WFblue and WFgreen) were analyzed in detail. According to the results, the average WF of Van province was found to be 8.73 billionm3 year-¹. Throughout the province, 87.6% of the WF is composed of WFcrop, 4.9% is WFlivestock and 7.5% is WFdomestic+industrial. Of the WFcrop, 62.5% depends on WFblue, i.e., freshwater. Most of the WFlivestock consisted of dairy cattle (49%) and sheep (38%). The average WFdomestic+industrial for 2004 to 2019 was 0.64billion m³ year-¹. The average per capita water footprint of Van province was found to be 889.9m³ year-¹ capita-¹. In addition, the province is classified as severe water scarcity (257%). This study is one of the first province-based calculations of WF in Turkey and is the first study to bring a different aspect to published literature by including residual soil moisture from the winter months. As a result of this study, the WFblue of the [...].

Hoje, a maior parte da população mundial enfrenta a escassez de água, enquanto o aquecimento global, a urbanização, a industrialização e o crescimento da população continuam a aumentar a gravidade da pressão sobre os recursos hídricos. A gestão dos recursos hídricos desempenha papel fundamental na sustentabilidade da produção agrícola. A pegada hídrica (WF) é diferente em comparação com outras estatísticas hídricas porque leva em consideração o consumo direto e indireto de água e auxilia na gestão dos recursos hídricos. Nesse contexto, o WF da província de Van, que é a região árida localizada mais a leste da Turquia, foi calculado de 2004 a 2019. A área de estudo cobre o lago Van, que é o maior lago da Turquia, e a bacia de Van, com uma área de 23,334 km² e uma população de 1.136.757 (2019). Nos cálculos, as pegadas hídricas de safra (WFcrop), pecuária (WFlivestock) e doméstica e industrial (WFdomestic+industrial) foram avaliadas separadamente, e as pegadas hídricas azul e verde (WFblue e WFgreen) foram analisadas em detalhes. De acordo com os resultados, o WF médio da província de Van foi encontrado em 8,73 bilhões de m³ ano-¹. Em toda a província, 87,6% do WF são compostos por WFcrop, 4,9% são WFlivestock e 7,5% são WFdomestic+industrial. Do WFcrop, 62,5% dependem do WFblue, ou seja, de água doce. A maior parte do gado WFlivestock era composto por gado leiteiro (49%) e ovelhas (38%). O WFdomestic+industrial médio de 2004 a 2019 foi de 0,64 bilhão de m³ ano-¹. A pegada hídrica per capita média da província de Van foi encontrada em 889,9 m³ ano-¹ capita-¹. Além disso, a região é classificada como grave escassez de água (257%). Este estudo é um dos primeiros cálculos de WF baseados em províncias na Turquia e é o primeiro estudo a trazer um aspecto diferente para a literatura publicada, incluindo a umidade residual do solo dos meses de inverno. Como resultado deste estudo, o WFblue do WFcrop está [...].

An assessment of the urban water footprint and blue water scarcity: A case study for Van (Turkey)

Abstract Today, most of the worlds population faces water scarcity, while global warming, urbanization, industrialization and population increases continue to increase the severity of the pressure on water resources. Management of water resources plays a key role in the sustainability of agricultural production. The water footprint (WF) is different in comparison to other water statistics because it takes direct and indirect water consumption into account, and helps in the management of water resources. Within this context, the WF of Van province, which is Turkeys most easterly located arid region, was calculated from 2004 to 2019. The study area covers lake Van, which is Turkey's largest lake, and the Van basin with an area of 23.334 km2 and a population of 1.136.757 (2019). In the calculations, crop (WFcrop), livestock (WFlivestock), and domestic and industrial water footprints (WFdomestic+industrial) were evaluated separately, and blue and green water footprints (WFblue and WFgreen) were analyzed in detail. According to the results, the average WF of Van province was found to be 8.73 billion m3 year-1. Throughout the province, 87.6% of the WF is composed of WFcrop, 4.9% is WFlivestock and 7.5% is WFdomestic+industrial. Of the WFcrop, 62.5% depends on WFblue, i.e., freshwater. Most of the WFlivestock consisted of dairy cattle (49%) and sheep (38%). The average WFdomestic+industrial for 2004 to 2019 was 0.64 billion m3 year-1. The average per capita water footprint of Van province was found to be 889.9 m3 year-1 capita-1. In addition, the province is classified as severe water scarcity (257%). This study is one of the first province-based calculations of WF in Turkey and is the first study to bring a different aspect to published literature by including residual soil moisture from the winter months. As a result of this study, the WFblue of the WFcrop is above the worldwide average and should be reduced by changing the crop pattern or synchronizing the planting and harvest dates of the crops to a period that benefits from precipitation. In addition, this study is expected to contribute to new studies for calculating the provincial scale WF and will have positive effects on agricultural planning, water allocation and the sustainability of water resources.

