Inorganic elements in the fat bodies of Diatraea saccharalis (Lepidoptera: Crambidae) larvae parasitized by Cotesia flavipes (Hymenoptera: Braconidae).

Koinobiont parasitoids use several strategies to regulate the host's physiological processes during parasitism. Although many aspects of host-parasitoid interactions have been explored, studies that attempted to assess the effects of parasitism on the availability of inorganic elements in the host are virtually nonexistent. Therefore, we aimed to evaluate the effects of parasitism on the concentrations of inorganic elements in the fat bodies of larvae of Diatraea saccharalis (Lepidoptera: Crambidae) during the development of the parasitoid Cotesia flavipes (Hymenoptera: Braconidae), by using total reflection X-ray fluorescence (TXRF). TXRF analysis allowed comparisons of the changes in the availability of the elements P, S, K, Ca, Cr, Fe, Ni, Cu, and Zn in the fat body tissues of D. saccharalis larvae parasitized by C. flavipes. Overall, the concentration of inorganic elements was higher early in parasitoid development (1 and 3days after parasitism) compared to non-parasitized larvae, but much lower towards the end of parasitoid development (7 and 9days after parasitism). Ca, K, and S were reduced after the fifth day of parasitism, which affected the total abundance of inorganic elements observed in the fat bodies of the parasitized hosts. The regulatory mechanisms or pathological effects related to the observed variation of the host inorganic elements induced by the parasitoid remain unknown, but there might be a strategy to make these elements available to the parasitoid larvae at the end of their development, when higher metabolic activity of the host fat body is required to sustain parasitoid growth. The observed variation of the host's inorganic elements could also be related to the known effects of parasitism on the host's immune response.

Mitotic crossing-over induced by two commercial herbicides in diploid strains of the fungus Aspergillus nidulans.

Some herbicides are suspected of promoting teratogenic, carcinogenic and mutagenic events. Detection of induced mitotic crossing-over has proven to be an indirect way of testing the carcinogenic properties of suspicious substances, because mitotic crossing-over is involved in the multistep process of carcinogenesis. We examined mitotic crossing-over induced by two commercial herbicides (diuron and trifluralin) in diploid strains of Aspergillus nidulans based on the homozygotization index. Low doses (2.5 microg/mL) of diuron were sufficient to increase the mean homozygotization index in 2.1 and 11.3 times for UT448//UT196 and Dp II-I//UT196, respectively, whereas the same dose of trifluralin increased this mean only 1.2 (UT448//UT196) and 3.5 (Dp II-I//UT196) times, respectively. The lower homozygotization index value found for trifluralin could be due to its interference with mitotic crossing-over in eukaryotic cells. We concluded that the diploid Dp II-I//UT196 of A. nidulans is more sensitive to organic compounds than UT448//UT196; these compounds cause recombinational events at a greater frequency in the latter diploid. This system holds promise as an initial test for carcinogenicity of organic compounds, including herbicides.

Aspergillus nidulans as a biological system to detect the genotoxic effects of mercury fumes on eukaryotes.

Mercury (Hg) pollution is one of the most serious environmental problems. Due to public concern prompted by the symptoms displayed by people who consumed contaminated fish in Minamata, Japan in 1956, Hg pollution has since been kept under constant surveillance. However, despite considerable accumulation of knowledge on the noxious effects of ingested or inhaled Hg, especially for humans, there is virtually nothing known about the genotoxic effects of Hg. Because increased mitotic crossing over is assumed to be the first step leading to carcinogenesis, we used a sensitive short-term test (homozygotization index) to look for DNA alterations induced by Hg fumes. In one Aspergillus nidulans diploid strain (UT448//UT184), the effects of the Hg fumes appeared scattered all over the DNA, causing 3.05 times more recombination frequencies than the mean for other strains. Another diploid (Dp II-I//UT184) was little affected by Hg. This led us to hypothesize that a genetic factor present in the UT184 master strain genome, close to the nicB8 genetic marker, is responsible for this behavior. These findings corroborate our previous findings that the homozygotization index can be used as a bioassay for rapid and efficient assessment of ecotoxicological hazards.

