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Purpose Refugees’ access to psychotherapeutic care is insufficient in Germany. One factor particularly contributing to hindering their access to adequate therapeutic care is a lack of provision of language mediation. This paper aims to explore the institutional system in which the financing of language mediation in the context of the medical treatment of asylum seekers in Germany is located. It examines why the language barrier problem resulting from a lack of financing is not being solved, even though it has been well known for years as a structural problem of day-patient health care to refugees and migrants in Germany. Design/methodology/approach The financing of language mediation is analysed against the background of theories of the so-called “shunting yard”, in which public responsibilities for the assumption of costs are shifted from one level and actor to the other, thus preventing sustainable solutions being achieved. A mix of qualitative methods including the evaluation of official documents, reports and secondary literature, and of 23 expert interviews was used. Findings The financing of language mediation is a perfect example of the “shunting yard” phenomenon, with responsibilities being shifted between federal government, health insurance bodies and the municipalities in Germany. This paper argues that the specific financing structure in the German federal system can be viewed as a reason for the non-solution of the language barrier that hinders refugees’ access to health care. Originality/value The problem of the financing of language mediation in the context of health care has been rarely treated from a social sciences perspective. This paper contributes to addressing this gap.

Sprachmittlung ist für Chancengleichheit beim Zugang zum Gesundheitssystem von großer praktischer Bedeutung, findet jedoch in der politik- und sozialwissenschaftlichen Forschung bislang kaum Beachtung. Die Bedeutung von Sprachmittlung hat sich zuletzt insbesondere beim Zugang von Menschen mit Fluchterfahrung zu psychotherapeutischer Gesundheitsversorgung gezeigt, die – in einem noch viel höheren Maß als die Versorgung rein körperlicher Erkrankungen – von einer gemeinsamen sprachlichen Verständigungsebene abhängt. In diesem Artikel fragen wir danach, inwieweit deutsche Kommunen Sprachmittlung für Asylbewerber*innen, die eine Psychotherapie aufnehmen möchten, finanzieren. Wir stellen dabei anknüpfend an theoretische Erkenntnisse aus der Implementations- und der Kommunalforschung Faktoren dar, die zum Verständnis der kommunalen (Nicht-)Finanzierung von Sprachmittlung beitragen. Auf Grundlage einer deutschlandweiten Befragung kommen wir zu dem Ergebnis, dass bei der Sprachmittlungsfinanzierung eine große kommunale Varianz vorherrscht, wodurch der Zugang zu dieser Leistung – und damit auch zur psychotherapeutischen Versorgung – zu einer wohnortabhängigen Glückssache wird. Ursächlich hierfür sind insbesondere die klassischen, von Lipsky identifizierten Rahmenbedingungen. So hängt die Entscheidung über Sprachmittlungsfinanzierungsanträge häufig vom Wissen sowie den individuellen Einstellungen der Sachbearbeitenden auf der kommunalen Entscheidungsebene ab.

Globally, construction of dams has led to challenges for fishery managers and decision makers. Hundreds of thousands of dams, many of which no longer serve their intended purpose, are in need of repair. Resources to make those repairs are limited, and dam removal often seems like the most logical solution from an economic perspective. However, dams on the Laurentian Great Lakes tributaries often serve more than one purpose, with nearly 500 considered important to the continued success of controlling sea lamprey (Petromyzon marinus), blocking other invasive species, isolating native from non-native salmonids, stopping disease transfer, and holding back contaminated sediments. Removing dams can have unintended consequences at the local, regional, or basin-wide scale. Here, we explain the importance of considering potential fishery management trade-offs of barrier removals at those scales. We also suggest an organizational framework that, when supported by modeling, could improve communication and cooperation among partners, streamline the decision process, and provide a consensus-driven perspective about the highest priority projects to address. Consistent communication among and between management agencies, indigenous peoples, and local governments, along with an objective and proactive approach to barrier removal decisions could allow for greater success in habitat restoration and funding procurement while reducing the risk of barrier failures and the unintended spread of injurious invasive species, environmental contaminants, and fish disease.

Відносини між Україною та Європейським Союзом (ЄС) привертають все більшу увагу міжнародної спільноти в контексті війни Росії проти України. Використовуючи дані, які були зібрані в рамках Панельного дослідження переселення українців (Resettlement of Ukrainians Panel Study, ReUP), в цьому робочому документі під назвою DeZIM.insights ми аналізуємо чинники трьох аспектів підтримки курсу на ЄС: позитивне ставлення, зв'язок і підтримка вступу до ЄС. Ми співвідносимо підтримку з різними поняттями, наприклад, з громадянською ідентичністю, а також із соціально-демографічними змінними, як-от вік, освіта та стать. Ми спостерігаємо серед українців високий рівень підтримки вступу до ЄС та позитивне загальне ставлення до ЄС. Наш багатовимірний аналіз показує, що саме громадянська ідентичність і регіональне походження виступають чинниками орієнтації на ЄС. У розрізі соціально-демографічних змінних ми спостерігаємо вищі рівні підтримки серед найстаршої вікової групи (респонденти віком від 51 року) у всіх трьох аспектах, а також вищий рівень усвідомлення зв'язку із ЄС серед респондентів з вищою освітою. Крім того, виказуючи меншу підтримку в контексті ставлення та зв'язку, жінки більш рішуче виступають за вступ до ЄС, ніж чоловіки.

