How do I: Evaluate the safety and legitimacy of unproven cellular therapies?

BACKGROUND: Unproven cellular therapies are being offered to patients for a variety of conditions and diseases for which other treatments have failed. The use of untested cellular therapies is a worldwide problem. Practitioners (e.g., physicians, scientists, QA/QI facility managers, and policy advocates) are perhaps unaware of the risks involved with such therapies. Therefore, a critical need exists to bring attention to the potential limitations and adverse effects of these therapies to inform and limit misinformation. STUDY DESIGN AND METHODS: We describe the extent of the unproven cellular therapy problem through a search of scientific literature and social media coverage. We also describe the regulatory framework that can be used by the practitioner to review and evaluate both proven and unproven cellular therapies. RESULTS: We report on the current state of unproven cellular therapies across the globe. A workflow to facilitate an understanding of the regulatory processes involved in the approval of cellular therapies is provided as well as a list of warnings required by regulatory agencies on various products. It is hoped that this article will serve as a tool kit to educate the practitioner on navigating the field of unproven cellular therapy products. DISCUSSION: Increasing awareness of the issues associated with unproven therapies through education is important to help in reducing misinformation and risks to patients.

SVCT2 transporter expression is post-natally induced in cortical neurons and its function is regulated by its short isoform.

Different studies have demonstrated the importance of micronutrients, such as vitamins, for normal adult brain function and development. Vitamin C is not synthesized in the brain, but high levels are detected in this organ because of the existence of specific uptake mechanisms, which concentrate ascorbic acid from the bloodstream to the cerebrospinal fluid and then into neurons and glial cells. Two different isoforms of sodium-vitamin C cotransporters (SVCT1 and SVCT2) have been cloned. SVCT2 expression has been observed in the adult hippocampus and cortical neurons by in situ hybridization. In addition, the localization of SVCT2 in the rat fetal brain has been studied by immunohistochemistry and in situ hybridization, demonstrating that SVCT2 is highly expressed in the ventricular and subventricular areas of the brain cortex. However, there are currently no immunohistochemical data regarding SVCT2 expression and function in the post-natal brain. Therefore, we analyzed SVCT2 expression in the developing brain cortex of mice, and demonstrated an increase in SVCT2 mRNA in mice at 1-15 days of age. The expression of a short isoform, SVCT2sh, was also detected within the same period. SVCT2 expression was concentrated in neurons within the inner layer of the brain cortex. Both SVCT2 isoforms were coexpressed in N2a cells to obtain functional data. Fluorescence resonance energy transfer analysis revealed a molecular interaction between SVCT2wt and SVCT2sh. Finally, differences in transport ratios suggested that SVCT2sh expression inhibited ascorbic acid uptake in N2a cells when both isoforms were coexpressed. The sodium-vitamin C cotransporter, SVCT2, is induced in neurons within the inner layer of the brain cortex during post-natal development, mainly in pyramidal cortex neurons. Two different isoforms, SVCT2wt and SVCT2sh, were detected. Using in vitro studies, we suggest a molecular interaction between SVCT2wt and SVCT2sh, which may regulate the affinity of vitamin C uptake.

Micropapillary carcinoma of the urinary tract: a cytologic study of urine and fine-needle aspirate samples.

OBJECTIVE: Micropapillary carcinoma (MPC) is an aggressive variant of urothelial carcinoma that needs early and specific recognition. In order to determine whether this tumor variant can be recognized with cytology, we evaluated a large cytohistological series. STUDY DESIGN: It was a retrospective cytohistological correlation study including 20 patients with MPC. Only those cases in which the tumor exhibited >50% of micropapillary growth were selected. Twenty exfoliative urine specimens and four needle aspirates from lymph node metastases were reviewed. RESULTS: On histology, 14 cases were infiltrative, while 6 were exclusively superficial. Cytology was characterized by numerous small, cohesive groups and single neoplastic cells. Pseudopapillae were present in 17 cases and in 9 they were a relevant finding. Morules were present in 15 cases. Isolated microacini were seen in 14 cases. Infiltrative tumors showed more neoplastic groups. Cellular atypia was prominent in 17 cases. In 15 cases, a cytologic diagnosis of urothelial carcinoma was made. One case was diagnosed as adenocarcinoma. The remaining 4 cases were considered suspicious of malignancy. CONCLUSIONS: The peculiar morphology of MPC of the urinary tract is partially reflected on cytology, allowing in some cases a specific recognition. This is important since the aggressive behavior of this neoplasm needs rapid management and treatment.

