Inhibition of Aspergillus carbonarius growth and Ochratoxin A production using lactic acid bacteria cultivated in an optimized medium.

The Gram-positive bacteria lactic acid bacteria (LAB) are used in the food industry but are also known for inhibiting certain food spoilage microorganisms, especially fungi. Sources of nitrogen (N) for culture media are generally organic and expensive. Many attempts have been made to formulate economical culture media with alternative N sources obtained from agricultural and industrial byproducts. This study describes the design and optimization of an inexpensive culture medium for Lactiplantibacillus plantarum (formerly Lactobacillus plantarum) MZ809351 strain B31. The culture medium was optimized using statistical experimental designs to identify the factors with the most significant effects on biomass concentration to reduce the overall cost, aiming to obtain a biomass concentration similar to that obtained with the reference LAB culture medium (de Man, Rogosa and Sharpe; MRS). Sodium acetate and magnesium sulfate were the most significant factors (p < 0.005), and their contents were reduced by 22 % and 40 %, respectively, without affecting biomass concentration. Malt germ extract (MGE) was used as an alternative nitrogen source to replace meat extract (ME) and proteose peptone (PP). Through these experiments, the composition of a culture medium that is less expensive than MRS broth was defined, which produced a biomass concentration (3.8 g/L) similar to that obtained with MRS medium. The inhibitory effects of two LAB strains isolated from the Ivory Coast and Mexico on the growth and production of ochratoxin A (OTA) in an ochratoxigenic fungus was tested. The minimum cellular concentration of the LAB to prevent the development of Aspergillus carbonarius Ac 089 and the production of OTA was determined in a model assay in Petri dishes. The conditions to inhibit the germination of A. carbonarius Ac 089 and the production of OTA were found. Using the optimized medium and a ratio of 2 × 104 LAB/spore (1 × 108 CFU/mL) strain B7 (L. plantarum MZ809351) and 2 × 103 LAB/spore (1 × 107 CFU/mL) strain B31 (L. plantarum MN922335) completely inhibited the growth of the fungus. A ratio of 2 × 105 LAB/spore (1 × 109 CFU/mL) was required to inhibit OTA production with strains B7 and B31. This study indicates the potential of cultivating LAB in an optimized and inexpensive culture medium for use as a biological control agent against ochratoxigenic fungi in food.

Transcriptomes of the interaction between Fusarium verticillioides and a Streptomyces strain reveal the fungal defense strategy under the pressure of a potential biocontrol agent.

The actinobacteria Streptomyces sp. AV05 appears to be a potential biocontrol agent (BCA) against mycotoxigenic fungi. It was found to significantly inhibit F. verticillioides growth and mycotoxin production during their co-cultivation. F. verticillioides growth was durably affected while the decrease of the toxin production levels was reversible, suggesting different BCA actions. The study of both transcriptomes brought useful information on the microbial interaction. RNA-seq data indicated that the dual interaction modified genetic expression of both microorganisms as 18.5 % of the genes were differentially expressed for the fungus against 3.8 % for the actinobacteria. Fungal differentially expressed genes (DEGs) were equally up and down regulated while bacterial ones were mainly upregulated. We especially focused the analysis of DEGs on fungal defense reaction to bacterial attack. For example, if this potential BCA implements a strategy of antibiosis with the over expression of 'siderophore-interacting protein' linked to the production of bacteriocins, the fungus in a state of stress is able to adapt its metabolism by up-regulation of amidase. It could correspond to the induction of resistance gene clusters and suggest a detoxification process. Moreover fumonisins-related pathway appears underexpressed in the presence of Streptomyces that explain the reduction of fumonisin accumulation observed.

Risk assessment of exposure to mycotoxins (aflatoxins and fumonisins) through corn tortilla intake in Veracruz City (Mexico).

This study aimed to assess aflatoxin and fumonisin intake through corn tortilla consumption in Veracruz city. Between October 2013 and February 2015, a total of 120 corn tortilla samples (2 kg samples, 40 samples per year) were randomly collected. Aflatoxins and fumonisins were quantified by high performance liquid chromatography coupled with a fluorescence detector. A probability density function (PDF) was used for describing corn tortilla intake, body weight of the Veracruz city population, mycotoxin content of corn tortilla samples and estimated mycotoxin daily intake. The Monte Carlo method with 10,000 iterations was employed to assess the population exposure risk. The highest level of total aflatoxins (AFT) was 22.17 µg kg-1, and 526.6 µg kg-1 for fumonisins B1 plus B2, with 85% and 90% of contaminated samples respectively. Up to 69.7 % of the population was estimated to consume a higher aflatoxin dose than that recommended by the JECFA (1 ng kg-1 of body weight per day); it was found that the recommended dose was exceeded to a greater extent in the male population, due to higher consumption of corn. The risk of fumonisin intake was less than 5 % due to the low presence and levels of these toxins in corn tortillas. The results suggest that corn tortilla consumers are at dietary risk caused by AFT contamination; this information should be considered when taking action to protect public health.

