Temephos Resistance Status of Aedes aegypti Populations From Havana, Cuba.

Aedes aegypti chemical control remains an indispensable alternative to prevent dengue, Zika, and chikungunya outbreaks in Havana, Cuba. The city of Havana requires constant surveillance because of its bioecological characteristics that favor the proliferation of mosquito vectors of these viruses, which constitutes a high risk to the health of its inhabitants. The goal of this study was to determine the impact of the stopping of temephos applications during the 2 years of the COVID-19 pandemic on the level of susceptibility of Ae. aegypti in 5 municipalities of Havana, Cuba. Larval susceptibility was evaluated by bioassays as described by the World Health Organization. All Ae. aegypti populations tested showed high resistance to temephos. The National Control Program of Ae. aegypti in Cuba will need to promote insecticide rotation policies to prevent the evolution of temephos resistance in Havana.

Improvement and Validation of a Genomic DNA Extraction Method for Human Breastmilk.

The human milk microbiota (HMM) of healthy women can vary substantially, as demonstrated by recent advances in DNA sequencing technology. However, the method used to extract genomic DNA (gDNA) from these samples may impact the observed variations and potentially bias the microbiological reconstruction. Therefore, it is important to use a DNA extraction method that is able to effectively isolate gDNA from a diverse range of microorganisms. In this study, we improved and compared a DNA extraction method for gDNA isolation from human milk (HM) samples to commercial and standard protocols. We evaluated the extracted gDNA using spectrophotometric measurements, gel electrophoresis, and PCR amplifications to assess its quantity, quality, and amplifiability. Additionally, we tested the improved method's ability to isolate amplifiable gDNA from fungi, Gram-positive and Gram-negative bacteria to validate its potential for reconstructing microbiological profiles. The improved DNA extraction method resulted in a higher quality and quantity of the extracted gDNA compared to the commercial and standard protocols and allowed for polymerase chain reaction (PCR) amplification of the V3-V4 regions of the 16S ribosomal gene in all the samples and the ITS-1 region of the fungal 18S ribosomal gene in 95% of the samples. These results suggest that the improved DNA extraction method demonstrates better performance for gDNA extraction from complex samples such as HM.

Effect of Ouabain on Glutamate Transport in the Hippocampus of Rats with LPS-Induced Neuroinflammation.

A lipopolysaccharide (LPS)-induced neuroinflammation rat model was used to study the effects of ouabain (OUA) at low concentrations, which can interact with the Na,K-ATPase, causing the modulation of intracellular signalling pathways in the Central Nervous System. Our study aimed to analyse the effects of OUA on glutamate transport in the hippocampus of rats with LPS-induced neuroinflammation. Adult male Wistar rats were divided into four groups: OUA (1.8 µg/kg), saline (CTR), LPS (200 µg/kg), and OUA + LPS (OUA 20 min before LPS). The animals were sacrificed after 2 h, and the hippocampus was collected for analysis. After treatment, we determined the activities of Na,K-ATPase and glutamine synthetase (GS). In addition, expression of the α1, α2, and α3 isoforms of Na,K-ATPase and the glutamate transporters, EAAT1 and EAAT2, were also analysed. Treatment with OUA caused a specific increase in the α2 isoform expression (~20%), whereas LPS decreased its expression (~22%), and treatment with OUA before LPS prevented the effects of LPS. Moreover, LPS caused a decrease of approximately 50% in GS activity compared with that in the CTR group; however, OUA pre-treatment attenuated this effect of LPS. Notably, it was found that treatment with OUA caused an increase in the expression of EAAT1 (~30%) and EAAT2 (~25%), whereas LPS caused a decrease in the expression of EAAT1 (~23%) and EAAT2 (~25%) compared with that in the CTR group. When treated with OUA, the effects of LPS were abrogated. In conclusion, the OUA pre-treatment abolished the effect caused by LPS, suggesting that this finding may be related to the restoration of the interaction between FXYD2 and the studied membrane proteins.

Insecticide Resistance Status of Aedes albopictus (Diptera: Culicidae) Populations from Cuba.

