RESUMEN
Plasmacytoid (p) dendritic cells (DC) are a specialized subset of DC whose primary role was initially defined by the production of type I interferons in response to viral infection. They are now known to also possess a repertoire of functions capable of determining T cell fate and activation. Under homeostatic conditions, non-lymphoid tissue-resident pDC play a critical role in the regulation of mucosal immunity, as well as the development of central and peripheral tolerance. Although these cells display a number of characteristics that differ from conventional DC, particularly altered costimulatory molecule expression and poor allostimulatory capacity when interacting with T cells, this phenotype favors the generation of alloantigen-specific regulatory CD4(+) or CD8(+) T cells critical to the development of graft tolerance. In this minireview, we discuss pDC ontogeny, functional biology and the emerging data that demonstrate the importance of pDC in the induction of tolerance, as well as recent studies that define mechanisms underlying pDC-mediated tolerance to both solid organ and haematopoietic stem cell transplants. We also highlight their use in clinical settings and the potential of pDC both as targets and cellular therapeutic agents to improve the outcome of organ transplantation.
Asunto(s)
Células Dendríticas/inmunología , Tolerancia Inmunológica , Inmunidad Mucosa/inmunología , Tolerancia al Trasplante , Trasplante de Células Madre Hematopoyéticas , HumanosRESUMEN
Thrombospondin-1 is a secreted protein that modulates vascular cell behavior via several cell surface receptors. In vitro, nanomolar concentrations of thrombospondin-1 are required to alter endothelial and vascular smooth muscle cell adhesion, proliferation, motility, and survival. Yet, much lower levels of thrombospondin-1 are clearly functional in vivo. This discrepancy was explained with the discovery that the potency of thrombospondin-1 increases more than 100-fold in the presence of physiological levels of nitric oxide (NO). Thrombospondin-1 binding to CD47 inhibits NO signaling by preventing cGMP synthesis and activation of its target cGMP-dependent protein kinase. This potent antagonism of NO signaling allows thrombospondin-1 to acutely constrict blood vessels, accelerate platelet aggregation, and if sustained, inhibit angiogenic responses. Acute antagonism of NO signaling by thrombospondin-1 is important for hemostasis but becomes detrimental for tissue survival of ischemic injuries. New therapeutic approaches targeting thrombospondin-1 or CD47 can improve recovery from ischemic injuries and overcome a deficit in NO-responsiveness in aging. (Part of a Multi-author Review).
Asunto(s)
Óxido Nítrico/metabolismo , Transducción de Señal , Trombospondina 1/fisiología , Animales , Humanos , Neovascularización FisiológicaRESUMEN
BACKGROUND AND PURPOSE: Angiogenesis involves multiple signaling pathways that must be considered when developing agents to modulate pathological angiogenesis. Because both cyclooxygenase inhibitors and dithioles have demonstrated anti-angiogenic properties, we investigated the activities of a new class of anti-inflammatory drugs containing dithiolethione moieties (S-NSAIDs) and S-valproate. EXPERIMENTAL APPROACH: Anti-angiogenic activities of S-NSAIDS, S-valproate, and the respective parent compounds were assessed using umbilical vein endothelial cells, muscle and tumor tissue explant angiogenesis assays, and developmental angiogenesis in Fli:EGFP transgenic zebrafish embryos. KEY RESULTS: Dithiolethione derivatives of diclofenac, valproate, and sulindac inhibited endothelial cell proliferation and induced Ser(78) phosphorylation of hsp27, a known molecular target of anti-angiogenic signaling. The parent drugs lacked this activity, but dithiolethiones were active at comparable concentrations. Although dithiolethiones can potentially release hydrogen sulphide, NaSH did not reproduce some activities of the S-NSAIDs, indicating that the dithioles regulate angiogenesis through mechanisms other than release of H(2)S. In contrast to the parent drugs, S-NSAIDs, S-valproate, NaSH, and dithiolethiones were potent inhibitors of angiogenic responses in muscle and HT29 tumor explants assessed by 3-dimensional collagen matrix assays. Dithiolethiones and valproic acid were also potent inhibitors of developmental angiogenesis in zebrafish embryos, but the S-NSAIDs, remarkably, lacked this activity. CONCLUSIONS AND IMPLICATION: S-NSAIDs and S-valproate have potent anti-angiogenic activities mediated by their dithiole moieties. The novel properties of S-NSAIDs and S-valproate to inhibit pathological versus developmental angiogenesis suggest that these agents may have a role in cancer treatment.
