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1.
Front Mol Neurosci ; 16: 1171855, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37251645

RESUMO

N-(3-fluoro-1-phenethylpiperidine-4-yl)-N-phenyl propionamide is a newly-designed pain killer selectively activating G-protein-coupled mu-opioid receptors (MOR) in acidic injured tissues, and therefore devoid of central side effects which are typically elicited at normal pH values in healthy tissues. However, the neuronal mechanisms underlying NFEPP's antinociceptive effects were not examined in detail so far. Voltage-dependent Ca2+ channels (VDCCs) in nociceptive neurons play a major role in the generation and inhibition of pain. In this study, we focused on the effects of NFEPP on calcium currents in rat dorsal root ganglion (DRG) neurons. The inhibitory role of the G-protein subunits Gi/o and Gßγ on VDCCs was investigated using the blockers pertussis toxin and gallein, respectively. GTPγS binding, calcium signals and MOR phosphorylation were also investigated. All experiments were performed at acidic and normal pH values using NFEPP in comparison to the conventional opioid agonist fentanyl. At low pH, NFEPP produced more efficient G-protein activation in transfected HEK293 cells and significantly reduced VDCCs in depolarized DRG neurons. The latter effect was mediated by Gßγ subunits, and NFEPP-mediated MOR phosphorylation was pH-dependent. Fentanyl's responses were not affected by pH changes. Our data indicate that NFEPP-induced MOR signaling is more effective at low pH and that the inhibition of calcium channels in DRG neurons underlies NFEPP's antinociceptive actions.

2.
Methods Mol Biol ; 2201: 83-95, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-32975791

RESUMO

Real-time quantitative reverse transcription-PCR (qRT-PCR ) is a highly sensitive molecular biology method based on the amplification of the cDNA of mRNA to detect and quantify the levels of mRNA of interest. In this chapter, we describe real-time qRT-PCR to detect and quantify mRNA of opioid receptors in immune cells. Specifically, we analyze mouse immune cells isolated from the blood and sciatic nerves exposed to a chronic constriction injury, which represents a model of neuropathic pain. We describe in detail the requirements and techniques to induce the chronic constriction injury, to isolate immune cells from the blood and injured nerves, to isolate the total RNA from immune cells, to perform a cDNA reverse transcription from the total RNA, and to perform real-time qRT-PCR for µ-, δ-, and κ-opioid receptor mRNAs.


Assuntos
Reação em Cadeia da Polimerase em Tempo Real/métodos , Receptores Opioides mu/genética , Animais , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Neuralgia , RNA Mensageiro/genética , Receptores Acoplados a Proteínas G/genética , Receptores Acoplados a Proteínas G/metabolismo , Receptores Opioides/genética , Receptores Opioides/imunologia , Receptores Opioides/metabolismo , Receptores Opioides delta/genética , Receptores Opioides kappa/genética , Receptores Opioides mu/imunologia , Receptores Opioides mu/metabolismo , Transcrição Reversa , Nervo Isquiático/lesões
3.
Ear Nose Throat J ; 95(3): E8-E11, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26991232

RESUMO

The aim of this study was to investigate the presence of Helicobacter pylori in human lacrimal and nasal secretions. Eighty patients with complaints of dyspepsia who had undergone endoscopies and gastric antrum biopsies were included in the study. A total of five specimens, including 2 lacrimal secretion samples, 2 nasal mucosal swab samples, and 1 gastric antrum biopsy, were collected from each patient and investigated with polymerase chain reaction (PCR) methods consisting of the urease enzyme coding gene GlmM (UreC) and the H pylori-specific 16S rRNA coding gene. The Reflux Symptom Index and ophthalmologic complaints of the patients were recorded. The detected positivity rates of the H pylori 16S rRNA coding gene in gastric biopsies and nasal mucous and lacrimal secretions were 55, 11.2, and 20%, respectively. The patients were grouped as gastric-antrum-biopsy-negative (Group I [n = 36]) and -positive (Group II [n = 44). In Group II, H pylori positivity in the lacrimal and nasal mucous secretions was 36.3 and 18%, respectively. A comparison between the groups in terms of H pylori presence in nasal mucous and lacrimal secretions yielded statistically significant differences (p = 0.0001, p = 0.003). The simultaneous presence of H pylori in nasal mucous and lacrimal secretions was 13.6% in Group II. H pylori positivity in nasal mucous and lacrimal secretions had a positive moderate correlation (r = 0.40; p = 0.0003). The present study is the first report on the presence of H pylori in lacrimal secretions through nested PCR, which suggested the presence of a number of mechanisms for H pylori transmission to lacrimal secretions.