Aedes albopictus (Skuse, 1894) is one of the major vectors for arboviruses such as dengue, Zika, and chikungunya. Originally from Southeast Asia, this species has spread to Africa, Europe, and the Americas, including Cuba. This spread has been favored by its great adaptability to variable temperatures and to the resistance of its eggs to desiccation. Chemical control of mosquitoes is an essential alternative to stop arbovirus transmission, but insecticide resistance status of the Cuban Ae. albopictus populations is unknown. For this study, Ae. albopictus larvae and adults were collected from two municipalities in Havana, Cuba in 2019. Adult bioassays for deltamethrin, cypermethrin, lambda-cyhalothrin, chlorpyrifos, propoxur, and bendiocarb susceptibility were conducted according to CDC methodology. Larval bioassays for temephos susceptibility were performed following WHO protocols. Resistance profiles for α and ß-esterases, glutathione S-transferase (GST), and multifunction oxidases (MFO) pathways were constructed and analyzed. Resistance to temephos and deltamethrin was detected in Mulgoba and Plaza field populations, but resistance to lambda-cyhalothrin was only found in the Plaza colony. Plaza colony exhibited a higher expression level to all four metabolic enzymes and α-esterases and GTS were over-expressed in Mulgoba. The development of insecticide resistance in Cuban Ae. albopictus populations makes it imperative that we develop integrated control strategies to minimize the development of resistance and provide effective vector control that prevents the onset of arbovirus epidemics.

Characterization of Insecticide Resistance in Aedes aegypti from the Zoological Garden of Havana, Cuba.

Chemical control of Aedes aegypti continues to be an indispensable alternative to preventing dengue, Zika, and chikungunya outbreaks. The Havana Zoological Garden requires constant vigilance because its special characteristics help in the spread of the causal agents of diseases transmitted by mosquitoes, which put the health of visitors at risk. The goals of this study were to determine the level of susceptibility and insecticide resistance mechanisms in the Ae. aegypti population. Temephos susceptibility in larvae was evaluated with bioassays using the World Health Organization's methodology, and susceptibility of adult mosquitoes was determined by the impregnated bottle bioassay, recommended by the Centers for Disease Control and Prevention. Resistance mechanisms were determined with biochemical assays. Mosquito larvae from the Havana Zoo were found resistant to temephos, which was associated with the activity of the enzymes α- and ß-esterases and mixed function oxidases but not glutathione-S-transferase. Adult mosquitoes were susceptible to pyrethroid (lambda-cyhalothrin, deltamethrin, and cypermethrin), organophosphate (chlorpyrifos), and carbamate (bendiocarb). Temephos resistance detected in the mosquito population from the Havana Zoo is an alert for the Vector Control Program, which must take measures to manage their resistance, relying on the surveillance carried out by Cuba's medical entomology laboratories.

Bacillus thuringiensis A21, un aislado no tóxico de elevada actividad larvicida residual contra Aedes aegypti / Bacillus thuringiensis A21, a non-toxic isolate of high residual larvicidal activity against Aedes aegypti

Introducción: Los productos derivados de Bacillus thuringiensis se han empleado con éxito en el control de insectos. Sin embargo, la poca residualidad promueve la búsqueda de alternativas. En Cuba diferentes estudios informan la evaluación y selección de aislamientos de B. thuringiensis con actividad larvicida contra Aedes aegypti: el aislado A21 se destaca por su elevada actividad larvicida y ausencia de betaexotoxina. Objetivo: Evaluar la toxicidad/patogenicidad aguda oral y actividad larvicida residual del aislado A21. Métodos: La evaluación de la toxicidad/patogenicidad aguda oral del aislado A21 se estableció en el modelo biológico Rattus norvegicus. Se realizaron observaciones clínicas diarias de los animales y se evaluó el peso corporal. Se estimó la eliminación y la infectividad de B. thuringiensis mediante análisis de muestras de heces, y de fluidos y órganos, respectivamente. Para determinar la actividad larvicida residual del aislado A21 frente a larvas de Ae. aegypti se realizaron bioensayos con diferentes volúmenes de recambios de agua semanal (total, parcial, no recambio) en los recipientes. La mortalidad obtenida se calculó a las 24-72 h. Resultados: Con el aislado A21 no se detectó toxicidad/patogenicidad aguda oral en Rattus norvegicus. En el estudio de residualidad, la mortalidad larvaria a las 72 h se mantuvo elevada (80-100 por ciento) hasta la semana 24. A partir de la semana 25 la mortalidad larvaria disminuyó (p < 0,05). Conclusiones: Se evidencia la baja toxicidad y la elevada actividad larvicida residual del aislado A21 contra Ae. aegypti y lo convierten en un candidato promisorio para el desarrollo de biolarvicidas. Estos productos biológicos podrían contribuir a mejorar las estrategias de control del vector existentes en Cuba(AU)

