Beneficial effects of (RS)-glucoraphanin on the tight junction dysfunction in a mouse model of restraint stress.

AIMS: The purpose of this work is to evaluate the effects of (RS)-glucoraphanin, a glucosinolate present in Brassicaceae, notably in Tuscan black kale, and bioactivated with myrosinase enzyme (bioactive RS-GRA) (10mg/kg intraperitoneally), and to assess its capacity to prevent the dysfunction of the blood-brain barrier (BBB), a fundamental structure for brain homeostasis, in a mouse model of restraint stress. MAIN METHODS: CD1 mice were subjected to restraint stress by blocking the body with a tape on a table for 150 min at the four extremities. After the sacrifice of the animals, stomachs and brains were collected to perform histological evaluation, Evan's blue dye, immunohistochemistry and western blotting analysis, to evaluate whether immobilization stress leads to alterations of tight junction (TJ) components, such as claudin-1, claudin-3 and ZO-1. KEY FINDINGS: Immobilization causes considerable damage to BBB as shown by detection of Evan's blue dye, indicating a high level of extravasation due to stress. BBB alterations were accompanied by an enhancement of GFAP expression, IkB-alpha degradation followed by increased NF-kBp65 nuclear translocation, as well as caspase 3 overexpression. Conversely, our results revealed that bioactive RS-GRA treatment significantly counteracts the changes in all these parameters and preserves TJ integrity reducing the production of pro-inflammatory cytokines, such as TNF-α and IL-1ß, and increasing the production of IL-10, an anti-inflammatory cytokine. Additionally, bioactive RS-GRA shows antioxidant properties modulating iNOS and nitrotyrosine expression. SIGNIFICANCE: Our results clearly show that bioactive RS-GRA could represent a possible treatment during pharmacological therapy of stress.

Evaluation of olfactory dysfunction in neurodegenerative diseases.

It is known that the olfactory dysfunction is involved in various neurological diseases, such as Parkinson's disease, Alzheimer's disease, multiple sclerosis, Huntington's disease and motor neuron disease. In particular, the ability to identify and discriminate the odors, as well as the odor threshold, can be altered in these disorders. These changes often occur as early manifestation of the pathology and they are not always diagnosed on time. The aim of this review is to summarize the major neurological diseases which are preceded or accompanied by olfactory dysfunction. In addition, new instrumental approaches, such as psychophysical testing, olfactory event-related potentials (OERPs) and functional magnetic resonance imaging (fMRI) measurements, supported by olfactometer for the stimuli delivery, and their combination in evaluation of olfactory function will be discussed. In particular, OERPs and fMRI might to be good candidates to become useful additional tools in clinical protocols for early diagnosis of neurological diseases.

Predictive biomarkers of recovery in traumatic brain injury.

Recent advances in medicine, intensive care and diagnostic imaging modalities have led to a pronounced reduction in deaths and disability resulting from traumatic brain injury. However, there are not sufficient findings to evaluate and quantify the severity of the initial and secondary processes destructive and therefore there are not effective therapeutic measures to effectively predict the outcome. For this reason, in recent decades, researchers and clinicians have focused on specific markers of cellular brain injury to improve the diagnosis and the evaluation of outcome. Many proteins synthesized in the astroglia cells or in the neurons, such as neuron-specific enolase, S100 calcium binding protein B, myelin basic protein, creatine kinase brain isoenzyme, glial fibrillary acidic protein, plasma desoxyribonucleic acid, brain-derived neurotrophic factor, and ubiquitin carboxy-terminal hydrolase-L1, have been proposed as potential markers for cell damage in central nervous system. Usually, the levels of these proteins increase following brain injury and are found in increasing concentrations in the cerebrospinal fluid depending on the injury magnitude, and can also be found in blood stream because of a compromised blood-brain barrier. In this review, we examine the various factors that must be taken into account in the search for a reliable non-invasive biomarkers in traumatic brain injury and their role in the diagnosis and outcome evaluation.

A fluorescence-based sequence-specific primer PCR for the screening of HLA-B(*)57:01.

Abacavir (ABC) is an antiretroviral drug highly effective in the treatment of HIV, but its intake can cause severe hypersensitivity reaction (HSR). A strong association between HLA-B(*)57:01 and ABC HSRs was reported by several studies, which demonstrated that HLA-B(*)57:01 screening had a 100% negative predictive value and that it could accurately identify patients at high risk of ABC HSRs. We propose a new sequence-specific primer PCR assay based on fluorescence detection through CE which is highly sensitive, allowing the use of non-infective sources of DNA such as saliva and buccal swabs, in addition to blood and reproducible, allowing automation of the analytical process. The results of our study were first compared with a standard sequence-specific primer PCR technique and reported a concordance of 100%, and then a blind external validation further confirmed the accuracy of our method.

A multiplex molecular assay for the detection of uniparental disomy for human chromosome 7.

Uniparental disomy (UPD) describes the inheritance of both homologues of a pair of chromosomes from only one parent. This condition is often caused by nondisjunction events during meiosis. UPD has been reported as a rare cause of the autosomal recessive disorder and aberrant expression of imprinted genes that are expressed from only one parental allele, either maternal or paternal. Maternal and/or paternal UPD for chromosome 7 is the most frequently observed UPD after UPD15. Here we developed and validated, for the first time, an effective, CE-based method for a rapid and economic detection based on two-fluorescent STR multiplexes.