Asunto(s)
Anetol Tritiona/farmacología , Inhibidores de la Angiogénesis/farmacología , Antiinflamatorios no Esteroideos/farmacología , Ácido Valproico/farmacología , Animales , Proliferación Celular/efectos de los fármacos , Células Endoteliales/citología , Células Endoteliales/efectos de los fármacos , Proteínas de Choque Térmico HSP27 , Proteínas de Choque Térmico/metabolismo , Humanos , Ratones , Ratones Endogámicos C57BL , Fosforilación , Solubilidad , Pez CebraRESUMEN
Three-dimensional explant cultures of muscle tissue were used to characterize secreted proteins regulated by endogenous levels of the angiogenesis modulator thrombospondin (TSP)-1. Explants from TSP1 null mice exhibit enhanced neovascularization associated with increased endothelial outgrowth but decreased outgrowth of perivascular smooth muscle cells . The absence of endogenous TSP1 did not diminish activation of latent transforming growth factor-beta and moderately decreased matrix metalloproteinase levels. However, significant changes in other secreted proteins were observed. Endogenous TSP1 decreased mRNA levels for collagens Ialpha1, Ialpha2, and IIIalpha1 and laminin alpha4 and increased collagen IValpha1 mRNA expression. Endogenous TSP1 also decreased the level of type I collagen protein produced by the vascular outgrowths. Collagens Ialpha1, Ialpha2, and IIIalpha1 are known tumor endothelial markers, suggesting that TSP1 coordinately regulates a set of extracellular matrix genes that reverse the angiogenic switch. Suppression of collagen Ialpha1 or Ialpha2 mRNAs using antisense morpholinos inhibited outgrowth in TSP1 null explants and proliferation of TSP1 null endothelial cells, indicating that type I collagen synthesis is limiting for this neovascularization response.
Asunto(s)
Inhibidores de la Angiogénesis/fisiología , Colágeno Tipo I/genética , Regulación de la Expresión Génica , Trombospondina 1/fisiología , Animales , Movimiento Celular , Proliferación Celular , Células Cultivadas , Laminina/genética , Metaloproteinasa 2 de la Matriz/análisis , Metaloproteinasa 9 de la Matriz/análisis , Ratones , ARN Mensajero/análisis , Factor de Crecimiento Transformador beta/metabolismoRESUMEN
The blood-endothelial cell interface is a region of significant importance in many physiologic and pathologic processes. Blood-borne macromolecules and cells gain access to the subendothelial space and extravascular tissues by traversing the endothelium. Yet the various factors responsible for modulation of this process remain only partially elucidated. Several agents were found to be involved in this process, including nitric oxide (NO) and vascular endothelial growth factor (VEGF). It is known that under stress conditions (e.g., inflammation), NO can modulate the permeability of endothelial-cell monolayers to low-density mononuclear cells (LDMNCs). However, it is not known if NO can modulate such effects in the absence of inflammatory stimulation. In the present study, we utilized a Transwell chamber model to examine endothelial-cell monolayer permeability to LDMNCs in the absence of inflammatory stimuli. We noted that NO donor and L-arginine increased transendothelial-cell migration, whereas nitric oxide synthase (NOS) inhibition decreased migration. These effects were not significantly abrogated by VEGF antibody, suggesting that they were not VEGF-dependent.