ABSTRACT Introduction: Bacillus thuringiensis-based products have been successfully used for insect control. However, their low residuality promotes the search for alternatives. In Cuba, different studies have informed about the evaluation and selection of B. thuringiensis isolates with larvicidal activity against Aedes aegypti: isolate A21 highlights for its high larvicidal activity and absence of beta-exotoxins. Objective: To evaluate the acute oral toxicity/pathogenicity and the residual larvicidal activity of isolate A21. Methods: The evaluation of the acute oral toxicity/pathogenicity of isolate A21 was established in the animal model Rattus norvegicus. Daily clinical observations of the animals were carried out, and their body weight was evaluated. The elimination and infectivity of B. thuringiensis were estimated by analyzing feces, and fluids and organs samples, respectively. To determine the residual larvicidal activity of isolate A21 to Ae. aegypti larvae, bioassays were conducted with different volumes of weekly water changes (total, partial, no change) in the containers. Mortality was calculated at 24h-72h. Results: No acute oral toxicity/pathogenicity was detected with isolate A21 in Rattus norvegicus. In the study of residuality, at 72h larval mortality remained high (80 percent-100%) until week 24, decreasing from week 25 (p < 0.05). Conclusions: It is evidenced the low toxicity and high residual larvicidal activity of isolate A21 against Ae. aegypti, which makes it a promising candidate for the development of biolarvicides. These biological products could contribute to the improvements of the existing vector control strategies in Cuba(AU)

Eugenia uniflora, Melaleuca armillaris, and Schinus molle essential oils to manage larvae of the filarial vector Culex quinquefasciatus (Diptera: Culicidae).

Populations of Culex quinquefasciatus Say, 1823 (Diptera: Culicidae) have shown resistance to insecticides of the carbamate and organophosphate classes. The objective of this study was to assess the susceptibility of C. quinquefasciatus larvae to essential oils from leaves of Eugenia uniflora L., Melaleuca armillaris (Sol. ex Gaertn.) Sm., and Schinus molle L and C. quinquefasciatus larvae's biochemical responses after their exposure to these leaves. The essential oils were chemically analyzed by GC and GC/MS. First, the lethal concentration for 50% (LC50) values was estimated using different concentrations of essential oils and probit analysis. The larvae were exposed for 1 h at the LC50 estimated for each essential oil. The susceptibility of the larvae to essential oils was evaluated using the following biochemical parameters: concentrations of total protein and reduced glutathione; levels of production of hydrogen peroxide and lipid peroxidation; and the activity of the enzyme acetylcholinesterase (AChE). The main chemical constituents in E. uniflora were E-ß-ocimene, curzerene, germacrene B, and germacrone; in M. armillaris were 1,8-cineole and terpinolene; and in S. molle were sabinene, myrcene, and sylvestrene. The essential oils had LC50 values between 31.52 and 60.08 mg/L, all of which were considered effective. All of them also promoted changes in biochemical parameters when compared to the control treatment. The essential oils of S. molle and E. uniflora inhibited the activity of the AChE enzyme, and the essential oil of M. armillaris increased it. All essential oils had larvicidal activity against C. quinquefasciatus, but the essential oil of E. uniflora was the most efficient. Thus, the findings of the present study suggest that the essential oil of E. uniflora can be considered promising for the development of botanical larvicides.

Isquemia aguda del canal anal / Acute Ischemia of the Anal Canal

Introducción: La isquemia anal aguda con gangrena es una entidad infrecuente, con escasa literatura publicada al respecto, cuyas cifras de mortalidad descritas se sitúan en un 20 por ciento - 40 por ciento de los casos. Debe considerarse en ancianos con enfermedad aterosclerótica que presenten síntomas gastrointestinales inferiores y shock hipotensivo. Puede ser causada por oclusión vascular aguda, enfermedad vascular grave o un estado de bajo flujo, aunque en algunos casos se presenta sin enfermedad vascular preexistente. Objetivo: Presentar la literatura existente acerca del manejo diagnóstico y terapéutico de la isquemia de canal anal en base a un caso clínico diagnosticado y tratado de urgencia en el hospital "Marina Baixa". Caso clínico: Se presenta un caso de isquemia espontánea de canal anal en paciente de 76 años de edad con morbilidad cardiovascular asociada. Debuta como cuadro séptico sin evidencia de causa desencadenante. Conclusiones: Las pruebas endoscópicas y radiológicas deben realizarse con urgencia y la valoración quirúrgica está siempre justificada en estos pacientes debido a la alta tasa de mortalidad descrita en el manejo conservador del cuadro. En casos graves, la reanimación preoperatoria y cirugía urgente para resecar el segmento gangrenoso es necesaria. Sin embargo, el tratamiento de la proctitis isquémica aguda es controvertido y depende, en parte, del estado basal del paciente y los hallazgos clínicos, siendo importante examinar otras posibles etiologías de proctitis isquémica y así determinar qué pacientes necesitan intervención quirúrgica temprana en comparación con una actitud más conservadora(AU)