Contextual Factors-Enriched Standard Care on mechanical neck pain (ContextualizAR trial): Protocol for a randomised controlled trial.

BACKGROUND: Understanding the influence of contextual factors (CFs) on interventions for mechanical neck pain (MNP) is essential for evidence-based practice in physical therapy. However, the specific effects and synergies of combining different CFs remain unclear. OBJECTIVE: The primary purpose of this study will be to determine if a CFs-Enriched Standard Care (SC) approach is an effective treatment for MNP in terms of reducing pain and improving function. METHODS: This will be an assessor-blinded, 2-group (1:1) randomised clinical trial (RCT) aiming to enrol 94 participants with neck pain persisting for more than 4 weeks. Both groups will undergo 4 weeks of SC twice weekly, following established clinical practice guidelines. In the intervention group, CFs will be enhanced, encompassing the physical, psychological, and social elements inherent in the clinical encounter, based on existing evidence. The primary outcomes will encompass changes in pain and disability after 4 weeks of treatment, with a follow-up reassessment at week 12 post-treatment. Secondary outcomes will include changes in Active Range of Motion, Global Rating of Change, and Satisfaction with treatment. The change between groups after treatment and at the 12-week follow-up will be reported for all outcomes, considering the difference from scores recorded at baseline. RESULTS: We hypothesise that a 4-week CFs-Enriched SC approach will be superior to SC alone in terms of patient-reported disability and pain, with measurements conducted using the Northwick Park Neck Pain Questionnaire and the Numeric Pain Rating Scale, respectively. CONCLUSION: This RCT rigorously assesses the effect of purposeful manipulation of CFs during MNP treatment. By elucidating the role of these factors, our findings have the potential to significantly refine clinical practice in managing MNP, thereby enhancing patient care, and advancing the fields of physical therapy and rehabilitation.

Superoxide-dependent uptake of vitamin C in human glioma cells.

Glioblastomas are lethal brain tumors that resist current cytostatic therapies. Vitamin C may antagonize the effects of reactive oxygen species (ROS) generating therapies; however, it is often used to reduce therapy-related side effects despite its effects on therapy or tumor growth. Because the mechanisms of vitamin C uptake in gliomas are currently unknown, we evaluated the expression of the sodium-vitamin C cotransporter (SVCT) and facilitative hexose transporter (GLUT) families in human glioma cells. In addition, as microglial cells can greatly infiltrate high-grade gliomas (constituting up to 45% of cells in glioblastomas), the effect of TC620 glioma cell interactions with microglial-like HL60 cells on vitamin C uptake (Bystander effect) was determined. Although glioma cells expressed high levels of the SVCT isoform-2 (SVCT2), low functional activity, intracellular localization and the expression of the dominant-negative isoform (dnSVCT2) were observed. The increased glucose metabolic activity of glioma cells was evident by the high 2-Deoxy-d-glucose and dehydroascorbic acid (DHA) uptake rates through the GLUT isoform-1 (GLUT1), the main DHA transporter in glioblastoma. Co-culture of glioma cells and activated microglial-like HL60 cells resulted in extracellular ascorbic acid oxidation and high DHA uptake by glioma cells. This Bystander effect may explain the high antioxidative potential observed in high-grade gliomas. This study strongly suggests that the Bystander effect, that is, glioma cell interaction with oxidant-producing microglia, could be an important mechanism for glioma vitamin C loading in the absence of functional sodium-vitamin C cotransporter 2 (SVCT2) expression. The high cellular vitamin C load in glioma cells results from a high uptake of extracellular dehydroascorbic acid (DHA) generated by neighboring microglia. This Bystander effect may explain the high antioxidative potential observed in high-grade gliomas, considering that high-grade gliomas may be the only neoplasm where oxidant-producing microglia can almost equal the number of tumor cells.

