Performance and healthcare analysis in elite sports teams using artificial intelligence: a scoping review.

Introduction: In competitive sports, teams are increasingly relying on advanced systems for improved performance and results. This study reviews the literature on the role of artificial intelligence (AI) in managing these complexities and encouraging a system thinking shift. It found various AI applications, including performance enhancement, healthcare, technical and tactical support, talent identification, game prediction, business growth, and AI testing innovations. The main goal of the study was to assess research supporting performance and healthcare. Methods: Systematic searches were conducted on databases such as Pubmed, Web of Sciences, and Scopus to find articles using AI to understand or improve sports team performance. Thirty-two studies were selected for review. Results: The analysis shows that, of the thirty-two articles reviewed, fifteen focused on performance and seventeen on healthcare. Football (Soccer) was the most researched sport, making up 67% of studies. The revised studies comprised 2,823 professional athletes, with a gender split of 65.36% male and 34.64% female. Identified AI and non-AI methods mainly included Tree-based techniques (36%), Ada/XGBoost (19%), Neural Networks (9%), K-Nearest Neighbours (9%), Classical Regression Techniques (9%), and Support Vector Machines (6%). Conclusions: This study highlights the increasing use of AI in managing sports-related healthcare and performance complexities. These findings aim to assist researchers, practitioners, and policymakers in developing practical applications and exploring future complex systems dynamics.

Analyzing ECG signals in professional football players using machine learning techniques.

Background: Football player's health is important, and preventing sudden cardiac arrest may be a critical issue. Professional football players have different ECG signals than the average population, yet there are considerable gaps in study whereas the general population has been extensively studied. Objectives: (a) Generate a reference and innovative resting 12-lead ECG database from 54 UEFA PRO level male football players from La Liga. This is a novel approach to cope the ECG and possible arrythmias in athletes. (b) Manage each XML athlete ECG data and develop a free-use program to visualize, denoise and filter the signal with the capacity to automate the labelling of the waves and save the reports. (c) Study the ECG wave shape and generate models through ML to analyse its utility to automate basic diagnosis. Methods: The dataset collection is based on a prospective observational cohort and includes 10 s, 12-lead ECGs and rhythm and condition labels for each athlete. Physiological sport arrhythmias, T-Wave shape and other findings were studied and labelled. ECG Visualizer was developed and used for 3 machine learning (ML) methods to automate sinus bradycardia arrhythmia diagnosis. Results: A dataset with 163 ECGs in XML format was collected comprising the Pro Football 12-lead Resting Electrocardiogram Database (PF12RED). "ECG Visualizer" software was developed, and ML was shown to be useful in detecting sinus bradycardia. Conclusions: The study demonstrates that AI and machine learning can detect simple arrhythmias with accuracy, also it provides a valuable dataset and a free software application.

Polymorphism in the pvcrt-o and pvmdr-1 genes of Plasmodium vivax associated with a better prognosis for malaria.

The early molecular identification of strains of Plasmodium vivax that have a worse prognosis is important to stratify the risk of complications and choice of conduct made by medical teams. Thus, the aim of the present study was to associate the presence of polymorphisms in the pvmdr-1 and pvcrt-o resistance genes of P. vivax in patients with better or worse prognosis. This cross-sectional epidemiological study was conducted based on data obtained from the records of 120 patients diagnosed with malaria in the Brazilian Amazon. The T958M and F1076L mutations of the pvmdr-1 gene had a frequency of 3.3 and 4.2%, respectively, and primo-infected patients had a 17 times greater chance of being infected with protozoa with the T958M mutation compared to patients with previous episodes. Regarding pvcrt-o, the C393T and T786C polymorphisms had a frequency of 14.2 and 3.3%, respectively, and self-declared white patients had a 3.1 times greater chance of being infected with protozoa with the C393T polymorphism. In addition, patients with this pvcrt-o polymorphism had lower concentrations of C-reactive protein, indicating a better prognosis. These data present clues of genetic indicators useful for assessing the virulence of the parasite and the prognosis of patients with vivax malaria.