Resumo Hoje, a maior parte da população mundial enfrenta a escassez de água, enquanto o aquecimento global, a urbanização, a industrialização e o crescimento da população continuam a aumentar a gravidade da pressão sobre os recursos hídricos. A gestão dos recursos hídricos desempenha papel fundamental na sustentabilidade da produção agrícola. A pegada hídrica (WF) é diferente em comparação com outras estatísticas hídricas porque leva em consideração o consumo direto e indireto de água e auxilia na gestão dos recursos hídricos. Nesse contexto, o WF da província de Van, que é a região árida localizada mais a leste da Turquia, foi calculado de 2004 a 2019. A área de estudo cobre o lago Van, que é o maior lago da Turquia, e a bacia de Van, com uma área de 23,334 km2 e uma população de 1.136.757 (2019). Nos cálculos, as pegadas hídricas de safra (WFcrop), pecuária (WFlivestock) e doméstica e industrial (WFdomestic+industrial) foram avaliadas separadamente, e as pegadas hídricas azul e verde (WFblue e WFgreen) foram analisadas em detalhes. De acordo com os resultados, o WF médio da província de Van foi encontrado em 8,73 bilhões de m3 ano-1. Em toda a província, 87,6% do WF são compostos por WFcrop, 4,9% são WFlivestock e 7,5% são WFdomestic+industrial. Do WFcrop, 62,5% dependem do WFblue, ou seja, de água doce. A maior parte do gado WFlivestock era composto por gado leiteiro (49%) e ovelhas (38%). O WFdomestic+industrial médio de 2004 a 2019 foi de 0,64 bilhão de m3 ano-1. A pegada hídrica per capita média da província de Van foi encontrada em 889,9 m3 ano-1 capita-1. Além disso, a região é classificada como grave escassez de água (257%). Este estudo é um dos primeiros cálculos de WF baseados em províncias na Turquia e é o primeiro estudo a trazer um aspecto diferente para a literatura publicada, incluindo a umidade residual do solo dos meses de inverno. Como resultado deste estudo, o WFblue do WFcrop está acima da média mundial e deve ser reduzido alterando o padrão de cultivo ou sincronizando as datas de plantio e colheita das safras para um período que se beneficie da precipitação. Além disso, espera-se que este estudo contribua para novos estudos para o cálculo da escala provincial WF e terá efeitos positivos no planejamento agrícola, alocação de água e a sustentabilidade dos recursos hídricos.

An assessment of the urban water footprint and blue water scarcity: A case study for Van (Turkey) / Uma avaliação da pegada hídrica urbana e da escassez de água azul: um estudo de caso para Van, Turquia

