Does HLA explain the high incidence of childhood-onset type 1 diabetes in the Canary Islands? The role of Asp57 DQB1 molecules.

The Canary Islands inhabitants, a recently admixed population with significant North African genetic influence, has the highest incidence of childhood-onset type 1 diabetes (T1D) in Spain and one of the highest in Europe. HLA accounts for half of the genetic risk of T1D. AIMS: To characterize the classical HLA-DRB1 and HLA-DQB1 alleles in children from Gran Canaria with and without T1D. METHODS: We analyzed classic HLA-DRB1 and HLA-DQB1 alleles in childhood-onset T1D patients (n = 309) and control children without T1D (n = 222) from the island of Gran Canaria. We also analyzed the presence or absence of aspartic acid at position 57 in the HLA-DQB1 gene and arginine at position 52 in the HLA-DQA1 gene. Genotyping of classical HLA-DQB1 and HLA-DRB1 alleles was performed at two-digit resolution using Luminex technology. The chi-square test (or Fisher's exact test) and odds ratio (OR) were computed to assess differences in allele and genotype frequencies between patients and controls. Logistic regression analysis was also used. RESULTS: Mean age at diagnosis of T1D was 7.4 ± 3.6 years (46% female). Mean age of the controls was 7.6 ± 1.1 years (55% female). DRB1*03 (OR = 4.2; p = 2.13-13), DRB1*04 (OR = 6.6; p ≤ 2.00-16), DRB1* 07 (OR = 0.37; p = 9.73-06), DRB1*11 (OR = 0.17; p = 6.72-09), DRB1*12, DRB1*13 (OR = 0.38; p = 1.21-05), DRB1*14 (OR = 0.0; p = 0.0024), DRB1*15 (OR = 0.13; p = 7.78-07) and DRB1*16 (OR = 0.21; p = 0.003) exhibited significant differences in frequency between groups. Among the DQB1* alleles, DQB1*02 (OR: 2.3; p = 5.13-06), DQB1*03 (OR = 1.7; p = 1.89-03), DQB1*05 (OR = 0.64; p = 0.027) and DQB1*06 (OR = 0.19; p = 6.25-14) exhibited significant differences. A total of 58% of the studied HLA-DQB1 genes in our control population lacked aspartic acid at position 57. CONCLUSIONS: In this population, the overall distributions of the HLA-DRB1 and HLA-DQB1 alleles are similar to those in other European populations. However, the frequency of the non-Asp-57 HLA-DQB1 molecules is greater than that in other populations with a lower incidence of T1D. Based on genetic, historical and epidemiological data, we propose that a common genetic background might help explain the elevated pediatric T1D incidence in the Canary Islands, North-Africa and middle eastern countries.

Characterization of Partial Discharges in Dielectric Oils Using High-Resolution CMOS Image Sensor and Convolutional Neural Networks.

In this work, an exhaustive analysis of the partial discharges that originate in the bubbles present in dielectric mineral oils is carried out. To achieve this, a low-cost, high-resolution CMOS image sensor is used. Partial discharge measurements using that image sensor are validated by a standard electrical detection system that uses a discharge capacitor. In order to accurately identify the images corresponding to partial discharges, a convolutional neural network is trained using a large set of images captured by the image sensor. An image classification model is also developed using deep learning with a convolutional network based on a TensorFlow and Keras model. The classification results of the experiments show that the accuracy achieved by our model is around 95% on the validation set and 82% on the test set. As a result of this work, a non-destructive diagnosis method has been developed that is based on the use of an image sensor and the design of a convolutional neural network. This approach allows us to obtain information about the state of mineral oils before breakdown occurs, providing a valuable tool for the evaluation and maintenance of these dielectric oils.

Applications of Modified Mesenchymal Stem Cells as Targeted Systems against Tumor Cells.

