Case report: Neuronal intranuclear inclusion disease presenting with acute encephalopathy.

Neuronal intranuclear inclusion disease (NIID), a neurodegenerative disease previously thought to be rare, is increasingly recognized despite heterogeneous clinical presentations. NIID is pathologically characterized by ubiquitin and p-62 positive intranuclear eosinophilic inclusions that affect multiple organ systems, including the brain, skin, and other tissues. Although the diagnosis of NIID is challenging due to phenotypic heterogeneity, a greater understanding of the clinical and imaging presentations can improve accurate and early diagnosis. Here, we present three cases of pathologically proven adult-onset NIID, all presenting with episodes of acute encephalopathy with protracted workups and lengthy time between symptom onset and diagnosis. Case 1 highlights challenges in the diagnosis of NIID when MRI does not reveal classic abnormalities and provides a striking example of hyperperfusion in the setting of acute encephalopathy, as well as unique pathology with neuronal central chromatolysis, which has not been previously described. Case 2 highlights the progression of MRI findings associated with multiple NIID-related encephalopathic episodes over an extended time period, as well as the utility of skin biopsy for antemortem diagnosis.

Morphological adaptations in response to chronic exercise across musculoskeletal tissues: a systematic review / Adaptaciones morfológicas en respuesta al ejercicio crónico en los tejidos osteomusculares: una revisión sistemática

León, F., Mestre. A., Priego, L., & Vera, J.C. (2023). Morphological adaptations in response to chronic exercise across musculoskeletal tissues: a systematic review. PENSAR EN MOVIMIENTO: Revista de Ciencias del Ejercicio y la Salud, 21(1), 1-28. To date, there is no systematic review that summarizes the morphological adaptations of the musculoskeletal system in response to chronic exercise. This systematic review selected original articles published in English between 2000 and 2020, with a clear exercise intervention and presenting a morphological change in the tissue under study, and covering human participants irrespective of age, gender or health condition. In total, 2819 records were identified. After removal of duplicates, title and abstract screening and full-text review, 67 records were included in the final analysis (6 for inter-vertebral disc, 6 for cartilage, 36 for bone, 2 for ligament, 9 for tendon and 7 for muscle). The most used interventions were aerobic, resistance, and plyometric exercise. Population ranged from children and healthy active people to individuals with a health condition. In conclusion, as a response to chronic exercise there are morphological adaptations in the tissues of the musculoskeletal system which vary from increased stiffness to an increase in cross-sectional area. Although tissues can adapt, several questions still linger, such as optimal dose and type of exercise, whether adaptations can occur in an injured tissue, and functional implications of these adaptations. Future research should address these questions.

León, F., Mestre. A., Priego, L. y Vera, J.C. (2023). Adaptaciones morfológicas en respuesta al ejercicio crónico en los tejidos osteomusculares: una revisión sistemática. PENSAR EN MOVIMIENTO: Revista de Ciencias del Ejercicio y la Salud, 21(1), 1-28. Hasta la fecha, no existe una revisión sistemática que resuma las adaptaciones morfológicas del sistema osteomuscular en respuesta al ejercicio crónico. Esta revisión sistemática seleccionó artículos originales, con fecha de publicación de 2000 a 2020, idioma de publicación en inglés, con una clara intervención de ejercicio y que presentaron un cambio morfológico en el tejido estudiado. Participantes humanos independientemente de la edad, el género o condición de salud. Se identificaron 2819 registros. Después de eliminar los duplicados, la selección de títulos y resúmenes y la revisión de texto completo, se incluyeron 67 registros en el análisis final (6 para disco intervertebral, 6 para cartílago, 36 para hueso, 2 para ligamento, 9 para tendón y 7 para músculo). Los resultados destacan que las intervenciones más utilizadas fueron ejercicio aeróbico, contra resistencia y pliométrico. La población abarcó desde niños y personas sanas activas hasta personas con alguna condición de salud. Se concluye que como respuesta al ejercicio crónico existen adaptaciones morfológicas en los tejidos del sistema musculoesquelético, que pueden variar desde un aumento de rigidez hasta un aumento de área. Aunque los tejidos pueden adaptarse, aún quedan varias preguntas, como la dosis y tipo de ejercicio óptimo, si pueden ocurrir adaptaciones en un tejido lesionado y las implicaciones funcionales de estas adaptaciones. La investigación futura debe abordar estas preguntas.

