A metapopulation approach to identify targets for Wolbachia-based dengue control.

Over the last decade, the release of Wolbachia-infected Aedes aegypti into the natural habitat of this mosquito species has become the most sustainable and long-lasting technique to prevent and control vector-borne diseases, such as dengue, zika, or chikungunya. However, the limited resources to generate such mosquitoes and their effective distribution in large areas dominated by the Aedes aegypti vector represent a challenge for policymakers. Here, we introduce a mathematical framework for the spread of dengue in which competition between wild and Wolbachia-infected mosquitoes, the cross-contagion patterns between humans and vectors, the heterogeneous distribution of the human population in different areas, and the mobility flows between them are combined. Our framework allows us to identify the most effective areas for the release of Wolbachia-infected mosquitoes to achieve a large decrease in the global dengue prevalence.

Epigenética en obesidad y diabetes tipo 2: papel de la nutrición, limitaciones y futuras aplicaciones / Epigenetics in obesity and type 2 diabetes: role of nutrition, limitations and future applications

In the last years, epigenetics is helping to explain the mechanisms non dependent on the genetic sequence by which the nutrients and other environmental factors contribute to modulate gene expression and disease development. Obesity and type 2 diabetes are two diseases that have been linked to changes in epigenetic marks (particularly DNA methylation, covalent modifications of histones and miRNAs). These epigenetic changes appear to be influenced, mainly in the perinatal period but also in adulthood, by environmental factors such as hyperglycemia or the diet. Among the food compounds that have been linked to epigenetic modifications, there are methyl donor groups, excessive or deficient caloric intake, short chain fatty acids, some minerals and antioxidant vitamins, and various compounds of plant origin, as catechins, isoflavones or isothiocyanates. EWAS studies, that analyze the methylation of thousands of CpG sites in thousands of individuals, will contribute in the next years to identify some of the epigenetic marks that can be used as early predictors of metabolic risk, as well as some intimate mechanisms that explain the development of obesity, type 2 diabetes and its complications. Moreover, epigenetic marks, among them the CpG-SNPs, can be heritable but some of them could be potentially reversible. One of the medium-term objectives is to develop drug or diet-related treatments that could delay or even reverse these epigenetic changes.

GABAergic and cholinergic neurons exhibit high-affinity nerve growth factor binding in rat basal forebrain.

We have used dissociated, rat basal forebrain cultures to identify specific cell types that are potentially responsive to nerve growth factor (NGF). Expression of high-affinity NGF binding sites was examined. A subpopulation of cells containing choline acetyltransferase (CAT), the acetylcholine-synthesizing enzyme, exhibited high-affinity binding, employing combined immunocytochemistry and 125I-NGF radioautography. Unexpectedly, a gamma-aminobutyric acid (GABA)-containing cell group also expressed high-affinity binding. These cells that exhibit high-affinity binding appear to be neurons since they stain positively with the neuron marker, neuron-specific enolase, and negatively with the nonneuron marker, glial fibrillary acidic protein. Our observations suggest that NGF may regulate multiple brain systems and functions that have yet to be explored. Conversely, only subsets of cholinergic or GABA neurons expressed high-affinity binding, suggesting that these transmitter populations are composed of differentially responsive subpopulations.

Differential expression of the nerve growth factor receptor gene in multiple brain areas.

Recent work has indicated that the trophic protein, nerve growth factor (NGF), is detectable in several brain regions, in addition to its well-known localization to the periphery. In addition, a number of cholinergic populations in the brain respond to NGF by increasing enzymes involved in acetylcholine metabolism. It is well recognized that responsiveness to NGF is dependent on expression of specific receptors; we have recently detected expression by the responsive rat basal forebrain/septal, cholinergic neurons, suggesting that NGF plays a physiologic role in the development of this brain pathway. To define a potential role for NGF in other rat brain regions, we isolated a rat receptor cDNA clone to use as a probe to detect receptor message by sensitive Sl nuclease protection experiments. Our studies indicate that the NGF receptor (NGF-R) gene is expressed by anatomically, functionally and biochemically diverse populations, widely distributed in the rat brain, and is not restricted to the basal forebrain/septal region. We detect NGF receptor message in frontal cortex, hippocampus, caudate, cerebellum and olfactory bulb. Moreover, developmental profiles of steady-state quantities varied differently for each area. Our observations support the contention that NGF regulates multiple brain systems, in addition to forebrain cholinergic pathways.

Localization of high-affinity and low-affinity nerve growth factor receptors in cultured rat basal forebrain.

