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@#Parthanatos is a form of programmed cell death,which is also known as poly(ADP-ribose)polymerase 1(PARP1)-mediated apoptosis-inducing factor(AIF)and macrophage migration inhibitory factor(MIF)-dependent cell death according to its molecular mechanism. Parthanatos is the main cause of a variety of neurodegenerative diseases,such as Parkinson's disease(PD),Alzheimer's disease(AD),motor neuron disease,and is also involved in the pathogenesis of some tumors,such as lung cancer and breast cancer. Therefore,a thorough understanding of the molecular mechanism of Parthanatos is crucial for the therapeutic strategies of related diseases. In recent years,studies have found that effective regulation of the occurrence of Parthanatos by regulating the key proteins PARP1,AIF and MIF is expected to become a therapeutic target for many diseases. Based on the specific molecular mechanism of Parthanatos,this paper reviewed the research progress of therapeutic strategies for related diseases from the aspects of inhibiting and promoting Parthanatos.
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Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.T260A, p.R422W, and p.R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‒49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‒17.9%, which was significantly higher than that (6.9%‒7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.
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Humans , Apoptosis Inducing Factor/metabolism , NAD/metabolism , Dimerization , ApoptosisABSTRACT
Objective:To investigate the effects of naringin on early brain injury in rats with subarachnoid hemorrhage and its possible mechanism of action.Methods:Rats were randomly divided into the sham operation group, the model group, and the naringin group. Each group had 8 rats. The SAH model was established by intravascular perforation, and then rats in the model group and the naringin group were administered 0.9% NaCl or naringin 40 mg/kg by intraperitoneal injection after 0.5 h. SAH score, neurological function score, cerebral edema, and blood-brain barrier permeability were detected. The level of NAD + and nflammatory factors were detected by ELISA. The expression of poly(ADP-ribose) polymerase-1 (PARP-1), apoptosis inducing factor (AIF), and protease-activated receptor (PAR) proteins was detected by Western Blot. The expression of PARP-1 mRNA was detected by quantitative real-time fluorescence PCR (qRT-PCR). Neuronal apoptosis was detected by an immunofluorescence assay. Results:Compared with the model group, naringin treatment improved neurological function ( P<0.01), reduced cerebral edema and Evans blue exudation (all P<0.01), increased the content of NAD + ( P<0.001), reduced IL-1β, IL-6 and TNF-α levels (all P<0.001), and reduced the expression of PARP-1/AIF pathway-related proteins in vivo (all P<0.001). In addition, naringin could inhibit neuronal apoptosis in early brain injury after SAH. Conclusions:Naringin can improve the early brain injury after SAH, which may be achieved by inhibiting the PARP-1/AIF pathway.
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Acetaminophen (APAP) is a widely used analgesic and antipyretic drug, which is safe at therapeutic doses but can cause severe liver injury and even liver failure after overdoses. The mouse model of APAP hepatotoxicity recapitulates closely the human pathophysiology. As a result, this clinically relevant model is frequently used to study mechanisms of drug-induced liver injury and even more so to test potential therapeutic interventions. However, the complexity of the model requires a thorough understanding of the pathophysiology to obtain valid results and mechanistic information that is translatable to the clinic. However, many studies using this model are flawed, which jeopardizes the scientific and clinical relevance. The purpose of this review is to provide a framework of the model where mechanistically sound and clinically relevant data can be obtained. The discussion provides insight into the injury mechanisms and how to study it including the critical roles of drug metabolism, mitochondrial dysfunction, necrotic cell death, autophagy and the sterile inflammatory response. In addition, the most frequently made mistakes when using this model are discussed. Thus, considering these recommendations when studying APAP hepatotoxicity will facilitate the discovery of more clinically relevant interventions.
