Parasite communities and their ecological implications: comparative approach on three sympatric clupeiform fish populations (Actinopterygii: Clupeiformes), off Rio de Janeiro, Brazil.

Fish parasite communities can be directly influenced by characteristics of host species. However, little is known about the host-parasite relationships in commercially important fish of the southeastern Atlantic. To address this knowledge gap, a comparative analysis of the parasite communities of three sympatric Clupeiformes was conducted. Cetengraulis edentulus (Engraulidae), Opisthonema oglinum (Clupeidae) and Sardinella brasiliensis (Clupeidae) were collected from an estuarine lagoon near Rio de Janeiro, Brazil. Prevalence, abundance and aggregation were estimated for infrapopulations; richness, diversity, evenness and dominance for infracommunities. The three component communities were compared using both quantitative and qualitative components. Canonical discriminant analysis was used to determine if a host population could be characterised by the component community of its parasites. Multivariate models revealed that host species, a proxy for diet and phylogenetic relationships, was the main factor influencing the composition of parasite infracommunities. Diet was found to be the main factor shaping the communities of endoparasites, in which digeneans were dominant and best indicator of host population. Ectoparasites (copepods, isopods and monogeneans) displayed strong host-specificity with some species restricted to a single host population. The similarity of the component communities of the two clupeid populations demonstrated the influence of host phylogeny. Parasite infracommunities exhibited low diversity and high dominance, with many taxa restricted to a single host species (specialists) and few occurring in more than one (generalists). Host phylogeny and by extension, diet, morphology and coevolution with parasites appear to be important factors in determining the host-parasite relationships of clupeiform fish in the southeastern Atlantic.

Early postoperative serum albumin levels as predictors of surgical outcomes in head and neck squamous cell carcinoma.

BACKGROUND: Malnutrition is a common issue in patients with head and neck squamous cell carcinoma and has a negative effect on surgical outcomes. OBJECTIVE: We attempted to determine which malnutrition diagnostic variables can be used as predictors of postoperative complications in patients with head and neck squamous cell carcinoma. METHODS: Forty-one patients undergoing surgery for head and neck squamous cell carcinoma were submitted to a prospective evaluation. Biochemical data, anthropometric measurements and evaluation of body composition were used in the nutritional analysis. RESULTS: Twenty-two patients (53.6%) developed complications. Serum albumin measured on the first postoperative day was the only variable that significantly differed between groups. A cut-off value of 2.8 g/dL distinguished between patients with a complicated and uncomplicated postoperative course. Normalization of albumin levels occurred more frequently and more rapidly in the noncomplicated group. CONCLUSION: Serum albumin measured on the first postoperative day was the only variable that was a predicter of postoperative complications after major head and neck squamous cell carcinoma surgery.

Extraction, purification by cation exchange supermacroporous cryogel and physico-chemical characterization of γ-conglutin from lupin seeds (Lupinus albus L.).

This study focused on the extraction, purification, and physicochemical characterization of γ-conglutin, a protein present in lupin seeds with properties of reducing blood glucose levels. Total protein was extracted with an alkaline-saline solvent, followed by isoelectric precipitation. Chromatographic purification of the precipitated fraction was performed using a cation exchange supermacroporous cryogel column. Electrophoresis of the eluted fraction from chromatography presented a single band of ∼48 kDa under non-reducing conditions (two bands of ∼30 and ∼17 kDa, under reducing conditions) confirming the success of the purification protocol. Liquid chromatography-tandem mass spectrometry analysis confirmed the identity of the protein as γ-conglutin. The purified γ-conglutin had an isoelectric point of 7.51, ß-sheets prevailing as a secondary structure, and denaturation temperature close to 68°C. The outcome of this work showed that γ-conglutin was obtained with a high degree of purity. The proposed purification protocol is simple and can be easily scaled up.