Отношения между Украиной и Европейским союзом (ЕС) привлекают все большее внимание международного сообщества в контексте войны России против Украины. Используя данные, собранные в рамках Панельного исследования переселения украинцев (ReUP), мы анализируем в этом рабочем документе DeZIM.insights факторы трех аспектов поддержки ЕС: позитивное отношение, привязанность и поддержка вступления в ЕС. Мы связываем поддержку с различными понятиями, такими как общинная идентичность, и с социально-демографическими переменными, такими как возраст, образование и пол. Мы находим, что поддержка вступления в ЕС среди украинцев высока и что общее отношение к ЕС положительное. Наш многофакторный анализ показывает, что общинная идентичность и, в частности, региональное происхождение, по-видимому, являются движущими силами ориентации на ЕС. Что касается социально-демографических переменных, мы обнаруживаем более высокий уровень поддержки среди самой старшей возрастной группы (респонденты в возрасте 51 года и старше) на всех трех уровнях и более высокое чувство привязанности к ЕС среди респондентов с высшим образованием. Более того, демонстрируя меньшую поддержку через отношение и привязанность, женщины более решительно выступают за вступление в ЕС, чем мужчины.

Werden die Menschen in Deutschland aufgrund der Inflation und der massiv steigenden Energiepreise im Herbst und Winter 2022 massenhaft auf die Straße gehen? Drohen, wie vielfach befürchtet, ein „heißer Herbst“ oder „Wutwinter“ und eine „Querfront“ aus linken und rechten Akteur*innen? Auf Basis einer repräsentativen Bevölkerungsbefragung des DeZIM.panels gehen wir den Fragen nach, wie groß das Protestpotenzial aktuell ist und wie dieses Potenzial mit soziodemographischen Merkmalen und politischen Faktoren zusammenhängt. Unsere Analysen zeigen, dass sich rund jede*r vierte Befragte vorstellen kann, aufgrund der hohen Energiepreise zu protestieren. Etwas über die Hälfte will deswegen nicht auf die Straße gehen, knapp jede*r Fünfte ist noch unentschlossen. Bei Personen, die die AfD wählen würden, ist das Protestpotenzial fast doppelt so hoch wie in der Gesamtbevölkerung. Darüber hinaus dokumentieren die Daten, dass diejenigen Befragten, die in der Vergangenheit gegen die Corona-Maßnahmen protestiert haben, dreimal so häufig zum Protest in der Energiekrise bereit sind als der Rest der Bevölkerung

Introduction Children with Down syndrome (DS) are at increased risk of obstructive sleep disordered breathing (SDB), which is associated with sleep disruption affecting daytime functioning. We examined the effects of treatment of SDB on sleep and daytime functioning in children with DS. Methods Children with DS and SDB (n=34) completed a baseline and follow-up study which included 7 days of actigraphy in conjunction with a parental sleep diary. Parents also completed a number of questionnaires assessing sleep, behaviour, daytime functioning and quality of life (QOL). All children had overnight polysomnography (PSG) at baseline and 24 had PSG at follow-up. Results 15 children (44%) were treated. At baseline the treated group had more severe SDB at baseline compared to the untreated group: obstructive apnoea hypopnoea index (OAHI) 29.3 ± 38.2 events/h vs 3.3 ± 5.2 events/h (p< 0.01). Actigraphy showed no significant differences in total sleep time or sleep efficiency from baseline to follow up in either the treated or untreated group. Wake after sleep onset increased at follow-up in the untreated group (p< 0.01). The sleep disturbance (p< 0.01) and total problems (p< 0.05) scales on the OSA-18 and the SDB subscale on the Pediatric Sleep Problem Survey Instrument (p< 0.01) improved in the treated children. No changes were seen in any of the measures in the untreated children. Conclusion Treatment of SDB improves QOL, despite treatment having no demonstrable impacts on actigraphic sleep measures. In contrast despite having less severe SDB children who were untreated had no improvements in QOL and increased sleep disruption. Support (if any) Jerome Lejeune Foundation

The relationship between Ukraine and the European Union (EU) has received increasing international attention in the context of Russia’s war against Ukraine. Using data collected as part of the Resettlement of Ukrainians Panel Study (ReUP), we analyse in this DeZIM.insights working paper the drivers of three dimensions of support towards the EU: positive attitude, attachment, and support for EU accession. We relate support with different concepts such as communal identity and with sociodemographic variables such as age, education, and gender. We find that support among Ukrainians for EU accession is high and that overall attitudes towards the EU are positive. Our multivariate analyses reveal that communal identity and regional origins in particular appear to be drivers of an orientation towards the EU. Regarding sociodemographic variables, we find higher levels of support among the oldest age group (respondents aged 51 years and older) on all three levels, and a higher feeling of attachment with the EU among respondents with a university degree.

