Bioproduction of α-terpineol and R-(+)-limonene derivatives by terpene-tolerant ascomycete fungus as a potential contribution to the citrus value chain.

AIMS: The aims of this article were to select fungal species with high tolerance and high growth rate in mediums supplemented with limonene and citrus essential oils (CEOs), and to test the bioconversion capability of the chosen isolates for the bioproduction of aroma compounds. METHODS AND RESULTS: Based on the use of predictive mycology, 21 of 29 isolates were selected after assaying R-(+)-limonene and CEO tolerance (10 g l-1 ). With a dendrogram divisive coefficient of 0·937, the subcluster two with isolates Aspergillus niger LBM 055, Penicillium sp. LBM 150, Penicillium sp. LBM 151 and Penicillium sp. LBM 154 gathered the highest tolerance and mycelia growth speed. Ultrastructural analysis indicated that culture media containing limonene had no visible toxic activity that could promote morphological changes in the fungal cell wall. The biomass of A. niger LBM055 was distinctive in liquid media supplemented with R-(+)-limonene (0·57 ± 0·07 g) and it was selected to prove bioconversion capacity, under static and agitated conditions, and converted up to 98% of limonene, yielding a wide variety of products that were quantified by GC-FID. It was obtained at molecular weights less than limonene (64-100%), between limonene and α-terpineol (12-72%) and greater than α-terpineol (2-48%). CONCLUSIONS: Aspergillus niger LBM 055, Penicillium sp. LBM 150, Penicillium sp. LBM 151 and Penicillium sp. LBM 154 showed to the highest tolerance and growth rate in mediums supplemented with R-(+)-limonene and orange and lemon essential oils. Particularly, A. niger LBM055, showed limonene bioconversion capability and produced different molecular weights compounds such us α-terpineol. SIGNIFICANCE AND IMPACT OF THE STUDY: Different bioproducts can be obtained by changing operative condition with the same fungus, and this bioprocess aspect is a significant approach to be adopted on industrial scale leading to the creation of new natural flavours and fragrance compositions.

Isolation of a laccase-coding gene from the lignin-degrading fungus Phlebia brevispora BAFC 633 and heterologous expression in Pichia pastoris.

AIMS: Isolate and characterize a laccase-encoding gene (lac I) of Phlebia brevispora BAFC 633, as well as cloning and expressing cDNA of lac I in Pichia pastoris. And to obtain a purified and characterized recombinant laccase to analyse the biotechnological application potential. METHODS AND RESULTS: Lac I was cloned and sequenced, it contains 2447 pb obtained by PCR and long-distance inverse PCR. Upstream of the structural region of the laccase gene, response elements such as metals, antioxidants, copper, nitrogen and heat shock were found. The coding region consisted of a 1563-pb ORF encoding 521 amino acids. Lac I was functionally expressed in P. pastoris and it was shown that the gene cloned using the α-factor signal peptide was more efficient than the native signal sequence, in directing the secretion of the recombinant protein. Km and highest kcat /Km values towards ABTS, followed by 2,6-dimethylphenol, were similar to other laccases. Lac I showed tolerance to NaCl and solvents, and nine synthetic dyes could be degraded to different degrees. CONCLUSIONS: Lac I-encoding gene could be successfully sequenced having cis-acting elements located at the regulatory region. It was found that lac I cDNA expressed in P. pastoris using the α-factor signal peptide was more efficient than the native signal sequence. The purified Lac I exhibited high tolerance towards NaCl and various solvents and degraded some recalcitrant synthetic dyes. SIGNIFICANCE AND IMPACT OF THE STUDY: The cis-acting elements may be involved in the transcriptional regulation of laccase gene expression. These results may provide a further insight into potential ways of optimizing fermentation process and also open new frontiers for engineering strong promoters for laccase production. The Lac I stability in chloride and solvents and broad decolorization of synthetic dyes are important for its use in organic synthesis work and degradation of dyes from textile effluents respectively.