Asunto(s)
Permeabilidad Capilar/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Células Endoteliales/efectos de los fármacos , Factores Relajantes Endotelio-Dependientes/farmacología , Óxido Nítrico/farmacología , Células Cultivadas , Humanos , Leucocitos Mononucleares/fisiología , Venas Umbilicales/efectos de los fármacosRESUMEN
Nine patients presented with non-healing venous ulcers of the lower limb. All had failed both non-surgical and surgical therapies. Following wide wound excision, perforator ligation, and microsurgical reconstruction, all wounds were healed. In two instances, separation at the flap/wound perimeter interface occurred and required additional dressing care to obtain wound closure. This was probably the result of incomplete excision of surrounding liposclerotic soft tissue. At 26 postoperative months, all wounds remained healed. Microsurgical transfer for properly selected patients can achieve healing of recalcitrant venous wounds, both over the intermediate and long term.
Asunto(s)
Colgajos Quirúrgicos , Úlcera Varicosa/cirugía , Anciano , Anastomosis Quirúrgica , Enfermedad Crónica , Femenino , Humanos , Masculino , Persona de Mediana Edad , Procedimientos de Cirugía Plástica , Resultado del Tratamiento , Cicatrización de HeridasRESUMEN
The tumor promotion stage of chemical carcinogenesis has been shown to exhibit a persistence of cellular effects during treatment and the reversibility of these changes upon cessation of treatment. Inhibition of gap-junctional intercellular communication and increased replicative DNA synthesis appear to be important in this process. The present study assessed the persistence and reversibility of gap-junctional intercellular communication inhibition, peroxisomal proliferation, and replicative DNA synthesis in livers from male F344 rats and B6C3F1 mice. Dietary administration of 20,000 mg/kg DEHP to male rats for 2 weeks decreased intercellular communication (67% of control) and enhanced replicative DNA synthesis (4.8-fold over control). Elevation of the relative liver weight and the induction of peroxisomal beta oxidation were also observed following treatment with 20,000 mg/Kg DEHP for 2 weeks. Following DEHP administration at a dose of 6000 mg/kg for 18 months, inhibition of gap-junctional intercellular communication persisted, and the relative liver weight and induction of peroxisomal beta oxidation remained elevated in both rats and male B6C3F1 mice. Treatment of rats and mice with phenobarbital for 18 months (500-mg/kg diet) also produced an increase in relative liver weight and a decrease in cell-to-cell communication. In recovery studies in which DEHP was administered to male F344 rats for 2 weeks and then withdrawn, the relative liver weight, rate of peroxisomal beta oxidation, increase in replicative DNA synthesis, and inhibition of gap-junctional intercellular communication returned to control values within 2 to 4 weeks after DEHP treatment ceased. Recovery studies with phenobarbital produced similar results. The primary active metabolite of DEHP, mono-2-ethylhexyl phthalate (MEHP), was detected in the livers of animals treated with DEHP for greater than 2 weeks. However, it could not be detected after removal of DEHP from the diet for 2 weeks. This study demonstrated that inhibition of gap-junctional intercellular communication, along with indicators of peroxisomal proliferation, including increased relative liver weight and enhanced peroxisomal beta oxidation, persist while DEHP treatment continues but reverses when treatment is stopped. Studies with phenobarbital produced a similar pattern of response.