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The chronic consumption of alcohol causes a worsening of the events that follow the cerebral ischemia. These events are regulated through the expression of several genes and microRNAs. The aimof this work was To analyze and describe the expression profile of PARP and AIF and miRNA-9 proteins in rats submitted to focal cerebral ischemia, associated or not with chronic alcoholism model. Methods: Twenty adult Wistar rats, subdivided into: control; ischemic; alcoholic and ischemic / alcoholized for immunohistochemical analysis and miRNA-9 gene expression. Results: There was a reduction in the protein expression of PARP-1 and a positive marking for AIF in the ischemic / alcoholized group. The miRNA-9 did not obtain significant expression. The association of ischemia with chronic alcohol use promoted a tendency to low expression of miRNA-9, low expression of PARP-1 and high expression of AIF, indicating an interference in the protective effect of miRNA-9 be observed in the other groups.
El consumo crónico de alcohol provoca un empeoramiento de los eventos que siguen a la isquemia cerebral. Estos eventos están regulados a través de la expresión de varios genes y microRNA. El objetivo de este trabajo fue analizar y describir el perfil de expresión de las proteínas PARP y AIF y microRNA-9 en ratas sometidas a isquemia cerebral focal, asociadas o no, con el modelo de alcoholismo crónico. Veinte ratas Wistar adultas se dividieron en: grupo control, isquémico alcohólico, e isquémico / alcoholizado para análisis inmunohistoquímico y expresión de genes microRNA-9. Resultados: Hubo una reducción en la expresión de proteínas de PARP-1 y un marcado positivo para AIF en el grupo isquémico / alcoholizado. No se observó una expresión significativa en el microRNA-9. La asociación de la isquemia con el consumo crónico de alcohol promovió una tendencia a la baja expresión de microRNA-9, baja expresión de PARP1 y alta expresión de AIF, lo que indica una interferencia en el efecto protector de microRNA-9 en los otros grupos.
Subject(s)
Animals , Rats , Brain Ischemia/metabolism , Alcoholism/metabolism , Immunohistochemistry , Brain Ischemia/genetics , Rats, Wistar , MicroRNAs/metabolism , Disease Models, Animal , Alcoholism/genetics , Apoptosis Inducing Factor/metabolism , Poly (ADP-Ribose) Polymerase-1/metabolismABSTRACT
Solute carrier (SLC) transporters meditate many essential physiological functions, including nutrient uptake, ion influx/efflux, and waste disposal. In its protective role against tumors and infections, the mammalian immune system coordinates complex signals to support the proliferation, differentiation, and effector function of individual cell subsets. Recent research in this area has yielded surprising findings on the roles of solute carrier transporters, which were discovered to regulate lymphocyte signaling and control their differentiation, function, and fate by modulating diverse metabolic pathways and balanced levels of different metabolites. In this review, we present current information mainly on glucose transporters, amino-acid transporters, and metal ion transporters, which are critically important for mediating immune cell homeostasis in many different pathological conditions.
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Objective To investigate the effect of thyroid hormone(T3)on the expressions of cytochrome c (CytC) and apoptosis?inducing factor(AIF) after cerebral ischemia reperfusion injury in rats and its mechanism. Methods SD male rats were randomly divided into four groups: sham operation group(sham1),sham operation group + T3(sham2) ,ischemia?reperfusion group (IR) ,and thyroid hormone treatment group(T3). A rat model of cerebral ischemia?reperfusion injury was established by right middle cerebral artery occlusion for 2 h,followed by reperfusion for 24 h. Thyroid hormones (10μg/100 g) or normal saline were given at 1 h after onset of ischemia and 6 h after reperfusionby intraperitoneal injection. Neurological deficit scores were evaluated 24 h after reperfusion. Cerebral infarction volume was evaluated by TTC staining. Histological changes was observed by HE staining. Expressions and mRNA levels of CytC and AIF in ischemic brain tissue were evaluated by immunohistochemistry and real?time fluorescent quantitative RT?PCR. Results Ascompared with those in sham operation groups ,the expressions and mRNA levels of CytC and AIFincreased significantlyin IR groups. As compared with those in IR groups ,the indexes were remarkably decreasedin T3 groups (P < 0.01). Nerve function was markedly improved andinfarction area narrowed.Conclusions Thyroid hormone plays a certain role in protection of cerebral ischemia?reperfusion injury in rats,whose mechanism may be associated with inhibition of the expression of apoptosis factors?CytC and AIF.