Streptomyces sp. ASBV-1 reduces aflatoxin accumulation by Aspergillus parasiticus in peanut grains.

AIMS: To evaluate the ability of Streptomyces sp. (strain ASBV-1) to restrict aflatoxin accumulation in peanut grains. METHODS AND RESULTS: In the control of many phytopathogenic fungi the Streptomyces sp. ASBV-1 strain showed promise. An inhibitory test using this strain and A. parasiticus was conducted in peanut grains to evaluate the effects of this interaction on spore viability and aflatoxin accumulation. In some treatments the Streptomyces sp ASBV-1 strain reduced the viability of A. parasiticus spores by c. 85%, and inhibited aflatoxin accumulation in peanut grains. The values of these reductions ranged from 63 to 98% and from 67% to 96% for aflatoxins B(1) and G(1), respectively. CONCLUSIONS: It was demonstrated that Streptomyces sp. ASBV-1 is able to colonize peanut grains and thus inhibit the spore viability of A. parasiticus, as well as reducing aflatoxin production. SIGNIFICANCE AND IMPACT OF THE STUDY: The positive finding for aflatoxin accumulation reduction in peanut grains seems promising and suggests a wider use of this actinobacteria in biological control programmes.

Induction of mitotic crossing-over in diploid strains of Aspergillus nidulans using low-dose X-rays.

As a contribution towards detecting the genetic effects of low doses of genotoxic physical agents, this paper deals with the consequences of low-dose X-rays in the Aspergillus nidulans genome. The irradiation doses studied were those commonly used in dental clinics (1-5 cGy). Even very low doses promoted increased mitotic crossing-over frequencies in diploid strains heterozygous for several genetic markers including the ones involved in DNA repair and recombination mechanisms. Genetic markers of several heterozygous strains were individually analyzed disclosing that some markers were especially sensitive to the treatments. These markers should be chosen as bio-indicators in the homozygotization index assay to better detect the recombinogenic/carcinogenic genomic effects of low-dose X-rays.

Induction of mitotic crossing-over in diploid strains of Aspergillus nidulansusing low-dose X-rays

As a contribution towards detecting the genetic effects of low doses of genotoxic physical agents, this paper deals with the consequences of low-dose X-rays in the Aspergillus nidulans genome. The irradiation doses studied were those commonly used in dental clinics (1-5 cGy). Even very low doses promoted increased mitotic crossing-over frequencies in diploid strains heterozygous for several genetic markers including the ones involved in DNA repair and recombination mechanisms. Genetic markers of several heterozygous strains were individually analyzed disclosing that some markers were especially sensitive to the treatments. These markers should be chosen as bio-indicators in the homozygotization index assay to better detect the recombinogenic/carcinogenic genomic effects of low-dose X-rays.

A glycoprotein with anti-inflammatory properties secreted by an Aspergillus nidulans modified strain

Total RNA from lipopolysaccharide (LPS)-stimulated rat macrophages used to treat protoplasts from an Aspergillus nidulans strain originated the RT2 regenerated strain, whose culture supernatant showed anti-inflammatory activity in Wistar rats. The protein fraction presenting such anti-inflammatory activity was purified and biochemically identified. The screening of the fraction responsible for such anti-inflammatory property was performed by evaluating the inhibition of carrageenan-induced paw edema in male Swiss mice. Biochemical analyses of the anti-inflammatory protein used chromatography, carbohydrates quantification of the protein sample, amino acids content analysis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Total sugar quantification revealed 32 percent glycosylation of the protein fraction. Amino acid analysis of such fraction showed a peculiar pattern presenting 29 percent valine. SDS-PAGE revealed that the protein sample is pure and its molecular weight is about 40kDa. Intravenous injection of the isolated substance into mice significantly inhibited carrageenan-induced paw edema. The isolated glycoprotein decreased carrageenan-induced paw edema in a prostaglandin-dependent phase, suggesting an inhibitory effect of the isolated glycoprotein on prostaglandin synthesis.

Clinical epidemiology of eating disorders: results from the Sesto Fiorentino study.