Asunto(s)
Dietilhexil Ftalato/análogos & derivados , Dietilhexil Ftalato/farmacología , Hígado/efectos de los fármacos , Fenobarbital/farmacología , Animales , Peso Corporal/efectos de los fármacos , Comunicación Celular/efectos de los fármacos , Transformación Celular Neoplásica , Replicación del ADN/efectos de los fármacos , Dieta , Dietilhexil Ftalato/metabolismo , Ácidos Grasos/metabolismo , Uniones Comunicantes/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Masculino , Ratones , Ratones Endogámicos , Oxidación-Reducción , Peroxisomas/efectos de los fármacos , Peroxisomas/metabolismo , Ácidos Ftálicos/metabolismo , Ratas , Ratas Endogámicas F344RESUMEN
A number of chemical mediators can induce human keratinocytes and epidermal-derived carcinomas to undergo apoptosis, or programmed cell death. Recent evidence suggests pro-inflammatory cytokines, such as interleukin-1 beta or transforming growth factor alpha, protects carcinomas from numerous pro-apoptotic stimuli. Platelet-activating factor (1-alkyl-2-acetyl-3-glycerophosphocholine; PAF) is a lipid mediator with pro-inflammatory effects on numerous cell types. Although PAF can be metabolized to other bioactive lipids, the majority of PAF effects occur through activation of a G protein-coupled receptor. Using a model system created by retroviral transduction of the PAF receptor (PAF-R) into the PAF-R-negative human epidermal cell line KB and the PAF-R-expressing keratinocyte cell line HaCaT, we now demonstrate that activation of the epidermal PAF-R results in protection from apoptosis induced by tumor necrosis factor (TNF) alpha or TNF-related apoptosis-inducing ligand. The PAF-mediated protection was inhibited by PAF-R antagonists, and protection did not occur in PAF-R-negative KB cells. Additionally, we show protection from TNFalpha- or TRAIL-induced apoptosis by PAF-R activation is dependent on the transcription factor nuclear factor (NF)-kappa B, because PAF-R activation-induced NF-kappa B and epidermal cells transduced with a super-repressor form of inhibitor kappa B were not protected by the PAF-R. These studies provide a mechanism whereby the epidermal PAF-R, and possibly other G protein-coupled receptors, can exert anti-apoptotic effects through an NF-kappa B-dependent process.
Asunto(s)
Apoptosis/efectos de los fármacos , Epidermis/efectos de los fármacos , FN-kappa B/fisiología , Glicoproteínas de Membrana Plaquetaria/fisiología , Receptores de Superficie Celular , Receptores Acoplados a Proteínas G , Factor de Necrosis Tumoral alfa/farmacología , Línea Celular , Células Epidérmicas , Humanos , LigandosRESUMEN
The objective of this study was to assess the feasibility of digital patient satisfaction outcomes collection using the Internet and interactive voice response technology. Patients, health care providers, and industry sponsors were provided with incentives to participate. The participants included the practitioners and patients in medical and surgical practices who used Project Quality Card (patent pending) and the Practice Improvement Program. The study subjects were convenience samples of new and established patients seen between September 1999 and December 2000. There were 3 different pilot projects: QC1, QC2, and QC3. Patients were provided with a 20-minute prepaid phone card as an incentive for completing an interactive voice response call. Patients answered the 9-item visit rating questionnaire used to measure patient satisfaction. Patients responded on a 5-point scale, with 5 being excellent, and responses were totaled in the categories of patient access and physician attributes and in an overall score for the visit. A total of 998 patients from 77 physician offices participated. The activation responses, or percentage of patients using the Quality Cards, ranged from 12.8% in group 2 (QC2) to 26.6% in group 1 (QC1) to 34.8% in group 3 (QC3). This study demonstrates that incentivized digital outcomes collection can be successfully implemented in multisite community-based medical and surgical offices. The use of Project Quality Card (interactive voice response technology) and the Practice Improvement Program provides opportunities for user-friendly, benchmarked, real-time data collection. The percentage of patient activations is linked to provider participation in quality improvement programs. Patient participation is improved as the frequency of contact with a peer-directed database increases.
Asunto(s)
Encuestas de Atención de la Salud/métodos , Internet/estadística & datos numéricos , Motivación , Satisfacción del Paciente , Administración de la Práctica Médica/normas , Garantía de la Calidad de Atención de Salud/métodos , Recolección de Datos/métodos , Estudios de Factibilidad , Humanos , Medicina/normas , Proyectos Piloto , Indicadores de Calidad de la Atención de Salud , Especialización , Encuestas y Cuestionarios , Estados UnidosRESUMEN
The author presents the salvage of a long finger with osteomyelitis of the distal and middle phalanges, utilizing a vascularized hemi-metatarsal transfer.