Introduction: Acute ischemic gangrene of the anus is an infrequent entity, with little published literature, whose reported mortality figures are 20-40 percent of cases. It should be considered in elderlies with atherosclerotic disease who present lower gastrointestinal symptoms and hypotensive shock. It can be caused by acute vascular occlusion, severe vascular disease, or a low-flow state, although in some cases it presents without pre-existing vascular disease. Objective: To present the existing literature on diagnostic and therapeutic management of ischemia of the anal canal upon the base of a clinical case diagnosed and treated as an urgency at Marina Baixa hospital. Clinical case: The case is presented of a 76-year-old patient with spontaneous ischemia of the anal canal and associated cardiovascular morbidity. It debuts as a septic condition without evidence of a triggering cause. Conclusions: Endoscopic and radiological tests should be performed urgently. Surgical assessment is always justified in these patients, due to the high mortality rate described in the conservative management of the condition. In severe cases, preoperative resuscitation and urgent surgery to resect the gangrenous segment is necessary. However, managment of acute ischemic proctitis is controversial and depends, in part, on the patient's baseline status and clinical findings, while it is important to examine other possible etiologies of ischemic proctitis and thus determine which patients need early surgical intervention compared to a more conservative attitude(AU)

Aportes científicos del Instituto de Medicina Tropical "Pedro Kouri" a la vigilancia de Aedes aegypti (Díptera: Culicidae) en Cuba, 1982-2020

RESUMEN Entre las enfermedades de mayor incidencia en el mundo transmitidas por los culícidos o mosquitos se encuentran las arbovirosis como dengue, chikungunya, zika y fiebre amarilla, las cuales el humano las adquiere a través de la picadura de Aedes aegypti (Linnaeus, 1762) y Aedes albopictus (Skuse, 1894). Los programas de vigilancia entomológica de Ae. aegypti en el mundo tienen en común determinar cambios en la distribución geográfica del mosquito, obtener medidas relativas de sus poblaciones a lo largo del tiempo, evaluar la cobertura y el impacto de las intervenciones antivectoriales, así como monitorear la susceptibilidad y la resistencia de sus poblaciones a los principales insecticidas usados en el control vectorial. En este trabajo se resume los principales resultados de las investigaciones desarrolladas por el departamento de control de vectores del Instituto de Medicina Tropical ¨Pedro Kouri¨ (IPK) como Laboratorio Nacional de Referencia (LNR) para contribuir al fortalecimiento de la vigilancia entomológica de Ae. aegypti desde 1981 hasta el 2020. Con este artículo se destaca el trabajo realizado al cumplirse en el 2021, 40 años del inicio de la llamada campaña de erradicación de Ae. aegypti implementada a raíz de la epidemia de dengue hemorrágico (en la nueva clasificación denominada dengue severo) registrada en Cuba en 1981.

ABSTRACT Arbovirus infections such as dengue, chikungunya, zika and yellow fever are among the mosquito-borne diseases with the highest incidence worldwide. These conditions are transmitted by the bite of Aedes aegypti (Linnaeus, 1762) and Aedes albopictus (Skuse, 1894) mosquitoes. Common goals of entomological surveillance programs for Ae. aegypti in the world are to determine changes in the geographic distribution of the mosquito, obtain data concerning their populations throughout time, evaluate the coverage and impact of antivector interventions, and monitor the susceptibility and resistance of mosquito populations to the main insecticides used for vector control. The paper summarizes the main results of studies conducted from 1981 to 2020 by the Vector Control Department at Pedro Kourí Tropical Medicine Institute (IPK) as a National Reference Laboratory (NRL), to contribute to the strengthening of entomological surveillance of Ae. aegypti. Recognition is also made of the work done for forty years as part of the so-called Ae. aegypti eradication campaign implemented in response to the dengue hemorrhagic fever epidemic (severe dengue in the new classification) occurring in Cuba in 1981.

Effects of Dog-Assisted Therapy in Adolescents with Eating Disorders: A Study Protocol for a Pilot Controlled Trial.

BACKGROUND: Eating disorders are characterized by a persistent disturbance that alters food intake and it is often accompanied by anxiety, depression, low self-esteem, or reduced functional capacity and quality of life. Animal-assisted therapies (AAT) have shown benefits in these variables in children and adult populations. Thus, the present pilot study will aim to evaluate the effects of a dog-assisted therapy on the eating disorders symptoms, mental, psychosocial, and physical health, quality of life, and handgrip strength of adolescents suffering from eating disorders. METHODS: The current pilot study will involve 32 patients, who will be assigned to a control or an experimental group. Intervention will be conducted once a week for seven weeks. Neither the experimental nor the control group will discontinue their usual care. The main outcome measures will be the eating disorder symptoms and the health-related quality of life measured with standardized questionnaires, while the secondary variables will be anxiety, depression, character, behavior, strength, and body mass. CONCLUSIONS: This pilot-controlled trial will be the first to evaluate the effects of dog-assisted therapy on the physical and mental health of adolescents with eating disorders. Significant improvements, in the primary and secondary outcomes, may be expected based on the known benefits of AAT on self-esteem, stress, and self-control in different populations. Finally, although the program is focused on the improvement of adolescents' health, animal welfare will be a priority in this study.

