T cell receptor excision circles as a tool for evaluating thymic function in young children

The thymus is a primary lymphoid organ responsible for the maturation of T cells as well as the immunological central tolerance. It is in the antenatal period and infancy that it plays its major role. In clinical practice, T cell receptor excision circles (TRECs) are considered a direct and reliable measure of the thymic function. TRECs are a by-product of DNA formation in gene rearrangement of T cell receptors. They are stable and they do not duplicate during mitosis, representing the recent emigrant T cells from the thymus. Despite their importance, TRECs have been neglected by physicians and there is a lack of data regarding thymic function during infancy of healthy children. In order to evaluate thymic function in the first years of life, we propose measuring TRECs as a valuable tool. One hundred and three blood samples from children and adolescents between 3 months and 20 years of age were analyzed. The mean TRECs count was 136.77±96.7 copies of TRECs/μL of DNA. The individuals between 0 and 5 years of age had significantly higher TRECs values than those between 10 and 20 years of age. No significant difference was observed in TRECs values among age groups below 5 years of age. An inverse correlation between TRECs and age was found (r=0.3 P=0.003). These data highlight and validate the evidence of decreased thymus function with age, even during infancy. Awareness should be raised with this important albeit ignored organ.

Hipotiroidismo subclínico en mujeres infértiles / Subclinical Hypothyroidism in Infertile Women

Objetivo: Comparar la frecuencia de hipotiroidismo subclínico (HSC) entre mujeres con infertilidad primaria (IP) y secundaria (IS) asistidas en la consulta de ginecología de la Maternidad “Dr. Armando Castillo Plaza”, en Maracaibo, estado Zulia. Métodos: Investigación comparativa y aplicada, con diseño de tipo no experimental, contemporáneo transeccional y de campo en la cual se incluyeron 100 mujeres con infertilidad, separadas en dos grupos pareados en relación con el tipo de infertilidad, a las cuales se les determinó el perfil hormonal tiroideo: hormona estimuladora del tiroides (TSH), tiroxina libre (FT4) y triiodotiroxina libre (FT3). Resultados: No se evidenciaron diferencias estadísticas significativas entre las mujeres con IP o IS, en cuanto a edad, menarquía, sexarquía, número de parejas y tiempo de convivencia. En las mujeres con IP prevaleció el factor ovulatorio (50 %) seguido del tubo-peritoneal (34 %), mientras que entre las pacientes con IS el factor principal fue el tubo-peritoneal (38 %) seguido del ovulatorio (28 %); mostrando solamente el factor ovulatorio una diferencia significativa (p< 0,05). Se determinó que las pacientes con IP presentaron significativamente concentraciones más elevadas tanto de la TSH (3,14 ± 2,08 vs. 2,33 ± 1,59; p< 0,05) como de FT4 (1,19 ± 0,37 vs. 1,04 ± 0,25; p< 0,05); determinándose una prevalencia de HSC en mujeres infértiles del 17 %; siendo más frecuente y significativo en el grupo de mujeres con IP que en las pacientes con IS (26 % vs. 8 %; OR [IC95%]= 4,04 [1,26 -13,43]; p < 0,05), en tanto que según la severidad del HSC todos los casos detectados en ambos grupos se encontraban dentro del grado I. Conclusión: Existe una mayor prevalencia de HSC entre las mujeres con IP que en las pacientes con IS.

Objective: To compare the frequency of subclinical hypothyroidism (SCH) among women with primary (PI) and secondary infertility (SI) who presented for gynecology consultation at Maternity "Dr. Armando Castillo Plaza" in Maracaibo, Zulia state. Methods: Comparative and applied research with non-experimental, contemporary transactional and field design, including 100 women with infertility, separated into two groups matched for the type of infertility. Thyroid hormone profile: thyroid stimulating hormone (TSH), free thyroxine (FT4) and free triiodothyroxine (FT3) were measured in these women. Results: There were no statistically significant differences between women with PI or secondary SI, in terms of age, menarche, sexarche, number of partners and length of cohabitation. In women with PI the ovulatory factor prevailed (50 %) followed by the tube-peritoneal (34 %), while among patients with SI the main factor was the tube-peritoneal (38 %) followed by the ovulatory (28 %); only the ovulatory factor showed a significant difference (p < 0.05). PI patients had significantly higher concentrations of both TSH (3.14 ± 2.08 vs. 2.33 ± 1.59, p <0.05) and FT4 (1.19 ± 0, 37 vs. 1.04 ± 0.25, p < 0.05); the prevalence of SCH was determined at 17% in infertile women, being more frequent and significant in the PI group of women than in SI patients (26 % vs. 8 %, OR [95 % CI] = 4.04 [1.26-13.43], p < 0.05), while according to the severity of SCH all cases detected in both groups were within grade I. Conclusion: There is a higher prevalence of SCH in women with PI than in patients with SI.