Previous work has indicated that nerve growth factor specifically and selectively increases choline acetyltransferase and acetylcholinesterase in organotypic cultures of rat basal forebrain-medial septal area. To determine whether these actions are potentially receptor-mediated, organotypic and dissociated basal forebrain-medial septal area cultures were examined. Two independent methods, [125I]nerve growth factor binding and immunocytochemistry with a monoclonal nerve growth factor receptor antibody (192-IgG), detected specific receptors. The nerve growth factor receptors were localized to two different cellular populations: flat, large, non-neuron-like cells, and small, round, process-bearing, neuron-like cells. Dissociation studies with [125I]nerve growth factor suggested that high-affinity receptors were localized to the neuron-like population, while only low-affinity receptors were localized to the non-neuron-like cells. We tentatively conclude that nerve growth factor may elicit cholinergic effects by directly binding to high-affinity receptors on neurons. To begin examining receptor regulation, cultures were exposed to exogenous, unlabeled nerve growth factor continuously for 10 days before binding studies were performed. Prior exposure to nerve growth factor did not alter binding characteristics of the receptor, using the present methods.

Nerve growth factor selectively increases cholinergic markers but not neuropeptides in rat basal forebrain in culture.

We have previously used organotypic cultures to study mechanisms regulating phenotypic expression of neurotransmitter characters in the brain. Our previous work indicated that nerve growth factor (NGF) specifically increased the activity of choline acetyltransferase (CAT) in striatal cholinergic interneurons. In the present study we examined the effect of NGF on neurons of fetal rat basal forebrain-medial septal area (BF-MS) maintained in organotypic culture. Treatment with 200 biological units/ml of NGF resulted in a 3- to 6-fold increase in the specific activity of CAT. This effect was specifically blocked by anti-NGF antiserum, whereas treatment with antiserum alone did not alter the cholinergic enzyme. NGF also elicited a marked increase in CAT staining intensity, using a monoclonal antibody directed against the enzyme. Further, the number of CAT-positive neurons appeared to increase in the NGF-treated cultures. Exposure to NGF also increased the activity of another cholinergic marker, the catabolic enzyme, acetylcholinesterase. The effect of NGF appeared to be highly selective, since substance P and somatostatin levels were unchanged by NGF treatment.

Developmentally regulated expression of the nerve growth factor receptor gene in the periphery and brain.

Nerve growth factor (NGF) regulates development and maintenance of function of peripheral sympathetic and sensory neurons. A potential role for the trophic factor in brain has been detected only recently. The ability of a cell to respond to NGF is due, in part, to expression of specific receptors on the cell surface. To study tissue-specific expression of the NGF receptor gene, we have used sensitive cRNA probes for detection of NGF receptor mRNA. Our studies indicate that the receptor gene is selectively and specifically expressed in sympathetic (superior cervical) and sensory (dorsal root) ganglia in the periphery, and by the septum-basal forebrain centrally, in the neonatal rat in vivo. Moreover, examination of tissues from neonatal and adult rats reveals a marked reduction in steady-state NGF receptor mRNA levels in sensory ganglia. In contrast, a 2- to 4-fold increase was observed in the basal forebrain and in the sympathetic ganglia over the same time period. Our observations suggest that NGF receptor mRNA expression is developmentally regulated in specific areas of the nervous system in a differential fashion.

Nerve growth factor promotes cholinergic development in brain striatal cultures.

We have examined the effect of the trophic protein, nerve growth factor (NGF), on organotypic cultures of fetal rat striatum. Treatment of cultures with NGF for 10-11 days resulted in a 5- to 12-fold increase in the specific activity of the cholinergic enzyme choline acetyltransferase (CAT; EC 2.3.1.6). in a dose-dependent fashion. This effect was not elicited by insulin, ferritin, or cytochrome c, proteins similar in structure or physicochemical properties to NGF. The effect of NGF on CAT activity was specifically blocked by anti-NGF antiserum, whereas treatment with the antiserum alone did not have a significant effect on the enzyme. Immunocytochemical studies of the treated cultures, using a monoclonal antibody directed against CAT, revealed positively stained neurons exhibiting dendritic and axonal processes. NGF did not have an effect on total protein content of the striatal cultures, suggesting a highly specific effect. Moreover, levels of substance P, a peptide localized to other, noncholinergic neurons, were not altered by NGF. Substance P remained unchanged after treatment with NGF for 12 days, whereas CAT activity increased 12-fold in sister cultures. Although the mechanisms of action of NGF on striatal cholinergic interneurons remain to be determined, the marked, specific response of CAT suggests that this well-defined trophic protein may play a critical role in normal brain development.