ANHIDRASA CARBÓNICA DE Plasmodium falciparum: UN BLANCO ÚTIL PARA EL DISEÑO DE MEDICAMENTOS ANTIMALÁRICOS Y COMPUESTOS BLOQUEADORES DE LA TRANSMISIÓN DE MALARIA CARBONIC ANHYDRASE IN Plasmodium falciparum: A USEFUL TARGET FOR ANTIMALARIAL DRUG DESIGNING AND MALARIA BLOCKING TRANSMISSION COMPOUNDS

La anhidrasa carbónica es una metaloenzima que cataliza la conversión reversible del CO2 a bicarbonato, un componente metabólico indispensable para la síntesis de pirimidinas de novo por Plasmodium spp y los procesos de exflagelación llevados a cabo por el parásito al interior del mosquito vector. La enzima participa además en el transporte del bicarbonato dentro y fuera de las células para evitar un desequilibrio en el sistema CO2/HCO3- y la alteración del pH al interior de las células y en el espacio intercelular. Por lo tanto, al inhibir la enzima, ya sea en el parásito o en el insecto vector, se podría conducir a una disminución de la replicación y al detrimento y/o muerte del parásito. De esta forma, los inhibidores de anhidrasa carbónica constituyen una alternativa, tanto terapéutica como de bloqueo de la transmisión, para el control de la malaria. La actividad anti-Plasmodium in vitro de algunos compuestos inhibidores de anhidrasa carbónica ya se ha determinado. Sin embargo, la eficacia in vivo y el mecanismo por el cual los inhibidores son capaces de afectar el desarrollo del parásito en los mosquitos vectores permanecen aún por evaluarse. En el marco del proyecto de investigación "Evaluación de inhibidores de anhidrasa carbónica como medidas terapéuticas y de bloqueo de la transmisión de malaria" este artículo presenta una revisión del estado del arte sobre el papel de la anhidrasa carbónica de Plasmodium spp y el uso de inhibidores específicos de esta enzima como una estrategia para el tratamiento de la malaria y el bloqueo de la transmisión de la enfermedad. Se incluyeron artículos publicados en los últimos 59 años, identificados a partir de la bases de datos bibliográficos PubMed y ScienceDirect, cruzando las palabras claves, al igual que artículos recopilados por los autores y se analizan e integran los resultados de investigaciones publicadas alrededor del tema.Carbonic anhydrase is a metalloenzyme that catalyzes the reversible conversion of CO2 to bicarbonate, an essential metabolic component used by the malaria parasites for de novo synthesis of pyrimidines and the exflagelation of gametocytes inside the mosquito vector. Carbonic anhydrase is involved in the transport of bicarbonate. This enzyme participates in transport of bicarbonate inside and outside the cells to avoid an imbalance in the system CO2/HCO3- and alteration of pH in the interior of the cell as well as in the intercellular space. Therefore, inhibition of this enzyme either in the parasite or the insect vector, could lead to a decrease in replication and to the detriment and/or death of the parasite. Given the importance of carbonic anhydrase in the metabolism, development and survival of Plasmodium, it could be postulated that carbonic anhydrase inhibitors are both a therapeutic and a blocking transmission alternative. Previous studies have demonstrated the in vitro anti-Plasmodium activity of some inhibitors. However, it is necessary to determine their effectiveness to confirm its usefulness in the treatment or blocking malaria transmission and the mechanism by which these inhibitors are able to affect the development of the parasite in the mosquito vector. In this paper we present a review about the role of carbonic anhydrase in Plasmodium spp and using some specific inhibitors as a strategy for malaria treatment and transmission blocking strategy. Articles published in the past 59 years identified from bibliographic database (PubMed and ScienceDirect) and papers collected by the authors were included

New model of action for mood stabilizers: phosphoproteome from rat pre-frontal cortex synaptoneurosomal preparations.

BACKGROUND: Mitochondrial short and long-range movements are necessary to generate the energy needed for synaptic signaling and plasticity. Therefore, an effective mechanism to transport and anchor mitochondria to pre- and post-synaptic terminals is as important as functional mitochondria in neuronal firing. Mitochondrial movement range is regulated by phosphorylation of cytoskeletal and motor proteins in addition to changes in mitochondrial membrane potential. Movement direction is regulated by serotonin and dopamine levels. However, data on mitochondrial movement defects and their involvement in defective signaling and neuroplasticity in relationship with mood disorders is scarce. We have previously reported the effects of lithium, valproate and a new antipsychotic, paliperidone on protein expression levels at the synaptic level. HYPOTHESIS: Mitochondrial function defects have recently been implicated in schizophrenia and bipolar disorder. We postulate that mood stabilizer treatment has a profound effect on mitochondrial function, synaptic plasticity, mitochondrial migration and direction of movement. METHODS: Synaptoneurosomal preparations from rat pre-frontal cortex were obtained after 28 daily intraperitoneal injections of lithium, valproate and paliperidone. Phosphorylated proteins were identified using 2D-DIGE and nano LC-ESI tandem mass spectrometry. RESULTS: Lithium, valproate and paliperidone had a substantial and common effect on the phosphorylation state of specific actin, tubulin and myosin isoforms as well as other proteins associated with neurofilaments. Furthermore, different subunits from complex III and V of the electron transfer chain were heavily phosphorylated by treatment with these drugs indicating selective phosphorylation. CONCLUSIONS: Mood stabilizers have an effect on mitochondrial function, mitochondrial movement and the direction of this movement. The implications of these findings will contribute to novel insights regarding clinical treatment and the mode of action of these drugs