León, F., Mestre. A., Priego, L. e Vera, J.C. (2023). Adaptações morfológicas em resposta ao exercício crônico nos tecidos osteomusculares: uma revisão sistemática. PENSAR EN MOVIMIENTO: Revista de Ciencias del Ejercicio y la Salud, 21(1), 1-28. Até o momento, não há uma revisão sistemática que resuma as adaptações morfológicas do sistema osteomuscular em resposta ao exercício crônico. Esta revisão sistemática selecionou artigos originais, com data de publicação de 2000 a 2020, idioma de publicação em inglês, com clara intervenção de exercícios e que apresentaram alteração morfológica no tecido estudado. Participantes humanos, independentemente da idade, sexo ou condição de saúde. Foram identificados 2.819 registros. Após eliminar os artigos duplicados, triagem de título e resumo e revisão do texto completo, 67 registros foram incluídos na análise final (6 para disco intervertebral, 6 para cartilagem, 36 para osso, 2 para ligamento, 9 para tendão e 7 para músculo). Os resultados destacam que as intervenções mais utilizadas foram exercícios aeróbicos, resistidos e pliométricos. A população variou de crianças e pessoas saudáveis ativas a pessoas com alguma condição de saúde. Conclui-se que, em resposta ao exercício crônico, ocorrem adaptações morfológicas nos tecidos do sistema musculoesquelético, que podem variar desde um aumento de rigidez até um aumento de área. Embora os tecidos possam se adaptar, várias questões permanecem, como a dose ideal e o tipo de exercício, se adaptações podem ocorrer no tecido lesado e as implicações funcionais dessas adaptações. Pesquisas futuras devem abordar essas questões.

Pain Relief After Allogenic Stem Cell Disc Therapy.

BACKGROUND: Treatment of intermediate-stage painful degenerative disc disease is controversial, with few reliable options. Allogenic mesenchymal stem cells (MSCs)are an alternative to autologous stem cell transplantation. Allogeneic MSCs in the treatment of discogenic low back pain have some practical advantages, ranging from availability to ease of treatment in a procedure-room setting. OBJECTIVES: To assess the efficacy and safety of allogenic MSC injection into painful lumbar intervertebral discs and associated clinical outcomes. STUDY DESIGN: Retrospective observational cohort study. SETTING: Private practice. METHODS: There were 33 patients: 15 women and 18 men with an average age of 47.6 years. The patients' average follow-up was 26.88 months Patients were treated with intradiscal injection of approximately 5 million allogeneic polyclonal MSCs in 1% hyaluronic acid derived from immunoselected umbilical cord stem cells. Patients were monitored for adverse event reactions. Clinical outcomes were assessed with reductions in the reported Visual Analog Scale (VAS) for back pain, the Oswestry Disability Index (ODI) scores, and the use of the modified Macnab criteria. RESULTS: No patient required any additional treatments for low back pain stemming from the level treated with MSC injections. At a 2-year follow-up, the average VAS low back score reduction was 6.565 ± 1.619 and 38.333 ± 14.865 for the ODI (P < 0.001). Reported Macnab outcomes were excellent in 11 patients (33.3%), good in 19 (57.6%), and fair in 3 (9.1%). LIMITATIONS: Our observational study is limited by patient selection, hindsight bias, and low patient numbers. CONCLUSION: The results of our feasibility study suggest that the injection of allogeneic MSCs to treat patients with painful intermediate-stage degenerative disc disease has merit. No adverse reactions were observed. The authors recommend further study in a randomized prospective study setting with a placebo control group or a natural history study group of patients to solidify this research.

Caracterización clínica y metabólica de pacientes con diagnóstico de urolitiasis atendidos en una clínica de cuarto nivel de Barranquilla, Colombia / Clinical and metabolic characterization of patients with a diagnosis of urolithiasis in a four-level clinic in the city of Barranquilla

