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1.
Leukemia ; 28(4): 813-22, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23989430

RESUMO

Prevailing wisdom holds that hematopoietic stem cells (HSCs) are predominantly quiescent. Although HSC cycle status has long been the subject of scrutiny, virtually all marrow stem cell research has been based on studies of highly purified HSCs. Here we explored the cell cycle status of marrow stem cells in un-separated whole bone marrow (WBM). We show that a large number of long-term multi-lineage engraftable stem cells within WBM are in S/G2/M phase. Using bromodeoxyuridine, we show rapid transit through the cell cycle of a previously defined relatively dormant purified stem cell, the long-term HSC (LT-HSC; Lineage(-)/c-kit(+)/Sca-1(+)/Flk-2(-)). Actively cycling marrow stem cells have continually changing phenotype with cell cycle transit, likely rendering them difficult to purify to homogeneity. Indeed, as WBM contains actively cycling stem cells, and highly purified stem cells engraft predominantly while quiescent, it follows that the population of cycling marrow stem cells within WBM are lost during purification. Our studies indicate that both the discarded lineage-positive and lineage-negative marrow cells in a stem cell separation contain cycling stem cells. We propose that future work should encompass this larger population of cycling stem cells that is poorly represented in current studies solely focused on purified stem cell populations.


Assuntos
Células da Medula Óssea/citologia , Ciclo Celular , Linhagem da Célula , Células-Tronco Hematopoéticas/citologia , Animais , Citometria de Fluxo , Masculino , Camundongos , Camundongos Endogâmicos C57BL
2.
J Neuroimmunol ; 184(1-2): 164-71, 2007 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-17275921

RESUMO

To test whether neutrophils (PMN) target lumbar dorsal root ganglia (DRG) following axonal injury leading to neuropathic pain, we visualized PMN infiltration in DRG tissue sections and estimated PMN count by flow cytometry following sciatic chronic constriction injury (CCI). Seven days after CCI, results show PMN within DRG where their count increased by three fold ipsilateral to injury compared to contralateral or sham, concomitant with peak neuropathic pain behavior. Superoxide burst in PMN isolated from rats d7 after CCI was elevated by 170% +/-18 compared to naïve and MCP-1 mRNA expression in DRG increased by 8.9+/-2.9 fold, but that of MIP-2, CINC-1, and RANTES did not change. We conclude that CCI causes PMN invasion of the DRG whereby the functional implication of their close proximity to neuronal axon and soma remains unknown.


Assuntos
Gânglios Espinais/patologia , Neutrófilos/fisiologia , Neuropatia Ciática/patologia , Neuropatia Ciática/fisiopatologia , Animais , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Constrição , Lateralidade Funcional , Regulação da Expressão Gênica/fisiologia , Região Lombossacral , Masculino , Medição da Dor/métodos , RNA Mensageiro/metabolismo , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo
3.
J Pediatr Surg ; 36(1): 220-8, 2001 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-11150470

RESUMO

BACKGROUND/PURPOSE: Biliary decompression for congenital or acquired obstruction (eg, biliary atresia) does not uniformly lead to liver repair, restore function, or prevent cholangitis. The authors hypothesize that failed repair is caused by altered macrophage (Mo) phenotypes central to an ongoing inflammatory and fibrogenic response. METHODS: In adult rats, biliary obstruction was performed by suspension of the common bile duct for 5 or 7 days. Decompression followed release of the loop until death during the course of liver repair. To determine Mo phenotype in the presence or absence of resident macrophages, animals were either administered gadolinium chloride or saline before injury and repair. At death, hepatic Mo were isolated, stained with MAC-1 (CD11b/CD 18) and OX-3 (MHC class II), and quantified with flow cytometry. Liver sections were immunostained for ED-1 and ED2; positive Mo were counted per square millimeter of tissue. RESULTS: Obstruction led to bile duct proliferation, fibrosis, and inflammation. Decompression relieved jaundice and ductal hyperplasia. After injury, hepatic Mo showed an 80% phenotypic conversion to MAC-1 and OX-3-positive cells. Cells isolated from livers at 9 days of repair persisted with 60% MAC-1 and 77% OX-3 expression. Gadolinium reduced Kupffer cells at all stages of repair. Recruited Mo in treated animals increased 4-fold greater than controls. CONCLUSIONS: Kupffer cells appear to limit the recruitment and persistence of a systemic macrophage phenotype in liver injury and repair. Cell surface markers for systemic macrophages appear after injury and persist during repair, despite adequate biliary decompression. After biliary decompression, this macrophage phenotype accounts for inflammatory complications such as cholangitis and ongoing fibrosis.


