RESUMO
Immune checkpoint therapy to reverse natural killer (NK) and T cell exhaustion has emerged as a promising treatment in various cancers. While anti-programmed cell death 1 (PD-1) pembrolizumab has recently gained Food and Drug Administration (FDA) approval for use in recurrent or metastatic cervical cancer, other checkpoint molecules, such as T cell immunoreceptor with immunoglobulin (Ig) and immunoreceptor tyrosine-based inhibition motif (ITIM) domains (TIGIT) and T cell immunoglobulin and mucin-domain containing-3 (Tim-3), have yet to be fully explored in this disease. We report expression of TIGIT, Tim-3 and PD-1 on subsets of peripheral blood NK (CD56dim/neg CD16bright/dim/neg and CD56bright CD16dim/neg ) and T cells. The percentages of these cells were increased in women with cervical cancer and pre-malignant lesions. PD-1+ NK and T cells were likely to co-express TIGIT and/or Tim-3. These cells, with an apparently 'exhausted' phenotype, were augmented in patients. A subset of cells were also natural killer group 2 member D (NKG2D)- and DNAX accessory molecule 1 (DNAM-1)-positive. PD-1int and PD-1high T cells were notably increased in cervical cancer. Soluble programmed cell death ligand 1 (PD-L1) was higher in cancer patient blood versus healthy donors and we observed a positive correlation between sPD-L1 and PD-1+ T cells in women with low-grade lesions. Within the cancer group, there were no significant correlations between sPD-L1 levels and cervical cancer stage. However, when comparing cancer versus healthy donors, we observed an inverse association between sPD-L1 and total T cells and a correlation between sPD-L1 and CD56dim NK cells. Our results may show an overview of the immune response towards pre-cancerous lesions and cervical cancer, perhaps giving an early clue as to whom to administer blocking therapies. The increase of multiple checkpoint markers may aid in identifying patients uniquely responsive to combined antibody therapies.
Assuntos
Antígeno B7-H1/metabolismo , Células Matadoras Naturais/metabolismo , Receptor de Morte Celular Programada 1/metabolismo , Linfócitos T/metabolismo , Neoplasias do Colo do Útero/metabolismo , Adulto , Antígenos de Diferenciação de Linfócitos T/metabolismo , Antígeno CD56/metabolismo , Feminino , Citometria de Fluxo/métodos , Receptor Celular 2 do Vírus da Hepatite A/metabolismo , Humanos , Células Matadoras Naturais/imunologia , Pessoa de Meia-Idade , Subfamília K de Receptores Semelhantes a Lectina de Células NK/metabolismo , Receptores Imunológicos/metabolismo , Linfócitos T/imunologia , Neoplasias do Colo do Útero/imunologia , Neoplasias do Colo do Útero/patologia , Adulto JovemRESUMO
INTRODUCTION: Childhood obesity is a public health problem characterized by early insulin resistance (IR), inflammation, and oxidative stress. The presence of an uninterrupted low-grade inflammatory state impairs metabolic and cardiovascular health. The population is particularly susceptible to develop metabolic disorders related to increased body fat. METHODS: Eighty-three adolescents were recruited and grouped according to HOMA-IR and BMI in either with or without IR and obese or normal-weight respectively. Anthropometric, biochemical, immunological and hormonal variables were determined. Transverse Analytical Study. RESULTS: Obesity, dyslipidemia, IL-6, and C-reactive protein were significantly higher in the IR group than in the non-IR group. Obese adolescents showed increased insulin levels, HOMA-IR, inflammatory markers, and triglycerides; while having lower HDL-C, and adiponectin when compared to normal-weight adolescents. As expected, obesity-related anthropometric markers positively correlated with IR and inflammatory markers while negatively correlated with adiponectin levels. CONCLUSIONS: Early IR, subclinical inflammation, dyslipidemia, and hypoadiponectinemia characterize obesity in adolescents. These factors may increase the risk of future coronary heart disease (CHD) and diabetes mellitus development (DM) in early adulthood.
RESUMO
An aqueous extract of Casimiroa edulis leaves was tested in adult male Wistar rats for anticonvulsant activity utilizing two models of experimental epilepsy: maximal electroshock seizure (MES) and subcutaneously injected metrazole (METsc). Single dose of 100 mg/kg C. edulis vacuum dried aqueous extracts (VDA) orally administered to experimental animals elicited 50% and 70% abolition of MES and METsc-induced seizures, respectively. Two firmly established antiepileptic drugs in human therapy, phenytoin (PHT) and phenobarbital (PB), abolished 90% of MES-induced seizures, whereas an 80% and 100% absence of clonic seizures was attained in METsc test, correspondingly. The seizure abolition observed in C. edulis VDA treated rats was comparable with the anticonvulsive pattern exhibited by PHT and PB. These results suggest that potencially antiepileptic compounds are present in C. edulis extracts that deserve the study of their identity and mechanism of action.