HDV infection causes severe liver disease, the global health burden of which may be underestimated due to limited epidemiological data. HDV depends on HBV for infection, but recent studies indicated that dissemination can also be supported by other helper viruses such as HCV. We used a rapid point-of-care test and an ELISA to retrospectively test for antibodies against the Hepatitis Delta antigen (anti-HDV-Ab) in 4103 HBsAg-positive and 1661 HBsAg-negative, anti-HCV-positive sera from China and Germany. We found that the HDV seroprevalence in HBsAg-positive patients in China is limited to geographic hotspots (Inner Mongolia: 35/251, 13.9%, Xinjiang: 7/180, 3.9%) and high-risk intravenous drug users (HBV mono-infected: 23/247, 9.3%, HBV-HCV co-infected: 34/107, 31.8%), while none of the 2634 HBsAg carriers from other metropolitan regions were anti-HDV-Ab-positive. In Germany, we recorded an HDV seroprevalence of 5.3% in a university hospital environment. In a cohort of HBsAg-negative, anti-HCV-positive patients that were not exposed to HBV before (anti-HBc-negative), HDV was not associated with HCV mono-infection (Chinese high-risk cohort: 0/365, 0.0%, German mixed cohort: 0/263, 0.0%). However, 21/1033 (2.0%) high-risk HCV patients in China with markers of a previously cleared HBV infection (anti-HBc-positive) were positive for anti-HDV-Ab, with two of them being positive for both HDV and HCV RNA but negative for HBV DNA. The absence of anti-HDV-Ab in HCV mono-infected patients shows that HCV cannot promote HDV transmission in humans.

Web-based decision support tools (DSTs) can be useful to facilitate decision-making processes for managing complex natural resource systems. However, the alignment of DSTs with the objectives in governmental policies or management plans and the influence of limited local data on the outputs of these tools may reduce the use of DSTs by decision makers. In this study, we examined the outcomes of web-based DSTs when different types of local data were incorporated and demonstrated a way to incorporate outputs from multiple DSTs or local inventories to benefit barrier removal decisions. Restoring habitat connectivity in rivers in northwest lower Michigan, USA, was used as a case study due to the abundance of local inventory data and web-based DSTs. We found that, when compared to prioritizations made using local data, some DSTs could produce similar outcomes (in barriers selected, cost, and the benefit for migratory fish) with limited data, but the trade-offs among users' objectives might influence the cost and effectiveness of DSTs' outputs. Improving the ability of DSTs to incorporate objectives consistent with policy and stakeholders' values (e.g., restore certain species or sedimentation control) across management scales can help close the gap between tool recommendations and management decisions while making the barrier removal prioritization process transparent and efficient.

Background & Aims Chronic HDV infections cause the most severe form of viral hepatitis. HDV requires HBV envelope proteins for hepatocyte entry, particle assembly and release. Eight HDV and 8 HBV genotypes have been identified. However, there are limited data on the replication competence of different genotypes and the effect that different HBV envelopes have on virion assembly and infectivity. Methods We subcloned complementary DNAs (cDNAs) of all HDV and HBV genotypes and systematically studied HDV replication, assembly and infectivity using northern blot, western blot, reverse-transcription quantitative PCR, and in-cell ELISA. Results The 8 HDV cDNA clones initiated HDV replication with noticeable differences regarding replication efficacy. The 8 HBV-HBsAg-encoding constructs all supported secretion of subviral particles, however variations in envelope protein stoichiometry and secretion efficacy were observed. Co-transfection of all HDV/HBV combinations supported particle assembly, however, the respective pseudo-typed HDVs differed with respect to assembly kinetics. The most productive combinations did not correlate with the natural geographic distribution, arguing against an evolutionary adaptation of HDV ribonucleoprotein complexes to HBV envelopes. All HDVs elicited robust and comparable innate immune responses. HBV envelope-dependent differences in the activity of the EMA-approved entry inhibitor bulevirtide were observed, however efficient inhibition could be achieved at therapeutically applied doses. Lonafarnib also showed pan-genotypic activity. Conclusions HDVs from different genotypes replicate with variable efficacies. Variations in HDV genomes and HBV envelope proteins are both major determinants of HDV egress and entry efficacy, and consequently assembly inhibition by lonafarnib or entry inhibition by bulevirtide. These differences possibly influence HDV pathogenicity, immune responses and the efficacy of novel drug regimens. Lay summary HDV requires the envelope protein of HBV for assembly and to infect human cells. We investigated the ability of different HDV genotypes to infect cells and replicate. We also assessed the effect that envelope proteins from different HBV genotypes had on HDV infectivity and replication. Herein, we confirmed that genotypic differences in HDV and HBV envelope proteins are major determinants of HDV assembly, de novo cell entry and consequently the efficacy of novel antivirals.