Today, most of the world's population faces water scarcity, while global warming, urbanization, industrialization and population increases continue to increase the severity of the pressure on water resources. Management of water resources plays a key role in the sustainability of agricultural production. The water footprint (WF) is different in comparison to other water statistics because it takes direct and indirect water consumption into account, and helps in the management of water resources. Within this context, the WF of Van province, which is Turkey's most easterly located arid region, was calculated from 2004 to 2019. The study area covers lake Van, which is Turkey's largest lake, and the Van basin with an area of 23.334km2 and a population of 1.136.757 (2019). In the calculations, crop (WFcrop), livestock (WFlivestock), and domestic and industrial water footprints (WFdomestic+industrial) were evaluated separately, and blue and green water footprints (WFblue and WFgreen) were analyzed in detail. According to the results, the average WF of Van province was found to be 8.73 billionm3 year-1. Throughout the province, 87.6% of the WF is composed of WFcrop, 4.9% is WFlivestock and 7.5% is WFdomestic+industrial. Of the WFcrop, 62.5% depends on WFblue, i.e., freshwater. Most of the WFlivestock consisted of dairy cattle (49%) and sheep (38%). The average WFdomestic+industrial for 2004 to 2019 was 0.64billion m3 year-1. The average per capita water footprint of Van province was found to be 889.9m3 year-1 capita-1. In addition, the province is classified as severe water scarcity (257%). This study is one of the first province-based calculations of WF in Turkey and is the first study to bring a different aspect to published literature by including residual soil moisture from the winter months. As a result of this study, the WFblue of the WFcrop is above the worldwide average and should be reduced by changing the crop pattern or synchronizing the planting and harvest dates of the crops to a period that benefits from precipitation. In addition, this study is expected to contribute to new studies for calculating the provincial scale WF and will have positive effects on agricultural planning, water allocation and the sustainability of water resources.

Hoje, a maior parte da população mundial enfrenta a escassez de água, enquanto o aquecimento global, a urbanização, a industrialização e o crescimento da população continuam a aumentar a gravidade da pressão sobre os recursos hídricos. A gestão dos recursos hídricos desempenha papel fundamental na sustentabilidade da produção agrícola. A pegada hídrica (WF) é diferente em comparação com outras estatísticas hídricas porque leva em consideração o consumo direto e indireto de água e auxilia na gestão dos recursos hídricos. Nesse contexto, o WF da província de Van, que é a região árida localizada mais a leste da Turquia, foi calculado de 2004 a 2019. A área de estudo cobre o lago Van, que é o maior lago da Turquia, e a bacia de Van, com uma área de 23,334 km2 e uma população de 1.136.757 (2019). Nos cálculos, as pegadas hídricas de safra (WFcrop), pecuária (WFlivestock) e doméstica e industrial (WFdomestic+industrial) foram avaliadas separadamente, e as pegadas hídricas azul e verde (WFblue e WFgreen) foram analisadas em detalhes. De acordo com os resultados, o WF médio da província de Van foi encontrado em 8,73 bilhões de m3 ano-1. Em toda a província, 87,6% do WF são compostos por WFcrop, 4,9% são WFlivestock e 7,5% são WFdomestic+industrial. Do WFcrop, 62,5% dependem do WFblue, ou seja, de água doce. A maior parte do gado WFlivestock era composto por gado leiteiro (49%) e ovelhas (38%). O WFdomestic+industrial médio de 2004 a 2019 foi de 0,64 bilhão de m3 ano-1. A pegada hídrica per capita média da província de Van foi encontrada em 889,9 m3 ano-1 capita-1. Além disso, a região é classificada como grave escassez de água (257%). Este estudo é um dos primeiros cálculos de WF baseados em províncias na Turquia e é o primeiro estudo a trazer um aspecto diferente para a literatura publicada, incluindo a umidade residual do solo dos meses de inverno. Como resultado deste estudo, o WFblue do WFcrop está acima da média mundial e deve ser reduzido alterando o padrão de cultivo ou sincronizando as datas de plantio e colheita das safras para um período que se beneficie da precipitação. Além disso, espera-se que este estudo contribua para novos estudos para o cálculo da escala provincial WF e terá efeitos positivos no planejamento agrícola, alocação de água e a sustentabilidade dos recursos hídricos.

An assessment of the urban water footprint and blue water scarcity: A case study for Van (Turkey).