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Objective To explore the effect of Quercetin on the long-term memory and PARP-1/AIF signal path in neonatal rats with hypoxic-ischemic brain damage (HIBD). Methods Fifty-six 7-day-old SD rats were randomly divided into sham-operation group, HIBD group, low dose of Quercetin group (20 mg/kg), and high dose of Quercetin group (40 mg/kg), each of 14 rats. Except for sham-operation group, in the other groups HIBD model were made by right common carotid artery ligation and anoxiate. The Quercetin groups were injected with the corresponding doses of Quercetin immediately once a day continuously for 7 days after the model was made,. Sham-operation group and HIBD group were intraperitoneally injected with normal saline at the same time. Neural function was evaluated by Hanging wire test and Vertical pole test at 21 days old. The capacity of learning and memory was detected by Morris water maze at 28 days old, and then rats were killed and brains were taken. HE was used to observe the pathological changes of hippocampus. Western blot were used to detect the expression of PARP-1 and AIF in hippocampus. Results Compared with sham-operation group, the neural function and learning and memory ability decreased significantly in HIBD group. Those ability in both low dose and high dose of Quercetin groups were remarkably increased in comparison with HIBD group, and there were statistic differences (P??0.05). Conclusion Quercetin could improve long-term learning memory in newborn rats with HIBD, and the mechanism may be down-regulation of PARP-1/AIF cell apoptosis signaling pathway, inhibition of neuronal apoptosis, and thus play a role in protection of brain.
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Aim To observe histopathological changes of hippocampus after acute epilepsy induced by penty-lenetetrazole (PTZ)in rats.Methods Five groups as control group,PTZ-induced 24 hours(h)group,PTZ-induced 72 hours group,PTZ-induced 1 20 hours group and PTZ-induced 1 44 hours group were designed.PTZ (64 mg·kg -1 )was administered with a single intrap-eritoneal injection for generalized tonic-clonic sei-zures in the current experiment.Control and PTZ trea-ted animals were sacrificed after specific time points. Brain was dissected out and then evaluated for neuro-pathological changes using Nissl staining and immuno-histochemical technique.Results In this study PTZ-induced hippocampal neuron status apoptosis occurred at 24 hours and was sustained for 1 44 hours after status epilepticus.Whereas,activated caspase-3 and AIF ap-peared at 24 hours and were sustained for 1 44 hours af-ter status epilepticus.Conclusion The results of this study show that the significant histopathological chan-ges of hippocampus appear in the vicinity of 1 20 hours after intraperitoneal injection of pentylenetetrazole.
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Ischemic stroke and ischemia/reperfusion (I/R) injury induced by thrombolytic therapy are conditions with high mortality and serious long-term physical and cognitive disabilities. They have a major impact on global public health. These disorders are associated with multiple insults to the cerebral microcirculation, including reactive oxygen species (ROS) overproduction, leukocyte adhesion and infiltration, brain blood barrier (BBB) disruption, and capillary hypoperfusion, ultimately resulting in tissue edema, hemorrhage, brain injury and delayed neuron damage. Traditional Chinese medicine (TCM) has been used in China, Korea, Japan and other Asian countries for treatment of a wide range of diseases. In China, the usage of compound TCM preparation to treat cerebrovascular diseases dates back to the Han Dynasty. Even thousands of years earlier, the medical formulary recorded many classical prescriptions for treating cerebral I/R-related diseases. This review summarizes current information and underlying mechanisms regarding the ameliorating effects of compound TCM preparation, Chinese materia medica, and active components on I/R-induced cerebral microcirculatory disturbances, brain injury and neuron damage.