Polymorphism in the pvcrt-o and pvmdr-1 genes of Plasmodium vivax associated with a better prognosis for malaria

@#The early molecular identification of strains of Plasmodium vivax that have a worse prognosis is important to stratify the risk of complications and choice of conduct made by medical teams. Thus, the aim of the present study was to associate the presence of polymorphisms in the pvmdr-1 and pvcrt-o resistance genes of P. vivax in patients with better or worse prognosis. This cross-sectional epidemiological study was conducted based on data obtained from the records of 120 patients diagnosed with malaria in the Brazilian Amazon. The T958M and F1076L mutations of the pvmdr-1 gene had a frequency of 3.3 and 4.2%, respectively, and primo-infected patients had a 17 times greater chance of being infected with protozoa with the T958M mutation compared to patients with previous episodes. Regarding pvcrt-o, the C393T and T786C polymorphisms had a frequency of 14.2 and 3.3%, respectively, and self-declared white patients had a 3.1 times greater chance of being infected with protozoa with the C393T polymorphism. In addition, patients with this pvcrt-o polymorphism had lower concentrations of C-reactive protein, indicating a better prognosis. These data present clues of genetic indicators useful for assessing the virulence of the parasite and the prognosis of patients with vivax malaria.

Dynamic expression of Ralstonia solanacearum virulence factors and metabolism-controlling genes during plant infection.

BACKGROUND: Ralstonia solanacearum is the causal agent of bacterial wilt, a devastating plant disease responsible for serious economic losses especially on potato, tomato, and other solanaceous plant species in temperate countries. In R. solanacearum, gene expression analysis has been key to unravel many virulence determinants as well as their regulatory networks. However, most of these assays have been performed using either bacteria grown in minimal medium or in planta, after symptom onset, which occurs at late stages of colonization. Thus, little is known about the genetic program that coordinates virulence gene expression and metabolic adaptation along the different stages of plant infection by R. solanacearum. RESULTS: We performed an RNA-sequencing analysis of the transcriptome of bacteria recovered from potato apoplast and from the xylem of asymptomatic or wilted potato plants, which correspond to three different conditions (Apoplast, Early and Late xylem). Our results show dynamic expression of metabolism-controlling genes and virulence factors during parasitic growth inside the plant. Flagellar motility genes were especially up-regulated in the apoplast and twitching motility genes showed a more sustained expression in planta regardless of the condition. Xylem-induced genes included virulence genes, such as the type III secretion system (T3SS) and most of its related effectors and nitrogen utilisation genes. The upstream regulators of the T3SS were exclusively up-regulated in the apoplast, preceding the induction of their downstream targets. Finally, a large subset of genes involved in central metabolism was exclusively down-regulated in the xylem at late infection stages. CONCLUSIONS: This is the first report describing R. solanacearum dynamic transcriptional changes within the plant during infection. Our data define four main genetic programmes that define gene pathogen physiology during plant colonisation. The described expression of virulence genes, which might reflect bacterial states in different infection stages, provides key information on the R. solanacearum potato infection process.

Technology at the service of surgery in a new technique of autotransplantation by guided surgery: a case report.

BACKGROUND: The aim of this case report was to use a surgical technique for autotransplantation of tooth using virtually planned 3D printed surgical templates for guided osteotomy preparation of the recipient of donor tooth. CASE PRESENTATION: An 18-year-old male patient received autotransplantation of the right mandibular third molar to replace an included right second molar. This procedure was based on guided implant surgery methods by superimposition of DICOM files and 3D data sets of the jaws. In order to design a 3D-printed template with the aid of a fully digital workflow; the third molar was conserved in PRGF during the surgical procedure and the tooth socket was prepared with a template and the help of a 3D-printed donor tooth copy in order to prevent iatrogenic damage to the donor tooth. This template and replica were manufactured using 3D-printing techniques. The transplanted tooth was placed in infra-occlusion and fixed with a suture splint and root canal therapy was performed 15 days later. The intervention was be accomplished by performing preplanned virtual transplantations with guided osteotomies to ensure accurate donor tooth placement in the new recipient site. The 24 months follow-up showed physiological clinical and radiologic results compatible with healing periradicular tissues. CONCLUSIONS: This approach enables the planning and production of a 3D printed surgical template using the latest diagnostic methods and techniques of guided implant surgery. These accurate virtually predesigned surgical templates and printed analogues of the donor tooth could facilitate autotransplantation, ensuring an atraumatic surgical protocol.

