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1.
Sensors (Basel) ; 20(1)2019 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-31878178

RESUMO

Assessing levels of neuropeptide Y (NPY) in the human body has many medical uses. Accordingly, we report the quantitative detection of NPY biomarkers applying guided-mode resonance (GMR) biosensor methodology. The label-free sensor operates in the near-infrared spectral region exhibiting distinctive resonance signatures. The interaction of NPY with bioselective molecules on the sensor surface causes spectral shifts that directly identify the binding event without additional processing. In the experiments described here, NPY antibodies are attached to the sensor surface to impart specificity during operation. For the low concentrations of NPY of interest, we apply a sandwich NPY assay in which the sensor-linked anti-NPY molecule binds with NPY that subsequently binds with anti-NPY to close the sandwich. The sandwich assay achieves a detection limit of ~0.1 pM NPY. The photonic sensor methodology applied here enables expeditious high-throughput data acquisition with high sensitivity and specificity. The entire bioreaction is recorded as a function of time, in contrast to label-based methods with single-point detection. The convenient methodology and results reported are significant, as the NPY detection range of 0.1-10 pM demonstrated is useful in important medical circumstances.


Assuntos
Técnicas Biossensoriais/métodos , Neuropeptídeo Y/análise , Anticorpos Imobilizados/química , Anticorpos Imobilizados/imunologia , Biomarcadores/análise , Humanos , Imunoensaio , Neuropeptídeo Y/imunologia , Polímeros/química
2.
PLoS One ; 14(8): e0220749, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31415596

RESUMO

A method for assessing the relative sensitivity of research metrics is proposed and illustrated by comparing 18 outcome measures from a published study of the cognitive, mood, and hormonal effects of four different levels of stress induced by intense military training. Research on the human response to stress often assesses multiple disparate dependent measures. Selecting the most sensitive is difficult as formal methods to compare varied dependent measures have not been developed. The method first converts the outcome measures into standard scores (z-scores) and then compares them using analysis of variance to determine whether there are differences in how they assess the impact of graded levels of exposure to stress. The analysis detected various significant interactions in several measures and suggests self-report mood questionnaires were more sensitive to the stressors present in the study than the cognitive or hormonal measures which were used. These findings support the effectiveness of the z-score based method as a useful procedure for objectively evaluating the differential sensitivity of various metrics. This method could be useful for research on other independent variables when use of multiple assessment strategies is appropriate. It could be used for evaluating studies yielding conflicting results, such as those detecting effects on one parameter but not others. In such instances, cross-metric inconsistencies may be due to differential sensitivity of measurement strategies rather than actual differences in the effects of the independent-variable on the domains under investigation.


Assuntos
Afeto/fisiologia , Atenção/fisiologia , Cognição/fisiologia , Hidrocortisona/análise , Estresse Psicológico/psicologia , Adulto , Benchmarking , Fator Neurotrófico Derivado do Encéfalo/análise , Feminino , Humanos , Masculino , Neuropeptídeo Y/análise , Psicometria , Tempo de Reação/fisiologia , Saliva/química , Autorrelato , Inquéritos e Questionários , Testosterona/análise , Adulto Jovem
3.
Medicine (Baltimore) ; 98(13): e14982, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30921206

RESUMO

The purpose was to investigate changes in neuropeptide Y (NPY) protein and dipeptidyl peptidase IV (DPP-IV) activity in the plasma and saliva in normally cycling women and women after menopause. We recruited 7 cycling women and 7 postmenopausal women for a cross-sectional, prospective pilot study. Blood via venipuncture and saliva samples were taken at each point in the menstrual cycle (premenopausal) or once per week (postmenopausal) for 2 months. Blood and saliva were analyzed for estrogen, NPY using ELISA and DPP-IV activity using a fluorometric assay. Plasma ß-estradiol was an average of 96.45 ±â€Š57.04 pg/mL over 2 cycles in the premenopausal group and 1.72 ±â€Š0.35 pg/mL over 2 months in the postmenopausal group (P < .05). In the cycling group, there were no significant differences in saliva or plasma NPY or DPP-IV over the cycle. For the postmenopausal group, salivary NPY and DPP-IV did not change over 2 months. Plasma NPY was lowest in the middle 2 weeks (average: 0.52 ±â€Š0.10 ng/mL) compared to the first and fourth weeks (average of week 1 and 4: 0.60 ±â€Š0.14 ng/mL; P < .05). Plasma NPY in postmenopausal women was higher overall (0.56 ±â€Š0.13 ng/mL) compared to cycling women (0.30 ±â€Š0.11 ng/mL; P < .05). Plasma DPP-IV activity was unchanged by time in the postmenopausal group. Saliva DPP-IV and saliva NPY in the cycling group had a significant negative correlation (R = -0.95; P < .05). We found that saliva measures of NPY and DPP-IV activity appear to be poor estimates of plasma concentrations and activities, but a larger sample size is required to conform this. Differences in plasma NPY concentrations between the groups and the relationship between salivary NPY and DPP-IV suggests that there may be some unique differences between these groups.


