Development of a novel UHPLC-MS/MS method for the determination of ochratoxin A in tea.

The mycotoxin Ochratoxin A (OTA) is responsible for producing many effects on human and animal health. In this work, the evaluation of the presence of OTA in tea beverage samples consisted of extraction and preconcentration through the solidification of a floating organic drop (DLLME-SFO) combined with an additional octadecyl silane clean-up step. The obtained extract was analyzed by UHPLC-MS/MS. Interferences from the matrix were effectively reduced and, consequently, recovery increased from 43.18% ± 4.1%-96.02% ± 2.54%. The validation assays were carried out by external calibration and spiked samples, with satisfactory recoveries. An adequate dynamic calibration range was obtained over a concentration interval between 0.5 and 70 µg mL-1 OTA. Capabilities of detection and quantification were 0.5 and 1.4 µg mL-1. The obtained Green Certificate was compared with other techniques to establish the greenness profile of the procedure. Quantification of ochratoxin A levels in tea samples was performed.

DLLME-SFO-GC-MS procedure for the determination of 10 organochlorine pesticides in water and remediation using magnetite nanoparticles.

There exists a high demand for fast, simple, and reliable methodologies for determining the presence of organochlorine pesticides (OCPs) on environmental samples. Moreover, the toxicity and accumulation of potential OCPs in several environments have led to the development of technologies that achieve their removal from contaminated waters. In this study, a novel method combining a dispersive liquid-liquid microextraction procedure based on the solidification of floating organic drop is developed and validated for the extraction, preconcentration, and determination of 10 OCPs: α-BHC, p,p'-DDE, δ-BHC, dieldrin, p,p'-DDT, endosulfan I, endosulfan sulfate, heptachlor, heptachlor epoxide (isomer B), and methoxychlor in water samples. The results show that the calibration curves were linear for all the studied compounds, and the coefficients of correlation higher than 0.99. The variation coefficient for precision and accuracy was lower than 10%, and the accuracy ranged from 93 to 105%. Low limit of detection and limit of quantification values ranging from 0.06-3.00 ng mL-1 and 0.20-10 ng mL-1 were obtained, respectively. The capability of the proposed method was confirmed using an analysis of the water samples before and after the degradation process; this was achieved by employing nanomaterials, while performing an analysis of 160 real samples that were sourced from a Brazilian river. A cobalt-doped magnetite was applied for the environmental remediation of the studied compounds, and it was verified that the novel material has the potential to be used in environmental remediation with a degradation efficiency exceeding 80% for the majority of the studied compounds.

Lesion of the commissural nucleus of the solitary tract/A2 noradrenergic neurons facilitates the activation of angiotensinergic mechanisms in response to hemorrhage.

In the present study, we investigated the effects of lesions of A2 neurons of the commissural nucleus of the solitary tract (cNTS) alone or combined with the blockade of angiotensinergic mechanisms on the recovery of arterial pressure (AP) to hemorrhage in conscious rats. Male Holtzman rats (280-320g) received an injection of anti-dopamine-beta-hydroxylase-saporin (12.6ng/60nl; cNTS/A2-lesion, n=28) or immunoglobulin G (IgG)-saporin (12.6ng/60nl, sham, n=24) into the cNTS and 15-21days later had a stainless steel cannula implanted in the lateral ventricle. After 6days, rats were submitted to hemorrhage (four blood withdrawals, 2ml/300g of body weight every 10min). Both cNTS/A2-lesioned and sham rats had similar hypotension to hemorrhage (-62±7 and -73±7mmHg, respectively), however cNTS/A2-lesioned rats rapidly recovered from hypotension (-5±3mmHg at 30min), whereas sham rats did not completely recover until the end of the recording (-20±3mmHg at 60min). Losartan (angiotensin type 1 receptor antagonist) injected intracerebroventricularly (100µg/1µl) or intravenously (i.v.) (10mg/kg of body weight) impaired the recovery of AP in cNTS/A2-lesioned rats (-24±6 and -35±7mmHg at 30min, respectively). In sham rats, only i.v. losartan affected the recovery of AP (-39±6mmHg at 60min). The results suggest that lesion of the A2 neurons in the cNTS facilitates the activation of the angiotensinergic pressor mechanisms in response to hemorrhage.