Influence of intestinal resections on biliary composition and liver ultrastructure.

BACKGROUND & AIMS: Among the multiple factors that can influence bile acid (BA) metabolism and biliary poles, intestinal resections could do so directly (through absorption alterations) and/or indirectly (bacterial translocation, alterations of the hepato-intestinal hormonal axis). Our aim was to study the influence of partial intestinal resections on bile metabolism and biliary structure in an animal model. METHODS: Dunkin Hartley guinea pigs were used: Group A with only jejunum resection, Group B with only ileum resection, and Group CtrG, the control group. They were monitored for 21 days, with oral nutrition. At the end of this period, samples of the bile were taken from the hepatic duct and vena cava. After liver perfusion, samples of hepatic tissue were taken for optical and electron microscopy. RESULTS: There was a significant decrease in serum cholesterol in both GA & GB. The levels of biliary phospholipids (Ph) and cholic acid decreased only in Group B, but not ketolithocholic acid nor total conjugated BA. Slight liver macrosteatosis was observed in GB, but there were no changes in the biliary poles or canaliculi in the electron microscopy samples. CONCLUSIONS: Ileum resections affect the biliary Ph and BA composition, which could increase bile lithogenicity but do not seem to affect biliary duct anatomy.

Cambios en la composición de la grasa y en la histomorfología del hígado tras resecciones intestinales parciales / Changes in the fat composition and histomorphology of the liver after partial intestinal resections

Introducción y objetivos: Teóricamente, las resecciones intestinales parciales podrían afectar al hígado a través de la absorción de nutrientes y de la circulación enterohepática de los ácidos biliares, por las posibles alteraciones sobre el complejo hormonal duodeno-bilo-hepato-pancreático y por los efectos secundarios de la translocación bacteriana intestinal. Estos mecanismos fisiopatológicos pueden sumarse modificando la composición y la histomorfología hepática, específicamente la composición de la grasa hepática. El objetivo de este trabajo experimental es conocer mas profunda y concretamente las alteraciones secundarias a las resecciones intestinales parciales. M & M: Utilizamos 26 cobayos divididos en 3 grupos: Grupo A (n=10), con resección yeyunal; Grupo B (n=10) con resección ileal y Grupo control (GCtr, n=6). Los animales fueron nutridos oralmente durante 21 días. Al final se obtuvieron muestras para analizar la bioquímica sérica y se homogeneizó el tejido hepático para estudio de la grasa hepática (grasa total, proporciones de lípidos neutros y polares y proporciones de fosfolípidos y de ácidos grasos). Se tomaron muestras para estudio histomorfológico. Resultados: Los animales con resección yeyunal e ileal presentaron disminución del colesterol sérico, del peso de la gota grasa/g tejido y del cociente w6/w3 y del C18-2/C18-3, sin alteraciones en la distribución de fosfolípidos. Se halló macroesteatosis y disminución de la densidad mitocondrial en los animales del G-B. Conclusiones: las resecciones intestinales parciales, especialmente las ileales sí parecen influir a corto plazo en la composición de la grasa estructural del hígado. Estos cambios en los lípidos estructurales y en la histomorfología pudieran ser compensados a medio plazo. Estos hallazgos pueden ayudarnos a entender mejor la relación hepato-intestinal (AU)