Assuntos
Dipeptidil Peptidase 4/análise , Neuropeptídeo Y/análise , Pós-Menopausa/metabolismo , Pré-Menopausa/metabolismo , Saliva/química , Adulto , Estudos Transversais , Dipeptidil Peptidase 4/sangue , Feminino , Humanos , Pessoa de Meia-Idade , Neuropeptídeo Y/sangue , Projetos Piloto , Estudos Prospectivos
4.
ACS Appl Mater Interfaces ; 11(15): 13927-13934, 2019 Apr 17.
Artigo em Inglês | MEDLINE | ID: mdl-30884221

RESUMO

Transmission electron microscopy (TEM) is being pushed to new capabilities which enable studies on systems that were previously out of reach. Among recent innovations, TEM through liquid cells (LC-TEM) enables in operando observation of biological phenomena. This work applies LC-TEM to the study of biological components as they interact on an abiotic surface. Specifically, analytes or target molecules like neuropeptide Y (NPY) are observed in operando on functional graphene field-effect transistor (GFET) biosensors. Biological recognition elements (BREs) identified using biopanning with affinity to NPY are used to functionalize graphene to obtain selectivity. On working devices capable of achieving picomolar responsivity to neuropeptide Y, LC-TEM reveals translational motion, stochastic positional fluctuations due to constrained Brownian motion, and rotational dynamics of captured analyte. Coupling these observations with the electrical responses of the GFET biosensors in response to analyte capture and/or release will potentially enable new insights leading to more advanced and capable biosensor designs.


Assuntos
Técnicas Biossensoriais/métodos , Grafite/química , Neuropeptídeo Y/análise , Transistores Eletrônicos , Área Sob a Curva , Técnicas Biossensoriais/instrumentação , Microscopia Crioeletrônica , Humanos , Neuropeptídeo Y/química , Biblioteca de Peptídeos , Ligação Proteica , Curva ROC , Suor/metabolismo
5.
Anat Histol Embryol ; 47(6): 517-526, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30105873

RESUMO

The gastrointestinal (GI) tract is innervated by nerve processes derived from the intramural enteric neurons and neurons localized outside the digestive tract. This study analysed the neurochemical characterization of nerves in the wall of the porcine oesophagus using single immunofluorescence technique. Immunoreactivity to vesicular acetylcholine transporter (VAChT), neuropeptide Y (NPY), vasoactive intestinal polypeptide (VIP), somatostatin (SOM), galanin (GAL), neuronal isoform of nitric oxide synthase (nNOS), substance P (SP), leucine enkephalin (LENK), calcitonin gene-related peptide (CGRP) or dopamine beta-hydroxylase (DBH) was investigated in intramuscular and intramucosal nerves of the cervical, thoracic and abdominal oesophagus. The results indicate that all of the substances studied were present in the oesophageal nerves. The density of particular populations of fibres depended on the segment of the oesophagus. The most numerous were fibres immunoreactive to VIP in the longitudinal and circular muscle layers of the abdominal oesophagus: The number of these fibres amounted to 16.4 ± 0.8 and 18.1 ± 3.1, respectively. In turn, the least numerous were CGRP-positive fibres, which were present only in the circular muscle layer of the cervical oesophagus and mucosal layer of the abdominal oesophagus in the number of 0.3 ± 0. The obtained results show that nerves in the porcine oesophageal wall are very diverse in their neurochemical coding, and differences between particular parts of the oesophagus suggest that organization of the innervation clearly depends on the fragment of this organ.


Assuntos
Sistema Nervoso Entérico/química , Esôfago/inervação , Imunofluorescência/veterinária , Fibras Nervosas/química , Neuropeptídeos/análise , Animais , Peptídeo Relacionado com Gene de Calcitonina/análise , Dopamina beta-Hidroxilase/análise , Encefalina Leucina/análise , Feminino , Galanina/análise , Neuropeptídeo Y/análise , Óxido Nítrico Sintase Tipo I/análise , Somatostatina/análise , Substância P/análise , Suínos , Peptídeo Intestinal Vasoativo/análise , Proteínas Vesiculares de Transporte de Acetilcolina/análise
6.
J Neurosci ; 38(34): 7378-7391, 2018 08 22.
Artigo em Inglês | MEDLINE | ID: mdl-30037831

RESUMO

Development of cortical interneurons continues until the end of human pregnancy. Premature birth deprives the newborns from the supply of maternal estrogen and a secure intrauterine environment. Indeed, preterm infants suffer from neurobehavioral disorders. This can result from both preterm birth and associated postnatal complications, which might disrupt recruitment and maturation of cortical interneurons. We hypothesized that interneuron subtypes, including parvalbumin-positive (PV+), somatostatin-positive (SST+), calretinin-positive (CalR+), and neuropeptide Y-positive (NPY+) interneurons, were recruited in the upper and lower cortical layers in a distinct manner with advancing gestational age. In addition, preterm birth would disrupt the heterogeneity of cortical interneurons, which might be reversed by estrogen treatment. These hypotheses were tested by analyzing autopsy samples from premature infants and evaluating the effect of estrogen supplementation in prematurely delivered rabbits. The PV+ and CalR+ neurons were abundant, whereas SST+ and NPY+ neurons were few in cortical layers of preterm human infants. Premature birth of infants reduced the density of PV+ or GAD67+ neurons and increased SST+ interneurons in the upper cortical layers. Importantly, 17 ß-estradiol treatment in preterm rabbits increased the number of PV+ neurons in the upper cortical layers relative to controls at postnatal day 14 (P14) and P21 and transiently reduced SST population at P14. Moreover, protein and mRNA levels of Arx, a key regulator of cortical interneuron maturation and migration, were higher in estrogen-treated rabbits relative to controls. Therefore, deficits in PV+ and excess of SST+ neurons in premature newborns are ameliorated by estrogen replacement, which can be attributed to elevated Arx levels. Estrogen replacement might enhance neurodevelopmental outcomes in extremely preterm infants.SIGNIFICANCE STATEMENT Premature birth often leads to neurodevelopmental delays and behavioral disorders, which may be ascribed to disturbances in the development and maturation of cortical interneurons. Here, we show that preterm birth in humans is associated with reduced population of parvalbumin-positive (PV+) neurons and an excess of somatostatin-expressing interneurons in the cerebral cortex. More importantly, 17 ß-estradiol treatment increased the number of PV+ neurons in preterm-born rabbits, which appears to be mediated by an elevation in the expression of Arx transcription factor. Hence the present study highlights prematurity-induced reduction in PV+ neurons in human infants and reversal in their population by estrogen replacement in preterm rabbits. Because preterm birth drops plasma estrogen level 100-fold, estrogen replacement in extremely preterm infants might improve their developmental outcome and minimize neurobehavioral disorders.


