RESUMO
BACKGROUND: The aim of the study was to determine a rapid and simple method for isolation of polymorphonuclear (PMN) granulocytes from human blood appropriate for PCR of extracted DNA. METHODS: A modification of the combined dextran plus Ficoll-Hypaque methods was the final method used in 41 samples of normal white cells from humans. The changes encompassed a lower dextran-blood ratio (2:1), a shorter centrifugation time of leukocytes, a small volume of Ficoll-Hypaque suspension, and a decreased speed in the last centrifugation. The final key point was skipping the lysis of erythrocytes to avoid interference by salt solutions. RESULTS: These modifications rendered a mean PMN granulocyte recovery and purity of over 95% with a very narrow 2 - 8% coefficient of variation and did not alter the purity of DNA extraction. CONCLUSIONS: The combined dextran plus Ficoll-Hypaque method changes offer a fast and simple method to obtain an adequate purity of granulocytes for PCR.
Assuntos
Separação Celular , Granulócitos , Reação em Cadeia da Polimerase , Centrifugação com Gradiente de Concentração , Ficoll , HumanosRESUMO
BACKGROUND AND AIMS: In a previous uncontrolled experiment, oral vancomycin improved the symptoms (S) of chronic constipation (CC). The aim of this 21 day controlled pilot study was to determine if a low lincomycin dose improved the S of CC patients unresponsive to a high fiber diet. METHODS: On days 0-to-10, patients were randomized to 500 mg oral lincomycin + high fiber (L + F) or to placebo + high fiber (P + F). Participants and patients were blinded. From days 10-to-21, patients were continued solely on the high fiber diet. The primary efficacy endpoint was the difference in S between L + F and P + F from days 0-to-21 using a visual analog scale (VAS) calibrated from 0 = severe S to 10 = asymptomatic. RESULTS: The means of all S were significantly improved by L + F but not by P + F. A significant higher proportion of L + F patients increased the VAS > or = 3 points. CONCLUSIONS: The initial course of L facilitated the effect of F probably by its effect on the colon flora. This sequence of flora-altering biologics + F may serve as model to replace chronic use of drugs.
Assuntos
Antibacterianos/administração & dosagem , Constipação Intestinal/terapia , Fibras na Dieta/administração & dosagem , Lincomicina/administração & dosagem , Adulto , Idoso , Doença Crônica , Terapia Combinada , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND AND AIMS: In a previous uncontrolled experiment, oral vancomycin improved the symptoms (S) of chronic constipation (CC). The aim of this 21 day controlled pilot study was to determine if a low lincomycin dose improved the S of CC patients unresponsive to a high fiber diet. METHODS: On days 0-to-10, patients were randomized to 500 mg oral lincomycin + high fiber (L + F) or to placebo + high fiber (P + F). Participants and patients were blinded. From days 10-to-21, patients were continued solely on the high fiber diet. The primary efficacy endpoint was the difference in S between L + F and P + F from days 0-to-21 using a visual analog scale (VAS) calibrated from 0 = severe S to 10 = asymptomatic. RESULTS: The means of all S were significantly improved by L + F but not by P + F. A significant higher proportion of L + F patients increased the VAS > or = 3 points. CONCLUSIONS: The initial course of L facilitated the effect of F probably by its effect on the colon flora. This sequence of flora-altering biologics + F may serve as model to replace chronic use of drugs.
Assuntos
Antibacterianos/administração & dosagem , Constipação Intestinal/terapia , Fibras na Dieta/administração & dosagem , Lincomicina/administração & dosagem , Adulto , Adulto Jovem , Doença Crônica , Feminino , Humanos , Idoso , Masculino , Método Duplo-Cego , Pessoa de Meia-Idade , Projetos Piloto , Resultado do Tratamento , Terapia CombinadaRESUMO
BACKGROUND AND AIMS: In a previous uncontrolled experiment, oral vancomycin improved the symptoms (S) of chronic constipation (CC). The aim of this 21 day controlled pilot study was to determine if a low lincomycin dose improved the S of CC patients unresponsive to a high fiber diet. METHODS: On days 0-to-10, patients were randomized to 500 mg oral lincomycin + high fiber (L + F) or to placebo + high fiber (P + F). Participants and patients were blinded. From days 10-to-21, patients were continued solely on the high fiber diet. The primary efficacy endpoint was the difference in S between L + F and P + F from days 0-to-21 using a visual analog scale (VAS) calibrated from 0 = severe S to 10 = asymptomatic. RESULTS: The means of all S were significantly improved by L + F but not by P + F. A significant higher proportion of L + F patients increased the VAS > or = 3 points. CONCLUSIONS: The initial course of L facilitated the effect of F probably by its effect on the colon flora. This sequence of flora-altering biologics + F may serve as model to replace chronic use of drugs.