Assuntos
Dispepsia/microbiologia , Infecções Oculares Bacterianas/microbiologia , Infecções por Helicobacter/microbiologia , Helicobacter pylori/isolamento & purificação , Lágrimas/microbiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , DNA Bacteriano/isolamento & purificação , Feminino , Mucosa Gástrica/microbiologia , Helicobacter pylori/genética , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa Nasal/microbiologia , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 16S/isolamento & purificação , Índice de Gravidade de Doença , Lágrimas/metabolismo , Adulto Jovem
4.
Arch Gynecol Obstet ; 294(3): 511-7, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-26781261

RESUMO

PURPOSE: To evaluate various aspects of two popular uterine sparing techniques, the B-Lynch uterine compression suture and Bakri balloon tamponade, in severe postpartum hemorrhage (PPH). METHODS: 21 women who underwent the Bakri balloon procedure and 24 women who underwent the B-Lynch suture as primary uterus-sparing methods, due to PPH not responding to medical treatment, were retrospectively evaluated. RESULTS: The success rates of the B-Lynch procedure and the Bakri balloon were 79.1 and 80 %, respectively. The success rates of the B-Lynch + IIAL and the Bakri balloon + IIAL were 91.6 and 95 %, respectively. There was no significant difference in success rates, mean duration of time to stop bleeding, estimated blood loss, transfused packed red blood cells or mean duration of hospital stay between the B-Lynch and the Bakri balloon groups. The duration of operation was significantly longer in the Bakri balloon compared to the B-Lynch group (p = 0.01). CONCLUSION: In our study, the Bakri balloon and the B-Lynch suture had similar success rates in uterine atony during CS. The advantages of the B-Lynch suture include rapid application with no need for lithotomy position or extra material; whereas the Bakri balloon is less invasive and easier to learn, but more time consuming and expensive compared to the B-Lynch suture. We suggest that the B-Lynch suture may be preferred in uterine atony during CS in low resource settings; however, the less invasive Bakri balloon should be the first line in full resource settings. Further studies are needed to evaluate the advantages and disadvantages of the two methods.


Assuntos
Cesárea , Técnicas de Sutura , Tamponamento com Balão Uterino , Inércia Uterina/cirurgia , Útero/cirurgia , Adulto , Feminino , Humanos , Gravidez , Estudos Retrospectivos
5.
Neuropharmacology ; 101: 330-40, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26453963

RESUMO

Targeting peripheral neuropathic pain at its origin may prevent the development of hypersensitivity. Recently we showed this can be mediated by opioid receptors at the injured nerve trunk. Here, we searched for the most relevant peripheral site to block transient receptor potential vanilloid 1 (TRPV1), and investigated analgesic interactions between TRPV1 and opioids in neuropathy. In a chronic constriction injury (CCI) of the sciatic nerve in mice, we assessed the effects of µ-, δ- and κ-opioid receptor agonists and TRPV1 antagonist (SB366791) injected at the CCI site or into the injured nerve-innervated paw on spontaneous paw lifting, heat and mechanical sensitivity. We also examined TRPV1 expression in total membrane and plasma membrane fractions from nerves and paws. We found that opioids and SB366791 co-injected in per se nonanalgesic doses at the CCI site or into the paw diminished heat and mechanical sensitivity. SB366791 alone dose-dependently alleviated heat and mechanical sensitivity. TRPV1 blockade in the paw was more effective than at the CCI site. None of the treatments diminished spontaneous paw lifting. TRPV1 expression analysis suggests that the levels of functional TRPV1 do not critically determine the TRPV1 antagonist-mediated analgesia. Together, the identification of the primary action site in damaged nerves is crucial for effective pain control. Contrary to opioids, the TRPV1 blockade in the injured nerve peripheral terminals, rather than at the nerve trunk, appears promising against heat pain. Opioid/TRPV1 antagonist combinations at both locations partially reduced neuropathy-triggered heat and mechanical pain.


Assuntos
Analgésicos Opioides/uso terapêutico , Neuralgia/tratamento farmacológico , Canais de Cátion TRPV/metabolismo , Análise de Variância , Anilidas/uso terapêutico , Animais , Proteínas de Arabidopsis , Cinamatos/uso terapêutico , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Combinação de Medicamentos , Ala(2)-MePhe(4)-Gly(5)-Encefalina/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Hiperalgesia/complicações , Hiperalgesia/tratamento farmacológico , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Nociceptividade/efeitos dos fármacos , Proteínas Nucleares , Medição da Dor/efeitos dos fármacos , Limiar da Dor/efeitos dos fármacos , Canais de Cátion TRPV/genética , Fatores de Tempo
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