Resumen Introducción: la urolitiasis es una enfermedad frecuente de la cual en Colombia se han publicado estudios previos; sin embargo, estos no comparan las características sociodemográficas y clínicas de los pacientes con las comorbilidades y los factores predisponentes de litiasis como hiperuricemia, hipertensión arterial (HTA), obesidad y enfermedad renal crónica (ERC). Objetivos: caracterizar clínica y metabólicamente los pacientes con diagnóstico de urolitiasis atendidos en una clínica de cuarto nivel de Barranquilla, Colombia, en el año 2019. Materiales y métodos: se realizó un estudio observacional, descriptivo y transversal en 49 pacientes con base en el estudio de fichas clínicas Resultados: el 53,1 % de los participantes eran hombres y las medianas de edad y de índice de masa corporal (IMC) fueron 58 años y 26,4 kg/m2, respectivamente. Algunas de las comorbilidades identificadas fueron, en orden de frecuencia, HTA (69,4 %), ERC (36,7 %), infección de vías urinarias recurrente (24,5 %), hiperuricemia (44,9 %), hipercalcemia (16,3 %) e hiperfosfatemia (12,2 %). Los tipos de cristal encontrados fueron oxalato (20,4 %), urato (12,2 %), mezcla de oxalato y urato (4,1 %), fosfato (4,1 %), hipercalciuria e hiperoxaluria (38,8 %), hiperuricosuria e hipocalciuria (18,4 %) y hipofosfaturia o hipofosfaturia (4,1 %). Asimismo, la hiperuricemia se asoció a edad (p=0,028), ERC (p=0,026), medicamentos antihipertensivos (p=0,022), posición del cálculo en cáliz renal (p=0,012), hiperparatiroidismo (p=0,007), depuración de creatinina (p=0,046) e hipercalciuria (p=0,049). El IMC ≥30 se asoció con ERC estadio 5 (p=0,025), diálisis (p=0,025) e hiperoxaluria (p=0,021). Conclusiones: en la población analizada se evidenció una frecuencia significativa de ERC, hiperuricemia, obesidad e HTA.

Abstract Introduction: Urolithiasis is a disease with high frequency and our environment is no exception. Previous studies have been published in Colombia, however, these do not compare the sociodemographic and clinical characteristics of patients with comorbidities and predisposing factors for lithiasis such as they are hyperuricemia, high blood pressure, obesity, and chronic kidney disease (CKD). Objectives: To characterize clinically and metabolically the patients diagnosed with urolithiasis in a fourth-level clinic in the city of Barranquilla in 2019. Methods: Observational, descriptive, cross-sectional study. In 49 patients, based on the study of clinical records Results: The median age was 58 years, the male sex in 53.1 %. The median body mass index was 26.4 Kg / m2. High blood pressure was identified in 69.4 %, chronic kidney disease (CKD) in 36.7 %, recurrent urinary tract infection in 24.5 %. Hyperuricemia in 44.9 %, hypercalcemia in 16.3 % and hyperphosphatemia in 12.2 %. The crystal types were oxalate in 20.4 %, urate in 12.2 %, mixture of the previous ones in 4.1 % and in the same proportion phosphate. Hypercalciuria and hyperoxaluria in 38.8 %, hyperuricosuria and hypocalciuria in 18.4 %, while hyperphosphaturia or hypophosphaturia in 4.1 %. Hyperuricemia was associated with age (p = 0.028), CKD (p = 0.026), antihypertensive drugs (p = 0.022), the position of the stone in the renal calyx (p = 0.012), hyperparathyroidism (p = 0.007), creatinine clearance (p = 0.046) and hypercalciuria (p = 0.049). BMI ≥30 was associated with stage 5 CKD (p = 0.025), dialysis (p = 0.025), and hyperoxaluria (p = 0.021). Conclusions: A significant frequency of CKD, hyperuricemia, obesity and hypertension was evidenced in patients with urolithiasis.

The Role of Chitosan as a Possible Agent for Enteric Methane Mitigation in Ruminants.

Livestock production is a main source of anthropogenic greenhouse gases (GHG). The main gases are CH4 with a global warming potential (GWP) 25 times and nitrous oxide (N2O) with a GWP 298 times, that of carbon dioxide (CO2) arising from enteric fermentation or from manure management, respectively. In fact, CH4 is the second most important GHG emitted globally. This current scenario has increased the concerns about global warming and encouraged the development of intensive research on different natural compounds to be used as feed additives in ruminant rations and modify the rumen ecosystem, fermentation pattern, and mitigate enteric CH4. The compounds most studied are the secondary metabolites of plants, which include a vast array of chemical substances like polyphenols and saponins that are present in plant tissues of different species, but the results are not consistent, and the extraction cost has constrained their utilization in practical animal feeding. Other new compounds of interest include polysaccharide biopolymers such as chitosan, mainly obtained as a marine co-product. As with other compounds, the effect of chitosan on the rumen microbial population depends on the source, purity, dose, process of extraction, and storage. In addition, it is important to identify compounds without adverse effects on rumen fermentation. The present review is aimed at providing information about chitosan for dietary manipulation to be considered for future studies to mitigate enteric methane and reduce the environmental impact of GHGs arising from livestock production systems. Chitosan is a promising agent with methane mitigating effects, but further research is required with in vivo models to establish effective daily doses without any detrimental effect to the animal and consider its addition in practical rations as well as the economic cost of methane mitigation.

