Use of coumarins as complementary medicine with an integrative approach against cervical cancer: background and mechanisms of action.

Cervical cancer is characterized by the cellular transformation caused by Human Papillomavirus (HPV), favoring cell proliferation, migration, invasion, and metastasis. Cervical cancer is conventionally treated with radiation therapy, and chemotherapy focused on the destruction of tumor cells. However, chemoresistance and low selectivity between tumor and non-tumor cells have been reported, causing side effects in patients. Metabolites of natural origin have shown selectivity against tumor cells, suggesting their use for reducing the side effects caused by drugs used in conventional therapy. Among these compounds, several natural coumarins stand out, such as auraptene, scopoletin, osthole, and praeruptorin, of which antiproliferative, anti-migratory, and anti-invasive activity have been reported. Auraptene, scopoletin, osthole, and praeruptorin show a cytotoxic or antiproliferative effect on cervical tumor cells, arresting the cell cycle by inducing the overexpression of negative regulators of the cell cycle, or inducing cell death by increasing the expression of pro-apoptotic proteins and decreasing that of anti-apoptotic proteins. On the other hand, auraptene, scopoletin, and praeruptorin inhibit the capacity for migration, invasion, and metastasis of cervical tumor cells, mainly by inhibiting the expression and activity of matrix metalloproteinase-2 and -9. The PI3K/Akt signal pathway appears to be central to the anti-tumor activity of the coumarins analyzed in this review. In addition, auraptene, osthole, and praeruptorin are useful in sensitizing tumor cells to radiotherapy or chemotherapeutic molecules, such as FOLFOX, cisplatin, or DOX. Coumarins offer an excellent possibility for developing new drugs as complementary medicine with an integrative approach against cervical cancer.

Estudio clínico y micológico de otomicosis en diabéticos: una serie de 17 casos / Clinical and mycological study of otomycosis in diabetics: series of 17 cases

RESUMEN Introducción: La otomicosis en pacientes inmunosuprimidos esta caracterizada por ser bilateral y ser causada por candida. Pocos estudios comparan las características micológicas encontradas en la microscopía directa y el cultivo. Objetivo: Identificar las características clínicas y micológicas de la otomicosis en pacientes diabéticos. Material y método: Estudio transversal en centro hospitalario de segundo nivel. Criterios de inclusión: pacientes diabéticos con diagnóstico clínico de otomicosis. Intervención: la muestra se examinó directamente bajo el microscopio y se cultivó. Resultados: Se incluyeron 17 pacientes, 10 mujeres y 7 hombres con una edad media de 47,5 años. Los síntomas predominantes fueron hipoacusia en 91,4% (n =16), prurito en 82,4% (n =14), otorrea en 76,5% (n=13)y otalgia en 70,6% (n =12). Afección bilateral se encontró en 47,1% (n =8). Estudio directo al microscopio mostró levaduras en 94,1% (n =16) y 5,9% mostró aspergillus (n =1). Cándida fue el género más comúnmente encontrado en los cultivos y en el examen directo microscópico con 94,1% (n =16) y Candida albicans la especie más común con 88,2% (n =15). Conclusión: Candida albicans es el agente etiológico más común en pacientes diabéticos con otomicosis. Su presentación clínica más frecuente es hipoacusia, prurito y otorrea. El examen directo identificó adecuadamente a los géneros fúngicos.

ABSTRACT Introduction: Otomycosis in immunocompromised patients is characterize by its bilateral course and the predominant etiologic agent is Candida. Few studies compare the mycological features between microscopic direct exam and culture. Aim: To identify the clinical and mycological characteristics of otomycosis in diabetic patients. Material and method: Transversal study. Secondary care center. Inclusion criteria: diabetic patients with clinical diagnosis of otomycosis. Intervention: Direct examination under a microscope of the ear sample and culture. Results: We included 17 patients, 10 women, 7 men with a mean age of 47.5 years. Symptoms were hearing loss 94.1% (n = 16), pruritus 82.4% (n =14) otorrhoea 76.5% (n =13) and otalgia 70.6% (n =12). Bilateral involvement was found in 47.1% (n =8). Direct microscopic study found 94.1% of yeast (n =16) and 5.9% of Aspergillus (n =1). Candida was the most common fungal genus in culture and microscopic exam with 94.1% (n =16) of cases and Candida albicans was the most common species in 88.2% (n =15) cases. Conclusion: Candida albicans is the most common etiologic agent in diabetic patients with otomycosis. Main symptoms were hearing loss, itching and otorrhea. Direct exam correctly identified the fungal genus.