Evaluation of the hypoglycaemic activity of Petiveria alliacea (Guinea Hen Weed) extracts in normoglycaemic and diabetic rat models / Evaluación de la actividad hipoglicémica de los extractos de Petiveria alliacea (Yerba de Guinea o Anamú) en modelos de ratas diabéticas y normoglicémicase

OBJECTIVE: Petiveria alliacea (p alliacea) has ethno-traditional use as a hypoglycaemic agent in Jamaica and is yet to be scientifically validated as such. Therefore, extracts of aerial parts of the plant were evaluated for hypoglycaemic activity in normoglycaemic and diabetic rats. METHODS: Aqueous and hexane extracts prepared from leaves of p alliacea were tested for hypoglycaemic activity. An acute administration of the extracts (200 and 400 mg/kg body weight) was evaluated in normoglycaemic rats. Additionally, the hypoglycaemic effect ofsub-chronic administration was assessed in streptozotocin-induced diabetic rats. Blood glucose was recorded using a glucometer and test strips. Data were analysed using Student's t-test (p < 0.05). RESULTS: The aqueous and hexane extracts demonstrated no significant reduction of fasting blood glucose (FBG) and no significant improvement of glucose tolerance in normal rats. The aqueous extract (400 mg/kg body weight) increased FBG from 4.75 ± 0.28 mmol/L to 5.88 ± 0.46 when compared to control (p < 0.001). In diabetic rats, the hexane extract (400 mg/kg body weight) caused reduction of FBG after two weeks of treatment (p < 0.010), but this was not sustained. The aqueous extract showed no reduction of FBG in diabetic rats. CONCLUSION: The aqueous extract of p alliacea demonstrated a hyperglycaemic effect in normoglycaemic rats and showed no hypoglycaemic activity in diabetic rats. The hexane extract caused no hypoglycaemic action in normal rats and failed to sustain an initial hypoglycaemic action in diabetic rats. This study presents evidence that does not support significant hypoglycaemic activity of p alliacea; this could hold significant implications for its use in ethno-traditional medicine.

OBJETIVO: Petiveria alliacea (p alliacea) tiene uso etnotradicional como agente hipoglicémico en Jamaica, y todavía requiere ser validado científicamente. Por lo tanto, extractos de las partes aéreas de la planta fueron evaluados en relación con su actividad hipoglicémica en ratas normoglicémicas y diabéticas. MÉTODOS: Extractos acuosos y extractos de hexanos preparados a partir de hojas de p alliacea fueron sometidos a prueba a fin de detectar su actividad hipoglicémica. Se evaluó el efecto de una administración aguda de los extractos (200 y 400 mg/kg de peso corporal) en ratas normoglicémicas. Además, se evaluó el efecto hipoglicémico de la administración subcrónica en ratas con diabetes inducida por estreptozotocina. La glucosa en sangre fue registrada usando un glucómetro y tiras reactivas. Los datos se analizaron mediante la prueba t de Student (p < 0.05). RESULTADOS: Los extractos acuosos y los extractos de hexano no mostraron reducción significativa alguna de la glucemia en ayunas (GA), ni tampoco ninguna mejora significativa de la tolerancia a la glucosa en ratas normales. El extracto acuoso (400 mg/kg de peso corporal) aumentó la GA de 4,75 ± 0,28 mmol/L a 5,88 ± 0,46 en comparación con el control (p < 0.001). En las ratas diabéticas, el extracto de hexano (400 mg/kg de peso corporal), trajo por consecuencia la reducción de GA tras dos semanas de tratamiento (p < 0.010), pero este efecto no se mantiene. El extracto acuoso no mostró ninguna reducción de GA en las ratas diabéticas. CONCLUSIÓN: El extracto acuoso de p alliacea mostró un efecto hiperglicémico en las ratas normoglicémicas, y no mostró ninguna actividad hipoglicémica en las ratas diabéticas. El extracto de hexano no produjo ninguna acción hipoglicémica en ratas normales, y no mantuvo la acción hipoglicémica inicial en las ratas diabéticas. Este estudio presenta evidencias que no respaldan una actividad hipoglicémica significativa de p alliacea, lo cual podría tener importantes implicaciones para su uso en la medicina etnotradicional.