Clinical and experimental approaches to knee cartilage lesion repair and mesenchymal stem cell chondrocyte differentiation.

Cartilage has poor regeneration capacity due to the scarcity of endogenous stem cells, its low metabolic activity and the avascular environment. Repair strategies vary widely, including microfracture, autologous or allogenic tissue implantation, and in vitro engineered tissues of autologous origin. However, unlike the advances that have been made over more than two decades with more complex organs, including vascular, cardiac or bone tissues, similar advances in tissue engineering for cartilage repair are lacking. Although the inherent characteristics of cartilage tissue, such as the lack of vascularity and low cellular diversity, suggest that it would be one of the more simple tissues to be engineered, its functional weight-bearing role and implant viability and adaptation make this type of repair more complex. Over the last decade several therapeutic approaches and innovative techniques show promise for lasting and functional regeneration of hyaline cartilage. Here we will analyze the main strategies for cartilage regeneration and discuss our experience.

Fine-needle aspiration cytology of mammary myofibroblastoma: a report of six cases.

OBJECTIVE: To evaluate cytologic features of mammary myofibroblastoma in order to establish the possibility of precise preoperative recognition. STUDY DESIGN: This was a multi-institutional study of 6 patients with myofibroblastoma (5 men and 1 woman) in which preoperatively fine-needle aspiration cytology was performed. Four cases showed classical histologic features, 1 corresponded to the cellular variant and the remaining 1 to the fibrous form. RESULTS: Except for 1 case, smears were cellular and distributed as irregular aggregates and single cells. Most groups showed a small amount of metachromatic stroma and capillaries. Cells retained cytoplasm and showed a spindle-to-plump oval morphology with moderate pleomorphism. Nuclear pleomorphism was present and was relevant in 1 case. Intranuclear pseudoinclusions and mast cells were present in 3 cases. No epithelial clusters were seen. Due to hypercellularity and pleomorphism 1 case was considered as suspicious for malignancy. The remaining 5 were diagnosed as low-grade mesenchymal lesions, and myofibroblastoma was suggested in 3. CONCLUSION: Cytologic features of myofibroblastoma reflect what is seen on histology. When such findings are correlated with image studies, preoperative recognition can be possible. This is especially true for male patients in whom the tumor is relatively frequent.

MACSima imaging cyclic staining (MICS) technology reveals combinatorial target pairs for CAR T cell treatment of solid tumors.

Many critical advances in research utilize techniques that combine high-resolution with high-content characterization at the single cell level. We introduce the MICS (MACSima Imaging Cyclic Staining) technology, which enables the immunofluorescent imaging of hundreds of protein targets across a single specimen at subcellular resolution. MICS is based on cycles of staining, imaging, and erasure, using photobleaching of fluorescent labels of recombinant antibodies (REAfinity Antibodies), or release of antibodies (REAlease Antibodies) or their labels (REAdye_lease Antibodies). Multimarker analysis can identify potential targets for immune therapy against solid tumors. With MICS we analysed human glioblastoma, ovarian and pancreatic carcinoma, and 16 healthy tissues, identifying the pair EPCAM/THY1 as a potential target for chimeric antigen receptor (CAR) T cell therapy for ovarian carcinoma. Using an Adapter CAR T cell approach, we show selective killing of cells only if both markers are expressed. MICS represents a new high-content microscopy methodology widely applicable for personalized medicine.

POCUS and POCDUS: essential tools for the evaluation and management of carotid artery pseudoaneurysms after a gunshot wound.