Asunto(s)
Trasplante Óseo , Osteomielitis/cirugía , Infección de Heridas/cirugía , Adulto , Traumatismos de los Dedos/cirugía , Fracturas Óseas/cirugía , Humanos , Masculino , Metatarso/cirugía , Colgajos Quirúrgicos/irrigación sanguíneaRESUMEN
The author presents a case of osteomyelitis of the long bones, of over a half-century duration. An aggressive surgical approach comprising wide debridement of all compromised soft tissues and generous ostectomies, followed by myocutaneous flap reconstruction, achieved limb salvage.
Asunto(s)
Fémur , Osteomielitis/cirugía , Tibia , Anciano , Desbridamiento , Femenino , Humanos , Colgajos Quirúrgicos , Factores de TiempoRESUMEN
The present study evaluated the effect of di-2-ethylhexyl phthalate (DEHP) on gap-junctional intercellular communication (GJIC), peroxisomal beta-oxidation (PBOX) activity, and replicative DNA synthesis in several rodent species with differing susceptibilities to peroxisome proliferator-induced hepatic tumorigenesis. A low (non-tumorigenic) and high (tumorigenic) dietary concentration of DEHP was administered to male F344 rats for 1, 2, 4, and 6 weeks. Additionally, a previously non-tumorigenic dose (1000 ppm) and tumorigenic dose of DEHP (12,000 ppm), as determined by chronic bioassay data, were examined following 2 weeks dietary administration. Male B6C3F1 mice were fed the non-tumorigenic concentration, 500 ppm, and the tumorigenic concentration, 6000 ppm, of DEHP for two and four weeks. The hepatic effects of low and high concentrations of DEHP, 1000 and 6000 ppm, were also examined in male Syrian Golden hamsters (refractory to peroxisome proliferator-induced tumorigenicity). In rat and mouse liver, a concentration-dependent increase in the relative liver weight, PBOX activity, and replicative DNA synthesis was observed at the earliest time point examined. Concurrent to these observations was an inhibition of GJIC. In hamster liver, a slight increase in the relative liver weight, PBOX activity, and replicative DNA synthesis was observed. However, these effects were not of the same magnitude or consistency as those observed in rats or mice. Furthermore, DEHP had no effect on GJIC in hamster liver at any of the time points examined (2 and 4 weeks). HPLC analysis of DEHP and its primary metabolites, mono-2-ethylhexyl phthalate (MEHP), and phthalate acid (PA), indicated a time- and concentration-dependent increase in the hepatic concentration of MEHP. At equivalent dietary concentrations and time points, the presence of MEHP, the primary metabolite responsible for the hepatic effects of DEHP, demonstrated a species-specific response. The largest increase in the hepatic concentration of MEHP was observed in mice, which was greater than the concentration observed in rats. The hepatic concentration of MEHP was lowest in hamsters. Hepatic concentrations of DEHP and phthalic acid were minimal and did not correlate with concentration and time. Collectively, these data demonstrate the inhibition of hepatic GJIC and increased replicative DNA synthesis correlated with the observed dose- and species-specific tumorigenicity of DEHP and may be predictive indicators of the nongenotoxic carcinogenic potential of phthalate esters.
