Urinary metabolites and antioxidant products of exogenous melatonin in the mouse

Exogenous melatonin is widely used for sleep disorders and has potential value in neuroprotection, cardioprotection and as an antioxidant. Here, a novel method is described for the determination of melatonin and six metabolites in mouse urine by use of LC-MS/MS and GC-MS. LC-MS/MS is used for the measurement of melatonin, N1-acetyl-5-methoxykynuramine (AMK), N1-acetyl-N2-formyl-5-methoxykynuramine (AFMK) and 6-hydroxymelatonin (6-HMEL), while GC/MS is used for the determination of N-[2-(5-methoxy-2-oxo-2,3-dihydro-1H-indol-3-yl)-ethyl]-acetamide (2-OMEL) and cyclic 3-hydroxymelatonin (3-HMEL) with detection limits on column of 0.02-0.5 pmol, depending on the metabolite. Following oral administration of melatonin to mice, a 0-24 hr urine collection revealed the presence of melatonin (0.2% dose), 6-HMEL (37.1%) and NAS (3.1%) comprising >90% of the total metabolites; AMK and AFMK were also detected at 0.01% each; 2-OMEL was found at 2.2% of the dose, which is >100 times more than the AMK/AFMK pathway, and comprises >5% of the melatonin-related material detected in mouse urine. 3-HMEL was largely found as a sulfate conjugate. These studies establish sensitive assays for determination of six melatonin metabolites in mouse urine and confirm the potential for antioxidant activity of melatonin through the identification in vivo of AMK and AFMK, ring-opened metabolites with a high capacity for scavenging reactive oxygen species.

Valorization of agrobiodiversity products and strengthening of local identities in the Peruvian Andes: Experiences from the BioAndes Programme

In the Peruvian Andes, a long history of interaction between the local populations and their natural environment has led to extraordinary levels of agrobiodiversity. However, in sharp contrast with this biological wealth, Andean indigenous populations live under most precarious conditions. Moreover, natural resources are undergoing severe degradation processes and local knowledge about biodiversity management is under serious pressure. Against this background, the BioAndes Programme is developing initiatives based on a biocultural approach that aim at fostering biodiversity through the enhancement of cultural processes. On the basis of intercultural dialogue, joint learning and capacity development, and transdisciplinary action-research, indigenous communities, development practitioners, and researchers strive for the creation of innovative ways to contribute to more sustainable economic, socio-cultural, and political valorization of Andean biodiversity. Project activities are diverse and range from the cultivation, transformation, and commercialization of organic Andean fruits in San Marcos, Cajamarca Department, to the recuperation of natural dying techniques for alpaca wool and traditional weaving in Pitumarca, Cusco Department, and the promotion of responsible ecotourism in both regions. Based on the projects’ first two-years of experience, the following lessons learnt will be presented and discussed: 1. The economic valorization and commercialization of local products can be a powerful tool for the revival and innovation of eroded know-how; at the same time it contributes to the strengthening of local identities, in parallel with the empowerment of marginalized groups such as smallholders and women. 2. Such initiatives are only successful when they are embedded within activities that go beyond the focus on local products and seek the valorization of the entire natural and cultural landscape (e.g. through the promotion of agrotourism and local gastronomy, more sustainable management of local resources including the restoration of ecosystems, and the realization of inventories of local agrobiodiversity and the knowledge related to it). 3. The sustainability of these initiatives, which are often externally induced, is conditioned by the ability of local actors to acquire ownership of projects and access to the knowledge required to carry them out, which also means developing the personal and institutional capacities for handling the whole chain from production to commercialization. 4. The confrontation of different economic rationalities and their underlying worldviews that occur when local or indigenous people integrate into the market economy implies the need for a dialogical co-production of knowledge and collective action by local people, experts from NGOs, and political authorities in order to better control the conditions relating to the market economy. The valorization of local agrobiodiversity shows much potential for enhancing natural and cultural diversity in Southern countries, but only when local communities can participate in the shaping of the conditions under which this happens. Such activities should be designed in the mid- to long-term as part of social learning processes that are carefully embedded in the local context. Supporting institutions play a crucial role in these processes, but should see themselves only as facilitators, while ensuring that control and ownership remain with the local actors.