Facilitation of sodium intake by combining noradrenaline into the lateral parabrachial nucleus with prazosin peripherally.

Injections of noradrenaline into the lateral parabrachial nucleus (LPBN) increase arterial pressure and 1.8% NaCl intake and decrease water intake in rats treated with the diuretic furosemide (FURO) combined with a low dose of the angiotensin converting enzyme inhibitor captopril (CAP). In the present study, we investigated the influence of the pressor response elicited by noradrenaline injected into the LPBN on FURO+CAP-induced water and 1.8% NaCl intake. Male Holtzman rats with bilateral stainless steel guide-cannulas implanted into LPBN were used. Bilateral injections of noradrenaline (40 nmol/0.2 µl) into the LPBN increased FURO+CAP-induced 1.8% NaCl intake (12.2±3.5, vs., saline: 4.2±0.8 ml/180 min), reduced water intake and strongly increased arterial pressure (50±7, vs. saline: 1±1 mmHg). The blockade of the α1 adrenoceptors with the prazosin injected intraperitoneally abolished the pressor response and increased 1.8% NaCl and water intake in rats treated with FURO+CAP combined with noradrenaline injected into the LPBN. The deactivation of baro and perhaps volume receptors due to the cardiovascular effects of prazosin is a mechanism that may facilitate water and NaCl intake in rats treated with FURO+CAP combined with noradrenaline injected into the LPBN. Therefore, the activation of α2 adrenoceptors with noradrenaline injected into the LPBN, at least in dose tested, may not completely remove the inhibitory signals produced by the activation of the cardiovascular receptors, particularly the signals that result from the extra activation of these receptors with the increase of arterial pressure.

Analysis of brain nuclei accessible to ghrelin present in the cerebrospinal fluid.

Ghrelin is a stomach-derived peptide hormone that acts in the brain to regulate many important physiological functions. Ghrelin receptor, named the growth hormone secretagogue receptor (GHSR), is present in many brain areas with or without obvious direct access to ghrelin circulating in the bloodstream. Ghrelin is also present in the cerebrospinal fluid (CSF) but the brain targets of CSF ghrelin are unclear. Here, we studied which brain areas are accessible to ghrelin present in the CSF. For this purpose, we centrally injected mice with fluorescein-labeled ghrelin (F-ghrelin) peptide tracer and then systematically mapped the distribution of F-ghrelin signal through the brain. Our results indicated that centrally injected F-ghrelin labels neurons in most of the brain areas where GHSR is present. Also, we detected F-ghrelin uptake in the ependymal cells of both wild-type and GHSR-null mice. We conclude that CSF ghrelin is able to reach most of brain areas expressing GHSR. Also, we propose that the accessibility of CSF ghrelin to the brain parenchyma occurs through the ependymal cells in a GHSR-independent manner.

The Edinger-Westphal nucleus II: Hypothalamic afferents in the rat.

Numerous functions have been attributed to the Edinger-Westphal nucleus (EW), including those related to feeding behavior, pain control, alcohol consumption and the stress response. The EW is thought to consist of two parts: one controls accommodation, choroidal blood flow and pupillary constriction, primarily comprising cholinergic cells and projecting to the ciliary ganglion; and the other would be involved in the non-ocular functions mentioned above, comprising peptide-producing neurons and projecting to the brainstem, spinal cord and prosencephalic regions. Despite the fact that the EW is well known, its connections have yet to be described in detail. The aim of this work was to produce a map of the hypothalamic sources of afferents to the EW in the rat. We injected the retrograde tracer Fluoro-Gold into the EW, and using biotinylated dextran amine, injected into afferent sources as the anterograde control. We found retrogradely labeled cells in the following regions: subfornical organ, paraventricular hypothalamic nucleus, arcuate nucleus, lateral hypothalamic area, zona incerta, posterior hypothalamic nucleus, medial vestibular nucleus and cerebellar interpositus nucleus. After injecting BDA into the paraventricular hypothalamic nucleus, lateral hypothalamic area and posterior hypothalamic nucleus, we found anterogradely labeled fibers in close apposition to and potential synaptic contact with urocortin 1-immunoreactive cells in the EW. On the basis of our findings, we can suggest that the connections between the EW and the hypothalamic nuclei are involved in controlling stress responses and feeding behavior.