Early postoperative serum albumin levels as predictors of surgical outcomes in head and neck squamous cell carcinoma / Níveis de albumina sérica no pós‐operatório imediato como preditivo de resultados cirúrgicos em carcinoma de células escamosas de cabeça e pescoço

Abstract Background Malnutrition is a common issue in patients with head and neck squamous cell carcinoma and has a negative effect on surgical outcomes. Objective We attempted to determine which malnutrition diagnostic variables can be used as predictors of postoperative complications in patients with head and neck squamous cell carcinoma. Methods Forty-one patients undergoing surgery for head and neck squamous cell carcinoma were submitted to a prospective evaluation. Biochemical data, anthropometric measurements and evaluation of body composition were used in the nutritional analysis. Results Twenty-two patients (53.6%) developed complications. Serum albumin measured on the first postoperative day was the only variable that significantly differed between groups. A cut-off value of 2.8 g/dL distinguished between patients with a complicated and uncomplicated postoperative course. Normalization of albumin levels occurred more frequently and more rapidly in the noncomplicated group. Conclusion Serum albumin measured on the first postoperative day was the only variable that was a predicter of postoperative complications after major head and neck squamous cell carcinoma surgery.

Resumo Introdução A desnutrição é um problema comum em pacientes com carcinoma de células escamosas de cabeça e pescoço e tem um efeito negativo nos resultados cirúrgicos. Objetivo Tentamos determinar quais variáveis diagnósticas de desnutrição podem ser usadas como preditivos de complicações pós‐operatórias em pacientes com carcinoma de células escamosas de cabeça e pescoço. Método Quarenta e um pacientes submetidos à cirurgia de carcinoma de células escamosas de cabeça e pescoço foram submetidos a uma avaliação prospectiva. Dados bioquímicos, medidas antropométricas e avaliação da composição corporal foram usados na análise nutricional. Resultados Vinte e dois pacientes (53,6%) desenvolveram complicações. A dosagem de albumina sérica no primeiro dia pós‐operatório foi a única variável que diferiu significantemente entre os grupos. Um valor de corte de 2,8 g/dL distinguiu os pacientes com uma evolução pós‐operatória complicada e não complicada. A normalização dos níveis de albumina ocorreu mais frequentemente e mais rapidamente no grupo sem complicação pós‐operatória. Conclusão A albumina sérica medida no primeiro dia pós‐operatório foi a única variável capaz de predizer complicações pós‐operatórias após cirurgia de carcinoma de células escamosas de cabeça e pescoço de grande porte.

Síndrome congénito de Klippel-Trenaunay-Weber: caso clínico / Congenital Klippel-Trenaunay-Weber syndrome: clinical case

RESUMEN: El síndrome de Klippel-Trenaunay-Weber (SKTW) es una enfermedad congénita poco frecuente caracterizada por hipertrofia de un miembro que afecta a los huesos y a las partes blandas, con extensos angiomas planos, várices y otras anomalías vasculares, como fístulas arteriovenosas. Su incidencia es de 1:100.000 personas. El objetivo fue describir un caso raro de SKTW bilateral, confirmado con hallazgos clínicos e imagenológicos en un niño de 9 años de edad, de sexo masculino, con antecedentes de sangrado digestivo bajo, herniorrafía inguinal izquierda y orquidopexia ipsilateral. En ambos miembros inferiores se identificaron lesiones hemangiomatosas e hipertrofia muscular y edema en miembro inferior izquierdo con ausencia de segundo y tercer dedos del pie izquierdo por antecedente quirúrgico de amputación. Adicionalmente, presentaba adenopatías cervicales e inguinales. Dentro de los hallazgos radiográficos importantes, se observó una cortical ósea delgada en el fémur del miembro inferior izquierdo. El SKTW afecta típicamente a los miembros inferiores de forma unilateral; este es un caso infrecuente de afección bilateral (con predominio izquierdo). Algunos pacientes, registran compromiso visceral con hemorragia digestiva baja, además de alteraciones genitourinarias, presentes en el 30% de los casos. La presentación de adenopatías es inusual.