Synthesis, Photocatalytic, and Antifungal Properties of MgO, ZnO and Zn/Mg Oxide Nanoparticles for the Protection of Calcareous Stone Heritage.

More recently, the biological colonization of stone heritage and consequently its biodeterioration has become the focus of numerous studies. Among all microorganisms, fungi are considered to be one of the most important colonizers and biodegraders on stone materials. This is why the development of new antifungal materials requires immediate action. ZnMgO nanoparticles (NPs) have several exciting applications in different areas, highlighting as an efficient antimicrobial agent for medical application. In this research, the application of Zn-doped MgO (Mg1-xZnxO, x = 0.096) NPs obtained by sol-gel method as antifungal coatings on dolomitic and calcitic stones has been explored as a means to develop effective protective coatings for stone heritage. Moreover, the photocatalytic and antifungal activity of Mg1-xZnxO NPs were comparatively studied with single ZnO and MgO NPs. Thus, compared to the MgO and ZnO nanomaterials, the Mg1-xZnxO NPs exhibited an enhanced photocatalytic activity. After UV irradiation for 60 min, 87% methylene blue was degraded over Zn-doped MgO NPs, whereas only 58% and 38% of MB was degraded over ZnO and MgO NPs, respectively. These nanoparticles also displayed a better antifungal activity than that of single pure MgO or ZnO NPs, inhibiting the growth of fungi Aspergillus niger, Penicillium oxalicum, Paraconiothyrium sp., and Pestalotiopsis maculans, which are especially active in the bioweathering of stone. The improved photocatalytic and antifungal properties detected in the Mg1-xZnxO NPs was attributed to the formation of crystal defects by the incorporation of Zn into MgO. The application of the MgO- and Zn-doped MgO NPs as protective coatings on calcareous stones showed important antifungal properties, inhibiting successfully the epilithic and endolithic colonization of A. niger and P. oxalicum in both lithotypes, and indicating a greater antifungal effectiveness on Zn-doped MgO NPs. The use of Zn-doped MgO NPs may thus represent a highly efficient antifungal protection for calcareous stone heritage.

Isolation of the symbiotic fungus of Acromyrmex pubescens and phylogeny of Leucoagaricus gongylophorus from leaf-cutting ants.

Leaf-cutting ants live in an obligate symbiosis with a Leucoagaricus species, a basidiomycete that serves as a food source to the larvae and queen. The aim of this work was to isolate, identify and complete the phylogenetic study of Leucoagaricus gongylophorus species of Acromyrmex pubescens. Macroscopic and microscopic features were used to identify the fungal symbiont of the ants. The ITS1-5.8S-ITS2 region was used as molecular marker for the molecular identification and to evaluate the phylogeny within the Leucoagaricus genus. One fungal symbiont associated with A. pubescens was isolated and identified as L. gongylophorus. The phylogeny of Leucoagaricus obtained using the ITS molecular marker revealed three well established monophyletic groups. It was possible to recognize one clade of Leucoagaricus associated with phylogenetically derived leaf-cutting ants (Acromyrmex and Atta). A second clade of free living forms of Leucoagaricus (non-cultivated), and a third clade of Leucoagaricus associated with phylogenetically basal genera of ants were also recognized. The clades corresponded to traditional taxonomic groups, and were differentiated by ecological habitats of different species.

Metástasis en cóndilo mandibular: presentación de un caso clínico / Mandibular condyle metastasis: report of a clinical case

Las metástasis bucales constituyen un grupo de baja frecuencia pero de gran importancia en particular por su localización. Según datos bibliográficos internacionales, representan del 1 por ciento al 8 por ciento de todas las neoplasias malignas bucales. Pueden localizarse tanto en los tejidos blandos como en los huesos maxilares (84 por ciento) prevaleciendo en la mandíbula, principalmente en el área molar, seguida del sector premolar. La localización condilar es rara. En los últimos 55 años sólo han sido reportados 48 casos. Dado que no existen patrones clínicos ni radiográficos que permitan caracterizar las lesiones, estas patologías suelen en un principio ser tratadas erróneamente como desordenes propios de la articulación temporo mandibular (ATM).Se presenta un caso clínico de a una paciente de sexo femenino de 36 años con antecedentes de adenocarcinoma de recto que presenta una metástasis en cóndilo bajo la apariencia clínica de un cuadro de disfunción de la ATM. Corresponde al tercer caso reportado en la literatura.

