Volatilome Analysis of Soursop Fruits for the Determination of Kairomone Components That Attract the Annonaceae Fruit Weevil (Optatus palmaris Pascoe).

Soursop possesses the largest fruit size of the Annona genus. However, this species is threatened by the Annonaceae fruit weevil (Optatus palmaris), which can cause the destruction of whole soursop fruits. Recently, the potential of semiochemicals for the management of this insect is highlighted, and its aggregation pheromone has been elucidated. This pheromone works well only when mixed with soursop volatiles. Thus, the aim of this research was to determine specific kairomone components to potentiate the aggregation pheromone of this Annonaceae fruit weevil. This task was carried out via volatilome analysis of soursop fruits, which was correlated with the biological activity of the identified volatiles. The GC-MS analysis of aroma collections of mature soursop fruits and flowers, determined using multivariate data analysis, confirmed a volatile differentiation between these organs. The volatile variation between fruits and flowers was reflected in weevils' preference for mature fruits instead of flowers. Moreover, weevils' response to soursop fruits increased with more mature fruits. This was correlated with volatile changes throughout the phenological stages of soursop fruits. The two volatiles most correlated with weevils' attraction were benzothiazole and (E)-ß-caryophyllene. These volatiles only evoked a response when mixed and potentiated the attraction of the aggregation pheromone. Thus, these two volatiles are active kairomone components with the potential for being used in combination with the aggregation pheromone of Annonaceae fruit weevils in field trials.

Epidemiology and mortality in patients hospitalized for burns in Catalonia, Spain.

Burn injuries are one of the leading causes of morbidity worldwide. Although the overall incidence of burns and burn-related mortality is declining, these factors have not been analysed in our population for 25 years. The aim of this study has been to determine whether the epidemiological profile of patients hospitalized for burns has changed over the past 25 years. We performed a retrospective cohort study of patients hospitalised between 1 January 2011 and 31 December 2018 with a primary diagnosis of burns. The incidence of burns in our setting was 3.68/105 population. Most patients admitted for burns were men (61%), aged between 35 and 45 years (16.8%), followed by children aged between 0 and 4 years (12.4%). Scalding was the most prevalent mechanism of injury, and the region most frequently affected was the hands. The mean burned total body surface (TBSA) area was 8.3%, and the proportion of severely burned patients was 9.7%. Obesity was the most prevalent comorbidity (39.5%). The median length of stay was 1.8 days. The most frequent in-hospital complications were sepsis (16.6%), acute kidney injury (7.9%), and cardiovascular complications (5.9%). Risk factors for mortality were advanced age, high abbreviated burn severity index score, smoke inhalation, existing cardiovascular disease full-thickness burn, and high percentage of burned TBSA. Overall mortality was 4.3%. Multi-organ failure was the most frequent cause of death, with an incidence of 49.5%. The population has aged over the 25 years since the previous study, and the number of comorbidities has increased. The incidence and severity of burns, and the percentage of burned TBSA have all decreased, with scalding being the most prevalent mechanism of injury. The clinical presentation and evolution of burns differs between children and adults. Risk factors for mortality were advanced age, smoke inhalation, existing cardiovascular disease, full-thickness burn, and high percentage of burned TBSA.

Fluid resuscitation management in patients with burns: update.

Since 1968, when Baxter and Shires developed the Parkland formula, little progress has been made in the field of fluid therapy for burn resuscitation, despite advances in haemodynamic monitoring, establishment of the 'goal-directed therapy' concept, and the development of new colloid and crystalloid solutions. Burn patients receive a larger amount of fluids in the first hours than any other trauma patients. Initial resuscitation is based on crystalloids because of the increased capillary permeability occurring during the first 24 h. After that time, some colloids, but not all, are accepted. Since the emergence of the Pharmacovigilance Risk Assessment Committee alert from the European Medicines Agency concerning hydroxyethyl starches, solutions containing this component are not recommended for burns. But the question is: what do we really know about fluid resuscitation in burns? To provide an answer, we carried out a non-systematic review to clarify how to quantify the amount of fluids needed, what the current evidence says about the available solutions, and which solution is the most appropriate for burn patients based on the available knowledge.

Phylogeny of chrysosporia infecting reptiles: proposal of the new family Nannizziopsiaceae and five new species.