Asunto(s)
Comunicación Celular/efectos de los fármacos , Replicación del ADN/efectos de los fármacos , ADN/biosíntesis , Dietilhexil Ftalato/farmacología , Uniones Comunicantes/efectos de los fármacos , Hígado/efectos de los fármacos , Proliferadores de Peroxisomas/farmacología , Peroxisomas/efectos de los fármacos , Animales , Cromatografía Líquida de Alta Presión , Cricetinae , ADN/efectos de los fármacos , Dietilhexil Ftalato/análogos & derivados , Dietilhexil Ftalato/análisis , Dietilhexil Ftalato/metabolismo , Uniones Comunicantes/metabolismo , Hígado/metabolismo , Hígado/patología , Masculino , Mesocricetus , Ratones , Tamaño de los Órganos/efectos de los fármacos , Oxidación-Reducción , Peroxisomas/metabolismo , Ácidos Ftálicos/análisis , Ácidos Ftálicos/metabolismo , Ratas , Ratas Endogámicas F344 , Especificidad de la Especie , Aumento de Peso/efectos de los fármacosRESUMEN
The effects of the peroxisome proliferators di-isononyl phthalate (DINP) and di-2-ethylhexyl phthalate (DEHP) were evaluated in young adult male cynomolgus monkeys after 14 days of treatment, with emphasis on detecting hepatic and other effects seen in rats and mice after treatment with high doses of phthalates. Groups of 4 monkeys received DINP (500 mg/kg/day), DEHP (500 mg/kg/day), or vehicle (0.5% methyl cellulose, 10 ml/kg) by intragastric intubation for 14 consecutive days. Clofibrate (250 mg/kg/day), a hypolipidemic drug used for cholesterol reduction in human patients was used as a reference substance. None of the test substances had any effect on body weight or liver weights. Histopathological examination of tissues from these animals revealed no distinctive treatment-related effects in the liver, kidney, or testes. There were also no changes in any of the hepatic markers for peroxisomal proliferation, including peroxisomal beta-oxidation (PBOX) or replicative DNA synthesis. Additionally, in situ dye transfer studies using fresh liver slices revealed that DINP, DEHP, and clofibrate had no effect on gap junctional intercellular communication (GJIC). None of the test substances produced any toxicologically important changes in urinalysis, hematology, or clinical chemistry; however, clofibrate produced some emesis, small increases in serum triglyceride, decreased calcium, and decreased weights of testes/epididymides and thyroid/parathyroid. The toxicological significance of these small changes is questionable. The absence of observable hepatic effects in monkeys at doses that produce hepatic effects in rodents suggests that DINP, DEHP, and clofibrate would also not elicit in primates other effects such as liver cancer. These data, along with results from in vitro hepatocyte studies, indicate that rodents are not good animal models for predicting the hepatic effects of phthalates in primates, including humans.
Asunto(s)
Anticolesterolemiantes/toxicidad , Clofibrato/toxicidad , Dietilhexil Ftalato/toxicidad , Hígado/efectos de los fármacos , Macaca fascicularis , Peroxisomas/efectos de los fármacos , Ácidos Ftálicos/toxicidad , Administración Oral , Animales , Peso Corporal/efectos de los fármacos , Comunicación Celular/efectos de los fármacos , Replicación del ADN/efectos de los fármacos , Dietilhexil Ftalato/metabolismo , Uniones Comunicantes/efectos de los fármacos , Hígado/metabolismo , Hígado/patología , Tamaño de los Órganos/efectos de los fármacos , Oxidación-Reducción , Proliferadores de Peroxisomas/efectos adversos , Proliferadores de Peroxisomas/metabolismo , Peroxisomas/enzimología , Ácidos Ftálicos/metabolismoRESUMEN
The short-term hepatic effects of DINP (CAS 68515-48-0, designated DINP-1) in rats and mice were evaluated at tumorigenic and nontumorigenic doses from previous chronic studies. Groups of male F344 rats were fed diets with DINP-1 at concentrations of 0, 1000, or 12,000 ppm and male B6C3F1 mice at 0, 500, or 6000 ppm DINP-1. After 2 or 4 weeks of treatment, changes in liver weight, gap junctional intercellular communication (GJIC), peroxisomal beta-oxidation (PBOX), and replicative DNA synthesis were examined. In addition, hepatic and serum concentrations of the parent compound and major metabolites were determined. Relative to controls in both species, increased liver weight and PBOX at the high dose of DINP-1 were consistent with peroxisomal proliferation. Hepatic GJIC was inhibited and DNA synthesis was increased at the high dose of DINP-1, which is also consistent with the tumorigenic response in rats and mice reported in other chronic studies at these doses. These hepatic effects were not observed at the low doses of DINP-1. At comparable low doses of DINP-1 in other chronic studies, no liver tumors were observed in rats and mice. The monoester metabolite (MINP-1) was detected in the liver at greater concentrations in mice than rats. This result is also consistent with the dose-response observations in rat and mouse chronic studies. Additionally, other structurally similar dialkyl phthalate esters ranging from C7 to C11 were evaluated using a similar protocol for comparison to DINP-1; these included an alternative isomeric form of DINP (DINP-A), di-isodecyl phthalate (DIDP), di-isoheptyl phthalate (DIHP), di-heptyl, nonyl undecyl phthalate (D711P), and di-n-octyl phthalate (DNOP). Collectively, these data indicate that in rats and mice, DINP-1 and other C7-C11 phthalates exhibit a threshold for inducing hepatic cellular events. Further, where previous chronic data were available for these compounds, these phthalates elicited hepatic effects at doses that correlated with the tumorigenic response. Overall, these studies suggest a good correlation between the inhibition of GJIC when compared with the data on production of liver tumors in chronic studies.