Assuntos
Córtex Cerebral/patologia , Estradiol/farmacologia , Doenças do Prematuro/patologia , Interneurônios/efeitos dos fármacos , Animais , Animais Recém-Nascidos , Calbindina 2/análise , Contagem de Células , Feminino , Idade Gestacional , Glutamato Descarboxilase/análise , Humanos , Recém-Nascido , Recém-Nascido Prematuro , Interneurônios/química , Interneurônios/classificação , Interneurônios/fisiologia , Masculino , Proteínas do Tecido Nervoso/análise , Proteínas do Tecido Nervoso/biossíntese , Proteínas do Tecido Nervoso/genética , Neuropeptídeo Y/análise , Parvalbuminas/análise , Coelhos , Somatostatina/análise , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genética
7.
Domest Anim Endocrinol ; 65: 71-79, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30007131

RESUMO

Maternal nutrient restriction during gestation can exert long-term negative effects on offspring health and performance. Arginine supplementation may rescue some of the negative effects elicited by maternal nutrient restriction. We tested the hypothesis that maternal arginine supplementation during gestation would rescue deleterious effects of nutrient restriction on in vitro O2 consumption in the liver and jejunum and hypothalamic protein expression of proopiomelanocortin (POMC), neuropeptide Y (NPY), agouti-related peptide (AgRP), and neuronal nitric oxide synthase (nNOS), and the colocalization of nNOS and active phosphor-signal transducer and activator of transcription 3 (pSTAT3) in female offspring. Multiparous ewes were assigned to dietary treatment at 54 d of gestation: 100% of requirements (Con), 60% of control (Res), or Res plus rumen-protected arginine (Res-Arg; 180 mg/kg). At parturition, offspring were immediately removed from their dam and placed on a common diet. At 54 ± 4 d of age, female lambs (n = 6 per treatment) were weighed, the liver and jejunum were weighed, and samples were collected for in vitro measurement of O2 consumption. The hypothalamus was collected to determine protein expression of POMC, NPY, AgRP, and nNOS, and the colocalization of nNOS and pSTAT3 (n = 3, 4, and 4 for Con, Res, and Res-Arg, respectively). Hepatic consumption of O2 in vitro (mol/min/liver) was decreased (P = 0.04) in the Res and Res-Arg group compared with Con. Intensity of staining for NPY-containing fibers tended to decrease (P = 0.10) in Res and Res-Arg compared with Con. Number of POMC neuronal cells in the arcuate nucleus (ARC) of the hypothalamus decreased (P ≤ 0.03) in the Res group compared with Res-Arg. These observations demonstrate that maternal nutrient restriction decreases energy utilization in the liver and number of POMC cells in the ARC of offspring. Supplementation of arginine to the gestating ewe failed to influence hepatic use of energy in lambs from Res ewes. Numbers of POMC-containing cells were increased in the ARC in lambs from ewes restricted to 60% of nutritional requirements and supplemented with rumen-protected arginine, potentially influencing feeding behavior and hepatic energy metabolism.


Assuntos
Arginina/administração & dosagem , Privação de Alimentos/fisiologia , Hipotálamo/efeitos dos fármacos , Hipotálamo/fisiologia , Consumo de Oxigênio/efeitos dos fármacos , Ovinos/fisiologia , Proteína Relacionada com Agouti , Fenômenos Fisiológicos da Nutrição Animal , Animais , Dieta/veterinária , Suplementos Nutricionais , Feminino , Idade Gestacional , Hipotálamo/química , Imuno-Histoquímica , Jejuno/efeitos dos fármacos , Jejuno/metabolismo , Fígado/efeitos dos fármacos , Fígado/metabolismo , Fenômenos Fisiológicos da Nutrição Materna , Neuropeptídeo Y/análise , Óxido Nítrico Sintase Tipo I/análise , Necessidades Nutricionais , Gravidez , Pró-Opiomelanocortina/análise , Rúmen/metabolismo
8.
Mil Med ; 183(9-10): e349-e356, 2018 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-29547968