Today, most of the world's population faces water scarcity, while global warming, urbanization, industrialization and population increases continue to increase the severity of the pressure on water resources. Management of water resources plays a key role in the sustainability of agricultural production. The water footprint (WF) is different in comparison to other water statistics because it takes direct and indirect water consumption into account, and helps in the management of water resources. Within this context, the WF of Van province, which is Turkey's most easterly located arid region, was calculated from 2004 to 2019. The study area covers lake Van, which is Turkey's largest lake, and the Van basin with an area of 23.334 km2 and a population of 1.136.757 (2019). In the calculations, crop (WFcrop), livestock (WFlivestock), and domestic and industrial water footprints (WFdomestic+industrial) were evaluated separately, and blue and green water footprints (WFblue and WFgreen) were analyzed in detail. According to the results, the average WF of Van province was found to be 8.73 billion m3 year-1. Throughout the province, 87.6% of the WF is composed of WFcrop, 4.9% is WFlivestock and 7.5% is WFdomestic+industrial. Of the WFcrop, 62.5% depends on WFblue, i.e., freshwater. Most of the WFlivestock consisted of dairy cattle (49%) and sheep (38%). The average WFdomestic+industrial for 2004 to 2019 was 0.64 billion m3 year-1. The average per capita water footprint of Van province was found to be 889.9 m3 year-1 capita-1. In addition, the province is classified as severe water scarcity (257%). This study is one of the first province-based calculations of WF in Turkey and is the first study to bring a different aspect to published literature by including residual soil moisture from the winter months. As a result of this study, the WFblue of the WFcrop is above the worldwide average and should be reduced by changing the crop pattern or synchronizing the planting and harvest dates of the crops to a period that benefits from precipitation. In addition, this study is expected to contribute to new studies for calculating the provincial scale WF and will have positive effects on agricultural planning, water allocation and the sustainability of water resources.

FAST: a randomised phase II study of zolbetuximab (IMAB362) plus EOX versus EOX alone for first-line treatment of advanced CLDN18.2-positive gastric and gastro-oesophageal adenocarcinoma.

BACKGROUND: Claudin 18.2 (CLDN18.2) is contained within normal gastric mucosa epithelial tight junctions; upon malignant transformation, CLDN18.2 epitopes become exposed. Zolbetuximab, a chimeric monoclonal antibody, mediates specific killing of CLDN18.2-positive cells through immune effector mechanisms. PATIENTS AND METHODS: The FAST study enrolled advanced gastric/gastro-oesophageal junction and oesophageal adenocarcinoma patients (aged ≥18 years) with moderate-to-strong CLDN18.2 expression in ≥40% tumour cells. Patients received first-line epirubicin + oxaliplatin + capecitabine (EOX, arm 1, n = 84) every 3 weeks (Q3W), or zolbetuximab + EOX (loading dose, 800 mg/m2 then 600 mg/m2 Q3W) (arm 2, n = 77). Arm 3 (exploratory) was added after enrolment initiation (zolbetuximab + EOX 1000 mg/m2 Q3W, n = 85). The primary endpoint was progression-free survival (PFS) and overall survival (OS) was a secondary endpoint. RESULTS: In the overall population, both PFS [hazard ratio (HR) = 0.44; 95% confidence interval (CI), 0.29-0.67; P < 0.0005] and OS (HR = 0.55; 95% CI, 0.39-0.77; P < 0.0005) were significantly improved with zolbetuximab + EOX (arm 2) compared with EOX alone (arm 1). This significant PFS benefit was retained in patients with moderate-to-strong CLDN18.2 expression in ≥70% of tumour cells (HR = 0.38; 95% CI, 0.23-0.62; P < 0.0005). Significant improvement in PFS was also reported in the overall population of arm 3 versus arm 1 (HR = 0.58; 95% CI, 0.39-0.85; P = 0.0114) but not in high CLDN18.2-expressing patients; no significant improvement in OS was observed in either population. Most adverse events (AEs) related to zolbetuximab + EOX (nausea, vomiting, neutropenia, anaemia) were grade 1-2. Grade ≥3 AEs showed no substantial increases overall (zolbetuximab + EOX versus EOX alone). CONCLUSIONS: In advanced gastric/gastro-oesophageal junction and oesophageal adenocarcinoma patients expressing CLDN18.2, adding zolbetuximab to first-line EOX provided longer PFS and OS versus EOX alone. Zolbetuximab + EOX was generally tolerated and AEs were manageable. Zolbetuximab 800/600 mg/m2 is being evaluated in phase III studies based on clinical benefit observed in the overall population and in patients with moderate-to-strong CLDN18.2 expression in ≥70% of tumour cells.

Investigation of the clonal associations in Acinetobacter Baumannii strains isolated from the respiratory samples of patients in a tertiary research hospital.