Background & Aims: The partial resection of the intestine could affect the liver through alteration in nutrient absorption, loss of the bile acids via the enterohepatic circulation, alterations in the hepato-duodenobilio-pancreatic hormonal complex as well as a bacterial translocation. All these factors could sum up and induce changes in the tissue composition and in hepatic histomorphology. The aim of this experimental study is to acquire a profound knowledge of these hepatic alterations after the partial intestinal resection. M & M: 26 Dunkin Hartley guinea pigs were used: Group A with only jejunum resection, Group B with only ileum resection, and Group CtrG, the control group. They were monitored for 21 days. At the end, blood samples were taken for serum biochemical analysis. Samples of hepatic tissue were taken for histomorphological optical study. All the rest liver was homogenize and the fat liver composition was analysed (total fat, lipid fractions, phospholipids fractions and fatty acids of the liver). Results: There was a significant decrease in serum cholesterol in both GA & GB. There was also a decrease of the total fat/g liver tissue, without changes in the phospholipids fraction and decreases in the C18-2/C18-3 and w6/w3 ratios. There was a minimal macrosteatosis and reduction of the number of mitochondries in GB. Conclusions: the partial intestinal resection, specially ileum resections, looks like influence the structural fat liver composition. However, these changes could be compensated for by the liver and oral nutrition. These findings help us to improve the liver-intestinal relationships (AU)

[Changes in the fat composition and histomorphology of the liver after partial intestinal resections]. / Cambios en la composición de la grasa y en la histomorfología del hígado tras resecciones intestinales parciales.

BACKGROUND & AIMS: The partial resection of the intestine could affect the liver through alteration in nutrient absorption, loss of the bile acids via the enterohepatic circulation, alterations in the hepato-duodenobilio-pancreatic hormonal complex as well as a bacterial translocation. All these factors could sum up and induce changes in the tissue composition and in hepatic histomorphology. The aim of this experimental study is to acquire a profound knowledge of these hepatic alterations after the partial intestinal resection. M & M: 26 Dunkin Hartley guinea pigs were used: Group A with only jejunum resection, Group B with only ileum resection, and Group CtrG, the control group. They were monitored for 21 days. At the end, blood samples were taken for serum biochemical analysis. Samples of hepatic tissue were taken for histomorphological optical study. All the rest liver was homogenize and the fat liver composition was analysed (total fat, lipid fractions, phospholipids fractions and fatty acids of the liver). RESULTS: There was a significant decrease in serum cholesterol in both GA & GB. There was also a decrease of the total fat/g liver tissue, without changes in the phospholipids fraction and decreases in the C18-2/C18-3 and w6/w3 ratios. There was a minimal macrosteatosis and reduction of the number of mitochondries in GB. CONCLUSIONS: the partial intestinal resection, specially ileum resections, looks like influence the structural fat liver composition. However, these changes could be compensated for by the liver and oral nutrition. These findings help us to improve the liver-intestinal relationships.

Anti-inflammatory activity of flavonoids from Eupatorium arnottianum.

Three anti-inflammatory compounds: nepetin, jaceosidin and hispidulin have been isolated and identified from Eupatorium arnottianum Griseb. dichloromethane extract. Nepetin reduced the TPA mouse ear edema by 46.9% and jaceosidin by 23.2% (1mg/ear). Both compounds inhibited the NF kappaB induction by 91 and 77%, respectively. Furthermore phytochemical analysis of the ethanol extract has led to the identification of eriodictyol, hyperoside, rutin, caffeic and chlorogenic acids. All these compounds are reported for the first time in this species. The finding of topical antiinflammatory activity exerted by Eupatorium arnottianum extract and the identification of active principles could support the use of this plant for the treatment of inflammatory affections.

Bioguided extraction of polyphenols from grape marc by using an alternative supercritical-fluid extraction method based on a liquid solvent trap.