Evaluation of Cardiotonic Steroid Modulation of Cellular Cholesterol and Phospholipid.

We have previously shown that 21-benzylidene digoxin (21-BD) increases the total cholesterol and phospholipid content on the membrane of HeLa cells. Lipid modulation caused by cardiotonic steroids (CTS) is still unexplored. Therefore, the aim of the present study was to evaluate the cholesterol and phospholipid modulation of the cell membrane caused by ouabain and 21-BD and the possible involvement of the caveolae on this modulation. For this, one cell line containing caveolae (HeLa) and other not containing (Caco-2) were used. The modulation of the lipid profile was evaluated by total cholesterol and phospholipids measurements, and identification of membrane phospholipids by HPTLC. The cholesterol distribution was evaluated by filipin staining. The caveolin-1 expression was evaluated by Western Blotting. Ouabain had no effect on the total membrane lipid content in both cell lines. However, 21-BD increased total membrane phospholipid content and had no effect on the membrane cholesterol content in Caco-2 cells. CTS were not able to alter the specific phospholipids content. In the filipin experiments, 21-BD provoked a remarkable redistribution of cholesterol to the perinuclear region of HeLa cells. In Caco-2 cells, it was observed only a slight increase in cholesterol, especially as intracellular vesicles. The caveolin-1 expression was not altered by any of the compounds. Our data mainly show different effects of two cardiotonic steroids. Ouabain had no effect on the lipid profile of cells, whereas 21-BD causes important changes in cholesterol and phospholipid content. Therefore, the modulation of cholesterol content in the plasma membrane of HeLa cells is not correlated with the expression of caveolin-1.

Cytotoxic effect of carbohydrate derivatives of digitoxigenin involves modulation of plasma membrane Ca2+ -ATPase.

Cardiac glycosides, such as digoxin and digitoxin, are compounds that interact with Na+ /K+ -ATPase to induce anti-neoplastic effects; however, these cardiac glycosides have narrow therapeutic index. Thus, semi-synthetic analogs of digitoxin with modifications in the sugar moiety has been shown to be an interesting approach to obtain more selective and more effective analogs than the parent natural product. Therefore, the aim of this study was to assess the cytotoxic potential of novel digitoxigenin derivatives, digitoxigenin-α-L-rhamno-pyranoside (1) and digitoxigenin-α-L-amiceto-pyranoside (2), in cervical carcinoma cells (HeLa) and human diploid lung fibroblasts (Wi-26-VA4). In addition, we studied the anticancer mechanisms of action of these compounds by comparing its cytotoxic effects with the potential to modulate the activity of three P-type ATPases; Na+ /K+ -ATPase, sarco/endoplasmic reticulum Ca2+ -ATPase (SERCA), and plasma membrane Ca2+ -ATPase (PMCA). Briefly, the results showed that compounds 1 and 2 were more cytotoxic and selectivity for HeLa tumor cells than the nontumor cells Wi-26-VA4. While the anticancer cytotoxicity in HeLa cells involves the modulation of Na+ /K+ -ATPase, PMCA and SERCA, the modulation of these P-type ATPases was completely absent in Wi-26-VA4 cells, which suggest the importance of their role in the cytotoxic effect of compounds 1 and 2 in HeLa cells. Furthermore, the compound 2 inhibited directly erythrocyte ghosts PMCA and both compounds were more cytotoxic than digitoxin in HeLa cells. These results provide a better understanding of the mode of action of the synthetic cardiac glycosides and highlights 1 and 2 as potential anticancer agents.

The γ-Benzylidene Digoxin Derivative BD-15 Increases the α3-Na, K-ATPase Activity in Rat Hippocampus and Prefrontal Cortex and no Change on Heart.

Our study aimed to investigate the effects of the new cardiotonic steroid BD-15 (γ-benzylidene derivatives) in the behavioral parameters, oxidative stress and the Na, K-ATPase activity in the hippocampus, prefrontal cortex and heart from rats to verify the safety and possible utilization in brain disorders. For this study, groups of male Wistar rats were used after intraperitoneal injection of 20, 100 and 200 µg/Kg with BD-15. The groups were treated for three consecutive days and the control group received 0.9% saline. BD-15 did not alter behavior of rats treated with different doses. An increase in the specific α2,3-Na, K-ATPase activity was observed for all doses of BD-15 tested in the hippocampus. However, in the prefrontal cortex, only the dose of 100 µg/Kg increased the activity of all Na, K-ATPase isoforms. BD-15 did not cause alteration in the lipid peroxidation levels in the hippocampus, but in the prefrontal cortex, a decrease of lipid peroxidation (~ 25%) was observed. In the hippocampus, GSH levels increased with all doses tested, while in the prefrontal cortex no changes were found. Subsequently, when the effect of BD-15 on cardiac tissue was analyzed, no changes were observed in the tested parameters. BD-15 at a dosage of 100 µg/Kg proved to be promising because it is considered therapeutic for brain disorders, since it increases the activity of the α3-Na, K-ATPase in the hippocampus and prefrontal cortex, as well as decreasing the oxidative stress in these brain regions. In addition, this drug did not cause changes in the tissues of the heart and kidneys, preferentially demonstrating specificity for the brain.