BACKGROUND: It is speculated that clinical samples do not fully reflect the characteristics of eating disorders (EDs) as they are in the general population, especially in their lowest range of severity. The present article reports the prevalence of EDs in a community sample aged >14 years, their clinical and psychopathological features, and their course and outcome on naturalistic grounds. METHODS: The Sesto Fiorentino Study is a three-phase community-based survey where 2,355 out of 2,500 people representative of the population aged >14 years living in Sesto Fiorentino were evaluated by their own general practitioner using the Mini International Neuropsychiatric Interview plus six additional questions. All those who had positive results plus a probability sample of the non-cases were re-interviewed by psychiatrists using the Florence Psychiatric Interview. The subjects who reported ED symptoms were subsequently administered the Eating Disorder Examination (12th edition). RESULTS: Overall, the lifetime prevalence of EDs was 1.21%. More precisely, 0.42% had anorexia nervosa, 0.32% bulimia nervosa, 0.32% binge eating disorder and 0.32% eating disorder not otherwise specified. All the subjects suffering from an ED fulfilled diagnostic criteria for at least another DSM-IV axis I psychiatric disorder. At the moment of the interview, conducted a few years (average 7 years) after the onset of the disorder, 50% had fully recovered from EDs, 26.9% were currently affected by an ED, 23.1% showed a persistent body image disturbance and/or the presence of compensatory behaviours. CONCLUSIONS: Community surveys conducted by clinicians may provide useful additional information on the psychopathological features, natural course and outcome of these disorders on naturalistic grounds.

The Eating Disorder Examination as a retrospective interview.

UNLABELLED: The purpose of this study was to determine the validity of the Eating Disorder Examination 12.0D (EDE) when administered retrospectively. METHODS: Twenty-five female patients suffering from an eating disorder [(10 with anorexia nervosa (AN), 10 with bulimia nervosa (BN), 5 with eating disorders not otherwise specified (EDNOS)] were investigated using the EDE at the time of the first referral to our outpatient ward (T1). Afterwards (mean 1.4 +/- 0.6 years later) each patient was administered again the EDE by the same assessor (T2). At this time the interviewer asked the patients to answer the questions referring to the symptoms and behaviours at the time of the first interview. RESULTS: Test-retest correlation factors were 0.7 or greater for all subscales of the EDE (p < 0.0001) and 0.5 or greater for every single item of the EDE (p < 0.001), except for EDE 1.5 (snack after dinner) and EDE 9A.6 (maximum time free from objective bulimic episodes in the last two months). DISCUSSION: Our results provide evidence that the EDE 12.0D is a reliable interview even when administered retrospectively, suggesting the use of this instrument for the retrospective assessment of eating disorders.

Evaluation of the genotoxicity induced by the fungicide fenarimol in mammalian and plant cells by use of the single-cell gel electrophoresis assay.

Fenarimol, a systemic pyrimidine carbinol fungicide, is considered to be not genotoxic or weakly genotoxic, although the available toxicological data are controversial and incomplete. Our results obtained in vitro with leukocytes of two different rodent species (rat and mouse) show that fenarimol affects DNA, as detected by the single-cell gel electrophoresis (SCGE, Comet) assay. This fungicide is able to induce DNA damage in a dose-related manner, with significant effectiveness at 36 nM, but without significant interspecies differences. Simultaneous exposure of rat leukocytes to fenarimol (36-290 nM) and a model genotoxic compound (50 microg/ml bleomycin) produced a supra-additive cytotoxic and genotoxic effect. This supports previous findings suggesting possible co-toxic, co-mutagenic, cancer-promoting and co-carcinogenic potential of fenarimol, and modification of the effects of other xenobiotics found to be influenced by this agrotoxic chemical, with consequent different toxicological events. The potential for DNA strand breaks to act as a biomarker of genetic toxicity in plants in vivo was also considered, in view of the fact that higher plants represent reliable sensors in an ecosystem. Significant DNA breakage was observed in the nuclei of Impatiens balsamina leaves after in vivo treatment with fenarimol (145 nM, 1h). More than 50% of the cells showed such DNA damage.