Oral metastases are a group of low-frequency lesions but important particularly because of its location. According to international bibliographic data represent 1% to 8% of oral malignancies. They can be located both in soft tissues and in the jaws (84%). Prevails in the mandible, mainly in the molar area, followed by the premolar region. The condylar location is extremely rare. Since the clinical and radiographic features are not characteristic, these lesions are often erroneously treated initially as temporomandibular joint (TMJ) disorders.In the last 55 years only 48 cases have been reported. We present a 36-years-old female patient with antecedents of rectal adenocarcinoma treated two years ago, having a metastasis in the mandibular condyle with clinical appearance of TMJ dysfunction.This is the third case reported in the literature.

Manejo y diagnóstico del incidentaloma suprarrenal / Diagnosis and management of adrenal incidentaloma

La creciente demanda de pruebas radiológicas y el envejecimiento progresivo de la población ha llevado al aumento progresivo en la detección de lesiones suprarrenales indeterminadas. Una vez detectado el incidentaloma suprarrenal, disponemos actualmente de pruebas clínicas y hormonales y de una batería de técnicas de imagen (TC, RM, PET/TC…) que hacen posible el diagnóstico etiológico de la mayoría de estos incidentalomas. En este artículo presentamos las ventajas y limitaciones de cada técnica, las características por imagen de las lesiones suprarrenales más frecuentes y proponemos un algoritmo diagnóstico que permita el diagnóstico de las mismas (AU)

The growing demand for imaging tests and the progressive aging of the population have led to a progressive increase in the detection of indeterminate adrenal lesions. Once an adrenal incidentaloma is detected, clinical and hormone tests together with a battery of imaging techniques (CT, MRI, PET/CT…) make it possible to determine the cause in most cases. In this article, we discuss the advantages and limitations of each technique. We show the imaging characteristics of the most common adrenal lesions and propose a diagnostic algorithm to enable their diagnosis (AU)

[Diagnosis and management of adrenal incidentaloma]. / Manejo y diagnóstico del incidentaloma suprarrenal.

The growing demand for imaging tests and the progressive aging of the population have led to a progressive increase in the detection of indeterminate adrenal lesions. Once an adrenal incidentaloma is detected, clinical and hormone tests together with a battery of imaging techniques (CT, MRI, PET/CT…) make it possible to determine the cause in most cases. In this article, we discuss the advantages and limitations of each technique. We show the imaging characteristics of the most common adrenal lesions and propose a diagnostic algorithm to enable their diagnosis.

Niveles de conocimiento y práctica sobre prevención en salud bucodental de padres cuyos hijos acuden a la Cátedra de Odontopediatría de la Facultad de Odontología de la Universidad Nacional de Asunción en el año 2009 / Levels of knowledge and practice of prevention in oral health of parents whose children attend the Department of Dentistry of the Fucultad of Dentistry, National University of Asunción in 2009

Los padres son los únicos responsables de la salud bucal de sus hijos durante le primeros años de vida, tienen la responsabilidad de inicia a sus hijos en las prácticas de higiene bucal, proporcionarle una dieta equilibrada para favorecer un desarrollo óptimo en todas las estructuras corporal y velar, con la ayuda de un profesional odontológico, por un desarrollo equilibrado de toda la estructura estogmatognática. Este estudio observacional descriptivo de corte transversal, tuvo como objetivo determinar el nivel de conocimiento y práctica sobre prevención en salud bucodental de padres cuyo hijos acudieron a la cátedra de odontopediatría de la Facultad de Odontología de la Universidad Nacional de Asunción durante el año 2009, para lo cual se realizó una encuesta sobre higiene bucal, dieta, Flúor, alimentos cariogénicos y erupción dentaria, que constituyeron las variables de interés. La muestra estuvo constituida por 152 sujetos de ambos sexos. Los resultados del estudio demostraron que un 78,9% de los padres encuestados posee conocimiento suficiente, frente a un 21,19% que posee conocimiento insuficiente

Natural radioactivity in groundwater and estimates of committed effective dose due to water ingestion in the state of Chihuahua (Mexico).