New bufadienolides extracted from Rhinella marina inhibit Na,K-ATPase and induce apoptosis by activating caspases 3 and 9 in human breast and ovarian cancer cells.

Bufadienolide compounds have been used for growth inhibition and apoptosis induction in tumor cells. Those families of cardiotonic steroids can bind the Na,K-ATPase, causing its inhibition. The use of bufadienolides is widely described in the literature as an anticancer function. The aim of this study was to evaluate the effects of bufadienolides and alkaloid isolated from venom samples from R. marina on tumor cells. We performed cytotoxicity assay in MDA-MB-231 and TOV-21G cells and evaluated the activity of Caspases (3 and 9), Na, K-ATPase, PMCA and SERCA. Four compounds were extrated from the venom of R. marina. The compound 1 showed higher cytotoxicity in MDA-MB-231cells. Compound 1 also showed activation of Caspase 3 and 9. This compound caused an inhibition of the activity and expression of Na, K-ATPase, and also showed activation of both caspase-9 and caspase-3 in MDA-MB-231 cells. We also observed that Compound 1 had a direct effect on some ATPases, such as Na, K-ATPase, PMCA and SERCA. Compound 1 was able to inhibit the activity of the purified Na, K-ATPase enzyme from the concentration of 5 µM. It also caused inhibition of PMCA at all concentrations tested (1 nM-30 µM). However, the compound 1 led to an increase of the activity of purified SERCA between the concentrations of 7.5-30 µM. Thus, we present a Na, K-ATPase and PMCA inhibitor, which may lead to the activation of caspases 3 and 9, causing the cells to enter into apoptosis. Our study suggests that compound 1 may be an interesting molecule as an anticancer agent.

Evaluation of the effect of cafeteria diet on the kidney Na,K-ATPase activity, and oxidative stress.

In this study, renal tissue, subdivided into the cortex and medulla of Wistar rats subjected to a cafeteria diet (CAF) for 24 days or to normal diet, was used to analyze whether the renal enzyme Na,K-ATPase activity was modified by CAF diet, as well as to analyze the α1 subunit of renal Na,K-ATPase expression levels. The lipid profile of the renal plasma membrane and oxidative stress were verified. In the Na,K-ATPase activity evaluation, no alteration was found, but a significant decrease of 30% in the cortex was detected in the α1 subunit expression of the enzyme. There was a 24% decrease in phospholipids in the cortex of rats submitted to CAF, a 17% increase in cholesterol levels in the cortex, and a 23% decrease in the medulla. Lipid peroxidation was significantly increased in the groups submitted to CAF, both in the cortical region, 29%, and in the medulla, 35%. Also, a reduction of 45% in the glutathione levels was observed in the cortex and medulla with CAF. CAF showed a nearly two-fold increase in glutathione peroxidase (GPX) activity in relation to the control group in the cortex and a 59% increase in the GPx activity in the medulla. In conclusion, although the diet was administered for a short period of time, important results were found, especially those related to the lipid profile and oxidative stress, which may directly affect renal function.

Evaluación de la eficacia larvicida de Rapidall NP3 (Bacillus thuringiensis) contra Aedes aegypti (Linnaeus) (Diptera: Culicidae) en condiciones de laboratorio / Evaluation of the larvicidal efficacy of Rapidall NP3 (Bacillus thuringiensis) against Aedes aegypti (Linnaeus) (Diptera: Culicidae) in laboratory conditions