Combined gene and cell therapy are promising strategies for cancer treatment. Given the complexity of cancer, several approaches are actively studied to fight this disease. Using mesenchymal stem cells (MSCs) has demonstrated dual antitumor and protumor effects as they exert massive immune/regulatory effects on the tissue microenvironment. MSCs have been widely investigated to exploit their antitumor target delivery system. They can be genetically modified to overexpress genes and selectively or more efficiently eliminate tumor cells. Current approaches tend to produce more effective and safer therapies using MSCs or derivatives; however, the effect achieved by engineered MSCs in solid tumors is still limited and depends on several factors such as the cell source, transgene, and tumor target. This review describes the progress of gene and cell therapy focused on MSCs as a cornerstone against solid tumors, addressing the different MSC-engineering methods that have been approached over decades of research. Furthermore, we summarize the main objectives of engineered MSCs against the most common cancers and discuss the challenges, limitations, risks, and advantages of targeted treatments combined with conventional ones.

shRNA-Targeting Caspase-3 Inhibits Cell Detachment Induced by Pemphigus Vulgaris Autoantibodies in HaCaT Cells.

Pemphigus is an autoimmune disease that affects the skin and mucous membranes, induced by the deposition of pemphigus IgG, which mainly targets desmogleins 1 and 3 (Dsg1 and 3). This autoantibody causes steric interference between Dsg1 and 3 and the loss of cell adhesion, producing acantholysis. This molecule and its cellular effects are clinically reflected as intraepidermal blistering. Pemphigus vulgaris-IgG (PV-IgG) binding involves p38MAPK-signaling-dependent caspase-3 activation. The present work assessed the in vitro effect of PV-IgG on the adherence of HaCaT cells dependent on caspase-3. PV-IgG induced cell detachment and apoptotic changes, as demonstrated by annexin fluorescent assays. The effect of caspase-3 induced by PV-IgG was suppressed in cells pre-treated with caspase-3-shRNA, and normal IgG (N-IgG) as a control had no relevant effects on the aforementioned parameters. The results demonstrated that shRNA reduces caspase-3 expression, as measured via qRT-PCR and via Western blot and immunofluorescence, and increases cell adhesion. In conclusion, shRNA prevented in vitro cell detachment and the late effects of apoptosis induced by PV-IgG on HaCaT cells, furthering our understanding of the molecular role of caspase-3 cell adhesion dependence in pemphigus disease.

Effective Electrical Properties and Fault Diagnosis of Insulating Oil Using the 2D Cell Method and NSGA-II Genetic Algorithm.

In this paper, an experimental analysis of the quality of electrical insulating oils is performed using a combination of dielectric loss and capacitance measurement tests. The transformer oil corresponds to a fresh oil sample. The paper follows the ASTM D 924-15 standard (standard test method for dissipation factor and relative permittivity of electrical insulating liquids). Effective electrical parameters, including the tan δ of the oil, were obtained in this non-destructive test. Subsequently, a numerical method is proposed to accurately determine the effective electrical resistivity, σ, and effective electrical permittivity, ε, of an insulating mineral oil from the data obtained in the experimental analysis. These two parameters are not obtained in the ASTM standard. We used the cell method and the multi-objective non-dominated sorting in genetic algorithm II (NSGA-II) for this purpose. In this paper, a new numerical tool to accurately obtain the effective electrical parameters of transformer insulating oils is therefore provided for fault detection and diagnosis. The results show improved accuracy compared to the existing analytical equations. In addition, as the experimental data are collected in a high-voltage domain, wireless sensors are used to measure, transmit, and monitor the electrical and thermal quantities.

CAR-T Cell Therapy: From the Shop to Cancer Therapy.

Cancer is a worldwide health problem. Nevertheless, new technologies in the immunotherapy field have emerged. Chimeric antigen receptor (CAR) technology is a novel biological form to treat cancer; CAR-T cell genetic engineering has positively revolutionized cancer immunotherapy. In this paper, we review the latest developments in CAR-T in cancer treatment. We present the structure of the different generations and variants of CAR-T cells including TRUCK (T cells redirected for universal cytokine killing. We explain the approaches of the CAR-T cells manufactured ex vivo and in vivo. Moreover, we describe the limitations and areas of opportunity for this immunotherapy and the current challenges of treating hematological and solid cancer using CAR-T technology as well as its constraints and engineering approaches. We summarize other immune cells that have been using CAR technology, such as natural killer (NK), macrophages (M), and dendritic cells (DC). We conclude that CAR-T cells have the potential to treat not only cancer but other chronic diseases.