Fatty acid profile of goat milk in diets supplemented with chia seed (Salvia hispanica L.).

Chia seed (Salvia hispanica L.) is the greatest known plant source of n-3 α-linolenic acid. The present study evaluated the effects of 3 inclusion levels of chia seed [zero (control); low, 2.7% (CLow); and high, 5.5% (CHigh)] in diets of dairy goats on milk yield and fatty acid profile. Nine Saanen dairy goats in the last third of lactation period, live weight 38 ± 8.7 kg, housed in metabolic cages, were fed iso-proteic and iso-energetic (160 g of crude protein/d and 11 MJ of metabolizable energy/d) diets. Gas chromatography was used to analyze fatty acid profile and total conjugated linoleic acid (CLA). Silver ion HPLC was used to analyze the isomeric profile of CLA. The results were subjected to variance analysis using a Latin square design repeated 3 × 3. The CHigh treatment was higher for dry matter, neutral detergent fiber, and acid detergent fiber intake compared with CLow and control diets. Digestibility was not affected by the inclusion of chia seeds. The CHigh diet improved N intake with respect to the control and CLow diet. Milk yield and chemical composition were not affected by the treatment. The milk fatty acid profile of C18:0, C18:1, C18:2, and C:20 was higher for CHigh than the other treatments. The in vitro gas production (mL of gas/g of dry matter) was lower in CHigh than the control diet. In conclusion, the addition of chia seeds at the CHigh level in dairy goat diets negatively affected in vitro rumen fermentation, but increased the milk fatty acid profile of C18:0, C18:1n-9 cis, and C:20, monounsaturated fatty acids, and polyunsaturated fatty acids. The total CLA content increased from 0.33 to 0.73% with the supplementation of chia to the diet, as well as the isomers cis-9,trans-11, trans-7,cis-9, trans-11,cis-13, and trans-12,trans-14.

Apoptotic activities of cardenolide glycosides from Asclepias subulata.

ETHNOPHARMACOLOGICAL RELEVANCE: Asclepias subulata Decne. (Apocynaceae) is a shrub occurring in Sonora-Arizona desert. The ethnic groups of Sonora, Mexico, Seris and Pimas, use this plant for the treatment of sore eyes, gastrointestinal disorders and cancer. AIM OF THE STUDY: To determine the cell death pathways that the cardenolide glycosides with antiproliferative activity found in the methanol extract of A. subulata are able to activate. MATERIALS AND METHODS: The effect of cardenolide glycosides isolated of A. subulata on induction of apoptosis in cancer cells was evaluated through the measuring of several key events of apoptosis. A549 cells were treated for 12h with doses of 3.0, 0.2, 3.0 and 1.0µM of 12, 16-dihydroxicalotropin, calotropin, corotoxigenin 3-O-glucopyranoside and desglucouzarin, respectively. Apoptotic and necrotic cell levels were measured by double staining with annexin V-FITC/PI. Mitochondrial membrane depolarization was examined through JC-1 staining. Apoptosis cell death and the apoptosis pathways activated by cardenolide glycosides isolated of A. subulata were further characterized by the measurement of caspase-3, caspase-8 and caspase-9 activity. RESULTS: Apoptotic assays showed that the four cardenolide glycosides isolated of A. subulata induced apoptosis in A549 cells, which was evidencing by phosphatidylserine externalization in 18.2%, 17.0%, 23.9% and 22.0% for 12, 16-dihydroxicalotropin, calotropin, corotoxigenin 3-O-glucopyranoside and desglucouzarin, respectively, compared with 4.6% of control cells. Cell death was also associated with a decrease in mitochondrial membrane potential, which was more than 75% in the treated cultures respect to control. The activation of caspase-3 was observed in all cardenolide glycosides-treated cancer cells indicating the caspase-dependent apoptosis of A549 cells. Extrinsic and intrinsic apoptosis pathways were activated by cardenolide glycosides treatment at the doses tested. CONCLUSION: In this study was found that cardenolide glycosides, 12, 16-dihydroxicalotropin, calotropin, corotoxigenin 3-O-glucopyranoside and desglucouzarin, isolated from A. subulata induced the cell death trough caspase-dependent apoptosis, which was activated, preferably, by extrinsic pathway.