Background/aim ALK inhibitors like Crizotinib, Ceritinib and Alectinib are targeted therapies used in patients with anaplastic lymphoma kinase (ALK)-positive, advanced non-small cell lung cancer (NSCLC). Since in this tumor entity radiotherapy is employed sequentially or concomitantly, potential synergistic effects were investigated, which may support the hypothesis of induced radiosensitization by using ALK inhibitors. Materials and methods Two cell lines expressing wild-type (WT) or echinoderm microtubule-associated protein-like 4 (EML4)-ALK were treated with ALK inhibitors, followed by irradiation. Cell survival, cell death, cell cycle and phosphorylation of H2A histone family, member X (H2AX) were examined. Results Combined treatment with ALK-inhibitors plus 10 Gy-irradiation led to effects similar to those of sole radiotherapy, but was more effective than sole drug treatment. Conclusion There is no clear evidence of sensitization to radiation by treating EML4-ALK mutated cells with ALK inhibitors.

Cellular phenotypes are established and controlled by complex and precisely orchestrated molecular networks. In cancer, mutations and dysregulations of multiple molecular factors perturb the regulation of these networks and lead to malignant transformation. High-throughput technologies are a valuable source of information to establish the complex molecular relationships behind the emergence of malignancy, but full exploitation of this massive amount of data requires bioinformatics tools that rely on network-based analyses. In this report we present the Virtual Melanoma Cell, an online tool developed to facilitate the mining and interpretation of high-throughput data on melanoma by biomedical researches. The platform is based on a comprehensive, manually generated and expert-validated regulatory map composed of signaling pathways important in malignant melanoma. The Virtual Melanoma Cell is a tool designed to accept, visualize and analyze user-generated datasets. It is available at: https://www.vcells.net/melanoma . To illustrate the utilization of the web platform and the regulatory map, we have analyzed a large publicly available dataset accounting for anti-PD1 immunotherapy treatment of malignant melanoma patients.

Controlling invasive species is critical for conservation but can have unintended consequences for native species and divert resources away from other efforts. This dilemma occurs on a grand scale in the North American Great Lakes, where dams and culverts block tributary access to habitat of desirable fish species and are a lynchpin of long-standing efforts to limit ecological damage inflicted by the invasive, parasitic sea lamprey (Petromyzon marinus). Habitat restoration and sea-lamprey control create conflicting goals for managing aging infrastructure. We used optimization to minimize opportunity costs of habitat gains for 37 desirable migratory fishes that arose from restricting sea lamprey access (0-25% increase) when selecting barriers for removal under a limited budget (US$1-105 million). Imposing limits on sea lamprey habitat reduced gains in tributary access for desirable species by 15-50% relative to an unconstrained scenario. Additional investment to offset the effect of limiting sea-lamprey access resulted in high opportunity costs for 30 of 37 species (e.g., an additional US$20-80 million for lake sturgeon [Acipenser fulvescens]) and often required ≥5% increase in sea-lamprey access to identify barrier-removal solutions adhering to the budget and limiting access. Narrowly distributed species exhibited the highest opportunity costs but benefited more at less cost when small increases in sea-lamprey access were allowed. Our results illustrate the value of optimization in limiting opportunity costs when balancing invasion control against restoration benefits for diverse desirable species. Such trade-off analyses are essential to the restoration of connectivity within fragmented rivers without unleashing invaders.

Rapid evolution of pest, pathogen, and wildlife populations can have undesirable effects, for example, when insects evolve resistance to pesticides or fishes evolve smaller body size in response to harvest. A destructive invasive species in the Laurentian Great Lakes, the sea lamprey (Petromyzon marinus) has been controlled with the pesticide 3-trifluoromethyl-4-nitrophenol (TFM) since the 1950s. We evaluated the likelihood of sea lamprey evolving resistance to TFM by (i) reviewing sea lamprey life history and control; (ii) identifying physiological and behavioural resistance strategies; (iii) estimating the strength of selection from TFM; (iv) assessing the timeline for evolution; and (v) analyzing historical toxicity data for evidence of resistance. The number of sea lamprey generations exposed to TFM was within the range observed for fish populations where rapid evolution has occurred. Mortality from TFM was estimated as 82%–90%, suggesting significant selective pressure. However, 57 years of toxicity data revealed no increase in lethal concentrations of TFM. Vigilance and the development of alternative controls are required to prevent this aquatic invasive species from evolving strategies to evade control.