Eating behavior in obese patients with and without type 2 diabetes mellitus.

OBJECTIVE: Aim of this study was the assessment of the prevalence of eating disorders, and of eating disorder symptoms, in obese patients with type 2 diabetes, compared to non-diabetic subjects. DESIGN: Three samples of individuals were studied: a series of 156 (76 male, 80 female) overweight and obese type 2 diabetic patients, aged 30-65 y, with a body mass index (BMI)>28 kg/m(2) (DM); a series of 192 (20 male, 172 female) obese (BMI>30 kg/m(2)) non-diabetic patients aged 30-65 y seeking treatment for weight loss (OC); and a non-clinical sample of 48 (22 male, 26 female) obese (BMI>30 kg/m(2)) subjects aged 30-65 y selected from the lists of two general practices (OP). Eating behavior was assessed using the Eating Disorder Examination (EDE 12.0D). RESULTS: The prevalence of Binge Eating Disorder was lower than 5% in all the three samples. Median EDE scores in females were significantly higher in OC (3.0) and OP (3.4) than in DM (1.7), while diabetic patients showed higher scores on Restraint than both non-diabetic samples. Among diabetic patients, a significant correlation of EDE scores with HbA(1)c was observed. CONCLUSIONS: Type 2 diabetes is unlikely to induce relevant eating disturbances in obese patients, apart from an increase in restraint. Abnormalities of eating attitudes and behavior are associated with an impairment of metabolic control.

Identification of Aspergillus nidulans genes essential for the accumulation of sterigmatocystin.

The fungus Aspergillus nidulans (Emericella nidulans) was used as a genetic model for the identification of genes required for efficient accumulation of sterigmatocystin (ST). The required gene for sterigmatocystin expression was stc, which is an intermediate penultimate product in the aflatoxin biosynthetic pathway. Genetic analysis included studies of the sexual and parasexual cycles. The allelic segregation rates and recombination frequencies between linked and nonlinked genetic markers were determined by the crossing of the strains UT448 (stc) to UT196 (stc(+)) and UT448 (stc) to UT184 (stc). Low ST accumulation (4.0 ppm) in the UT196 strain and in 7.4% of the meiotic segregants allowed us to map the stc locus at chromosome I, 3.4% distant from riboA1. The diploid UT448 (stc)//UT184 (stc), prepared from nonproducing strains, was analyzed based on the parasexual cycle, and 28% of the haploid segregants accumulated the ST toxin. The results suggest that UT448 carries the stc mutant (or an inactivated) allele and that UT184, although carrying the stc(+) allele, is reactivated only by the R2(+) factor, which is located at chromosome VIII of UT448. In such a configuration, the diploid accumulates large amounts of sterigmatocystin (40 ppm). Another regulator factor (R1), located at the meth-w (II) chromosomic interval, was identified in the UT448 strain. At DNA level in chromosome I, the R1 product acts and blocks the stcZ(+) gene transcription. In a different genotypic configuration, the R1 product interacts with the R2 product (of chromosome VIII), allowing the stcZ(+) gene expression. Furthermore, the diploid UT448 (stc)//UT196 (stc(+)) accumulated the ST toxin at high level (40 ppm), indicating similar interaction between mentioned factors and the stc gene. Obtained data suggest that R1 (II) regulates the stcZ(+) transcription, by interacting with chromosome I (at the DNA level) and that R2 (VIII) controls R1 activity at the cytoplasm level. Based on these results, we propose a regulation model for the sterigmatocystin production.

Psychopathological and clinical features of outpatients with an eating disorder not otherwise specified.