BACKGROUND: Evaluation of asymptomatic penetrating vascular injuries can be done with Point-of-care ultrasound (POCUS) and Point-of-care Doppler ultrasound (POCDUS). CASE PRESENTATION: A 21-year-old woman was admitted to the Emergency Department with a small wound and pain on the left side of her neck. The patient stated she was standing outside her home and suddenly felt acute pain in the neck. She denied trauma or being assaulted and reported no significant past medical or surgical history. On physical exam the only positive finding was a small gunshot entry wound on the left side of her neck without hard signs of vascular injury. Bedside POCUS demonstrated soft tissue swelling and a hematoma next to the left carotid artery. A round in shape bullet was visualized in contact with the posterior left common carotid artery wall and two small saccular pseudoaneurysms were seen at left common carotid artery wall. POCDUS showed a patent left carotid artery and turbulent flow in the two saccular aneurysms. A computed tomography angiogram (CTA) was performed confirming the findings and a stent in left carotid artery was placed. The patient tolerated the procedure well and was discharged 4 days after the procedure. At the sixth month follow-up, Doppler ultrasound showed patent stent and resolution of the muscular hematoma. CONCLUSIONS: Penetrating trauma-related vascular injuries are complex cases to handle within an acute setting. POCUS and POCDUS are increasingly being used for the workup and decision-making process of gunshot-related vascular injuries to the neck and are a fundamental part of the follow-up after definitive therapy.

Evaluación de la movilidad de la dorsiflexión de tobillo en cadena cinemática cerrada / Assessment of ankle dorsiflexion range of motion in closed kinetic chain

RESUMEN Los esguinces laterales de tobillo (ELT) son las lesiones musculoesqueléticas más prevalentes entre sujetos que participan de actividades físicas y deportes. Además, hasta un 70% de la población general ha sufrido un ELT, el cual presenta la mayor tasa de reincidencia de todas las afecciones musculoesqueléticas de los miembros inferiores. Luego de un ELT, se ha registrado una limitación de la dorsiflexión de tobillo en cadena cinemática cerrada (CCC), movimiento representativo en tareas funcionales e influyente en el retorno al deporte. La prueba de estocada con carga de peso es utilizada para evaluar la dorsiflexión de tobillo en CCC y existen diferentes formas de realizarla. El objetivo de este paso a paso es describir dos formas de medición de la dorsiflexión de tobillo en CCC mediante el uso de aplicaciones móviles.

ABSTRACT Lateral ankle sprains (LAS) are the most prevalent musculoskeletal injuries among subjects participating in physical activities and sports. In addition, up to 70% of the general population has sustained a LAS, with the highest rate of recurrence of all musculoskeletal injuries in the lower limbs. Subjects with a history of LAS have limited ankle dorsiflexion range of motion in closed kinetic chain (CKC), which is a common movement during functional tasks and an essential movement in return to sports. The weight-bearing lunge test is used to assess ankle dorsiflexion range of motion in CKC and may be performed through different procedures. The objective of this study is to describe two procedures for measuring ankle dorsiflexion in CKC through the use of mobile applications.

The FAST D protocol: a simple method to rule out traumatic vascular injuries of the lower extremities.

BACKGROUND: The aim of this study is to assess the accuracy of a Fast Doppler protocol for the examination of an injured lower limb, namely 2-Point Fast Doppler (2PFD), in order to rapidly triage arterial lesions after penetrating trauma. METHODS: The presence of flow and the aspects of the Doppler waveform of the dorsalis pedis artery (DPA) and posterior tibial artery (PTA) of the injured lower limb (2PFD) were evaluated immediately before the execution of a standardized Color Duplex Doppler (SD) evaluation in 149 limbs of 140 patients with gunshot penetrating injuries. We considered 2PFD normal exams as the ones with triphasic patterns in both the DPA and PTA, and 2PFD pathologic exams as the ones with absent, biphasic, or monophasic flow patterns in the DPA and/or PTA. 2PFD data were then analyzed to assess accuracy variables, using SD results as matching test reference. According to the trauma center standard protocols, SD positive cases underwent also angiography and surgical exploration, whose findings were used to further match the 2PFD specificity. RESULTS: The 2PFD protocol showed a sensitivity of 100%, and a specificity of 100% compared with the SD, in the diagnostic workup of arterial injuries of the lower limbs after penetrating trauma. Furthermore, all the pathologic cases that resulted in all true positives (TP), compared with SD, were confirmed as TP also when matched with the angiography evaluation results. CONCLUSIONS: The 2PFD protocol can rapidly identify arterial flow and differentiate between normal and pathologic spectral Doppler analyses in distal arteries. The presence of the normal triphasic flows in DPA and PTA is as sensitive as the standardized Color Doppler Duplex assessment of the entire limb in ruling out arterial lesions in lower-limb penetrating trauma. The absence of flow or the presence of a biphasic or monophasic pathologic flow in DPA and PTA is pathologic and should be always followed by further investigation. 2PFD is faster and easier to perform compared with the SD approach. It could become a new first-line screening technique, both in pre-hospital and hospital critical scenarios, particularly in contexts where advanced diagnostic performance is limited by time concerns or scarce resources.