Antiproliferative activity of cardenolide glycosides from Asclepias subulata.

ETHNOPHARMACOLOGICAL RELEVANCE: Asclepias subulata Decne. is a shrub occurring in Sonora-Arizona desert (Mexico-USA). The ethnic groups, Seris and Pimas, use this plant for the treatment of sore eyes, gastrointestinal disorders and cancer. AIM OF THE STUDY: To isolate the compounds responsible for antiproliferative activity of the methanol extract of A. subulata. MATERIALS AND METHODS: A bioguided fractionation of methanol extract of A. subulata was performed using MTT assay to measure the antiproliferative activity of different compounds on three human cancer cell lines (A549, LS 180 and PC-3), one murine cancer cell line (RAW 264.7) and one human normal cell line (ARPE-19). The methanol extract was partitioned with hexane, ethyl acetate and ethanol. The active fractions, ethanol and residual, were fractioned by silica-column chromatography and active sub-fractions were separated using HPLC. The chemical structures of isolated compounds were elucidated with different chemical and spectroscopic methods. RESULTS: A new cardenolide glycoside, 12, 16-dihydroxycalotropin, and three known, calotropin, corotoxigenin 3-O-glucopyranoside and desglucouzarin, were isolated of active sub-fractions. All isolated compounds showed a strong antiproliferative activity in human cancer cells. Calotropin was the more active with IC50 values of 0.0013, 0.06 and 0.41 µM on A549, LS 180 and PC-3 cell lines, respectively; while 12, 16-dihydroxycalotropin reached values of 2.48, 5.62 and 11.70 µM, on the same cells; corotoxigenin 3-O-glucopyranoside had IC50 of 2.64, 3.15 and 6.62 µM and desglucouzarin showed values of 0.90, 6.57 and 6.62, µM. Doxorubicin, positive control, showed IC50 values of 1.78, 6.99 and 3.18 µM, respectively. The isolated compounds had a weak effect on murine cancer cells and human normal cells, exhibiting selectivity to human cancer cells. CONCLUSION: In this study, we found that 12, 16-dihydroxicalotropin, calotropin, corotoxigenin 3-O-glucopyranoside and desglucouzarin are responsible of antiproliferative properties of A. subulata, and that these compounds are highly selective to human cancer cells. Further studies are needed in order to establish the action mechanisms of the isolated compounds.

Aggregatibacter actinomycetemcomitans QseBC is activated by catecholamines and iron and regulates genes encoding proteins associated with anaerobic respiration and metabolism.

Aggregatibacter actinomycetemcomitans QseBC regulates its own expression and is essential for biofilm growth and virulence. However, the signal that activates the QseC sensor has not been identified and the qseBC regulon has not been defined. In this study, we show that QseC is activated by catecholamine hormones and iron but not by either component alone. Activation of QseC requires an EYRDD motif in the periplasmic domain of the sensor and site-specific mutations in EYRDD or the deletion of the periplasmic domain inhibits catecholamine/iron-dependent induction of the ygiW-qseBC operon. Catecholamine/iron-dependent induction of transcription also requires interaction of the QseB response regulator with its binding site in the ygiW-qseBC promoter. Whole genome microarrays were used to compare gene expression profiles of A. actinomycetemcomitans grown in a chemically defined medium with and without catecholamine and iron supplementation. Approximately 11.5% of the A. actinomycetemcomitans genome was differentially expressed by at least two-fold upon exposure to catecholamines and iron. The expression of ferritin was strongly induced, suggesting that intracellular iron storage capacity is increased upon QseBC activation. Consistent with this, genes encoding iron binding and transport proteins were down-regulated by QseBC. Strikingly, 57% of the QseBC up-regulated genes (56/99) encode proteins associated with anaerobic metabolism and respiration. Most of these up-regulated genes were recently reported to be induced during in vivo growth of A. actinomycetemcomitans. These results suggest that detection of catecholamines and iron by QseBC may alter the cellular metabolism of A. actinomycetemcomitans for increased fitness and growth in an anaerobic host environment.