We have performed a phenotypic and phylogenetic study of a set of fungi, mostly of veterinary origin, morphologically similar to the Chrysosporium asexual morph of Nannizziopsis vriesii (Onygenales, Eurotiomycetidae, Eurotiomycetes, Ascomycota). The analysis of sequences of the D1-D2 domains of the 28S rDNA, including representatives of the different families of the Onygenales, revealed that N. vriesii and relatives form a distinct lineage within that order, which is proposed as the new family Nannizziopsiaceae. The members of this family show the particular characteristic of causing skin infections in reptiles and producing hyaline, thin- and smooth-walled, small, mostly sessile 1-celled conidia and colonies with a pungent skunk-like odour. The phenotypic and multigene study results, based on ribosomal ITS region, actin and ß-tubulin sequences, demonstrated that some of the fungi included in this study were different from the known species of Nannizziopsis and Chrysosporium and are described here as new. They are N. chlamydospora, N. draconii, N. arthrosporioides, N. pluriseptata and Chrysosporium longisporum. Nannizziopsis chlamydospora is distinguished by producing chlamydospores and by its ability to grow at 5 °C. Nannizziopsis draconii is able to grow on bromocresol purple-milk solids-glucose (BCP-MS-G) agar alkalinizing the medium, is resistant to 0.2 % cycloheximide but does not grow on Sabouraud dextrose agar (SDA) with 3 % NaCl. Nannizziopsis arthrosporioides is characterised by the production of very long arthroconidia. Nannizziopsis pluriseptata produces 1- to 5-celled sessile conidia, alkalinizes the BCP-MS-G agar and grows on SDA supplemented with 5 % NaCl. Chrysosporium longisporum shows long sessile conidia (up to 13 µm) and does not produce lipase.

Epigenetic regulation of PRAME in acute myeloid leukemia is different compared to CD34+ cells from healthy donors: effect of 5-AZA treatment.

PRAME is a tumor associated antigen (TAA) of particular interest since it is widely expressed by lymphoid and myeloid malignancies. Several studies have associated high PRAME RNA levels with good prognosis in acute myeloid leukemia (AML). PRAME expression is regulated at the epigenetic level. For this reason inhibitors of DNA methylation, such as 5-azacytidine, can modulate the expression of this TAAs. In the current study we analyzed the effect of 5-azaC on the expression of PRAME in blasts versus CD34+ cells from healthy donors in an attempt to increase its expression, thus inducing a potential target for therapeutic strategies.

Effect of prostaglandin E1 (PGE1) and sildenafil on serotonin metabolism and some oxidative damage markers in rat prostate gland and brain.

The aim of this study was to evaluate the effect of sildenafil and prostaglandin E1 (PGE1) (drugs used in erectile dysfunction) on production of free radicals in prostate and brain of rat. A single dose of sildenafil (10 mg kg(-1) ) and PGE1 (20 µg kg(-1) ) was given to Sprague-Dawley rats (300 g weight) intraperitoneally. The levels of testosterone were measured in blood. Their brains and prostate glands were separated to measure lipid peroxidation, Na(+) and K(+) ATPase activity, reduced glutathione (GSH) and serotonin levels, by means of validated methods. The levels of testosterone increased slightly in animals treated with sildenafil and PGE1. The activity of total ATPase was increased in the prostate of animals treated with sildenafil + PGE1 but decreased in those that received sildenafil alone. PGE1 caused significant diminution of GSH levels in both organs. Sildenafil increased the levels of serotonine in brain, whereas in prostate they decreased instead. Our results suggest that sildenafil induced changes in GSH levels as well as in the serotonergic metabolism, alone or with PGE1 in prostate and brain, respectively. Thus, the combination therapy may be ideal to sustain the biochemical balance due to biphasic stimulation on brain and prostate.

Effect of orlistat on lipid peroxidation, Na+, K+ ATPase, glutathione and serotonin in rat brain.

The aim of this study was to evaluate the effect of orlistat, a drug used in weight loss, on 5-HT and indicators of oxidative stress in rat brain. Orlistat, 12 mg/kg was administered to Wistar rats as single dose or successive doses on 3 consecutive days. Blood glucose and oxidative stress indicators were detected by measurement of lipid peroxidation, Na+, K+ ATPase, glutathione and serotonin levels using previous validated methods. The levels of glucose decreased in rats receiving successive doses. The activity of Na+, K+ ATPase and total ATPase was reduced in rats receiving successive doses, while the level of lipid peroxidation increased slightly in both groups. Glutathione underwent significant reduction in the successive doses group (p < 0.05). 5-HT increased significantly after single dose treatment (p < 0.05). Orlistat can induce pro-oxidant effects in the brain due to alteration of serotonergic metabolism and the reduction of glutathione.

p22phox-dependent NADPH oxidase activity is required for megakaryocytic differentiation.