Evidence for a role of oestrogen receptor-related receptor in the regulation of male sexual behaviour in the moth Agrotis ipsilon.

The oestrogen receptor-related receptors (ERRs) are orphan nuclear receptors that were originally identified on the basis of their close homology to the oestrogen receptors. The three mammalian ERR genes participate in the regulation of vital physiological processes including reproduction, development and metabolic homeostasis. Although unique ERRs have been found in insects, data on the function and regulation of these receptors remain sparse. In the present study, a 2095-bp full-length cDNA encoding an ERR, termed AiERR, was isolated from males of the moth Agrotis ipsilon and deposited in the GenBank database under the accession number KT944662. The predicted AiERR protein shared an overall identity of 47-82% with other known insect and mammalian ERR homologues. AiERR exhibited a broad tissue expression pattern with the detection of one transcript of approximately 2 kb in the primary olfactory centres, the antennal lobes (AL). In adult males, the amount of AiERR mRNA in the AL increased concomitantly with age and responses to the female-emitted sex pheromone. Moreover, AiERR knockdown induced an inhibition in the sex pheromone-orientated flight of male. Using A. ipsilon as a model, our study demonstrates that the insect ERR is critical for the performance of male sexual behaviour, probably by acting on central pheromone processing.

Antennal uridine diphosphate (UDP)-glycosyltransferases in a pest insect: diversity and putative function in odorant and xenobiotics clearance.

Uridine diphosphate UDP-glycosyltransferases (UGTs) are detoxification enzymes widely distributed within living organisms. They are involved in the biotransformation of various lipophilic endogenous compounds and xenobiotics, including odorants. Several UGTs have been reported in the olfactory organs of mammals and involved in olfactory processing and detoxification within the olfactory mucosa but, in insects, this enzyme family is still poorly studied. Despite recent transcriptomic analyses, the diversity of antennal UGTs in insects has not been investigated. To date, only three UGT cDNAs have been shown to be expressed in insect olfactory organs. In the present study, we report the identification of eleven putative UGTs expressed in the antennae of the model pest insect Spodoptera littoralis. Phylogenetic analysis revealed that these UGTs belong to five different families, highlighting their structural diversity. In addition, two genes, UGT40R3 and UGT46A6, were either specifically expressed or overexpressed in the antennae, suggesting specific roles in this sensory organ. Exposure of male moths to the sex pheromone and to a plant odorant differentially downregulated the transcription levels of these two genes, revealing for the first time the regulation of insect UGTs by odorant exposure. Moreover, the specific antennal gene UGT46A6 was upregulated by insecticide topical application on antennae, suggesting its role in the protection of the olfactory organ towards xenobiotics. This work highlights the structural and functional diversity of UGTs within this highly specialized tissue.

Partial laryngectomy as salvage surgery after radiotherapy: oncological and functional outcomes and impact on quality of life. A retrospective study of 20 cases.

UNLABELLED: The gold standard for the management of laryngeal squamous cell carcinoma in a previously irradiated patient is "salvage" total laryngectomy, but surgical management by partial laryngectomy can sometimes be proposed in selected patients. OBJECTIVES: This study was designed to review the functional and oncological outcomes of patients treated by open partial laryngectomy for recurrent squamous cell carcinoma after failure of radiotherapy or involving previously irradiated tissues and to define prognostic criteria for the selection of patients eligible for this treatment strategy. MATERIALS AND METHODS: In this retrospective study, 20 patients underwent partial laryngectomy between 2000 and 2011 for recurrence or second primary stage I or II laryngeal squamous cell carcinoma in an irradiated territory (11 vertical partial laryngectomies; 9 horizontal partial laryngectomies). RESULTS: The 3-year overall survival rate in patients with negative resection margins was 66%, with higher survival rates for tumours confined to the glottis, and the 2-year local control rate was 67%. Positive resection margins requiring total laryngectomy were observed in 20% of cases. The 3-year overall survival rate was 56% in these patients. Exclusive oral feeding was restored in 75% of patients after an average of 32 days. The tracheotomy tube was removed after an average of 18 days in 90% of patients. The disease-free functional larynx preservation rate was 45%. CONCLUSIONS: Salvage partial laryngectomy in irradiated tissues is an alternative treatment option to total laryngectomy in selected patients.