RESUMO

INTRODUCTION: Tactical Combat Casualty Care (TCCC) training imposes psychophysiological stress on medics. It is unclear whether these stress levels vary with the training modalities selected. It is also unclear how stress levels could have an impact on medical performance and skill uptake. MATERIALS AND METHODS: We conducted a pilot study to compare the effects of live tissue (LT) with a high-fidelity patient simulator (SIM) on the level of stress elicited, performance, and skill uptake during battlefield trauma training course in an operating room (OR) and in a simulated battlefield scenario (field). In the report, we studied the effects of training modalities and their changes on stress levels by measuring different biomarkers (salivary amylase, plasma catecholamines, and neuropeptide Y) at various time points during the trauma training course. RESULTS: We found that the training resulted in significant psychophysiological stress as indicated by elevated levels of various biomarkers relative to baseline immediately after both OR and field assessment (p < 0.05). Compared with pre-OR levels, the LT training in the OR resulted in significant increases in the plasma levels of epinephrine, norepinephrine, and neuropeptide (p = 0.013, 0.023, 0.004, respectively), whereas the SIM training in the OR resulted in significant increases in the plasma levels of norepinephrine and neuropeptide (p = 0.003 and 0.008). Compared with pre-field levels, we found significant increases in plasma epinephrine concentration in the SIM group (p = 0.016), plasma norepinephrine concentration in the LT group (p = 0.015), and plasma neuropeptide Y concentration in both LT (p = 0.006) and SIM groups (p = 0.029). No differences in the changes of biomarker levels were found between LT and SIM groups in the OR and field. Compared with pre-field levels, the testing on the same modality as that in the OR in the simulated battlefield resulted in significant increases in norepinephrine and neuropeptide levels (p = 0.013 and 0.015), whereas the testing on different modalities resulted in significant increases in amylase, epinephrine, and neuropeptide levels (p = 0.016, 0.05, 0.018, respectively). There was a significantly larger increase in plasma norepinephrine concentration (p = 0.031) and a trend toward a greater increase in the salivary amylase level (p = 0.052) when the field testing involved a different modality than the OR compared with when OR and field testing involved the same modality. Although most of the biomarkers returned to baseline levels after 24 h, plasma norepinephrine levels remained significantly higher regardless of whether field testing occurred on the same or different modality compared with OR (p = 0.040 and 0.002). CONCLUSION: TCCC training led to significant increase in psychophysiological stress, as indicated by elevated levels of various biomarkers. The training modalities did not result in any differences in stress levels, whereas the switch in training modalities appeared to elicit greater stress as evidenced by changes in specific biomarkers (amylase and norepinephrine). A comparative study with a larger sample size is warranted.


Assuntos
Biomarcadores/análise , Simulação de Paciente , Estresse Psicológico/complicações , Ensino/psicologia , Ferimentos e Lesões/complicações , Amilases/análise , Catecolaminas/análise , Humanos , Neuropeptídeo Y/análise , Salas Cirúrgicas/métodos , Projetos Piloto , Treinamento por Simulação/métodos , Estresse Psicológico/psicologia
9.
Brain Stimul ; 11(4): 797-805, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29519725

RESUMO

BACKGROUND: Repetitive transcranial magnetic stimulation (rTMS) is able to modify cortical excitability. Rat rTMS studies revealed a modulation of inhibitory systems, in particular that of the parvalbumin-expressing (PV+) interneurons, when using intermittent theta-burst stimulation (iTBS). OBJECTIVE: The potential disinhibitory action of iTBS raises the questions of how neocortical circuits stabilize excitatory-inhibitory balance within a physiological range. Neuropeptide Y (NPY) appears to be one candidate. METHODS: Analysis of cortical expression of PV, NPY and vesicular glutamate transporter type 1 (vGluT1) by immunohistochemical means at the level of cell counts, mean neuropil expression and single cell pre-/postsynaptic expression, with and without intraventricular NPY-injection. RESULTS: Our results show that iTBS not only reduced the number of neurons with high-PV expression in a dose-dependent fashion, but also increased the cortical expression of NPY, discussed to reduce glutamatergic transmission, and this was further associated with a reduced vGluT1 expression, an indicator of glutamateric presynaptic activity. Interneurons showing a low-PV expression exhibit less presynaptic vGluT1 expression compared to those with a high-PV expression. Intraventricular application of NPY prior to iTBS prevented the iTBS-induced reduction in the number of high-PV neurons, the reduction in tissue vGluT1 level and that presynaptic to high-PV cells. CONCLUSIONS: We conclude that NPY, possibly via a global but also slow homeostatic control of glutamatergic transmission, modulates the strength and direction of the iTBS effects, likely preventing pathological imbalance of excitatory and inhibitory cortical activity but still allowing enough disinhibition beneficial for plastic changes as during learning.


Assuntos
Córtex Cerebral/metabolismo , Excitabilidade Cortical/fisiologia , Homeostase/fisiologia , Neuropeptídeo Y/biossíntese , Estimulação Magnética Transcraniana/métodos , Potenciais de Ação/fisiologia , Animais , Córtex Cerebral/química , Interneurônios/química , Interneurônios/metabolismo , Aprendizagem/fisiologia , Masculino , Neurônios/química , Neurônios/metabolismo , Neuropeptídeo Y/análise , Parvalbuminas/análise , Parvalbuminas/metabolismo , Ratos , Ratos Sprague-Dawley , Proteína Vesicular 1 de Transporte de Glutamato/análise , Proteína Vesicular 1 de Transporte de Glutamato/metabolismo
10.
Elife ; 72018 02 27.
Artigo em Inglês | MEDLINE | ID: mdl-29485402

RESUMO

Down syndrome, the leading genetic cause of intellectual disability, results from an extra-copy of chromosome 21. Mice engineered to model this aneuploidy exhibit Down syndrome-like memory deficits in spatial and contextual tasks. While abnormal neuronal function has been identified in these models, most studies have relied on in vitro measures. Here, using in vivo recording in the Dp(16)1Yey model, we find alterations in the organization of spiking of hippocampal CA1 pyramidal neurons, including deficits in the generation of complex spikes. These changes lead to poorer spatial coding during exploration and less coordinated activity during sharp-wave ripples, events involved in memory consolidation. Further, the density of CA1 inhibitory neurons expressing neuropeptide Y, a population key for the generation of pyramidal cell bursts, were significantly increased in Dp(16)1Yey mice. Our data refine the 'over-suppression' theory of Down syndrome pathophysiology and suggest specific neuronal subtypes involved in hippocampal dysfunction in these model mice.