Los mosquitos son vectores trasmisores de enfermedades como: dengue, zika y chikunguya. El control biológico es una alternativa viable a tener en cuenta por los programas de control. Bacillus thuringiensis es el microorganismo más usado en salud pública. Su efectividad como larvicida contra culícidos depende, en gran medida, de la eficiencia del proceso productivo. El objetivo es evaluar y comparar la eficacia larvicida contra Aedes aegypti de la formulación en polvo Rapidall NP3 con la del producto Bactivec®, formulación líquida utilizada en Cuba. Se realizaron bioensayos de laboratorio según los procedimientos descritos y sugeridos por la OMS para determinar la eficacia, exponiendo las larvas a las concentraciones recomendadas por los fabricantes de los productos Rapidall NP3 y Bactivec®. A los recipientes utilizados se les realizó recambio de agua medio, total y sin recambio, con una frecuencia semanal. A las 24 h se calculó la mortalidad obtenida y se analizó estadísticamente mediante un ANOVA de una vía; cuando existieron diferencias significativas se aplicó un análisis Post-hoc mediante la prueba de Tukey. El producto evaluado Rapidall NP3, provocó 100 por ciento de mortalidad larvaria durante 11 semanas, independientemente de la proporción de recambio de agua, mientras que el Bactivec® solo provocó 100 por ciento de mortalidad hasta las 6 semanas de iniciado el experimento. El biolarvicida Rapidall NP3 mostró buena eficacia y actividad residual prolongada al compararlo con Bactivec®, por lo que recomendamos su evaluación en el terreno en el control larvario de depósitos que constituyen sitios de cría habituales y que no pueden ser eliminados físicamente(AU)

Mosquitoes are vectors of human diseases such as dengue, zika and chikungunya. Biological control is a viable alternative to be taken into account in control programs. Bacillus thuringiensis is the microorganism most commonly used in public health. Its effectiveness as a larvicide against culicides depends to a great extent to the production process. The objectives of the study were to evaluate and compare the larvicidal efficacy against Aedes aegypti of the Rapidall NP3 powder formulation with Bactivec®, a liquid formulation used in Cuba. Laboratory bioassays were conducted to determine efficacy following WHO protocols. Larvae were exposed to the concentrations recommended by Rapidall NP3 and Bactivec® manufacturers. The water in the containers used in the study was replaced once a week: half, whole and no replacement. At 24 hours mortality was estimated and statistically analysis were done by one-way ANOVA. When significant differences were found, post-hoc analysis was performed with Tukey's test. Rapidall NP3 caused 100 percent larval mortality for 11 weeks, regardless of the water replacement proportion, whereas Bactivec® caused 100 percent mortality for only 6 weeks after the start of the experiment. When compared with Bactivec®, biolarvicide Rapidall NP3 displayed better efficacy and more prolonged residual activity. We therefore recommend its field evaluation for larval control of containers constituting habitual breeding sites which cannot be physically eliminated(AU)

Ouabain attenuates oxidative stress and modulates lipid composition in hippocampus of rats in lipopolysaccharide-induced hypocampal neuroinflammation in rats.

Our study aimed to analyze the effect of ouabain (OUA) administration on lipopolysaccharide (LPS)-induced changes in hippocampus of rats. Oxidative parameters were analyzed in Wistar rats after intraperitoneal injection of OUA (1.8 µg/kg), LPS (200 µg/kg), or OUA plus LPS or saline. To reach our goal, activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX), in addition to levels of reduced glutathione (GSH), protein carbonyl (PCO) and lipid peroxidation (LPO) were evaluated. We also analyzed the membrane lipid profile and some important lipids for the nervous system, such as phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylinositol (PI), phosphatidic acid and sphingomyelin. The group that received only LPS showed increased oxidative stress, as evidenced by an increase in LPO (about twice), PCO (about three times) levels, and CAT activity (80%). Conversely, administration of LPS decreased GSH levels (55%), and GPx activity (30%), besides a reduction in the amount of PI (60%) and PC (45%). By other side, OUA alone increased the amount of PI (45%), PE (85%), and PC (70%). All harmful effects recorded were attenuated by OUA, suggesting a protective effect against LPS-induced oxidative stress. The relevance of our results extends beyond changes in oxidative parameters induced by LPS, because nanomolar doses of OUA may be useful in neurodegenerative models. Other studies on other cardenolides and substances related issues, as well as the development of new molecules derived from OUA, could also be useful in general oxidative and/or cellular stress, a condition favoring the appearance of neuronal pathologies.

Síndrome de embriopatía diabética: reporte de caso / Diabetic embriopaty. Case report