Actin and tubulin were phosphorylated after mood stabilizer treatment.

<p>A. 2D-DIGE gel image stained with Sypro Ruby stain showing phosphorylation of synaptoneurosomal preparations of saline treated animals (controls). B. location of actin identified spots (green spot) and tubulin (red spot) in saline treated preparations. C. Spots identified with actin antibodies during Western blots (circled) of synaptoneurosomal preparations after mood stabilizer treatment. D. Western blot identification of phosphorylated actin (circled). E. 2D-DIGE gel image showing phosphorylation after mood stabilizer treatment and the location of tubulin identified spots as indicated by circle. F. Spots identified with tubulin antibodies (circled).</p

Mitochondrial staining with MitoTracker® green (total mitochondria) and MitoTracker red® (active function mitochondria) shows different morphology in cells at a given time indicative of rapid changes in movement and mitochondrial function as exemplified by the saline controls (A, B, C and D).

<p>Anterograde mitochondrial movement (away from the cell nuclei) was observed after treatment with 20 µM lithium (E) and 50 µM Paliperidone (F) as indicated by long filamentous mitochondria. Ballooning and concentration around cell nuclei (blue) was observed after treatment with 50 µM Clozapine (G). Detrimental effects were observed in mitochondria after treatment with 50 µM Haloperidol (H). Superimposed images shown in yellow.</p

Protein phosphorylation in synaptoneurosomal preps from rat PFC as a result of chronic lithium, valproate, and paliperidone treatment.

<p>All proteins shown in this table exhibited a protein identification probability of 100% and were chosen based on the following criteria: minimum protein volume was set at 200 and only proteins with a 2 fold or more difference in protein expression, a 100% presence in all gel images, and P-values<0.05 (ANOVA) were selected. (%SC = percent sequence coverage, N = number of unique peptides).</p

Mood stabilizer treatment resulted in increased mitochondrial transport to the synapse in cell culture as indicated by increased staining of neuronal processes and rat PFC tissue.

<p>From left to right: MitoTracker (red), SYN1 (green), and superimposed images (red and green). A. Saline control, B. Lithium (1 mM) treated cells, C. Paliperidone (10 µM) treated cells, D. Saline treated rat PFC tissue slice (left to right: MitoTracker red, DAPI nuclear stain and superimposed image), E. Lithium (22 mg/Kg) treated rat PFC tissue slice (left to right: MitoTracker red, DAPI and superimposed image). F. A close-up of localization of Mito Tracker red (red, left) and SYN1 (green) in paliperidone (0.1 µM) treated cells. Nuclei shown in blue (Secondary antibody: Alexafluor488 Mouse 1∶1000, Mitotracker CFX-Ros 50 nM, Hoecsht nuclear stain 1∶1000).</p

Deployment of innovative genetic vector control strategies: progress on regulatory and biosafety aspects, capacity building and development of best-practice guidance

In the ongoing fight against vectors of human diseases, disease endemic countries (DECs) may soon benefit from innovative control strategies involving modified insect vectors. For instance, three promising methods (viz. RIDL® [Release of Insects with a Dominant Lethal], Wolbachia infection, and refractory mosquito technology) are being developed by researchers around the world to combat Aedes aegypti, the primary mosquito vector of viral fevers such as dengue (serotypes 1-4), chikungunya and yellow fever. Some of these techniques are already being extended to other vectors such as Aedes albopictus (the secondary vector of these diseases) and Anopheles mosquito species that transmit malaria. To enable DECs to take advantage of these promising methods, initiatives are underway that relate to biosafety, risk assessment and management, and ethical-social-cultural (ESC) aspects to consider prior to and during the possible deployment of these technologies as part of an integrated vector control programme. This is a brief overview of the objectives and timelines of some of the initiatives being championed by international institutions, including the United Nations Development Programme (UNDP), the World Health Organization (WHO) and the Grand Challenges in Global Health (GCGH) initiative co-sponsored by the Bill &amp; Melinda Gates Foundation