Highly specific quantification of ergotamine in urine, blood, and hair samples by liquid chromatography-tandem mass spectrometry

Ergotamine has been used for therapeutic purposes since the 1950s, usually to treat vascular headache. It is highly toxic and in large, repeated doses can produce all the symptoms of ergot poisoning. A selective and sensitive method, based on liquid chromatography-tandem mass spectrometry (LC-MS2), has been developed for quantifying ergotamine in biological fluids with use of a quick and easy sample preparation. Ergotamine and the internal standard, trideuterated lysergic acid diethylamide, were extracted from human urine, blood, and hair by means of liquid-liquid extraction at alkaline pH. Gradient elution on a cyanopropyl column was used for chromatographic separation. Positive ion electrospray ionization and tandem mass spectrometry determination by collision-induced dissociation were performed in an ion trap mass spectrometer. The method was validated and successfully applied to a case of iatrogenic ergotism resulting from the intake of ergotamine tartrate for treating headache. For the first time, ergotamine was identified and quantified in hair. The ergotamine concentrations measured were 320 pg/mL in blood, 100 pg/mL in urine, 24 pg/mg in proximal hair, and 15 pg/mg in distal hair.     

Facial fear processing and psychotic symptoms in schizophrenia: functional magnetic resonance imaging study

BACKGROUND: The recognition of negative facial affect is impaired in people with schizophrenia. The neural underpinnings of this deficit and its relationship to the symptoms of psychosis are still unclear. AIMS: To examine the association between positive and negative psychotic symptoms and activation within the amygdala and extrastriate visual regions of patients with schizophrenia during fearful and neutral facial expression processing. METHOD: Functional magnetic resonance imaging was used to measure neural responses to neutral and fearful facial expressions in 11 patients with schizophrenia and 9 healthy volunteers during an implicit emotional task. RESULTS: No association between amygdala activation and positive symptoms was found; the activation within the left superior temporal gyrus was negatively associated with the negative symptoms of the patients. CONCLUSIONS: Our results indicate an association between impaired extrastriate visual processing of facial fear and negative symptoms, which may underlie the previously reported difficulties of patients with negative symptoms in the recognition of facial fear.     

Gender-related neuronal and smooth muscle damage of guinea pig isolated urinary bladder from anoxia-glucopenia and reperfusion injury and its relationship to glycogen content

AIMS: To investigate the effects of anoxia and glucopenia (A-G) on both male and female guinea pig urinary bladder. METHODS: In whole bladders superfused with oxygenated Krebs' solution, intrinsic nerves underwent electrical field stimulation (EFS) and smooth muscle stimulated with carbachol, ATP, and high potassium. The effect of 1, 2, or 3 hr A-G on the contractile response and the ensuing recovery in Krebs' solution, was monitored. Glycogen content in male and female urinary bladders was also measured. RESULTS: Under different stimuli male urinary bladder proved to contract more efficiently than female organ. After 1 hr A-G the EFS response of male urinary bladder was virtually abolished and returned to 60% of control response in the recovery phase; in female bladder the EFS responses fully recovered during the reperfusion phase. Full recovery of the response to carbachol, ATP, and high potassium stimulations was observed in both genders. A-G had to be extended to 2 hr to cause smooth muscle impairment (higher in male than in female) and a neuronal impairment in female urinary bladders. When 2-deoxyglucose (2-DG), an inhibitor of glycolysis, was added during 1 hr A-G, both neuronal and smooth muscle damages were significantly enhanced in male, as well as, though to a lesser extent, in female bladder. A significantly higher glycogen content was observed in female as compared to male bladders, which was inversely related with the duration of exposure to A-G. CONCLUSIONS: The higher resistance of female urinary bladder to A-G/reperfusion, can be partly ascribed to the higher glycogen content.     

The clinical significance of body composition and anthropometric evaluation in athletes

Body composition and anthropometric assessment provide the sports physician with useful information on the health state of the athlete and with some necessary elements to plan specific training loads in the most appropriate way. In practice, the chemical composition of an athlete’s body (especially those who carry out 1–2 daily workouts) is always in a physiologic condition that we can define as “dynamic” (concentration of electrolytes, hydration state and relationship between intra-and extra-cellular water, stages of growth of muscle mass and/or reduction of fat mass, etc.), with the exception of few times of year, such as the short resting break before resuming training. As a consequence, a real “baseline” (or “stationary”) physiological state, allowing to detect the parameters of body composition under the same conditions several times during the year, is only rarely achieved. In this paper, we wanted to review the most interesting parameters and methods for the evaluation of athletes’ body composition, and underline their potential applications, possible advantages, theoretical and practical limitations.     