In order to investigate similarities and differences between Eating Disorder Not Otherwise Specified (EDNOS) and Anorexia Nervosa (AN) and Bulimia Nervosa (BN), we studied a consecutive series of 189 female outpatients attending two Eating Disorder Units. The data were collected by means of interviews (Eating Disorder Examination, EDE 12.0D), the Structured Diagnostic Interview for DSM III-R, (SCID), and self-reported questionnaires (Beck Depression Inventory, BDI, and State and Trait Anxiety Inventory, STAI 1-2). The diagnosis of EDNOS was as frequent as that of AN and BN (43.8% versus 43.2%). There were no significant differences between EDNOS and AN/BN patients in terms of their general and specific psychopathological features, but significant differences were observed between bulimic-like and anorectic-like EDNOS patients, as well as between those with AN and BN. In conclusion, in our clinical setting, the patients with EDNOS and those with typical eating disorders have similar psychopathological features, thus suggesting that EDNOS patients should be further divided into two groups, anorectic-like (similar to AN) and bulimic-like (similar to BN) patients.

Cognitive-behavioural therapy for bulimia nervosa and binge eating disorder. A review.

Cognitive-behavioural therapy (CBT) programmes for bulimia nervosa (BN) have been considerably refined during the last 2 decades, and such a treatment is now extensively used. The present paper describes the treatment rationale and structure, and reviews the available evidence on its efficacy. Compared to any other psychological or pharmacological treatment for which controlled studies have been published, CBT is reported to be more effective (the majority of studies), or at least as effective. A CBT programme for binge eating disorder (BED) has been created by adapting that of BN, but it has been less extensively evaluated in field trials. Even here, however, no other treatment has proven to be of greater efficacy than CBT. Various methodological limitations reduce the possibility of generalizing these findings. Moreover, CBT was found to be completely satisfactory neither for BN nor for BED, with moderate effectiveness and some limits. However, at the present state of treatment, no other therapeutical procedure seems to be more effective, more specific or more promising. It is speculated therefore that CBT could be presently considered the first-choice remedy for these severely disabling disorders.

Eating attitudes and behavior throughout the menstrual cycle in obese women: a case-control study.

Premenstrual variations of eating behavior are reported in several studies, but their relationship with mood is unclear. Eating behavior and physical and psychological complaints during the menstrual cycle were studied in 107 obese patients and 93 matched controls using retrospective (Weekly Bulimic Test Edinburgh, W-BITE and Premenstrual Assessment Form, PAF) and prospective (Daily Rating Form, DR) questionnaires. Eating disorder symptoms increased in the premenstrual phase, as shown by the W-BITE scores both in patients (6.2 +/- 5.3 premenstrual week vs 4.9 +/- 4.4 postmenstrual week, p < 0.05) and in control subjects (4.9 +/- 4.1 premenstrual week vs 4.2 +/- 3.0 postmenstrual week, p < 0.05) and were correlated to premenstrual complaints in control subjects (r = 0.5; p < 0.05) but not in obese women (r = 0.2; p = NS). A close relationship between physical and psychological premenstrual disturbances was observed in obese patients only. Premenstrual variation of eating behavior could be the target of specific treatment.

Epidemiology of social phobia: a clinical approach.

The recent epidemiologic studies report extremely varied rates for social phobia (SP). One of the reasons for this may be the difficulty in diagnosing SP, the boundaries of which are uncertain. A community survey was carried out using doctors with experience in clinical psychiatry as interviewers, and a clinical diagnostic instrument. Two thousand three hundred and fifty-five people (out of the 2,500 randomly selected from the population) living in Sesto Fiorentino, a suburb of Florence, Italy, were interviewed by their own general practitioner, using the MINI plus six additional questions. Six hundred and ten of the 623 subjects that were found positive for any form of psychopathology at the screening interview, and 57 negative subjects, were re-interviewed by residents in psychiatry using the Florence Psychiatric Interview (FPI). The FPI is a validated composite instrument that has the format of a structured clinical research record. It was found that 6.58% of subjects showed social anxiety not attributable to other psychiatric or medical conditions during their life. Social or occupational impairments meeting DSM-IV diagnostic requirements for SP was detected in 76 subjects (lifetime prevalence = 3.27%). Correction for age raises the lifetime expected prevalence to 4%. Sex ratio was approximately (F:M) 2:1. The most common fear was speaking in public (89.4%), followed by entering a room occupied by others (63.1%) and meeting with strangers (47.3%). Eighty-six point nine percent of subjects with SP complained of more than one fear. The mean age of onset (when the subjects first fully met DSM-IV criteria for SP) was 28.8 years, but the first symptoms of SP usually occurred much earlier, with a mean age of onset at 15.5 years. Ninety-two percent of cases with SP also showed at least one other co-morbid psychiatric disorder during their life. Lifetime prevalence of avoidant personality disorder (APD) was 3.6%. Forty-two point nine percent of cases with SP also had APD, whereas 37.9% of cases with APD developed SP.