Transient reactive oxygen species (ROS) production is currently proving to be an important mechanism in the regulation of intracellular signalling, but reports showing the involvement of ROS in important biological processes, such as cell differentiation, are scarce. In this study, we show for the first time that ROS production is required for megakaryocytic differentiation in K562 and HEL cell lines and also in human CD34(+) cells. ROS production is transiently activated during megakaryocytic differentiation, and such production is abolished by the addition of different antioxidants (such as N-acetyl cysteine, trolox, quercetin) or the nicotinamide adenine dinucleotide phosphate (NADPH) oxidase inhibitor diphenylene iodonium. The inhibition of ROS formation hinders differentiation. RNA interference experiments have shown that a p22(phox)-dependent NADPH oxidase activity is responsible for ROS production. In addition, the activation of ERK, AKT and JAK2 is required for differentiation, but the activation of phosphatidylinositol 3-kinase and c-Jun N-terminal kinase seems to be less important. When ROS production is prevented, the activation of these signalling pathways is partly inhibited. Taken together, these results show that NADPH oxidase ROS production is essential for complete activation of the main signalling pathways involved in megakaryocytopoiesis to occur. We suggest that this might also be important for in vivo megakaryocytopoiesis.

In leukapheresis products from non-Hodgkin's lymphoma patients, the immature hematopoietic progenitors show higher CD90 and CD34 antigenic expression.

Damage to the stem cell progenitors caused by the chemotherapy received in patients diagnosed with non-Hodgkin's lymphoma (NHL) may be an important factor limiting progenitor cell mobilization. The aim of the present analysis was to evaluate the effect of the chemotherapy on the different progenitor cell subpopulations obtained in the leukapheresis. For this purpose, a combination of immunophenotype and functional assays has been performed in 26 mobilized peripheral blood (PB) samples from NHL patients and 36 healthy donors. The different progenitor subpopulations analyzed by flow cytometry significantly correlated with the corresponding populations assessed by functional assays in both healthy donors and NHL patients (p<0.05, r>0.5). The number of committed CFU-GM was similar in both groups (p=0.246), but we found significant decrease in the number of BFU-E and more immature progenitors in PB from NHL patients as compared to donors (p<0.05). Moreover, the number of total CFU was significantly lower in NHL patients (p=0.007). Accordingly, CD34+ cells (p=0.018) and CD34+ subpopulations was decreased in NHL patients. Nevertheless, CD90 and CD34 intensity was significantly higher within CD34+ cells from NHL patients as compared to donors. However, although numerically reduced non-committed CD34+ cells are more immature in chemotherapy mobilized NHL patients. In summary, our results show that all NHL hematopoietic progenitors, analyzed by both immunophenotypical and functional approaches, are impaired in leukapheresis products.

Dermatophytosis caused by Trichophyton mentagrophytes in the southern chamois (Rupicapra pyrenaica) in the Eastern Pyrenees.

Two free-ranging southern chamois (Rupicapra pyrenaica) - one 4-year-old male and one 3-year-old male - were diagnosed with clinical dermatophytosis in the French and Spanish Eastern Pyrenees respectively. Skin samples were collected for microbiological studies, Trichophyton mentagrophytes being isolated and identified in both animals. The first chamois was found dead at the base of a cliff, and presented with alopecia and scaling on the dorsum and left forelimb. The second chamois showed grey-yellow, rough to raised scaling and crusting skin lesions at the base of the horns, around the eyes, dorsum of the nose, sternum, tail and limbs. Histological examination was carried out only on the second animal. The main lesions were orthokeratotic hyperkeratosis with focal parakeratosis, irregular to papillary epidermal hyperplasia, intracorneal neutrophilic pustules, perivascular to diffuse dermatitis with neutrophilic folliculitis and furunculosis. In those follicles with folliculitis and/or furunculosis, fungal hyphae and arthrospores associated with the follicular keratin and hair shafts were seen.

Estudio longitudinal de los síntomas de estreñimiento y hábitos dietéticos en la población infantil. Estudio FREI / Longitudinal study of constipation symptoms and dietetic habits in children. FREI study

El objetivo de este estudio es el seguimiento clínico y nutricional de los niños con problemas de estreñimiento, tras informar a sus padres sobre las medidas higiénico-dietéticas a seguir para la corrección del mismo. Mediante un diseño prospectivo y unicéntrico, se recogieron datos clínicos basales y de seguimiento de 307 niños. Se utilizó una encuesta nutricional cada 3 meses durante un período de observación de 6 meses. El número de deposiciones durante el período de seguimiento aumentó significativamente. La incidencia de rectorragia disminuyó de forma estadísticamente significativa así como la necesidad de enemas. Entre el 93 y 100 por ciento de los niños reportaron que consumían lácteos durante el desayuno, observándose un relativo descenso en la ingesta de repostería a lo largo del estudio. El número de pacientes que refirieron consumir fruta durante la comida aumentó tras la intervención, mientras que en la cena se observó un aumento gradual del consumo de verduras y frutas. La actuación precoz en la modificación de hábitos dietéticos mejora los síntomas del estreñimiento infantil. Es necesario el seguimiento posterior para procurar mantener la adhesión a estos hábitos por parte de los niños y de sus progenitores. (AU)

Proliferation of C6 glioma cells is blunted by the increase in gap junction communication caused by tolbutamide.