The activity concentration of 222Rn, 226Ra and total uranium in groundwater samples collected from wells distributed throughout the state of Chihuahua has been measured. The values obtained of total uranium activity concentration in groundwater throughout the state run from <0.03 up to 1.34 Bq l-1. Generally, radium activity concentration was <0.16 Bq l-1, with some exceptions; in spring water of San Diego de Alcalá, in contrast, the value reached approximately 5.3 Bq l-1. Radon activity concentration obtained throughout the state was from 1.0 to 39.8 Bq l-1. A linear correlation between uranium and radon dissolved in groundwater of individual wells was observed near Chihuahua City. Committed effective dose estimates for reference individuals were performed, with results as high as 134 microSv for infants in Aldama city. In Aldama and Chihuahua cities the average and many individual wells showed activity concentration values of uranium exceeding the Mexican norm of drinking water quality.

Radon concentrations in ground and drinking water in the state of Chihuahua, Mexico.

This paper reports (222)Rn concentrations in ground and drinking water of nine cities of Chihuahua State, Mexico. Fifty percent of the 114 sampled wells exhibited (222)Rn concentrations exceeding 11Bq/L, the maximum contaminant level (MCL) recommended by the USEPA. Furthermore, around 48% (123 samples) of the tap-water samples taken from 255 dwellings showed radon concentrations over the MCL. There is an apparent correlation between total dissolved solids and radon concentration in ground-water. The high levels of (222)Rn found may be entirely attributed to the nature of aquifer rocks.

Uranium-238 and thorium-232 series concentrations in soil, radon-222 indoor and drinking water concentrations and dose assessment in the city of Aldama, Chihuahua, Mexico.

High-resolution gamma spectrometry was used to determine the concentration of 40K, 238U and 232Th series in soil samples taken from areas surrounding the city of Aldama, in Chihuahua. Results of indoor air short-time sampling, with diffusion barrier charcoal detectors, revealed relatively high indoor radon levels, ranging from 29 to 422 Bq/m3; the radon concentrations detected exceeded 148 Bq/m3 in 76% of the homes tested. Additionally, liquid scintillation counting showed concentrations of radon in drinking water ranging from 4.3 to 42 kBq/m3. The high activity of 238U in soil found in some places may be a result of the uranium milling process performed 20 years ago in the area. High radon concentrations indoor and in water may be explained by assuming the presence of uranium-bearing rocks underneath of the city, similar to a felsic dike located near Aldama. The estimated annual effective dose of gamma radiation from the soil and radon inhalation was 3.83 mSv.

[Floating thrombus in the aortic arch: a rare case of peripheral arterial embolic events (report of a clinical case)]. / Thrombus flottant de la crosse de l'aorte: une cause rare d'embolies artérielles périphériques. A propos d'un cas clinique.

Floating thrombus in the aortic arch is a rare and often under-diagnosed source of peripheral arterial embolic events. We report a case of a patient seen with arterial embolic events: ischemia of the left superior limb and transient stroke. The diagnosis was performed with transesophageal echocardiography and computed tomography. The thrombus completely disappeared after 15 days of oral anticoagulant therapy. Although rare, this diagnosis mustn't be overlooked in the search for an etiology of recurrent and disseminated peripheral ischemic events because the detrimental functional risk related to a delayed diagnosis.

Calibration of diffusion barrier charcoal detectors using a semi-empirical expression.

Several calibration settings of diffusion barrier charcoal canister (DBCC) detectors for measuring radon concentration in air were studied. A set of functions and graphs were developed for relations between radon concentration in air and adsorbed activity in DBCC, when calibrated in small chambers. Both the integration time for 10% of DBCC of a batch, and the radon adsorption coefficient for the activated charcoal used in these detectors, were determined. Thus, a semi-empirical expression for detector calibration was applied.