Assuntos
Colestase/imunologia , Colestase/cirurgia , Ativação de Macrófagos , Macrófagos/imunologia , Animais , Anti-Inflamatórios/farmacologia , Anticorpos Monoclonais , Bilirrubina/sangue , Colestase/patologia , Modelos Animais de Doenças , Citometria de Fluxo , Gadolínio/farmacologia , Técnicas Imunoenzimáticas , Células de Kupffer/fisiologia , Masculino , Fenótipo , Ratos , Ratos Sprague-Dawley
4.
J Surg Res ; 90(2): 166-73, 2000 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-10792959

RESUMO

BACKGROUND: During cholestatic liver injury, Kupffer cells (KC) and activated macrophages modulate cell proliferation and subsequent matrix deposition. The role of KC in the restoration of cell architecture and matrix metabolism during repair following chronic cholestatic liver injury is unknown. MATERIALS AND METHODS: To determine the effect of KC inactivation, adult male Sprague-Dawley rats underwent bile duct suspension (BDS) for 5 days followed by reversal of the obstruction. Saline (control) and gadolinium chloride (10 mg/kg) were administered 1 day prior to BDS and 1 day prior to reversal, to inactivate KC during both injury and repair. Serum bilirubin and quantitative cell morphometry were compared to verify the reversibility of the model. Collagen content of the liver was measured in trichrome-stained paraffin sections using NIH imaging software. RESULTS: Reversibility of the obstruction was verified by normalization of direct serum bilirubin, which peaked at 8.42 +/- 0.76 mg/dL following 5 days of BDS and returned to sham-operated levels 2 days after reversal, 0.36 +/- 0.15 mg/dL. Hematoxylin and eosin (H&E)-stained paraffin-embedded liver sections from gadolinium-treated animals at 4 and 7 days after reversal exhibited persistent bile duct proliferation, matrix deposition, and inflammation. Gadolinium-treated animals had altered collagen metabolism compared to saline controls. Whereas the collagen content in the saline group slowly returned to sham-operated levels over time, the treatment group demonstrated progressive accumulation of collagen during repair which was statistically significant at 7 days following reversal (8.79%/mm(2) +/- 2.17 in gadolinium group vs 2. 33%/mm(2) +/- 0.34 in saline group, P = 0.0003). CONCLUSIONS: These results demonstrate that inactivation of resident hepatic macrophages during liver repair impairs collagen metabolism, inhibits the resolution of fibrosis, and allows the persistence of inflammatory cell infiltrates in the portal areas. This is the first evidence of profibrogenic responses in the absence of an intact KC compartment during repair after cholestatic injury.


Assuntos
Colestase Extra-Hepática/fisiopatologia , Células de Kupffer/fisiologia , Regeneração Hepática/fisiologia , Animais , Ductos Biliares/patologia , Ductos Biliares/cirurgia , Bilirrubina/sangue , Peso Corporal , Doença Hepática Induzida por Substâncias e Drogas , Colestase Extra-Hepática/patologia , Colestase Extra-Hepática/cirurgia , Doença Crônica , Colágeno/análise , Colágeno/metabolismo , Meios de Contraste , Modelos Animais de Doenças , Células Epiteliais/química , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Fibrose , Gadolínio , Hiperplasia , Células de Kupffer/metabolismo , Células de Kupffer/patologia , Hepatopatias/patologia , Masculino , Ratos , Ratos Sprague-Dawley
5.
J Pediatr Gastroenterol Nutr ; 19(2): 164-9, 1994 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-7815237