Toxoplasma gondii Infection in Kyrgyzstan: Seroprevalence, Risk Factor Analysis, and Estimate of Congenital and AIDS-Related Toxoplasmosis

Background HIV-prevalence, as well as incidence of zoonotic parasitic diseases like cystic echinococcosis, has increased in the Kyrgyz Republic due to fundamental socio-economic changes after the breakdown of the Soviet Union. The possible impact on morbidity and mortality caused by Toxoplasma gondii infection in congenital toxoplasmosis or as an opportunistic infection in the emerging AIDS pandemic has not been reported from Kyrgyzstan. Methodology/Principal Findings We screened 1,061 rural and 899 urban people to determine the seroprevalence of T. gondii infection in 2 representative but epidemiologically distinct populations in Kyrgyzstan. The rural population was from a typical agricultural district where sheep husbandry is a major occupation. The urban population was selected in collaboration with several diagnostic laboratories in Bishkek, the largest city in Kyrgyzstan. We designed a questionnaire that was used on all rural subjects so a risk-factor analysis could be undertaken. The samples from the urban population were anonymous and only data with regard to age and gender was available. Estimates of putative cases of congenital and AIDS-related toxoplasmosis in the whole country were made from the results of the serology. Specific antibodies (IgG) against Triton X-100 extracted antigens of T. gondii tachyzoites from in vitro cultures were determined by ELISA. Overall seroprevalence of infection with T. gondii in people living in rural vs. urban areas was 6.2% (95%CI: 4.8–7.8) (adjusted seroprevalence based on census figures 5.1%, 95% CI 3.9–6.5), and 19.0% (95%CI: 16.5–21.7) (adjusted 16.4%, 95% CI 14.1–19.3), respectively, without significant gender-specific differences. The seroprevalence increased with age. Independently low social status increased the risk of Toxoplasma seropositivity while increasing numbers of sheep owned decreased the risk of seropositivity. Water supply, consumption of unpasteurized milk products or undercooked meat, as well as cat ownership, had no significant influence on the risk for seropositivity. Conclusions We present a first seroprevalence analysis for human T. gondii infection in the Kyrgyz Republic. Based on these data we estimate that 173 (95% CI 136–216) Kyrgyz children will be born annually to mothers who seroconverted to toxoplasmosis during pregnancy. In addition, between 350 and 1,000 HIV-infected persons are currently estimated to be seropositive for toxoplasmosis. Taken together, this suggests a substantial impact of congenital and AIDS-related symptomatic toxoplasmosis on morbidity and mortality in Kyrgyzstan.

Carbon-related Border Adjustment and WTO Law

By expounding the legal foundations of border tax adjustments in international trade regulation, this book lays out the scope and limitations within which border carbon adjustments need to operate. The author examines the extent to which countries can lawfully impose border adjustment measures in relation to the carbon footprint of products on importation and exportation. In doing so, she provides a thorough analysis of the provisions of the WTO Agreement applicable to border carbon adjustments, offers a comprehensive review of relevant case law and engages with the extensive literature on the subject. Given the probability of conflict with non-discrimination rules of the GATT and uncertainty over justification of different designs of carbon-related border adjustment schemes under the exceptions of GATT Article XX, the book argues for a negotiated solution and discusses the possibility of the use of border carbon adjustments under preferential trade agreements.

Structural analogues of the natural products magnolol and honokiol as potent allosteric potentiators of GABA(A) receptors.

Biphenylic compounds related to the natural products magnolol and 4'-O-methylhonokiol were synthesized, evaluated and optimized as positive allosteric modulators (PAMs) of GABA(A) receptors. The most efficacious compounds were the magnolol analog 5-ethyl-5'-hexylbiphenyl-2,2'-diol (45) and the honokiol analogs 4'-methoxy-5-propylbiphenyl-2-ol (61), 5-butyl-4'-methoxybiphenyl-2-ol (62) and 5-hexyl-4'-methoxybiphenyl-2-ol (64), which showed a most powerful potentiation of GABA-induced currents (up to 20-fold at a GABA concentration of 3μM). They were found not to interfere with the allosteric sites occupied by known allosteric modulators, such as benzodiazepines and N-arachidonoylglycerol. These new PAMs will be useful as pharmacological tools and may have therapeutic potential for mono-therapy, or in combination, for example, with GABA(A) receptor agonists.

Phospholipid oxidation generates potent anti-inflammatory lipid mediators that mimic structurally related pro-resolving eicosanoids by activating Nrf2.

Exposure of biological membranes to reactive oxygen species creates a complex mixture of distinct oxidized phospholipid (OxPL) species, which contribute to the development of chronic inflammatory diseases and metabolic disorders. While the ability of OxPL to modulate biological processes is increasingly recognized, the nature of the biologically active OxPL species and the molecular mechanisms underlying their signaling remain largely unknown. We have employed a combination of mass spectrometry, synthetic chemistry, and immunobiology approaches to characterize the OxPL generated from the abundant phospholipid 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine (PAPC) and investigated their bioactivities and signaling pathways in vitro and in vivo. Our study defines epoxycyclopentenones as potent anti-inflammatory lipid mediators that mimic the signaling of endogenous, pro-resolving prostanoids by activating the transcription factor nuclear factor E2-related factor 2 (Nrf2). Using a library of OxPL variants, we identified a synthetic OxPL derivative, which alleviated endotoxin-induced lung injury and inhibited development of pro-inflammatory T helper (Th) 1 cells. These findings provide a molecular basis for the negative regulation of inflammation by lipid peroxidation products and propose a novel class of highly bioactive compounds for the treatment of inflammatory diseases.