SUMMARY: Klippel-Trenaunay-Weber syndrome (KTWS) is a rare congenital disease characterized by limb hypertrophy affecting bones and soft tissues, with extensive flat angiomas, varicose veins, and other vascular abnormalities, such as arteriovenous fistulas. Its incidence is 1: 100,000 people. The aim of this work was to describe a rare case of bilateral KTWS, confirmed with clinical and imaging findings in a 9-year-old male, with a history of lower gastrointestinal bleeding, left inguinal herniorrhaphy and ipsilateral orchidopexy. In both lower limbs, hemangiomatous lesions and muscle hypertrophy were identified, and in the left lower limb edema was identified with absence of the second and third toes due to a surgical history of amputation. Additionally, the patient presented cervical and inguinal lymphadenopathy. Among the important radiographic findings, in the left lower limb femur a thin bony cortex was observed. KTWS typically affects the lower limbs unilaterally; this is an infrequent case of bilateral affection (predominantly left). Some patients have visceral involvement with lower gastrointestinal bleeding, as well as genitourinary alterations, present in 30 % of cases. The presentation of lymphadenopathy is unusual.

Evaluación de la capacidad funcional en escolares con fisiología de corazón univentricular / Assessment of functional capacity among students with univentricular heart

Se realizó un estudio transversal en escolares con corazón univentricular en estadio pos-bypass total de ventrículo derecho con el objetivo de determinar la capacidad funcional basal mediante el test de marcha en 6 minutos e identificar posibles factores determinantes. Participaron 30 pacientes con una mediana de edad de 12 años. Dieciocho pacientes fueron de sexo masculino. La mediana de distancia recorrida fue de 551,3 metros, un 84 % de la distancia teórica para población pediátrica sana. Las variables talla, presión arterial sistólica pretest y saturación arterial de oxígeno de reposo se asociaron significativamente con la distancia recorrida en el modelo de regresión lineal múltiple. No hubo asociación significativa en los metros caminados respecto de las variables sexo, estado nutricional, dignóstico cardiológico inicial, número de cirugías previas y edad al momento del bypass total de ventrículo derecho

A cross-sectional study was done in students with univentricular heart after undergoing total cavopulmonary connection (Fontan procedure) to establish their baseline functional capacity based on the six-minute walk test and identify potential determining factors. Thirty patients were included; their median age was 12 years old. Eighteen patients were males. The median distance walked was 551.3 meters, 84 % of the theoretical distance for a healthy pediatric population. Height, pre-test systolic blood pressure, and resting arterial oxygen saturation showed a significant association with the distance walked in the multiple linear regression model. No significant association was observed in the meters walked in terms of the following outcome measures: sex, nutritional status, baseline cardiological diagnosis, number of prior surgeries, and age at the time of Fontan procedure

Assessment of functional capacity among students with univentricular heart. / Evaluación de la capacidad funcional en escolares con fisiología de corazón univentricular.

A cross-sectional study was done in students with univentricular heart after undergoing total cavopulmonary connection (Fontan procedure) to establish their baseline functional capacity based on the six-minute walk test and identify potential determining factors. Thirty patients were included; their median age was 12 years old. Eighteen patients were males. The median distance walked was 551.3 meters, 84 % of the theoretical distance for a healthy pediatric population. Height, pre-test systolic blood pressure, and resting arterial oxygen saturation showed a significant association with the distance walked in the multiple linear regression model. No significant association was observed in the meters walked in terms of the following outcome measures: sex, nutritional status, baseline cardiological diagnosis, number of prior surgeries, and age at the time of Fontan procedure.

Se realizó un estudio transversal en escolares con corazón univentricular en estadio pos-bypass total de ventrículo derecho con el objetivo de determinar la capacidad funcional basal mediante el test de marcha en 6 minutos e identificar posibles factores determinantes. Participaron 30 pacientes con una mediana de edad de 12 años. Dieciocho pacientes fueron de sexo masculino. La mediana de distancia recorrida fue de 551,3 metros, un 84 % de la distancia teórica para población pediátrica sana. Las variables talla, presión arterial sistólica pretest y saturación arterial de oxígeno de reposo se asociaron significativamente con la distancia recorrida en el modelo de regresión lineal múltiple. No hubo asociación significativa en los metros caminados respecto de las variables sexo, estado nutricional, dignóstico cardiológico inicial, número de cirugías previas y edad al momento del bypass total de ventrículo derecho.