Phosphate Starvation-Dependent Iron Mobilization Induces CLE14 Expression to Trigger Root Meristem Differentiation through CLV2/PEPR2 Signaling.

Low inorganic phosphate (Pi) availability causes terminal differentiation of the root apical meristem (RAM), a phenomenon known as root meristem exhaustion or determined growth. Here, we report that the CLE14 peptide acts as a key player in this process. Low Pi stress induces iron mobilization in the RAM through the action of LPR1/LPR2, causing expression of CLE14 in the proximal meristem region. CLV2 and PEPR2 receptors perceive CLE14 and trigger RAM differentiation, with concomitant downregulation of SHR/SCR and PIN/AUXIN pathway. Our results reveal multiple steps of the molecular mechanism of one of the most physiologically important root nutrient responses.

Búsqueda etiológica de ACV en salas de medicina del Hospital Pasteur, Montevideo - Uruguay. Julio - setiembre 2019 / Aetiological search for stroke in medical rooms of the Pasteur Hospital, Montevideo - Uruguay. July - September 2019 / Pesquisa etiológica da ACV em salas médicas do Hospital Pasteur, Montevideo - Uruguai. Julho - setembro 2019

Resumen: Introducción: El ataque cerebrovascular (ACV) es una lesión del parénquima cerebral por compromiso vascular, pudiendo ser hemorrágico o isquémico. Presenta alta morbimortalidad, siendo la segunda causa de muerte a nivel mundial, tercera en nuestro país, que genera además elevados costos sanitarios. Conocer las etiologías más frecuentes de esta patología, su asociación con factores de riesgo y el tratamiento realizado, podría contribuir a la correcta elaboración de estrategias de prevención primaria y secundaria. Objetivos: Conocer la/s etiología/s del ACV en pacientes internados en salas de medicina interna del Hospital Pasteur, sus características epidemiológicas; la asociación entre los factores de riesgo, paraclínica solicitada y tratamiento sugerido al alta. Metodología: Estudio descriptivo, transversal, observacional. Se incluyeron pacientes mayores de 18 años con diagnóstico de ACV internados en las salas de medicina del Hospital Pasteur, entre el 15 de Julio y el 30 de Septiembre del 2019. Se realizó la recolección de datos a partir de las historias clínicas por medio de un formulario precodificado, que analizó variables categóricas como factores de riesgo, presentación clínica, paraclínica solicitada y tratamiento. Resultados y discusión: Se incluyeron 30 pacientes, 18 de sexo femenino. La media de edad fue 70,48 años. La naturaleza isquémica predominó en un 90%. Los principales factores de riesgo fueron hipertensión arterial, dislipemia, diabetes mellitus y tabaquismo. El síndrome piramidal fue la forma de presentación clínica más frecuente. La etiología más prevalente fue de causa indeterminada, seguida por la ateroesclerótica y cardioembólica. En algunos pacientes no se realizó el algoritmo diagnóstico completo. El tratamiento realizado fue el adecuado de acuerdo a las causas planteadas. Conclusiones: La etiología más frecuente fue de causa indeterminada, seguida por la aterotrombótica. Los factores de riesgos más prevalentes fueron la hipertensión arterial, la dislipemia, la diabetes mellitus y el tabaquismo. La mayoría de los pacientes tenían más de 3 factores de riesgo asociados, de esta forma concluimos que la sumatoria de factores de riesgo incrementa la incidencia de ACV.