OBJECTIVE: The blaOXA resistance genes and ISAba1 were examined in 70 samples from lower respiratory tract of hospitalized patients. MATERIALS AND METHODS: Of the 67 isolates obtained, almost half (46.3%) of them were from endotracheal aspirate, and most were collected from the intensive care units of the reanimation (37.3%) and internal medicine (32.8%) units. RESULTS: Three samples from the internal medicine intensive care unit had positive cultures. Of the multidrug resistant (MDR) samples, 70 isolates (>50%) were moderately sensitive, while fewer (10%) were resistant to tigecycline. In contrast, 100% were sensitive to colistin. All strains were found to be positive for blaOXA-23-like and blaOXA-51-like genes, whereas no blaOXA-40-like and blaOXA-58-like genes were detected. The ISAba1 positivity rate was 90.0%. Pattern 5 was mainly identified among the 22 different patterns. Of note, 50% of Pattern 5 was found in the patients of the internal medicine intensive care unit, and a third was associated with ventilator-associated pneumonia. Importantly, the internal medicine unit's equipment was found to be culture positive. CONCLUSION: Findings obtained from this study suggest that isolates can easily spread through the hospital via isolate cross-contamination caused by health personnel. These contaminating isolates may be able to maintain their presence within the hospital for a long time.

A multicentre, phase IIa study of zolbetuximab as a single agent in patients with recurrent or refractory advanced adenocarcinoma of the stomach or lower oesophagus: the MONO study.

BACKGROUND: Claudin 18.2 (CLDN18.2) is physiologically confined to gastric mucosa tight junctions; however, upon malignant transformation, perturbations in cell polarity lead to CLDN18.2 epitopes being exposed on the cancer cell surface. The first-in-class monoclonal antibody, zolbetuximab (formerly known as IMAB362), binds to CLDN18.2 and can induce immune-mediated lysis of CLDN18.2-positive cells. PATIENTS AND METHODS: Patients with advanced gastric, gastro-oesophageal junction (GEJ) or oesophageal adenocarcinomas with moderate-to-strong CLDN18.2 expression in ≥50% of tumour cells received zolbetuximab intravenously every 2 weeks for five planned infusions. At least three patients were enrolled in two sequential cohorts (cohort 1300 mg/m2; cohort 2600 mg/m2); additional patients were enrolled into a dose-expansion cohort (cohort 3600 mg/m2). The primary end point was the objective response rate [ORR: complete and partial response (PR)]; secondary end points included clinical benefit [ORR+stable disease (SD)], progression-free survival, safety/tolerability, and zolbetuximab pharmacokinetic profile. RESULTS: From September 2010 to September 2012, 54 patients were enrolled (cohort 1, n = 4; cohort 2, n = 6; cohort 3, n = 44). Three patients in cohort 1 and 25 patients in cohorts 2/3 received at least 5 infusions. Antitumour activity data were available for 43 patients, of whom 4 achieved PR (ORR 9%) and 6 (14%) had SD for a clinical benefit rate of 23%. In a subgroup of patients with moderate-to-high CLDN18.2 expression in ≥70% of tumour cells, ORR was 14% (n = 4/29). Treatment-related adverse events occurred in 81.5% (n = 44/54) patients; nausea (61%), vomiting (50%), and fatigue (22%) were the most frequent. CONCLUSIONS: Zolbetuximab monotherapy was well tolerated and exhibited antitumour activity in patients with CLDN18.2-positive advanced gastric or GEJ adenocarcinomas, with response rates similar to those reported for single-agent targeted agents in gastric/GEJ cancer trials. CLINICALTRIALS.GOV NUMBER: NCT01197885.