Data on SVCT2 transporter expression and localization in cancer cell lines and tissues.

The data presented in this article are related to the research paper entitled "Increased expression of mitochondrial sodium-coupled ascorbic acid transporter-2 (mitSVCT2) as a central feature in breast cancer", available in Free Radical Biology and Medicine Journal [1]. In this article, we examined the SVCT2 transporter expression in various breast cancer cell lines using RT-PCR and Western blot assays. In addition, we analyzed the subcellular localization of SVCT2 by immunofluorescence colocalization assays and cellular fractionation experiments. Finally, an analysis of different cancer tissue microarrays immunostained for SVCT2 and imaged by The Human Protein Atlas (https://www.proteinatlas.org) is presented.

GLUT1 and GLUT8 support lactose synthesis in Golgi of murine mammary epithelial cells.

The mammary gland increases energy requirements during pregnancy and lactation to support epithelial proliferation and milk nutrients synthesis. Lactose, the principal carbohydrate of the milk, is synthetized in the Golgi of mammary epithelial cells by lactose synthase from glucose and UPD galactose. We studied the temporal changes in the expression of GLUT1 and GLUT8 in mammary gland and their association with lactose synthesis and proliferation in BALB/c mice. Six groups were used: virgin, pregnant at 2 and 17 days, lactating at 2 and 10 days, and weaning at 2 days. Temporal expression of GLUT1 and GLUT8 transporters by qPCR, western blot and immunohistochemistry, and its association with lactalbumin, Ki67, and cytokeratin 18 within mammary tissue was studied, along with subcellular localization. GLUT1 and GLUT8 transporters increased their expression during mammary gland progression, reaching 20-fold increasing in GLUT1 mRNA at lactation (p < 0.05) and 2-fold at protein level for GLUT1 and GLUT8 (p < 0.05 and 0.01, respectively). The temporal expression pattern was shared with cytokeratin 18 and Ki67 (p < 0.01). Endogenous GLUT8 partially co-localized with 58 K protein and α-lactalbumin in mammary tissue and with Golgi membrane-associated protein 130 in isolated epithelial cells. The spatial-temporal synchrony between expression of GLUT8/GLUT1 and alveolar cell proliferation, and its localization in cis-Golgi associated to lactose synthase complex, suggest that both transporters are involved in glucose uptake into this organelle, supporting lactose synthesis.

Increased expression of mitochondrial sodium-coupled ascorbic acid transporter-2 (mitSVCT2) as a central feature in breast cancer.

The potential role of vitamin C in cancer prevention and treatment remains controversial. While normal human cells obtain vitamin C as ascorbic acid, the prevalent form of vitamin C in vivo, the uptake mechanisms by which cancer cells acquire vitamin C has remained unclear. The aim of this study is to characterize how breast cancer cells acquire vitamin C. For this, we determined the expression of vitamin C transporters in normal and breast cancer tissue samples, and in ZR-75, MCF-7, MDA-231 and MDA-468 breast cancer cell lines. At the same time, reduced (AA) and oxidized (DHA) forms of vitamin C uptake experiments were performed in all cell lines. We show here that human breast cancer tissues differentially express a form of SVCT2 transporter, that is systematically absent in normal breast tissues and it is increased in breast tumors. In fact, estrogen receptor negative breast cancer tissue, exhibit the most elevated SVCT2 expression levels. Despite this, our analysis in breast cancer cell lines showed that these cells are not able to uptake ascorbic acid and depend on glucose transporter for the acquisition of vitamin C by a bystander effect. This is consistent with our observations that this form of SVCT2 is completely absent from the plasma membrane and is overexpressed in mitochondria of breast cancer cells, where it mediates ascorbic acid transport. This work shows that breast cancer cells acquire vitamin C in its oxidized form and are capable of accumulated high concentrations of the reduced form. Augmented expression of an SVCT2 mitochondrial form appears to be a common hallmark across all human cancers and might have implications in cancer cells survival capacity against pro-oxidant environments.

Sustained blockade of ascorbic acid transport associated with marked SVCT1 loss in rat hepatocytes containing increased ascorbic acid levels after partial hepatectomy.