Abstract: Introduction: A stroke is a parenchymal cerebral lesion due to vascular compromise. This could be either hemorrhagic or ischemic. It presents high morbidity and mortality, being the second cause of death in the world and third in Uruguay, causing elevated health expenses. Recognizing the most frequent etiologies, their association with different risks factors and treatments applied could help to create better primary and secondary prevention strategies. Objectives: recognized stroke`s etiologies in patients hospitalized in Hospital Pasteur`s rooms, their epidemiologic characteristics, risk factors associated, the requested paraclinic and the treatment before they leave the hospital. Methods: Descriptive, observational and transversal study. We included patients older than 18 years old, with a stroke diagnosis, admitted into Hospital Pasteur medical`s rooms, between July 15 and September 30, 2019. Data were collected from medical records by accepting an informed consent, which analyzed variables such as risk factors, clinical presentation, requested preclinical and treatment. Results and discussion: 30 patients were included, 18 were female. The average age was 70,48 years. Ischemic strokes were predominated by 90%. Hypertension, dyslipidemia, diabetes mellitus and smoking were the predominant risk factors. Pyramidal syndrome was the most frequent clinical presentation. Undetermined stroke was the principal etiology, followed by atherosclerotic and cardioembolic. There were some patients that the algorithm was not completed. Treatments were adequate according to the causes. Conclusions: The most frequent etiology was the undetermined cause, followed by atherothrombotic. Prevalent risk factors were hypertension, dyslipidemia, DM and smoking. Most of the patients had more than 3 associated risk factors, so we conclude that the sum of risk factors increases the incidence of stroke.

Resumo: Introdução: O ataque cerebrovascular (AVC) é uma lesão do parênquima cerebral por comprometimento vascular, podendo ser hemorrágico ou isquêmico. Apresenta alta morbimortalidade, sendo a segunda principal causa de morte no mundo, a terceira em nosso país, o que também gera altos custos de saúde. Conhecer as etiologias mais frequentes dessa patologia, sua associação com fatores de risco e o tratamento realizado, poderia contribuir para o correto desenvolvimento de estratégias de prevenção primária e secundária. Objetivos: Conhecer a etiologia / s do AVC em pacientes internados nas salas de medicina interna do Hospital Pasteur, suas características epidemiológicas; a associação entre fatores de risco, solicitou tratamento paraclínico e sugerido no momento da alta. Metodologia: Estudo descritivo, transversal, observacional. Entre 15 de julho e 30 de setembro de 2019, foram incluídos pacientes com idade superior a 18 anos com diagnóstico de AVC admitidos nas salas médicas do Hospital Pasteur. Os dados foram coletados dos prontuários por meio de formulário pré-codificado, que analisou variáveis ​​categóricas como fatores de risco, apresentação clínica, solicitação paraclínica e tratamento. Resultados e discussão: foram incluídos 30 pacientes, 18 do sexo feminino. A idade média foi de 70,48 anos. A natureza isquêmica predominou em 90%. Os principais fatores de risco foram pressão alta, dislipidemia, diabetes mellitus e tabagismo. A síndrome piramidal foi a apresentação clínica mais frequente. A etiologia mais prevalente foi de causa indeterminada, seguida de aterosclerótica e cardioembólica. Em alguns pacientes, o algoritmo diagnóstico completo não foi realizado. O tratamento foi adequado de acordo com as causas. Conclusões: A etiologia mais frequente foi de causa indeterminada, seguida de aterotrombótica. Os fatores de risco mais prevalentes foram hipertensão, dislipidemia, DM e tabagismo. A maioria dos pacientes apresentou mais de três fatores de risco associados, concluindo que a soma dos fatores de risco aumenta a incidência de acidente vascular cerebral.

Identification of a new Alcaligenes faecalis strain MOR02 and assessment of its toxicity and pathogenicity to insects.

We report the isolation of a bacterium from Galleria mellonella larva and its identification using genome sequencing and phylogenomic analysis. This bacterium was named Alcaligenes faecalis strain MOR02. Microscopic analyses revealed that the bacteria are located in the esophagus and intestine of the nematodes Steinernema feltiae, S. carpocapsae, and H. bacteriophora. Using G. mellonella larvae as a model, when the larvae were injected with 24,000 CFU in their hemocoel, more than 96% mortality was achieved after 24 h. Additionally, toxicity assays determined that 1 µg of supernatant extract from A. faecalis MOR02 killed more than 70% G. mellonella larvae 96 h after injection. A correlation of experimental data with sequence genome analyses was also performed. We discovered genes that encode proteins and enzymes that are related to pathogenicity, toxicity, and host/environment interactions that may be responsible for the observed phenotypic characteristics. Our data demonstrates that the bacteria are able to use different strategies to colonize nematodes and kill insects to their own benefit. However, there remains an extensive group of unidentified microorganisms that could be participating in the infection process. Additionally, a nematode-bacterium association could be established probably as a strategy of dispersion and colonization.