Prognostic factors for secondary recurrence of pleomorphic adenoma: a 20-year, retrospective study.

PURPOSE: To evaluate the results of treatment of parotid pleomorphic adenoma, and the risk factors for secondary recurrence. MATERIALS AND METHODS: Single-centre, retrospective study of 32 patients with pleomorphic adenoma recurrence managed between 1988 and 2008. RESULTS: The mean age at diagnosis of primary pleomorphic adenoma recurrence was 43.4 years. Twenty-eight per cent of patients had secondary recurrence; 32 per cent had undergone two or more surgical resections and external adjuvant radiotherapy. An age of less than 25 years was significantly associated with an earlier primary recurrence (p = 0.008). The most significant histopathological risk factor for secondary recurrence was the presence of a multifocal tumour (p = 0.019). Other histopathological criteria (i.e. cellularity and capsule rupture) were not significant. Radiotherapy was not associated with a decrease in recurrence. Nine per cent of patients progressed to malignancy. The main surgical complication was definitive facial palsy (14 per cent). CONCLUSION: Pleomorphic adenoma recurrence requires surgery, with greatly increased risk to the facial nerve. Resection with clear surgical margins is required, especially in young patients with multifocal tumours. Radiotherapy may delay second recurrence in cases of multifocal tumour.

Adenoid cystic carcinoma of the paranasal sinuses: retrospective series and review of the literature.

OBJECTIVES: Adenoid cystic carcinomas (ACC) are rare malignant tumours arising in the major and minor salivary glands. Involvement of the nasal cavity and paranasal sinuses is rare and poorly described. The purpose of this study was to define the clinical and prognostic criteria of ACC of the paranasal sinuses based on the review of a series of 25 cases. MATERIAL AND METHODS: Single-centre retrospective study of 25 cases of ACC of the paranasal sinuses managed between 1998 and 2011, evaluating epidemiological, clinical, diagnostic and prognostic criteria. Factors influencing survival (Kaplan-Meier/Log Rank test) and the patient's quality of life (EORTC QLQ-C30 questionnaire) were also analysed. RESULTS: Most patients (72%) had a locally-advanced tumour (stage T3 or T4) at diagnosis. Tumour sites, in decreasing order of frequency, were the maxillary sinus, nasal cavities and ethmoid sinus. The most common presenting complaints were maxillary pain or heaviness, unilateral blocked nose, and repeated epistaxis. When the tumour was resectable, treatment comprised a combination of surgery and adjuvant radiotherapy. The 5-year overall survival rate was 63% and the 5-year disease-free survival rate was 43%. The TNM stage at diagnosis (P=0.03), the histological subtype (P=0.023), the possibility of combined surgery and radiotherapy (P=0.03), and local control (P=0.05) were significant factors of improved 5-year overall survival. Positive surgical margins were associated with a trend towards poorer 5-year disease-free survival (ns). CONCLUSIONS: ACC are rare malignant tumours associated with a poor prognosis, characterized by a high recurrence rate. Recommended treatment is a combination of surgery and adjuvant radiotherapy whenever possible. Five-year survival varies as a function of TNM stage, histological subtype, treatment options and local control.

The DeStress for Success Program: effects of a stress education program on cortisol levels and depressive symptomatology in adolescents making the transition to high school.

Various studies have shown that increased activity of the hypothalamic-pituitary-adrenal (HPA) axis can predict the onset of adolescent depressive symptomatology. We have previously shown that adolescents making the transition to high school present a significant increase in cortisol levels, the main product of HPA axis activation. In the present study, we evaluated whether a school-based education program developed according to the current state of knowledge on stress in psychoneuroendocrinology decreases cortisol levels and/or depressive symptoms in adolescents making the transition to high school. Participants were 504 Year 7 high school students from two private schools in the Montreal area. Adolescents of one school were exposed to the DeStress for Success Program while adolescents from the other school served as controls. Salivary cortisol levels and depressive symptomatology were measured before, immediately after as well as 3 months after exposure to the program. Measures of negative mood were obtained at baseline in order to determine whether adolescents starting high school with specific negative moods were differentially responsive to the program. The results show that only adolescents starting high school with high levels of anger responded to the intervention with a significant decrease in cortisol levels. Moreover, we found that adolescents who took part in the intervention and showed decreasing cortisol levels following the intervention (responders) were 2.45 times less at risk to suffer from clinical and subclinical depressive states three months post-intervention in comparison to adolescents who showed increasing cortisol levels following the intervention (nonresponders). This study provides the first evidence that a school-based program on stress is effective at decreasing cortisol levels and depressive symptomatology in adolescents making the transition to high school and it helps explain which adolescents are sensitive to the program and what are some of the characteristics of these individuals.