The liver has an extraordinary regenerative capacity in response to partial hepatectomy (PHx), which develops with neither tissue inflammation response nor alterations in the whole organism. This process is highly coordinated and it has been associated with changes in glutathione (GSH) metabolism. However, there are no reports indicating ascorbic acid (AA) levels after partial hepatectomy. AA and GSH act integrally as an antioxidant system that protects cells and tissues from oxidative damage and imbalance observed in a variety of diseases that affect the liver. Although rat hepatocytes are able to synthesize AA and GSH, which are the providers of AA for the whole organism, they also acquire AA from extracellular sources through the sodium-coupled ascorbic acid transporter-1 (SVCT1). Here, we show that hepatocytes from rat livers subjected to PHx increase their GSH and AA levels from 1 to 7 days post hepatectomy, whose peaks precede the peak in cell proliferation observed at 3 days post-hepatectomy. The increase in both antioxidants was associated with higher expression of the enzymes involved in their synthesis, such as the modifier subunit of enzyme glutamine cysteine ligase (GCLM), glutathione synthetase (GS), gulonolactonase (GLN) and gulonolactone oxidase (GULO). Importantly, rat hepatocytes, that normally exhibit kinetic evidence indicating only SVCT1-mediated transport of AA, lost more than 90% of their capacity to transport it at day 1 after PHx without evidence of recovery at day 7. This observation was in agreement with loss of SVCT1 protein expression, which was undetectable in hepatocytes as early as 2h after PHx, with partial recovery at day 7, when the regenerated liver weight returns to normal. We conclude that after PHx, rat hepatocytes enhance their antioxidant capacity by increasing GSH and AA levels prior to the proliferative peak. GSH and AA are increased by de novo synthesis, however paradoxically hepatocytes from rat subjected to PHx also suppress their capacity to acquire AA from extracellular sources through SVCT1.

B7-H4(B7x)-Mediated Cross-talk between Glioma-Initiating Cells and Macrophages via the IL6/JAK/STAT3 Pathway Lead to Poor Prognosis in Glioma Patients.

PURPOSE: The objective of this study was to evaluate clinical significance and immunosuppressive mechanisms of B7-H4 (B7x/B7S1), a B7 family member, in glioma. EXPERIMENTAL DESIGN: B7-H4 levels in glioma tissue/cerebral spinal fluid (CSF) were compared between different grades of glioma patients. Survival data were analyzed with Kaplan-Meier to determine the prognostic value of B7-H4. Cytokines from CD133(+) cells to stimulate the expression of B7-H4 on human macrophages (Mφs) were investigated by FACS, neutralizing antibodies, and Transwell chemotaxis assay. shRNA, reporter vector, and chromatin immunoprecipitation were used to determine the binding of STAT3 to the B7-H4 promoter. The function of B7-H4(+) Mφs in vitro was evaluated through phagocytosis, T-cell proliferation/apoptosis, and cytokine production as well as in the xenografted model for in vivo analysis. RESULTS: We found that B7-H4 expression in tumors was associated with prognosis of human glioblastoma and correlated directly with malignant grades. Mechanistically, glioma initiating CD133(+) cells and Mφs/microglia cointeraction activated expression of B7-H4 via IL6 and IL10 in both tumor cells and microenvironment supporting cells. IL6-activated STAT3 bound to the promoter of B7-H4 gene and enhanced B7-H4 expression. Furthermore, CD133(+) cells mediated immunosuppression through B7-H4 expression on Mφs/microglia by silencing of B7-H4 expression on these cells, which led to increased microenvironment T-cell function and tumor regression in the xenograft glioma mouse model. CONCLUSIONS: We have identified B7-H4 activation on Mφs/microglia in the microenvironment of gliomas as an important immunosuppressive event blocking effective T-cell immune responses. Clin Cancer Res; 22(11); 2778-90. ©2016 AACR.

PP2A inhibition with LB100 enhances cisplatin cytotoxicity and overcomes cisplatin resistance in medulloblastoma cells.

The protein phosphatase 2A (PP2A) inhibitor, LB100, has been shown in pre-clinical studies to be an effective chemo- and radio-sensitizer for treatment of various cancers. We investigated effects associated with LB100 treatment alone and in combination with cisplatin for medulloblastoma (MB) in vitro and in vivo in an intracranial xenograft model. We demonstrated that LB100 had a potent effect on MB cells. By itself, LB100 inhibited proliferation and induced significant apoptosis in a range of pediatric MB cell lines. It also attenuated MB cell migration, a pre-requirement for invasion. When used in combination, LB100 enhanced cisplatin-mediated cytotoxic effects. Cell viability in the presence of 1 uM cisplatin alone was 61% (DAOY), 100% (D341), and 58% (D283), but decreased with the addition of 2 µM of LB100 to 26% (DAOY), 67% (D341), and 27% (D283), (p < 0.005). LB100 suppressed phosphorylation of the STAT3 protein and several STAT3 downstream targets. Also, LB100 directly increased cisplatin uptake and overcame cisplatin-resistance in vitro. Finally, LB100 exhibited potent in vivo anti-neoplastic activity in combination with cisplatin in an intracranial xenograft model.