Phase I trial of continuous infusion flavopiridol, a novel cyclin-dependent kinase inhibitor, in patients with refractory neoplasms.

PURPOSE: We conducted a phase I trial of the cyclin-dependent kinase inhibitor, flavopiridol (National Service Center [NSC] 649890), to determine the maximum-tolerated dose (MTD), toxicity profile, and pharmacology of flavopiridol given as a 72-hour infusion every 2 weeks. PATIENTS AND METHODS: Seventy-six patients with refractory malignancies with prior disease progression were treated with flavopiridol, with first-cycle pharmacokinetic sampling. RESULTS: Forty-nine patients defined our first MTD, 50 mg/m2/d x 3 with dose-limiting toxicity (DLT) of secretory diarrhea at 62.5 mg/kg/d x 3. Subsequent patients received antidiarrheal prophylaxis (ADP) to define a second MTD, 78 mg/m2/d x 3 with DLT of hypotension at 98 mg/m2/d x 3. Other toxicities included a proinflammatory syndrome with alterations in acute-phase reactants, particularly at doses >50 mg/ m2/d x 3, which in some patients prevented chronic therapy every 2 weeks. In some patients, ADP was not successful, requiring dose-deescalation. Although approximately 70% of patients displayed predictable flavopiridol pharmacology, we observed unexpected interpatient variability and postinfusion peaks in approximately 30% of cases. At the two MTDs, we achieved a mean plasma flavopiridol concentration of 271 nM (50 mg/m2/d x 3) and 344 nM (78 mg/m2/d x 3), respectively. One partial response in a patient with renal cancer and minor responses (n=3) in patients with non-Hodgkin's lymphoma, colon, and renal cancer occurred. CONCLUSION: The MTD of infusional flavopiridol is 50 mg/m2/d x 3 with dose-limiting secretory diarrhea at 62.5 mg/m2/d x 3. With ADP, 78 mg/m2/d x 3 was the MTD, with dose-limiting hypotension at 98 mg/m2/d x 3. Based on chronic tolerability, 50 mg/m2/d x 3 is the recommended phase II dose without ADP. Antitumor effect was observed in certain patients with renal, prostate, and colon cancer, and non-Hodgkin's lymphoma. Concentrations of flavopiridol (200 to 400 nM) needed for cyclin-dependent kinase inhibition in preclinical models were achieved safely.

Treatment of refractory myositis: a randomized crossover study of two new cytotoxic regimens.

OBJECTIVE: To assess the clinical usefulness of 2 novel therapies for treatment-resistant myositis. METHODS: Thirty patients with refractory myositis, of whom 25 had an inadequate or no response to previous cytotoxic therapy, were randomized to begin either a combination of weekly oral methotrexate and daily azathioprine (MTX/AZA) or intravenous methotrexate with leucovorin rescue (I.V. MTX) every 2 weeks for 6 months. Crossover to the alternate therapy occurred according to defined rules; evaluations of muscle strength and functional abilities were performed at the beginning, and after 3 and 6 months, of each treatment. RESULTS: Of the 15 patients initially randomized to oral MTX/AZA, 8 improved with oral therapy and 1 improved with I.V. MTX during the crossover period. Of the 15 patients initially randomized to I.V. MTX therapy, 3 improved with the I.V. therapy and 4 with the oral combination during the crossover period. Although the study lacked the power to directly compare both treatments, intention-to-treat analysis showed a trend in favor of those patients who first received oral combination therapy (P = 0.025). There were 0.09 adverse events per patient-month with oral combination therapy and 0.16 per patient-month with I.V. therapy (P = 0.09). CONCLUSION: Combination oral MTX/AZA may benefit patients with treatment-resistant myositis, including those who previously had inadequate responses to either MTX or AZA alone. I.V. MTX with leucovorin rescue may also benefit some patients with refractory myositis.