Assuntos
Região CA1 Hipocampal/patologia , Região CA1 Hipocampal/fisiopatologia , Síndrome de Down/patologia , Síndrome de Down/fisiopatologia , Memória , Potenciais de Ação , Animais , Região CA1 Hipocampal/química , Modelos Animais de Doenças , Camundongos , Neuropeptídeo Y/análise
11.
J Surg Res ; 223: 230-236, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29433879

RESUMO

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) is among the most dismal of human malignancies. Neuropeptides have shown to be implicated in angiogenesis, tumor growth, and formation of distant metastases in various solid tumors. In the present study, we used a genetically engineered mouse model of pancreatic cancer to evaluate the impact of neuropeptide Y (NPY) and its receptors 1 (Y1) and 2 (Y2) in preneoplastic lesions and pancreatic cancer as a potential target with antiproliferative properties. In addition, human PDAC tissue was analyzed. MATERIALS AND METHODS: By interbreeding conditional LsL-Trp53R172H,LsL-KrasG12D and Pdx1-Cre strains, we obtained LsL-KrasG12D;LsL-Trp53R172H;Pdx1-Cre(KPC), LsL-KrasG12D;Pdx1-Cre(KP) and control mice (n = 8 each). Mice were then followed in a longitudinal study for 3 to 6 mo. Pancreata were analyzed in regard to pancreatic intraepithelial neoplasia (PanIN) lesions and invasive carcinoma. Corresponding sections were then assessed by immunohistochemistry and quantitative polymerase chain reaction for NPY, Y1 and Y2 expression in murine and human samples. RESULTS: NPY and Y1 expressions were detected in human and murine pancreatic samples, but expression levels were similar in neoplastic and non-neoplastic tissue. Y2 revealed a significant increase of expression in the transgenic mouse model in PanIN lesions and pancreatic cancer compared to control. This holds also true for human samples of pancreatic cancer. Immunohistochemistry of Y2 in murine and human samples of PanINs and pancreatic carcinoma revealed an increased expression in PanIN lesions and pancreatic cancer. CONCLUSIONS: Y2 is strongly overexpressed in pancreatic cancer and may modulate angiogenesis.


Assuntos
Carcinoma in Situ/química , Neuropeptídeo Y/análise , Neoplasias Pancreáticas/química , Receptores de Neuropeptídeo Y/análise , Animais , Humanos , Imuno-Histoquímica , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Pâncreas/química
12.
Sci Rep ; 7(1): 17984, 2017 12 21.
Artigo em Inglês | MEDLINE | ID: mdl-29269733

RESUMO

The ability to regulate food intake is critical to survival. The hypothalamus is central to this regulation, integrating peripheral signals of energy availability. Although our understanding of hunger in rodents is advanced, an equivalent understanding in birds is lacking. In particular, the relationship between peripheral energy indices and hypothalamic 'hunger' peptides, agouti-related protein (AgRP), pro-opiomelanocortin (POMC) and neuropeptide Y (NPY) is poorly understood. Here, we compare AgRP, POMC and NPY RNA levels in the hypothalamus of Red Junglefowl chicks raised under ad libitum, chronic restriction and intermittent feeding regimens. Hypothalamic gene expression differed between chronically and intermittently restricted birds, confirming that different restriction regimens elicit different patterns of hunger. By assessing the relationship between hypothalamic gene expression and carcass traits, we show for the first time in birds that AgRP and POMC are responsive to fat-related measures and therefore represent long-term energy status. Chronically restricted birds, having lower indices of fat, show elevated hunger according to AgRP and POMC. NPY was elevated in intermittently fasted birds during fasting, suggesting a role as a short-term index of hunger. The different physiological and neuroendocrine responses to quantitative versus temporal feed restriction provide novel insights into the divergent roles of avian hunger neuropeptides.


Assuntos
Galinhas/fisiologia , Fome/fisiologia , Proteína Relacionada com Agouti/análise , Proteína Relacionada com Agouti/fisiologia , Animais , Metabolismo Energético/fisiologia , Feminino , Privação de Alimentos/fisiologia , Hipotálamo/química , Hipotálamo/fisiologia , Masculino , Neuropeptídeo Y/análise , Neuropeptídeo Y/fisiologia , Pró-Opiomelanocortina/análise , Pró-Opiomelanocortina/fisiologia
13.
Explore (NY) ; 13(5): 306-312, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28915981