Photodynamic therapy (PDT) for malignant brain tumors--where do we stand?

INTRODUCTION: What is the current status of photodynamic therapy (PDT) with regard to treating malignant brain tumors? Despite several decades of effort, PDT has yet to achieve standard of care. PURPOSE: The questions we wish to answer are: where are we clinically with PDT, why is it not standard of care, and what is being done in clinical trials to get us there. METHOD: Rather than a meta-analysis or comprehensive review, our review focuses on who the major research groups are, what their approaches to the problem are, and how their results compare to standard of care. Secondary questions include what the effective depth of light penetration is, and how deep can we expect to kill tumor cells. CURRENT RESULTS: A measurable degree of necrosis is seen to a depth of about 5mm. Cavitary PDT with hematoporphyrin derivative (HpD) results are encouraging, but need an adequate Phase III trial. Talaporfin with cavitary light application appears promising, although only a small case series has been reported. Foscan for fluorescence guided resection (FGR) plus intraoperative cavitary PDT results were improved over controls, but are poor compared to other groups. 5-Aminolevulinic acid-FGR plus postop cavitary HpD PDT show improvement over controls, but the comparison to standard of care is still poor. CONCLUSION: Continued research in PDT will determine whether the advances shown will mitigate morbidity and mortality, but certainly the potential for this modality to revolutionize the treatment of brain tumors remains. The various uses for PDT in clinical practice should be pursued.

Duplicación de intestino delgado / Small bowel duplicity

Las duplicaciones intestinales son malformaciones congénitas poco frecuentes. Pueden localizarse desde la boca al ano. En general son asintomáticas, aunque pueden complicarse y manifestarse como hemorragia digestiva o abdomen agudo en aquellos casos en que se localiza a nivel abdominal. Esto obliga a tenerlo en cuenta como diagnóstico diferencial ante estos cuadros. En adultos, el diagnóstico suele darse de manera casual en el acto operatorio. Si bien la conducta es quirúrgica, con resección de la duplicación, ya sea total o parcial, en casos asintomáticos no complicados, esta conducta es controversial. Se presentan tres casos asintomáticos no complicados, diagnosticados casualmente en operaciones abdominales por otras causas, en la que se optó por tratamiento conservador. Dos de los casos fueron operados en el Hospital Regional del Luque y el tercero en el Instituto Nacional del Cáncer. Se hace una revisión del tema.

Intestinal duplications are infrequent congenital malformations. They can be located since the mouth to the anus. Generally they are asymptomatic, although they can be complicated and manifest themselves as a gastrointestinal bleeding or an acute abdomen in those cases that they are located in an abdominal level. These cases compel us to think in them as a differential diagnosis. In adults, the diagnosis usually occurs by chance during the surgical procedure. Although the behavior is surgical, with a partial or total resection of the duplication, it is controversial in those asymptomatic or not complicated cases. It is presented three asymptomatic and not complicated cases which were diagnosed by chance during abdominal surgeries for other causes, and where we chose a conservative treatment. Two out of the three cases were operated in the Hospital Regional de Luque and the third case was operated in the Instituto Nacional del Cáncer. It is made a review of the subject.

Sequential combination of decitabine and idarubicin synergistically enhances anti-leukemia effect followed by demethylating Wnt pathway inhibitor promoters and downregulating Wnt pathway nuclear target.

BACKGROUND: The methylation inhibitor 5-Aza-2'-deoxycytidine (decitabine, DAC) has a great therapeutic value for acute myeloid leukemia (AML) and myelodysplastic syndromes (MDS). But decitabine monotherapy was associated with a relatively low rate of complete remission in AML and MDS. We aimed to investigate the effect of several anti-leukemia drugs in combination with decitabine on the proliferation of myeloid leukemia cells, to select the most efficient combination group and explore the associated mechanisms of these combination therapies. METHODS: Cell proliferation was tested by MTT assay and CFU-GM assay. Cell apoptosis was evaluated by Annexin V and PI staining in cell culture, TUNEL assay and transmission electron microscopy in animal study. MicroPET was used to imaging the tumor in mouse model. Molecular studies were conducted using microarray expression analysis, which was used to explore associated pathways, and real-time quantitative reverse transcription-PCR, western blot and immunohistochemistry, used to assess regulation of Wnt/ß-catenin pathway. Statistical significance among groups was determined by one-way ANOVA analysis followed by post hoc Bonferroni's multiple comparison test. RESULTS: Among five anti-leukemia agents in combining with decitabine, the sequential combination of decitabine and idarubicin induced synergistic cell death in U937 cells, and this effect was verified in HEL, SKM-1 cells and AML cells isolated from AML patients. Importantly, tumor growth inhibition in this sequential combination was found to be higher than in single agent or controls in vivo. Moreover, sequential combination of the two agents induced apoptosis and depression of the Wnt/ß-catenin pathway in both AML cell culture and animal studies. CONCLUSIONS: The findings demonstrated that sequentially combination of decitabine and idarubicin had synergistic anti-leukemia effects. These effects were mainly attributed to demethylation of Wnt/ß-catenin pathway inhibitors and downregulation of Wnt/ß-catenin pathway nuclear targets.