We have previously reported that tolbutamide prevents the inhibition of gap junction communication in astrocytes. Here, we show that tolbutamide increases gap junction communication and connexin 43 expression in poorly coupled C6 glioma cells. The increase in communication is concurrent with the inhibition of the rate of proliferation due to a block of the progression of C6 glioma cells through the S phase of the cell cycle. The effects of tolbutamide were quantitatively similar to that found after the elevation of intracellular cAMP. Furthermore, the effects of tolbutamide and cAMP were additive. The possible beneficial effect of tolbutamide on gene therapy for gliomas is discussed.

Oligodendrocytes use lactate as a source of energy and as a precursor of lipids.

Lactate is an important metabolic substrate for the brain during the postnatal period and also plays a crucial role in the traffic of metabolites between astrocytes and neurons. However, to date there are no clues with regard to lactate utilization by oligodendrocytes, the myelin-forming cells in the brain. In the present work, lactate utilization by oligodendrocytes in culture was investigated and compared with its utilization by cultured neurons, type 1 and type 2 astrocytes. Our results clearly indicate that oligodendrocytes readily use lactate both as a metabolic fuel and as a precursor to build carbon skeletons. Oligodendrocytes oxidize lactate at a higher rate than that observed for neurons and astrocytes, and their rate of lipid synthesis from lactate was at least 6-fold higher than that found in astrocytes or neurons. The rate of glucose utilization through different pathways was also investigated. The flux of glucose through the pentose phosphate pathway and the rate of lipid synthesis were at least 2-fold higher in oligodendrocytes than in astrocytes or neurons. These findings indicate that oligodendrocyte metabolism is designed specifically for the synthesis of lipids, presumably those of myelin.

Evaluation of a CD61 MoAb method for enumeration of platelets in thrombocytopenic patients and its impact on the transfusion decision-making process.

BACKGROUND: Almost all automated hematology cell analyzers use methods based on either the impedance (PLTi) or the optical (PLTo) properties of the cells for performing platelet counts. To improve the accuracy of platelet counts in peripheral blood (PB), the use of CD61 (GPIIIa) MoAbs (ImmunoPLT method) has recently been introduced in an automated hematology blood-analyzer system (Cell-Dyn 4000, Abbott Diagnostics). STUDY DESIGN AND METHODS: A comparative evaluation was made of the accuracy and precision of the three methods currently available in the Cell-Dyn 4000 automated hematology cell analyzer for counting the number of platelets per microliter of PB in a total of 47 patients with chemotherapy-induced thrombocytopenia. A flow cytometric PB platelet count was also performed in parallel and used as an external reference. RESULTS: PB platelet counts showed a good correlation among the PLTo, CD61-ImmunoPLT, and flow cytometric methods. In contrast, the PLTi procedure usually provided an overestimation of the number of platelets per microliter. Although a good correlation was observed between the flow cytometric reference method and both the ImmunoPLT and PLTo methods, the highest degree of agreement was found for the ImmunoPLT techniques (94% vs. 67%). A comparative analysis of the PLTo and CD61-ImmunoPLT methods with regard to their value for predicting platelet transfusion needs on the basis of specific flow cytometric platelet count thresholds showed a good correlation when the cutoff level of 10,000 platelets per microL was used. In contrast, at the threshold of 20,000 platelets per microL, slight differences were observed between the PLTo and CD61-ImmunoPLT procedures for predicting transfusion needs. CONCLUSION: Such results indicate that, if the CD61-ImmunoPLT method is used in the platelet transfusion decision-making process, unnecessary platelet transfusions could be avoided in up to 17.5 percent of persons with a PLTo count of <20,000 platelets per microL.

The effect of albumin on astrocyte energy metabolism is not brought about through the control of cytosolic Ca2+ concentrations but by free-fatty acid sequestration.