Combinations of paclitaxel and vinblastine and their effects on tubulin polymerization and cellular cytotoxicity: characterization of a synergistic schedule.

Paclitaxel (PTX) and vinblastine (VBL) represent 2 classes of drugs that target tubulin but have separate binding properties and opposing mechanisms of action. To evaluate the potential use of these agents together in a chemotherapeutic regimen, we investigated their effects on the dynamics of tubulin polymerization and cellular cytotoxicity, when administered singly or in combination. In human epidermoid carcinoma KB cells and MCF-7 breast carcinoma cells, we observed a time- and dose-dependent effect on cytoskeletal dynamics for both PTX and VBL. Tubulin polymerization induced by PTX was stable for more than 24 hr. When PTX treatment was followed by VBL, a time- and dose-dependent reversal of tubulin polymerization was observed. In contrast, rapid tubulin polymerization occurred when VBL was followed by PTX. When both agents were added simultaneously, a diminution of PTX-induced tubulin polymerization was observed with increasing doses of VBL; a maximum reduction was achieved when equal concentrations were used. Examination of the tubulin pattern by immunofluorescence in MCF-7 breast cancer cells confirmed and extended our findings. Bundle formation followed treatment with PTX. Addition of increasing concentrations of VBL prevented bundling; however, the normal cytoskeletal architecture was not restored. Cytotoxicity studies carried out using the median dose effect principles and the combination index analysis showed synergism when VBL and PTX were administered sequentially and antagonism for simultaneous administration. Our results demonstrate changes in tubulin dynamics following drug treatment and provide a rationale for combined PTX/VBL therapy after careful evaluation of the schedule of administration.

An intra-abdominal small round cell neoplasm with features of primitive neuroectodermal and desmoplastic round cell tumor and a EWS/FLI-1 fusion transcript.

We report an intra-abdominal round cell tumor in a young man which exhibited the light and electron microscopic appearance of a peripheral primitive neuroectodermal tumor (PNET), in addition to the clinical and topographic characteristics, desmoplasia and a complex immunophenotypic profile of the intra-abdominal desmoplastic round cell tumor (DSRCT). Reverse transcription polymerase chain reaction revealed a EWS/FLI-1 fusion transcript as in PNET/Ewing's sarcoma, instead of the EWS/WT1 transcript of DSRCT. The tumor was also strongly positive for the mic2 protein. This is a unique case of a hybrid tumor arising in the peritoneal cavity of a young male. The existence of such a hybrid tumor in this location suggests that DSRCT and PNET may be related and possibly share a common histogenesis.

Update on therapy for refractory dermatomyositis and polymyositis.

Evaluation of new therapies for the inflammatory myopathies is complicated by the heterogeneity of these syndromes as well as by the lack of internationally accepted definitions of disease categories and assessments of disease activity and chronicity. This review covers our opinion of therapies and emphasizes the need for an early rehabilitation evaluation for these patients. Oral corticosteroids are the first line of therapy for the inflammatory myopathies, but because of their side effects and the existence of a subset of patients in whom disease is controlled only with high-dose corticosteroids, we recommend considering the early use of a second-line immunomodulating agents or pulse intravenous methylprednisolone. A stepwise progression of therapies is suggested for patients who have increasing muscle weakness resulting from active disease.

Cutaneous ulcers due to Candida albicans in an immunocompromised patient--response to therapy with itraconazole.

The patient, a 68-year-old female, with chronic anaemia and a previous history of carcinoma of the breast treated surgically, had been receiving therapy with methyl prednisone, in addition to gold, for pemphigus vulgaris. She developed deep cutaneous ulcers on the lower leg. All had well-defined edges and were covered with purulent and serosanguinous exudates. On histopathology the ulcers were deeply infiltrated with yeasts and mycelium and Candida albicans was isolated on culture. There was no evidence of systemic candidosis. Complete healing was obtained using itraconazole in a dose of 200 mg daily for 45 days: treatment with prednisone was continued throughout. The response to antifungal therapy alone suggests that Candida was largely, if not wholly responsible, for this unusual clinical condition.