Hipodermoclisis como alternativa para la rehidratación en niños / Hypodermoclysis as an alternative to pediatric rehydration / Hipodermóclise como alternativa para a reidratação em crianças

La deshidratación es una de las principales causas de mortalidad en la población pediátrica en países en vía de desarrollo. Por ello, se han estudiado alternativas de manejo para la rehidratación oral, intravenosa e intraósea, de fácil aplicación y bajo costo. La hipodermoclisis es la técnica para la administración subcutánea de líquidos con el fin de prevenir o tratar la deshidratación leve o moderada; olvidada por muchos años debido a la falta de experiencia y protocolos bien estructurados. Recientemente resurge el interés en investigar e implementar esta técnica gracias a sus beneficios y menores riesgos frente a las otras opciones, entre las que se encuentran facilidad de aplicación, mantenimiento de la vía, comodidad del paciente y disminución de complicaciones por daño endotelial, demostrados en diferentes estudios como es el caso del Infuse Peds 1 y 2. Para favorecer la absorción subcutánea se puede implementar la hialuronidasa recombinante humana, enzima encargada de disolver el ácido hialurónico para alterar, temporalmente, la permeabilidad del tejido, que es el objeto de análisis dentro de estos estudios. En Colombia esta técnica aún no ha sido completamente difundida para aplicarla en la población pediátrica. Es imperativo divulgar opciones de manejo que sean efectivas y de bajo costo para la atención de las enfermedades más prevalentes. La hipodermoclisis cumple con estos objetivos y cuenta, además, con beneficios como la satisfacción de los padres y mejora índices de morbimortalidad infantil.

Dehydration is one of the main causes of mortality in pediatric population in developing countries. Thus, we have studied alternative measures that are low-cost and easy to administer for oral, intravenous, and intraosseuos rehydration. Hypodermoclysis is the technique for subcutaneous administration of liquids in order to prevent or treat mild or moderate dehydration. This method has been forgotten for many years due to the lack of experience and well-structured protocols. Recently, there has been increasing interest in researching and implementing this technique due to the benefits and lower risks compared to other options, including easy administration, pathway maintenance, patient comfort, and reduced complications due to endothelial damage, demonstrated in different studies, such as INFUSE-Peds I and II. Human recombinant hyaluronidase can be implemented in order to favor subcutaneous absorption; this enzyme is in charge of dissolving hyaluronic acid to temporarily alter the permeability of the tissue, which is the object of analysis of these studies. This technique has not been completely disseminated in Colombia to administer in pediatric population. It is imperative to divulge treatment options that are effective and low-cost to treat the most prevalent diseases. Hypodermoclysis meets these objectives and also has benefits such as parent satisfaction and improves indexes of infant morbidity and mortality.

A desidratação é uma das principais causas de mortalidade na população pediátrica em países em via de desenvolvimento. Por isso, se estudaram alternativas de manejo para a reidratação oral, intravenosa e intraóssea, de fácil aplicação e baixo custo. A hipodermóclise é a técnica para a administração subcutânea de líquidos com o fim de prevenir ou tratar a desidratação leve ou moderada; esquecida por muitos anos devido à falta de experiência e protocolos bem estruturados. Recentemente ressurge o interesse em investigar e implementar esta técnica graças a seus benefícios e menores riscos frente às outras opções, entre as que se encontram facilidade de aplicação, manutenção da via, comodidade do paciente e diminuição de complicações por dano endotelial, demostrados em diferentes estudos como é o caso do Infuse Peds 1 e 2. Para favorecer a absorção subcutânea se pode implementar a hialuronidase recombinante humana, enzima encargada de dissolver o ácido hialurônico para alterar, temporalmente, a permeabilidade do tecido, que é o objeto de análise dentro destes estudos. Na Colômbia esta técnica ainda não há sido completamente difundida para aplicá-la na população pediátrica. É imperativo divulgar opções de manejo que sejam efetivas e de baixo custo para a atenção das doenças mais prevalentes. A hipodermóclise cumpre com estes objetivos e conta, ademais, com benefícios como a satisfação dos pais e melhora índices de morbimortalidade infantil.