Investigation of charge ratio variation in mRNA - DEAE-dextran polyplex delivery systems.

mRNA pharmaceuticals represent a new class of therapeutics, with applications, in cancer vaccination, tumour therapy and protein substitution. Formulations are required to deliver messenger RNA (mRNA) to the target sites where induction of genetic transfection following receptor mediated cell uptake & translation is required. In the current study, the cationic polysaccharide diethylaminoethylen (DEAE) - Dextran was selected as a model system carrier for the investigation of polyplex nanoparticle formation together with mRNA as a function of the molar ratio of the components. The structure of the mRNA/Dextran colloids was investigated as a function of the polymer-to-mRNA ratio and correlated with the biological activity determined by cellular transfection with luciferase coding mRNA. Dynamic light scattering (DLS), small angle x-ray scattering (SAXS), and small angle neutron scattering (SANS) with deuterium contrast variation were used to achieve structural insight into the systems. Similarly to previously investigated lipid based systems, colloidally stable particles with confined size were obtained with either excess of positive or negative charge. Highest activity was obtained with positive charge excess. From the scattering experiments information on the internal organization inside the polymer/mRNA systems was derived. Indication for the presence of structural elements in the length scale of ten to 20 nm were found in the excess of dextran, which could be due to either excess or particulate polymer. Information on the molecular organization of the mRNA nanoparticle products may provide a valuable basis for defining critical quality attributes of drug products for pharmaceutical application.

Actively personalized cancer vaccines--the step into clinical application.

Cancer vaccine development enters a new phase of innovation based on the development of modern sequencing technologies and novel RNA-based synthetic drug formats which enable the analysis and therapeutic targeting of every patient's tumor genome. By applying and combining these innovations, we have brought the concept of "actively personalized cancer vaccines" to clinical testing. Synthetic RNA is used as the drug format, allowing affordable, individual "on demand" manufacturing of tumor-optimized vaccines.

Translation of genomics-guided RNA-based personalised cancer vaccines: towards the bedside.

Cancer is a disease caused by DNA mutations. Cancer therapies targeting defined functional mutations have shown clinical benefit. However, as 95% of the mutations in a tumour are unique to that single patient and only a small number of mutations are shared between patients, the addressed medical need is modest. A rapidly determined patient-specific tumour mutation pattern combined with a flexible mutation-targeting drug platform could generate a mutation-targeting individualised therapy, which would benefit each single patient. Next-generation sequencing enables the rapid identification of somatic mutations in individual tumours (the mutanome). Immunoinformatics enables predictions of mutation immunogenicity. Mutation-targeting RNA-based vaccines can be rapidly and affordably synthesised as custom GMP drug products. Integration of these cutting-edge technologies into a clinically applicable process holds the promise of a disruptive innovation benefiting cancer patients. Here, we describe our translation of the individualised RNA-based cancer vaccine concept into clinic trials.

T-cell receptor transfer into human T cells with ecotropic retroviral vectors.

Adoptive T-cell transfer for cancer immunotherapy requires genetic modification of T cells with recombinant T-cell receptors (TCRs). Amphotropic retroviral vectors (RVs) used for TCR transduction for this purpose are considered safe in principle. Despite this, TCR-coding and packaging vectors could theoretically recombine to produce replication competent vectors (RCVs), and transduced T-cell preparations must be proven free of RCV. To eliminate the need for RCV testing, we transduced human T cells with ecotropic RVs so potential RCV would be non-infectious for human cells. We show that transfection of synthetic messenger RNA encoding murine cationic amino-acid transporter 1 (mCAT-1), the receptor for murine retroviruses, enables efficient transient ecotropic transduction of human T cells. mCAT-1-dependent transduction was more efficient than amphotropic transduction performed in parallel, and preferentially targeted naive T cells. Moreover, we demonstrate that ecotropic TCR transduction results in antigen-specific restimulation of primary human T cells. Thus, ecotropic RVs represent a versatile, safe and potent tool to prepare T cells for the adoptive transfer.

Selective uptake of naked vaccine RNA by dendritic cells is driven by macropinocytosis and abrogated upon DC maturation.

Even though it is known for more than one decade that antigen-encoding RNA can deliver antigenic information to induce antigen-specific immunity against cancer, the nature and mechanism of RNA uptake have remained enigmatic. In this study, we investigated the pharmacokinetics of naked RNA administered into the lymph node. We observed that RNA is rapidly and selectively uptaken by lymph node dendritic cells (DCs). Furthermore, in vitro and in vivo studies revealed that the efficient internalization of RNA by human and murine DCs is primarily driven by macropinocytosis. Selective inhibition of macropinocytosis by compounds or as a consequence of DC maturation abrogated RNA internalization and delivery of encoded antigens. Our findings imply that bioavailability of recombinant RNA vaccines in vivo highly depends on the density and the maturation stage of DCs at the administration site and are of importance for the design of RNA-based clinical immunotherapy protocols.