Background Equine gastrointestinal nematodes (GINs) have been the subject of intermittent studies in Australia over the past few decades. However, comprehensive information on the epidemiology of equine GINs, the efficacy of available anthelmintic drugs and the prevalence of anthelmintic resistance (AR) in Australasia is lacking. Herein, we have systematically reviewed existing knowledge on the horse GINs recorded in Australia, and main aspects of their pathogeneses, epidemiology, diagnoses, treatment and control. Methods Six electronic databases were searched for publications on GINs of Australian horses that met our inclusion criteria for the systematic review. Subsets of publications were subjected to review epidemiology, diagnoses, pathogeneses, treatment and control of GINs of horses from Australia. Results A total of 51 articles published between 1950 to 2018 were included. The main GINs reported in Australian horses were cyathostomins (at least 28 species), Draschia megastoma, Habronema muscae, H. majus, Oxyuris equi, Parascaris equorum, Strongyloides westeri and Trichostrongylus axei across different climatic regions of Queensland, New South Wales, Victoria, and Western Australia. Nematodes are diagnosed based on the traditional McMaster egg counting technique, though molecular markers to characterise common GINs of equines were characterised in 1990s. The use of anthelmintic drugs remains the most widely-used strategy for controlling equine GIN parasites in Australia; however, the threshold of faecal egg count that should trigger treatment in horses, remains controversial. Furthermore, anthelmintic resistance within GIN population of horses is becoming a common problem in Australia. Conclusions Although GINs infecting Australian horses have been the subject of occasional studies over the past few decades, the effective control of GIN infections is hampered by a generalised lack of knowledge in various disciplines of equine parasitology. Therefore, coordinated and focused research is required to fill our knowledge gaps in these areas to maximise equine health and minimise economic losses associated with the parasitic infections in Australia. Electronic supplementary material The online version of this article (10.1186/s13071-019-3445-4) contains supplementary material, which is available to authorized users.

Background/aim Recent evidence suggests that melanoma patients treated with BRAF inhibitors experience radiosensitization with an increased frequency of side-effects. This could also imply increased effectiveness when treating melanoma. Materials and methods To test whether the BRAF inhibitors dabrafenib and vemurafenib together with ionizing radiation more effectively inhibit melanoma cells, primary human melanoma tumor cell lines expressing wild-type (WT) or mutant V600E BRAF were analyzed by cell survival, cell death, and cell-cycle testing. Results All melanoma cell lines examined were radioresistant in these assays. BRAF inhibitor treatment alone suppressed cell survival more effectively than radiation in all the mutant V600E BRAF cell lines, and vemurafenib, but not dabrafenib, also inhibited cell survival in the WT BRAF cell lines at clinically relevant concentrations. However, when cells were treated with BRAF inhibitor followed by radiation, there was no increased effect on the suppression of cell survival. Vemurafenib induced more necrosis than radiation in most melanoma cell lines, irrespective of BRAF status, but this effect was not additive with the combination treatment. BRAF inhibitors and radiation had variable, but independent effects on the induction of cell-cycle arrest. Conclusion These results suggest that BRAF inhibitors and ionizing radiation do not act synergistically to inhibit the growth of primary human melanoma cells.

Administering immunoregulatory cells as medicinal agents is a revolutionary approach to the treatment of immunologically mediated diseases. Isolating, propagating, and modifying cells before applying them to patients allows complementation of specific cellular functions, which opens astonishing new possibilities for gain-of-function antigen-specific treatments in autoimmunity, chronic inflammatory disorders, and transplantation. This critical review presents a systematic assessment of the potential clinical risks posed by cell-based immunotherapy, focusing on treatment of renal transplant recipients with regulatory macrophages as a concrete example.

The combination of radiotherapy, temozolomide and valproic acid (VPA) has shown some promise in retrospective analyses of patients with glioblastoma, although their mechanisms of action remain unknown.We investigated the in vitro and in vivo effects of pretreating glioma cells with temozolomide and VPA as an immunization strategy to boost an adaptive immune response in a syngeneic mouse model.Temozolomide and VPA induced autophagy in GL261 glioma cells, and caused tumor antigen-specific T-cells to become activated effector T-cells. Mice with a pre-existing glioma showed no improvement in clinical outcome when immunized with temozolomide- and VPA-treated glioma cells.Although temozolomide and VPA treatment of glioma cells can boost the adaptive immune response, in the context of a vaccine therapy, additional factors are necessary to eradicate the tumor and improve survival.

Microglial cells, the resident macrophage population of the central nervous system (CNS), actively scan tissue under both normal and pathologic contexts. Their resulting engagement can become either neuroprotective or neurotoxic, leading to amelioration or aggravation of disease progression. In this review, we focus on the molecular signaling molecules involved in microglial responses and discuss observations demonstrating the diverse effects of microglia in animal models of CNS diseases.

