1,419 research outputs found
Antiepileptogenic effects of trilostane in the kainic acid model of temporal lobe epilepsy
Objective: Epileptogenesis after status epilepticus (SE) has a faster onset in rats treated to reduce brain levels of the anticonvulsant neurosteroid allopregnanolone with the 5α-reductase inhibitor finasteride; however, it still has to be evaluated whether treatments aimed at increasing allopregnanolone levels could result in the opposite effect of delaying epileptogenesis. This possibility could be tested using the peripherally active inhibitor of 3β-hydroxysteroid dehydrogenase/Δ5-4 isomerase trilostane, which has been shown repeatedly to increase allopregnanolone levels in the brain.
Methods: Trilostane (50 mg/kg) was administered subcutaneously once daily for
up to six consecutive days, starting 10 min after intraperitoneal administration of
kainic acid (15 mg/kg). Seizures were evaluated by video-electrocorticographic
recordings for 70 days maximum, and endogenous neurosteroid levels were
assessed by liquid chromatography–electrospray tandem mass spectrometry.
Immunohistochemical staining was performed to evaluate the presence of brain
lesions.
Results: Trilostane did not alter the latency of kainic acid-induced SE onset or its
overall duration. When compared to the vehicle-treated group, rats receiving six
daily trilostane injections presented a remarkable delay of the first spontaneous
electrocorticographic seizure and subsequent tonic–clonic spontaneous recurrent
seizures (SRSs). Conversely, rats treated with only the first trilostane injection
during SE did not differ from vehicle-treated rats in developing the SRSs.
Notably, trilostane did not modify neuronal cell densities or the overall damage
in the hippocampus. In comparison to the vehicle group, repeated administration
of trilostane significantly decreased the activated microglia morphology in
the subiculum. As expected, allopregnanolone and other neurosteroid levels were
remarkably increased in the hippocampus and neocortex of rats treated for 6 days
with trilostane, but pregnanolone was barely detectable. Neurosteroids returned
to basal levels after a week of trilostane washout..
Significance: Overall, these results suggest that trilostane led to a remarkable increase in allopregnanolone brain levels, which was associated with protracted effects on epileptogenesis
The first record in Italy of Trichogramma cordubense Vargas & Cabello 1985 (Hymenoptera: Trichogrammatidae) emerging from the eggs of Lobesia botrana (Denis & SchiffermĂĽller, 1775) (Lepidoptera: Tortricidae)
This study investigated the egg parasitoids of Lobesia botrana (Denis & SchiffermĂĽller, 1775) feeding on Daphne gnidium L. (Malvales, Thymelaeaceae) in the San Rossore-Migliarino-Massaciuccoli Nature Reserve (Tuscany, Italy). Four species of egg parasitoids of the genus Trichogramma spp. were obtained. The parasitization rate gradually increased over the season, reaching its maximum level in September 2015, with a percentage of parasitized eggs close to 55 %. Three of the species obtained were already known as L. botrana parasitoids, whereas the finding of Trichogramma cordubense Vargas & Cabello, 1985 represents the first recording in Italy, as well as the first report of this species among the natural enemies of L. botrana
Investigation on the microbiological hazards in an artisanal soft cheese produced in northern Italy and its production environment in different seasonal periods
The present study aimed at assessing the occurrence of microbiological hazards (Listeria monocytogenes, Staphylococcus aureus, Salmonella spp. and Escherichia coli O157) in an artisanal soft cheese produced in northern Italy. In the same product total bacterial count, lactic acid bacteria and Enterobacteriaceae were enumerated, and pH and water activity measured in two batches sampled in summer and winter. Samples of raw materials, environmental swabs from the production processes and cheese during 15 days of storage at 2 and 8°C as well as dynamic temperature of 2°C for 5 days and 8°C for 10 days were collected and tested. The load of total bacterial count was significantly higher in the winter batch in comparison to the summer one, with a significant increase at the end of the storage period also noticed for lactic acid bacteria. Statistical higher values of pH were registered in raw materials and end of storage in winter batch. S. aureus was con-firmed only in the winter batch within samples (n=4) of stored cheese. On plates used for E. coli O157 detection, colonies of Klebsiella pneumoniae and Klebsiella oxy-toca were isolated. The results suggest that the highest bacterial population in the winter batch was associated to a higher pH in stored cheese and a higher number of bio-logical hazards identified. Their isolation started in the maturation room suggesting this step as relevant for possible cheese contamination
Selective labeling: identifying representative sub-volumes for interactive segmentation
Automatic segmentation of challenging biomedical volumes with multiple objects is still an open research field. Automatic approaches usually require a large amount of training data to be able to model the complex and often noisy appearance and structure of biological organelles and their boundaries. However, due to the variety of different biological specimens and the large volume sizes of the datasets, training data is costly to produce, error prone and sparsely available. Here, we propose a novel Selective Labeling algorithm to overcome these challenges; an unsupervised sub-volume proposal method that identifies the most representative regions of a volume. This massively-reduced subset of regions are then manually labeled and combined with an active learning procedure to fully segment the volume. Results on a publicly available EM dataset demonstrate the quality of our approach by achieving equivalent segmentation accuracy with only 5 % of the training data
The resistome of commensal Escherichia coli isolated from broiler carcasses “produced without the use of antibiotics” -