Deregulated cellular circuits driving immunoglobulins and complement consumption associate with the severity of COVID-19 patients.

SARS-CoV-2 infection causes an abrupt response by the host immune system, which is largely responsible for the outcome of COVID-19. We investigated whether the specific immune responses in the peripheral blood of 276 patients were associated with the severity and progression of COVID-19. At admission, dramatic lymphopenia of T, B, and NK cells is associated with severity. Conversely, the proportion of B cells, plasmablasts, circulating follicular helper T cells (cTfh) and CD56- CD16+ NK-cells increased. Regarding humoral immunity, levels of IgM, IgA, and IgG were unaffected, but when degrees of severity were considered, IgG was lower in severe patients. Compared to healthy donors, complement C3 and C4 protein levels were higher in mild and moderate, but not in severe patients, while the activation peptide of C5 (C5a) increased from the admission in every patient, regardless of their severity. Moreover, total IgG, the IgG1 and IgG3 isotypes, and C4 decreased from day 0 to day 10 in patients who were hospitalized for more than two weeks, but not in patients who were discharged earlier. Our study provides important clues to understand the immune response observed in COVID-19 patients, associating severity with an imbalanced humoral response, and identifying new targets for therapeutic intervention.

IL-6 serum levels predict severity and response to tocilizumab in COVID-19: An observational study.

BACKGROUND: Patients with coronavirus disaese 2019 (COVID-19) can develop a cytokine release syndrome that eventually leads to acute respiratory distress syndrome requiring invasive mechanical ventilation (IMV). Because IL-6 is a relevant cytokine in acute respiratory distress syndrome, the blockade of its receptor with tocilizumab (TCZ) could reduce mortality and/or morbidity in severe COVID-19. OBJECTIVE: We sought to determine whether baseline IL-6 serum levels can predict the need for IMV and the response to TCZ. METHODS: A retrospective observational study was performed in hospitalized patients diagnosed with COVID-19. Clinical information and laboratory findings, including IL-6 levels, were collected approximately 3 and 9 days after admission to be matched with preadministration and postadministration of TCZ. Multivariable logistic and linear regressions and survival analysis were performed depending on outcomes: need for IMV, evolution of arterial oxygen tension/fraction of inspired oxygen ratio, or mortality. RESULTS: One hundred forty-six patients were studied, predominantly males (66%); median age was 63 years. Forty-four patients (30%) required IMV, and 58 patients (40%) received treatment with TCZ. IL-6 levels greater than 30 pg/mL was the best predictor for IMV (odds ratio, 7.1; P < .001). Early administration of TCZ was associated with improvement in oxygenation (arterial oxygen tension/fraction of inspired oxygen ratio) in patients with high IL-6 (P = .048). Patients with high IL-6 not treated with TCZ showed high mortality (hazard ratio, 4.6; P = .003), as well as those with low IL-6 treated with TCZ (hazard ratio, 3.6; P = .016). No relevant serious adverse events were observed in TCZ-treated patients. CONCLUSIONS: Baseline IL-6 greater than 30 pg/mL predicts IMV requirement in patients with COVID-19 and contributes to establish an adequate indication for TCZ administration.

Characterization of Dielectric Oil with a Low-Cost CMOS Imaging Sensor and a New Electric Permittivity Matrix Using the 3D Cell Method.