Focal cerebral ischemia and traumatic brain injury induce an escalating amount of cell death because of harmful mediators diffusing from the original lesion site. Evidence suggests that healthy cells surrounding these lesions attempt to protect themselves by producing endocannabinoids (eCBs) and activating cannabinoid receptors, the molecular target for marijuana-derived compounds. Indeed, activation of cannabinoid receptors reduces the production and diffusion of harmful mediators. Here, we provide evidence that an exception to this pattern is found in experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis. We show that cell damage induced by EAE does not lead to increase in eCBs, even though cannabinoid receptors are functional because synthetic cannabinoid agonists are known to confine EAE-induced lesions. This lack of eCB increase is likely due to IFN-γ, which is released by primed T cells invading the CNS. We show that IFN-γ disrupts the functionality of purinergic P2X 7 receptors, a key step controlling eCB production by microglia, the main source of eCBs in brain. Accordingly, induction of EAE in P2X 7 −/− mice results in even lower eCB levels and more pronounced cell damage than in wild-type mice. Our data suggest that the high level of CNS IFN-γ associated with EAE disrupts eCB-mediated neuroprotection while maintaining functional cannabinoid receptors, thus providing additional support for the use of cannabinoid-based medicine to treat multiple sclerosis.

The cytoplasm of neural cells contain millimolar amounts of ATP, which flood the extracellular space after injury, activating purinergic receptors expressed by glial cells and increasing gliotransmitter production. These gliotransmitters, which are thought to orchestrate neuroinflammation, remain widely uncharacterized. Recently, we showed that microglial cells produce 2-arachidonoylglycerol (2-AG), an endocannabinoid known to prevent the propagation of harmful neuroinflammation, and that ATP increases this production by threefold at 2.5 min (Witting et al., 2004). Here we show that ATP increases 2-AG production from mouse astrocytes in culture, a response that is more rapid (i.e., significant within 10 sec) and pronounced (i.e., 60-fold increase at 2.5 min) than any stimulus-induced increase in endocannabinoid production reported thus far. Increased 2-AG production from astrocytes requires millimolar amounts of ATP, activation of purinergic P2X7receptors, sustained rise in intracellular calcium, and diacylglycerol lipase activity. Furthermore, we show that astrocytes express monoacylglycerol lipase (MGL), the main hydrolyzing enzyme of 2-AG, the pharmacological inhibition of which potentiates the ATP-induced 2-AG production (up to 113-fold of basal 2-AG production at 2.5 min). Our results show that ATP greatly increases, and MGL limits, 2-AG production from astrocytes. We propose that 2-AG may function as a gliotransmitter, with MGL inhibitors potentiating this production and possibly restraining the propagation of harmful neuroinflammation.

Growing evidence suggests that a major physiological function of the cannabinoid signaling system is to modulate neuroinflammation. This review discusses the anti-inflammatory properties of cannabinoid compounds at molecular, cellular and whole animal levels, first by examining the evidence for anti-inflammatory effects of cannabinoids obtained using in vivo animal models of clinical neuroinflammatory conditions, specifically rodent models of multiple sclerosis, and second by describing the endogenous cannabinoid (endocannabinoid) system components in immune cells. Our aim is to identify immune functions modulated by cannabinoids that could account for their anti-inflammatory effects in these animal models.

Astrocytes play an important role in neuroprotective responses. Recent studies indicate that endothelin-1, a neuropeptide upregulated during brain injury, increases levels of the endocannabinoid anandamide, a lipid with neuroprotective properties, in astrocytes in primary cultures. However, whether this neuropeptide also alters levels of 2-arachidonoyl glycerol (2-AG), the most abundant endocannabinoid in the CNS, in astrocytes remains unknown. In addition, 2-AG levels in astrocytes have never been measured. In this report we use chemical ionization gas chromatography/mass spectrometry to quantify picomole amounts of 2-AG in primary cultures of mouse astrocytes. We also demonstrate that endothelin-1 increases 2-AG production by 5-fold in these cells, a response that requires extracellular calcium and endothelin-1(A) receptor engagement. Immunocytochemistry showed that although cultured mouse neurons and microglia express cannabinoid receptors, cultured astrocytes do not. The data suggest that endothelin-1 modulates 2-AG production in astrocytes and that this endocannabinoid may participate in paracrine signaling toward neurons and microglia.