RESUMO

OBJECTIVES: To explore whether transcutaneous electrical acupoint stimulation (TEAS) can improve the outcomes of in vitro fertilization (IVF). DESIGN: A prospective, randomized, and controlled study. SETTING: IVF center in a university hospital. PARTICIPANTS: Four hundred and eighty-one infertile patients with bilateral tubal blockage who were referred for IVF. Patients were randomized into four groups. INTERVENTION: TEAS was administered for 30min, respectively, at 24h before TVOR and two hours before ET. The acupoints included SP10 (Xuehai, bilateral), SP8 (Diji, bilateral), LR3 (Taichong, bilateral), ST36 (Zusanli, bilateral), EX-CA1 (Zigong, bilateral), RN4 (Guanyuan), PC6 (Neiguan, bilateral), and RN12 (Zhongwan). Based on different frequencies of TEAS, patients were grouped into a TEAS-2Hz group, a TEAS-100Hz group and a TEAS-2/100Hz group. Patients in the control group only received routine IVF treatment and no TEAS was applied on them. PRIMARY AND SECONDARY OUTCOME MEASURES: The number of mature oocytes, normally fertilized oocytes and good-quality embryos were used to evaluate oocyte developmental competence of the patients. Data of clinical pregnancy rate (CPR), implantation rate (IR), and live birth rate (LBR) were also obtained. The levels of neuropeptide Y (NPY), transforming growth factor alpha and granulocyte colony-stimulating factor in the follicular fluids were measured with enzyme-linked immunosorbent assay (ELISA). RESULTS: No significant differences were found between the control, TEAS-2Hz, TEAS-100Hz and TEAS-2/100Hz groups on the numbers of metaphase II oocytes, normally fertilized zygotes, early cleavage embryos or good quality embryos (P > .05). However, the CPR, IR and LBR of the TEAS-2/100Hz group were significantly higher than those of the other groups, respectively (P < .05). The NPY levels in the follicular fluids of TEAS-2/100Hz group were significantly higher than those of the other groups (P < .05). CONCLUSION: TEAS using a frequency of 2/100Hz could help to improve the IVF outcomes partly by increasing NPY levels in the follicular fluids.


Assuntos
Eletroacupuntura/métodos , Fertilização In Vitro , Infertilidade Feminina/terapia , Estimulação Elétrica Nervosa Transcutânea , Adulto , China , Feminino , Líquido Folicular/química , Fator Estimulador de Colônias de Granulócitos/análise , Humanos , Neuropeptídeo Y/análise , Oócitos/fisiologia , Gravidez , Taxa de Gravidez , Estudos Prospectivos , Fator de Crescimento Transformador alfa/análise , Resultado do Tratamento
14.
High Alt Med Biol ; 18(3): 234-241, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28486037

RESUMO

Liu, Xiang-Wen, Jie Yin, Qi-Sheng Ma, Chu-Chu Qi, Ji-Ying Mu, Lang Zhang, Li-Ping Gao, and Yu-Hong Jing. Role of arcuate nucleus in the regulation of feeding behavior in the process of altitude acclimatization in rats. High Alt Med Biol. 18:234-241, 2017.-Highly efficient energy utilization and metabolic homeostasis maintenance rely on neuromodulation. Altitude exposure is known to stimulate neuroendocrine systems to respond to acute hypoxia and adaptive acclimatization. However, limited data on how the adaptive regulation of the arcuate nucleus performs in the process of altitude acclimatization are available. In the present study, male Sprague Dawley rats were transported to Huashixia, Qinghai (with an altitude of 4400 m) from Xian (with an altitude of 300 m) by air; rats were consistently raised in Xian as control. Food uptake and body weight were measured consecutively after being subjected to high-altitude condition. Contents of plasma leptin and ghrelin were analyzed by the Enzyme Linked Immunosorbent Assay (ELISA) Kits. Brain coronal sections were obtained, and neuropeptide Y (NPY), proopiomelanocotin (POMC), and c-fos immunoreactivity in arcuate nucleus were observed. Arcuate nucleus was isolated from the hypothalamus, and the mRNA of NPY and POMC were measured by quantitative real-time polymerase chain reaction. Our results showed both food consumption and body weight decreased in the high plateau compared with rats raised in the low-altitude condition. Plasma leptin increased at the early stage, and ghrelin decreased at a later stage after reaching the high plateau. The peak of c-fos immunoreactivity in the arcuate nucleus was at day 3 after reaching the high plateau. The expression level of NPY increased, and POMC decreased in the arcuate nucleus at day 7 after reaching the high plateau compared with the plain control group. These results indicate that the arcuate nucleus of hypothalamus performs an important function in regulating feeding behavior during altitude acclimatization. Our study suggested that altitude acclimation is regulated by the hypothalamus that received leptin and ghrelin signals to response by its microcircuit, including NPY- and POMC-neurons in the arcuate nucleus.


Assuntos
Aclimatação/fisiologia , Doença da Altitude/fisiopatologia , Altitude , Núcleo Arqueado do Hipotálamo/fisiopatologia , Comportamento Alimentar/fisiologia , Animais , Peso Corporal , China , Genes fos/fisiologia , Grelina/sangue , Hipotálamo/fisiopatologia , Leptina/sangue , Masculino , Neuropeptídeo Y/análise , Pró-Opiomelanocortina/análise , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley
15.
J Endocrinol ; 234(1): 41-56, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28455431

RESUMO

Early life diet influences metabolic programming, increasing the risk for long-lasting metabolic ill health. Neonatally overfed rats have an early increase in leptin that is maintained long term and is associated with a corresponding elevation in body weight. However, the immediate and long-term effects of neonatal overfeeding on hypothalamic anorexigenic pro-opiomelanocortin (POMC) and orexigenic agouti-related peptide (AgRP)/neuropeptide Y (NPY) circuitry, and if these are directly mediated by leptin, have not yet been examined. Here, we examined the effects of neonatal overfeeding on leptin-mediated development of hypothalamic POMC and AgRP/NPY neurons and whether these effects can be normalised by neonatal leptin antagonism in male Wistar rats. Neonatal overfeeding led to an acute (neonatal) resistance of hypothalamic neurons to exogenous leptin, but this leptin resistance was resolved by adulthood. While there were no effects of neonatal overfeeding on POMC immunoreactivity in neonates or adults, the neonatal overfeeding-induced early increase in arcuate nucleus (ARC) AgRP/NPY fibres was reversed by adulthood so that neonatally overfed adults had reduced NPY immunoreactivity in the ARC compared with controls, with no further differences in AgRP immunoreactivity. Short-term neonatal leptin antagonism did not reverse the excess body weight or hyperleptinaemia in the neonatally overfed, suggesting factors other than leptin may also contribute to the phenotype. Our findings show that changes in the availability of leptin during early life period influence the development of hypothalamic connectivity short term, but this is partly resolved by adulthood indicating an adaptation to the metabolic mal-programming effects of neonatal overfeeding.