Noncompetitive blocking of human GLUT1 hexose transporter by methylxanthines reveals an exofacial regulatory binding site.

Glucose transporter (GLUT)1 has become an attractive target to block glucose uptake in malignant cells since most cancer cells overexpress GLUT1 and are sensitive to glucose deprivation. Methylxanthines are natural compounds that inhibit glucose uptake; however, the mechanism of inhibition remains unknown. Here, we used a combination of binding and glucose transport kinetic assays to analyze in detail the effects of caffeine, pentoxifylline, and theophylline on hexose transport in human erythrocytes. The displacement of previously bound cytochalasin B revealed a direct interaction between the methylxanthines and GLUT1. Methylxanthines behave as noncompetitive blockers (inhibition constant values of 2-3 mM) in exchange and zero-trans efflux assays, whereas mixed inhibition with a notable uncompetitive component is observed in zero-trans influx assays (inhibition constant values of 5-12 mM). These results indicate that methylxanthines do not bind to either exofacial or endofacial d-glucose-binding sites but instead interact at a different site accessible by the external face of the transporter. Additionally, infinite-cis exit assays (Sen-Widdas assays) showed that only pentoxifylline disturbed d-glucose for binding to the exofacial substrate site. Interestingly, coinhibition assays showed that methylxanthines bind to a common site on the transporter. We concluded that there is a methylxanthine regulatory site on the external surface of the transporter, which is close but distinguishable from the d-glucose external site. Therefore, the methylxanthine moiety may become an attractive framework for the design of novel specific noncompetitive facilitative GLUT inhibitors.

Ensayo clínico de asignación aleatoria, para evaluar la eficacia de dos técnicas de compresión en la disminución de complicaciones en el sitio de acceso vascular femoral, posterior a cateterismo diagnóstico y terapéutico / Randomized, controlled trial, to assess the efficacy of two vascular compression techniques in reducing complications at vascular access sites after therapeutic or diagnostic catheterization

Objetivo: Comparar la incidencia de falla en la hemostasia y frecuencia de eventos vasculares, durante y después de la compresión con dos maniobras diferentes. Métodos: Se realizó un ensayo clínico de asignación aleatoria a compresión mecánica o compresión manual, para el retiro de introductor en arteria femoral posterior a cateterismo diagnóstico o terapéutico. Resultados: Se incluyeron 100 pacientes en el grupo de compresión con compresor (grupo uno) y 112 de forma manual (grupo dos). La falla para lograr hemostasia se presentó en 48% del grupo uno vs 19.7% en el grupo dos (p<0.001). Existió una tendencia a presentar con más frecuencia hematomas >4 cm durante el seguimiento con ultrasonido, en el grupo con compresor 11.4% vs el grupo manual 4.6% (p=0.062). En el análisis de regresión logística sólo se encontró que los factores independientes para falla en la hemostasia fueron: uso de compresor con OR 4.34 (IC 95%, 2.24-8.43, p<0.001) y edad mayor a 61 años con OR 2.44 (IC 95%, 1.3-4.7, p=0.008), el índice de masa corporal < 26 disminuyó el riesgo con OR 0.86 (IC 95%, 0.78-0.94, p=0.001). Conclusiones: Para el retiro de introductores, la compresión manual es superior al empleo del compresor mecánico para evitar la falla de hemostasia en el sitio de punción. Una limitación es que no se puede asegurar cuál de los dos métodos es superior para disminuir complicaciones vasculares, dado que no se completó el tamaño de muestra calculado.