In this paper, a new method for characterizing the dielectric breakdown voltage of dielectric oils is presented, based on the IEC 60156 international standard. In this standard, the effective value of the dielectric breakdown voltage is obtained, but information is not provided on the distribution of Kelvin forces an instant before the dynamic behavior of the arc begins or the state of the gases that are produced an instant after the moment of appearance of the electric arc in the oil. In this paper, the behavior of the oil before and after the appearance of the electric arc is characterized by combining a low-cost CMOS imaging sensor and a new matrix of electrical permittivity associated with the dielectric oil, using the 3D cell method. In this way, we also predict the electric field before and after the electric rupture. The error compared to the finite element method is less than 0.36%. In addition, a new method is proposed to measure the kinematic viscosity of dielectric oils. Using a low-cost imaging sensor, the distribution of bubbles is measured, together with their diameters and their rates of ascent after the electric arc occurs. This method is verified using ASTM standards and data provided by the oil manufacturer. The results of these tests can be used to prevent incipient failures and evaluate preventive maintenance processes such as transformer oil replacement or recovery.

Thermal Analysis of a Magnetic Brake Using Infrared Techniques and 3D Cell Method with a New Convective Constitutive Matrix.

In this work we analyse the temperature distribution in a conductor disk in transitory regime. The disk is in motion in a stationary magnetic field generated by a permanent magnet and so, the electric currents induced inside it generate heat. The system acts as a magnetic brake and is analysed using infrared sensor techniques. In addition, for the simulation and analysis of the magnetic brake, a new thermal convective matrix for the 3D Cell Method (CM) is proposed. The results of the simulation have been verified by comparing the numerical results with those obtained by the Finite Element Method (FEM) and with experimental data obtained by infrared technology. The difference between the experimental results obtained by infrared sensors and those obtained in the simulations is less than 0.0459%.

New Constitutive Matrix in the 3D Cell Method to Obtain a Lorentz Electric Field in a Magnetic Brake.

In this work, we have obtained a new constitutive matrix to calculate the induced Lorentz electric current of in a conductive disk in movement within a magnetic field using the cell method in 3D. This disk and a permanent magnet act as a magnetic brake. The results obtained are compared with those obtained with the finite element method (FEM) using the computer applications Getdp and femm. The error observed is less than 0.1173%. Likewise, a second verification has been made in the laboratory using Hall sensors to measure the magnetic field in the proximity of the magnetic brake.

A single-day point-prevalence study of faecal carriers in long-term care hospitals in Madrid (Spain) depicts a complex clonal and polyclonal dissemination of carbapenemase-producing Enterobacteriaceae.

OBJECTIVES: The objective of this study was to describe the prevalence and microbiological characteristics of carbapenemase-producing Enterobacteriaceae (CPE) colonizing patients in long-term care hospitals (LTCHs) in Madrid, Spain. METHODS: Three LTCHs were included in a single-day point-prevalence survey (September 2013). Rectal swabs, collected from all hospitalized patients (137 in LTCH-A, 121 in LTCH-B and 83 in LTCH-C), were plated onto chromogenic media. Population structure (PFGE and MLST), genes encoding carbapenemases and ESBLs and plasmids carrying carbapenemase genes were characterized. RESULTS: The prevalence of CPE carriers was 4.1% (14/341) [2.9% (4/137), LTCH-A; 4.1% (5/121), LTCH-B; and 6.0% (5/83), LTCH-C]. OXA-48 was the most prevalent carbapenemase (nine Klebsiella pneumoniae, two Escherichia coli, one Enterobacter cloacae and one Citrobacter braakii) followed by VIM-1 (one K. pneumoniae and one Raoultella ornithinolytica). One patient (LTCH-C) was co-colonized with OXA-48-producing K. pneumoniae and E. coli. K. pneumoniae and E. coli isolates also coproduced CTX-M-15 (n = 11) or CTX-M-9 (n = 1) enzymes. K. pneumoniae clustered into six PFGE types corresponding to ST11 (n = 1), ST15 (n = 6), ST307 (n = 1) and ST405 (n = 2). E. coli from LTCH-A and LTCH-C exhibited two different PFGE types associated with ST68. OXA-48 and VIM-1 enzymes were found in different clones in LTCH-A and LTCH-C. However, OXA-48 was the only carbapenemase detected in LTCH-B, mainly associated with K. pneumoniae ST15. KPC, IMP and NDM enzymes were not detected. blaOXA-48 was located on an ∼ 60 kb plasmid with a pOXA-48a-IncL/M backbone. CONCLUSIONS: We describe the first point-prevalence study of CPE faecal carriers in LTCHs in Spain. OXA-48, the most prevalent carbapenemase, showed a complex dissemination pattern with clonal and polyclonal bacterial populations.