Oestrus ovis in Ecuador: Importance in the Andean sheep farming.

AIM: This study aimed to determine the prevalence of Oestrus ovis in sheep meant for meat commercialization in the main slaughterhouse of the country. MATERIALS AND METHODS: Between October 2015 and December 2015, we assessed the occurrence of Oestrus myiasis in the main slaughterhouse localized in Quito. In total, 80 sheep heads were randomly inspected and necropsied. Larvae were removed from nasal cavities and paranasal sinuses and cleaned. ANOVA (generalized linear model) was used to estimate the relationship between sex, age, and place of origin and presence or absence of parasite larvae. RESULTS: Morphological identification confirmed that 19% (15/80) of the examined animals were positive for Oestrus ovis; from the positive cases, 21% were young animals <12 months old. We found that statistical differences by animal sex, males, were most infested 93% (14/15) than females 7% (1/15). Larvae's L2 were more abundant than other stages (62 of the total 149). 14 of the infested animals were from the Andean places at > 2500 meters above sea level (m.a.s.l.), and only one case from the coastal region at 250 m.a.s.l. with tropical environmental conditions. CONCLUSIONS: Our results showed evidence of the presence of myiasis caused by O. ovis in Andean and coastal places in Ecuador and its adaptation to different environmental conditions from that reported previously in temperate regions from Europe and Africa.

Curcumin-micellar casein multisite interactions elucidated by surface plasmon resonance.

Determine the thermodynamic and kinetic parameters of interaction between micellar casein (MC) and curcumin (CUR) is useful for the application of MC-CUR systems in food products. We used surface plasmon resonance (SPR) and ultraviolet-visible spectrophotometry (UV-vis) to study the complex formation between MC obtained from skimmed milk and CUR, MC carrying capacity, and thermal protection for CUR at a pH of 6.6. An MC could carry about 18,000 molecules of CUR. SPR suggested an enthalpy-driven process (∆H°â€¯= -64.63 kJ∙mol-1 and T∆S° ranging from -42.45 to -44.46 kJ∙mol-1). Temperature increased reduced the rate of MC-CUR complex formation and increased its dissociation rate. The activation energy for the formation of MC-CUR activated complexes was negative for association of free MC and CUR molecules (-62.8 kJ mol-1) and positive for dissociation of the thermodynamically stable complexes (1.80 kJ mol-1). MC protected the CUR against its thermal degradation when it was subjected to different temperatures (30, 40, 50, and 60 °C for 5.5 h). This study shows the importance of characterizing MC-small molecules interactions for better application of MC as a nanocarrier.

Artroplastía parcial de cadera con banda de tensión en fracturas de fémur proximal en pacientes ancianos / Partial hip arthroplasty with tension band in proximal femur fractures in older patients

RESUMEN: Las fracturas intertrocantéricas en adulto mayor presentan gran dificultad para lograr una cicatrización ósea estable y una fijación en la artroplastía, para la recuperación completa y funcional del anciano. En pacientes adultos mayores sin stock óseo, se recomienda realizar anexo a la artroplastía de cadera, la colocación de una banda de tensión, con la finalidad de brindar mayor estabilidad a los fragmentos y sustentación a la artroplastía primaria, con el objetivo de lograr un retorno a la calidad de vida normal. A las cuatro semanas de la cirugía, se evidencia una mejora en la escala funcional alcanzando una puntuación media de 83, considerandose un buen resultado. En los controles subsecuentes se evidencia una media en la escala de Harris de 87. Los pacientes volvieron a su nivel de actividad preoperatoria, más de la mitad de los pacientes fueron capaces de caminar sin apoyos, y finalmente, no se reportaron complicaciones en los controles postquirúrgicos al seguimiento de un año.

SUMMARY: The intertrochanteric fractures in the elderly are very difficult to achieve stable bone healing and fixation in arthroplasty for complete and functional recovery of the elderly. In elderly patients without bone stock, it is recommended to make an attachment to the hip arthroplasty the placement of tension band with the purpose of providing greater stability to the fragments and support to the primary arthroplasty with the aim of a return to the normal quality of life, in the results at 4 weeks post-surgery an improvement in the functional scale is evidenced having as average score 83 taken as good results. In the subsequent controls a mean on the Harris scale of 87 was evidenced, the patients returned to their level of preoperative activity, more than half of the patients were able to walk without support, no complications were reported in the postoperative follow-up controls. of 1 year of patients.