The biological activity of polyphenols extracted from grape marc was studied with a view to finding a new use for this winery waste. Polyphenols were extracted by using an alternative supercritical-fluid extraction method based on the use of a liquid trap that allows extracted polyphenols to be retained in a saline buffer, thus avoiding the need for the organic solvent required to elute polyphenols from a solid trap. The major extraction variables influencing the performance of the liquid trap (viz. CO(2) modifier content, flow-rate, extraction time and trap volume) were optimized. The proposed method was applied to the supercritical-fluid extraction extraction of 0.3 g grape marc with CO(2) modified with 3% methanol at 350 bar at 50 degrees C (CO(2) density 0.9 g mL(-1)) for 20 min, using a liquid flow-rate of 0.9 mL min(-1). The polyphenol extracts thus obtained exhibited cytotoxic effects that induced apoptosis in tumour cells.

Soluble dipeptidyl peptidase IV (CD-26) in serum of patients with colorectal carcinoma.

Dipeptidyl peptidase IV(DPPIV) or CD26 is a widely expressed ectopeptidase with functions in immune response such as the activation of T lymphocytes. It is expressed in the colon mucosal epithelium only when this becomes malignant. The aim of our study is to ascertain the soluble DPPIV/CD26 levels in the serum of patients with colorectal carcinoma, compared with the levels in healthy individuals. From an accrual of 70 healthy individuals and 99 patients diagnosed with colorectal adenocarcinoma, the levels of soluble DPPIV/CD26 were defined by colored enzymatic spectrophotometric response on the Gly-Pro-Paranitroaniline substrate. The patients diagnosed with colorectal cancer have a higher CD26 level than healthy subjects (p<0.05). Of these patients, those with metastatic colorectal disease have a significantly higher soluble CD26 level (p<0.01). It has also been found that patients with high LDH (lactatodeshidrogenase) and CEA (carcinoembrionary antigen) levels show higher CD26 levels (p<0.01) than patients with normal levels of such carcinoma markers.

Cellular redox state and activating protein-1 are involved in ascorbate effect on calcitriol-induced differentiation.

Ascorbate has been related to the differentiation of several mesenchymal cells including haematopoietic cells. We have previously demonstrated that ascorbate enhances the activity of 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2D3) on monocytic differentiation of HL-60 cells. Here, we show that ascorbate-mediated modification of cellular redox state and AP-1 (activating protein-1) DNA binding during early phases are related to the enhancing effect of ascorbate on differentiation. Ascorbate, but not its fully oxidized form, dehydroascorbate, or an ascorbate analogue with a low rate of oxidation, ascorbate-2-phosphate, enhanced the differentiation induced by 1 alpha,25(OH)2D3, modified cytosolic reactive oxygen species levels and mitochondrial redox potential (delta psi m), and modulated AP-1 DNA binding in HL-60 cells. Ascorbate itself increased AP-1 binding to DNA in noninduced cells, whereas it inhibited AP-1 binding in 1 alpha,25(OH)2D3-induced cells. However, ascorbate increased the mRNA levels of c-jun, junB, and c-fos in 1 alpha,25(OH)2D3-induced cells. Taken together, these results suggest that the enhancing effect of ascorbate on HL-60 differentiation induced by 1 alpha, 25(OH)2D3 is related to its effect on the cellular redox state and the modulation of AP-1 activity.

Ingenol esters induce apoptosis in Jurkat cells through an AP-1 and NF-kappaB independent pathway.

BACKGROUND: Ingenol derivatives have received constant and multidisciplinary attention on account of their pleiotropic pattern of biological activity. This includes activation of protein kinase C (PKC), tumour-promotion, anticancer, and anti-HIV properties, and the possibility of dissecting co-cancerogenic and clinically useful activities has been demonstrated. Certain ingenol esters show powerful anticancer activity, and a structure-activity relationship model to discriminate between their apoptotic and non-apoptotic properties has been developed. RESULTS: The polyhydroxylated southern region of ingenol was selectively modified, using the anticancer and PKC activator ingenol 3,20-dibenzoate (IDB) as a lead compound. The evaluation of IDB analogues in apoptosis assays showed strict structure-activity relationships, benzoylation of the 20-hydroxyl being required to trigger apoptosis through a pathway involving caspase-3 and occurring at the specific cell cycle checkpoint that controls the S-M phase transition. Conversely, a study on the activation of the PKC-dependent transcription factors AP-1 and NF-kappaB by IDB analogues showed significant molecular flexibility, including tolerance to changes at the 3- and 20-hydroxyls. IDB-induced apoptosis was independent of activation of PKC, since it was not affected by treatment with the non-isoform-selective PKC inhibitor GF 109230X0. CONCLUSIONS: Remarkable deviations from the tumour-promotion pharmacophore were observed for both the apoptotic and the PKC-activating properties of IDB analogues, showing that ingenol is a viable template to selectively target crucial pathways involved in tumour promotion and development. Since the apoptotic and the PKC-activating properties of ingenoids are mediated by different pathways and governed by distinct structure-activity relationships, it is possible to dissect them by suitable chemical modification. In this context, the esterification pattern of the 5- and 20-hydroxyls is critical.