Impact of Mean Cell Hemoglobin on Hb A1c-Defined Glycemia Status.

BACKGROUND: Several hematological alterations are associated with altered hemoglobin A1c (Hb A1c). However, there have been no reports of their influence on the rates of exceeding standard Hb A1c thresholds by patients for whom Hb A1c determination is requested in clinical practice. METHODS: The initial data set included the first profiles (complete blood counts, Hb A1c, fasting glucose, and renal and hepatic parameters) of all adult patients for whom such a profile was requested between 2008 and 2013 inclusive. After appropriate exclusions, 21844 patients remained in the study. Linear and logistic regression models were adjusted for demographic, hematological, and biochemical variables excluded from the predictors. RESULTS: Mean corpuscular hemoglobin (MCH) and mean corpuscular volume (MCV) correlated negatively with Hb A1c. Fasting glucose, MCH, and age emerged as predictors of Hb A1c in a stepwise regression that discarded sex, hemoglobin, MCV, mean corpuscular hemoglobin concentration (MCHC), serum creatinine, and liver disease. Mean Hb A1c in MCH interdecile intervals fell from 6.8% (51 mmol/mol) in the lowest (≤27.5 pg) to 6.0% (43 mmol/mol) in the highest (>32.5 pg), with similar results for MCV. After adjustment for fasting glucose and other correlates of Hb A1c, a 1 pg increase in MCH reduced the odds of Hb A1c-defined dysglycemia, diabetes and poor glycemia control by 10%-14%. CONCLUSIONS: For at least 25% of patients, low or high MCH or MCV levels are associated with increased risk of an erroneous Hb A1c-based identification of glycemia status. Although causality has not been demonstrated, these parameters should be taken into account in interpreting Hb A1c levels in clinical practice.

Nano-Regulation of Gene Expression in Chlamydomonas reinhardtii: Harnessing AuNPs for Remotely Switchable Lipid Biosynthesis via Antisense Oligonucleotides.

Antisense oligonucleotide (ASO)-mediated gene silencing has broad applications, spanning from biomedicine to agriculture, involving molecular biology, synthetic biology, and genetic manipulation. This research harnessed nanotechnology to augment ASO-mediated gene silencing, introducing a remotely switchable gene expression system for precise temporal control. We targeted lipid biosynthesis and accumulation enhancement in the photosynthetic eukaryote Chlamydomonas reinhardtii. Gold nanoparticles (AuNPs) transported double-stranded DNA (dsDNA), forming dsDNA-AuNP complexes. These complexes comprised 3'-thiolated sense strands attached to AuNPs and fluorescent antisense oligonucleotides. To avoid harmful laser effects on cells, we adopted a light-emitting diode (LED). Confocal microscopy confirmed dsDNA-AuNP internalization in C. reinhardtii. LED-triggered antisense release led to an 83% decrease in Citrate Synthase 2 (CIS 2) expression. Thiolated sense strand attachment postillumination inhibited antisense reannealing, enhancing gene silencing. This led to significant lipid body accumulation in cells, verified through fluorometric and fluorescence microscopy. This union of nanotechnology and ASO-mediated silencing provides gene regulation opportunities across sectors like biomedicine and agriculture. The system's remote switching capability underscores its potential in synthetic biology and genetic engineering. Our findings substantiate the utility of this approach for enhancing lipid biosynthesis in C. reinhardtii but also underscores its broader applicability to other organisms, fostering the development of novel solutions for pressing global challenges in energy, agriculture, and healthcare.

Baseline Characteristics of Fabry Disease "Amenable" Migalastat Patients in Argentinian Cohort.