Synchronous bilateral acinic cell carcinoma of the parotid: when a tumor reveals another one.

INTRODUCTION: Acinic cell carcinoma is a rare form (2-4%) of parotid tumor. It is, however, the most frequent form of bilateral parotid tumor and the second most frequent form in children. CASE REPORT: A 22-year-old man had several years' history of left parotid tumor, discovered to be acinic cell carcinoma. MRI found a synchronous contralateral tumor of the same histologic nature. DISCUSSION/CONCLUSION: This case highlights the need for rigorous examination of the contralateral parotid in case of parotid tumor and especially of acinic cell carcinoma.

Clinical features and outcome of sphenoid sinus aspergillosis: A retrospective series of 15 cases.

INTRODUCTION: Sphenoid sinusitis is uncommon, and aspergillus infections represent a minority of these cases. This study was designed to describe the characteristics of this disease and present a review of the literature. PATIENTS AND METHOD: Retrospective study from 2004 to 2010 based on 15 patients managed at Nantes University Hospital. Risk factors and history, symptoms, intranasal examination, imaging, histological and mycological results as well as analysis of the response to treatment and outcome were analysed. RESULTS AND DISCUSSIONS: Patients were aged between 14 to 78 years, almost 75% of patients were older than 50 and 73% of patients were women. The most common symptoms were headache [80%], nasal blockage or discharge [33%], and recurrent mild epistaxis [20%]. Predisposing factors were immunodepression in three patients, with no cases of diabetes. Two patients had a history of intranasal surgery and one had a history of facial trauma. Nasal endoscopy was normal in 40% of cases. CT was suggestive of sphenoid sinus aspergillosis in more than one half of cases [8/15] and demonstrated osteolysis in four patients. An incorrect preoperative diagnosis of mucocele was proposed in three patients. Histological examination demonstrated spore-forming structures in every case, but culture was positive in only four cases. Only two patients required antifungal therapy, including one patient with invasive aspergillosis. CONCLUSION: Chronic noninvasive sphenoid sinus aspergillosis appears to be a benign disease, essentially affecting women and patients over the age of 50 years. Symptoms are fairly non-specific. Imaging and histological examination are essential for management. The invasive form is a serious disease requiring rapid, multidisciplinary management.

[Parapharyngeal tumors: management and postoperative results]. / Tumeurs de l'espace parapharyngé: prise en charge et résultats postopératoires.

OBJECTIVES: Parapharyngeal space tumours are rare and mostly benign, and their access is difficult. The aim of the study was to define the optimal treatment of these tumours. MATERIAL AND METHODS: Clinical retrospective study, with histological, treatment, prognosis analysis of the parapharyngeal space tumours occurred between 1994 and 2012 and the surgical approach considerations. RESULTS: 35 patients were included with a male sex ratio predominance (51.43%, p = ns). The mean age of diagnosis was 49 years. The majority of tumours were benign (75.76%, p < 0.001). 54.5% originated from salivary gland with a majority of pleomorphic adenomas (42.42%). The second most frequent aetiology found was schwannoma (21.21%). 2 patients refused the treatment. Accidentally discovered were frequent (28.57%). In the symptomatic forms, dysphagia dominated (25.71%). Oropharyngeal (20%) and/or cervical (17.14%) mass were rare. A CT scan and a MR imaging were performed in 82.86% and 71.43% of the cases. A fine needle aspiration cytology was achieved in 10 cases with a positive predictive value of 60%. The cervical submandibular trans digastric approach was the most accomplished (40%). Other approaches were also used: Trans oral (25.71%), parotidectomy (14.71%), combined trans oral and cervical or parotidectomy (14.71%) and the cervical approached associated to a mandibulotomy (5.71%). A complementary treatment was proposed in 85.7% of the cases of malignant tumours. Post-operative complications were dominated by Horner's syndrome (n = 3), a temporary facial palsy (n = 5), definitive (n = 1), a velar insufficiency (n = 1), persistent pain (n = 6). CONCLUSIONS: The presence of 25% of malignant tumours and 40% of pleomorphic adenomas require removal of parapharyngeal tumours, even if non symptomatic.

Surgical repair of labial defect.