During neuroinflammation, activated microglial cells migrate toward dying neurons, where they exacerbate local cell damage. The signaling molecules that trigger microglial cell migration are poorly understood. In this paper, we show that pathological overstimulation of neurons by glutamate plus carbachol dramatically increases the production of the endocannabinoid 2-arachidonylglycerol (2-AG) but only slightly increases the production of anandamide and does not affect the production of two putative endocannabinoids, homo-gamma-linolenylethanolamide and docosatetraenylethanolamide. We further show that pathological stimulation of microglial cells with ATP also increases the production of 2-AG without affecting the amount of other endocannabinoids. Using a Boyden chamber assay, we provide evidence that 2-AG triggers microglial cell migration. This effect of 2-AG occurs through CB2 and abnormal-cannabidiol-sensitive receptors, with subsequent activation of the extracellular signal-regulated kinase 1/2 signal transduction pathway. It is important to note that cannabinol and cannabidiol, two nonpsychotropic ingredients present in the marijuana plant, prevent the 2-AG-induced cell migration by antagonizing the CB2 and abnormal-cannabidiol-sensitive receptors, respectively. Finally, we show that microglial cells express CB2 receptors at the leading edge of lamellipodia, which is consistent with the involvement of microglial cells in cell migration. Our study identifies a cannabinoid signaling system regulating microglial cell migration. Because this signaling system is likely to be involved in recruiting microglial cells toward dying neurons, we propose that cannabinol and cannabidiol are promising nonpsychotropic therapeutics to prevent the recruitment of these cells at neuroinflammatory lesion sites.

Administering immunoregulatory cells to patients as medicinal agents is a potentially revolutionary approach to the treatment of immunologically mediated diseases. Presently, there are no satisfactory, clinically applicable methods of tracking human cells in patients with adequate spatial resolution and target cell specificity over a sufficient period of time. Laser ablation–inductively coupled plasma mass spectrometry (LA-ICP-MS) represents a potential solution to the problem of detecting very rare cells in tissues. In this article, this exquisitely sensitive technique is applied to the tracking of gold-labeled human regulatory macrophages (Mregs) in immunodeficient mice. Optimal conditions for labeling Mregs with 50-nm gold particles were investigated by exposing Mregs in culture to variable concentrations of label: Mregs incubated with 3.5 × 109 particles/ml for 1 h incorporated an average of 3.39 × 108 Au atoms/cell without loss of cell viability. Analysis of single, gold-labeled Mregs by LA-ICP-MS registered an average of 1.9 × 105 counts/cell. Under these conditions, 100% labeling efficiency was achieved, and label was retained by Mregs for ≥36 h. Gold-labeled Mregs adhered to glass surfaces; after 24 h of culture, it was possible to colabel these cells with human-specific 154Sm-tagged anti–HLA-DR or 174Yb-tagged anti-CD45 mAbs. Following injection into immunodeficient mice, signals from gold-labeled human Mregs could be detected in mouse lung, liver, and spleen for at least 7 d by solution-based inductively coupled plasma mass spectrometry and LA-ICP-MS. These promising results indicate that LA-ICP-MS tissue imaging has great potential as an analytical technique in immunology.

Introduction: Diabetes is a disease affecting millions of people worldwide, and is a major independent risk factor for cardiovascular disease (CVD). Despite a vast amount of research, the molecular mechanisms that link diabetes to CVD are not well understood. Current evidence suggests that increased flux through the hexosamine biosynthetic pathway (HBP) contributes to the development of hyperglycemia-associated diabetic complications. Our data suggest that increased HBP flux can induce vascular ER stress and accelerate atherogenesis in a mouse model. We hypothesized that this process can be attenuated by inhibiting the first and rate-limiting enzyme in the HBP - glutamine fructose-6-phosphate amidotransferase (GFAT) - using small molecules. Methods and Results: Using high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS) we have developed a methodology to monitor and quantify the levels of the end product of the HBP, uridine diphosphate N -acetylglucosamine (UDP-GlcNAc). Treatment of HepG2 cells with glucosamine (0.2-5 mM), or adenovirus-directed overexpression of GFAT caused a 3-7 fold increase in UDP-GlcNAc ( P< 0.001 & P< 0.05, respectively). Inhibition of GFAT with three novel compounds - amrinone, lapachol or alloxan - decreased levels of UDP-GlcNAc by 1.5 ( P< 0.05), 3 ( P< 0.05) and 3.5-fold ( P< 0.001), respectively. Furthermore, we show that by modulating HBP flux, we can regulate ER stress levels in cultured HepG2 cells. The physiological relevance of this mechanism is supported by evidence of HBP augmentation in a hyperglycemic mouse model. Conclusions: These results support a role for the HBP in the development of atherosclerosis. Currently, MALDI imaging mass spectrometry is being performed on tissue sections to compare the levels of UDP-GlcNAc directly in hyperglycemic vs. normoglycemic mice. These studies may lead to the identification and validation of novel targets for the development of new pharmaceuticals to prevent diabetic atherosclerosis.

Administering immunoregulatory cells as medicinal agents is a revolutionary approach to the treatment of immunologically mediated diseases. Isolating, propagating, and modifying cells before applying them to patients allows complementation of specific cellular functions, which opens astonishing new possibilities for gain-of-function antigen-specific treatments in autoimmunity, chronic inflammatory disorders, and transplantation. This critical review presents a systematic assessment of the potential clinical risks posed by cell-based immunotherapy, focusing on treatment of renal transplant recipients with regulatory macrophages as a concrete example.