A Rapid Extraction Method for mammalian cell cultures, suitable for quantitative immunoblotting analysis of proteins, including phosphorylated GCN2 and eIF2α.

Many studies require the detection and relative quantitation of proteins from cell culture samples using immunoblotting. Limiting factors are the cost of protease inhibitors, the time required to break cells and generate samples, as well as the high risk of protein loss during cell breakage procedures. In addition, a common problem is the viscosity of lysed samples due to the released genomic DNA. As a consequence, the DNA needs to be broken down prior to denaturing polyacrylamide protein gel electrophoresis (SDS-PAGE), e.g. by passing the sample through a syringe gauge needle, sonication, or DNase treatment. In a quest to find a more cost-effective, fast, and yet robust procedure, we found that cell lysis, protein denaturation, and DNA fragmentation can be done in only two steps: harvesting followed by a simple non-laborious 2nd step. Similarly to many pre-existing cell breakage procedures, in our Rapid Protein Extraction (RPE) method, proteins liberated from cells are immediately exposed to a denaturing environment. However, advantages of our method are: •No breaking buffer is needed, instead proteins are liberated directly into the denaturing protein loading buffer used for SDS-PAGE. Consequently, our RPE method does not require any expensive inhibitors.•The RPE method does not involve post-lysis centrifugation steps; instead all cell material is dissolved during the 2nd step, the mixing-heat-treatment step which is new to this method. This prevents potential protein loss that may occur during centrifugation. In addition, this 2nd step simultaneously shears the genomic DNA, making an additional step for DNA fragmentation unnecessary.•The generated samples are suitable for high-quality quantitative immunoblotting. With our RPE method we successfully quantified the phosphorylated forms of protein kinase GCN2 and its substrate eIF2α. In fact, the western signals were stronger and with less background, as compared to samples generated with a pre-existing method.

Garlic: kinetic drying and thermodynamic properties / Alho: cinética de secagem e propriedades termodinâmicas

Dehydrated garlic is an importance spice in pre-cooked and processed food products, but studies of the drying variables of this product are scarce. Thus, this work aimed to model and study the kinetics of garlic drying at three different temperatures (40, 50 and 60 °C) and analyze the thermodynamic parameters during the process. The drying was analyzed using pre-established mathematical models for agricultural products and equations that describe the thermodynamics of product dehydration. The best mathematical adjustment for all drying temperatures was the Midilli equation. The moisture ratio was seen to decrease during the process for all situations. The effective diffusivity increased with increasing temperature. The enthalpy change was positive during the process and decreased with increasing temperature. The study entropy was negative, indicating a non-spontaneous reaction. In addition, the Gibbs free energy increased with increasing drying temperature. The thermodynamic standards were satisfactory and correctly described the moisture loss by the garlic samples.

O alho desidratado é um tempero de suma importância em produtos pré-cozidos e processados, no entanto, o estudo das variáveis de secagem desse produto é escasso. Diante disso, objetivou-se com esse trabalho estudar a cinética e a modelagem da secagem de alho, em três temperaturas distintas (40, 50 e 60 °C), e analisar os parâmetros termodinâmicos durante o processo. O estudo da secagem foi feito por meio de modelos matemáticos pré-estabelecidos para produtos agrícolas e por equações que descrevem a termodinâmica da desidratação de produtos. O melhor ajuste matemático foi à equação de Midilli para todas as temperaturas de secagem. A razão de umidade apresentou-se decrescente durante o processo para todas as situações. A difusividade efetiva aumentou com o aumento da temperatura. A variação de entalpia apresentou-se positiva durante o processo de secagem e diminui com o aumento da temperatura. A entropia estudada foi negativa, indicando uma reação não-espontânea. E a energia livre de Gibbs aumentou com a elevação da temperatura de secagem. Os padrões termodinâmicos foram satisfatórios e descreveram como ocorreu a perda de umidade pelas amostras de alho.