Asunto(s)
Comunicación Celular/efectos de los fármacos , Replicación del ADN/efectos de los fármacos , ADN/biosíntesis , Uniones Comunicantes/efectos de los fármacos , Hígado/efectos de los fármacos , Peroxisomas/efectos de los fármacos , Ácidos Ftálicos/toxicidad , Animales , ADN/efectos de los fármacos , Uniones Comunicantes/metabolismo , Hígado/metabolismo , Masculino , Ratones , Ratones Endogámicos , Tamaño de los Órganos/efectos de los fármacos , Oxidación-Reducción , Peroxisomas/metabolismo , Ácidos Ftálicos/farmacocinética , Ratas , Ratas Endogámicas F344RESUMEN
Rotenone inhibits spontaneously and chemically induced hepatic tumorigenesis in rodents through the induction of apoptosis. However, the mechanism for the induction of apoptosis by rotenone has not been defined. Mitochondrial dysfunction, in particular the induction of the mitochondrial membrane permeability transition (MPT), has been implicated in the cascade of events involved in the induction of apoptosis. Inhibition of the mitochondrial electron-transport chain reduces the mitochondrial transmembrane potential (delta(psi)m), which may induce the formation of the mitochondrial permeability transition pore and the subsequent MPT. Fluorescent microscopy of Hoechst 33258-stained WB-F344 cells, a rat-liver cell line, was utilized to examine the effect of the mitochondrial respiratory chain inhibitor, rotenone (0.5-5 microM), atractyloside (5-10 microM), and cyclosporin A (2.5-10 microM) on apoptosis. A time- and concentration-dependent increase in liver cell apoptosis was observed following treatment with rotenone and atractyloside (11.7- and 7.7-fold, respectively, over solvent control). Cotreatment with 7.5- and 10 microM-cyclosporin A for 12 h inhibited the apoptogenicity of 5-microM rotenone treatment. A similar effect was observed following cyclosporin A cotreatment with atractyloside. Rotenone induced a rapid increase in apoptosis (within 20 min of treatment). By 2 h of treatment, the morphological appearance of apoptosis was similar to that observed in cultures treated continuously with rotenone for 12 h. Inhibition studies demonstrated that cyclosporin A prevented apoptosis if the exposure to it occurred prior to the 20-min threshold necessary to induce apoptosis by rotenone. Mitochondrial function was examined by staining with the mitochondrial membrane potential (delta(psi)m)-sensitive fluorochrome, MitoTracker Red (CMXRos) and confirmed utilizing cytofluorometric analysis of DiOC6(3)-stained cells. Rotenone (5.0-microM) and atractyloside (5.0-microM) reduced the percent of CMXRos or DiOC6(3)-positive (delta(psi)m-positive) liver cells within 15 min and throughout the duration of the study (6 h) to approximately 65-80% and 50-80% of control. However, cotreatment with concentrations of cyclosporin A that inhibited the apoptogenicity of rotenone and atractyloside prevented the rotenone- and atractyloside-induced reduction of the delta(psi)m. Therefore, the apoptogenic effect of rotenone and atractyloside appears to occur rapidly (within 20 min) and is irreversible once mitochondrial damage occurs. The inhibition of the rotenone- and atractyloside-induced apoptosis and mitochondrial dysfunction by cyclosporin A suggests the MPT may be involved in the induction of apoptosis by rotenone.