Antibacterial and antifungal activities of some Mexican medicinal plants.

In Mexico about 4,000 plant species have some medicinal use. The aim of this work was to evaluate the antimicrobial activity of six Mexican medicinal plants against fungi and Gram-positive and Gram-negative bacteria. Methanolic extracts were prepared from the Mexican medicinal plants Amphypteringium adstrigens, Castella tortuosa, Coutarea latiflora, Ibervillea sonorae, Jatropha cuneata, and Selaginella lepidophylla. The antibacterial and antifungal activities of the plants were determined by the broth microdilution method and the radial growth inhibition assay, respectively. All Mexican plants tested showed antimicrobial activity. Among the six plant extracts analyzed, J. cuneata showed the highest growth-inhibitory activity against fungi, Gram-positive and Gram-negative bacteria (J. cuneata > A. adstrigens > C. latiflora > C. tortuosa > I. sonorae approximately S. lepidophylla). Shigella flexneri and Staphylococcus aureus were the most susceptible bacteria to plant extracts. Complete inhibition of S. flexneri growth was observed with J. cuneata methanolic extract at 90 microg/mL. This plant extract also showed the strongest antifungal activity against Fusarium verticillioides and Aspergillus niger. Our data suggest that the medicinal plants tested have important antimicrobial properties. This is the first report describing the antimicrobial activities of several of the Mexican medicinal plants used in this study.

La determinación peroperatoria de PTHi como factor predictivo de curación en el hiperparatiroidismo / Intraoperative intact parathyroid hormone determination as a predictive factor of hyperparathyroidism resolution

Introducción. Nuestros objetivos son valorar la eficacia de la determinación peroperatoria de la hormona paratiroidea intacta (PTHi) como marcador de curación del hiperparatiroidismo (HPT), establecer el número mínimo de muestras sanguíneas necesarias y comprobar si la manipulación cervical eleva la PTHi basal. Pacientes y método. Estudio prospectivo realizado con 45 pacientes. Peroperatoriamente se hicieron 3 determinaciones sanguíneas de PTHi: basal y a los 10 y 25 min de extirpar la lesión. Para analizar los efectos de la manipulación cervical, en 19 pacientes, se hicieron otras 2 determinaciones tras 2 min de masaje en ambos lados del cuello. Se aceptó como criterio diagnóstico y terapéutico el descenso > 50% del valor de PTHi entre las muestras postexeréticas y la basal (gradiente > 50) y como criterio de curación la normalización de la calcemia. Resultados. Siempre que se extirpó la causa del HPT hubo descensos de PTHi a los 10 y 25 min de la exéresis. Este descenso tuvo valor predictivo de curación cuando el gradiente fue > 50. La manipulación cervical (masaje) no produjo elevación de la cifra de PTHi. En los casos curados, también se normalizaron las cifras de la calcemia. Conclusiones. La determinación intraoperatoria de PTHi, con gradiente > 50, es un excelente indicador de curación. Sólo se precisan 2 muestras de PTHi: la basal y la extraída a los 10 min de la exéresis. La manipulación cervical preoperatoria no produjo elevación de los valores de PTHi (AU)