Assuntos
Animais Recém-Nascidos/fisiologia , Dieta , Hipotálamo/fisiologia , Leptina/fisiologia , Hipernutrição , Proteína Relacionada com Agouti/análise , Proteína Relacionada com Agouti/fisiologia , Animais , Núcleo Arqueado do Hipotálamo/química , Resistência a Medicamentos , Feminino , Hipotálamo/química , Leptina/antagonistas & inibidores , Leptina/farmacologia , Tamanho da Ninhada de Vivíparos , Masculino , Neurônios/fisiologia , Neuropeptídeo Y/análise , Neuropeptídeo Y/fisiologia , Pró-Opiomelanocortina/análise , Pró-Opiomelanocortina/fisiologia , Ratos , Ratos Wistar
16.
J Comp Neurol ; 525(10): 2394-2410, 2017 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-28324630

RESUMO

The vagina is innervated by a complex arrangement of sensory, sympathetic, and parasympathetic nerve fibers that contain classical transmitters plus an array of neuropeptides and enzymes known to regulate diverse processes including blood flow and nociception. The neurochemical characteristics and distributions of peptide-containing nerves in the mouse vagina are unknown. This study used multiple labeling immunohistochemistry, confocal maging and analysis to investigate the presence and colocalization of the peptides vasoactive intestinal polypeptide (VIP), calcitonin-gene related peptide (CGRP), substance P (SP), neuropeptide tyrosine (NPY), and the nitric oxide synthesizing enzyme neuronal nitric oxide synthase (nNOS) in nerve fibers of the murine vaginal wall. We compared cervical and vulvar areas of the vagina in young nullipara and older multipara C57Bl/6 mice, and identified differences including that small ganglia were restricted to cervical segments, epithelial fibers were mainly present in vulvar segments and most nerve fibers were found in the lamina propria of the cervical region of the vagina, where a higher number of fibers containing immunoreactivity for VIP, CGRP, SP, or nNOS were found. Two populations of VIP-containing fibers were identified: fibers containing CGRP and fibers containing VIP but not CGRP. Differences between young and older mice were present in multiple layers of the vaginal wall, with older mice showing overall loss of innervation of epithelium of the proximal vagina and reduced proportions of VIP, CGRP, and SP containing nerve fibers in the distal epithelium. The distal vagina also showed increased vascularization and perivascular fibers containing NPY. Immunolabeling of ganglia associated with the vagina indicated the likely origin of some peptidergic fibers. Our results reveal regional differences and age- or parity-related changes in innervation of the mouse vagina, effecting the distribution of neuropeptides with diverse roles in function of the female genital tract.


Assuntos
Peptídeo Relacionado com Gene de Calcitonina/análise , Fibras Nervosas/química , Neuropeptídeo Y/análise , Substância P/análise , Vagina/química , Peptídeo Intestinal Vasoativo/análise , Animais , Peptídeo Relacionado com Gene de Calcitonina/metabolismo , Feminino , Camundongos , Camundongos Endogâmicos C57BL , Fibras Nervosas/metabolismo , Neuropeptídeo Y/metabolismo , Óxido Nítrico Sintase Tipo I/análise , Óxido Nítrico Sintase Tipo I/metabolismo , Substância P/metabolismo , Vagina/citologia , Vagina/metabolismo , Peptídeo Intestinal Vasoativo/metabolismo
17.
Wound Repair Regen ; 25(1): 75-85, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27997734

RESUMO

Blood supply is the most important factor determining the survival of a skin flap. Botulinum toxin-A (Botox-A) is used as pharmacologic agent not only for aesthetic purposes, but also for its vasomotor actions. This study was conducted to establish whether local application of Botox-A increased survival of random pattern skin flaps in rats by changing the expression of neurotransmitters. Forty adult Sprague-Dawley rats with a caudally-based random pattern skin flap were divided into two groups: Botox-A group and saline group. Surviving flap area and cutaneous blood flow in the flap were evaluated on postoperative days 3 and 7. After injection of Botox-A, changes in vessels were analyzed using immunohistochemical staining. Levels of norepinephrine, neuropeptide-Y, nitric oxide, and endothelial nitric oxide synthase were analyzed quantitatively by high performance liquid chromatography, Western blot, and colorimetric assay. The survived area in the Botox-A group was significantly higher than that in the control group on postoperative days 3 and 7. Blood flow in the Botox-A group was significantly high in the proximal and middle areas immediately after the operation. The number of CD31-positive vessels in the Botox-A group was significant greater than that in the control group. Norepinephrine level in the Botox-A group decreased significantly immediately after flap elevation and at postoperative day 3. There were no significant differences in neuropeptide-Y level between the two groups. Nitric oxide level did not change significantly in either group despite the increase in endothelial nitric oxide synthase immediately after flap elevation and at 3 days postoperatively. In conclusion, Botox-A increased vascular blood flow and viable flap area in rats by reducing norepinephrine level. In contrast, neuropeptide-Y, another vasoconstrictor, was not affected by Botox-A. Nitric oxide, a vasodilator, was also not affected by Botox-A, despite the significant increase in endothelial nitric oxide synthase expression in the flaps.