Cassia alata leaf extract induces cytotoxicity in A549 lung cancer cells via a mechanism that is caspase 8 dependent / El extracto de la hoja de cassia alata induce citotoxicidad en las células A549 del cáncer pulmonar humano a través de un mecanismo dependiente de la caspasa 8

OBJECTIVE: To evaluate the cytotoxic effect of a hexane extract of Cassia alata leaves in A549 lung cancer cells. METHOD: Parental A549 lung cancer cells were exposed to various concentrations (100"180 µg/ml) of Cassia alata leaf extract for 24 hours. Following treatment, the cells were evaluated using the 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay to determine the cytotoxic effect of the extract. Caspase 8, 3 and 9 negative A549 cells were also prepared using lentiviral based shRNA knockdown of the caspase 8, 3 and 9 genes, respectively. The cytotoxic effect of Cassia alata leaf extract was then evaluated in these knockdown cells using the MTT assay. Chemical analysis was performed on the extract using high performance liquid chromatography (HPLC). RESULTS: Cassia alata extract was cytotoxic in parental and caspase-9 negative, but not caspase 3 and 8 negative A549 cells. The IC50 values were 143 µg/ml and 145 µg/ml in parental and caspase 9 negative A549 cells respectively. The flavanoid kaempferol was identified as a constituent of Cassia alata leaf extract. CONCLUSIONS: Cassia alata produces cytotoxicity in A549 cancer cells that is mediated by caspase 8 activation. This effect may be attributable to kaempferol.

OBJETIVO: Evaluar el efecto citotóxico de un extracto de hexano de hojas de Cassia alata en las células A549 del cáncer pulmonar. MÉTODO: Células A549 parentales del cáncer pulmonar fueron expuestas a varias concentraciones (100-180 µg/ml) de un extracto de la hoja de Cassia alatadurante 24 horas. Tras el tratamiento, las células fueron evaluadas usando el ensayo de bromuro de 3-(4,5-dimetiltiazol-2-il)-2,5-difeniltetrazolio (MTT) a fin de determinar el efecto citotóxico del extracto. También se prepararon células A549 negativas caspasa 8, 3 y 9 mediante silenciamiento génico vía ARN (shRNA knockdown) de los genes de las caspasas 8, 3 y 9 respectivamente, sobre la base de la inserción de vectores lentivirales. Entonces, usando un ensayo MTT se procedió a evaluar el efecto citotóxico del extracto de hojas de Cassia alataen éstas células genéticamente modificadas. Se realizó un análisis químico del extracto utilizando cromatografía líquida de alta eficacia. (HPLC). RESULTADOS: El extracto de Cassia alata resultó ser citotóxico en las células A549 negativas parentales y caspasa 9, pero no en las negativas caspasa 3 y 8. Los valores de IC50 fueron 143 µg/ml y 145 µg/ml en las células A549 negativas parentales y caspasa 9 respectivamente. El flavonol kaempferol fue identificado como un constituyente del extracto de las hojas de Cassia alata. CONCLUSIONES: La Cassia alata produce citotoxicidad en las células cancerosas A549, mediada por la activación de la caspasa 8. Este efecto puede ser atribuido al kaempferol.