Antecedentes. Las malformaciones congénitas producidas por la diabetes materna constituyen un síndrome conocido como «embriopatía diabética¼. Los defectos se asocian a diabetes pregestacional, tipo 1 ó 2. Su patogénesis no está totalmente esclarecida pero la hiperglicemia parece ser el estímulo intrauterino más importante, favoreciendo producción de radicales libres y otras condiciones adversas para la implantación y desarrollo embrionario/fetal. Descripción del caso. Presentamos caso de gestante multípara, procedente de Tegucigalpa, M.D.C, antecedente de diabetes mellitus tipo 2, hipotiroidismo y obesidad grado I, historia gineco-obstétrica: G:3, C:2, A:1, HV:1, HM:1, 6 controles prenatales, con embarazo de 30.6 SG X FUM. Acude a control prenatal, realizándose ultrasonido obstétrico que mostró raquisquisis con hidrocefalia severa y tendencia al macrocráneo, pie equinovaro bilateral y polihidramnios (ILA 21.7cc). Paciente refirió aproximadamente 10 contracciones esporádicas en 24 hrs. Examen físico: GO: cérvix dilatado 2 cm, B: 50% AP: -3, membranas íntegras, cefálico. Es ingresada con plan de cesárea más OTB obteniendo RN con múltiples malformaciones congénitas (defectos del tubo neural, estructuras craneofaciales, y óseas). Se ingresó a unidad de cuidados intermedios, realizándose USG transfontanelar que reportó hidrocefalia severa y por la presencia de mielomeningocele, se sospechóArnold Chiari tipo II. Se realizó mieloplastía con técnica habitual sin complicaciones, paciente permaneció estable hasta el momento de su egreso hospitalario. Conclusión/recomendación. Niveles de HbA1c en el primer trimestre ≥7% o Glucosa basal >120mg/dl se han asociado al desarrollo de malformaciones. Es importante un estricto control glicémico preconcepcional para disminuir la incidencia de malformaciones asociadas a hiperglicemia en el embarazo...(AU)

Susceptibilidad de diferentes estadios larvarios de Aedes albopictus (S) (Diptera: Culicidae) a la infección por dos especies de nematodos mermítidos en condiciones de laboratorio / Sensitivity of different larval stages of Aedes albopictus (S) (Diptera: Culicidae) to infection by two species of mermithids nematodes in laboratory conditions

Introducción: La combinación de insecticidas, la reducción de criaderos y el control biológico se proponen como una alternativa para el control de culícidos. Los nematodos entomoparásitos, Romanomermis iyengari y Strelkovimermis spiculatus, han sido empleados con éxito en estudios de laboratorio y campo, pero aún no se han evaluado en Cuba contra larvas de Aedes albopictus, mosquito vector de dengue, chikungunya, zika y otras arbovirosis. Objetivo: Determinar la susceptibilidad de tres estadios larvales deAe. albopictus a la infección de R. iyengari y S. spiculatus en condiciones de laboratorio. Métodos: Se aplicó una dosis de 7:1 (preparásitos/larva) a 100 larvas de Ae. albopictus en diferentes estadios de desarrollo (I, II y III). Después de 96 h de la posinfección, se tomaron al azar 30 larvas y se disectaron para determinar la intensidad y frecuencia de la infección. Cada estadio larval de Ae. albopictus por especie de nematodo, constó de 12 réplicas y se colocaron controles. Resultados: S. spiculatus mostró mayor frecuencia de la infección en los estadios larvales estudiados (97,22 por ciento; 96,66 por ciento y 73,33 por ciento) que R. iyengari (90,27 por ciento; 83,61 por ciento y 57,22 por ciento), mientras que la intensidad de la infección por estadios fue de 2,54; 2,44 y 1,55 para S. spiculatus y 2,39; 2,46 y 1,97 para R. iyengari. Conclusiones: Aunque S. spiculatus fue superior a R. iyengari, con mayores valores de frecuencia de infección en los tres estadios, ambas especies demostraron ser buenos candidatos como agentes de biocontrol de Ae. Albopictus; se recomiendan estudios posteriores antes de su uso en condiciones naturales(AU)

Introduction: The combination of safe insecticides for the environment, breeding sites´ reduction and biological control is proposed as a promising approach for Culicids control. The enthomoparasites nematodes Romanomermis iyengari and Strelkovimermis spiculatus have been used successfully in laboratory and field studies, but haven't been evaluated yet in Cuba against Aedes albopictus larvae. This mosquito species constitutes a vector of dengue, chikungunya, zika and other arboviruses. Objective: To determine the sensitivity of three larval stages ofAe. albopictus to the infection of R. iyengari and S. spiculatus in laboratory conditions. Methods: A dose of 7: 1 (preparasitics / larva) was applied to 100 larvae of Ae. albopictus in different stages of development (I, II and III). 96 hours after the post-infection, 30 larvae were taken at random and dissected to determine the intensity and frequency of the infection. Each Ae. Albopictus larval stage had twelve replicas by stages and controls were made to them. Results: S. spiculatus showed higher frequency of infection in the larval stages studied (97.22 percent, 96.66 percent and 73.33 percent) than R. iyengari (90.27 percent, 83.61 percent and 57.22 percent), while the intensity of infection by stages was 2.54; 2.44 and 1.55 for S.spiculatus and 2.39; 2.46 and 1.97 for R.iyengari. Conclusions: Although S. spiculatus was superior to R. iyengari, with higher values of infection frequency in the three stages, both species proved to be good candidates as bio-control agents of Ae. albopictus and subsequent studies are recommended before use in natural conditions(AU)