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Objective The study aimed to explore the relationship between AIF related pathway and the inju-ring of cultured SGNs (spiral ganglion neurons)by glutamate toxicity,and to find AIF expression and distribution changes in SGNs.Methods SGNs of 40 newborn rats within 3 day were obtained and cultured in vitro.Cultured cells were divided into four groups:the normal control group,10 mM,20 mM and 40 mM glutamate injured group, separately.After 48 h hours culturing,optical microscopy,immune fluorescence staining and real-time fluores-cence quantitative PCR were used to observe the morphology,AIF distribution,and AIF,calpain,Caspase3 expres-sion changes in SGNs in vitro.TUNEL was used to verify the cell apoptosis.ResuIts Noticeable morphological chan-ges and cell apoptosis were occurred in 20 mM glutamate group,with AIF nuclear translocation.AIF gene expression was significantly higher than normal after glutamate administration (P0.05). ConcIusion In the process of cultured SGNs injured by glutamate,AIF participated in the cell apoptosis.Noticeable cell apoptosis were occurred in 20 mM glutamate group with AIF nuclear translocation.Calpain up-expression also contributed to excitatory neurotransmitter injury on SGNs,but Caspase 3 had no obvious effects.
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Apoptosis-inducing factor (AIF) is a mitochondrial flavoprotein that mediates both NADH-oxidizing and caspase-independent apoptosis. Further, the proapoptotic activity of AIF is located in the C-terminus of AIF, although the precise minimum sequence responsible for apoptosis induction remains to be investigated. In the present study, we generated two truncated AIFs, AIFΔ1-480-FLAG, which is a FLAG-tagged C-terminal peptide comprising amino acids from 481 to 613, and AIF360-480 containing amino acids from 360 to 480 of AIF. We used confocal microscopy to demonstrate that both the truncated proteins are expressed and located in the cytoplasm of transfected cells. AIFΔ1-480 but not AIF360-480 induces apoptosis in transfected cells. We also found that the expression of AIFΔ1-480 could initiate the release of cytochrome c from the mitochondria. The suppression of caspase-9 via siRNA blocked the proapoptotic activity of AIFΔ1-480. Therefore, AIFΔ 1-480 is sufficient for inducing caspase-9-dependent apoptotic signaling, probably by promoting the release of cytochrome c. At last, we generated a chimeric immuno-AIFΔ 1-480 protein, which comprised an HER2 antibody, a Pseudomonas exotoxin A translocation domain and AIFΔ 1-480. Human Jurkat cells transfected with the immuno-AIFΔl-480 gene could express and secrete the chimeric protein, which selectively recognize and kill HER2-overexpressing tumor cells. Our study demonstrates the feasibility of the immuno-AIFΔl-480 gene as a novel approach to treating HER2-overexpressing cancers.
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Humans , Alcohol Oxidoreductases/drug effects , Apoptosis Inducing Factor/pharmacology , Apoptosis/drug effects , DNA, Complementary/drug effects , DNA-Binding Proteins/drug effects , Alcohol Oxidoreductases/genetics , Alcohol Oxidoreductases/metabolism , Apoptosis Inducing Factor/genetics , Apoptosis Inducing Factor/metabolism , Apoptosis/genetics , Blotting, Western , DNA, Complementary/genetics , DNA, Complementary/metabolism , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Fluorescent Antibody Technique, Indirect , Jurkat Cells , Microscopy, Confocal , Reverse Transcriptase Polymerase Chain Reaction , TransfectionABSTRACT
Objective To preliminary investigate the cytoactive of HELA cell after PUMA gene promoting HELA cell apoptosis and whether or not the protein of cyt-c,AIF in the chondrosome have participated in the process of PUMA gene inducing HELA cell.Methods we Used AO/EB dyeing to detect the Morphologic change of cell induces by PUMA gene and Western Blot to detect whether or not the position of cyt-c,AIF in the chondrosome have transferred after PUMA gene induces HELA cell apoptosis.Result ①Fluorescence microscope and inverted phase contrast microscope display HELA cell have a series of apoptosis Morphologic change after transfecting PUMA into HELA cell,and this Morphologic change of cell is more obviously after transfecting 48h than after 24h.;②Western Blot display the protein cyt-c and AIF transferred toward to kytoplasm after HELE cell took place apoptosis,the mount of transfecting protein is more obviously after transfecting 48h than after 24h.Conclusion ① PUMA gene have the effect of promoting HELA cell apoptosis;②cyt-c and AIF take part in the process of promoting HELA cell apoptosis,but during this process if the membrane potential of chondrosome was changing need to do some more study to confirm it.