Protein SUMOylation is Involved in Cell-cycle Progression and Cell Morphology in Giardia lamblia.

The unicellular protozoa Giardia lamblia is a food- and waterborne parasite that causes giardiasis. This illness is manifested as acute and self-limited diarrhea and can evolve to long-term complications. Successful establishment of infection by Giardia trophozoites requires adhesion to host cells and colonization of the small intestine, where parasites multiply by mitotic division. The tight binding of trophozoites to host cells occurs by means of the ventral adhesive disc, a spiral array of microtubules and associated proteins such as giardins. In this work we show that knock down of the Small Ubiquitin-like MOdifier (SUMO) results in less adhesive trophzoites, decreased cell proliferation and deep morphological alterations, including at the ventral disc. Consistent with the reduced proliferation, SUMO knocked-down trophozoites were arrested in G1 and in S phases of the cell cycle. Mass spectrometry analysis of anti-SUMO immunoprecipitates was performed to identify SUMO substrates possibly involved in these events. Among the identified SUMOylation targets, α-tubulin was further validated by Western blot and confirmed to be a SUMO target in Giardia trophozoites.

Osteocondroma escapular: un caso infrecuente / Scapular osteochondroma: an infrequent case

El osteocondroma de la escápula es un tumor beningno muy raro por su ubicación, que produce dolor y disfunción mecánica de la articulación cuando se establece en la superficie ventral de la escápula, afecta principalmente a los varones en los períodos del crecimiento óseo, entre 10 y 30 años de edad, representan el 14,4 % de todos los tumores de la escápula y el 49 % de los tumores benignos de la escápula. Surgen en las regiones metafisarias de los huesos largos (fémur, húmero, tibia), causando irritación mecánica que conduce a impotencia funcional, bursitis, e incluso fractura de la escápula, pocos casos en huesos planos han sido descritos. El diagnostico se realiza con la valoración física, clínica y exámenes complementarios, debiendo ser confirmado por el histopatológo previa biopsia. La indicación quirúrgica incluye la disfunción mecánica de una articulación o tendón en relación con el tamaño de la lesión, y del cambio sarcomatoso siendo el procedimiento de elección la escisión quirúrgica abierta. A continuación se describe el caso de una joven de 11 años a quien se diagnosticó un osteocondroma subescapular, previabiopsia escisional.

Osteochondroma of the scapulais a rare benign tumor in view of its location, which produces pain and mechanical dysfunction of the joint when it is on the ventral surface of the scapula, mainly affecting men during bone growth periods, betweenthe ages of 10 and 30, accounting for 14.4 % of allscapula tumors and 49 % of benign tumors of thescapula. They arise in the metaphyseal regions ofthe long bones (femur, humerus, tibia), causing mechanical irritation leading to functional impotence,bursitis, and fracture of the scapula. Few cases inflat bones have been described. Diagnosis is made with physical, clinical and complementary examinations, and must be confirmed by prior histopathology biopsy. The surgical indication includes mechanical dysfunction of a joint or tendon in relationto the size of the lesion, sarcomatous change being the procedure of choice in open surgical excision. The case of an 11-year-old girl who was diagnosed with a subscapular osteochondroma after previous excisional biopsy, is described below.

Perturbations in actin dynamics reconfigure protein complexes that modulate GCN2 activity and promote an eIF2 response.

Genetic and pharmacological interventions in yeast and mammalian cells have suggested a cross-talk between the actin cytoskeleton and protein synthesis. Regulation of the activity of the translation initiation factor 2 (eIF2) is a paramount mechanism for cells to rapidly adjust the rate of protein synthesis and to trigger reprogramming of gene expression in response to internal and external cues. Here, we show that disruption of F-actin in mammalian cells inhibits translation in a GCN2-dependent manner, correlating with increased levels of uncharged tRNA. GCN2 activation increased phosphorylation of its substrate eIF2α and the induction of the integrated stress response master regulator, ATF4. GCN2 activation by latrunculin-B is dependent on GCN1 and inhibited by IMPACT. Our data suggest that GCN2 occurs in two different complexes, GCN2-eEF1A and GCN2-GCN1. Depolymerization of F-actin shifts GCN2 to favor the complex with GCN1, concomitant with GCN1 being released from its binding to IMPACT, which is sequestered by G-actin. These events might further contribute to GCN2 activation. Our findings indicate that GCN2 is an important sensor of the state of the actin cytoskeleton.