Asunto(s)
Apoptosis/efectos de los fármacos , Insecticidas/farmacología , Membranas Intracelulares/efectos de los fármacos , Hígado/efectos de los fármacos , Potenciales de la Membrana/efectos de los fármacos , Mitocondrias Hepáticas/efectos de los fármacos , Rotenona/farmacología , Animales , Atractilósido/farmacología , Línea Celular , Separación Celular , Supervivencia Celular/efectos de los fármacos , Ciclosporina/farmacología , Relación Dosis-Respuesta a Droga , Interacciones Farmacológicas , Citometría de Flujo , Membranas Intracelulares/fisiología , L-Lactato Deshidrogenasa/metabolismo , Hígado/citología , Hígado/fisiología , Potenciales de la Membrana/fisiología , Mitocondrias Hepáticas/fisiología , Permeabilidad/efectos de los fármacos , Ratas , Ratas Endogámicas F344 , Factores de TiempoRESUMEN
Reconstruction of the lower third of the leg and the forefoot remains a challenge due to a lack of regional muscle units and minimal subcutaneous tissues. Reverse island flaps have been applied to similar reconstructive problems in the upper extremity. Recently, the reverse sural artery neurocutaneous island flap has been utilized to reconstruct complex wounds of the lower extremity and forefoot in young and middle-aged individuals. We present our use of the flap in a patient cohort 65 years of age or older. Unique among this group was the high prevalence of diabetes and peripheral vascular disease. Nonetheless, the reverse sural artery neurocutaneous island flap proved a safe and reliable means of achieving wound closure.
Asunto(s)
Pie Diabético/cirugía , Úlcera de la Pierna/cirugía , Procedimientos de Cirugía Plástica/métodos , Colgajos Quirúrgicos , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Femenino , Humanos , Masculino , Presión , Úlcera por Presión/cirugía , Estudios RetrospectivosRESUMEN
A 36-year-old man sustained a thrombosis of the radial artery at the wrist while performing bench presses. Surgical exploration six weeks later demonstrated a 7-cm segmental thrombosis that was successfully bypassed with a reversed vein graft. The patient returned to full work duty without symptoms.
Asunto(s)
Arteria Radial/cirugía , Trombosis/etiología , Trombosis/cirugía , Levantamiento de Peso , Adulto , Humanos , Masculino , Muñeca/irrigación sanguíneaRESUMEN
A unique case of thumb salvage in a nine-year-old boy with degloving from a roping injury is presented. Immediate replantation was performed, but the part subsequently underwent arterial thrombosis and the soft-tissue unit was lost. Secondary resurfacing of the digit was achieved with a reverse island neurocutaneous radial artery island flap.
Asunto(s)
Amputación Traumática/cirugía , Arteria Radial/trasplante , Reimplantación/métodos , Terapia Recuperativa/métodos , Colgajos Quirúrgicos/irrigación sanguínea , Pulgar/cirugía , Amputación Traumática/etiología , Anastomosis Quirúrgica/métodos , Niño , Estudios de Seguimiento , Humanos , Masculino , Recuperación de la Función , Trasplante de Piel/métodos , Deportes , Pulgar/lesiones , Cicatrización de Heridas/fisiologíaRESUMEN
Six patients with massive post-extirpative wounds of the knee, all of whom had received adjuvant therapy, underwent wound closure and limb salvage via free-tissue transfer in all but one case. No instance of delayed wound healing requiring surgery occurred among the patients reconstructed by microsurgical tissue transfer. Five of the six patients are presently ambulating on the salvaged extremity. A protocol for management of these complex wounds is presented.