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The caudate-putamen is generally referred to as the striatum or neostriatum, which is one of the main components of the basal ganglia and this designation, is especially relevant in the rat. In spite of the fact that it comprises one of the major parts in central nervous system, studies on how seriously vulnerable this area to cerebral ischemia especially in neonates is still remained. In this study, using neonatal (7 days postnatal) rats, we speculated the significance of AIF in the apoptotic neuronal cell death in basal ganglia induced by hypoxic-ischemic injury. For this study, we introduced permanent common carotid artery occlusion and then, exposed to 6% oxygen for 2 hours and the results are as follows: 1. There were tendency of increasing AIF immunoreactivity induced by hypoxic-ischemic insult in neonatal basal ganglia at 12 & 24 hrs after hypoxic-ischemic insult. 2. Western Blotting analysis showed increased AIF expression in basal ganglia at 12 hrs(caudate-putamen) & 24 hrs (globus pallidus) after hypoxic-ischemic insult. 3. Evidence of localization of AIF in glial cells as well as in neurons obtained by double-immunofluorescence staining. Our results seem to provide evidences on the involvement of AIF in the apoptotic neuronal cell loss with hypoxicischemic insult in neonatal rat. Furthermore, localization AIF in glial cells as well as in neurons suggests involvement of neuroglial cells in the apoptotic pathway in neonatal basal ganglia induced by hypoxic-ischemic injury.
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Animals , Humans , Infant, Newborn , Rats , Basal Ganglia , Blotting, Western , Brain Ischemia , Brain , Carotid Artery, Common , Cell Death , Central Nervous System , Neostriatum , Neuroglia , Neurons , OxygenABSTRACT
delta12-Prostaglandin (PG) J2 is known to elicit an anti-neoplastic effects via apoptosis induction. Previous study showed delta12-PGJ2-induced apoptosis utilized caspase cascade through cytochrome c-dependent pathways in HeLa cells. In this study, the cellular mechanism of delta12-PGJ2- induced apoptosis in HeLa cells, specifically, the role of two mitochondrial factors; bcl-2 and apoptosis-inducing factor (AIF) was investigated. Bcl-2 attenuated delta12-PGJ2-induced caspase activation, loss of mitochondrial transmembrane potential (delta psi m), nuclear fragmentation, DNA laddering, and growth curve inhibition for approximately 24 h, but not for longer time. AIF was not released from mitochondria, even if the delta psi m was dissipated. One of the earliest events observed in delta12-PGJ2-induced apoptotic events was dissipation of delta psi m, the process known to be inhibited by bcl-2. Pre-treatment of z-VAD- fmk, the pan-caspase inhibitor, resulted in the attenuation of delta psi m depolarization in delta12-PGJ2- induced apoptosis. Up-regulation of Sox-4 protein by delta12-PGJ2 was observed in HeLa and bcl-2 overexpressing HeLa B4 cell lines. Bcl-2 overexpression did not attenuate the expression of Sox-4 and its expression coincided with other apoptotic events. These results suggest that delta12-PGJ2 induced Sox-4 expression may activate another upstream caspases excluding the caspase 9-caspase 3 cascade of mitochondrial pathway. These and previous findings together suggest that delta12-PGJ2-induced apoptosis in HeLa cells is caspase-dependent, AIF-independent events which may be affected by Sox-4 protein expression up-regulated by delta12-PGJ2.