Assuntos
Toxinas Botulínicas Tipo A/administração & dosagem , Fármacos Cardiovasculares/administração & dosagem , Neurotransmissores/biossíntese , Pele/efeitos dos fármacos , Retalhos Cirúrgicos/irrigação sanguínea , Cicatrização/efeitos dos fármacos , Animais , Modelos Animais de Doenças , Imuno-Histoquímica , Injeções , Neuropeptídeo Y/análise , Neuropeptídeo Y/metabolismo , Óxido Nítrico/análise , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo III/análise , Óxido Nítrico Sintase Tipo III/metabolismo , Norepinefrina/análise , Norepinefrina/metabolismo , Ratos , Ratos Sprague-Dawley , Pele/irrigação sanguínea , Pele/fisiopatologia , Simpatomiméticos/metabolismo , Vasoconstrição/efeitos dos fármacos , Vasoconstrição/fisiologia , Cicatrização/fisiologia
18.
J Eur Acad Dermatol Venereol ; 31(3): 443-449, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27427400

RESUMO

BACKGROUND: Neuropeptide Y (NPY) is involved in the carcinogenesis of different tumours, especially neural crest-derived tumours. OBJECTIVE: The aim of our study is to investigate the expression of NPY on melanoma and its relation with prognostic histological parameters and survival. METHODS: This is a retrospective observational study of two independent series, with a total of 79 primary melanomas, diagnosed in two independent University Hospitals in Spain, from January 2000 to December 2004. RESULTS: We found a significant higher expression of NPY on superficial spreading melanoma and lentigo maligna (40%) (P = 0.030). Thinner tumours were associated with higher NPY expression (Clark level, P = 0.003; Breslow level, P = 0.012). Melanomas with low NPY expression were associated with intense cell proliferation (Ki-67, P = 0.034), high density of peritumoral mast cell infiltrates (P = 0.033) and low E-cadherin expression (P = 0.031). Melanomas with high NPY expression exhibited significant differences in terms of relapse time (median: 114 vs. 68 months, P = 0.008) and overall survival (114 vs. 74 months, P = 0.004). CONCLUSION: High expression of NPY was associated with better prognostic histological parameters, low peritumoral mast cells density, presence of adhesion proteins and better outcome.


Assuntos
Melanoma/química , Neuropeptídeo Y/análise , Neoplasias Cutâneas/química , Adulto , Idoso , Idoso de 80 Anos ou mais , Caderinas/análise , Proliferação de Células , Intervalo Livre de Doença , Feminino , Humanos , Sarda Melanótica de Hutchinson/química , Sarda Melanótica de Hutchinson/patologia , Antígeno Ki-67/análise , Masculino , Mastócitos , Melanoma/patologia , Pessoa de Meia-Idade , Estudos Retrospectivos , Neoplasias Cutâneas/patologia , Taxa de Sobrevida , Carga Tumoral
19.
PLoS One ; 11(11): e0165465, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27802308

RESUMO

Bone repair is a specialized type of wound repair controlled by complex multi-factorial events. The nervous system is recognized as one of the key regulators of bone mass, thereby suggesting a role for neuronal pathways in bone homeostasis. However, in the context of bone injury and repair, little is known on the interplay between the nervous system and bone. Here, we addressed the neuropeptide Y (NPY) neuronal arm during the initial stages of bone repair encompassing the inflammatory response and ossification phases in femoral-defect mouse model. Spatial and temporal analysis of transcriptional and protein levels of NPY and its receptors, Y1R and Y2R, reported to be involved in bone homeostasis, was performed in bone, dorsal root ganglia (DRG) and hypothalamus after femoral injury. The results showed that NPY system activity is increased in a time- and space-dependent manner during bone repair. Y1R expression was trigged in both bone and DRG throughout the inflammatory phase, while a Y2R response was restricted to the hypothalamus and at a later stage, during the ossification step. Our results provide new insights into the involvement of NPY neuronal pathways in bone repair.


Assuntos
Fêmur/lesões , Hipotálamo/fisiologia , Vias Neurais/fisiologia , Neuropeptídeo Y/metabolismo , Cicatrização , Animais , Fêmur/inervação , Fêmur/patologia , Fêmur/fisiologia , Gânglios Espinais/metabolismo , Gânglios Espinais/patologia , Regulação da Expressão Gênica , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neuropeptídeo Y/análise , Neuropeptídeo Y/genética , RNA Mensageiro/genética , Receptores de Neuropeptídeo Y/análise , Receptores de Neuropeptídeo Y/metabolismo , Transdução de Sinais
20.
Small ; 12(40): 5524-5529, 2016 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-27551968

RESUMO

Silicon nanowire field-effect transistors modified with specific aptamers can directly detect the minute dopamine and neuropeptide Y released from cells. The binding of these molecules to the aptamers results in a conductance change of the transistor biosensor and illustrates the differential releasing mechanisms of these molecules stored in various vesicle pools.


Assuntos
Aptâmeros de Peptídeos/química , Dopamina/análise , Histamina/farmacologia , Nanofios/química , Neuropeptídeo Y/análise , Transistores Eletrônicos , Animais , Células PC12 , Ratos , Silício/química
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