Differential regulation of glucose transporter expression by estrogen and progesterone in Ishikawa endometrial cancer cells.

Estrogen replacement therapy and other unopposed estrogen treatments increase the incidence of endometrial abnormalities, including cancer. However, this effect is counteracted by the co-administration of progesterone. In the endometrium, glucose transporter (GLUT) expression and glucose transport are known to fluctuate throughout the menstrual cycle. Here, we determined the effect of estrogen and progesterone on the expression of GLUT1-4 and on the transport of deoxyglucose in Ishikawa endometrial cancer cells. Cells were incubated with estrogen, progesterone or combined estrogen and progesterone for 24 h and the effect on the expression of GLUT1-4 and on deoxyglucose transport was determined. We show that GLUT1 expression is upregulated by estrogen and progesterone individually, but that combined estrogen and progesterone treatment reverses this increase. Hormonal treatments do not affect GLUT2, GLUT3 or GLUT4 expression. Transport studies demonstrate that estrogen increases deoxyglucose transport at Michaelis-Menten constants (Kms) corresponding to GLUT1/4, an effect which disappears when progesterone is added concomitantly. These data demonstrate that different hormonal treatments differentially regulate GLUT expression and glucose transport in this endometrial cancer cell line. This regulation mirrors the role played by estrogen and progesterone on the incidence of cancer in this tissue and suggests that GLUT1 may be utilized by endometrial cancer cells to fuel their demand for increased energy requirement.

Response to combination therapy of interferon alfa-2b plus ribavirin in Hispanics with chronic hepatitis C.

OBJECTIVE: To evaluate the response rate of Hispanics with chronic hepatitis C to combination therapy of interferon alfa-2b plus ribavirin and to assess its adverse events. BACKGROUND: Hepatitis C virus may lead to chronic infection and multiple complications. Response to combination therapy of interferon plus ribavirin has been studied in many populations. African Americans have been found to have a lower response rate than Caucasians. However, little data exist for Hispanics. METHODS: Hispanic patients from Puerto Rico with chronic hepatitis C were eligible for the study between November 1997 and February 2000. The Institutional Review Boards of the participating institutions approved the study. Written informed consents were obtained. Combination therapy was given for 48 weeks and patients were followed for 24 weeks after treatment. Analysis of response to therapy was performed in an intention-to-treat basis. RESULTS: The most frequent adverse event was anemia (89%), associated to ribavirin. Sustained response was 23% for naive patients, 45% for relapsers, and 8% for non-responders to previous interferon monotherapy (p < 0.001). Data to analyze response was not available in 27% of patients. Hispanic patients had a low response rate to combination therapy. CONCLUSIONS: Response rates to combination therapy for Hispanic naive and previously non-responder patients are lower than in other reported populations. This may be due to a high prevalence of genotype 1 in Puerto Rico, which is associated to poor response. The higher response rate of relapsers, similar to those reported previously, was expected since these patients showed a previous response to interferon monotherapy. Ethnic factors may play a role in the response to therapy and should be further studied to determine proper treatment strategies for this population.

Lectin-like oxidized LDL receptor-1 is an enhancer of tumor angiogenesis in human prostate cancer cells.