Several strategies have been in place in food animal production to reduce the unnecessary use of antimicrobial agents. Beyond the monitoring of their use, the evaluation of the effect of these strategies on the occurrence and types of antimicrobial resistance (AMR) associated genes is crucial to untangle the potential emergence and spread of AMR to humans through the food chain. In the present study, the occurrence of these genes was evaluated in commensal Escherichia coli isolated from broiler carcasses “produced without the use of antibiotics” in three antibiotic-free (AB-free) farms in Italy in 2019. Sequenced data were analyzed along with publicly available genomes of E. coli collected in Italy from the broiler food chain from previous years (2017 to 2018). The genetic relationships among all 93 genomes were assessed on de novo assemblies by in silico MLST and SNP calling. Moreover, the resistomes of all genomes were investigated. According to SNP calling, genomes were gathered in three clades. Clade A encompassed, among others, ST117, ST8070 and ST1011 genomes. ST10 belonged to clade B, whereas Clade C included ST58, ST297, ST1101 and ST23 among others. Regarding the occurrence of AMR genes, a statistically significant lower occurrence of these genes in the genomes of this study in comparison to the public genomes was observed considering the whole group of genes as well as genes specifically conferring resistance to aminoglycosides, β-lactams, phenicols, trimethoprim and lincosamides. Moreover, significant reductions were observed by comparing the whole group of AMR associated mutations, as well as those specifically for fluoroquinolones and fosfomycin resistance. Although the identification of 3° generation cephalosporin resistance associated genes in AB-free E. coli is a concern, this study provides a first indication of the impact of a more prudent use of antimicrobial agents on the occurrence of AMR genes in Italian broiler production chain. More studies are needed in next years on a higher number of genomes to confirm this preliminary observation
Identification and synthesis of new sex-specific components of olive fruit fly (Bactrocera oleae) female rectal gland, through original Negishi reactions on supported catalysts
In the present study, eleven new sex-specific components extracted from female rectal gland of olive fruit flies were synthesized and identified. The quantitative determination of those components by GC and GC/EI-MS, at different moments of the insect life span, highlighted the growing trend of their secretion. While for the synthesis of saturated esters, conventional transesterification methods could be adopted, for the synthesis of unsaturated components, a Negishi cross-coupling between organozinc halides and (Z)-1-bromo-1-alkenes was developed. To the extent of our knowledge, this reaction represents the first example of supported-catalyst promoted Negishi coupling, between an alkylzinc reagent and an alkenyl halide
Occurrence of foodborne pathogens in Italian soft artisanal cheeses displaying different intra- and inter-batch variability of physicochemical and microbiological parameters
Artisanal cheeses are produced in small-scale production plants, where the lack of full automation and control of environmental and processing parameters suggests a potential risk of microbial contamination. The aim of this study was to perform a longitudinal survey in an Italian artisanal factory producing a spreadable soft cheese with no rind to evaluate the inter- and intra-batch variability of physicochemical and microbial parameters on a total of 720 environmental and cheese samples. Specifically on cheese samples, the evaluation was additionally performed on physicochemical parameters. Cheese samples were additionally collected during 15 days of storage at constant temperatures of 2 and 8 degrees C, as well as a dynamic profile of 2 degrees C for 5 days and 8 degrees C for 10 days. Furthermore, Enterobacteriaceae isolates were identified at species level to have a better knowledge of the environmental and cheese microbiota potentially harboring human pathogens. High inter-batch variability was observed for lactic acid bacteria (LAB) and total bacteria count (TBC) in cheese at the end of production but not for pH and water activity. A temperature of 8 degrees C was associated with a significantly higher load of Enterobacteriaceae in cheeses belonging to batch 6 at the end of storage, and this temperature also corresponded with the highest increase in LAB and TBC loads over cheese shelf life. Results from generalized linear mixed models (GLMMs) indicated that drains in the warm room and the packaging area were associated with higher levels of TBC and Enterobacteriaceae in cheese. Regarding foodborne pathogens, no sample was positive for verotoxigenic Escherichia coli (VTEC) or Listeria monocytogenes, whereas six Staphylococcus aureus and one Salmonella pullorum isolates were collected in cheese samples during storage and processing, respectively. Regarding Enterobacteriaceae, 166 isolates were identified at species level from all batches, with most isolates belonging to Klebsiella oxytoca and pneumoniae, Enterobacter cloacae, Hafnia alvei, and Citrobacter freundii evidencing the need to focus on standardizing the microbial quality of cow milk and on hygienic procedures for cleaning and disinfection especially in warm and maturation rooms. Further studies should be performed to investigate the potential pathogenicity and antimicrobial resistance of the identified Enterobacteriaceae species in artisanal cheeses
Augmentation of endogenous neurosteroid synthesis alters experimental status epilepticus dynamics
Neurosteroids can modulate Îł-aminobutyric acid type A receptor-mediated inhibitory
currents. Recently, we discovered that the neurosteroids progesterone,
5α-dihydroprogesterone, allopregnanolone, and pregnanolone are reduced in the cerebrospinal
fluid of patients with status epilepticus (SE). However, it is undetermined
whether neurosteroids influence SE. For this reason, first we evaluated whether the
inhibitor of adrenocortical steroid production trilostane (50 mg/kg) could modify the
levels of neurosteroids in the hippocampus and neocortex, and we found a remarkable
increase in pregnenolone, progesterone, 5α-dihydroprogesterone, and allopregnanolone
levels using liquid chromatography tandem mass spectrometry. Second, we
characterized the dynamics of SE in the presence of the varied neurosteroidal milieu
by a single intraperitoneal kainic acid (KA; 15 mg/kg) injection in trilostane-treated
rats and their controls. Convulsions started in advance in the trilostane group, already
appearing 90 minutes after the KA injection. In contrast to controls, convulsions
prevalently developed as generalized seizures with loss of posture in the trilostane
group. However, this effect was transient, and convulsions waned 2 hours before the
control group. Moreover, electrocorticographic traces of convulsions were shorter
in trilostane-treated rats, especially at the 180-minute (P < .001) and 210-minute
(P < .01) time points. These findings indicate that endogenous neurosteroids remarkably
modulate SE dynamics
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