Anti-inflammatory activities of cassia alata leaf extract in complete freund's adjuvant arthritis in rats / Actividades anti-inflamatorias del extracto de hojas de cassia alata en artritis inducida en ratas mediante adyuvante completo de freund

OBJECTIVE: To investigate the anti-inflammatory effects of a hexane extract of Cassia alata leaves in complete Freund's adjuvant (CFA) arthritis in rats. METHOD: A hexane extract of Cassia alata leaves was administered by oral gavage to CFA arthritic rats (500 mg/kg, n = 6). Controls received corn oil (2 ml, n = 6). The CFA arthritic model was induced by the injection of 0.5 ml (CFA) into the synovial cavity of the right knee joint of the hind leg of rats. The ability of the plant extract to reduce swelling as a sign of arthritic inflammation was assessed by obtaining the circumference of the knee joint before and for twenty eight days post arthritis induction. Reduction of leukocyte infiltration into the blood and synovial cavity of the arthritic rats were assessed using automated counting and Wrights method. Protection against cartilage erosion was also assessed histologically. RESULTS: Cassia alata extract significantly (p = 0.0032) reduced knee circumference (swelling) in the CFA arthritic rats. Total and differential leukocyte counts in both blood and synovial fluid from Cassia alata treated animals were significantly (p < 0.05) lower than in control animals. Protective effects against cartilage degradation on the femoral head of the knee joint were observed in Cassia alata treated animals, as normal cartilage structure and chondrocyte arrangement were maintained. CONCLUSIONS: The results suggest that Cassia alata exhibits anti-inflammatory activities that should be further examined and potentially exploited for anti-arthritic therapies.

OBJETIVO: Investigar los efectos anti-inflamatorios del extracto de hexano de hojas de Cassia alata en artritis inducida por adyuvante completo de Freund (CFA) en ratas. MÉTODO: Un extracto de hexano de hojas de Cassia alata fue administrado por gavage oral a ratas artríticas por CFA (500 mg/kg, n = 6). Los controles recibieron aceite de maíz (2 ml, n = 6). El modelo artrítico de CFA fue inducido inyectando 0.5 ml (CFA) en la cavidad sinovial de la rótula derecha de la pata trasera de las ratas. La capacidad del extracto de la planta en cuanto a reducir la inflamación como signo de la inflamación artrítica, fue evaluada obteniendo la circunferencia de la rótula antes y durante veintiocho días posterior a la inducción de la artritis. La reducción de la infiltración de leucocitos en la sangre y la cavidad sinovial de las ratas artríticas fue evaluada usando el conteo automatizado y el método de Wright. También se evaluó histológicamente la protección contra la erosión del cartílago. RESULTADOS: El extracto de Cassia alata redujo significativamente (p = 0.0032) la circunferencia de la rodilla (inflamación) en las ratas artríticas por CFA. Los conteos totales y diferenciales de leucocitos tanto en la sangre como en el líquido sinovial de los animales tratados con Cassia alata fueron significativamente (p < 0.05) más bajos en los animales del control. Los efectos protectores contra la degradación del cartílago en la cabeza femoral de la rótula fueron observados en los animales tratados con Cassia alata, ya que se mantuvieron la estructura normal del cartílago y las disposición de los condrocitos. CONCLUSIONES: Los resultados sugieren que la Cassia alata exhibe propiedades anti-inflamatorias que deben ser examinadas ulteriormente y explotadas potencialmente para las terapias anti-artríticas.

Cardiovascular disease prevention in Ghana: feasibility of a faith-based organizational approach

Objective To examine the feasibility of using community health workers (CHWs) to implement cardiovascular disease (CVD) prevention programmes within faith-based organizations in Accra; Ghana. Methods Faith-based organization capacity; human resources; health programme sustainability/barriers and community members' knowledge were evaluated. Data on these aspects were gathered through a mixed method design consisting of in-depth interviews and focus groups with 25 church leaders and health committee members from five churches; and of a survey of 167 adult congregants from two churches. Findings The delivery of a CVD prevention programme in faith-based organizations by CHWs is feasible. Many faith-based organizations already provide health programmes for congregants and involve non-health professionals in their health-care activities; and most congregants have a basic knowledge of CVD. Yet despite the feasibility of the proposed approach to CVD prevention through faith-based organizations; sociocultural and health-care barriers such as poverty; limited human and economic resources and limited access to health care could hinder programme implementation. Conclusion The barriers to implementation identified in this study need to be considered when defining CVD prevention programme policy and planning