Altered expression and function of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1) has been associated with several diseases such as endothelial dysfunction, atherosclerosis and obesity. In these pathologies, oxLDL/LOX-1 activates signaling pathways that promote cell proliferation, cell motility and angiogenesis. Recent studies have indicated that olr1 mRNA is over-expressed in stage III and IV of human prostatic adenocarcinomas. However, the function of LOX-1 in prostate cancer angiogenesis remains to be determined. Our aim was to analyze the contribution of oxLDL and LOX-1 to tumor angiogenesis using C4-2 prostate cancer cells. We analyzed the expression of pro-angiogenic molecules and angiogenesis on prostate cancer tumor xenografts, using prostate cancer cell models with overexpression or knockdown of LOX-1 receptor. Our results demonstrate that the activation of LOX-1 using oxLDL increases cell proliferation, and the expression of the pro-angiogenic molecules VEGF, MMP-2, and MMP-9 in a dose-dependent manner. Noticeably, these effects were prevented in the C4-2 prostate cancer model when LOX-1 expression was knocked down. The angiogenic effect of LOX-1 activated with oxLDL was further demonstrated using the aortic ring assay and the xenograft model of tumor growth on chorioallantoic membrane of chicken embryos. Consequently, we propose that LOX-1 activation by oxLDL is an important event that enhances tumor angiogenesis in human prostate cancer cells.

Characterizing comorbidities, a step to organize the patient's problems list / Caracterizando comorbidades, um passo para organizar a lista de problemas

Uma lista de problemas precisa e atualizada é essencial em um Registro Eletrônico em Saúde (RES) orientada a problemas. A falta de organização e categorização dos problemas limitam o resultado. Há certos problemas que afetam mais a evolução clínica do paciente, o que denominamos comorbidades. OBJETIVOS: O propósito deste artigo é caracterizar o conceito de comorbidade; bem como, classificar e identificar as comorbidades. MÉTODOS: Realizamos uma busca bibliográfica para caracterizar o conceito de comorbilidade. Posteriormente analisamos a RES do Hospital de Buenos Aires desde 1998 a 2015. Todos os problemas registrados nas epícrises como comorbidades foram tomados para análise. RESULTADOS: 20.849 ID-conceitos foram registrados e identificados inicialmente na epícrises. 90% foi levado para análise individual. 614 conceitos foram identificados como comorbidades (80,24% de registros). CONCLUSÕES: A maioria dos problemas analisados foram caracterizados como comorbidades, condizendo com a observação que gerou uma lista com as comorbidades mais frequentes.(AU)

An accurate and updated problems list is critical in a problem oriented Electronic Health Record (EHR). Thelack of organization and categorization of the problems limits the value of the list. There are certain problems that affect more than others the clinical evolution of the patient, these are known as comorbidities. OBJECTIVS: Characterize the comorbidity concept and identify, and classify the comorbidities. METHODS: A literature search of available definitions to characterize the concept "comorbidity". Then we analyzed theEHR in a Hospital from 1998 to 2015. All recorded problems as comorbidities conditions in patient discharge summaries were taken for analysis. RESULTS: 20,849 ID-concepts were initially obtained as comorbidities in patient discharge summaries. The 90% more frequent were taken for individual analysis. 614 ID-concepts were identied as comorbidities (80.24% of all records). CONCLUSIONS: Most of the problems analyzed were characterized as comorbidities making a list of the most frequently recorded.(AU)

Glial glucokinase expression in adult and post-natal development of the hypothalamic region.

It has recently been proposed that hypothalamic glial cells sense glucose levels and release lactate as a signal to activate adjacent neurons. GK (glucokinase), the hexokinase involved in glucose sensing in pancreatic beta-cells, is also expressed in the hypothalamus. However, it has not been clearly determined if glial and/or neuronal cells express this protein. Interestingly, tanycytes, the glia that cover the ventricular walls of the hypothalamus, are in contact with CSF (cerebrospinal fluid), the capillaries of the arcuate nucleus and adjacent neurons; this would be expected for a system that can detect and communicate changes in glucose concentration. Here, we demonstrated by Western-blot analysis, QRT-PCR [quantitative RT-PCR (reverse transcription-PCR)] and in situ hybridization that GK is expressed in tanycytes. Confocal microscopy and immuno-ultrastructural analysis revealed that GK is localized in the nucleus and cytoplasm of beta1-tanycytes. Furthermore, GK expression increased in these cells during the second week of post-natal development. Based on this evidence, we propose that tanycytes mediate, at least in part, the mechanism by which the hypothalamus detects changes in glucose concentrations.