Hospitals and health systems should take the following steps to improve their revenue cycle performance: Collect patient responsibility amounts up front. Reduce credit balance accounts. Reduce preregistered patient no-shows. Identify and manage unbilled accounts receivable.

Anna Triandafyllidou and Thanos Maroukis, Migrant smuggling: irregular migration from Asia and Africa to Europe, New York: Palgrave Macmillan, 2012, 256 pp. (ISBN: 9780230296374)

Headlines like this have featured European newspapers in recent times. What has emerged is that migration is often connected to illicit activities of migrant smugglers. This has called for a fight against irregular migration.

The CNS is considered immune privileged due to the blood-brain barrier and the absence of conventional lymphatics. Nonetheless, T cell immune responses specific for CNS Ag have been documented. Where these events are initiated and what cellular mechanisms are involved remain unknown. In this study, we established an experimental mouse model to evaluate the requirements for priming CD8+ T cells following the cross-presentation of intracranial Ag. Surprisingly, we find that even with a damaged blood-brain barrier, Ag presentation occurs in regional lymph nodes and not within the CNS itself. Only once the responding cells have expanded can they traffic to the site of CNS injury. Cross-presentation of intracranial Ag is efficient and the subsequent priming of CD8+ T cells is dependent on CD4+ T cell help and CD40 signaling in host APCs. Our findings have important implications for the initiation of T cell immune responses toward CNS Ags.

Endogenous cannabinoid ligands (endocannabinoids) produced by neurons, astrocytes, and microglial cells activate cannabinoid receptors, the molecular target for marijuana's bioactive ingredient Δ 9 -tetrahydrocannabinol. The molecular mechanism underlying the production of the most abundant endocannabinoid, 2-arachidonoylglycerol (2-AG), is unclear. A prevalent hypothesis proposes that activation of metabotropic receptors coupled to the phosphatidylinositol-specific phospholipase C and diacylglycerol (DG) lipase pathway will systematically lead to increases in 2-AG production. Here, we show that ATP increases 2-AG production by cultured microglial cells in a phosphatidylinositol-specific phospholipase C and DG lipase-dependent manner. However, efficacious activation of metabotropic P2Y purinergic receptors coupled to phosphatidylinositol-specific phospholipase C does not increase 2-AG production. This suggests that ionotropic, and not metabotropic, purinergic receptors control 2-AG production at an unexpected enzymatic step of its metabolic pathway. We show that activation of P2X 7 ionotropic receptors, which are highly permeable to calcium, is necessary and sufficient to increase 2-AG production in microglial cells. We also show that the sustained rise in intracellular calcium induced by activation of P2X 7 receptors directly increases DG lipase activity while inhibiting the activity of monoacylglycerol lipase, the enzyme that degrades 2-AG. This inverse sensitivity of DG lipase and monoacylglycerol lipase to calcium constitutes an original and efficient modality for sustained accumulation of 2-AG. Because prolonged increases in 2-AG amounts in brain parenchyma are thought to orchestrate neuroinflammation, the enzymatic steps involved in 2-AG synthesis and degradation by microglial cells constitute appealing targets for therapy aimed at controlling exacerbated neuroinflammation.

Focal cerebral ischemia (FCI) induces rapid neuronal death in the ischemic core, which gradually expands toward the penumbra, partly as the result of a neuroinflammatory response. It is known that propagation of neuroinflammation involves microglial cells, the resident macrophages of the brain, which are highly motile when activated by specific signals. However, the signals that increase microglial cell motility in response to FCI remain mostly elusive.Here, we tested the hypothesis that endocannabinoids mediate neuroinflammation propagation by increasing microglial cell motility. We found that, in mouse cerebral cortex, FCI greatly increases palmitoylethanolamide (PEA), only moderately increases anandamide [arachidonylethanolamide (AEA)], and does not affect 2-arachidonoylglycerol levels. We also found that PEA potentiates AEA-induced microglial cell migration, without affecting other steps of microglial activation, such as proliferation, particle engulfment, and nitric oxide production. This potentiation of microglial cell migration by PEA involves reduction in cAMP levels. In line with this, we provide evidence that PEA acts through Gi/o-coupled receptors. Interestingly, these receptors engaged by PEA are pharmacologically distinct from CB1and CB2cannabinoid receptors, as well as from the WIN and abn-CBD (abnormal-cannabidiol) receptors, two recently identified cannabinoid receptors.Our results show that PEA and AEA increase after FCI and synergistically enhance microglial cell motility. Because such a response could participate in the propagation of the FCI-induced neuroinflammation within the CNS, and because PEA is likely to act through its own receptor, a better understanding of the receptor engaged by PEA may help guide the search for improved therapies against neuroinflammation.