IMPACT is a GCN2 inhibitor that limits lifespan in Caenorhabditis elegans.

BACKGROUND: The General Control Nonderepressible 2 (GCN2) kinase is a conserved member of the integrated stress response (ISR) pathway that represses protein translation and helps cells to adapt to conditions of nutrient shortage. As such, GCN2 is required for longevity and stress resistance induced by dietary restriction (DR). IMPACT is an ancient protein that inhibits GCN2. RESULTS: Here, we tested whether IMPACT down-regulation mimics the effects of DR in C. elegans. Knockdown of the C. elegans IMPACT homolog impt-1 activated the ISR pathway and increased lifespan and stress resistance of worms in a gcn-2-dependent manner. Impt-1 knockdown exacerbated DR-induced longevity and required several DR-activated transcription factors to extend lifespan, among them SKN-1 and DAF-16, which were induced during larval development and adulthood, respectively, in response to impt-1 RNAi. CONCLUSIONS: IMPACT inhibits the ISR pathway, thus limiting the activation of stress response factors that are beneficial during aging and required under DR.

The efficacy of Mentha arvensis L. and M. piperita L. essential oils in reducing pathogenic bacteria and maintaining quality characteristics in cashew, guava, mango, and pineapple juices.

This study evaluated the ability of the essential oil from Mentha arvensis L. (MAEO) and M. piperita L. (MPEO) to induce ≥5-log reductions in counts (CFU/mL) of E. coli, L. monocytogenes, and Salmonella enterica serovar Enteritidis in Brain-Heart Infusion broth (BHIB) and cashew, guava, mango, and pineapple juices during refrigerated storage (4±0.5°C). The effects of the incorporation of these essential oils on some physicochemical and sensory parameters of juices were also evaluated. The incorporation of 5, 2.5, 1.25, or 0.625µL/mL of MAEO in BHIB caused a ≥5-log reduction in counts of E. coli and Salmonella Enteritidis after 24h of storage; but only 5µL/mL was able to cause the same reduction in counts of L.monocytogenes. The incorporation of 10µL/mL of MPEO in BHIB caused a ≥5-log reduction in counts of E. coli, Salmonella Enteritidis, and L. monocytogenes after 24h of storage; smaller reductions were observed in BHIB containing 5, 2.5, and 1.25µL/mL of MPEO. Similar reductions were observed when the MAEO or MPEO was incorporated at the same concentrations in mango juice. The incorporation of MAEO or MPEO at all tested concentrations in cashew, guava, and pineapple juices resulted in a ≥5-log reduction in pathogen counts within 1h. The incorporation of MAEO and MPEO (0.625 and 1.25µL/mL, respectively) in fruit juices did not induce alterations in °Brix, pH, and acidity, but negatively affected the taste, aftertaste, and overall acceptance. The use of MAEO or MPEO at low concentrations could constitute an interesting tool to achieve the required 5-log reduction of pathogenic bacteria in cashew, guava, mango, and pineapple fruit juices. However, new methods combining the use of MAEO or MPEO with other technologies are necessary to reduce their negative impacts on specific sensory properties of these juices.

The Gcn2 Regulator Yih1 Interacts with the Cyclin Dependent Kinase Cdc28 and Promotes Cell Cycle Progression through G2/M in Budding Yeast.