Albumin is an important serum protein that under normal circumstances is not present in the brain. However, during development, under hypoxia, or after breakdown of the blood-brain barrier, albumin is found in the brain, where it is able to regulate energy metabolism. In this work the mechanism through which albumin regulates astrocyte metabolism was investigated. Our results show that albumin strongly increases (more than 100%) the flux of glucose and lactate through the pyruvate dehydrogenase-catalyzed reaction in astrocytes from primary culture. However, albumin only slightly stimulated other metabolic pathways, such as the tricarboxylic acid cycle or the pentose phosphate shunt, indicating that it exerts its effect specifically on the reaction catalyzed by pyruvate dehydrogenase. Although albumin increased cytosolic Ca2+ concentrations in astrocytes, our results show that the increase in pyruvate dehydrogenase activity promoted by albumin is not due to the enhancement of Ca2+ concentrations. Indeed, highly purified albumins failed to increase the Ca2+ concentration but did enhance lactate oxidation. In agreement with this, the effect of albumin on lactate oxidation was not abolished after Ca2+ depletion. Instead, the presence of fatty acids inhibited lactate oxidation and counteracted the effect of albumin, suggesting that albumin activates pyruvate dehydrogenase by binding free fatty acids and/or their CoA-derivatives.

Multicenter evaluation of Neo-Sensitabs, a standardized diffusion method for yeast susceptibility testing.

The agar diffusion method Neo-Sensitabs for sensitivity testing, was evaluated with 33 reference strains by fourteen laboratories. Tablets with 5-fluorocytosine, amphotericin B, nystatin, fluconazole, itraconazole, ketoconazole and tioconazole were used on Shadomy modified medium. These tests classify each strain as susceptible, intermediate or resistant to all tested antifungals by measuring the inhibition zone diameters. Intra and interlaboratory reproducibility was studied. Neo-Sensitabs sensitivity for fungi was easy to perform and reliable method with a reproducibility of 97.1% and superior to other commercialized methods, being specially interesting for antifungal susceptibility in vitro testing of triazole derivatives fluconazole and itraconazole.

Multicenter survey of in vitro antifungal resistance in yeasts of medical importance isolated from Spanish patients.

Twelve Spanish laboratories collected 325 yeast clinical isolates during a 30 day's period, among them 224 Candida albicans, 30 Candida glabrata, and 27 Candida parapsilosis. In vitro antifungal susceptibility to amphotericin B, ketoconazole, fluconazole and itraconazole was determined by an agar diffusion test (Neo-Sensitabs, Rosco, Denmark). All the isolates tested were susceptible in vitroto amphotericin B and nearly all (97.2%) to itraconazole. In vitrosusceptibility to fluconazole and ketoconazole was high (90.2% and 91.4% of isolates, respectively) but showed variations depending on the species tested. Resistance to fluconazole and ketoconazole was low in C. albicans (4% and 3%, respectively), but 30% of Candida guilliermondii and 36% of C. glabrata isolates were resistant to fluconazole. Ketoconazole resistance was observed in 40% of C. glabrata, and 17% of Candida tropicalis. Resistance to antifungal drugs is very low in Spain and it is related to non-C. albicans isolates.

Report of a case of pyogenic arthritis associated with Actinomyces pyogenes in a chamois (Rupicapra pyrenaica).

A case of pyogenic arthritis associated to an Actinomyces pyogenes infection in a chamois (Rupicapra pyrenaica) is described. The animal was a 5-year-old female that had been seen alone in a wooded area close to a small town in November 1995, and was lame on one front limb. One month later, it was seen again in a nearby area. Its lameness had worsened and it knuckled on both front limbs while eating. Its nutrition status had worsened considerably. On 8 January 1996 the animal was found dead. It was cachectic, its metatarsal/metacarpal-digital joints were all swollen and its hoofs were longer than normal. The microbiological study of the samples obtained showed a large number of beta-haemolytic colonies subsequently identified as Actinomyces pyogenes. Obligate anaerobic bacteria were not isolated from the samples.

The K-ATP channel regulates the effect of Ca2+ on gap junction permeability in cultured astrocytes.

Using the scrape-loading technique we show that tolbutamide and glybenzcyclamide, two inhibitors of the K+ channel sensitive to ATP (K-ATP channel), partially prevent the inhibition of gap junction permeability promoted by Ca2+ in cultured astrocytes. This effect was dose-dependent, reaching a maximum at 400 microM and 1 microM of tolbutamide and glybenzcyclamide, respectively. The presence of the Ca2+ ionophore A-23187 strongly reduced the concentration of Ca2+ required to block gap junction permeability but did not abolish the effect of tolbutamide and glybenzcyclamide. These results suggest that the effect of these two compounds are not brought about by control of the intracellular concentration of Ca2+ but probably by the promotion of plasma membrane depolarization.