Assuntos
Anticonvulsivantes/uso terapêutico , Fenobarbital/uso terapêutico , Fenitoína/uso terapêutico , Extratos Vegetais/uso terapêutico , Convulsões/tratamento farmacológico , Administração Oral , Animais , Anticonvulsivantes/administração & dosagem , Modelos Animais de Doenças , Eletrochoque , Injeções Subcutâneas , Masculino , México , Pentilenotetrazol/administração & dosagem , Pentilenotetrazol/toxicidade , Fenobarbital/administração & dosagem , Fenitoína/administração & dosagem , Extratos Vegetais/administração & dosagem , Folhas de Planta , Ratos , Ratos Wistar , Convulsões/induzido quimicamenteRESUMO
The ethyl ether (EE) and hydroalcoholic extract (HE) of Magnolia grandiflora L. (Magnoliaceae) seeds, a popular plant utilized in the Mexican traditional medicine because of its antispasmodic as well as other reported pharmacological effects, were studied in adult male Wistar rats. EE and HE orally administered in a single dose of 250 mg/kg (calculated on lipidic base) and 200 mg/kg, exhibited abolition of the extensor reflex of maximal electric induced-seizure test in 50 and 40% of the experimental animals, respectively. They significantly prolonged the sleeping time induced by pentobarbital and only the ethanol extract induced hypothermia. No neurological deficit was exhibited by either extract according to the gait, stance and righting test. Although ulterior toxicological and pharmacological insight is necessary, these results suggest that the chemical constituents of this plant could have utility in the control of epileptic patients presenting convulsive seizures.
Assuntos
Anticonvulsivantes/farmacologia , Plantas Medicinais/química , Animais , Anticonvulsivantes/toxicidade , Temperatura Corporal/efeitos dos fármacos , Eletrochoque , Etanol , Éter , Hipnóticos e Sedativos/farmacologia , Masculino , Medicina Tradicional , México , Pentobarbital/farmacologia , Extratos Vegetais/farmacologia , Extratos Vegetais/toxicidade , Ratos , Ratos Wistar , Sementes/química , Convulsões/prevenção & controle , Sono/efeitos dos fármacos , SolventesRESUMO
The genotypes and subtypes of 15 Mexican hepatitis B virus strains were determined by sequencing and phylogenetic analysis of the small S-gene. The most predominant strains were found to be divergent genotype/subtype F/adw4 strains (66.6%), followed by A/adw2 (20.0%), D/ayw3 (6.7%), and G/adw2 (6.7%). The S-genes of the Mexican genotype F strains and two Nicaraguan strains described previously formed a subcluster with more than 4% divergence from the other strains within this genotype. The Mexican strains within genotypes A and D showed the highest homology with strains from Europe and the United States. Ten amino acid substitutions not described previously were found in the S-genes of strains from nine chronic carriers, whereas the S gene in strains from six acute hepatitis B patients were highly conserved as compared to their respective genotypes. One genotype F strain from an HBsAg positive chronic carrier had a T to A mutation at position 647, forming a translational stop at codon 216. Two genotype F strains from HBsAg negative chronic carriers had a Val180 instead of an Ala found in the other genotype F strains. This study shows that a divergent genotype F predominates in Mexican strains analyzed, which presented amino acid substitutions not reported previously outside the a determinant.
Assuntos
Variação Genética , Antígenos de Superfície da Hepatite B/genética , Hepatite B/virologia , Sequência de Aminoácidos , Substituição de Aminoácidos , Sequência de Bases , DNA Viral/análise , Feminino , Genótipo , Vírus da Hepatite B/classificação , Vírus da Hepatite B/genética , Humanos , Masculino , México , Dados de Sequência Molecular , Reação em Cadeia da Polimerase/métodos , Homologia de Sequência de AminoácidosRESUMO
A study with valproic acid (VPA) was performed in Sprague-Dawley rats with the aim of relating drug serum levels to variations of leukocytes, platelets, coagulation time and other haematological parameters. VPA, 20 and 240 mg/kg, were administered orally, three times a day in groups of rats of both sexes throughout 7, 14, 30, 60 and 90 day periods. Total blood leukocytes averaged 7296 +/- 443 in control as well as in treated groups. Lymphocytes and monocytes decreased in all five groups when receiving the TD50 of 240 mg/kg (P less than 0.05). The VPA fasting serum levels were detectable if administered at 8 hr intervals. Nevertheless, VPA caused modification of the chosen parameters. VPA t1/2 averaged 3 hr in rats. Thus, even if the VPA effects were observed in our experiments, we suggest dividing the whole VPA dose into eight portions to be administered at 3 hr intervals in rats to attain constant serum VPA levels as well as constant drug effects.