RESUMO

Although population-based studies have shown that children have the highest age-specific incidence of infection with the Shiga-like toxin-producing E. coli (SLTEC), these sporadic case series were not focused specifically on the pediatric age group. We undertook a prospective study to determine the frequency of detection of SLT in an exclusively pediatric population. The study design minimized ascertainment and referral bias by systematically defining the population by the presence of diarrheal symptoms rather than by specific diagnosis, previous submission of stool for culture, or referral to a diarrhea study. All children < 10 years of age hospitalized at a tertiary care pediatric hospital, irrespective of admission diagnosis, were surveyed prospectively at admission and for 2 days thereafter for the presence of defined diarrheal symptoms. From May 1, 1991, to April 30, 1992, 227 patients and 92 age- and season-matched controls were enrolled. Fecal SLT was detected in six (2.6%) patients, three of whom had E. coli O157:H7 organisms were isolated; SLT was not found in any of the controls. SLT was more commonly detected in children 2-10 years of age and in bloody stools. Salmonella was isolated in six (2.6%) cases, Shigella in five (2.2%), and Yersinia in three (1.3%); rotavirus was detected in 46 (20.3%). Two patients with SLT-associated diarrhea had hemolytic uremic syndrome (HUS), and four had hemorrhagic colitis. SLT-associated diarrhea occurred in the summer and fall months in contradistinction to that with rotavirus, which occurred in the winter and spring.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Toxinas Bacterianas/análise , Diarreia/microbiologia , Infecções por Escherichia coli/microbiologia , Escherichia coli/isolamento & purificação , Toxinas Bacterianas/biossíntese , Criança , Pré-Escolar , Escherichia coli/metabolismo , Fezes/química , Fezes/microbiologia , Humanos , Lactente , Recém-Nascido , Estudos Prospectivos , Toxina Shiga I , Toxina Shiga II
6.
J Antimicrob Chemother ; 25(1): 73-81, 1990 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-2156792

RESUMO

Two unusual, heterogeneously-resistant, strains of Staphylococcus aureus appeared resistant to oxacillin, but susceptible to methicillin by disc diffusion methods. In agar dilution tests, both strains were oxacillin-resistant. One was susceptible to methicillin, and the other gave a paradoxical reaction, with growth only on plates containing low (0.5, 1 and 2 mg/l) and high (32 and 64 mg/l) concentrations of antibiotic. Induction of methicillin resistance was tested by inoculating each strain on to agar plates containing an inhibitory concentration of methicillin (8 mg/l), and then placing discs containing inducers (oxacillin, nafcillin, methicillin and CBAP [2-(2'-carboxyphenyl) benzoyl-6-aminopenicillanic acid]) on the agar surface. Colonies grew only around discs containing effective inducers. Oxacillin and CBAP were much more potent inducers of methicillin resistance and beta-lactamase than was nafcillin or methicillin. These data suggest that the mechanism that regulates induction of the low-affinity penicillin binding protein (PBP-2') may be altered in these strains. Similar mechanisms appear to induce both beta-lactamase and methicillin resistance.


Assuntos
Proteínas de Bactérias , Hexosiltransferases , Meticilina/farmacologia , Peptidil Transferases , beta-Lactamases/metabolismo , Proteínas de Transporte/biossíntese , Divisão Celular/efeitos dos fármacos , Ácidos Clavulânicos/farmacologia , Resistência Microbiana a Medicamentos , Indução Enzimática , Testes de Sensibilidade Microbiana , Muramilpentapeptídeo Carboxipeptidase/biossíntese , Nafcilina/farmacologia , Oxacilina/farmacologia , Ácido Penicilânico/farmacologia , Resistência às Penicilinas , Proteínas de Ligação às Penicilinas , Staphylococcus aureus/genética
7.
J Clin Microbiol ; 26(7): 1425-7, 1988 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-3410957

RESUMO

Clinical isolates of Staphylococcus aureus wee inappropriately categorized as intermediate or resistant to oxacillin on the basis of tests with two lots of oxacillin disks. The potency of one lot was tested and found to be below accepted limits. Routine quality control tests failed to detect the defective disks.


Assuntos
Testes de Sensibilidade Microbiana/normas , Oxacilina/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Humanos , Resistência às Penicilinas , Controle de Qualidade
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