The Saccharomyces cerevisiae protein Yih1, when overexpressed, inhibits the eIF2 alpha kinase Gcn2 by competing for Gcn1 binding. However, deletion of YIH1 has no detectable effect on Gcn2 activity, suggesting that Yih1 is not a general inhibitor of Gcn2, and has no phenotypic defect identified so far. Thus, its physiological role is largely unknown. Here, we show that Yih1 is involved in the cell cycle. Yeast lacking Yih1 displays morphological patterns and DNA content indicative of a delay in the G2/M phases of the cell cycle, and this phenotype is independent of Gcn1 and Gcn2. Accordingly, the levels of phosphorylated eIF2α, which show a cell cycle-dependent fluctuation, are not altered in cells devoid of Yih1. We present several lines of evidence indicating that Yih1 is in a complex with Cdc28. Yih1 pulls down endogenous Cdc28 in vivo and this interaction is enhanced when Cdc28 is active, suggesting that Yih1 modulates the function of Cdc28 in specific stages of the cell cycle. We also demonstrate, by Bimolecular Fluorescence Complementation, that endogenous Yih1 and Cdc28 interact with each other, confirming Yih1 as a bona fide Cdc28 binding partner. Amino acid substitutions within helix H2 of the RWD domain of Yih1 enhance Yih1-Cdc28 association. Overexpression of this mutant, but not of wild type Yih1, leads to a phenotype similar to that of YIH1 deletion, supporting the view that Yih1 is involved through Cdc28 in the regulation of the cell cycle. We further show that IMPACT, the mammalian homologue of Yih1, interacts with CDK1, the mammalian counterpart of Cdc28, indicating that the involvement with the cell cycle is conserved. Together, these data provide insights into the cellular function of Yih1/IMPACT, and provide the basis for future studies on the role of this protein in the cell cycle.

Extracellular enolase of Candida albicans is involved in colonization of mammalian intestinal epithelium.

Enolase is secreted by Candida albicans and is present in its biofilms although its extracellular function is unknown. Here we show that extracellular enolase mediates the colonization of small intestine mucosa by C. albicans. Assays using intestinal mucosa disks show that C. albicans adhesion is inhibited, in a dose dependent mode, either by pretreatment of intestinal epithelium mucosa disks with recombinant C. albicans enolase (70% at 0.5 mg/ml enolase) or by pretreatment of C. albicans yeasts with anti-enolase antibodies (48% with 20 µg antiserum). Also using flow cytometry, immunoblots of conditioned media and confocal microscopy we demonstrate that enolase is present in biofilms and that the extracellular enolase is not an artifact due to cell lysis, but must represent functional secretion of a stable form. This is the first direct evidence that C. albicans' extracellular enolase mediates colonization on its primary translocation site. Also, because enolase is encoded by a single locus in C. albicans, its dual role peptide, as glycolytic enzyme and extracellular peptide, is a remarkable example of gene sharing in fungi.

Keeping the eIF2 alpha kinase Gcn2 in check.

The protein kinase Gcn2 is present in virtually all eukaryotes and is of increasing interest due to its involvement in a large array of crucial biological processes. Some of these are universally conserved from yeast to humans, such as coping with nutrient starvation and oxidative stress. In mammals, Gcn2 is important for e.g. long-term memory formation, feeding behaviour and immune system regulation. Gcn2 has been also implicated in diseases such as cancer and Alzheimer's disease. Studies on Gcn2 have been conducted most extensively in Saccharomyces cerevisiae, where the mechanism of its activation by amino acid starvation has been revealed in most detail. Uncharged tRNAs stimulate Gcn2 which subsequently phosphorylates its substrate, eIF2α, leading to reduced global protein synthesis and simultaneously to increased translation of specific mRNAs, e.g. those coding for Gcn4 in yeast and ATF4 in mammals. Both proteins are transcription factors that regulate the expression of a myriad of genes, thereby enabling the cell to initiate a survival response to the initial activating cue. Given that Gcn2 participates in many diverse processes, Gcn2 itself must be tightly controlled. Indeed, Gcn2 is regulated by a vast network of proteins and RNAs, the list of which is still growing. Deciphering molecular mechanisms underlying Gcn2 regulation by effectors and inhibitors is fundamental for understanding how the cell keeps Gcn2 in check ensuring normal organismal function, and how Gcn2-associated diseases may develop or may be treated. This review provides a critical evaluation of the current knowledge on mechanisms controlling Gcn2 activation or activity.