Objective: To compare the incidence of hemostatic failure and rate of vascular events during and after vascular compression using two different techniques. Methods: Patients were randomized to mechanical or manual compression after a therapeutic or diagnostic catheterization procedure. Results: One hundred patients were enrolled in the mechanical compression group (group one) and 112 patients in the manual compression group (group two). Failed hemostasis was observed in 48% of patients in group 1 and 19.7% in group two (p<0.001). A tendency towards a greater incidence for hematoma (>4cm) formation was found on ultrasound follow-up in group 1 (11.4% vs 4.6%, p=0.062). Logistic regression analysis found that the only independent factors for hemostatic failure were: use of mechanical compression device (OR 4.34, 95% CI 2.24-8.43, p<0.001) and age greater than 61 years (OR 2.44, 95% CI 1.3-4.7, p=0.008). A body mass index <26 was found to reduce the risk for hemostatic failure (OR 0.84, 95% CI 0.78-0.94, p=0.001). Conclusion: After introducer sheath removal, manual compression is superior to mechanically-assisted hemostasis in avoiding hemostatic failure at vascular access site. This study was not able, however, to show the superiority of either method to reduce the rate of vascular complications due to the small sample size of patients enrolled at the time of early study termination for safety reasons.

Essential role of intracellular glutathione in controlling ascorbic acid transporter expression and function in rat hepatocytes and hepatoma cells.

Although there is in vivo evidence suggesting a role for glutathione in the metabolism and tissue distribution of vitamin C, no connection with the vitamin C transport systems has been reported. We show here that disruption of glutathione metabolism with buthionine-(S,R)-sulfoximine (BSO) produced a sustained blockade of ascorbic acid transport in rat hepatocytes and rat hepatoma cells. Rat hepatocytes expressed the Na(+)-coupled ascorbic acid transporter-1 (SVCT1), while hepatoma cells expressed the transporters SVCT1 and SVCT2. BSO-treated rat hepatoma cells showed a two order of magnitude decrease in SVCT1 and SVCT2 mRNA levels, undetectable SVCT1 and SVCT2 protein expression, and lacked the capacity to transport ascorbic acid, effects that were fully reversible on glutathione repletion. Interestingly, although SVCT1 mRNA levels remained unchanged in rat hepatocytes made glutathione deficient by in vivo BSO treatment, SVCT1 protein was absent from the plasma membrane and the cells lacked the capacity to transport ascorbic acid. The specificity of the BSO treatment was indicated by the finding that transport of oxidized vitamin C (dehydroascorbic acid) and glucose transporter expression were unaffected by BSO treatment. Moreover, glutathione depletion failed to affect ascorbic acid transport, and SVCT1 and SVCT2 expression in human hepatoma cells. Therefore, our data indicate an essential role for glutathione in controlling vitamin C metabolism in rat hepatocytes and rat hepatoma cells, two cell types capable of synthesizing ascorbic acid, by regulating the expression and subcellular localization of the transporters involved in the acquisition of ascorbic acid from extracellular sources, an effect not observed in human cells incapable of synthesizing ascorbic acid.

Histidine residues in the Na+-coupled ascorbic acid transporter-2 (SVCT2) are central regulators of SVCT2 function, modulating pH sensitivity, transporter kinetics, Na+ cooperativity, conformational stability, and subcellular localization.

Na(+)-coupled ascorbic acid transporter-2 (SVCT2) activity is impaired at acid pH, but little is known about the molecular determinants that define the transporter pH sensitivity. SVCT2 contains six histidine residues in its primary sequence, three of which are exofacial in the transporter secondary structure model. We used site-directed mutagenesis and treatment with diethylpyrocarbonate to identify histidine residues responsible for SVCT2 pH sensitivity. We conclude that five histidine residues, His(109), His(203), His(206), His(269), and His(413), are central regulators of SVCT2 function, participating to different degrees in modulating pH sensitivity, transporter kinetics, Na(+) cooperativity, conformational stability, and subcellular localization. Our results are compatible with a model in which (i) a single exofacial histidine residue, His(413), localized in the exofacial loop IV that connects transmembrane helices VII-VIII defines the pH sensitivity of SVCT2 through a mechanism involving a marked attenuation of the activation by Na(+) and loss of Na(+) cooperativity, which leads to a decreased V(max) without altering the transport K(m); (ii) exofacial histidine residues His(203), His(206), and His(413) may be involved in maintaining a functional interaction between exofacial loops II and IV and influence the general folding of the transporter; (iii) histidines 203, 206, 269, and 413 affect the transporter kinetics by modulating the apparent transport K(m); and (iv) histidine 109, localized at the center of transmembrane helix I, might be fundamental for the interaction of SVCT2 with the transported substrate ascorbic acid. Thus, histidine residues are central regulators of SVCT2 function.