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Female , Humans , Amino Acid Chloromethyl Ketones/pharmacology , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Caspases/physiology , Cytochromes c/physiology , Flavoproteins/metabolism , HeLa Cells , High Mobility Group Proteins/physiology , Membrane Proteins/metabolism , Mitochondria/metabolism , Prostaglandin D2/pharmacology , Protein Transport/physiology , Proto-Oncogene Proteins c-bcl-2/biosynthesis , Transcriptional Activation , Trans-Activators/physiologyABSTRACT
PURPOSE: This study was undertaken to reveal the molecular mechanism underlying sulindac-induced apoptosis in the human colon cancer cell line HT-29 (mutant p53). METHODS: Apoptosis was determined by using Hoechst 33342 staining, and translocation of proteins was established by using immunofluorescence, immunoelectron microscopy, and Western blotting after ultra- fractionation. RESULTS: This type of apoptosis was associated with decreased mitochondrial membrane potential, a translocation of the apoptosis-inducing factor (AIF) to the nucleus, and morphological evidence of nuclear condensation. However, DNA electrophoresis did not elucidate the ladder pattern of DNA fragments. Instead, a pulse-field gel electrophoresis showed that sulindac led to disintegration of nuclear DNA into-high- molecular-weight DNA fragments of about 100~300 kbp. CONCLUSIONS: Our findings indicate that sulindac induces large-scale DNA fragmentation, suggesting a predominantly AIF-mediated cell-death process, through translocation of the AIF to the nucleus in HT-29 cells.
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Humans , Apoptosis Inducing Factor , Apoptosis , Blotting, Western , Cell Line , Colonic Neoplasms , DNA Fragmentation , DNA , Electrophoresis , Fluorescent Antibody Technique , HT29 Cells , Membrane Potential, Mitochondrial , Microscopy, Immunoelectron , SulindacABSTRACT
It is well known that cerebral ischemia induces apoptotic neuronal cell death along with necrosis. Apoptosis is characterized by several morphological nuclear changes including chromatin condensation and nuclear fragmentation. These changes are triggered by the activation of members of caspase family, caspase activated DNase, and several novel proteins. A novel gene, the product of which caused chromatin condensation and DNA fragmentation, was recently identified, cloned, and designated apoptosis inducing factor (AIF). AIF is a mitochondrial flavoprotein (prosthetic group: flavin adenine dinucleotide) with significant homology to plant ascorbate reductase and bacterial NADH oxidase, and is normally confined to the mitochondrial space. In a variety of different apoptosis-inducing conditions, AIF translocates through the outer mitochondrial membrane to the cytosol and the nucleus. Thus, similar to cytochrome c, AIF is a phylogenetically old, bifunctional protein with an oxidoreductase function and a second apoptogenic function. In this study, to investigate changes of AIF expression levels in the rat brain during various time periods (6, 12, 18, 24 hrs) of ischemic insults, we performed permanent middle cerebral artery (MCA) occlusion model using male SD rats and the extent of ischemic injury was measured by 2% TTC (2, 3, 5-triphenyl tetrazolium chloride) staining. There were numerous TUNEL-positive neuronal cell nuclei, which implies apoptotic cell death in the areas with ischemic insult. Western blot analysis was performed with separate samples (cytosolic fraction & mitochondrial fraction). This method was designed to investigate if the expression of AIF is really increased during ischemic injury, or the increased AIF activity is caused by its release from the mitochondrial inter-membrane space to cytosol. In the immunohistochemical study using polyclonal anti-AIF antibody revealed overall distribution of AIF in the rat brain including cerebral cortex, basal ganglia and also in hippocampus that is especially sensitive to ischemia. There was apparently increased AIF immunoreactivity in the nucleus accumbens of ischemia-induced hemisphere compared to contralateral control hemisphere. Our results seem to be the first evidence on the involvement of AIF in the apoptotic neuronal cell loss with ischemic insult and will provide some clues to the pathophysiology of neuronal apoptosis induced by ischemic insult. Further researches will reveal functional relationship between AIF and other factors involved in chromatin condensation and degradation and whether AIF itself will be useful as a caspase-independent, Bcl-2-independent death inducer.