Extracellular processing and presentation of a 69-mer synthetic polypetide to MHC class I-restricted T cells.

The classical pathway for MHC class-I-restricted Ag presentation processes cytosolic Ag synthesized in or delivered into the cytosol for binding to MHC class I molecules in the ER. Alternatively, Ag may be processed and bind class I molecules in endocytic compartments or at the cell surface after regurgitation of processed peptides. We show that a 69-mer synthetic polypeptide that carries the optimal 9-mer Kd-restricted epitope from the Plasmodium berghei circumsporozoite protein, PbCS 245-253, is presented to CD8+ T cells after a short incubation (1-2 h) with target cells. The presentation kinetics correlate with the length of the peptides when shorter peptide analogues are used. This presentation is independent of the transporters associated with antigen processing and presentation (TAP), does not require newly synthesized proteins and does not proceed via regurgitation of intracellularly processed peptides. In contrast, it is substantially decreased in the absence of beta2 microglobulin or serum. Taken together, these data suggest that serum components, such as proteases and beta2 microglobulin, allow the processing and loading of exogenous polypeptides onto empty cell surface class I molecules for presentation to CTL.     

Comparative radiographic evaluation of the alveolar bone crest after orthodontic treatment.

This study evaluated and compared the heights of the alveolar bone crests (AC) among orthodontic patients treated with either the simplified standard edgewise technique (group 1, n = 30), the edgewise straight-wire system (group 2, n = 30), or bioefficient therapy (group 3, n = 26). These 3 groups were compared with an untreated control group (group 4, n = 30). A comparison by sex of AC height was also conducted. The first premolars were extracted in every treated patient, and measurements were performed on bitewing radiographs taken after a mean posttreatment period of 2.17 years. The distances from the AC to the cementoenamel junction (CEJ) on the mesial and distal surfaces of the first molars and second premolars and on the distal surface of the canines were measured; the larger the distance, the greater the alveolar bone loss. The data were analyzed by 1-way analysis of variance and the Newman-Keuls test (P <.05) for comparison among the groups. Sex differences of the AC height were evaluated with the t test. All treated groups had larger, statistically significant CEJ-AC distances than the untreated group, primarily at the extraction areas. There were no consistent statistically significant differences in the areas among the treated groups. Mean distances of the CEJ-AC in boys were larger than or similar to those in girls. The patients in the treated groups showed a greater number of proximal surfaces with statistically significant differences between sexes, compared with the control subjects.     

Mutant torsinA, which causes early-onset primary torsion dystonia, is redistributed to membranous structures enriched in vesicular monoamine transporter in cultured human SH-SY5Y cells.

A single GAG deletion in the DYT1 gene causes primary early-onset, generalized torsion dystonia. The DYT1 protein product, torsinA, belongs to the AAA+ family of proteins. When overexpressed, wild-type torsinA localizes mainly to the endoplasmic reticulum, whereas the mutant forms inclusions of unclear biogenetic origin. In this study, overexpressed wild-type torsinA in human neuroblastoma (SH-SY5Y) cell lines was distributed throughout the cell body and colocalized with a marker for the endoplasmic reticulum, confirming it is an endoplasmic reticulum protein. However, mutant torsinA showed perinuclear staining and formed distinct globular inclusions, which did not colocalize with endoplasmic reticulum markers. Immunoelectron microscopy of the mutant torsinA inclusions revealed membrane whorls staining for torsinA, as well as labeling of lamellae, isolated bilayers, and perinuclear membranes. This finding shows that mutant torsinA redistributes to specific membranous structures, which may represent different stages of maturation of the intracellular inclusions. The mutant torsinA-containing bodies were immunoreactive for vesicular monoamine transporter 2 (VMAT2). VMAT2 expression is important for the exocytosis of bioactive monoamines in neurons. Abnormal processing, transport, or entrapment of VMAT2 within the mutant torsinA membranous inclusions, therefore, may affect cellular dopamine release, providing a potential pathogenic mechanism for the DYT1-dependent dystonia.