Assuntos
Plaquetas/efeitos dos fármacos , Leucócitos/efeitos dos fármacos , Ácido Valproico/farmacologia , Animais , Coagulação Sanguínea/efeitos dos fármacos , Feminino , Hemoglobinas/metabolismo , Contagem de Leucócitos/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Masculino , Monócitos/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Contagem de Plaquetas/efeitos dos fármacos , Ratos , Ratos Endogâmicos , Ácido Valproico/sangueRESUMO
Excised cornea from adult New Zealand rabbits were incubated with progesterone-4-14C in Eagle's media for 96 hr. Samples were inactivated at intervals of 24 hr incubation periods. The following metabolites of progesterone were isolated: 20 alpha-Hydroxy-4-pregnen-3-one, 20-hydroxy-4-pregnen-3-one, 5 alpha-pregnane-3,20-dione; 5 beta-pregnane-3,20-dione and 6 beta-hydroxy-4-pregnen-3,20-dione. 20 alpha-Hydroxy-pregnen-3-one was the predominant metabolite of progesterone-4-14C. A linear increase was observed throughout 96 hr. The opposite was found for 5 alpha and 5 beta pregnane-3,20-dione. Compounds remaining at the origin of the paper chromatograms contained 6 beta-hydroxy-4-pregnen-3,20-dione and other still unidentified metabolites of progesterone-4-14C. Presence of 20 alpha and 20 beta-reductase; 5 alpha and 5 beta-reductase and 6 beta-hydroxylase enzyme systems are involved in corneal progesterone metabolism. No fungal neither bacterial enzymatic biotransformation occurred in the culture media.
Assuntos
Córnea/metabolismo , Progesterona/metabolismo , Animais , Radioisótopos de Carbono , Cromatografia em Camada Fina , Técnicas In Vitro , Cinética , Progesterona/análogos & derivados , CoelhosRESUMO
1. Phenytoin was intraperitoneally injected in six different experimental groups of adult Mongolian gerbils of both sexes at doses of 5, 100 and 150 mg/kg of body wt during 7 and 14 days. 2. Their brain, cerebellum, liver and kidney DNA, RNA and protein content were measured by conventional procedures. 3. The reference values of these rodents' cellular DNA were calculated through hepatocytes isolated following an intracardiac perfusion with collagenase in Hanks solution. 4. A concentration of 7.2 micrograms of DNA per 10(6) cells was found. 5. Phenytoin did not affect tissues of groups of animals receiving daily doses of 5 mg/kg of weight during 7 and 14 days of treatment; although a slight increase of protein concentration was observed in liver (P less than 0.01) and kidney (P less than 0.05). 6. RNA values were found to be significantly decreased in cerebellum (P less than 0.001), brain (P less than 0.05) and kidney (P less than 0.05) of those animals receiving the drug throughout 14 days. 7. DNA concentration was found to be decreased in all four tissues obtained from animals treated with daily doses of phenytoin of 100 (P less than 0.001) and 150 (P less than 0.001) mg/kg of body wt throughout 14 days. 8. These results indicate that phenytoin affected tissue cellularity only when administered at high doses and 14 days of interval of drug administration (P less than 0.001).
Assuntos
Substâncias Macromoleculares , Fenitoína/farmacologia , Animais , Peso Corporal/efeitos dos fármacos , Química Encefálica/efeitos dos fármacos , DNA/metabolismo , Feminino , Gerbillinae , Fígado/metabolismo , Masculino , Proteínas/metabolismo , RNA/metabolismoRESUMO
Phenytoin and its vehicle were orally administered to adult Sprague-Dawley rats during 7, 14 and 30 days at doses of 300 and 450 mg/kg/24 hr., respectively. We found: 1) Increased liver DNA concentration in subgroups of animals treated with 450 mg at 7 (P < 0.02) and 15 days (P < 0.001) Phenytoin serum levels were 19 ug/ml. 2) Increased protein concentration with 300 mg at 7 (P < 0.01) and 15 days (P < 0.001), respectively. 3) Cloudy swelling, vacuolar degeneration, liver sinusoids disappearance and lymphocytic cells infiltrate in subgroups of rats receiving vehicle throughout 6, 14 and 15 days correspondingly. The former lesion was found in all subgroups, except that 450 mg treated animals liver more severely affected. 4) Increased DNA concentration in kidney of subgroups receiving 450 mg/kg throughout 7 (P < 0.05), 15 (P < 0.001) and 30 days (P < 0.001), correspondingly. 5) Increased protein concentration in rats receiving 450 mg during 15 days (P < 0.001) and severely decreased at 30 days period. 6) Cloudy swelling was found in all treated animals subgroups. Seven cellular and tissue lesions were caused by vehicle at 15 and 30 days periods. 450 mg of phenytoin predominantly caused tissue condensation and vacuolar degeneration in kidney cortex. 7) propylene glycol do affect liver and kidney at doses below TD-50. Phenytoin stimulate kidney and liver cell proliferation. Caution should be observed when using parenteral phenytoin.