The main cause of labial defect is carcinological, in 90% of cases involving the lower lip. Reconstruction for labial defect is a therapeutic challenge imposing two main requirements: to achieve good esthetic quality, and to conserve labial function. Steps of defect analysis is proposed. Very numerous techniques have been described that are not exhaustively explained in this surgical review. Thus, the authors suggest reliable, reproducible solutions that should enable the surgeon to deal with most of the labial defect situations encountered in practice, based on consensual indications. Node involvement and alternatives to surgery, such as X-ray therapy, also fall outside the field of the present article.

[Congenital cyst and fistula of the face and neck]. / Kystes et fistules congénitaux de la face et du cou.

The authors propose an update on cervicofacial congenital cysts and fistulas' symptomatology. Embryological data, epidemiology and clinical manifestations are described. A reminder of the therapeutic principles is proposed as well as the evolution of these congenital pathologies, which may or may not involve the branchial system. branchial.

A diversity of putative carboxylesterases are expressed in the antennae of the noctuid moth Spodoptera littoralis.

Recent studies have suggested that pheromone-degrading enzymes belonging to the carboxylesterase family could play a role in the dynamics of the olfactory response to acetate sex pheromones in insects. Bioinformatic analyses of a male antennal expressed sequence tag library allowed the identification of 19 putative esterase genes expressed in the antennae of the moth Spodoptera littoralis. Phylogenetic analysis revealed that these genes belong to different insect esterase clades, defined by their putative cellular localization and substrate preferences. Interestingly, two of the 19 genes appeared to be antennal specific, suggesting a specific role in olfactory processing. This high esterase diversity suggested that the antennae are the location for intense esterase-based metabolism, against potentially a large range of exogenous and endogenous molecules.

Effect of operating conditions on ochratoxin A extraction from roasted coffee.

Operating conditions affect ochratoxin A (OTA) extraction from roasted coffee. The OTA content found in the beverage can thus be greater than that found in the roasted coffee used to prepare it. Three extraction parameters were studied for roasted coffee: type of extraction solvent (alkaline, neutral, acid), temperature (ambient temperature/23 degrees C, 60 degrees C and 85 degrees C), and extraction time (5, 20, 30, 40, 50, 60 and 80 min). The alkaline solvent used in the method recommended by the European Union extracted OTA better, but a maximum content was obtained at 60 degrees C after 50 min. At least a 100% improvement in extraction was obtained when compared with the European Union usual quantification method that is carried out at ambient temperature. It turned out that the OTA extraction parameters for roasted coffee, as defined by that method, were not optimum and needed to be modified. These results were verified in double-extraction experiments showing that OTA is not completely extracted by this method. Confirmation was obtained by comparison of extraction methods on several commercial samples of roasted coffee.

A conserved acidic motif in the N-terminal domain of nitrate reductase is necessary for the inactivation of the enzyme in the dark by phosphorylation and 14-3-3 binding.

It has previously been shown that the N-terminal domain of tobacco (Nicotiana tabacum) nitrate reductase (NR) is involved in the inactivation of the enzyme by phosphorylation, which occurs in the dark (L. Nussaume, M. Vincentz, C. Meyer, J.P. Boutin, and M. Caboche [1995] Plant Cell 7: 611-621). The activity of a mutant NR protein lacking this N-terminal domain was no longer regulated by light-dark transitions. In this study smaller deletions were performed in the N-terminal domain of tobacco NR that removed protein motifs conserved among higher plant NRs. The resulting truncated NR-coding sequences were then fused to the cauliflower mosaic virus 35S RNA promoter and introduced in NR-deficient mutants of the closely related species Nicotiana plumbaginifolia. We found that the deletion of a conserved stretch of acidic residues led to an active NR protein that was more thermosensitive than the wild-type enzyme, but it was relatively insensitive to the inactivation by phosphorylation in the dark. Therefore, the removal of this acidic stretch seems to have the same effects on NR activation state as the deletion of the N-terminal domain. A hypothetical explanation for these observations is that a specific factor that impedes inactivation remains bound to the truncated enzyme. A synthetic peptide derived from this acidic protein motif was also found to be a good substrate for casein kinase II.

Randomly amplified polymorphic DNAs assess recombination following an induced parasexual cycle in Penicillium roqueforti.

Random amplified polymorphic DNAs (RAPDs) were used as a genetic marker system to characterize recombinant strains following the parasexual cycle of Penicillium roqueforti. After protoplast fusion and haploidization of diploid hybrids, segregants characterized by a reassortment of the parental genetic markers displayed specific RAPD fingerprints. The appearance or the loss of RAPD fragments demonstrate that these markers provide an efficient method to analyze recombination and to characterize somatic hybrids.