RESUMO
The mammalian oocyte is surrounded by a matrix called the zona pellucida (ZP). This envelope participates in processes such as acrosome reaction induction, sperm binding and may be involved in speciation. In cat (Felis catus), this matrix is composed of at least three glycoproteins called ZP2, ZP3, and ZP4. However, recent studies have pointed to the presence of a fourth protein in several mammals (rat, human, hamster or rabbit), meaning that a reevaluation of cat ZP is needed. For this reason, the objective of this research was to analyze the protein composition of cat ZP by means of proteomic analysis. Using ZP from ovaries and oocytes, several peptides corresponding to four proteins were detected, yielding a coverage of 33.17%, 71.50%, 50.23%, and 49.64% for ZP1, ZP2, ZP3, and ZP4, respectively. Moreover, the expression of four genes was confirmed by molecular analysis. Using total RNA isolated from cat ovaries, the complementary deoxyribonucleic acids encoding cat ZP were partially amplified by reverse-transcribed polymerase chain reaction. Furthermore, ZP1 was totally amplified for the first time in this species. As far as we are aware, this is the first study that confirms the presence of four proteins in cat ZP.
Assuntos
Gatos/genética , Proteínas do Ovo/análise , Proteínas do Ovo/genética , Expressão Gênica , Glicoproteínas de Membrana/análise , Glicoproteínas de Membrana/genética , Receptores de Superfície Celular/análise , Receptores de Superfície Celular/genética , Zona Pelúcida/metabolismo , Sequência de Aminoácidos , Animais , Proteínas do Ovo/química , Feminino , Glicoproteínas de Membrana/química , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Proteômica , RNA Mensageiro/análise , Receptores de Superfície Celular/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Zona Pelúcida/química , Glicoproteínas da Zona PelúcidaRESUMO
The pharmacokinetics of dexketoprofen trometamol were evaluated in two studies using healthy volunteers. In the first study, the relative bioavailability of a single oral capsule of dexketoprofen free acid 25 mg or dexketoprofen trometamol 25 mg (given as 37 mg of the trometamol salt) was compared to ketoprofen 50 mg in 18 healthy volunteers. In the second study, the pharmacokinetics and tolerability of oral dexketoprofen trometamol in tablet form were evaluated after either a single 25 mg dose (24 volunteers) or a repeated dose of 25 mg twice daily for 7 days (12 volunteers). The absorption of dexketoprofen from dexketoprofen trometamol capsules was bioequivalent to that of ketoprofen. On the other hand, the extent of absorption of dexketoprofen free acid was significantly lower than that for ketoprofen. Dexketoprofen trometamol showed the most rapid absorption rate, with highest Cmax and shortest t(max) values, whereas dexketoprofen free acid had the slowest absorption rate, and ketoprofen had an intermediate absorption rate. After repeated-dose administration of dexketoprofen trometamol, the pharmacokinetic parameters were similar to those obtained after single doses, indicating that no drug accumulation occurred. Dexketoprofen trometamol was well tolerated, with no clinically relevant adverse events reported.
Assuntos
Anti-Inflamatórios não Esteroides/farmacocinética , Cetoprofeno/análogos & derivados , Trometamina/análogos & derivados , Absorção , Administração Oral , Adulto , Anti-Inflamatórios não Esteroides/administração & dosagem , Anti-Inflamatórios não Esteroides/efeitos adversos , Disponibilidade Biológica , Estudos Cross-Over , Método Duplo-Cego , Feminino , Humanos , Cetoprofeno/farmacologia , Cetoprofeno/toxicidade , Masculino , Fatores Sexuais , Trometamina/farmacologia , Trometamina/toxicidadeRESUMO
The zona pellucida (ZP) participates in sperm-egg interactions during the first steps of fertilization. Recent studies have shown that the ZP matrix of oocytes in several species is composed of four glycoproteins, designated as ZP1, ZP2, ZP3 and ZP4, rather than the three described in mouse, pig and cow. In this study, investigations were carried out to unveil a fourth glycoprotein in the rabbit (Oryctolagus cuniculus) ZP. Using total RNA isolated from rabbit ovaries, the complementary deoxyribonucleic acid (cDNA) encoding rabbit ZP1 was amplified by reverse transcribed polymerase chain reaction (RT-PCR). The ZP1 cDNA contains an open reading frame of 1825 nucleotides encoding a polypeptide of 608 amino acid residues. The deduced amino acid sequence of rabbit ZP1 showed high identity with other species: 70% identity with human and horse ZP1, and 67% identity with mouse and rat ZP1. At the proteomic level, peptides corresponding to the four proteins were detected by mass spectrometry. In addition, a molecular phylogenetic analysis of ZP1 showed that pseudogenization of this gene has occurred at least four times during the evolution of mammals. The data presented in this manuscript provide evidence, for the first time, that the rabbit ZP is composed of four glycoproteins.
Assuntos
Proteínas do Ovo/análise , Glicoproteínas de Membrana/análise , Receptores de Superfície Celular/análise , Zona Pelúcida/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Cromatografia Líquida de Alta Pressão , Proteínas do Ovo/genética , Proteínas do Ovo/isolamento & purificação , Feminino , Glicoproteínas/análise , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/isolamento & purificação , Dados de Sequência Molecular , Filogenia , Proteômica , Pseudogenes/genética , Coelhos , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/isolamento & purificação , Alinhamento de Sequência , Espectrometria de Massas em Tandem , Glicoproteínas da Zona PelúcidaRESUMO
Captopril is an angiotensin-converting enzyme inhibitor (ACEI) used in the treatment of hypertension and congestive heart failure and has demonstrated its cardiovascular effects in experimental animal models, healthy volunteers and patients. The aim of this study was to find out whether or not differences in the pharmacokinetic profile and the haemodynamic response of a 100-mg single oral dose of captopril appeared between subjects of both sexes. Twenty-four young healthy volunteers (12 males and 12 females) took part in the trial. Blood samples to assess captopril plasma concentrations, determined by reverse phase high performance liquid chromatography (HPLC), as well as haemodynamic variables, were obtained before and at different times following drug intake. Pharmacokinetic parameters did not show significant sex differences. Systolic and diastolic blood pressure exhibited a statistically significant decrease between 0.5 and 8 h in both sexes. No significant sex-related differences were found. The drug exhibited a good tolerability.