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Male , Humans , Rats , AnimalsABSTRACT
STUDY DESIGN: We have analyzed the laparoscopic lateral retroperitoneal approach of the L4-5 interspace to the miniopen retroperitoneal approach for lateral lumbar interbody fusion. OBJECTIVES: To prospectively compare the laparoscopic lateral retroperitoneal approach of the L4-5 interspace to the miniopen retroperitoneal approach for lateral lumbar interbody fusion Summary of Background Data : The introduction of laparoscopic techniques in 1993 has stimulated a great deal of discussion regarding the risks and benefits of such minimally invasive approaches. In many centers the anterior endoscopic approach to L5-S1 has become routine. However exposure at L4-5 can be much more difficult. MATERIALS AND METHODS: From 1997 to 1999 thirty eight patients were entered into a prospective study. These patients were all undergoing anterior interbody fusion at the L4-5 level. The patients were divided into two groups for analysis. Group I patients underwent anterior interbody fusion utilizing threaded interbody devices placed via laparoscopic lateral retroperitoneal approach. Group II patients underwent anterior lumbar interbody fusion using threaded interbody devices placed via a miniopen retroperitoneal approach. RESULTS: In Group I, Operation time was 48 minutes longer than Group II (p=0.035) but there were no significant statistical differences in bleeding amount and hospitalization period. Parethesia and tingling sensation of thigh were developed in two cases of Group I patients, one case of Group II patients but they were gradually diminished. In Group I, only one cage was inserted in five cases of patients (28%) who had an inadequate exposure of L4-5 area. However, all of the patients in Group II (100%) had an adequate exposure of L4-5 area. CONCLUSION: The surgical results of laparoscopic technique was not superior to miniopen technique.
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Humans , Hemorrhage , Hospitalization , Prospective Studies , Risk Assessment , Sensation , ThighABSTRACT
We investigated the mechanism of Cl- secretion by fluoroaluminate(AlF4-) and sodium orthovanadate(vanadate) using the human colonic T84 cell line. T84 cell monolayers grown on collagen-coated filters were mounted in Ussing chambers to measure short circuit current(ISC). Serosal addition of AlF4- or vanadate to T84 monolayers produced a sustained increase in ISC. Removal of Ca2+ from the serosal bathing solution partially inhibited AlF4-(-)and vanadate-induced ISC, and readministration of Ca2+ restored AlF4-(-)and vanadate-induced ISC. Carbachol application in the presence of forskolin, AlF4- or vanadate induced a synergistic increase of ISC. Forskolin and vanadate significantly increased cellular cAMP level, while carbachol and AlF4- did not. Carbachol, AlF4- and vanadate significantly increased [Ca2+]i. After Na+ in mucosal bathing solution was replaced with K+, and the mucosal membrane of T84 cell was permeabilized with amphotericin B, AlF4-, vanadate, and carbachol increased K+ conductance, but forskolin did not. After sodium chloride in serosal bathing solution was replaced with sodium gluconate and the serosal membrane was permeabilized with nystatin, forskolin, AlF4-, and vanadate increased Cl- conductance, but carbachol did not. AlF4-(-)induced ISC was remarkably inhibited by the pretreatment of pertussis toxin(2 micrograms/ml) for 2 hours. These results indicate that AlF4- and vanadate can increase Cl- secretion via simultaneous stimulation of Cl- channel and K+ channel in T84 cells. However, the AlF4- action is mostly attributed to stimulation of pertussis toxin-sensitive G-proteins, whereas the vanadate action mostly results from G protein-independent mechanisms.
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Humans , Aluminum/pharmacology , Amphotericin B/pharmacology , Carbachol/pharmacology , Cell Polarity , Cells, Cultured/drug effects , Chloride Channels/drug effects , Chlorides/physiology , Colon , Electrophysiology , Fluorine/pharmacology , Colforsin/pharmacology , GTP-Binding Proteins/physiology , Pertussis Toxin , Potassium/pharmacology , Potassium Channels/drug effects , Second Messenger Systems , Signal Transduction , Vanadates/pharmacology , Virulence Factors, Bordetella/pharmacologyABSTRACT
Since researches and clinical experiences up to now provide us with little helps in predicting what type of spinal tuberculosis will result in good healing by chemotherapy alone without kyphosis or with minimum kyphosis, it seems impractical to await natural healing with unsightly kyphosis. Therefore, my opinion is that surgery to achieve early cure and to correct and/or prevent kyphosis is desirable. The two-stage operation