New glycosides from Capsicum annuum L. var. acuminatum. Isolation, structure determination, and biological activity.

Investigation of polar extracts from ripe fruits of Capsicum annuum L. var. acuminatum yielded three new glycosides, capsosides A (1) and B (2) and capsianoside VII (3), along with seven known compounds (4-10). The chemical structures were elucidated mainly by extensive nuclear magnetic resonance methods and mass spectrometry, and the biological activities of icariside E(5) (4) were tested by different assays. Icariside E(5), in contrast to capsaicin, neither induces an increase in the intracellular levels of reactive oxygen species nor affects the mitochondria permeability transition, and it does not signal through the vanilloid receptor type 1. Interestingly, this compound protects Jurkat cells from apoptosis induced by the oxidative stress mediated by serum withdrawal. These results suggest that icariside E(5) may have antioxidant properties that strengthen the importance of peppers in the Mediterranean diet.

Evisceración vaginal. aportación de dos casos y revisión de la bibliografía / Vaginal evisceration. Report of two cases and review of the literature

La evisceración vaginal es una patología extremadamente rara que suele aparecer en mujeres posmenopáusicas, asociada a cirugía ginecológica e hipoestrogenismo. Excepcionalmente puede ocurrir en mujeres jóvenes tras violación o introducción de cuerpos extraños. En este trabajo hacemos una revisión bibliográfica y presentamos 2 casos ocurridos recientemente en nuestro servicio; analizamos los factores desencadenantes y exponemos la manera de prevenirlos, así como su tratamiento (AU)

Phorboid 20-homovanillates induce apoptosis through a VR1-independent mechanism.

BACKGROUND: Vanilloids, such as capsaicin and resiniferatoxin (RTX), are recognized at the cell surface by vanilloid receptor type 1 (VR1), which has recently been cloned. VR1 mediates the effects of capsaicin and RTX in VR1-expressing cells, but vanilloids can induce apoptosis through a pathway not mediated by VR1. Phorboid 20-homovanillates can be used to investigate cell death induced by vanilloids. RESULTS: 12,13-Diacylphorbol-20 homovanillates were prepared by the sequential esterification of the natural polyol. Phorbol 12-phenylacetate 13-acetate 20-homovanillate (PPAHV) induced apoptosis in Jurkat cells to the same extent as RTX. Apoptosis was preceded by an increase in intracellular reactive oxygen species and by the loss of mitochondrial transmembrane potential. PPAHV-induced apoptosis was mediated by a pathway involving caspase-3 activation and was initiated at the S phase of the cell cycle. The cell-death pathway triggered by VR1 activation was studied in 293T cells transfected with the cloned rat vanilloid receptor. In this system, capsaicin and PPAHV induced cell death by an apparent necrotic mechanism, which was selectively inhibited by the competitive vanilloid receptor antagonist capsazepine. Interestingly, phorbol-12, 13-bisnonanoate-20-homovanillate, an analogue of PPAHV, induced cell death in VR1-transfected cells but could not trigger apoptosis in the Jurkat cell line. CONCLUSIONS: Vanilloids can induce cell death through different signalling pathways. The cell death induced in a VR1-independent manner has the hallmark of apoptosis, whereas the cell death mediated by vanilloids binding to VR1 is seemingly necrotic. Phorboid homovanillates that have antitumour and anti-inflammatory activities but lack the undesirable side effects of the natural vanilloids could be developed as potential drugs.