Fabry disease (FD) is a multisystem lysosomal storage disorder induced by genetic variants in the alpha-galactosidase A (αGalA) gene. Some FD patients have GLA variants with a reduction in overall αGalA enzymatic activity due to mutated proteins with reduced stability, caused by protein misfolding and premature degradation, but the αGalA catalytic activity remains conserved ("amenable" genetic variants). To correct this misfolding and to prevent premature degradation, migalastat, a small iminosugar molecule was developed. We report the clinical characteristics of FD "amenable" cohort patients from Argentina, prior to starting treatment with migalastat. Seventeen Fabry adult patients were recruited from 13 Argentinian Centers; 8 males (47.1%) and 9 females (52.9%) were included. All genotypes included were missense-type "amenables" mutations. Some classic FD typical early manifestations were more frequent in patients with "classic" versus "late-onset" FD phenotype (pain, p=0.002; cornea verticillata, p=0.019). There was a statistically significant difference in estimated glomerular filtration rate in the "classic" versus "late-onset" phenotype (p=0.026) but no difference between genders (p=0.695). Left ventricular mass was similar between genders (p=0.145) and phenotypes (p=0.303). Cardiovascular risk factors were present among "late-onset" females (obesity 50% and smoke 25%). In patients who started "de novo" migalastat, the main indications were (i) heart disease, (ii) kidney damage, and (iii) pain, while in "switched from prior enzyme replacement therapy" patients, the most frequent indication was "patient decision;" this coincides with publications by other authors.

Machine Learning and External Validation of the IDENTIFY Risk Calculator for Patients with Haematuria Referred to Secondary Care for Suspected Urinary Tract Cancer.

BACKGROUND: The IDENTIFY study developed a model to predict urinary tract cancer using patient characteristics from a large multicentre, international cohort of patients referred with haematuria. In addition to calculating an individual's cancer risk, it proposes thresholds to stratify them into very-low-risk (<1%), low-risk (1-<5%), intermediate-risk (5-<20%), and high-risk (≥20%) groups. OBJECTIVE: To externally validate the IDENTIFY haematuria risk calculator and compare traditional regression with machine learning algorithms. DESIGN, SETTING, AND PARTICIPANTS: Prospective data were collected on patients referred to secondary care with new haematuria. Data were collected for patient variables included in the IDENTIFY risk calculator, cancer outcome, and TNM staging. Machine learning methods were used to evaluate whether better models than those developed with traditional regression methods existed. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: The area under the receiver operating characteristic curve (AUC) for the detection of urinary tract cancer, calibration coefficient, calibration in the large (CITL), and Brier score were determined. RESULTS AND LIMITATIONS: There were 3582 patients in the validation cohort. The development and validation cohorts were well matched. The AUC of the IDENTIFY risk calculator on the validation cohort was 0.78. This improved to 0.80 on a subanalysis of urothelial cancer prevalent countries alone, with a calibration slope of 1.04, CITL of 0.24, and Brier score of 0.14. The best machine learning model was Random Forest, which achieved an AUC of 0.76 on the validation cohort. There were no cancers stratified to the very-low-risk group in the validation cohort. Most cancers were stratified to the intermediate- and high-risk groups, with more aggressive cancers in higher-risk groups. CONCLUSIONS: The IDENTIFY risk calculator performed well at predicting cancer in patients referred with haematuria on external validation. This tool can be used by urologists to better counsel patients on their cancer risks, to prioritise diagnostic resources on appropriate patients, and to avoid unnecessary invasive procedures in those with a very low risk of cancer. PATIENT SUMMARY: We previously developed a calculator that predicts patients' risk of cancer when they have blood in their urine, based on their personal characteristics. We have validated this risk calculator, by testing it on a separate group of patients to ensure that it works as expected. Most patients found to have cancer tended to be in the higher-risk groups and had more aggressive types of cancer with a higher risk. This tool can be used by clinicians to fast-track high-risk patients based on the calculator and investigate them more thoroughly.

Evaluation of food intake through residual analysis in 90 Basque school canteens.