Screening for binge eating disorder in obese outpatients.

The prevalence of binge eating disorder (BED) in clinical samples of obese patients is controversial, and sensitive diagnostic protocols for use in routine clinical practice need to be further defined. Three hundred forty-four obese (body mass index [BMI] > or =30 kg/m2) patients were studied with the Structured Clinical Interview for DSM-III-R to investigate the lifetime prevalence of mental disorders. The current prevalence of BED was assessed using DSM-IV criteria. Eating attitudes and behavior were investigated with the Bulimic Investigation Test, Edinburgh (BITE) and the Binge Eating Scale (BES). The Beck Depression Inventory (BDI) and Spielberg's State-Trait Anxiety Inventory (STAI) were also applied. The prevalence of BED was 7.5%. Patients with BED had a higher BMI compared with obese patients without BED. Differences in the lifetime prevalence of mental disorders in patients with and without BED were not statistically significant. Using the BES as a screening instrument for BED with a threshold of 17, the sensitivity was 84.8%, specificity 74.6%, positive predictive value 26.2%, and negative predictive value 97.9%. Using the BITE with a threshold of at least 10, the sensitivity was 91%, specificity 51.4%, positive predictive value 71.8%, and negative predictive value 98.2%. The BITE can be a valid alternative to the BES as a screening method for BED in obese patients.

Ribonucleic acid treatment alters gene expression in diploid strains of Aspergillus nidulans.

Physical and chemical agents that promote DNA damage can induce high levels of mitotic crossing-over in eukaryotic diploid cells. Similarly, foreign DNA segments introduced by transformation processes, in the cell genome, can also induce mitotic crossing-over as an outcome of the reactions leading to chromosomic balance or due to the mechanisms aiming at the integration of the exogenous DNA. Zucchi et al. have described a system showing that RNA treatments are capable of inducing changes in the genome of haploid receptor strains of Aspergillus nidulans. To verify the genetic consequences of this process in diploid cells, conidia from two strains of this fungus were protoplastized, treated with homologous RNA and analyzed. Alterations in the gene expression and in the mitotic crossing-over frequencies between linked markers were detected. Among the main observed effects there was a generalized alteration in gene expression which was very likely caused by a reversible gene inactivation mechanism due to the methylation of cytosine residues. This was confirmed by treating the haploid segregants with the hypomethylating agent 5-azacytidine, that restored the original gene activity. The presence of a duplicated segment in the chromosome I of one of the treated diploids, interfered with the RNA general effects on its genome.

RNA-mediated transformation in Aspergillus nidulans recovers gene functions lost by deletion or by point mutations.

This work presents the results on two different approaches of RNA-mediated transformation in A. nidulans: a) the receptor strain was an argB2 (III) mutant deficient in arginine (OTCase deficient), and b) the receptor was an A. nidulans mutant defective in nitrate reductase synthesis due to a deletion in the niaD gene (VIII). The analyses of the arg+ and the nia+ retrotransformants allowed an insight on the fate and inheritance of the newly acquired characteristics. The occurrence and the study of Gene Inactivation Mechanism (RIP-like) inactivating the expression of extra copies of genes ectopically scattered over the receptor genome, was a byproduct of this research. Retrotransformants were also used as RNA-donor for a second turn of retrotransformation of the argB and niaD receptor strains. Genetic analyses of the new retrotransformants proved that the retrotransformation ability is kept by the re-extracted RNA when used in a second round of transformation process. This is the best genetic evidence that the newly acquired genetic characteristics were cDNA inserted, precisely transcribed and expressed. These are the first in vivo evidences of genetic information transference mediated by homologous RNA in lower eukaryotes.