Hydroxyurea inhibits the transactivation of the HIV-long-terminal repeat (LTR) promoter.

HIV-1 gene expression is regulated by the promoter/enhancer located within the U3 region of the proviral 5' LTR that contains multiple potential cis-acting regulatory sites. Here we describe that the inhibitor of the cellular ribonucleoside reductase, hydroxyurea (HU), inhibited phorbol myristate acetate- or tumour necrosis factor-alpha-induced HIV-1-LTR transactivation in both lymphoid and non-lymphoid cells in a dose-dependent manner within the first 6 h of treatment, with a 50% inhibitory concentration of 0.5 mM. This inhibition was found to be specific for the HIV-1-LTR since transactivation of either an AP-1-dependent promoter or the CD69 gene promoter was not affected by the presence of HU. Moreover, gel-shift assays in 5.1 cells showed that HU prevented the binding of the NF-kappaB to the kappaB sites located in the HIV-1-LTR region, but it did not affect the binding of both the AP-1 and the Sp-1 transcription factors. By Western blots and cell cycle analyses we detected that HU induced a rapid dephosphorylation of the pRB, the product of the retinoblastoma tumour suppressor gene, and the cell cycle arrest was evident after 24 h of treatment. Thus, HU inhibits HIV-1 promoter activity by a novel pathway that implies an inhibition of the NF-kappaB binding to the LTR promoter. The present study suggests that HU may be useful as a potential therapeutic approach for inhibition of HIV-1 replication through different pathways.

Involvement of mitochondria and caspase-3 in ET-18-OCH(3)-induced apoptosis of human leukemic cells.

The induction of cell death in leukemic HL-60 cells by the ether lipid 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-18-OCH(3); edelfosine) followed the typical apoptotic changes in ultrastructural morphology, including blebbing, chromatin condensation, nuclear membrane breakdown and extensive vacuolation. Using a cytofluorimetric approach, we found that ET-18-OCH(3) induced disruption of the mitochondrial transmembrane potential (DeltaPsi(m)) followed by production of reactive oxygen species (ROS) and DNA fragmentation in leukemic cells. ET-18-OCH(3) also induced caspase-3 activation in human leukemic cells, as assessed by cleavage of caspase-3 into the p17 active form and cleavage of the caspase-3 substrate poly(ADP-ribose) polymerase (PARP). ET-18-OCH(3) analogues unable to induce apoptosis failed to disrupt DeltaPsi(m) and to activate caspase-3. ET-18-OCH(3)-resistant Jurkat cells generated from sensitive Jurkat cells showed no caspase-3 activation and did not undergo DeltaPsi(m) disruption upon ET-18-OCH(3) incubation. Cyclosporin A partially inhibited DeltaPsi(m) dissipation, caspase activation and apoptosis in ET-18-OCH(3)-treated leukemic cells. Overexpression of bcl-2 by gene transfer prevented DeltaPsi(m) collapse, ROS generation, caspase activation and apoptosis in ET-18-OCH(3)-treated leukemic T cells. Pretreatment with the caspase inhibitor Z-Asp-2, 6-dichlorobenzoyloxymethylketone prevented ET-18-OCH(3)-induced PARP proteolysis and DNA fragmentation, but not DeltaPsi(m) dissipation. ET-18-OCH(3) did not affect the expression of caspases and bcl-2-related genes. ET-18-OCH(3)-induced apoptosis did not require protein synthesis. Our data indicate that DeltaPsi(m) dissipation and caspase-3 activation are critical events of the apoptotic cascade triggered by the antitumor ether lipid ET-18-OCH(3), and that the sequence of events in the apoptotic action of ET-18-OCH(3) on human leukemic cells is: DeltaPsi(m) disruption, caspase-3 activation and internucleosomal DNA degradation.