OBJECTIVE: To measure the consumption of 25 food items (Comstock scale), to test whether school management type (public schools vs. private with subsidy schools) and food origin (on-site preparation vs. catering) affected food consumption and to quantify variability associated with territory, school and individual. METHOD: This cross-sectional study involved 14,717 schoolchildren of ages 2-16, recruited in 90 schools of the Basque Autonomous Community (Spain). Waste (non-consumed food) of 25 analysed food items was visually estimated by trained school monitors, via the Comstock categorical scale thus adapted: 1=0%; 2=25%; 3=50%, 4=75% and 5=100%. To analyse the data, mixed modelling was applied. RESULTS: Vegetables, fish and fruits were the less-accepted food types. While school management did not affect food intake, on-site food preparation had better acceptance for legumes with vegetables, oily and lean fish, meat and pre-cooked meals. The largest source of variability in food intake not accounted for by school management and food preparation was the individual subject, while school and territory had moderate and almost no effects, respectively. CONCLUSIONS: Acceptance of the 25 evaluated foods is adequate, albeit can be improved. We believe that promoting on-site food preparation should improve the acceptance of legumes with vegetables, oily/lean fish, meat and pre-cooked meals. We recommend that future interventions oriented to improve intake should focus on individual subjects.

Curcumin Sensitizes 4T1 Murine Breast Cancer Cells to Cisplatin Through PAR4 Secretion.

BACKGROUND/AIM: Prostate apoptosis response 4 (PAR4), a tumour-suppressor protein, selectively induces apoptosis of cancer cells without affecting normal cells. Its soluble form is induced by secretagogues (e.g., chloroquine), and it induces apoptosis by interacting with the receptor of glucose-regulated protein 78, which is overexpressed in cancer cells. In this study, curcumin was analyzed as an inducer of PAR4 expression in 4T1 murine breast cancer cell. and its ability to induce PAR4 secretion in Balb/c mice. In addition, the cisplatin sensitizing effect of soluble PAR4 was analyzed. MATERIAL AND METHODS: The 4T1 cell line was treated in vitro using different concentrations of curcumin; cell viability was analyzed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and PAR4 expression by western blotting. The expression of soluble PAR4 in the serum of mice treated with intraperitoneal curcumin was analyzed using the dot-blot method. Moreover, MTT assay was used to analyze the effects of serum from curcumin-treated mice on cell viability. Tumor size was analyzed in mice treated with curcumin alone and in combination with cisplatin. RESULTS: Curcumin showed a dose- and time-dependent effects on cell viability on 4T1 cells, as well as increasing PAR4 expression. Compared with the control group (phosphate-buffered saline), mice treated with curcumin showed an increase in plasma PAR4. In the Balb/C tumor model, mice treated with curcumin and cisplatin showed greater tumor shrinkage than the control group. CONCLUSION: These results indicate that curcumin induces expression of soluble PAR4 and sensitizes tumor cells to cisplatin.

Mouse mammary tumor virus-like gene sequences are present in lung patient specimens.

BACKGROUND: Previous studies have reported on the presence of Murine Mammary Tumor Virus (MMTV)-like gene sequences in human cancer tissue specimens. Here, we search for MMTV-like gene sequences in lung diseases including carcinomas specimens from a Mexican population. This study was based on our previous study reporting that the INER51 lung cancer cell line, from a pleural effusion of a Mexican patient, contains MMTV-like env gene sequences. RESULTS: The MMTV-like env gene sequences have been detected in three out of 18 specimens studied, by PCR using a specific set of MMTV-like primers. The three identified MMTV-like gene sequences, which were assigned as INER6, HZ101, and HZ14, were 99%, 98%, and 97% homologous, respectively, as compared to GenBank sequence accession number AY161347. The INER6 and HZ-101 samples were isolated from lung cancer specimens, and the HZ-14 was isolated from an acute inflammatory lung infiltrate sample. Two of the env sequences exhibited disruption of the reading frame due to mutations. CONCLUSION: In summary, we identified the presence of MMTV-like gene sequences in 2 out of 11 (18%) of the lung carcinomas and 1 out of 7 (14%) of acute inflamatory lung infiltrate specimens studied of a Mexican Population.