Tumor-associated CpG demethylation augments hypoxia-induced effects by positive autoregulation of HIF-1α.

Hypoxia-inducible factor 1α (HIF-1α) is frequently overexpressed in human cancers and controls the expression of several genes that have been implicated in tumor growth and progression. Activity of HIF-1α in cancer cells is regulated at the transcriptional, translational and posttranslational level by multiple inter- and coacting molecular pathways. In this report, we reveal for the first time that tumor-associated CpG demethylation facilitates positive autoregulation of HIF-1α, resulting in amplification of hypoxia-induced transactivation of HIF-1α target genes. The HIF-1α promoter harbors a hypoxia response element that is normally repressed by methylation of a CpG dinucleotide located in the core element. In colon cancer cell lines and in primary colon cancer specimens, however, we found frequent aberrant demethylation of this element, enabling binding of HIF-1α to its own promoter resulting in autotransactivation of HIF-1α expression. Our results provide novel and highly unexpected insights into the complexity of HIF-1α regulation in cancer cells and implicate that tumor-associated CpG demethylation augments HIF-1α-mediated effects on malignant cell growth.

Phosphorothioate cap analogs increase stability and translational efficiency of RNA vaccines in immature dendritic cells and induce superior immune responses in vivo.

Vaccination with in vitro transcribed RNA coding for tumor antigens is considered a promising approach for cancer immunotherapy and has already entered human clinical testing. One of the basic objectives for development of RNA as a drug is the optimization of immunobioavailability of the encoded antigen in vivo. By analyzing the effect of different synthetic 5' mRNA cap analogs on the kinetics of the encoded protein, we found that m(2)(7,2'-O)Gpp(S)pG (beta-S-ARCA) phosphorothioate caps, in particular the D1 diastereoisomer, profoundly enhance RNA stability and translational efficiency in immature but not mature dendritic cells. Moreover, in vivo delivery of the antigen as beta-S-ARCA(D1)-capped RNA species is superior for protein expression and for efficient priming and expansion of naïve antigen-specific T cells in mice. Our findings establish 5' mRNA cap analogs as yet another module for tuning immunopharmacological properties of recombinant antigen-encoding RNA for vaccination purposes.

Prevalence and Risk Factors of Allergies in Turkey (PARFAIT): results of a multicentre cross-sectional study in adults.

The Prevalence and Risk Factors of Allergies in Turkey (PARFAIT) study was planned to evaluate the prevalence of and risk factors for asthma and allergic diseases in Turkey. The present analysis used data from 25,843 parents of primary school children, obtained from a cross-sectional questionnaire-based study. A total of 25,843 questionnaires from 14 centres were evaluated. In rural areas, the prevalences asthma, wheezing, allergic rhinitis and eczema in males were: 8.5% (95% confidence interval (CI) 7.9-9.1%), 13.5% (95% CI 12.8-14.2%), 17.5% (95% CI 16.7-18.2%) and 10.8% (95% CI 10.2-11.4%), respectively; and in females were: 11.2% (95% CI 10.9-11.8%), 14.7% (95% CI 14.3-15.1%), 21.2% (95% CI 20.4-22.0%) and 13.1% (95% CI 12.4-13.8%), respectively. In urban areas, the corresponding prevalences in males were: 6.2% (95% CI 5.8-6.6%), 10.8% (95% CI 10.3-11.3%), 11.7% (95% CI 11.4-12.0%) and 6.6% (95% CI 6.2-7.0%), respectively; and in females were: 7.5 % (95% CI 7.9-7.1%), 12.0% (95% CI 11.7-12.3%), 17.0% (95% CI 16.4-17.6%) and 7.3% (95% CI 6.9-7.7%), respectively. Having an atopic first-degree relative or any other atopic diseases had significant effects on the prevalence of allergic diseases. Housing conditions, such as living in a shanty-type house, visible moulds at home and use of wood or biomass as heating or cooking material were associated with one or more allergic diseases. Although genetic susceptibility is strongly associated, country- and population-based environmental factors may contribute to increased prevalence rates of allergic diseases.