Assuntos
Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fenitoína/toxicidade , Administração Oral , Animais , Divisão Celular/efeitos dos fármacos , Doença Hepática Induzida por Substâncias e Drogas/etiologia , Doença Hepática Induzida por Substâncias e Drogas/patologia , DNA/biossíntese , Relação Dose-Resposta a Droga , Fígado Gorduroso/induzido quimicamente , Fígado Gorduroso/patologia , Feminino , Rim/metabolismo , Rim/patologia , Nefropatias/induzido quimicamente , Nefropatias/patologia , Fígado/metabolismo , Fígado/patologia , Masculino , Fenitoína/administração & dosagem , Fenitoína/farmacocinética , Propilenoglicol , Propilenoglicóis/toxicidade , Biossíntese de Proteínas , Ratos , Ratos Sprague-DawleyRESUMO
We investigated the effect of beta 3-adrenergic receptor (beta(3)AR) polymorphism on lipid profiles in patients with rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) treated with chloroquine. One hundred sixty-eight subjects were classified into three groups: 61 RA patients, 57 SLE patients, and 50 healthy subjects. All patients fulfilled the 1987 and 1982 classification criteria for RA and SLE, respectively, of the American College of Rheumatology. Demographic data and clinical characteristics of the patients were registered. Fasting lipid profile determination and leukocyte genomic DNA isolation from peripheral blood was performed in all the participants. Screening of the beta(3)-AR gene polymorphic region (exon 1) was done by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Quantitative and qualitative variables were analyzed using analysis of variance (ANOVA) with the LSD and chi(2) tests, respectively. An association between the arg64/arg64 beta(3)-AR genotype and high levels of triglycerides (TG) and very low-density lipoprotein cholesterol (VLDL-c) was found in three RA patients ( P=0.01), two of them taking chloroquine. Arg64/arg64 beta(3)-AR polymorphism may contribute to increased TG and VLDL-c in RA patients, independently of chloroquine treatment.
Assuntos
Antirreumáticos/uso terapêutico , Artrite Reumatoide/genética , Cloroquina/uso terapêutico , Lipídeos/sangue , Lúpus Eritematoso Sistêmico/genética , Receptores Adrenérgicos beta 3/genética , Adolescente , Adulto , Idoso , Artrite Reumatoide/sangue , Artrite Reumatoide/tratamento farmacológico , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/tratamento farmacológico , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de RestriçãoRESUMO
During the course of rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE), several immune and neuroendocrine changes associated with pregnancy may exert positive (amelioration) or negative (exacerbation) effects on the clinical outcome. In order to shed light on the mechanisms underlying these responses, we performed a prospective longitudinal study in RA and SLE pregnant women, including healthy pregnant women as a control group. Cytokine messenger RNA (mRNA) expression assessed by quantitative competitive polymerase chain reaction (PCR) in peripheral blood mononuclear cells (PBMC), cytokine levels and lymphocyte proliferation responses (LPR) following phytohaemagglutinin (PHA) stimulation of PBMC, plasma metalloprotease-9 activity (MMP-9) and hormonal status during pregnancy were determined. TNFa was the most abundant cytokine mRNA expressed in PBMC in all groups studied (healthy pregnant women, RA and SLE pregnant patients). However, a general TH2 response reflected by high IL-10 levels was found in RA, as well as SLE, patients. A significant change in IFN-gamma was observed in RA patients but only during the first trimester of pregnancy. This compared with a major TH1 response in healthy pregnant women. Interestingly, our study showed a homogeneous hormonal pattern in RA and SLE patients. Although decreased cortisol levels were observed in all patients studied, this is possibly related to the remission of disease activity status brought about by steroid treatment before and during pregnancy. In summary, we suggest that complex immune and hormonal networks are involved in pregnancy and that rheumatic diseases are very dynamic immune processes that cannot be described with a clear-cut cytokine profile. Furthermore, the observations in this study may reflect treatment-related immune effects more than those associated with disease.