Introduction. The objectives of this study were: a) to evaluate the effectiveness of intraoperative intact parathyroid hormone (PTHi) determination as a marker of hyperparathyroidism resolution; b) to establish the minimum number of blood samples required; and c) to determine whether cervical manipulation increases baseline PTHi levels. Patients and method. We performed a prospective study in 45 patients. Three intraoperative blood PTHi determinations were performed: at baseline and at 10 and 25 minutes after excising the lesion. To analyze the effects of cervical manipulation, in 19 patients, 2 further determinations were made after 2 minutes of massage on both sides of the neck. A decrease of > 50% in PTHi values between postexeresis samples and the baseline sample (gradient > 50%) was used as diagnostic and therapeutic criteria and normalization of calcemia was used as a criteria for complete resolution. Results. Whenever the lesion causing hyperparathyroidism was extirpated, PTHi levels decreased at 10 and 25 minutes after exeresis. This decrease was predictive of complete resolution when the gradient was > 50. Cervical manipulation (massage) did not increase PTHi values. In patients with complete resolution, blood calcium levels also returned to normal. Conclusions. 1. Intraoperative PTHi determination with a gradient > 50 is an excellent prognostic marker of resolution. 2. Only 2 PTHi samples are required: one at baseline and another at 10 minutes after exeresis. 3. Preoperative cervical manipulation does not increase PTHi values (AU)

Blood metabolites and their relationship with production variables in dual-purpose cows in Venezuela.

A survey was carried out on 79 lactating Bos taurus/indicus cross-bred cows on three dual-purpose cattle farms to measure the blood concentration of metabolites and to evaluate possible relationships with nutritional status and productive variables. A rotational grazing system on Star grass and other tropical pastures (10-12% CP in leaves) was used and 2-3 kg/cow/day of concentrate were fed on two farms. Restricted calf suckling was used in two herds. Average milk yield sold per farm was 6 kg/day/cow and body condition scores (BCS) were between 3.0 and 3.8 on a scale of one-to-five. On two farms, the average interval from calving to conception (ICC) was more than 145 days. Mean blood concentrations of albumin, globulin, urea, beta-hydroxybutyrate and phosphorus were generally within reference values, but a significant group of cows had low levels of albumin and urea and high levels of globulin. Packed cell volume (PCV) was below normal values, with anemia in 63% of cows during the second trimester of lactation, which was negatively correlated to milk yield. The high incidence of anemia could be related to factors such as hematophagic parasites, not evaluated in this study. ICC values were negatively related to albumin level and could be associated with protein deficiency in the diet or with disease, as globulin values were high in many cows. Based on these diagnoses, an experiment was carried out on one of the farms to evaluate the influence of supplementation with 0.5 kg/cow/day of fish meal. Total milk yield was not influenced by the fish meal and reproductive efficiency was high in the two supplemental treatments. It was shown that supplementation with undergraded protein is not required in these cows.

Identification of Entamoeba histolytica antigens recognized by IgA class human antibodies in sera and colostra of puerperal women using immunoblotting techniques.

Immunodetection of total antigens of Entamoeba histolytica trophozoites transferred to nitrocellulose paper was realized with IgA antibodies present in colostra and sera samples of 28 puerperal women and in 24 sera samples of patients with a diagnosis of amebic hepatic abscess. The most frequently recognized antigens by colostra antibodies were those which correspond to 1290, 82.79, 52.14, 40.91, 37.75, 35.08, 33.52, 28.53, 27.53 and 25.32 KDa of molecular weight, in the case of puerperal sera those of 82.79, 52.14, 40.91, 37.75, 35.08, 33.52, 28.53, 27.53, and 25.32 KDa, whereas the sera of patients with amebic hepatic abscess those of molecular weight of 719, 118.45, 88.45, 69.45, 55.26, 52.14, 49.38, 42.72, 36.81, 35.08, 33.52, 25.32, 23.82, 23.47, 20.52 and 20.03 KDa. The putative adhesins were recognized infrequently by Iga of puerperal colostra and sera while the IgA of sera of patients with amebic hepatic abscess recognized those of 118.45 KDa (112 KDa in other studies) in 83.33% of the cases.

Wernicke's encephalopathy. A case report with neurophysiologic and CT-scan studies.

A case of Wernicke's encephalopathy with ataxia, confusion, memory loss, partial seizures of complex behavior and hypothermia, subsequent to thiamine depletion due to chronic malnourishment and triggered by an episode of acute vomiting and diarrhea, is reported, Computerized tomography (CT-scan) depicted small bilateral lesions in areas adjacent to the walls of the third ventricle, common location of the lesions seen in autopsy material of Wernicke's encephalopathy. Early diagnosis and treatment with vitamin B complex supplemented with intensive mnemonic and cognitive therapy led to complete recovery in a ten day period.